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Matsutake hyphae morphological and genetic characteristics of

Author: LiaoDeCong
Tutor: ZhangXiaoPing;LiDengZuo;ChenQiang
School: Sichuan Agricultural University
Course: Microbiology
Keywords: Tricholoma matsutake Scanning Electron Microscope Transmission Electron Microscope Genetic Characters AFLP internal trasicribed spacer Phylogeny
CLC: S646.15
Type: Master's thesis
Year: 2004
Downloads: 335
Quote: 1
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Abstract


The achievement of studies on Tricholoma matsutake was briefly reviewed in this paper, with emphasis on investigations in Tricholoma matsutake resources, identification and taxonomy, ecology, physiology, molecular biology, and biotechnology. Based on the current situation, suggestions were made to improve the research and utilization of Tricholoma matsutake in China, especially in Sichuan.Due to the great diverse topography and climate, Sichuan is extraordinarily rich in fungi e.g. Tricholoma matsutake. The western mountains of Sichuan are the center of Tricholoma matsutake in the world. It is necessary to protect the natural resources of Tricholoma matsutake in these regions. A total of 10 Tricholoma matsutake strains isolated from Yajiang, Sichuan, were analyzed by using SEM, TEM, AFLP and ITS analysis techniques. The following results were obtained.1. The ultrastructure of Tricholoma matsutake The results of scanning electron micrographs showed that the external colony surfaces of Tricholoma matsutake were very similar and composed of dense and innumerable villi, which the diameter was 1.2~3.2un;i. The ultrastructure of Tricholoma matsutake Fll was observed by transmission electron microscope, the results suggested that most hyphae were hollow, only the cell wall and some endomembrane system could be seen. In few hyphae, there was a single nuclear with clear nucleolus which located in the middle of the cell. Some cells were during the mitosis stage. Most hyphae had obvious septa, and a large amount of mitochondria.2. AFLP analysis The analysis results showed that the genetic characteristics of Tricholoma matsutake strains can be properly revealed by AFLP, and, among the 10 Tricholoma matsutake strains, the genetic aspects were very similar. At the level of 90% similarity, all strains formed a single cluster, and at the similarity of 92%, 3 groups formed. Simultaneously, the effects of quantity of the primer pairs used in the AFLP PCR were detected, and the appropriate quantity of Eco+1/Mse+1, Eco+2/Mse+2 and Eco+3/Mse+3 primer pairs used in the AFLP PCR was 1pmoL/5pmoL, 0.6pmoL/10pmoL and 0.1pmoL/8pmoL, respectively.3. 18S rDNA PCR-RFLP and ITS RFLP Furthermore, using NSland NS6, NSlandNS8, ITS5 and ITS4 primer pairs, 18S rDNA and ITS fragments of 10 Tricholoma matsutake strains isolated from Yajiang was amplified, and digested by AluI, HaeIII, HinfI and Mspl, respectively, the results showed that these strains had the same restriction patterns, which suggested that the homology among these strains were very high. The ITS sequence of the representative strain E7 was the same as these of Tricholoma matsutake strains in the genebank, which disclosed that the strains isolated from Yajiang belong to Tricholoma matsutake.

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