摘要
WRKY转录调控因子广泛参与植物的生长发育、生物胁迫和非生物胁迫等生理过程。本研究利用reverse transcription-polymerase chain reaction(RT-PCR)及rapid amplification of cDNA ends(RACE)技术克隆了核桃(Juglans regia L.)WRKY基因cDNA全长序列,命名为WRKY4(GenBank登录号为KC795551.1)。结果表明,克隆到的片段长度为2 174 bp,完整开放阅读框为1 554 bp,编码517个氨基酸。序列分析表明WRKY4基因含有2个WRKY保守结构域和2个C2H2锌指结构域,属于WRKY基因家族第Ⅰ类成员。利用实时荧光定量PCR分析4℃低温和自然低温条件对该基因表达的影响以及组织表达模式。结果表明WRKY4受低温诱导表达,能够在低温胁迫的早期响应这一生理过程。WRKY4在树皮(韧皮部)、雌花、花芽和叶片中均有表达,且在树皮中表达量最高,具有组织特异性。研究结果可为核桃抗寒基因工程育种提供基因资源。
WRKY transcription factors are widely involved in the physiological process of plant growth and development,biotic and abiotic stresses. By the technique of reverse transcription-polymerase chain reaction（ RT-PCR） and rapid amplification of cDNA ends（ RACE）,a full-length cDNA,designated as WRKY4（ GenBank accession number KC795551. 1） was cloned from J. regia. The 2 174 bp-length sequence of WRKY4 contained 1 554 bp open reading frame（ ORF） encoding 517 amino acids. Sequence analysis indicated that WRKY4 contained two conserved WRKY domains and two zinc finger structures（ C2H2）,belonging to GroupⅠof WRKY family. The expression of WRKY4treated with a low temperature of 4℃ and under natural conditions was analyzed by the real-time PCR technology,and the tissue expression pattern was analyzed as well. Expression analysis showed that the expression of WRKY4 was induced by cold stress,being capable of responding to the physiological process at early stage of cold stress. WRKY4gene expression was detected in bark（ phloem）,pistillate flower,flower bud and leaf,and the highest level was in bark（ phloem）,which indicated that WRKY4 gene expression had tissue specificity. These results provided gene resource for cold-resistant genetic engineering breeding of J. regia.
出处
《核农学报》
CAS
CSCD
北大核心
2014年第7期1188-1196,共9页
Journal of Nuclear Agricultural Sciences
基金
科技部科技基础平台项目"核桃
板栗资源标准化整理
整合及共享试点"(2014-048)
农业部保种项目"核桃
板栗种质资源繁殖更新
鉴定评价与利用"(2014NWB009)
山东省农业良种工程项目(鲁科农字[2011]086)
中俄国际科技合作专项(2012DFR30700)
