杨树类锌指基因ZFL的功能分析  

Functional Characterization of a Zinc-finger-like Gene from Populus trichocarpa

作　　者：赵学彩[1] 郑唐春[1] 臧丽娜[1] 曲冠证[1] 

机构地区：[1]东北林业大学林木遗传育种国家重点实验室,黑龙江哈尔滨150040

出　　处：《林业科学研究》2013年第5期562-570,共9页Forest Research

基　　金：国家自然科学基金项目(31070595)资助

摘　　要：以毛果杨叶片cDNA为模板,采用PCR技术分离出杨树ZFL基因,序列分析结果表明该基因序列开放读码框315 bp,共编码104个氨基酸,推导的蛋白质分子量为11.202 kDa,理论等电点9.83,命名为PtrZFL。对ZFL基因进行实时荧光定量PCR表达分析,结果表明:甘露醇、NaCl、H2O2、ABA、低温胁迫都能诱导PtrZFL基因的表达,且PtrZFL表达量在ABA处理3 h时达到最高,然后随处理时间延长而逐渐降低;低温(4℃)胁迫在6 h后能显著诱导PtrZFL基因表达,并随着低温处理能一直保持较高水平的表达。根据毛果杨基因组信息设计引物,获得了PtrZFL基因上游1 000 bp的启动子序列,序列分析结果表明该启动子包含有多个与胁迫相关的元件,如抗冻、缺水、抗寒、脱落酸响应元件ABRE、MYB和WRKY。GUS活性检测发现,该启动子在转基因拟南芥整株中都有表达,但在根部和成熟叶片中表达较强,其他位置表达微弱。In the present study, a zinc-finger-like cDNA （ZFL） was isolated from the cDNA of Populus trichocarpa leaves, named PtrZFL. The open reading frame （ORF） of the gene is 315 bp in length and encodes a predicated polypeptide of 104 amino acids with a molecular weight of about 11. 202 kDa, and the theoretical isoelectric point of which is 9.83. The expression of PtrZFL in P. trichocarpa in response to abiotic stress was studied by QRT-PCR, the results showed that the PtrZFL was response to treatments of mannitol, NaC1, H202, ABA and cold （4~C）. The PtrZFL expression level was the highest 3 hours after treated by ABA and then decreased. The PtrZFL was rap- idly and highly induced at the 6th hour when exposed to low temperature of 4~C and kept high expression level with the continuing treatment. A 1000 bp sequence upstream of PtrZFL gene was cloned from the genomic DNA of P. trichocarpa by PCR based on the poplar genome sequence. Sequence analysis showed that the promoter of the PtrZ- FL gene contained stress-induced c/s-acting elements （including cold, dehydration, frozen and ABA responsive ele- ments ABRE, MYB and WRKY）. The GUS activity was almost detected in whole plant of transgenic Arabidopsis, suggesting that this promoter could control the gene expression in the whole plant. Moreover, strong activity was de- tected in roots and mature leaves, and only weak expression could be detected in other organs.

关 键 词：杨树 PtrZFL QRT-PCR 启动子 胁迫 

分 类 号：Q786[生物学—分子生物学] S718.47[农业科学—林学]