Study on the characteristics of present cultural strains of Lentinula edodes and Pleurotus spp. in Taiwan

• Main Authors: Chu-Chien Su, 蘇秋建
• Other Authors: 陳隆鐘
• Format: Others
• Language: zh-TW
• Published: 2006
• Online Access: http://ndltd.ncl.edu.tw/handle/63656020206308648991

碩士 === 國立中興大學 === 植物病理學系所 === 94 === Cheaper 1 Study on the characteristics of present cultural strains of Lentinula edodes in Taiwan Lentinula edodes(Berk.) Pegler,the shiitake mushroom, is one of the most widely cultivated mushrooms worldwide. The study of the project is to investigate the characteristics of different isolates and cultivated strain 921,922,721,271 and white strain of the Lentinula edodes in Taiwn.The results indicate that the optimal medium was corn powder, the optimal C/N ratio was 20:1~40:1 and the optimal pH value was 3.5~5.6.Based on the temperature to mycelial growth, divided into high,middle,and low temperature group. In high temperature group include groupTA, in middle temperature group includes group TB, in low temperature group includes groupT C. Based on liquid cultural characteristics divided into GroupTA、TB、TC,and the optimal liquid culture medium was extract-peptone-dextrose.The optimal growth curve was twenty days and follow their characteristics of primordium, the appearance of browing and mycelium rounds. Based on the above characterization divided into GroupLA、GroupLB、GroupLC、GroupLD. Besides, in order to understand the differentence strains of L.edodes,using culture morphogenesis observation divided into GroupMA、GroupMB、GroupMC、GroupMD、GroupME. The solid culture group MA , and solid culture group MC is similar to colony morphology of liquid culture group LA . Analysis the width of mycelium and the distance of clamp connection this group is belong to group1. The solid culture group MD, liquid culture group LC, and the colony morphology character of solid culture MB group is similar to liquid culture group LD. Analysis the width of mycelium and the distance of clamp connection this group is belong to group2. The colony morphology of solid culture group ME and liquid culture group LD are not the same. Analysis the width of mycelium and the distance of clamp connection this group is belong to group3. In conclusion culture condition, comparing solid culture group and liquid culture group, we found groupA, groupF, groupJ are belong to liquid culture group LA and LC. Group I was dispersed to liquid culture group LA, LB and LC,but group D, groupE and group H are belong to LD. Analysis the width of mycelium and the distance of clamp connection are accord with before results. But the solid culture group are more complex. Becasue of the different environment affecting the morphogenesis of L.edode,somatic incompatility mating type analysis divided into compatilibity,slight compatilibity,intercompatilibity,incompatilibity etc. four level.Besides,in order to select the molecular mark to discriminate to forgein countries using DNA extraction to comparing with the strains of forgen and Taiwan strains.PCR products produced with primer ITS1 and ITS4 were visualized as a single band.The size of the PCR fragments was 720bp. Single 10-base primers were used to generate randomly amplified polymorphic DNA(RAPD) markers in the shiitake mushroom, Lentinula edodes. Ten primers produced polymorphisms in all 31 strains tested, producing 7-19 bands ranging from 0.18 to 2.52 kb.10 primers results from 27 primers RAPD profiles with amplification of 31 L.edodes isolates were select to analysis genetic similarity by UPGMA cluster analysis with computer software NTSYS-pc. We found that isolates of L. edodes have different genetic diversity. It indicated that 31 isolates of L.edodes could be divided into severn group under DNA similar coefficient of 0.68 mainly in Taiwan, some isolates is similar to foreigen countries. It indicated that 31 isolates of L.edodes could be divided into severn group under DNA similar coefficient of 0.91. It indicated that Taiwan cultivate L.edodes have rich genetic diversity.Internal transcribed spacer sequences analysis result Taiwan cultural L.edodes. and ITS2 region explose variability. Have already logged in for offering a researcher to consult in NCBI in ITS sequences to some isolates of L.edodes in Taiwan and worth further investigating. Cheaper 2 Study on the characteristics of present cultural Pleurotus spp. in Taiwan Pleurotus spp.is the main white rot fungi in forest.They have ability to decompose wood.There are about twelve varieties were cultivated worldwith. But in Taiwan,there are some varieties included P. ostreaus, P. abalones, P. cystidisus, P. eryngii were cultured. P6’s cap is 2~13.0cm across,convex at first,then becoming plane to broldly depressed when old. it’s surface dull, moist, smooth, glabrous, whitish to cream in colour. In the medium, it can form deerhorn hyphae and then form the fruit body.it’s spore elliiptical to subfusiform, smooth,about 5.5~7.4±10X3.0~4.7. The optimal fruiting temperature is 16~25℃. Somatic incompatility mating type analysis of that it was moer closed P. abalonus than P. eryngii. And the culture forms coremia , but do not form conidia on the medium or sawdust substrate.PCR products produced with primer ITS 1 and ITS 4 were visualized only a single band. The size of the PCR fragments was 680 bp. Single 10-base primers were used to generate randomly amplified polymorphic DNA (RAPD) markers in the P. spp. Four primers produced polymorphisms in all tested 30 strains, producing 2-8 bands ranging from 0.25 to 2.50 kb.Molecular genetic markers obtained with the RAPD assay can be used to differentiate isolates of P. spp. and have potential applications in mushroom breeding and strain improvement programs. Besides, we also analysis internal transcribed spacer of Pleurotus spp and login NCBI. The internal transcribed spacer explose variability in L. edodes and worth further investigating. We also used internal transcribed spacer- restriction fragment length polymorphism to analyze P. spp. It can distinguish P. spp. as seven groups and P.ostreatus(white) had the same DNA fragments with P6.Using ITS-RFLP analysis also could divide P9 and P5 into different group that we could not divide P9 and P5 into different group by somatic incompatility mating. To analysis genes involved in fruit body development of P6 , mRNA from three different developmental stages : vegetative mycelium, and fruit body. Twenty random PCR amplifications were performed with the cDNAs , whicg A, E, F, cDNA fragments from different developmental stage. Sequence analysis and database searches reveraled significant similarity with Ustilago maydis hyposis protein, Nurospora crassa filment H protein.