Effective Transformation of Cordyceps militaris by Particle Bombardment

doi:10.16420/j.issn.0513-353x.2015-0145

ACTA HORTICULTURAE SINICA ›› 2015, Vol. 42 ›› Issue (9): 1837-1842.doi: 10.16420/j.issn.0513-353x.2015-0145

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Effective Transformation of Cordyceps militaris by Particle Bombardment

MAO Wen-jun*，BAO Da-peng*，ZHOU Chen-li，LI Yan，TAN Qi，and WANG Ying**

Institute of Edible Fungi，Shanghai Academy of Agricultural Sciences，Key Laboratory of Edible Fungi Resources and Utilization（South），Ministry of Agriculture，P. R.，China，National Engineering Research Center of Edible Fungi，Key Laboratory of Agricultural Genetics and Breeding of Shanghai，Shanghai 201403，China

Online:2015-09-25 Published:2015-09-25

Abstract

Abstract: An effective and convenient transformation method of Cordyceps militaris were established by means of particle bombardment. Coating microcarriers with plasmid pDHt-gpdA-GFP-bar，we transferred C. militaris CM01 by particle bombardment. After being selected by glufosinate-ammonium，two transformants were obtained under three conditions differing in the two parameters of target distance （6 cm or 9 cm） and helium pressure（7.58 × 106 Pa or 8.96 × 106 Pa）. The transformation frequency was 0.4 cfu? μg-1. PCR and southern blot results indicated that the selective marker gene Bar was integrated into the genomic DNA，a single copy or multiple copies integration，and genetically stable. Green fluorescence of the C. militaris transformant mycelia could be observed under fluorescent microscope，which indicated that the expression of green fluorescent protein was successful in the C. militaris transformants. Transformation by particle bombardment enriched the transformation methods of C. militaris. However，sample preparation and parameter setting need to be optimized，in order to improve the efficiency of transformation.

Key words: Cordyceps militaris, particle bombardment, genetic transformation, green fluorescent protein

CLC Number:

S 567
S646

MAO Wen-jun*，BAO Da-peng*，ZHOU Chen-li，LI Yan，TAN Qi，and WANG Ying**. Effective Transformation of Cordyceps militaris by Particle Bombardment[J]. ACTA HORTICULTURAE SINICA, 2015, 42(9): 1837-1842.

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Effective Transformation of Cordyceps militaris by Particle Bombardment

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