Efficient transformation and expression of gfp gene in the edible mushroom Pleurotus nebrodensis  

作　　者：Junfang Lin Mingyao Zheng Jie Wang Wei Shu Liqiong Guo 

机构地区：[1]Department of Biotechnology, College of Food Sciences, South China Agricultural University, Guangzhou 510640, China [2]Institute of Biomass Research, South China Agricultural University, Guangzhou 510640, China [3]Test and Experimental Center, South China Agricultural University, Guangzhou 510640, China

出　　处：《Progress in Natural Science:Materials International》2008年第7期819-824,共6页自然科学进展·国际材料（英文版）

基　　金：supported by National Natural Science Foundation of China(Grant No.30371000 and 30671457);the Natural Science Foundation of Guangdong Province(032239);the Science and Technology Program of Guangdong Province,China(2004B20101001).

摘　　要：An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleurotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a ＂reporter＂ gene, green fluorescent protein gene can be successfully stably exoressed in this Pleurotus species.An efficient transformation method mediated by PEG-protoplasts was developed for the newly commercial edible mushroom Pleu- rotus nebrodensis. Two plasmids were used to co-transform protoplasts of P. nebrodensis. One plasmid is pAN7-1 containing a positive selectable marker gene hph conferring hygromycin B resistance. Another plasmid is pBlue-GFP containing a reporter gene gfp conferring green fluorescent protein. PCR and Southern blot analysis showed that hph gene or/and gfp gene were integrated into the genome of P. nebrodensis transformants. The transformation efficiency of the positive selectable marker gene hph was 3 transformants per microgram of plasmid pAN7-1 DNA, which was about 30 times higher than that previously reported in thoroughly studied Pleurotus species such as Pleurotus ostreatus. The transformation efficiency of the reporter gene gfp was 9 transformants per microgram of plasmid pBlu-GFP DNA. The co-transformation efficiency was 23.68%. This is the first report that a "reporter" gene, green fluorescent protein gene can be successfully stably expressed in this Pleurotus species.

关 键 词：Pleurotus nebrodensis Protoplast-mediated transformation Green fluorescent protein GFP expression 

分 类 号：S646[农业科学—蔬菜学] Q78[农业科学—园艺学]