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基于cDNA-AFLP发掘茶树被茶尺蠖取食诱导的相关差异基因及其表达特征
  • ISSN号:0578-1752
  • 期刊名称:《中国农业科学》
  • 时间:0
  • 分类:S513[农业科学—作物学]
  • 作者机构:[1]安徽农业大学茶与食品科技学院/茶叶生物化学与生物技术省部共建国家重点实验室培育基地/农业部茶树生物学与茶叶加工重点实验室,合肥230036
  • 相关基金:国家自然科学基金项目(306713312)、“十二五”科技支撑计划子课题(2011BAD01B01)、长江学者和创新团队发展计划(IRT1101)、安徽省重点实验室科技计划(10111406022)
中文摘要:

【目的】发掘茶树被茶尺蠖取食诱导的相关防御基因及其分子防御机制。【方法】利用cDNA-AFLP筛选茶树叶片被其主要害虫茶尺蠖取食后差异表达片段(transcript-derived fragment,TDF),并对其进行生物信息学分析和相关验证;运用qRT-PCR(quantitative real-time PCR)研究部分差异基因的表达规律。【结果】茶树被茶尺蠖取食后,共发现231条TDF,经PCR验证和测序后,成功获得134条EST(expressed sequence tags)序列,其中上调与下调表达的TDF数量分别为81和53条,各占测序获得序列的60.4%与39.6%;利用BLASTx在GenBank进行比对与功能分析可知,其中的23.1%与代谢相关,9.0%跟光合作用与能量相关,跟蛋白质合成与贮藏相关的序列各占8.2%,与信号转导相关的占6.0%,跟抗病与防御相关的占5.2%,但与细胞生长及细胞结构、转录因子和运输相关的数量很少,分别为1.5%、2.2%和3.0%;此外,还有6.7%的TDF功能不明确,有20.1%的无同源性序列;利用qRT-PCR(quantitative real-time PCR)对TDF 21F1、22D、27L3和28F分别进行了表达特征分析,结果表明这4个TDF代表的基因在茶树被茶尺蠖取食后均有不同程度诱导。【结论】通过cDNA-AFLP技术发掘了多个茶树被茶尺蠖取食诱导的差异表达基因片段,初步揭示了茶树被害虫取食而诱导的分子防御机制。

英文摘要:

【Objective】 The objective of this study is to screen the defense genes in tea plant (Camellia sinensis L.) induced by tea looper (Ectropis oblique).【Method】Tea leaves fed by the looper were analyzed by cDNA-AFLP(cDNA-amplified fragment length polymorphism). Transcript-derived fragments (TDF) were screened and confirmed. After that, bioinformatics analysis was followed. And the expression patterns of the differential genes were performed by qRT-PCR.【Result】The results showed that 134 EST(expressed sequence tags) were obtained by sequencing and identifying from the selected 231 TDFs. Among them, 81 TDFs were up-regulated, and 53 were down-regulated, which accounted for 60.4% and 39.6% of the obtained fragments, respectively. According to BLASTx result of TDFs, a large group (23.1%) of these genes shared high homology with genes involved in metabolism, 9.0% genes with functions related to photosynthesis and energy, 8.2% genes with functions related to protein synthesis and storage, signal transduction, 5.2% genes with functions related to disease/defense. But only few genes involved in cell growth and structure, transcription factor and transporters and the ratios were 1.5%, 2.2%, and 3.0%, respectively. Moreover, sequences with unknown proteins occupied 6.7%; and 20.1% sequences with no similarity against the database. TDF-21F1, TDF-22D, TDF-27L3 and TDF-28F were confirmed by qRT-PCR analysis, further result indicated that the expression of the defense genes responsive to Ectropis oblique play significant regulation roles in adapting to negative environment for tea plant.【Conclusion】Using cDNA-AFLP method to analyze Ectropis oblique feeding tea plant and get the TDFs, it could supply valuable information to the study of the molecular mechanism between tea and pest.

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期刊信息
  • 《中国农业科学》
  • 中国科技核心期刊
  • 主管单位:中华人民共和国农业部
  • 主办单位:中国农业科学院 中国农学会
  • 主编:万建民
  • 地址:北京中关村南大街12号中国农业科学院图书馆楼4101-4103室
  • 邮编:100081
  • 邮箱:zgnykx@caas.cn
  • 电话:010-82109808 82106279
  • 国际标准刊号:ISSN:0578-1752
  • 国内统一刊号:ISSN:11-1328/S
  • 邮发代号:2-138
  • 获奖情况:
  • 中国期刊方阵“双高”期刊,第三届中国出版政府奖提名奖
  • 国内外数据库收录:
  • 美国化学文摘(网络版),英国农业与生物科学研究中心文摘,波兰哥白尼索引,英国动物学记录,日本日本科学技术振兴机构数据库,中国中国科技核心期刊,中国北大核心期刊(2004版),中国北大核心期刊(2008版),中国北大核心期刊(2011版),中国北大核心期刊(2014版),英国食品科技文摘,中国北大核心期刊(2000版)
  • 被引量:85620