Diversity of endophytic fungi derived from some potential medicinal plants

Abstract

Indonesia has high medicinal plant diversity, however, there was limited study on endophytic fungi associated with them, particularly on potential medicinal plant jahe merah (Zingiber officinale Roscoe), jati belanda (Guazuma ulmifolia Lamk), local pegagan (Centella asiatica L), malaysian pegagan, and temulawak (Curcuma xanthorrhiza Roxb). As traditional medicines, the society use certain medicinal plant organ such as leaf, stem, rhizome and root as functional organ in traditional remedy because it contains high bioactive compounds. Some endophytic fungi had been reported to produce bioactive compound including anti-microbe substances. Role of endophytic fungi in production of bioactive compound of medicinal plant as their host is significant, however, diversity and distribution of the fungi in the host plant are not sufficiently studied particularly for the five potential medicinal plant mention above. The aims of the research were: 1). To obtained the endophytic fungi isolates from the functional organ of jati belanda, local pegagan and malaysian pegagan type, and jahe merah, and from whole of jahe merah plant organs, 2). To study the diversity of endophytic fungi by using a combination of morphological characteristic, molecular analysis of sequence DNA generated from ITS rDNA region, and chemistry characteristic by FTIR, 3). To study the potency of endophytic fungi which have activity as biological control against F. oxysporum. The medicinal plants used were divided into two groups. The first group was medicinal plants that used only their functional organ that is leaf of jatibelanda, leaf of local pegagan and malaysian pegagan, and rhizome of temulawak. The second group was jahemerah plant. In this group all plant organs both functional and non-functional such as leaf, stem, rhizome and root were used as source of fungal isolation. In the first group, all functional organ of plants studied harbored endophytic fungi. There were 13 isolates of the endophytic fungi obtained from leaf of jati belanda and each of leaves of pegagan local and malaysian cultivars, and rhizome of jahe merah was occupied by five isolates. The fungi were identified as Aspergillus versicolor, Aspergillus sydowii, Aspergillus terreus, Colletotrichum gloeosporioides and its teleomorphic stage Glomerella cingulata, Colletotrichum higginsianum, Curvularia affinis, Diaporthe phaseolorum, Engyodontium album, Fusarium solani, Lasiodiplodia theobromae, Leptosphaerulina australis, Mycoleptodiscus indicus, Pseudocercospora cruenta, Stagonosporopsis cucurbitacearum, Talaromyces assiutensis. Colletotrichum gloeosporioides and its teleomorphic stage Glomerella cingulata were dominant endophyte fungi and were found in all plants study. Endophytic fungi derived from functional organ of jati belanda, temulawak, pegagan local and malaysian cultivars had inhibitation activity against F. oxysporum with the ranges of inhibition value 6.0-78.9%. Talaromyces trachyspermus JBd10, Glomerella cingulata JMr2, and Fusarium solani JMd14 statistically had the highest inhibitation value, while Mycoleptodiscus indicus PMd1 and Engyodontium album JBd13 had the lowest inhibitation value. Thirty isolates of endophytic fungi having different colony characteristics were obtained from Zingiber officinale, six isolates were derived from functional organ of Zingiber officinale and twenty four isolates were derived from nonfunctional organ of Zingiber officinale, i.e. leaf, rhizome, root, and stem. Each of root, stem, and leaf was occupied by eight isolates. Fungal identification by using a combination of morphological characteristic, molecular analysis of sequence DNA generated from ITS rDNA region, and phylogenetic analysis resulted Acremonium macroclavatum, Beltraniella sp., Cochliobolus geniculatus and its anamorphic stage Curvularia affinis, Fusarium solani, Glomerella cingulata and its anamorphic stage Colletotrichum gloeosporoides, Lecanicillium kalimantanense, Myrothecium verrucaria, Neonectria punicea, Periconia macrospinosa, Rhizopycnis vagum, Talaromyces assiutensis, and one mycelia sterilia JMd9. C. gloeosporoides and its teleomorphic stage G. cingulata were dominant endophyte fungi in jahe merah. R. vagum was found specifically on root whereas L. kalimantanense and M. verrucaria were found on stem of red ginger plant. A. macroclavatum was found specifically in bellow ground organ, whereas Curvularia was determined from shoot or above ground organ of red ginger plant. Based on moleculer analysis, chemistry characteristic by FTIR, and antagonistic assay, the isolates JMa5 and JMd14 were belong to the same species, namely F. solani but they were differ in strain. Similarly JMr2, JMd4, JMd5, and JMd12 were belong to the same species G. cingulata but they were differ in strain. Other while, C. geniculatus JMbt9 and JMd13, G. cingulata JMd12 and JMr2. Otherwhile, G. cingulata JMbt13 and G. cingulata JMd4 were belong to the same species and also the same strain that spreaded in leaf and stem of jahe merah. Similarly A. macroclavatum JMa6 and A. macroclavatum JMr7. The endophytic fungi lived in the different organs of host plant may be belong to the same species and the same strain but also belong to the same species but in the different strain. All jahe merah plant organs harbor diverse endophytic fungi and their inhibition effects on growth of F. oxysporum were varied with the inhibition value range from 1.4 to 68.8%. C. affinis (JMbt7) and F. solani (JMd14) had significantly highest antagonistic activity with the value above 65%; and R. vagum (JMa4) and C. geniculatus (JMbt9) had significantly lowest antagonistic activity with the value up to 10%. This finding showed that not only rhizome of red ginger inhabited by endophytic fungi with high antifungal activities, but also other organs.