利用拟南芥鉴定双孢蘑菇Hsp20和Adcs基因的耐温功能

Identification Thermotolerance Function of Hsp20 and Adcs Genes fromAgaricus bisporus by Arabidopsis thaliana

双孢蘑菇2个差异表达基因热休克蛋白基因(Hsp20)和4-氨基-4-脱氧分支酸合成酶基因(Adcs)通过根癌农杆菌(Agrobacterium)介导的花序浸渍法转化拟南芥(Arabidopsis thaliana),经草铵磷筛选、聚合酶链式反应(PCR)鉴定和蛋白质印迹(Western blot)鉴定表明,Hsp20基因和Adcs基因已整合到拟南芥基因组中并过表达.对转基因植株和野生型对照进行高温胁迫,比较其耐热性差异.结果显示:经过45℃热激1h处理,Hsp20基因转化拟南芥下胚轴恢复生长,部分幼苗存活;Adcs基因转化拟南芥与野生型一样下胚轴未恢复生长,幼苗基本未存活.实验表明,Hsp20基因对蘑菇的耐热性有直接作用,而Adcs基因对蘑菇的耐热性可能无直接作用.

As two differentiated expressing genes, heat shock protein 20 （Hsp20） and 4-amino-4-deoxychorismate synthase （Adcs） were transformed into wild type Arabidopsis plants by Agrobacterium tumefaciens-mediated floral dip method, after Basra screening,PCR and Western blot detection indicated Hsp20 and Adcs gene had been integrated into the genome of Arabidopsis and overexpressed.The performance of the transformed plants and the wild type under heat stress was analyzed.The results showed that, after 1 hour stress of 45 ＊C ,the hypocotyl of Hsp20 transgenic plants can recover extending,whatls more^some seedlings of trans- genic plants can survive;however,like wild type,the hypocotyl of Adcs transgenic plants canr t recover extending and their seedlings cant t survive.Consequently,the experimental results displayed that Hsp20 gene contributes thermotolerance to Agaricus bisporus di- rectly whereas Adcs gene probably not.