羽衣甘蓝BoWRKY10基因克隆及其转基因烟草耐旱性研究  

Drought tolerance of heterologous expression of BoWRKY10 gene from kale in tobacco

作　　者：赵红启[1] 杨勇[1] 闫艳[1] 范杰[1] 李鸿雁 ZHAO Hong-qi;YANG Yong;YAN Yan;FAN Jie;LI Hong-yan(Garden Center of Huanghuai University,Zhumadian,Henan 463000,China;School of Biological and Food Processing Engineering,Huanghuai University,Zhumadian,Henan 463000,China)

机构地区：[1]黄淮学院园林中心,河南驻马店463000 [2]黄淮学院生物与食品工程学院,河南驻马店463000

出　　处：《西南农业学报》2022年第11期2478-2484,共7页Southwest China Journal of Agricultural Sciences

基　　金：河南省科技攻关项目(182102110305)。

摘　　要：【目的】克隆羽衣甘蓝(Brassica oleracea var.acephala DC)转录因子基因BoWRKY10,解析羽衣甘蓝BoWRKY10转录因子的抗旱调控机制。【方法】通过RT-PCR从羽衣甘蓝叶片cDNA中扩增编码WRKY的BoWRKY10基因并转入烟草中。采用实时荧光定量PCR技术检测该基因在干旱胁迫下的表达模式。【结果】干旱10 d后复水,转基因株系存活率显著高于野生型(WT)。种子萌发试验表明,正常1/2 MS培养基上,转基因株系OE3、OE4和OE6发芽率和野生型之间没有明显的差异。然而在15%PEG 4000培养基上OE3、OE4和OE6萌发率分别为65.9%、75.4%和84.3%,野生型仅为22.5%,且14 d后转基因株系根长显著高于野生型。同时,转基因株系ABA含量、脯氨酸含量、可溶性糖含量、POD、CAT和SOD活性较高,而MDA和H_(2)O_(2)含量比野生型显著降低,干旱胁迫响应基因NtSOD、NtAPX、NtCAT、NtGST、NtSPSAt和NtNCED1在OE3转基因株系中表达量显著高于野生型。【结论】羽衣甘蓝中的转录因子BoWRKY10可以显著提高烟草耐旱性,BoWRKY10可作为植物耐逆基因工程的优良基因源。【Objective】The present paper aimed to clone the transcription factor gene BoWRKY10 of kale(Brassica oleracea var.acephala DC)and study the drought tolerance regulation mechanism of the gene.【Method】The gene BoWRKY10 was amplified from the cDNA of kale leaves by RT-PCR and transferred into tobacco.The expression pattern of the gene under drought stress was detected by real-time fluorescence quantitative PCR.【Result】The survival rate of transgenic lines were significantly higher than that of non-GMO wild types(WT)after 10 days of drought.There was no obvious difference between transgenic lines(OE3,OE4 and OE6)and WT in terms of germination rate in 1/2 MS culture medium,however,in medium containing 15%PEG4000,germination rate of OE3,OE4 and OE6 were 65.9%,75.4%and 84.3%,respectively,while the one of WT was 22.5%.In medium containing 15%PEG4000,the root length of transgenic lines was significantly higher than that of WT after 14 days.Meanwhile,ABA content,proline content,soluble sugar content,POD,CAT and SOD activities were higher,while MDA and H_(2)O_(2)content were significantly lower than those of WT.The expression level of drought responsive gene in OE3,like NtSOD,NtAPX,NtCAT,NtGST,NtSPSA and NtNCED1,was significantly higher than that of WT.【Conclusion】Expression of BoWRKY10 from kale remarkably improved the tolerance of tobacco to drought stresses in which the pleiotropy could be beneficial gene resource of gene engineering about resistance to stress in plant.

关 键 词：WRKY 抗旱性 转基因烟草 羽衣甘蓝 

分 类 号：S1[农业科学—农业基础科学]