Research And Analysis Of Trichoderma Spp. Related Mushroom Cultivation

In this paper. 49 strains of Trichoderma spp. which associate with green mold epidemic of edible fungi were researched. Those were isolated from pollution of the edible fungus and fruiting body. This article also classified and identified those strains by morphological and molecular method which was ITS sequence. The results showed that there were mainly Trichoderma harzianum and Trichoderma longibrachiatum in Fujian and Zhejiang provinces and so on in Chinese Mushroom farm. There were also a few Trichoderma atroviride and Trichoderma asperellum. Trichoderma species related to locations and types of edible fungi collected to certain extent, such as there were mainly T.harzianum in Zhejiang Qingyuan Lentinus edodes. There were mainly T.asperellum in Guangzhou Pleurotus Eryngium. But isolation of Lentinus edodes polluted from Fujian Pucheng were mainly T.atroviride. Molecular characteristic of 29 T.harzianum strains, 15 Trichoderma longibrachiatum strains, 3 T.atroviride strains and 2 T.asperellum strains were set out in this article. ITS were 575bp-578bp, 579bp- 583bp, 565bp-567bp and 560bp-561bp, respectively. GC content were 55.6%-56.7% ,57.2%-58%, 55.6%-56.1% and 55.8%-55.9%, respectively. which displayed the ITS length diversity of T.spp. ITS1 sequences were respectively 221bp-233bp, 248bp-255 bp, 214bp-216bp and 213bp-214bp, GC content were 55.4%-57.4 %, 57.5%-58.7% ,55.4% -55.6% and 55.6%-55.9%. ITS2 sequence were 185bp-188bp, 168bp-183bp, 189bp-192bp and 188bp, respectively. GC content were 63.1% -65.6%, 66.1%-67.9%, 62.9%-64.7% and 63.8%, respectively. All of T.spp.'s 5.8 S were 159 bps, GC content were 46.5%. ITS sequences were suitable for classification and identification marker. ITS1 sequence were not suitable. ITS2 can not be a good marker if there were homologous T.spp.. Different strains were different from each other in RAPD and biological characteristic, which showed that there were broadly diversity in T. spp. Sensitivity of Trichoderma to five kinds of fungicide which were usually used in Edible Fungi cultivation were set out in this article. T. spp. can not grow in the thiophanate-methyl and carbendazim PDA culture medium in any concentration. Chlorothalonil and Mancozeb can also restrain Trichoderma growth to some extent. But metalaxyl is not suitable for preventing Trichoderma. Results showed that different T.spp. strain had different aggressive ability and the aggression of Trichoderma toward mushroom was different from it toward Pathogens of plants. That is, the most aggressive strains of T. spp. toward mushroom maybe feebleness toward pathogens of plant, which demonstrated that the specialty of aggressive ability of Trichoderma spp. In this paper the aggressive mechanism of T. spp was also discussed, and the result showed that Trichoderma can via lying cell walls, Winding means etc. to kill edible fungi mycelia. Diffusible inhibitory substances from T.spp. inhibited mycelium growth of edible fungi and pathogen fungi of plant. These T. spp. produced chitinase, but there was no consistent association between levels of chitinase and aggressiveness. T.spp's chitinase activity were not directly related to their aggressiveness. Aggressive strongly Trichoderma strains which activity of chitinase were not necessarily high, vice versa. There were not consistent association between Chitinase activity and chitinase gene sequence and the amino acid sequence. such as T.atroviride strains 66 and 69 chitinase activity were respectively 0.0204 U / mL.min and 0.0044 U / mL.min. Different species of T. spp. can have similar chitinase sequence. Chitinase activity had nothing to do with the infestation. such as T. atroviride 52 were aggressive strongly both in plant pathogens and the edible fungus. But its chitinase activity was one of the weakest. T.spp chitinase gene and the amino acid sequence had something to do with the infection. Aggressive strong T. atroviride strain 52 and T. asperellum strain 29 were in the same clustering, so and weak aggressive T. asperellum 30 and T. longibrachiatum 33. But the common aggressive T.harzianum 45 and strong aggressive T. atroviride 66 were clustering alone. Trichoderma cellulase were not directly related to infection. Infection strong in edible fungi were Trichoderma strains 4,29,52 and 66, the common were Trichoderma strains 19,38,63 and 69, the weak were Trichoderma strains 28,30,37 and 70. But which produced cellulase in high level were Trichoderma strains 29 and 63. The middle are 19,28,30,37,38,52,66,69 and 70, the weak were T.spp. strains 4. Cellulase activity and cellulase gene sequence and amino acid sequences were unrelated to Trichoderma species. Four of the five cloned Trichoderma cellulase gene sequence were T.harzianum and one of them was T. longibrachiatum. Their gene, cDNA and amino acid sequence were high homology, which can be reached 97.3% to 100%. Cellulase gene sequence and amino acid sequence had nothing to do with the infection. The weak infection of T. harzianum strains 20 and the common strain 90 were homologous recently and clustered together. Ability of the same infection T.spp strains 44,47,65 and 90 did not come together.