Isolation of endophytic fungi from benalu batu (Begonia Medicinalis) and their toxicity on Artemia Salina

S. Anam, Armini Syamsidi, Musyahidah Musyahidah, N. Ambianti, A. Widodo, M. Zubair
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Abstract

Abstract Background: Endophytic fungi are microorganisms found in plant tissues that are mutually beneficial to the host plant. They are known as sources of bioactive metabolites which possess various biological activities such as antimicrobial, anticancer, antiviral, antifungal, and antioxidant. Benalu batu (Begonia medicinalis) has been used empirically as a medicinal plant and has shown anticancer activity in breast cancer cells (T47D cells) and cervical cancer cells (HeLa cells). Objective: This study aimed to isolate the endophytic fungi from benalu batu and determine the toxicity activity and Lethal Concentration 50% (LC50) value of isolated endophytic fungi using shrimp larvae (Artemia salina). Method: Endophytic fungi were isolated from fresh parts; leaves, stems, roots of Benalu Batu. The selected parts were sterilized with ethanol 70% and sodium hypochlorite 1% subsequently placed into Potato Dextrose Agar Chloramphenicol (PDAC) then incubated at 28°C for 7 days. Isolated endophytic fungi were extracted by sonication method using ethyl acetate. The toxicity activity of isolated endophytic fungi was determined using the Brine Shrimp Lethality Test (BSLT). Results: Isolation of the endophytic fungi from benalu batu obtained 10 isolates, which the leaves part obtained 6 isolates, the stem obtained 3 isolates and the root obtained 1 isolate. The results of screening for toxicity test of 10 isolates at concentrations of 500 µg/mL shown that isolate D3, D5, and D6 gave 100% mortality. The LC50 values of isolate D3, D5, and D6 were obtained respectively in 448.03, 8.784, and 9.13 µg/mL. Conclusion: These results indicated that isolate D5 exhibits the greatest toxicity and needs to be investigated the activity for testing on cancer cells. Keywords: endophytic fungi, Begonia medicinalis, toxicity, Artemia salina
海棠内生真菌的分离及其对盐碱蒿的毒性研究
摘要背景:内生真菌是在植物组织中发现的对寄主植物互利的微生物。它们被认为是生物活性代谢物的来源,具有各种生物活性,如抗菌、抗癌、抗病毒、抗真菌和抗氧化。海棠(Benalu batu, Begonia medicinalis)作为一种药用植物,在乳腺癌细胞(T47D细胞)和宫颈癌细胞(HeLa细胞)中显示出抗癌活性。目的:本研究旨在分离batu benalu内生真菌,并测定其对虾幼虫(Artemia salina)的毒活性和致死浓度50% (LC50)值。方法:从新鲜部位分离内生真菌;Benalu Batu的叶子,茎,根。选择的部分用70%乙醇和1%次氯酸钠灭菌后,放入马铃薯葡萄糖琼脂氯霉素(PDAC)中,28℃孵育7 d。采用乙酸乙酯超声法对内生真菌进行分离提取。采用盐水对虾致死试验(BSLT)测定了分离的内生真菌的毒活性。结果:从巴豆内生真菌中分离得到10株,其中叶部分分离得到6株,茎部分分离得到3株,根部分分离得到1株。对10株分离株在500µg/mL浓度下的毒性试验筛选结果表明,分离株D3、D5和D6的死亡率均为100%。分离物D3、D5和D6的LC50值分别为448.03、8.784和9.13µg/mL。结论:实验结果表明,分离物D5具有最大的毒性,需要进一步研究其对肿瘤细胞的活性。关键词:内生真菌;海棠;毒性
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