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苹果腺苷-异戊烯基转移酶基因的组织表达分析
任雪芹1, 许轲1, 李皓2, 张文1, 朱元娣1
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(1.中国农业大学 农学与生物技术学院, 北京 100193;2.国家知识产权局专利局 专利审查协作北京中心, 北京 100010)
摘要:
为深入研究MdIPTs基因在苹果细胞分裂素合成途径中的作用和利用转基因技术改良苹果树型,分析苹果(Malus×Domestica Borkh.) A-IPTs基因的时空表达,本研究以 '富士’苹果(M.Domestica Borkh.cv.Fuji)及其组培苗为试材,利用半定量RT-PCR方法分析7个A-IPTs基因在梢尖、幼叶、茎段节间、皮部组织、叶芽、花芽、花蕾、花、幼果和根部的表达水平。结果表明:MdIPT3a总体表达水平最高、MdIPT1b次之,在所检测的器官组织中均有表达;MdIPT5a在花蕾和花中表达量高,MdIPT5b在梢尖和幼叶中表达量高;MdIPT1a在根、皮部组织和叶芽中检测到微弱表达,MdIPT3b仅在组培苗的茎段中节间微弱表达;所检测的组织器官中未检测到MdIPT7a表达。苹果7个A-IPTs基因具有组织表达特异性,高度同源的成对基因的组织表达模式不同。
关键词:  苹果  细胞分裂素  腺苷-异戊烯基转移酶  半定量RT-PCR  表达分析
DOI:10.11841/j.issn.1007-4333.2013.06.016
投稿时间:2013-04-01
基金项目:国家自然科学基金项目(31070614)
Expression analysis of the adenylate-isopentenyltransferase(A-IPT) genes in apple(Malus×Domestica Borkh.)
REN Xue-qin1, XU Ke1, LI Hao2, ZHANG Wen1, ZHU Yuan-di1
(1.College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China;2.Patent Examination Cooperation Center of the Patent Office, SIPO, Beijing 100010, China)
Abstract:
Tissue-specific expression of the A-IPTs genes in apple(Malus×Domestica Borkh.) were analyzed for further study of roles of MdIPTs in cytokinin biosynthesis and modifying tree form by genetic transformation in apple.In this study,the ‘Fuji’ apple(M.domestica Borkh.cv.Fuji) and its plantlets in vitro were used as materials and semi-quantitative reverse transcription-polymerase chain reaction(RT-PCR) was conducted to study expression levels of seven A-IPTs in shoot tips,young leaves,internodes,bark tissues,leaf buds,flora buds,flowers at balloon stage,open flowers,young fruits and roots of ‘Fuji’ apple.The results showed that MdIPT3a and MdIPT1b were detected in all tested tissues and organs.The expression lever of MdIPT3a was higher than that of MdIPT1b.MdIPT5a expressed highly in flowers,while MdIPT5b in shoot tips and young leaves.The weak expression of MdIPT1a was detected in roots,bark tissue,and leaf buds,while MdIPT3b was only detected in internodes of plantlets in vitro.MdIPT7a wasnt detected in all tested tissues.Seven A-IPTs exhibit tissue-specific expression and highly homologous genes in pairs in apple had different expression profiles.
Key words:  Malus×Domestica Borkh.  cytokinins  adenylate-isopentenyltransferase  semi-quantitative RT-PCR  expression analysis