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葡萄SCARECROW基因家族的分析与表达
陈丽华1, 葛晖1, 柴丽娟1, 陈尚武2, 马会勤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.中国农业大学 食品科学与营养工程学院,北京 100083)
摘要:
赤霉素(GA)促进葡萄果实的膨大,目前对外源赤霉素促进葡萄果实膨大的信号转导途径还不完全清楚。有研究发现GA3处理后葡萄果实中一个SCARECROW(SCR)蛋白的表达出现上调,为了进一步揭示GA的作用途径,本研究通过分析在其他植物中已经确认的SCR基因序列,对葡萄全基因组进行BLAST比对,获得了可能编码葡萄SCR的11个基因片段序列SCRs。分别设计特异性引物,以无核白鸡心葡萄(Vitis vinifera L.cv.Centennial seedless)不同组织器官,以及盛花后12 d以30 mg/L GA3处理无核白鸡心葡萄的花序,并于花后13、15、19、45、61 d采收的果实为材料,进行半定量RT-PCR测定SCRs的表达。结果显示:这些基因在成叶、幼叶、根、花序、新梢顶尖、休眠芽、胚性愈伤组织和非胚性愈伤组织中存在差异表达;在花后13、19、45、61 d的葡萄果实中有不同状态的上调。研究结果对进一步探讨SCRs在葡萄果实发育和成熟过程的作用与功能提供了线索。
关键词:  葡萄  SCARECROW  GA3处理  半定量  基因家族
DOI:10.11841/j.issn.1007-4333.2012.01.013
投稿时间:2011-07-05
基金项目:中国农业大学基本科研业务费资助项目(2009JS51)
Putative SCARECROW gene family of Vitis vinifera L. andtheir differential expression under GA treatment
CHEN Li-hua1, GE Hui1, CHAI Li-juan1, CHEN Shang-wu2, MA Hui-qin1
(1.College of Agronomy and Biotechnology,China Agricultural University,Beijing 100193,China;2.College of Food Science and Nutritional Engineering,China Agricultural University,Beijing 100083,China)
Abstract:
Gibberellins (GA) are important plant hormone which promotes the enlargement of seedless grape berries.GA signal transduction,especially downstream elements,in the process of grape berry enlargement is still poorly known.Our previous proteomic results demonstrated that a SCARECROW(SCR) protein was up-regulated in GA3 treated seedless grape berries.As a mining step,11 sequences were identified from the grape genome as putative SCR gene,based on their coding protein's similarity to SCR identified in other plants.Specific primers were designed after BLAST analysis.Bunches of Centennial seedless were treated with 30mg/L GA3 on 12 days after flowering (D).The fruit were collected at the first rapid (13,15 and 19 d),slow (45 d) and the second rapid growth stage (61 d).Semi-quantitative RT-PCR showed that these 11 genes were of different transcription levels in mature leaves,young leaves,inflorescences,root tips,shoot tips,dormant buds,embryonic callus (EC) and non-embryonic callus (NEC).And these putative SCR genes,were up-regulated on 13,19,45,61 d by GA3 treatment.Our results lay the ground for future work on full sequence clone and functional study of grape SCR genes.
Key words:  grape  SCARECROW  GA3  semi-quantitative RT-PCR  gene family