International Journal of Recent Academic Research ISSN: 2582-158X Vol. 02, Issue 12, pp.784-787, December, 2020 Available online at http://www.journalijrar.com

RESEACH ARTICLE

EXTRACT TEST OF WHITE ONION AND GALANGAL PRESSURE PATHOGENIC COLLECTION COLLECTOTRICHUM SP. CAUSES OF ANTHRACNOSA DISEASE IN PLANTS 1,*Dyah Roeswitawati, 2Ulfa Waldani and 3Dian Indratmi

1,3 2 Muhammadiyah University of Malang ARTICLE INFO Student atABSTRACT Agrotechnology Departement of Muhammadiyah University of Malang

Article History: Anthracnose disease caused by the pathogenic fungus Colletotrichum sp. in plants has a wide distribution. The Received 16th September, 2020 pathogen attacks several plants, especially horticultural crops, for example, chili and onions. So far, many farmers Received in revised form rely on chemical pesticides that are considered easy and practical to use, but negatively affect humans, pets, and 29th October, 2020 Accepted 17th November, 2020 the environment. For this reason, there are currently a lot of studies on plant-based pesticides, including garlic Published online 30th December, 2020 and galangal extracts that have the potential to act as agents for controlling microbial pathogens. The ability of both Plant extracts is able to inhibit the growth of pathogenic microbes widely, including viruses, bacteria, protozoa, and fungi. The purpose of this study was to examine the ability of garlic extract and galangal to inhibit the growth of the pathogen Colletotrichum sp. causes of anthracnose disease in plants in vitro. The research was conducted at the Agronomy Laboratory of the Faculty of Agriculture and Animal Husbandry, University of Muhammadiyah Malang, from September to December 2018. The research was arranged using a simple Completely Randomized Design and was repeated 4 times, then tested using a 5% Duncan level test.The results showed that the treatment of garlic extract and galangal were able to inhibit the growth and development of the *Corresponding author: pathogenic fungus Colletotrichum sp., Respectively by 38.14%, and 10.18%. Dyah Roeswitawati Key Words: Colletotrichum sp., Extract, Galangal, Garlic. Copyright © 2020, Dyah Roeswitawati et al. This is an open access article distributed under the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

INTRODUCTION Fungus Colletotrichum sp. is pathogenic and causes anthracnose disease in several types of plants, such as chili Chemical pesticides are widely used because they are cheaper plants and onion plants (Ensiklopedia, 2017) . The fungus and have a faster effect, but they have a negative impact on the produces conidia in large quantities, which are formed on the environment, causing resistance to chemical pesticides, surface of spots on infected plant parts, and the conidia are pesticide residues in agricultural products, workers poisoning easily released when in the wind or when exposed to rain (Murray, 2020; Kardinan, 2005). To overcome this, currently, splashes(Bessey, 1979; Semangun 2007) . Signs of leeks that a lot is learned about plant pesticides. Vegetable pesticides are are attacked by anthracnose disease are the lower leaves fall, considered environmentally friendly because they are easily the base of the leaves shrink, dark in color, and plants die biodegradable, safe for humans and pets. Vegetable pesticides suddenly. The purpose of this study was to examine the ability also play a role in government efforts to develop organic of garlic extract and galangal extract to inhibit the growth of agriculture, where the use of synthetic chemical pesticides is the pathogen Colletotrichum sp. causes of anthracnose disease prohibited(Badan, 2012, Khedr, 2018)]. The results showed in plants in vitro. that the use of garlic extract was able to inhibit the growth of pathogenic fungi and pests(Hermawan, 2003, Hasnah, 2010). Likewise, galangal (Alpinia galanga) rhizome water extracts MATERIALS AND METHODS on the growth of the fungus Aspergillus spp. Aflatoxin and Fusarium moniliforme producers are also able to suppress the The research was conducted at the Agronomy Laboratory of growth of Colletotrichum sp. in soybean seeds(Direktorat, the Faculty of Agriculture and Animal Husbandry, University 2018, Handajani, 2008). It was also stated that extracts from of Muhammadiyah Malang, from September to December garlic and galangal have potential as microbial agents 2018. The experiment used a completely randomized design (Anandita, 2011; national 2014).Its ability to inhibit the growth and 4 replications, the treatment was as follows: P1, garlic of microbes is very broad, including viruses, bacteria, extract concentration of 5%; P2, galangal extract concentration protozoa, and fungi (Solehah, 2012). of 5%; P3, a combination of galangal extract and garlic 5% ratio of 1: 1; P4, a combination of galangal extract and garlic 5% ratio of 1: 2; P5, a combination of galangal extract and *Corresponding author: Dyah Roeswitawati, garlic 5% ratio of 2: 1; P6, synthetic fungicide; Q7, without Muhammadiyah University of Malang. treatment. 785 Dyah Roeswitawati et al. Extract test of white onion and galangal pressure pathogenic collection collectotrichum sp. causes of anthracnosa disease in plants

Research Implementation: Isolation of the fungus are presented in Figure 1. In figure 1, the morphology of the Colletotrichum sp.done by taking from the leaves of leek pathogenic fungal colonies of Colletotrichum sp. the PDA plants that are attacked by anthracnose disease, amounting to 1 media is white (figure 1C), and if observed under a microscope cm. Sterilization is done by soaking the leaves in Clorox 2% with a magnification of 400x (figure 1B), the conidial shape of for 2 minutes, then with 70% alcohol and rinsed with sterile the pathogenic fungus is oval-shaped like a capsule with aquades (Ramadhan, 2017) .Then planted on PDA (potato patches of oil in the middle (looks like an eye). dextrose agar) media and incubated for 7 weeks. Observations were made by looking at the structure of mycelium, spores or Growth Diameter Colletotrichum sp: The results of the conidia (Sanjaya et al, 2010).Observations were made to analysis of variance showed that the treatment of garlic extract, identify the fungus Colletrotichum sp that causes anthracnose galangal or a combination of both had a very significant effect disease of the leek plants. Garlic and galangal extraction, on the growth of Colletotrichum sp. at the age of 4 DAI to 10 extraction method uses a maceration (Istiqomah 2013), that is DAI, but no significant effect on the growth of Colletotrichum 1 kg of crushed garlic added 1 l of distilled water. Extraction is sp. at the age of 11 DAI.To find out the difference between done by cold (maceration), maceration process is carried out each treatment Duncan test was carried out at a 5% level and for 24 hours. The same thing is done for galangal too. The the results are presented in Table 1. Table 1 shows the average results of the immersion process are then filtered to separate colony diameter of Colletotrichum sp. due to the treatment of the extract from the impurities and continue with the garlic extract showed no significant difference compared to evaporation process to evaporate the solvent used. synthetic fungicide treatment, whereas compared to galangal extract treatment showed significant differences in Inhibition Test, done by the method of making solvent media, observations of 4 DAI to 10 DAI. The combination treatment which is the extract of garlic and galangal dissolved into the of garlic and galangal extract with various comparisons did not media Potato Dextrose Agar (PDA)(Suaib, dkk, 2016).The show any significant difference in the diameter of the colonies diluted PDA media was added with garlic extract and galangal of Colletotrichum sp. at each age of observation with a concentration of 5% (95 ml PDA media + 5 ml extract). After that, the media was put into 20 ml Petri dishes. Isolate Percentage of Obstacles of Colletotrichum sp: Results of the Collectotrichum sp. purification results perforated with a analysis of variance showed that the treatment of garlic extract, diameter of 1 cm. Next is the fungus inoculation galangal or a combination of both had a very significant effect Collectotrichum sp. in the media that has been mixed with the on the percentage of inhibition of the pathogenic fungus extract. The position of the inoculum should be in the middle Colletotrichum sp. at the age of 4 DAI to 10 DAI, but each of the petri dish which is 4.5 cm from the right, left, top, and treatment had no significant effect on the percentage of bottom. Petri dishes used are Petri dishes with a diameter of 9 inhibition of pathogenic fungus Colletotrichum sp. at the age cm. of 11 DAI. To find out the difference between each treatment Duncan test was carried out at a 5% level and the results are The variables observed in this study were: The diameter of presented in Table 2. Table 2 shows the average colony Colletotrichum sp.Observation and measurement of diameter diameter of Colletotrichum sp. due to the treatment of garlic are carried out or until the control meets the petri dish. extract showed no significant difference compared to synthetic Diameter measurement using millimeter block paper or a ruler. fungicide treatment, whereas compared to galangal extract The method of measurement on a Petri dish is based on the treatment showed significant differences in observations of 4 following formula: HSI to 10 HSI. The combination treatment of garlic and galangal extract with various comparisons did not show any = , note:D = diameter of Collectotrichum sp. significant difference in the diameter of the colonies of Colletotrichum sp. at each age of observation. d1 = vertical diameter of Collectotrichum sp. d2 = horizontal diameter of Collectotrichum sp. DISCUSSION Percentage of the relative resistance of Colletotrichum sp.The ability of the relative inhibition of treatment to the growth of Based on the results of the insulation Collectotrichum sp. from the fungus Colletotrichum sp. was calculated until the fungus leek plants showing symptoms of anthracnose disease, a had grown. The percentage of obstacles is calculated according conidia morphology was obtained in the form of a capsule to the formula[Joshi et al. 2013 in [21] are as follows: whose middle part looked like an oil beam in the middle of the conidia (figure 1B). Mushroom Collectotrichum sp. generally has a cylindrical conidium with blunt edges, sometimes a HR = x100%, note:HR = relative obstacles rather long-shaped with a slightly rounded tip with a rather dk = control diameter kontrol narrow base, not insulated, nucleated (Ensiklopedia, 2017) . In dp = treatment diameter accordance with the results of the study (Spolehah, 2012; Bessey, 1979), where the characteristics of conidia The influence of a fungicide is assessed from the categories Collectotrichum sp. identified as having patches in the middle proposed by [22] as follows: 0 = no effect,> 0-20% = very less with capsule shape. The morphology of the colony influential,> 20-40% = less influential,> 40 - 60% = macroscopically shows a collection of mycelium which is sufficiently influential ,> 60 - 80% = influential,> 80% = very slightly pinkish white (figure 1C). This was stated by (Desta, influential. 2017; Nurul 2017) that the pathogenic fungus colony Collectotrichum sp. white, and when there is an ongoing RESULTS infection the colony has a pink color, which then turns dark brown, and finally black. The mean percentage of inhibition Based on the identification of pathogenic fungi Colletotrichum showed that garlic extract treatment had the highest average sp. isolated from leek plants showing symptoms of anthracnose percentage among the other treatments that was 38.14%, 786 International Journal of Recent Academic Research, Vol. 02, Issue 12, pp.784-787, December, 2020

Figure 1. Morphology of fungal pathogens Colletotrichum sp .: (A) hypha Collectotrichum sp. 400x magnification; (B) conidia Collectotrichum sp. 400x magnification; (C) pathogenic colonies Collectotrichum sp. on PDA media

Table 1. The average diameter of pathogenic fungal colonies of Colletotrichum sp. due to the treatment of galangal extract and garlic at each age of observation (days after inoculation / DAI)

Treatment Diameter of pathogenic fungal colonies of Colletotrichum sp.(cm) 4 DAI 5 DAI 6 DAI 7 DAI 8 DAI 9 DAI 10 DAI 11 DAI Onion (O) 1,45 a 2,34 a 3,13 a 4,44 a 5,26 a 6,36 ab 7,15 a 8,08 a Galangal (G) 3,49 bc 4,43 c 5,21 b 5,85 bc 6,85 b 7,45 bc 7,95 abc 8,75 a G+O (1:1) 3,00 b 4,18 c 5,05 b 6,10 c 6,91 b 7,57 c 8,16 bcd 8,89 a G+O (1:2) 3,03 b 4,26 c 5,25 b 6,18 c 7,33 b 7,81 c 8,41 cd 9 a G+O (2:1) 3,18 b 4,33 c 5,15 b 6,21 c 7,15 b 7,88 c 8,49 cd 9 a Synthetic fungicide 1,90 a 2,51 b 3,68 a 4,80 ab 5,53 a 6,25 a 7,43 ab 8,38 a Control 3,73 c 4,91 c 5,68 b 6,73 c 7,59 b 8,53 c 8,95 d 9 a Note: the numbers followed by the same letter in the same column show no significant difference based on the Duncan test of 5%.  DAI : daysafterinoculation  G: Galangal, O: Onion

Table 2. The average percentage of inhibition of pathogenic fungi Colletotrichum sp. due to the treatment of garlic extract and galangal at each age of observation (days after inoculation / DAI)

Treatment Percentage of inhibition of pathogenic fungi Colletotrichum sp. (%)

4 DAI 5 DAI 6 DAI 7 DAI 8 DAI 9 DAI 10 DAI 11 DAI Onion (O) 61,14 d 52,57 c 44,40 c 33,31 c 30,93 cd 24,38cd 20,24bc 8,09 a Galangal (G) 8,88 b 9,89 b 8,10 b 12,97bc 9,85 b 10,37 b 11,20 b 0 a G+O (1:1) 18,63 c 14,83 b 11,11 b 9,26 b 8,90 bc 9,00 bc 8,84 bc 0 a G+O (1:2) 17,61 c 12,97 b 7,25 b 8,18 b 3,41 b 8,55 bc 5,98 b 0 a G+O (2:1) 13,62bc 11,64 b 8,89 b 7,50 b 5,71 bc 7,89 bc 5,16 b 0 a Synthetic fungicide 48,68 d 49,09 c 35,02 c 28,44 c 27,20 d 24,72 d 17,03 c 15,48 a Control 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0.00 a 0.0 a 0 a Note: the numbers followed by the same letter in the same column show no significant difference based on the Duncan test of 5%.  DAI : daysafterinoculation  G: Galangal, O: Onion 

Pure culture of Colletotrichum Pure culture of mushroom Pure culture of Pure culture of Colletotrichum sp. on PDA + synthetic Collectotrichum sp. on PDA + Colletotrichum sp. on PDA sp. on PDA + galangal extract fungicide media garlic extract media media without treatment media (control)

Figure 2. Test of garlic extract and galangal extract on the inhibition of pathogenic Colletotrichum sp. On PDA media: (A) Colletotrichum sp. + fungicide; (B) Colletotrichum sp. + on PDA media + garlic extract; (C) Colletotrichum sp. on PDA media without treatment / control; (D) Colletotrichum sp. on PDA + galangal extract media 787 Dyah Roeswitawati et al. Extract test of white onion and galangal pressure pathogenic collection collectotrichum sp. causes of anthracnosa disease in plants

followed by synthetic fungicide treatment that was 32.88%. n=com_content&view=article&id=78&Itemid=190. Galangal treatment also has a resistance of 10.18%.This is Diakses 02 Januari 2019. because garlic and galangal contain compounds that can inhibit Handajani, Noor Soesanti dan Tjahjadi Purwoko.2008. the growth of pathogens. Garlic contains allicin compounds Aktivitas ekstrak rimpang lengkuas(Alpinia where if garlic is destroyed the allicin compound will be galanga)terhadap pertumbuhan jamur Aspergillus spp. converted into allicin compounds (Sanjaya, et. al. 2010; Siaub penghasil aflatoksin danFusarium moniliforme.J. et al, 2016). Allicin is a major component of the enzymatic Biodiversitas9(3): 161-164. degradation results from flavor-forming precursors. Anandita,D. 2011.Ekstrak bawang putih (Allium sativum) Furthermore, it was stated that the content of Allicin in garlic menurunkan jumlah leukosit pada mencit model sepsis can inhibit the development of fungi. 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