天山雪莲再生体系的优化及转sikPIP2;7基因的研究

Optimization Regeneration System of Saussurea involucrate Kar.et Kir and Research on Overpression of sikPIP2;7

以天山雪莲幼苗叶片为外植体进行再生体系的优化,经农杆菌介导法转化获得转正义sikPIP2;7雪莲植株,对转基因雪莲苗进行低温处理并进行抗逆分析。结果表明,诱导不定芽最佳植物激素组合为MS+0.5mg/L ZT+1.0mg/L NAA;生根阶段活性炭最佳添加量为0.5g/L,蔗糖最佳添加量为20g/L。RTPCR分析表明,在0℃胁迫下转基因植株sikPIP2;7表达量增加;低温胁迫下转基因植株丙二醛(MDA)质量摩尔浓度、相对电导率(REC)、过氧化氢(H2O2)清除速率和超氧化物岐化酶(SOD)活性显著优于对照植株,说明sikPIP2;7过表达提高转基因雪莲抗低温胁迫的能力。

The leaves of Saussurea involucrate Kar.et Kir were used as explant,to optimize regeneration system of Saussurea involucrate,and the foreign gene sikPIP2;7 was transformed via Agrobacterium tumefaciens.The transgenic Saussurea involucrate were tested tolerance to chilling stress.The results showed that the best hormone combination medium of bud induced was MS＋0.5mg/L ZT＋1.0mg/L NAA;the suitable roofing culture medium was adding the amount of activated carbon in0.5g/L and 20g/L sucrose;the mRNA levels of sikPIP2;7in the leaves of transgenic plants increased under the chilling conditions.Compared with the wild type（WT）,transgenic Saussurea involucrate plants had lower malondialdehyde（MDA）molarity and the relative electrical conductivity（REC）,and higher capability for scavenging hydrogen peroxide（H2O2）and superoxide dismutase（SOD）.These results indicated that overpression of sikPIP2;7 enhanced chilling tolerance of transgenic Saussurea involucrate plants.