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ISSN 2549-6980

Advances in Tropical Biodiversity and Environmental Sciences Advances in Tropical Biodiversity and Environmental Sciences is a peer-reviewed journal which devoted to the advancement and dissemination of scientific knowledge concerning tropical biodiversity and environmental sciences throughout the world for researchers and professionals. The scope of journal is wide and multidisciplinary that publishes original research papers, review articles, as well as conceptual, technical and methodological papers on all aspects includes research findings, experimental design, analysis and recent application in tropical biodiversity and environmental science studies. This journal published in English and being distributed worldwide. It covers scientific and technological aspects from all fields that have general relevance to tropical biodiversity and environmental sciences including investigations on tropical biodiversity, systematics and taxonomy, terrestrial and aquatic ecology, wildlife management and control, ethnobotany and ethnozoology, tropical plant and animals cultivation, natural product chemistry, ecotourism, environmental remediation and management, and geographic information system (GIS), remote sensing and other modeling application for environmental studies.

ISSN 2549-6980

Advances in Tropical Biodiversity and Environmental Sciences Vol. 1, No. 1, February 2017

Table of Contents Application of Dosage Combinations of Evagrow Biofertilizer and Chemical Fertilizer on Soil Characteristics, Growth and Yield of Rice I Nyoman Merit, I Wayan Narka, and Tatiek Kusmawati

01-05

The Potency of Endofit Fungi in Cocoa as Biological Agent to Control Cocoa Pod Disease Caused by Phytophthota Palmivora (Butler) Butler I Made Sudarma, Ni Made Puspawati, and I Ketut Suada

06-11

Increase Banana Production with Various Applications of Organic Fertilizers I Nyoman Sunarta and Ni Made Trigunasih

12-14

Utilization of Betel Leaf Extract as Botanical Pesticides to Control meloidogyne spp. and Tomato Plant Production Made Sritamin and I Dewa Putu Singarsa

15-17

Marine Biota and Biodiversity: A Sustainable Tourism Perspective Ni Ketut Supasti Dharmawan and Made Sarjana

18-22

Population Control of Viruses Insect Vectors in Chili with Plastic Mulch K.A. Yuliadhi, T.A. Phabiola, and K. Siadi

23-28

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Application of Dosage Combinations of Evagrow Biofertilizer and Chemical Fertilizer on Soil Characteristics, Growth and Yield of Rice I Nyoman Merit*, I Wayan Narka, and Tatiek Kusmawati Program Study of Agroecotechnology, Faculty of Agriculture, Udayana University Bukit Jimbaran, Badung, Bali 80362 *Corresponding author: [email protected]

Abstract. Research on the effect of dosage combination of Evagrow bio-fertilizer and chemical fertilizers on soil properties, growth and yield of rice has been carried out in the glasshouse, Faculty of Agriculture, Udayana University using factorial experiment with a Randomized Block Design (RBD). The first factor is Evagrow bio-fertilizer in 3 level, i.e. E0 (without bio-fertilizers), E1 (5 g bio-fertilizer/L) E2 (10 grams bio-fertilizer/L). The second factor is chemical fertilizer, which consists of 3 levels: K0 (without chemical fertilizers), K1 (150 kg Urea/ha + 75 kg SP36/ha + 37.5 kg KCl/ha), K2 (300 kg Urea/ha + 150 kg SP36/ha + 75 kg KCl/ha). The results showed that chemical fertilizers give a significant and very significant effect on most of the rice growth and yield parameters. The application of chemical fertilizers K1 and K2 increased yield of dry grain harvest to 52.87% and 102.54% compared to controls. Application of Evagrow bio-fertilizer did not significantly increase growth and yield of rice. There is no interaction between chemical fertilizers and Evagrow biofertilizer. Similarly to some of the soil characteristics, biological fertilizer and chemical fertilizer application did not show significant effect on most of the soil characteristics, except on salt levels.

Keywords: Evagrow Biofertilizer, Inorganic Fertilizer, Oryza Sativa

I

INTRODUCTION

Rice (Oryza sativa L.) is the main source of carbohydrate around the world including Indonesia [1]. Demand on rice increase continuously of about 2.23 % per year [2]. Demand of rice increase continuously as population increased, however, this is not followed by increased rice production. Demand for rice reached 32 million ton while current national rice production was only 31.5 ton/ha [3]. Effort to increase rice production via technology development must be done to support food security in Indonesia. Research on application of organic fertilizer such as cow manure, compost, worm manure and green manure has been done. Use of organic fertilizer alone, cannot increase productivity and maintain food security. Therefore, holistic nutrition approach which combine application of organic fertilizer and an-organic fertilizer to increase productivity and environmen-tal sustainability need to be done [4]. Research on the effect of organic fertilizer and an-organic fertilizer on rice growth and yield has been published at Agrivigor Journal, Hasanudin University, Makassar [5]. The use of bio-fertilizer is still limited. Research on the use of Nitrobine bio-fertilizer combine with compost and chemical fertilizer has been done by El-Nagar (2010) on flower plants for 2 seasons [6]. Results of their research shows that optimal dose was 15 ton compost/ha, 3g NPK inorganic fertilizer/pot/month on treatment employ-ing Nitrobine bio-fertilizer 10 g/pot shows the best respond. It revealed that Nitrobine bio-

fertilizer containing Azoto-bacter, Azospirillum and phosphate solving bacteria plays an important role in providing nutrition. II RESEARCH METHOD Research was conducted at a green house, Faculty of Agriculture, Udayana University. Each pot contain 10 kg soil, keep watered for a week to make it muddy and then rice seedlings were planted. Research was conducted in factorial design using Randomized Completely Block Design. There are two factors were examined: Evagrow bio-fertilizer and chemical fertilizer. Evagrow biofertilizer treatment consists of 3 level: without Evagrow bio-fertilizer (EO), 5 g Evagrow bio-fertilizer (E1) and 10 g Evagrow bio-fertilizer per liter (E2). Chemical fertilizer consists of 3 level, i.e: without chemical fertilizer (K0), 150 kg Urea/ha +75 kg SP36/ha + 37,5 kg KCl/ha (K1), 300 kg Urea/ha +150 kg SP36/ha + 75 kg KCl/ha (K2). In total there are nine treatment combinations. Each combination consists of 3 replicates so that there are 27 trial pots. Parameter observed include soil and plants aspect. Soil parameters were nitrogen level (N), phosphorous (P) and Potassium (K), soil pH, C-organic and soil salt level. Growth parameters include: number of shoots, plant height, rice yield parameter include: productive shoots, dry grain weight at harvest, oven dried grain weight, oven dried of dry shoot weight, and oven dried root weight. Nitrogen level (N), phosphorous (P) and Potassium (K), were analyzed using Bray 1 method, C-organic using

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 Walkey & Black method and soil salt level using electric conductometer. Materials needed in this experiment were rice paddy soil sample, rice seedlings, chemicals for soil analyses, Evagrow bio-fertilizer, and chemical fertilizer such as Urea (45% N), SP36 (36% P 2O5), KCl (60% K2O). Equipment required were bucket for growing rice, soil screener, sprayer, oven, digital balance, soil analyses equipment. III

RESULTS AND ANALYSIS

Based on variables observed on rice growth, it can be seen that chemical fertilizer gave significant and highly significat effect on plat height, number and dry weight of shoots. Observation on number of shoot at 56 DAP (days after planting) shows that highest number of shoots were on treatment K2 (23.89 shoots), followed by K1 (18.56 shoots) and lowest K0 (13.44 shoots) or increased 38.02% and 77.69% to control, respectively (Table 1). Increased of number of shoots, shoot dry weight and shoots height was triggered by applica-tion of Urea (46% N), SP36 (36% P2O5) dan KCl (60% K2O), which increased N, P and K availability. Soil use in this experiment has low fertility; it N-total level was 0.120% (low), available-P was 1.77 ppm (very low), available-K was 57.28 ppm (very low). Application of Urea, SP36 and KCl fertiliser has increased N, P and K availability so that increased rice paddy growth. Rice paddy shoots is an important indicator for rice paddy growth. Dry shoot weight was increased on treatment K1 (47.38%) and K2

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(91.27%). Improvement on growth, particularly on number of shoots will increase number of productive shoots (Table 2). Average productive shoots number after application of chemical fertilizer was found highest of treatment K2 (25.67%), which was significantly different with K1 (20.44%) and K0 (14.78%). Increased on number of productive shoots affected fruit weight per shoots. Dry seed weight increased 52.87% and 102.54% on treatment K1 and K2. This increased was caused by improved plant growth, particularly on seedling number. Estimation of grain yield per hectare, with planting space 30 cm x 30 cm was 5.92 ton/ha on K1 and 7.84 ton/ha on K2, while for control was 3.97 ton/ha. This increase was due to fertilizer application which give impact to soil nutrition availability. Increased in nutrition availability improved rice paddy growth, more seedling growth and more productive shoots (Table 2), and resulting in increase on harvested dry grain yield per hectare (Fig. 1). Statistical analyses show that chemical fertilizer and Evagrow bio-fertilizer and its interaction do not give significant impact on shoot/root. Average shoot/root on Evagrow bio-fertilizer treatment was found on E1 (6.69), decreased on E2 (6.00) and lowest on E (5.75), while average shoot/root on chemical application was highest on K1 (6.42) which was not significanty different with K0 (6.01 and K2 (6.00) (Table 2).

TABLE 1. EFFECT OF EVAGROW BIOFERTILISER AND CHEMICAL FERTILIZER ON RICE PADDY GROWTH. Maximum vegetative plant Number of vegetative Oven dried shoot Oven dried root height shoots weight (g) weight (g) (cm) (maximum) E0 87.11 a 18.56 a 39.37 a 14.55 a E1 85.56 a 18.22 a 40.03 a 12.34 a E2 86.89 a 19.11 a 40.38 a 14.29 a 5% LSD K0 82.1 a 13.44 a 27.31 a 9.18 a K1 87.8 b 18.56 b 40.24 b 13.15 a K2 89.7 b 23.89 c 52.23 c 18.84 b 5% LSD 3.30 2,96 4,98 4,35 E0K0 82.3 ab 13.67 a 27.76 a 9.07 a E0K1 89.0 d 18.00 bc 40.54 b 14.26 abc E0K2 90.0 d 24.00 d 49.82 c 20.31 c E1K0 80.0 ab 11.67 a 25.09 a 9.21 a E1K1 87.7 cd 19.67 c 40.85 b 11.40 ab E1K2 89.0 d 23.33 d 54.14 c 16.40 bc E2K0 84.0 abc 15.00 ab 29.07 a 9.27 a E2K1 86.7 bcd 18.00 bc 39.34 b 13.79 abc E2K2 90.0 d 24.33 d 52.73 c 19.82 c Duncan MDRS MDRS MDRS MDRS Note: Numbers that followed by same letter in the same column means it not significantly different on 5% LSD and Duncan 5%. Treatment /parameter

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Fig. 1. Estimation of seed yield per hectare on after fertilizer treatment.

Evagrow application did not show significant impact on growth, yield and soil characteristics. This maybe caused by microbe that contained in Evagrow fertilizer was not active and do not grow well. Hight temperature on the glass house may cause less optimal growth of the soil microbia. This was shown on the average of rice

paddy plants at 7 DAP to 56 DAP did not show significant effect, between E0, E1 and E2, rice paddy height was almost the same.

TABLE 2. EFFECT OF EVAGROW BIO-FERTILIER AND CHEMICAL FERTILIZER ON RICE YIELD PARAMETER. Treatmet Number of Grain weight at Oven-dried grain Estimated dry grain weight Shoot/root /parameter productive shoots harvest (g) weight (g) at harvest/ha (ton) E0 20.33 a 52.57 a 40.11 a 5.84 a 5.75 a E1 20.11 a 51.24 a 38.83 a 5.69 a 6.69 a E2 20.44 a 54.79 a 42.09 a 6.09 a 6.00 a 5% LSD K0 14.78 a 34.82 a 26.43 a 3.87 a 6.01 a K1 20.44 b 53.24 b 40.69 b 5.92 b 6.42 a K2 25.67 c 70.53 c 53.92 c 7.84 c 6.00 a 5% LSD 2,40 5,95 4,49 0,66 E0K0 16.00 a 35.8 a 27.50 a 3.98 a 6.07 a E0K1 19.67 b 52.7 b 40.11 b 5.86 b 5.93 a E0K2 25.33 c 69.2 c 52.72 c 7.68 c 5.26 a E1K0 13.33 a 32.3 a 24.24 a 3.59 a 5.75 a E1K1 21.67 b 53.6 b 40.92 b 5.96 b 7.33 a E1K2 25.33 c 67.8 c 51.33 c 7.53 c 6.99 a E2K0 15.00 a 36.3 a 27.53 a 4.04 a 6.22 a E2K1 20.00 b 53.4 b 41.03 b 5.93 b 6.01 a E2K2 26.33 c 74.7 c 57.69 c 8.29 c 5.76 a Duncan MDRS MDRS MDRS MDRS Note: Numbers that followed by same letter in the same column means it not significantly different on 5% LSD and Duncan 5%.

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TABLE 3. EFFECT OF EVAGROW BIO-FERTILIZER AND CHEMICAL FERTILIZER ON A NUMBER OF SOIL PROPERTIES. Treatment N-total Available P Available K C-organic Soil level (mmhos Soil /parameter (%) (ppm) (ppm) (%) /cm) pH E0 0,23 a 3,86 a 88,18 a 3,51 a 0,72 a 6,94 a E1 0,24 a 3,90 a 88,09 a 3,34 a 0,75 a 6,92 a E2 0,24 a 4,09 a 89,82 a 3,27 a 0,73 a 6,97 a 5% LSD K0 0,22 a 3,53 a 87,31 a 3,18 a 0,56 a 6,96 a K1 0,23 a 4,11 a 89,30 a 3,37 a 0,71 a 6,95 a K2 0,26 a 4,22 a 89,48 a 3,58 a 0,93 b 6,93 a 5% LSD 0,21 E0K0 0,21 a 3,47 a 84,39 a 2,99 a 0,56 a 6,95 a E0K1 0,23 a 4,04 a 89,99 a 3,66 a 0,69 ab 6,91 a E0K2 0,25 a 4,08 a 90,16 a 3,90 a 0,90 bc 6,96 a E1K0 0,23 a 3,41 a 86,78 a 3,27 a 0,58 a 6,96 a E1K1 0,23 a 4,09 a 88,19 a 3,31 a 0,74 ab 6,92 a E1K2 0,26 a 4,19 a 89,31 a 3,43 a 0,95 c 6,88 a E2K0 0,21 a 3,70 a 90,77 a 3,27 a 0,56 a 6,95 a E2K1 0,23 a 4,20 a 89,73 a 3,13 a 0,69 ab 7,01 a E2K2 0,28 a 4,39 a 88,97 a 3,41 a 0,93 c 6,95 a Duncan MDRS Note: Numbers followed by same letters in the same column shows non-significant at 5% LSD or 5% DMRT (Duncan Multiple Range Test). The same thing occurred on other growth parameters such as number of shoot and oven dried shoot weight, did not show significant effect (Table 1). Effect of Evagrow bio-fertilier did not show significant effect on rice paddy growth parameter, also on number of reproduc-tive shoots. Average number of productive shoot. Evagrow bio-fertilizer was found highest on E2 (20.33), decreased but not significant on E0 (20.33 and lowest on E1 (20.11). This maybe due to soil microbia content on biofertilizer, did not develop on soil so that cannot increase soil nutrition availability Analyses on a number of soil properties including Ntotal, Available-P, available-K after chemical fertilizer application revealed an increasing tendency but did not statistically significant. This may due to soil samples was collected at the end of the research. Nutrition that has been given may has decreased its availability. This was because Urea, SP36 and KCL applied was in salt form. Average salt level on chemical fertilizer was found highest on K2 treatment (0,93 mmhos/cm), significantly different with K1 (0,71 mmhos/cm) and K0 (0,56 mmhos/cm (Table 3). Combination between natural fertilizer and chemical fertilizer was expected to have significant interaction, because with the increased of nitrogen, phosphorous and potassium availability will give favorable condition for microbial growth. But in this experiment, interaction between Evagrow bio-fertilizer and chemical fertilizer did not happen. This is because microbial on Evagrow biofertilizer could not grow well, although nitrogen, phosphorus and potassium availability has increased.

IV

CONCLUSION

1.

Application of Evagrow biofertilizer did not show significant impact on soil characteris-tics, growth and yield. 2. Application of chemical fertilizer showed significant impact on almost all growth and yield parameter. Application of 150 kg Urea/ha +75 kg SP36/ha +37,5 kg KCl/ha (K1) and 300 kg Urea/ha +150 kg SP36/ha +75 kg KCl/ha (K2) able to increase harvest yield, each 52.87% and 102.54% compared to control. 3. Applicaton of chemical fertiliser did not show significant impac on soil structure, except for salt level. 4. There is no interaction between Evagrow biofertilizer with chemical fertilizer on all parameter observed. Recommendation It is recommended to continue with field research or set up a trial on difference bio-fertilizer to find out the effect of biofertilizer on plant growth and soil structure. ACKNOWLEDGMENT The authors would like to thank LPPM Udayana University for the research grant. REFERENCES [1] Saragih, B. 2001. Keynote Address Ministers of Agriculture Government of Indonesia, 2nd National Workshop On Strengthening The Development And Use Of Hybrid Rice In Indonesia 1:10.

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[2] Arafah and M. P. Sirappa 2003. Kajian penggunaan jerami dan pupuk N, P, dan K pada lahan sawah irigasi. BPTP Sulawesi Selatan. Jurnal Ilmu Tanah dan Lingkungan 4(1):15-24. [3] Darma, M.D.I. 2007. Swasembada Beras Se-buah Impian? Available: http://www.bali post.co.id/balipostceta/2007/9/17/o2.htm. [4] Suriadikarta, D.A. and R.D.M. Simanungkalit. 2006. Pupuk Organik dan Pupuk Hayati, Organik Fertilizer and Biofertili-zer. Bogor: Balai Besar Penelitian dan Pengembangan Sumberdaya Lahan Pertanian, p. 312.

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[5] Arafah. 2005. Pengaruh Pemberian Pupuk Organik dan Anorganik terhadap Pertum-buhan dan Hasil Padi Sawah. Jurnal Agrivigor 4(2). [6] El-Nagar, A.H. 2010. Effect Biofertilizer, organik compost and mineral fertilizers on the growth, flowering and bulbs produc-tion of Narcissus tazetta, Journal Agricul-ture and Environmental Science 9(1).

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The Potency of Endofit Fungi in Cocoa as Biological Agent to Control Cocoa Pod Disease Caused by Phytophthota Palmivora (Butler) Butler I Made Sudarma*, Ni Made Puspawati, and I Ketut Suada Study program of Agroecotechnology, Faculty of Agriculture, Udayana University Jl. PB. Sudirman, Denpasar, Bali *Corresponding author: [email protected]

Abstract. Cocoa pod disease caused by the fungus Phytophthora palmivora has resulted in loss of cacao in Indonesia, especially in Bali. So far the disease control strategy is not fully understood. So there is a need to find an alternative by using endophytic fungi associated with cocoa plant. Endophytic fungi are needed to be explored in all parts of the cocoa plant such as stems, leaves and husks. The prevalence of fungal endophyte was determined by the size of its domination on the surface tissue for protection against pathogen. The exploration of the endophytic fungi benefits is aimed at finding biological agents that could control of pathogenic P. palmivora. The results showed that 15 types of endophyte fungi have been found in the healthy leaves, stem and pod husks, with the prevalence of fungal endophyte originated from healthy leaf Mecelia sterilia (hyphae sterile) around 30%, the endophyte fungi originated from the healthy cocoa stem are Mycelia sterilia, Neurospora spp and Trichoderma spp around 25%. While the endophytic fungi originated from healthy skin fruit is Trichoderma spp. around 35%. The in vitro test results of endophytic fungi antagonistic against P. palmivora indicated that the endophyte fungi originated from the leaf namely Aspergillus spp was obtained at 80 ± 2%, A. niger 90 ± 2%, A. flavus 100%, and Trichoderma spp. 90 ± 1.5%, the endophytic fungus originated from rods namely Neurospora spp. was 95 ± 2%, and Trichoderma spp. was 90 ± 2%. While the endophytic originated from rind namely Neurospora spp . was 95 ± 1.5 % and Trichoderma spp. was 80 ± 2%. The results of in vivo test of antagonistic endophytic fungi against P. palmivora showed that all of endophytic fungi (Aspergillus sp., A. niger, A. flavus, Neurospora sp., and Trichoderma sp.) have a significant effect in suppressing the growth of mycelium P. palmivora.

Keywords:

Endophytic Fungus, Phytophthora Palmivora, Inhibition, Prevalence, Biological Agents

I

INTRODUCTION

Pod rot is important disease in cocoa cultivation in Indonesia recently, and also in some cocoa producer countries [1]. This disease has average about 20-30% per year on damaging cocoa plant all over the world. In some cases as happen in Samoa America, the cocoa did not planted commercially because of this disease [2]. Cocoa was planted around 532.000 ha in Indonesia on the year of 1999. More than 70% cocoa farmers are the farmer in Indonesia. Indonesia is the top third cocoa exporting countries in the world, produce around 335.000 ton per year, with value 294 million US dollars [3]. Indonesia is the biggest archipelago in the world with has 17.000 islands (6000 has inhabitant). Indonesia is tropical country with various climate and humidity from highland to the lowland. Most of the areas are lowland, while the biggest island has a mountain. Tropical climate with rainy season and high humidity in many areas, some Phytophthora diseases caused a significant damage and uncontrollable. Phytophthora spp. causes disease on the agricultural plants, horticultural and industrial in Indonesia. At least 11 species Phytophthora has been reported as they resulted in economically yield loss in Indonesia. Phytophthora palmivora has been identified to

cause an important disease from economical point of view compared to other Phytophthora species in Indonesia. Phytophthora palmivora infected more than 138 plants species, with caused average loss around 2550% on cocoa plant, while P. capsici caused 52% of decreasing yield on pepper plant. This disease infected many plants but the demage still not be able to be counted, direct demage through fruit infection is showed by the black rotten. Recently, a set of fruits from young fruit to mature one is very sensitive to the infection. [3]. Endophyitic fungi which have been isolated from the tissue of healthy give an new hope for biocontrol of cocoa pod disease. The study from Mejia et al. (2008) showed that 40% (21/52) of isolated endophytic fungi were able to control P. palmivora [4]. One of the isolated fungi which has antagonist characteristic through a simple competition mechanism is Trichoderma sp. II

RESEARCH METHOD

Endophyte Isolation Endophytic fungi used in this study were collected from cocoa planted in Tabanan. The survey of cocoa leaf and fruit was done in four different locations at cocoa plantation centre in Tabanan regency. The obtained leaf and fruit were washed by flowing water with following

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 methods: 32 peaces of leaf with size 4 mm2, the shoot and the flower were cut from the middle of each, the surface was sterilized in 0,525% sodium hypochlorite for 3 minute, and 70% ethanol for 2 minute; and washed by sterile water for 1 minute; and then put in the PDA media (containing antibiotic livoploxacyne 0,1% (w/v). The grown fungi from the piece of leaf were transferred to the tube containing PDA for storage and classified through morphospecies. In order to isolate the endophyte from rod and fruit, those parts were washed by flowing water and then devided into 8 pieces. Fruit were cut into 16 cubical form (8 mm3), 8 from exocarp and 8 from mesocarp. The surfaces were sterilized and storage with similar procedure as isolation from the leaf. Identification of Endophytic Fungi The storage endophytic fungi were grown on petri disc containing PDA and repeated 5 times. The cultures were incubated at dark room and temperature ±27oC. The isolate was identified macroscopically after 3 days of culture periode by observe of the colony colour, growth rate, and microscopically identification was done to investigate septa on the hyphae, the spora form/conidia and the sporangiophore. Fungi identification was done by using some reference books of Samson et al., 1981; Pitt and Hocking, 1997; Barnett and Hunter, 1998; Indrawati et al., 1999 [5][6][7][8]. Prevalence of Endophytic Fungi Determination of endopyhtic fungi prevalence were done based on frequency of endophytic fungi isolate were found (eight pieces from leaf, rod, flower and fruit) per petri disc, divided by all founded isolates timed by 100%. The number of isolate prevalence will be determining the dominancy of the endophytic fungi on the healthy cocoa plant. Inhibition Assay of Endophytic Fungi to the Pathogen The ability of each endophytic fungi to inhibit the growth of pathogenic P. palmivora was tested by dual culture technique (one pathogen colony in the middle and two endophytic fungi next to the pathogen, side by side). Their inhibition can be calculated as follow [9][10]: A–B Inhibitory (%) = x 100 A A= B=

colony diameter P. palmivora in the single culture (mm) colony diameter P. palmivora in the dual culture (mm).

In Vivo Antagonistic Assay In vivo antagonistic assay of endophytic fungi was done by impaling the fresh fruit by small spleden needle for 20 times, and then covered by the spore of antagonist fungi (spore from one petri dish was dissolved in 250 ml sterile aquadest), then dyed into the spore of pathogenic fungi (P. palmivora). The assays were setting as follows: A = control (without covering by antagonist) B = Antagonist treatment 1 (spore suspension 5x107) C = Antagonist treatment 2 (spore suspension 5x107) D = Antagonist treatment 3 (spore suspension 5x107)

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E = Antagonist treatment 4 (spore suspension 5x107) F = Antagonist treatment 5 (spore suspension 5x107) All of the treatment was repeated 5 times. Experiment was designed by random group design, and after variance analysis by ANOVA following by signi-ficance test at level of 5%. The infection parameters were measured by counting the number of infected impale divide by all impale (20 times) timed by 100%. III

RESULTS AND ANALYSIS

Endophytic Fungi The results showed that endophytic fungi obtained from cocoa plant namely the endophyte from leaf were 7 isolates of Micelia sterilia with, 2 isolates of Aspergillus spp., 3 isolates of Aspergillus niger, 1 isolate of Aspergillus flavus, 1 isolate of Fusarium sp., 1 isolate of Mucor sp., 5 isolates of Trichoderma spp. and 1 isolate of Verticillium sp. Endophyte from the rod were 5 isolates of Micelia sterilia, 1 isolate of Botryoderma sp. 1 isolate of Dactylium sp. 2 isolates of Fusarium sp., 1 isolate of Oidium sp. 5 isolates of Neurospora spp. and 5 isolates of Trichoderma spp. The endophyte obtained from the skin fruit were 3 isolates of Micelia sterilia, 1 isolate of Cylindrocarpon sp., 2 isolates of Fusarium sp. 1 isolate of Mortierella sp. 5 isolates of Neuro-spora spp., 1 isolate of Septocylindrium sp. and 7 isolates of Trichoderma spp. as showed in Table 1 and Fig. 1. Endophytic fungi normally exist without any symptoms (asymptomatically) in the tissue of the host plant and they have strong attachment to their host. There are two main reasons of thus attachment i.e. first, they are growing indicated that endophyte is able to found in all kind of plant with high abundance and vary. Most of this endophyte are found in internal infection site at leaf, root, rod and skin then transmitted horizontally through the spore. Secondly, the endophyte can produce mycotoxin and convert the physiology and morphology of the host plant. Mycotoxin from endophyte has an advantage to the host plant as acquired plant defenses to face herbivore insect and grass host [11]. Endophyte from leaf and rod of Endofit yang berasal dari daun dan Hevea brasiliensis which frequent to found are the genus of Penicillium, Pestalotiopsis and Trichoderma [12]. Aspergillus and Fusarium produced bioactive compound from the host which contain insecticide activity, cytotoxic and anticancer [13]. The study from Amin et al. (2014) have found 6 genus of endophytic fungi in VSD M.05 resistant cocoa plant, namely Curvularia sp., Fusarium sp., Geotrichum sp., Aspergillus sp., Gliocladium sp., and Colletotrichum sp., and another four were not be able to identified [14]. Prevalence of Endophytic Fungi This study showed that prevalence of endophytic fungi originated from healthy leaf is Mycelia sterilia (sterile hypha) around 30%, in endophyte from healthy cocoa rod are Mycelia sterilia, Neurospora spp. Trichoderma spp. around 25% of each. While in the endophyte from the fruit skin is Trichoderma spp. 35%

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017 (Table 1). Trichoderma spp., are the dominant genus in south part of China (4 isolates, 23,5%). It has been reported that endophytic fungi can produce antitumor or antifungi activity. The fungi which has been isolated from

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the skin fruit of medicinal plant including Pasecilomyces sp., Cephalosporium sp., Mortierella sp., Mucor sp., Trichoderma sp., and Cladosporium sp. [15].

TABLE 1. THE ENDOPHYTIC FUNGI PREVALENCE ON THE LEAF, ROD AND FRUIT OF THE HEALTHY COCOA PLANT. Fungi Micelia sterilia Aspergillus spp. Aspergillus niger Aspergillus flavus Botryoderma sp. Cylindrocarpon sp. Dactylium sp. Fusarium sp. Oidium sp. Mortierella sp. Mucor sp. Neurospora spp Septocylindrium Trichoderma spp. Verticillium sp Total

Endophyte from the leaf 6 (30%)* 2 (10%) 3 (15%) 1 (5%) 1 (5%) 1 (5%) -

Endophyte from the rod 5 (25%) 1 (5%) 1 (5%) 2 (10%) 1 (5%) 5 (25%)

Endophyte from the fruit skin 3 (15%) 1 (5%) 2 (10%)

5 (25%) 5 (25%) 1 (5%) 20 20 *Note: the percentage is prevalence (frequency of isolates)

1 (5%) 5 (25%) 1 (5) 7 (35%) 20

Endophyte from leaf Endophyte from stem Endophyte from fruit peel

Fig. 1. Endophytic fungi originated from leaf, rod and fruit skin of cocoa plant.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 The Results of Antagonistic Assay Endo-phytic Fungi on Phytophthota palmivora The result of this study showed that its obtained around 80±2% of Aspergilus spp., 90±2% of A. niger, 100% of A. flavus and 90±1,5% of Trichoderma spp in endophyte from leaf. 95±2% of Neurospora spp. and 90±2% of Trichoderma spp. are obtained from endophyte which originated from rod. While 95±1,5% of Neurospora spp. and 80±2% of Trichoderma spp. from endophyte originated from the fruit skin (Table 2, Fig. 2).

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Aspergillus flavus and A. terreus which growing internally are seed transmitted fungi that can inhibit the growth of pathogenic Rhizoctonia solani Khun, and then it’s useful for the control of seed transmitted diseases which infected several important plant [16]. Most endophyte which is found in cocoa rod are the genus that familiar known as soil fungi (Clonostachys and Trichoderma) [4].

TABLE 2. INHIBITORY ASSAY OF ENDOPHYTIC FUNGI TO P. PALMIVORA Fungi Micelia sterilia Aspergillus spp. Aspergillus niger Aspergillus flavus Botryoderma sp. Cylindrocarpon sp. Dactylium sp. Fusarium sp. Oidium sp. Mortierella sp. Mucor sp. Neurospora spp Septocylindrium Trichoderma spp. Verticillium sp

Endophyte from leaf 80±2% 90±2% 100% 90±1,5% -

A

D

B

E

Endophyte from rod 95±2% 90±2% -

Endophyte from fruit skin 95±1,5% 80±2% -

C

K

Fig. 2. Antagonistic assay of endophytic fungi to P. palmivora, (A) Aspergillus sp., (B) Aspergillus niger, (C) Aspergillus flavus, (D) Neurospora sp., (E) Trichoderma sp., and (K) Control (Phytophthora palmivora) 7 day after incubation Trichoderma sp. also found as endophtic fungus which is able to inhibit Crinipellis perniciosa (Stahel) Singer, the main cause of Witches Broom disease on

cocoa plant [17]. Trichoderma, has potency as a biological agent to control Phytophthora spp., moreover

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017 Fusarium and Verticillium are antagonistic to some pathogenic fungi [18]. In Vivo Antagonistic Assay The in vivo antagonistic assay showed that all five biological agents have a significant different on inhibition of the P. palmivora growth. The five biological agents that tested on the fresh fruit i.e. Aspergillus sp., A. niger, A. flavus, Neurosporas sp., Trichoderma sp. (Fig. 3).

A

B

A

A

D

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Among endophytic fungi founded on Hevea brasiliensis, Trichoderma is the most often been found and isolated [12], and is able to inhibit the growth of P. palmivora either by competition or antibiosis mechanism. The percentage of infection are significantly different in all treatment compared to the control (Fig. 3, Table 3). Antagonistic treatment using Aspergillus sp. has percentage infection around 10%, while the other (A. niger, A. flavus, Neurospora sp. and Trichoderma sp.) has percentage infection around 5% each.

C

B

D

E

K

Fig. 3. The result of in vivo antagonisticE assay on fresh fruit (A) Aspegillus sp., (B) Aspergillus niger, (C) Aspergillus flavus, (D) Neurospora sp., (E) Trichoderma sp. and (K) Control 7 days after incubation

TABLE 3. THE INFECTION PERCENTAGE OF P. PALMIVORA ON THE FRUIT IN EACH TREATMENT. No. 1 2. 3. 4. 5. 6.

IV

Tretment Percentage of Infection Control 100 ± 0 a ** Treatment with Aspergillus sp. 10 ± 0,2 b Treatment with Aspergillus niger 5 ± 0,1 b Treatment with Aspergillus flavus 5 ± 0,3 b Treatment with Neurosporas sp. 5 ± 0,2 b Treatment with Trichoderma sp. 5 ± 0,1 b ** Significant different at BNT 1%.

CONCLUSION

Endophytic fungi which have been found on the leaf, rod and fruit skin of healthy cocoa are 15 species, with prevalence of endophytic fungi from healthy leaf are Mecelia sterilia (sterile hifa) around 30%, on the endophyte from healthy rod it’s found mycelia sterilia, Neurospora spp. and Trichoderma spp. around 25% of each. While at the endophyte originated from fruit skin, it’s found Trichoderma spp. around 35%. The in vitro antagonistic assay results of endophytic fungi to P.

palmivora showed that at the endophyte from leaf its obtained Aspergilus spp. around 80±2%, A. niger 90±2%, A. flavus 100%, and Trichoderma spp. around 90±1,5%. On the endophyte from rod, it’s obtained 95±2% of Neurospora spp. and 90±2% of Trichoderma spp. While endophyte from the fruit skin, its obtained 95±1,5% of Neurospora spp. and 80±2% of Trichoderma spp. The result of in vivo antagonistic assay showed that all of endophytic fungi (Aspergillus sp., A. niger, A. flavus, Neurospora sp., dan Trichoderma sp.) have a significant effect on inhibition the mycelium growth of P. palmivora.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 ACKNOWLEDGMENT We acknowledge the rector of Udayana University, the head of the LPPM, The dean of Faculty of Agriculture, Udayana University, for all the support from the start until finishing of this research. REFERENCES [1] Semangun, H. 1991. Penyakit-Penyakit Penting Tanaman Perkebunan di Indonesia. Yogyakarta: Gadjah Mada University Press. [2] USDA, 2012. Phytophthora palmivora Pests and Diseases of American Samoa. American Samoa Community College Community & Natural Resources Coope-rative Research & Extension 12:12. [3] Purwantara, A., D. Manohara and J.S. Warokka. 2004. Phytophthora Diseases in Indonesia. In Diversity and Management of Phytophthora in Southeast Asia Edited by A. Drenth and D.I. Guest, ACIAR Monograph 114:70-75. [4] Mejia L.C., E.I. Rojas, Z. Maynard, S.V. Bael, A.E. Arnold, P. Hebbar, G.J. Damuels, N. Robbins, and E.A. Herre. 2008. Endophytic fungi as biocontrol agents of Theobroma cacao pathogens. Biological Control 46:4-14. [5] Samson, R.A., E.S. Hoekstra, and C.A.N. Van Oorschot. 1981. Introduction to Food-Borne Fungi. Centraalbureau Voor-Schimmelcultures. Institute of The Royal Netherlands. Academic of Arts and Sciences. [6] Pitt, J.I. and A.D. Hocking. 1997. Fungi and Food Spoilage. 2nd Edition. London: Blackie Avademic and Professional. [7] Barnett, H.L. and B.B. Hunter. 1998. Illustrated Genera of Imperfect Fungi. APS Press. Minnesota: The American Phytopathological Society. [8] Indrawati. G., R.A. Samson, K. Van den TweelVermeulen, A. Oetari and I. Santoso. 1999. Pengenalan Kapang Tropik Umum. Yayasan Obor Indonesia. University of Indonsia Culture Collection Depok, Indonsia and Centraal bureau voor Schirmmelcultures, Baarn, The Netherlands. [9] Dolar, F.S. 2001. Antagonistic effect of Aspergillus melleus Yukawa on soilborne pathogens of Chickpea, Tarim Bilimleri Dergisi 8(2):167-170. [10] Mojica-Marin, V., H.A. Luna-Olvera, C.F. Sandoval-Coronado, B. Pereyra-Alférez, H. Lilia,

[11] [12]

[13]

[14]

[15]

[16]

[17]

[18]

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Morales-Ramos, E. Carlos, Hernández-Luna, and G.O. Alvarado-Gomez. 2008. Antago-nistic activity of selected strains of Bacillus thuringiensis against Rhizoctonia solani of chili pepper. African Journal of Biotechnology, 7 (9):1271-1276. Faeth, S.H. 2002. Are endophytic fungi defensive plant multualist? Oikos 98:25-36. Gazis, R. and P. Chaverri. 2009. Diversity of fungal endophytes in leaves and stems of wild rubber trees (Hevea brasiliensis) in Peru. Fungal Ecology 3:240254. Zhao, J., L. Zhou, J. Wang, T. Shan, L. Zhoung, X. Liu, and X. Gao. 2010. Endophytic fungi for producing bioactive compound originally from their host plants. In Current Research, Technology and Educa-tion Topics in Applied Microbiology and Microbial Biotechnology. A. Mendez-Villas (Ed.) Formatex pp.567- 576 Amin, N., M. Salam, M. Junaid, Asman and M. S. Baco. 2014. Isolation and identification of endophytic fungi from cacao plant resistant VSD M.05 and cacaomplant susceptible VSD M.01 in South Sulawesi, Indonesia. International Journal of Current Micorbiology and Applied Science. 3(2):459-467. Saithong, P., W. Panthavee, S. Stonsaovapak, and L. Congfa. 2010. Isolation and primary identification of endophytic fungi from Cephalotaxus manii trees. Maejo Int. J. Sci. Technol. 4(03): 446-453. Sharma, A.K., P. Sharma and R.B. Sharma. 2013. Characterization of anti-fungal property of seed coat leafchates of Jatropa curcas L. IJBAF 1(10):446451. Rubini, M.R., R.T. Silva-Ribeiro, A.W.V. Pemella, C.S. Maki, W.A. Araujo, D.R. dos Santos, and J.L. Azevedo. 2005. Diversity of endophytic fungal community of cacao (Theobroma cacao L.) and biological control of Crinipellis perniciosa, causal agents of witches broom disease. Int. J. Biol. Sci. 1:24-33. Mpika, J., B.I. Kebe and F.K. N’Guessan. 2011. Isolation and Identification of Indigenous Microorganisms of Cocoa Farms in Côte d’Ivoire and Assessment of Their Antagonistic Effects VisÀ-Vis Phytophthora palmivora, the Causal Agent of the Black Pod Disease, Biodiversity Loss in a Changing Planet 303-318.

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Increase Banana Production with Various Applications of Organic Fertilizers I Nyoman Sunarta* and Ni Made Trigunasih

Staff and Lecturer of Agrotechnology Department, Faculty of Agriculture, Udayana University, Bukit Jimbaran, Bali, Indonesia. *Corresponding author: [email protected]

Abstract. Banana is a fruit that must be present in every activity of the Balinese people. Almost in every cultural and religious activity it is compulsory to present the banana fruit, even more Bali as a tourism area really need fruits including bananas. Therefore the need of bananas is highly demanded in traditional markets or supermarkets in Bali. Until now, the banana production in Bali is far below demand, therefore we still import form places outside of Bali. Banana has a high nutrient value due to nutrients found in it such as potassium and folic acid that is needed by the body. It is also a good source of calcium, phosphor, nitrogen, and vitamins such as vitamin A, vitamin C, and B complexes which helps to repair and regenerate tissues of the body. Banana plants can grow in many places, from low grounds until highlands and in various types of soil. Nevertheless for the optimal growth; fertile soil, crumbly thick, lots of humus, aeration, and a good drainage as well as enough water is all needed. By optimizing land use, banana plants are often planted as a sideline plant for plantation of coconut, cocoa, and coffee, as well as sideline plants for many other in-between plantations. The way of planting the banana is a conventional method that is commonly used by local farmers including those in the Angkah village. For growth of banana plants we must consider and ensure before planting a crumbly soil when it is solid, to make drainage, and to make levels in slopes. During plantation organic/compost fertilizers is needed as much as 15-20 kg for each hole for plantation. Organic fertilizers that are added influences the good production for both quantity and quality which makes the fruit tastier and has a higher nutritional value. There are some types of organic fertilizers that are used in this research that are: cow waste organic fertilizer, chicken waste organic fertilizer, pig waste organic fertilizer, goat waste organic fertilizer, and compost fertilizer with a dosage of 15 kg per plant. The placement of fertilizers for each places of treatment is conducted by using a Random Group Design (RGD). The total treatments are 6 and are repeated 3 times. Planting is conducted on the 7th of July 2016 with a ground hole size of 60cmx60cm and a depth of 50 cm. The seedlings that are used are decedents that are 50-65cm in height. The specific aim that is wished to be achieved is the increase production of banana as sustainable food in Indonesia. The result up to the progress of this research was found that treatment with chicken waste fertilizer has given the best influence for growth of the banana plant, increased number of leaves, as well as increased height of the plants, all compared to the controlled. The increment of banana plant with chicken waste fertilizer was 63.33 cm for 3 months.

Keywords: Banana, Nutritional Values, Production, Organic Fertilizer

INTRODUCTION I

INTRODUCTION

Banana is the most popular fruit in the community especially in Bali. Almost in every cultural and religious ceremony there must be some banana fruits, even more Bali as a tourism area needs fruits especially bananas. Until today the production of banana in Bali is far from what is needed, therefore it is imported from places outside of Bali such as the island of Java, Lombok, Sumbawa, and even from Celebes. Banana fruit has a high nutritional value because of its nutritious composition such as potassium and folic acid which is highly needed during pregnancy. It is also a good source of calcium, phosphor, nitrogen, and vitamins such as

vitamin A, vitamin C, and B complexes which helps to repair and regenerate tissues of the body. Due to its good nutritional values, therefore banana is a very good healthy food as a crop fulfillment. Banana plants can grow in many places, from low grounds until highlands and in various types of soil. Nevertheless for the optimal growth; fertile soil, crumbly thick, lots of humus, aeration, and a good drainage as well as enough water is all needed. By optimizing land use, banana plants are often planted as a sideline plant for plantation of coconut, cocoa, and coffee, as well as sideline plants for many other in-between plantations. The way of planting the banana is a conventional method that is commonly used by local farmers including those in the Angkah village.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 A good media for plantation will be very helpful for the growth of the bananas. As stated by Anon. (2016) of which to achieve a good growth of banana plants, therefore before plantation it is necessary to ensure a crumbly soil when it is solid hard, to make drainage, and to make levels in slopes [1]. During plantation organic/compost fertilizers is needed as much as 15-20 kg for each ground hole for plantation [2]. Organic fertilizers that are added influences on a good production for both quantity and quality which makes the fruit tastier and has a higher nutritional value [3]. There are a few types of organic fertilizers found in the Angkah village that can be used such as: cow waste organic fertilizer, chicken waste organic fertilizer, pig waste organic fertilizer, goat organic waste fertilizer, and compost fertilizer from plant and other organic wastes. The utilization of organic fertilizers are not yet optimal, it is shown by much manure are scattered in the farmer’s fields (near the animal housings) which may be drifted away during rainy season, especially for the pig waste. Organic fertilizers are very useful for plants because it could provide macro and micro nutrients, the media for root growth will be better, also it has the same characteristic as a soil such as; soil are more crumbly, can restrain moist longer, better soil aeration and drainage, and can reduce the drifting of soil during raining seasons [4]. Considering the reasons above therefore it is needed to conduct a research/evaluation of the use of various types of organic fertilizers that are found for banana plants so it could be useful to use manure waste and more sustaining a better environment. On the other hand, it is hoped that the production of the bananas could increase; therefore the fulfillment of fruits as food stocks can be optimal. By the increased production, it is hoped that farmers could earn more; therefore the prosperity of farmers can be elevated. II

RESEARCH METHOD

The materials used in this research are seedlings of Green Thailand Bananas as much as 50 plants, cow waste organic fertilizer, chicken waste organic fertilizer, pig waste organic fertilizer, goat waste organic fertilizer, and compost fertilizer. The tools that are used in this research are plastic bags for soil and manure sampling, stationaries for observation and measurement in the field, a hoe, a sickle, knife/cutter, and other things. This Research is a field experiment which uses a factorial design with a fundamental Random Group Design (RGD). This research is commenced by achieving soil samples to analyze the nutrients. Soil sample that are taken in different pieces are achieved by the depths up to 30 cm in some of the research area and are then combined into one soil sample. The soil sample is dried by wind and then analyzed in the laboratory to find out the results of: pH

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(H2O), N-Total, P2O5 availability, K-Total, salinity, and organic materials in the soil, as well as the texture of the soil. For organic fertilizer that is used, samples are also taken to analyze the composition values of: pH (H 2O), N total, P2O5, and K2O. Experiment with the organic waste fertilizers are treated by the followings: 1. P0 : Without fertilizers (controls) 2. PS : Treatment with cow waste organic fertilizer at 15 kg per plant 3. PA : Treatment with chicken waste organic fertilizer at 15 kg per plant 4. PB : Treatment with pig waste organic fertilizer at 15 kg per plant 5. PK : Treatment with goat waste organic fertilizer at 15 kg per plant 6. Kom : Treatment with organic compost fertilizer at 15 kg per plant All of the six treatments are each repeated 3 times, giving a total of 18 treatment areas. Observation of the research is started by looking at the vegetative development of the plant such as: height of the plant, amount and width of the leaves, flower timing (banana buds), and the weight of the fresh fruit. Results of the observation are analyzed statistically with differential Duncan test. III

RESULTS AND ANALYSIS

The research results of influences of the types of organic fertilizers towards the growth and production of banana plantation are up until this paper was made has just been up to the observation aspect of the plantation. Observation of the vegetative growth was the increment of the amount of leaves and the height of the plant. The analysis result of the organic fertilizers and the soil of the research location can be found in Table 1. On the table above shows that the composition or nutrients and organic soil in the research area were very low, therefore it is very much needed to be fertilized, especially with an organic fertilizer. The average growth of the plant can be seen in the observation result of the amount of leaves and the height of the banana plants found in Table 2 and Table 3. Observation results of the amount of leaves and height of the banana plant was found that chicken waste organic fertilizer had the highest influences compared to the other organic fertilizer. There is a real influence of organic waste fertilizer towards the height of the banana plantation. This is caused by the composition of nutrient in the soil is low (table 1), and the composition of nitrogen, phosphate, and potassium elements in chicken waste fertilizer is the most highest compared to the other organic fertilizer. With the increase height of the plant and the amount if banana leaves is hoped to produced fruits which are much more abundance and faster.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017

No. 1 2 3 4 5 6

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TABLE 1. ANALYSIS RESULT OF ORGANIC FERTILIZER AND THE SOIL OF THE RESEARCH GROUNDS Result of Analysis Item that are analyzed pH (1:2,5) C Organic (mmhos/cm) N total (%) P2O5 (ppm) K2O (ppm) Texture Cow waste 7,1 N 28,85 ST 0,63 T 533,86 ST 550,80 ST Pig waste 7,2 N 20,27 ST 2,78 ST 410,27 ST 520,54 ST Chicken waste 7,4 N 24,04 ST 3,27 ST 977,61 ST 1074,35 ST Goat waste 7,2 N 23,80 ST 2,16 ST 816,53 ST 446,36 ST Compost 6,8 N 17,39 ST 0,53 T 706,67 ST 589,51 ST Soil of research ground 6,7 N 1,72 R 0,18 (R) 0,27 (SR) 71,07 (SR) Thick clay Notes: N= Neutral; R = Low; SR = Very Low; T = High; ST = Very High

TABLE 2. INFLUENCES OF TREATMENTS TOWARDS AMOUNT OF BANANA LEAVES Total Leaves (Sheets) On The Age Of The Plant Treatment 1 Month 2 Month 3 Month 4 Month P0. Controlled 3.67 6 7.33 8,00 a PB. Pig waste 4.33 6.67 7.67 7,67 a PA. Chicken waste 5.67 7.33 8.67 8,67 a PK. Goat Waste 5.33 7.33 8.67 8,67 a PS. Cow Waste 5.00 7 7.67 8,67 a Kom. Compost 4.67 6.67 7.67 8,33 a Note: The numbers that are followed by the letters in the same column shows there are differences with no real difference during the BNT test at 5%.

TABLE 3. INFLUENCES OF TREATMENT TOWARDS THE HEIGHT OF THE BANANA PLANTS Height of Banana Plant at Different Ages Treatment During plantation (cm) 1 Month (cm) 2 Month (cm) 3 Month (cm) 4 Month (cm) P0. Controlled 55.67 57.67 61.33 71.35 89,33 a PB. Pig waste 51.33 53.33 57.33 84.67 103,33 d PA. Chicken waste 53.33 57 66.67 92.33 116,33 e PK. Goat Waste 62 64.33 68.00 77.33 99,67 c PS. Cow Waste 61.67 64.33 68.33 76 98,00 c Kom. Compost 50.67 55.67 59.00 72.67 93,67 b Note: The numbers that are followed by the letters in the same column shows there are differences with no real difference during the BNT test at 5%.

IV CONCLUSION

REFERENCES

The soil in the research location really needs to be fertilized especially with organic fertilizers. The chicken waste organic fertilizer has the best influences and was significant towards the growth (height) of the banana plant which was 115,3% from the time it was planted. While the controlled plant only grown by 88,33% by the time of plantation.

[1] Anonymous. 2016. Budidaya Pisang. Available from: http://www.produknaturalnusantara [2] Bellamy, A. S. 2013. Banana Production Systems: Identification of Alterna-tive Systems for More Sustainable Production. Ambio. 42(3):334-343. [3] Soeparjono, S. 2016. The Effect of Media Composition and Organic Fertilizer Concentration on the Growth and Yield of Red Ginger Rhizome (Zingiber officinale Rosc.). Agri-culture and Agricultural Science Procedia Vol. 9:450-455. [4] Ssali, H., B. McIntyre, and C. Gold. 2003. Nutrient Cycling in Agroeco-systems 65:141. doi:10.1023/A: 1022184927506.

ACKNOWLEDGMENT The researcher would like to thank the Dean of Agriculture Faculty and Head of LPPM of Udayana niversity for the help that is given so this service of research can be conducted. This research and services is funded by DIPA PNBP of Udayana University based on the legal letter For Research commencement Number: 1268/UN14.1.23 /PL/2016, dated on the 21 st of September 2016.

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Utilization of Betel Leaf Extract as Botanical Pesticides to Control meloidogyne spp. and Tomato Plant Production Made Sritamin* and I Dewa Putu Singarsa Agroechotechnology Program, Faculty of Agriculture of Udayana University *Corresponding author: [email protected]

Abstract. Meloidogyne spp. Are among the polyphagous pest that has spread around the world and has been reported to attack cultivated plants with economic values. Various ways had been applied to control the Meloidogyne spp. nematodes but has not yet showed any effective results. We are currently using synthetic pesticides because of its fast response and ability to maintain plant productions. The improper application of synthetic nematicides would have a negative impact on the environment. The aim of this research is to know the effectiveness of betel leaf concentration (Piper betel L.) to curb the development of nematodes on Root-Knot Meloidogyne spp and the growth of tomato plants. This is a descriptive research, with the utilization of betel leaf extract (Piper betel L,) by concentration treatment of 0 %, 5 %, 10 %, 15 % and 20%, each of which are repeated five times. Complete Random Sampling (CRS) with the F test is used and continued with the Duncan test at a 5 % rate. The result shows that the betel leaf extract of the 20% concentration was the most effective either on the growth of plants and to the root-knot nematodes. The suppression of growth in the ground was 80,06% and the lowest was recorded with the 5% concentration ( 2,32 %); the suppression of root-knot was 45,45 % while the lowest was with the 5% concentration (12,12 %); the suppression of nematodes population in the roots was 45,45 % while the lowest was with the 5% concentration (39,76 %); and last was the suppression of egg mass which was 61,73 % while the lowest was shown in the 5% concentration (18,32 %). It can be concluded in this research that the most effective betel leaf extract concentration was at 20%.

Keywords: Meloidogyne spp., Piper betel L., CRS

I

INTRODUCTION

There was a decrease in tomato plant productions from 647.020 tons in 2005 to 629.744 tons in 2006 [1]. One of the important pests causing the decrease of tomato production is the root-knot nematodes causing root ulcers, Meloidogyne spp. These nematodes have an important role in causing damage on the roots of horticultural plants, crops, plantation, and weeds [2]. The damage caused by Melodogyne spp. especially in tomato plants around the world is significant. Meloidogyne spp. are among the concerning pests due its polyphagous characteristic and its growth population has spread around the world [3]. The Meloidogyne spp. nematode attacks almost every vegetable plant and some plants can be attacked by more than one nematodes species. Meloidogyne spp. are spread all around the world and many have been reported to attack cultivation plants with economic values, serious losses could occur when plants are severely infested. Agrios (1969) stated that losses due to root-knot nematodes Meloidogyne spp. are variable depending on the type of plant being infested, the species of Meloidogyne, and the environment condition [4]. If

young susceptible plants are infested, it would cause it to die, however if an adult plant are infested, it would have only a small effect towards the production. From previous studies using many plant leaves extract as botanical pesticides, it is found that the use of betel leaf extract was the best way to suppress nematodes population, however it is not yet known the most effective concentration of its botanical use to control nematodes [5]. Meloidogyne spp. nematodes attack almost all vegetable plants and some of those plants can be attacked by more than one nematode species. Meloidogyne spp. are spread all across the world and have been reported to attacked many cultivation plants with economic values, of which severe losses can happen if the plants are severely infested. Agrios (1969) stated that the loss due to root-knot nematodes Meloidogyne spp. varies depending on the types of plants. Various ways of controlling is applied towards rootknot nematodes Meloidogyne spp. which includes plantation of nematode resistant varieties, plant rotation, and technical culture, however these controlling methods is less effective to suppress Meloidogyne spp. population [6]. Until now, many farmers are still using synthetic pesticide on tomato plantation to control nematodes due

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 to its fast response and ability to maintain plant productions; however improper application of synthetic nematicides will have a negative outcome towards the environment. The purpose of this research is to know the effectiveness of the concentration level of betel leaves extract to suppress the development of root-knot nematodes Meloidogyne spp. in tomato plants, to know the development of tomato plants after being treated with betel leaves extract and also to know the result of tomato plant productions. The major purpose in this research is to support farming and to increase their family income, especially for tomato productions. II

RESEARCH METHOD

The research is conducted in an experiment garden of the Agriculture Faculty in conjunction with Laboratory of Pests and Plant Disease of Agriculture Faculty Udayana University, of which is a descriptive research which utilize piper betel leaf extract as treatment with the concentration of 0%, 5%, 15% and 20%. Each treatment is done with five repetitions which are given to tomato plants which have been infested with root-knot nematodes, Meloidogyne spp. This research uses a Complete Random Design (CRD) analyzed by F-test and continued with Duncan’N 0,05 (5%). III

RESULTS AND ANALYSIS

The research result shows that all treatment that was given had a real effect towards all changes that was observed. Towards the growth of plants, the concentration of betel leaf extract that was used, has given different effects on the changes observed, one of which the 20% concentration has given the best result towards plant growth compared to the other, while the growth is retarded with the usage of lower concentration. This result is supported by other studies conducted by

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Ambika and Poonima (2014) in India which states that treatments with Kirinyuh leaf extract applied into the soil of soy bean plantation can increase the height of the plant by 15 %, root length by 40%, and attached beans by 163%. Statistical analysis result showed real differences between treatment with 20% concentration compare with the controls and between treatments which had shown real differences presented in Table 1. IV

CONCLUSION

As for the nematodes population in the soil, the 20% concentration has had the biggest suppression at 80,06%, and the smallest was with the 5% concentration with 2,32%; as for the amount of root-knot, the 20% concentration suppression was 45,45% and lowest was with the 5% concentration at 12,12%; as for nematodes in the roots, the 20% concentration suppression was 76,14% and lowest was with the 5% concentration at 39,76%; and for the amount of egg mass with the 20% concentration suppression was 61,73% and lowest was with the 5% concentration at 18,32%. In this research, it has been concluded that the most effective concentration of betel leaf extract is 20%. Treatment applied for plant growth showed the best plant growth also occurred with the 20% concentration treatment while the least growth occurred with the 5% concentration treatment. From the result it can be concluded that the best concentration of betel leaf extract is with the 30% concentration. Feedback: 1. Need to conduct further research in the field with the effective concentration (20%) with tomato plantation. 2. Needs to conduct further research with other types of plants and different types of nematodes; on different types of plants that are attacked by root-knot nematodes or parasitic nematodes

TABLE 1. THE INFLUENCE OF BETEL LEAF EXTRACT APPLICATION IN VARIOUS CONCENTRATIONS TOWARDS SOME CHANGES.

Note: Numbers of which are followed by different letters in the same columns has a result of significantly different at the level of testing with Duncan’N 5% (0,05)

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017 17 [2] Dropkin, VH. 1991. Pengantar Nematologi ACKNOWLEDGMENT Tumbuhan Edisi 2. (Terjemahan). Yogyakarta: Gadjah Mada University Press. 1. To the Headmaster of Udayana University who had [3] Adiputra, M. G. 2006. Pengantar Nematologi given us the chance to conduct this research until Tumbuhan. Jurusan Hama dan Penyakit Tumbuhan completion. Fakultas Pertanian Universitas Udayana. 2. To the head of LPPM of Udayana University who [4] Agrios, G.N. 1970. Plant Pathology. 2nd Printing. had given me the chance to conduct this research New York: Academic Press. 629 p. through the budget listing of Budget conduction [5] Sritamin, M., I N. Wijaya, dan I D.P. Singarsa. program of Udayana University. 2015. Efektifitas Berba-gai Konsentrasi Ekstrak 3. To the team of research and university students who Jenis Daun Tanaman Terhadap Populasi Nematoda had been involved in this research. Puru Akar, Meloidogyne spp. dan Hasil Tanaman Tomat. Laporan Hasil Penelitian Hibah Unggulan REFERENCES Udayana 2015. [1] Badan Pusat Statistik dan Direktorat Jendral Bumi [6] Kerry, B.R. 2001. Exploitation of the nematophagus dan Produksi. 2006. Produksi Tomat Menurut Fungal Verticullum chlamydosporum Godard of the Provinsi Tahun 2002-2006. Available: Biological Control of Root-Knot Nematodes. http://www.bps.go.id//produksitomat/ [15 June 2013].

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Marine Biota and Biodiversity: A Sustainable Tourism Perspective Ni Ketut Supasti Dharmawan1 and Made Sarjana2

1

Civil Law Department, Faculty of Law, University of Udayana, Jalan Bali No.1, Denpasar, Bali, 80114, Ph./Fax: 0361 222666, Email: [email protected] 2 Civil Law Department, Faculty of Law, University of Udayana, Jalan Bali No.1, Denpasar, Bali, 80114, Ph./Fax: 0361 222666, Email: [email protected]

Abstract. Protection and conservation of marine biodiversity and their utilization based on sustainable environment, balance, and fairness play an important role in the context of sustainable tourism. The importance of protecting the availability of a healthy environment, which not only focuses on the present, but also a sustainable environment for future generations, is regulated under several provisions such as: Law of the Republic of Indonesia Number 27 Year 2007 on the Management of Coastal Areas and Small Islands, Law No. 32 of 2009 on the Environmental Protection and Management, Law No. 10 Year 2009 on Tourism, Law No. 5 Year 1990 on the Conservation of Biological Diversity and Ecosystems, the UN WTO Global Code of Ethics for Tourism, and the Convention on Biological Diversity. In order to protect and conserve marine biodiversity, it is considered relevant to use deep ecology approach in realizing the presence of marine biota and sustainable biodiversity for the entire ecosystem of life, including human life ecosystems in the development of tourism activities. States and all stakeholders have responsibility for the protection and conservation of biodiversity, including sustainable development of marine biota and its diversity for tourism activities.

Keywords: Biodiversity, Protection, Conservation, Responsibility, Sustainable Tourism

I

INTRODUCTION

Article 5 letter b of Law No. 10 Year 2009 on Tourism (Tourism Law) regulates linkages between tourism activities with Human Rights, especially based on the principle of individual right which one of the embodiment is the right to tourism [1]. However, such Tourism Law also regulates the rights aimed at the interests of the Third Generation of Human Rights, namely its collective nature or intended for the public. That can be seen in the Considering section letter (a) which determines that the nature, flora and ancient relics, relics of history, art and culture of the Indonesian nation constitute as resources and capitals of tourism development to increase the prosperity and welfare of the people as contained in Pancasila and the Preamble of the 1945 Constitution of the Republic of Indonesia. The resources and capitals of tourism development are very important to be maintained in order to establish a sustainable tourism. The Third Generation of Human Rights is also known as the Collective Rights or Peoples Rights [2]. Vasak (1990) suggested that the Third Generation of Human Rights is based on the principles of fraternity or solidarity while the First and Second

Generation of Human Rights are based on the principle of Individual Rights [3]. The fulfilment of the individual rights, such as the right to tourism supposes not making the rights of society as a whole to be reduced or neglected. In order to fulfil the right of everyone to tourism, various tourism activities have been developed. In the end it is realized that tourism activities actually bring in foreign exchange and improve people's welfare and national development. Such development also aims to attract tourists to see the view of various rivers, mountains and the beautiful expanse of nature, to the tourist areas known as the underwater attractions. The underwater attractions present the beauty inside the sea. By diving for example, reefs and wide variety of species of fish and marine life will be seen and enjoyed by tourists. Indonesia’s underwater tourism is widely known in international level. One of them is located in Tulamben, Karangasem of Bali Province. Tulamben is very popular because it is easy to reach the location, it has beautiful coral reefs and a variety of unique underwater creatures and the diversity of marine fish ranging from small ones like sea slugs, crabs, shrimps, jack fish, lion fish, garden eels, ghost pipe fish and pygmy seahorse until up to the big ones as sharks, molamola. Tulamben

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 also supported by beatiful coral reefs that sticks to the walls of the cargo ship United USAT Liberty Glo sinking, who difers at a depth of 5 to 30 meters can enjoy the cracks ship united with marine life, coral reefs and variety of unique fishs which are sweaming around the venue. Moreover, Tulamben also offers a wide range of products related to the submarine tourism such as: offering many dive sites suitable for diving courses, fun dive and underwater photography. The charm of the underwater flora and fauna as well as the beauty of diversity marine life on one hand can support enjoyment of the tourists. They enjoy, happy, and may more fresh after vacation surrounding the marine tourism areas. However, on the other hand the beauty of the underwater attractions and a variety of marine life and its biodiversity can be potentially endangered as a result of excessive and irresponsible exploitation of tourism activities. The sea with its biodiversity is losing its quality standards and function as a result of such overuse. The tourism sector is also criticized as the sector that is supported by the sea as its charm which in the end leaves another problem related to the sustainability of biodiversity including the ecosystems of underwater biota. In relation to this phenomenon, it is important to examine the responsibility of tourism sector related with protection and conservation to marine biota and its biodiversity. This study focuses on the study of: how is the regulation of marine biodiversity related to sustainable tourism? As well as who is responsible for the management of protection and conservation of the biodiversity? The purpose of this paper is to analyse the regulation of marine biodiversity, including in the national and international dimension as well as to analyse the responsibility of the state and relevant stakeholders in the management of protection and conservation of marine life biodiversity. II

RESEARCH METHOD

This is a normative legal research which employs Statute, Conceptual, Comparative as well as Deep Ecology approaches. The examined legal materials are the primary and secondary legal materials. All legal materials were analysed in descriptive qualitative. III

RESULTS AND ANALYSIS

The Regulation on Marine Biodiversity Conservation in Tourism Activities Point (c) of the Consideration section of Tourism Law stipulates that tourism is an integral part of national development which is done in a systematic, integrated, sustainable and responsible ways while providing protection towards religious values, the living culture in society as well as sustainability and quality of the living environment. In relation to that, the appropriate stakeholders of tourism activities, especially the corporations that take advantage of the panoramic of underwater biota and biological diversity as part of its activities, together with the government and the state are responsible for realizing the tourism activities oriented to

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the sustainable management, conservation and environmental protection. In improving sustainable tourism activities especially those which are supported by the charms of the sea, several provisions can be seen as relevant, namely: Article 5 paragraph (1), Article 20 paragraph (1) and Article 33 paragraph (3) of the 1945 Constitution of the Republic of Indonesia and Law No. 27 Year 2007 on the Management of Coastal Areas and Small Islands [4][5]. Coastal areas and small islands are part of the natural resources given by God Almighty which wealth is controlled by the state and must be preserved as well as its utilizations are for the greatest prosperity of the people, both for the present and future generations. In this regard, the exploitation of marine life and biodiversity in tourism activities shall refer to the Law No. 27 Year 2007. Indonesia is famous as a country with its beautiful and extensive of coral reefs. Indonesian coral reef preservation plays a very important role both nationally, regionally and globally - a total area of the world’s coral reefs reached 284.300 km2. Of the area, the Indonesian water is the one who has the most extensive coral reef, which is 51.020 km2 (18% of the world's coral reefs) followed by Australia (48.000 km2) and the Philippines (25.000 km2). Because of it is extensive and its diversity of species is very high, Indonesia is mentioned by the oceanographers as the centre of global coral triangle. Although Indonesia has beautiful and extensive coral reefs, however it cannot be denied that the reefs in Indonesian water are suffering serious destruction and therefore requires attention from all parties. The prohibition against malicious activity on coral reefs stipulated in Article 35 of the Law No. 27 Year 2007 which basically sets the prohibitions on taking coral reefs in the conservation areas, using explosives, toxic substances, and/or other substances that destroy coral reef ecosystems and taking equipment, means, and other methods that destroy coral reef ecosystems. The provision of Article 1 paragraph (3) of Law No. 32 of 2009 on the Environmental Protection and Management also regulates sustainable development as a conscious and planned effort that combines aspects of environmental, social, and economic development strategies to ensure the environmental integrity and safety, capability, prosperity and quality of life of present and future generations [6]. With regard to the achievement of tourism sustainable development, there are three important pillars that must be considered and maintained in harmony, namely: the balance of economy, environmental and social coupled with the pillar of climate change. The issue of climate change has an influence both directly and indirectly to the development of economy and social culture [7]. The regulation on biodiversity can also be found in Article 2 of the UN Convention on Biological Diversity 1992 (CBD): “Biological diversity” means the variability among living organism from all sources including, inter alia, terrestrial, marine and other aquatic ecosystems and the ecological complexes of which they are part: this includes diversity within species, between species and of ecosystems [8]. Based on the definition set out in Article

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017 2 of the CBD, Kundis Robin Craig (2005) identified three (3) important elements in regulating the definition of biodiversity, namely: genetic diversity, species diversity and ecosystem diversity [9]. Besides that, such Article 2 also stresses that biodiversity including the study of biodiversity in the terrestrial, marine and other aquatic ecosystems and the ecological complexes which they are part (other aquatic ecosystems). Indonesia as one of the countries that ratified the CBD through Law No. 5 Year 1994 stipulates that "biodiversity" is diversity among living organisms from all sources, including, terrestrial, marine and other aquatic ecosystems as well as the ecological complexes that are part of its diversity, this includes diversity within species, between species and of ecosystems. The existence of biodiversity both in the marine, terrestrial and other aquatic ecosystems are very important to be protected and preserved because its existence has a very large contribution to the quality of human life. The United Nations Environmental Program (UNEP), stressed that the loss of biodiversity threatens our food supplier, opportunities for recreation and tourism, as well as sources of wood, medicines and energy [10]. To emphasize the importance of preserving the existence of biodiversity, Indonesia as a member state, in the letter (b) of the Considering section stipulates that The existence and sustainability of biodiversity includes ecosystems, species and genetic includes animals, plants and microorganisms need to be guaranteed for life. Indeed the existence of biodiversity, including marine life is very helpful. One of the benefits is to support food chains and nets. Biodiversity supports ecosystem stability, namely: food chains and nets are in balance, the balance of species composition, as well as the more complex components of the ecosystem, the more efficiently the ecosystem will last [11]. In order to support policies and agreements to the CBD, a number of protocols have been issued in international level such as: the Cartegena Protocol on Biosafety to the Convention on Biological Diversity, and the Nagoya Protocol on Access to genetic Resources and the Fair and Equitable Sharing of Benefits Arising from their Utilization to the Convention on Biological Diversity (Nagoya Protocol) [12]. Regulation on the protection and conservation of biodiversity including marine life is very important to be enforced, especially in the context of sustainable ecosystem. It is because sustainable ecosystems always put balance and harmony, interdependence between humans and the natural surroundings based on the deep ecology with ecosophy approach and its relevance with human rights. The utilization of the current biodiversity resources has relevance to ensure sustainable environment included therein in order sustainable tourism. Article 1 Paragraph (2) and Paragraph (3) of Law No. 5 Year 1990 on the Conservation of Biological Diversity and Ecosystems regulates matter on the management of natural resources which utilization is wisely used to ensure a balance supply while maintaining and improving the quality of its biodiversity and values, as well as the existence of natural resource ecosystems [13]. It is a system of interrelationships between

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elements in nature, both living and non-living, which are mutually dependent and influencing. Protection and conservation of marine biota constitute an integral part of the marine protection in a broader context, as can be seen from the Law No. 32 Year 2014 on Marine (Marine Law). Marine conservation cannot be separated by the activities or efforts towards the sea. Indonesian marine law also emphasized the importance of the underlying management and utilization of marine resources based on the notion of blue economy. Based on the elucidation of Article 14 Paragraph 1 of Marine Law, it is noted that “blue economy” is an approach to improve the sustainable marine management and conservation as well as coastal resources and their ecosystems in order to realize economic growth with the principles among others: community involvement, resource efficiency, waste minimization, and multiple revenue. Gunter Paulli, originator of the first blue economy concept which criticized the weakness of the concept of the green economy, argues that the blue economy is the economic development based on marine aspects, but not only exploit marine resources but also the maintain and protect marine ecosystems. This concept is a form of marine industrialization and fisheries policies which is based on modernization. In other words, this blue economy accelerates economic growth by exploiting the marine and fisheries potential [14]. The blue economy concept is very relevant to be applied in marine management in Indonesia, including in the biodiversity and marine biota protection and conservation. It is because as it is known, Indonesia is the largest archipelago in the world with 95.181 km long of coastline, and with the sea area of 5.4 million km2. Indonesia has huge potential marine resources, including its biodiversity and the largest non-living marine and aquaculture of the world. In connection to Indonesia’s marine potential, the government’s role is needed to develop the marine-based economic sectors. One idea that can be applied to accelerate the development of marine economy of Indonesia is to use the concept of the blue economy (Ibid.). Stakeholder’s Responsibilities in Protecting Marine Biota and Biodiversity The continuation existence of biodiversity and ecosystems, including marine biota as has been stated previously, is very beneficial to the surrounding environments, including human life. The extinction of one species of biodiversity will disturb other ecosystems, therefore, in a variety of regulations that intersect with the environment including marine environment, it is easily to be found provisions on the protection and conservation which are able to guarantee the sustainability of the ecosystem for the benefit of future generation. In other words, sustainable development, sustainable environment and sustainable tourism aimed not only to the present life, but also ensure its survival for future generation. Holistically with the principle of Deep Ecology approach, in order to ensure the sustainability of biodiversity and the whole ecosystem, the frontline responsibility is on the shoulders of the state. The provision on it can be seen

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 through the general elucidation of Law No. 32 Year 2009 which regulates that a good and health living environment constitutes as human rights and constitutional rights of every citizen of Indonesia [15]. Therefore, the state, the government and all stakeholders are obliged to carry out sustainable environmental protection and management therefore environmental still being as source of life and support of the Indonesian people as well as other living things. Article 235 of the United Nations Convention on the Law of the Sea (hereinafter referred to as UNCLOS) of 1982 determines that each state is responsible for fulfilling international obligations concerning the protection and preservation of the marine environment. Each state should ensure the availability of effort in their law system on how to obtain compensation promptly and adequately with regard to the damage caused by natural persons or legal entities under its jurisdiction. Each state should cooperate in implementing the international law governing the responsibility and liability indemnity for compensation for losses due to pollution of the marine environment, as well as the payment procedure. Indonesia is one of the states that have ratified the UNCLOS 1982. Article 25 in conjunction with Article 26 of Marine Law stipulates that in order to utilize and manage the marine industry (one of which industrial biotechnology), the Central Government and the Local Government are responsible to develop and enhance the marine biotechnology industry which is done by utilizing the potential of biodiversity aimed to transform and prevent the extinction of marine life as a result of exploration excess, develop environmentally friendly technology at every marine industry, and develop marine resource management system on an on-going basis. Besides that, all relevent stakeholders in the utilization of biological diversity are responsible together with the government to ensure the protection and implementation of conservation and to prevent environmental pollution so it is not damaging or interfering with the survival natural resources biodiversity including marine biota. Provisions governing the responsibilities of stakeholders can be seen in: Article 23-26 of Tourism Law, as well as Article 3 the UN WTO [16]. IV

CONCLUSION

1. Regulation on the protection and conservation of the biodiversity and marine life is set out in a range of conditions both internationally as the United Nations Convention on Biological Diversity in 1992 and its Protocols or nationally which basically stipulates that biodiversity is including diversity among living organisms from all sources, including, terrestrial, marine and other aquatic ecosystems as well as ecological complexes that are part of its diversity, this includes diversity within species, between species and ecosystems. Conservation activities are very important to be maintained, protected and carried out its presence is very beneficial to the quality of human life, especially in the context of sustainable human life in the human rights dimension, including sustainable tourism.

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2. The parties responsible for the law enforcement in relation to the protection of biodiversity and marine life in the context of sustainable tourism are the state and all stakeholders including corporations in tourism sector associated by various laws such as the CBD, the UN WTO, Law on Environmental Protection and Management, Tourism Law and Marine Law. Suggestion 1. Government is expected to make Action Plan with regard to the protection and conservation of biodiversity, particularly marine biota and disseminate it in a sustainable manner, especially to the relevant stakeholders engaged in tourism activities. Therefore, the existence of biodiversity of marine life will be save and continue to be useful in the context of the fulfilment of human rights towards ecosystem resources for the future generations. 2. Governments, law enforcement officers and academics are expected to disseminate the responsibilities of all stakeholders to jointly enforce the law both in the local, national and international levels with regard to the protection and conservation of biodiversity, in particular, marine biota. ACKNOWLEDGMENT Through this article, we would like to deliver our sincere gratitude to the Dean of the Faculty of Law, University of Udayana, Head Master of Notary and LPPM of University of Udayana for the financial and moral supports therefore this HUPS research and its publication can be held on time. REFERENCES [1] Law No. 10 Year 2009 on Tourism [2] Freedman, R. 2013. Third generation Rights: Is There room for Hybrid Constructs Within International Human rights Law, Cambridge Journal of International and Comparative Law (2)4:935. [3] Bülent. A. 2004. Rethinking Third Generation Human Rights, Ankara Law Review Vol. l, No. 1. Summer 2004. [4] The 1945 Constitution of the Republic of Indonesia. [5] Law No. 27 Year 2007 on the Management of Coastal Areas and Small Islands [6] Law No. 32 of 2009 on the Environmental Protection and Management. [7] Chatarayamontri, N. 2009. Sustainable Tourism And The Law: Coping wih Climate Change (Dissertation & Theses), Paper 6, Available: http://digitalcommons.pace.edu /lawdissertations/6. [8] The UN Convention on Biological Diversity. [9] Craig, R. K. 2005. Protecting International Marine Biodiversity: International Treaties and National Systems of Marine Protected Areas, Journal of Land Use Vol.20, No.2., Spring 2005. [10] UNEP (Secretariat of the Convention on Biological Diversity). 2000. Sustaining life on Earth How the Convention on Biological Diversity Promotes Nature and Human Well-Being.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017 [11] Arthana, I.W. 2016. Tantangan dan Solusi Perlindungan dan Pemanfaatan Berkelanjutan Keanekaragaman Hayati di Bali, makalah dalam Konsultasi Publik Diplomasi Indonesia menuju Konferensi Negara Pihak ke-13 Konvensi PBB Keanekaragaman Hayati, 16 Juni 2016. [12] Arsika, I M.B. 2011. Implikasi Ratifikasi Dan implementasi Atas Konvensi PBB mengenai Keanekaragaman Hayati, Makalah dalam Konsultasi Publik Diplomasi Indonesia menuju Konferensi Pihak ke-13, 16 Juni 2016.

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[13] Law No. 5 Year 1990 on the Conservation of Biological Diversity and Ecosystems. [14] Satria, A. E. 2014. Ekonomi Biru Sebagai Prinsip Ekonomi Berkelanjutan di Indonesia, Available: http://www.kompasiana.com/andikaekasatria/ekono mi-biru-sebagai-prinsip-ekonomi-berkelanjutan-diindonesia_54f4b808745513a32b6c8d3b [15] Law No. 32 Year 2014 on Marine [16] The UN WTO Global Code of Ethics for Tourism.

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Population Control of Viruses Insect Vectors in Chili with Plastic Mulch K.A. Yuliadhi *, T.A. Phabiola and K. Siadi

Faculty of Agriculture, Udayana University, Bali, Indonesia *Corresponding author: [email protected]

Abstract. The incidence of diseases caused by pathogenic viruses in chili is still a major problem in reducing the production of chili in Indonesia. Most agricultural crops are hosts for one or more types of plant viruses, so the virus continued to be a problem in the tropics. Virus is passive, requires intermediaries vector to be transmitted to other plants. The goal of this research was to develop control strategies for aphids that act as a viral vector and pest chili plants using plastic mulch. Control design that was developed in this study based on the habits of local farmers, using plastic mulch with two colors, black and silver. Mulching is done to dispel the arrival aphids into the chili crop, at the same time preventing the emergence of weeds that act as alternative hosts of the virus. The use of silver plastic mulch to control vector viral populations was better compared to black plastic mulch during chili planting. The use of silver plastic mulch can improve yields of chili crops.

Keywords: Aphid, whiteflies, Chili chili, virus

I

INTRODUCTION

The chili production is still low in Indonesia, which merely 4.35to/ha, while the potential production are more than 10ton/ha. One of the cause is plant pests that affect from nursery through the post-harvest. Among the plant pests, that most affecting the production of chili is the group of viral pathogens. Viral infection on chili plant reduced between 32-75% of its production [1] or up to 68.22% found by Nyana (2012) [2]. The host range of viruses that cause disease in chili is very broad, including intermediate host that can provide a source of inoculum at any time, such as some weeds. Weeds that are growing around chili plants can lead to competitions among the and may also be an alternative host of the viruses [3]. Chili plant virus remains a problem to this day. This is because the source of inoculum available throughout the year, abundance of the vector that are active all the time, and there is no insulation between the gardens. Insects are the most important vectors of plant viruses. There were approximately 700 known species of plant viruses in 1991, which 426 species are transmitted by insects [4]. The species of Aphids have been known to avoid silver reflection of light [5]. The avoidance of silver light by the insects gives us an opportunity to use plastic mulch to plant crops. Control of virus spread was aimed of this study by preventing the contact between infective aphids carry viruses (viruliferous) with chili cultivated plants.

Prevention was done by implementing silver and black plastic mulch. II

RESEARCH METHOD

The research has been conducted at Kerta Village, Payangan, Gianyar Regency. The research was begins with the soil tillage and seed preparation, installation of plastic mulch in the beds, planting, maintaining and harvesting crops. This study was dsesigned in a randomized block design with 3 treatments and 9 replications. All three treatments tested were: 1. Planting virus-free seedlings without mulch (K), 2. Planting virusfree seedlings with silver plastic mulch (MP) and 3. Planting virus-free seedlings with Black plastic mulch (MH). Confirmation of viral infection is done in serological testing by ELISA technique. Serology detection with ELISA technique was performed using symptomatic plants mosaic with specific antiserum CMV and TMV, while for ChiVMV was using antiserum Universal Potyvirus with DAS-ELISA method following the procedure described in the kit antiserum used (Agdia, USA). Value absorbance was measured at 405 nm with an ELISA Reader Data was analysed. In order to determine the effect of the treatments given, the observed data was analysed in the analysis of variance (ANOVA), if there was a significant different among treatments (p ≤ 5%) observed, further Duncan analysis was conducted.

Journal of Advances in Tropical Biodiversity and Environmental Sciences, p-ISSN: 2549-6980 III

RESULTS AND ANALYSIS

Apids Population Insect virus vector found in this study was the species of aphids and whiteflies, the species of whitefly was Besimia tabaci (Table 1). The average population of

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aphids and whiteflies changes in accordance with the host plant growth stages of each period of observation. Myzus persicae population at the beginning of the observations increased (Fig. 1), in line with the growth of chili plants and culminated in the observation VI.

Fig. 1. The average abundance of Myzus persicae at time observation (individuals/plant).

An increase in the population of both species of aphids M. persicae and A. gossypii from the initial observation period to the highest in the observation period VI, because the chili or chili plant was in peak period of vegetative growth, and the availability of young leaves attract the M. persicae and A. gossypii to visit. Young leaves provide food sources for aphids. Aphids can grow optimally when plants sprout [6]. The abundance of M. persicae and A. gossypii will decrease when the plant get older or entering the generative period, the plant tissue is tougher and the cell fluid was reduced, so it can support the aphid life. The abundance of aphids I

chili crops was closely related to the plnat metabolism activities [7] and the quantity and quality of plant nutrition. Reduction of food supply will affect the abundance of aphids. The low population of B. tabaci population was observed at the beginning of study in chili plantation. In line with the growth of chili plants, the average population of B. tabaci has increased and reached its peak in the observation VI, then the presence of B. tabaci declined and the lowest when plants at the age of 12 week after planting (Fig. 3).

Fig. 2. The average abundance Aphis gossypii at time observation (individuals/plant).

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Fig. 3. The average Bemisia tabaci at time observation (individuals/plant).

The population of B. tabaci was abundance when chili plants were in vegetative phase, on the observation VI, and decreased in generative phase (algorithmic) on observation XII which may follow the growth of plants.

The quantity of plants can be measured by the number of plant biomass, while the plant quality is depended on various plant nutrition content that are required by insects [8]

TABLE 1. THE AVERAGE ABUNDANCE OF MYZUS PERSICAE, APHIS GOSSYPII AND BEMISIA TABACI ON OBSERVATION VI Treatments M. persicae

Average abundance (idividuals) A. gossypii B. Tabaci

NT 21.8 a 22.1 a 18.4 a MH 18.9 b 15.2 b 13.2 b MP 11.8 c 10.9 c 8.3 c Note: The number followed by different letter in similar column significantly different at the Duncan test at 5%. NT: no treatment/control; MH (black plastic mulch); MP (silve plastic mulch) Statistical analysis showed that the average population of vector species either M. persicae, A. gossypii and B. tabaci in silver mulch treatment was significantly different from the population of each vector in black mulch treatment and control. The low average population of each vector of disease-causing viruses on chili plants in the silver mulch treatment because silver has the ability to reflect about 33 percent of near ultra violet light [9], light waves that are favored by most insects, so the insects will follow the direction of reflection and leaving crops [10]. Light reflection is able to reduce the heating effect of rhizosphere under the plastic surface, and the range of light favored by insects, so the insects will follow the direction of reflection and leaving crops. Consequently insect populations act as vectors of diseasecausing viruses can be reduced in the planting area [11]. Virus Disease Symptoms The results showed that chili plants in controls (NT) showed symptoms of the virus, wich were higher compared to the treatment of black plastic mulch (MH) and silver plastic mulch (MP), with the symthom of mosaic viruses (61.7%), yellow (21%) and chlorosis

(9.1%) at the observation of 10 week after planting (Table 2). The average crop with the mosaic virus symtoms was highest in the control treatment, as many as 61 plants was affected. Based on ELISA test results, it was found that CMV-infected plants is the ultimate for all treatments (Table 3). Based on the result of ELISA test (Table 3), there were several types of viruses associated with mosaic disease in chili ie. CMV, TMV, and ChiVMV. The highest symtoms of virus attack was found in control plant treatments, followed by chili plants treated in black plastic mu;ch and the lowest was in silver plastic mulch (Table 3). Similar trend was found by Nyana (2012), that the chili with mosaic symptoms were associated with three different virus types, namely Tobacco mosaic virus (TMV), Cucumber mosaic virus (CMV) or Chilean veinal motle virus (ChiVMV).

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TABLE 2. THE PERSENTAGE OF CHILI PLANTS WITH THE VIRUS SYMTOMS IN EVERY TREATMENT (WITHOUT MULCH/CONTROL, BALCK PLASTIC MULCH AND SILVER PLASTIC MULCH). The percentage of plant with virus symtoms (week after planting) 6 8 10 M K Kl M K Kl M K NT 32.9 8.7 6.8 45.6 12.0 8.6 61.7 21.0 MH 9.0 5.4 3.8 14.0 8.0 4.8 23.0 14.0 MP 5.0 2.9 1.5 7.7 3.3 2.3 10.1 4.8 Note: NT: control, MH: black plastic mulch, MP: silver plastic mulch, M: mosaic, K: Yellow, Kl: Chlorosis

Treatments

Kl 9.1 6.8 3.4

TABLE 3. THE PERCENTAGE OF VIRUS-INFECTED PLANTS WITH MOSAIC SYMPTOMS OF DAS-ELISA TEST RESULTS OF THE PLANTS ON THE OBSERVATION 10 WEEK AFTER PLANTING Number of symptomatic mosaic plant Mosaic

CMV

TMV

ChiVMV

NT MH

61 23

22 9

19 4

19 8

MP

10

3

1

2

Treatments

Number of virus infected plant*

*The present of the virus base on DAS-ELISA test.

The high percentage of plants symptomatic CMV in each treatment (control, black and silver plastic mulch) is due to this type of virus has a wide range of plant hosts, including the weeds as an intermediate host, so provide source of inoculum all the time. This CMV virus can be transmitted by many aphid species with very high transmission efficacy that also plays a role in spreading the virus [12]. On the other hand, low percentage of virus symptoms in the treatment of silver plastic mulch is due this mulch has the ability to reflect about 33 percent of the sunlight that hits its surface [9], which will put away the insects from the plants toward the reflected light, so escaping plants to be infected by virus carried by the aphids. Plant Height The result shows that chili plant height planted on silver plastic mulch was the highest (101.05 cm)

compared to one that planted in a black plastic mulch (83.16 cm) and controls (49.97 cm) (Table 4). Statistical analysis showed that plant height in the treatment of silver plastic mulch was higher and significantly different to black plastic mulch treatment and control. Plant height seems to be associated with symptoms that appear in an infected plant virus. Chili plants in the control treatment had the lowest average plant height compared to black and silver plastic mulch (Table 4), but the percentage of chili plants attacked by viral disease at control, was the highest among treatments (Table 3). The metabolism of plants that were showing symptoms of viral infection was impaired. The decreased production of growth hormone produced by plants, accompanied by a decrease in the amount of chlorophyll is a common effect that occurs in plants infected by the virus. This will result in disturbance of plant growth, therefore plant height [13].

TABLE 4. PLANT HEIGHT, NUMBER OF BRANCHES AND YIELD (TONS/HA) OF CHILI PLANTS PLANTED IN THREE DIFFERENT TREATMENTS (NT: NO TREATMENT OR COMTROL; MH: BLACK PLASTIC MULCH AND SILVER PLASTIC MULCH). Yield (ton/ha) NT 49.97 c 10.92 b 4.87 c MH 83.16 b 14.6 a 8.89 b MP 101.05 a 14.8 a 12.43 a Note: Number followed by the same letter in the same column showed no effect among treatments on Duncan test at 5%. Treatment

Plant height (cm)

Number of primary branch

Journal of Advances in Tropical Biodiversity and Environmental Sciences, Vol. 1 No. 1, February 2017

Number of Primary Branches The highest average number of primary branches produced was from chili planted in the silver plastic mulch (14.8), followed by black mulch treatment (14.8) and the lowest was in control treatment (10.92). Statistical analysis shows that the average number of primary branches produced was not significantly differ between chilis planted in silver and black plastic mulch, but with control treatment (Table 4). Metabolic processes in the vegetative period greatly affects the process by which plants entering the generative period [13]. Effect of plastic mulch on the growth and yield determined by the balance of the light that hits the surface of the plastic used. In general, the sunlight that hits the surface of the silver mulch, large proportion will reflected back, and only a small portion is absorbed, transmitted and reaches the ground surface, while all light that hits black plastic mulch is absorbed. The capability of the plastic mulch in reflecting, absorbing and passing the light is determined by the color and the thickness [14][15][16]. Light reflected from the surface of the plastic mulch affects the lower surface of leaves of the plant, so that the light distributed evenly for photosynthesis, while the light transmitted into the subsurface plastic mulch will affect the physical, biological and chemical of the rhizosphere covered. Sunlight that passed across the surface of mulch stuck in the ground and form a greenhouse effect in a small scale [17][18]. Increasing number of primary branches per plant affect the amount of flowers formed per plant. The more primary branches are produced, more flower also formed from the axial of the branches [19]. Yields Similar to the number of primary branches produced, the yields were significantly higher in chili plants growing in silver plastic mulch (12.43 ton/ha), compared to black plastic mulch (8.89 ton/ha) and control (4,87 ton/ha) (Tabel 4). Plastic mulch has several advantages, such as it has effectiveness in protecting the soil from exposure of direct raindrops, preventing splash of soil onto the plant, preventing the nutrient leaching, maintaining soil porosity, slowing the release of soil carbon dioxide from respiration activity of microorganisms, maintain soil temperature, prevent soil water evaporation, and maintain soil organic matter content. Mulch also has important function to control the growth of weeds, which is the main competitor in of plant to gain nutrient from soil. Weeds has been found as the host of the virus, as well as has the ability to suppress viral vector insect populations [20], therefore viral infection of the plant. As a result, chili plants were well growing, indicated by increasing plant heights, number of primary branches and yeilds. Low yileds from control plants related to high number of plants were infected by viruses at the beginning of growing phase. Low amount of growth hormone was produced in plants infected by viruses, which are accompanied by a decrease in the amount of chlorophyll. This leads to disruptions in plant growth that directly

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affects the yileds [13]. The use of plastic mulch affects microorganisms activity (as a result of increases rhizosphere temperature), which are contributing to the growth and yield by increasing carbon dioxide concentrations in the planting zone [11] and the supply mocro organic matter [21]. Dark-colored plastic mulch is very effective in controlling weeds [9]. This happens because the seeds of weeds under black plastic mulch has no access to sunlight for photosynthesis, so the weeds will experience etiolation and grow weaker. This weak growth will be exacerbated by the relatively hot temperatures and high soil moisture. Hot and high moisture enviroment has a higher lethal effect to the weeds than dry heat. Other studies also found that the use of black-silvered plastic mulch consistently effective in suppressing the growth of weeds, where the weed is a major competitor of the plant in taking water and nutrients [20][22][23][24]. The advantages of the use of silver plastic mulch in producing the highest yields compared to black plastic mulch and control (no treatment), was related to its ability in reducing pest populations in chili plants, and indirectly able to reduce the incidence of viruses [20]. This is because the pests were served as vectors of the viruses [25], where indirectly, the use of silver plastic mulch can suppress viral disease, resulted in plants were growing better, so increase plant resistance [11]. Another advantage of the use of silver plastic mulch is that, it is able to reflect light so it does not illuminate the ground directly. The increasing amount of light that plants cought will increase the photosynthesis, resluts in higher carbohydrate produced [26]. This is in accordance with Harjadi (1993) who found that the amount carbohydrates will affect the size of the cell formed, because the carbohydrates produced from photosynthesis will be used in the process of cell division and enlargement of the fruit. IV

CONCLUSION

The use of silver mulch in chili crops was found the best in controlling insect viral vector compared to black plastic mulch and control, and increased in crop yields. ACKNOWLEDGMENT This study was funded by Udayana University with Letter of Agreement No.: 6417/UN14.2PNL.01.03.00/2016, dated 15 th June 2016. REFERENCES [1] Sulyo, Y. 1984. Penurunan Hasil Beberapa Varietas Lombok akibat Infeksi Cucumber Mosaic Virus (CMV) di Rumah Kaca. Laporan Hasil Penelitian, Balai Penelitian Hortikultura Lembang 1982/1983. [2] Nyana, D.N. 2012. Isolasi dan Identifikasi Cucumber Mosaic Virus Lemah untuk Mengendalikan Penyakit Mosaik pada Tanaman Cabai (Capsicum frutescens L.). Disertasi. Program Studi Ilmu Pertanian Program Pascasarjana Universitas Udayana. Denpasar.

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