Study On The Diversity Of The Endophytic Fungi And The Rhizosphere Fungi Of Drepanostachyum Luodianense(Yi Et R.S Wang) Keng F.

In this paper, taking Karst area of endemic and endangered species of Drepanostachyum luodianense(Yi et R. S. Wang) Keng f. as the research materials, based on establishment of the method of optimized separation of the Endophytic Fungi of D. luodianense, we isolated 103 strains of endophytic fungi, by tissue isolation and tissue fluid grinding extraction from bamboo of root, stem, and leaf and through the classical morphological and molecular biological methods to identify them. In addition we further explore the correlation of endophytic fungi in the roots and rhizosphere soil fungi. The main results are discussed as follows:1.Establishment of the systematic method of isolation of endophytic fungi from D. luodianense. In this experiment, we take sodium hyp℃hlorite solution as main disinfectant in disinfection of d.luodianense tissue surface, and set up for the sodium hyp ℃ hlorite solution concentration and sterilization time in different tissues to explore. The best optimum surface sterilization pr℃edure for l D. luodianense was 2.5%NaCIO 1min stem 2min, 2.5%NaCIO solution; root, 5% NaCIO 3min solution. In addition, through the screening of the separation medium and culture conditions, to determine the concentration of 25% PDA of culture medium, culture temperature 26 ℃ as the best conditions to isolate of Endophytic Fungi from D. luodianense. In this way, we can separate endophytic fungi from D. luodianense maximum extently.2.The system of organization of D. luodianense in endophytic fungi were isolated and identified. In this experiment, by using tissue isolation from the total 180 tested tissues 81 strains of endophytic fung were isolated and in the method of tissue fluid, 22 strains of endophytic fungi grinding extraction total from 120 isolated medium. Statistics found that, D. luodianense, the colonization of Endophytic Fungi of total endophytic fungi rate was 47.2%, the root, stem and leaf of colonization rates were 73.3%, 35% and 33.3%; the total isolation rate was 45%, the root, stem and leaf of the separation rate were 91.7%, 20% and 23.3%.The result were all the root highest, stem and leaf second, with the mature degree of plant tissue showed increasing trend. After morphological observation and 5.8S rDNA-ITS sequence analysis, 103 firngal strains were identified, which belonged to 1 subdivisions, 5 classes, 10 orders, 17 families, 19 genera and 31 kinds(including two possible new species), the genus classification level to Arthrinium Mortierella(Arthrinium), Fusarium(Fusarium), Trichoderma(Trichoderma), genus Xylaria(Xylaria) was the dominant species, accounting for 12.62% of the total isolates, 9.71%, 6.80%, respectively. The results show that the endophytic fungi of Drepanostachyum has a rich diversity.3.Diversity of endophytic fungi and distribution characteristics in D. luodianense. In a small tent, endophytic fungi and the number of species and distribution, there are obvious differences in the different tissues and showed some tissue specificity: root fungi isolation frequency was the highest, reached 61.17%, the highest frequency ammog them was Fusarium and Myrothecium; among stems the dominant bacteria of the genus were mitosporic Valsaceae and Eutypella; leaf section Arthrinium was predominant genus.The diversity index of endophytic fungi: Shannon-Wiene and Simpson index of D. luodianense were 2.800, 0.932.Different organizations in the same trend, the highest for root, stem secondly and leaves were the lowest;evenness index E is 0.604, the highest in the stem, root secondly, leaves the lowest. In addition, in different tissues of Endophytic Fungi of the similarity comparison found that the lowest similarity of roots, stems and leaves of the highest similarity.4.Through the comparison of endophytic fungi in the roots and rhizosphere soil fungi of D. luodianense, We found that the fungi are common such as Cordana, Arthrinium,Neurospora,Trichoderma and Myrothecium, which shows two groups have close relationship because of a mutual exchange and communication.Another study found that in root tissue isolated endophytic fungus strong parasitic bacteria such as Fusarium and Myrothecium are dominant but in rhizosphere soil fungi Penicillium, Trichoderma, Aspergillus, a strong class of saprophytic flora dominated,are dominant, which indicating there is also a significant difference between the two groups and suggesting that endophyte may have other sources.5.Diversity of endophytic fungi in different organs and different growth periods of D. luodianense were studied systematically in this study,which provides a theoretical basis furtherly the study and use of its resources endophytic fungi, Drepanostachyum introduction and cultivation and protection and utilization. These results indicate that fungi in root endophyte and rhizosphere soil from D. luodianense have abundant microbial diversity.