低温胁迫下草菇甘油‐3‐磷酸酰基转移酶基因表达变化的研究  

Expression of glycerol‐3‐phosphate acyltransferase gene in Volvariella volvacea during low temperature stress

作　　者：姜威[1,2] 赵妍[2] 汪虹[2] 冯爱萍[2] 陈明杰[1,2] 

机构地区：[1]南京农业大学生命科学学院,江苏南京210095 [2]上海市农业科学院食用菌研究所上海市农业遗传育种重点开放实验室农业部南方食用菌资源利用重点实验室国家食用菌工程技术研究中心,上海201403

出　　处：《菌物学报》2014年第2期334-340,共7页Mycosystema

基　　金：上海市科技兴农重点攻关项目[编号:沪农科攻字(2011)第1‐2号];上海市农业科学院青年科技基金[编号:农青年科技2012(12)]

摘　　要：以草菇低温敏感型V23菌株与耐低温型VH3菌株为试验材料,将二者菌丝体置于冰浴中进行不同时间的低温胁迫处理。首先提取RNA,反转录为cDNA,然后构建含有微管蛋白(tubulin,TUB)基因片段和甘油‐3‐磷酸酰基转移酶(glycerol‐3‐phosphate acyltransferase,GPAT)基因片段的质粒,最终对GPAT基因在低温胁迫不同处理时间下的表达进行定量。结果表明,耐低温型的VH3菌株,在低温处理2h时,GPAT基因相对表达量上升,4h,表达量下降,6h上升,之后逐渐下降。低温敏感型V23菌株,在低温处理2h时,表达量下降,4h,表达量上升,此后逐渐下降;除低温处理4h外,V23菌株的GPAT基因表达量始终低于VH3菌株,初步推测GPAT基因的高表达与草菇的耐低温能力相关。The mycelia of two Volvariella volvacea cold-resistant strains, V23 （cold-sensitive） and VH3 （cold-tolerant）, were exposed to cold stress （0℃） for 0, 2, 4, 6, 8 and 10h, respectively. RNA was extracted from mycelia and cDNA was synthesized by reverse transcription reaction with total RNA. Plasmid containing tubulin （TUB） and glycerol-3-phosphate acyltransferase （GPAT） gene fragment were constructed. The expression of glycerol-3-phosphate acyltransferase gene was quantified by real-time PCR （RT-PCR）. Results showed that the relative expression of GPAT in cold-tolerant strain VH3 increased in 2h, decreased in 4h, and increased again in 6h, then decreased gradually. For cold-sensitive strain V23, the relative expression of GPAT decreased in 2h, and increased in 4h, then decreased gradually. The expression of GPAT gene in V23 was lower than that in VH3 all along except that the strains were treated under cold stress for 4h. The better performance of strain VH3 under low temperature stress may be associated with higher expression of GPAT gene.

关 键 词：草菇 低温 甘油-3-磷酸酰基转移酶基因 实时荧光定量PCR 

分 类 号：S646.13[农业科学—蔬菜学]