Objective

2

Isolate and identify

Mycena

expand the resources of geminating fungus of

Gastrodia elata

and optimize the culture conditions of

Mycena

，in order to provide information and guidance for the production of geminating fungus of

G. elata.

Method

2

Juvenile tuber tissue mass transfer separation and purification technology was used for the separation and purification of strains，traditional morphology microscopy was used to isolate the colony mycelia spores morphological characteristics

such as identification，polymerase chain reaction（PCR） amplification rDNA （Ribosomal DNA） internal transcribed spacer（ITS） was used for sequencing analysis and further homology with NCBI database retrieval，MEGA6 software was used to establish Phylogenetic tree by the Maximum likelihood method （MaximumLikelihood，M-L）

so as to classify and identify isolated strains. At the same time，orthogonal test was used to optimize the optimal growth conditions of Mycena.

Result

2

A total of 86 strains were isolated

which belong to 21 species in 12 genera. WMMFJ，SHXG，WMM-21 and MFJ8103

were identified as

M. purpureofusca

and ZT01-6 and ZT01-8 were identified as

M. cf. purpureofusca

. The growth rate of

Mycena

in wheat bran medium was significantly higher than in PDA medium. The optimal medium composition for the growth of germinating bacteria was 100 g potato，150 g wheat bran and 20 g corn flour，100 g glucose. And 1，3，5-Trihydroxybenzene significantly promoted the growth of WMMFJ，and played a role in promoting the growth of WMM-21 and ZT01-6，and 2-Methoxyphenol promoted the growth of WMMFJ.

Conclusion

2

Six strains of

Mycena

were isolated and identified，four of them are

M. purpureofusc

a，and two of them are

M. cf. Purpureofusca

. The separation method improved the separation effect of germinating bacteria.