干旱胁迫下割手密基因cDNA-SCoT差异表达分析
【目的】通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因。【方法】以割手密GX83-10为材料,构建正常浇灌(对照)及两个干旱处理叶片总RNA混合池,使用cDNA-SCoT构建割手密GX83-10伸长期应答干旱胁迫的基因表达谱,筛选、分离转录衍生片段(TDFs)并进行测序,根据NCBI数据库BLAST同源性检索结果推测基因功能,并使用qRT-PCR对抗旱相关TDFs进行表达验证分析。【结果】成功获得120个上调表达TDFs序列,其中53个TDFs序列与NCBI数据库中已录入的基因具有较高相似性,根据同源基因功能可分为10个类群:结合功能...
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| Published in | 南方农业学报 Vol. 49; no. 2; pp. 201 - 207 |
|---|---|
| Main Author | |
| Format | Journal Article |
| Language | Chinese |
| Published |
广西农业科学院 甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,南宁,530007%广西农业科学院 生物技术研究所,南宁,530007%广西作物遗传改良生物技术重点开放实验室,南宁,530007
2018
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| Subjects | |
| Online Access | Get full text |
| ISSN | 2095-1191 |
| DOI | 10.3969/j.issn.2095-1191.2018.02.01 |
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| Abstract | 【目的】通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因。【方法】以割手密GX83-10为材料,构建正常浇灌(对照)及两个干旱处理叶片总RNA混合池,使用cDNA-SCoT构建割手密GX83-10伸长期应答干旱胁迫的基因表达谱,筛选、分离转录衍生片段(TDFs)并进行测序,根据NCBI数据库BLAST同源性检索结果推测基因功能,并使用qRT-PCR对抗旱相关TDFs进行表达验证分析。【结果】成功获得120个上调表达TDFs序列,其中53个TDFs序列与NCBI数据库中已录入的基因具有较高相似性,根据同源基因功能可分为10个类群:结合功能蛋白相关基因(20.75%)、新陈代谢相关基因(13.21%)、通信及信号转导相关基因(13.21%)、转录调控因子相关基因(13.21%)、运输途径相关基因(11.32%)、环境互作相关基因(9.43%)、能量代谢相关基因(7.55%)、蛋白质合成相关基因(3.77%)、防御相关基因(3.77%)及细胞成分生物合成相关基因(3.77%)。通过对运输因子家族蛋白、RING/U-box超家族蛋白、22kD干旱诱导蛋白、微管蛋白alpha-3链和质膜H+-ATPase进行qRT-PCR分析,结果表明,这些基因在干旱胁迫下均呈不同程度的上调表达。【结论】干旱胁迫下,应用cDNA-SCoT构建割手密叶片基因表达谱筛选抗旱相关基因具有可行性。从割手密叶片中挖掘获得的抗旱及水分有效利用相关基因,可为利用野生种质资源开展甘蔗育种研究提供候选基因,改良甘蔗抗旱性,拓宽甘蔗育种基因库。 |
|---|---|
| AbstractList | S566.1; [目的]通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因.[方法]以割手密GX83-10为材料,构建正常浇灌(对照)及两个干旱处理叶片总RNA混合池,使用cDNA-SCoT构建割手密GX83-10伸长期应答干旱胁迫的基因表达谱,筛选、分离转录衍生片段(TDFs)并进行测序,根据NCBI数据库BLAST同源性检索结果推测基因功能,并使用qRT-PCR对抗旱相关TDFs进行表达验证分析.[结果]成功获得120个上调表达TDFs序列,其中53个TDFs序列与NCBI数据库中已录入的基因具有较高相似性,根据同源基因功能可分为10个类群:结合功能蛋白相关基因(20.75%)、新陈代谢相关基因(13.21%)、通信及信号转导相关基因(13.21%)、转录调控因子相关基因(13.21%)、运输途径相关基因(11.32%)、环境互作相关基因(9.43%)、能量代谢相关基因(7.55%)、蛋白质合成相关基因(3.77%)、防御相关基因(3.77%)及细胞成分生物合成相关基因(3.77%).通过对运输因子家族蛋白、RING/U-box超家族蛋白、22 kD干旱诱导蛋白、微管蛋白alpha-3链和质膜H+-ATPase进行qRT-PCR分析,结果表明,这些基因在干旱胁迫下均呈不同程度的上调表达.[结论]干旱胁迫下,应用cDNA-SCoT构建割手密叶片基因表达谱筛选抗旱相关基因具有可行性.从割手密叶片中挖掘获得的抗旱及水分有效利用相关基因,可为利用野生种质资源开展甘蔗育种研究提供候选基因,改良甘蔗抗旱性,拓宽甘蔗育种基因库. 【目的】通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因。【方法】以割手密GX83-10为材料,构建正常浇灌(对照)及两个干旱处理叶片总RNA混合池,使用cDNA-SCoT构建割手密GX83-10伸长期应答干旱胁迫的基因表达谱,筛选、分离转录衍生片段(TDFs)并进行测序,根据NCBI数据库BLAST同源性检索结果推测基因功能,并使用qRT-PCR对抗旱相关TDFs进行表达验证分析。【结果】成功获得120个上调表达TDFs序列,其中53个TDFs序列与NCBI数据库中已录入的基因具有较高相似性,根据同源基因功能可分为10个类群:结合功能蛋白相关基因(20.75%)、新陈代谢相关基因(13.21%)、通信及信号转导相关基因(13.21%)、转录调控因子相关基因(13.21%)、运输途径相关基因(11.32%)、环境互作相关基因(9.43%)、能量代谢相关基因(7.55%)、蛋白质合成相关基因(3.77%)、防御相关基因(3.77%)及细胞成分生物合成相关基因(3.77%)。通过对运输因子家族蛋白、RING/U-box超家族蛋白、22kD干旱诱导蛋白、微管蛋白alpha-3链和质膜H+-ATPase进行qRT-PCR分析,结果表明,这些基因在干旱胁迫下均呈不同程度的上调表达。【结论】干旱胁迫下,应用cDNA-SCoT构建割手密叶片基因表达谱筛选抗旱相关基因具有可行性。从割手密叶片中挖掘获得的抗旱及水分有效利用相关基因,可为利用野生种质资源开展甘蔗育种研究提供候选基因,改良甘蔗抗旱性,拓宽甘蔗育种基因库。 |
| Abstract_FL | [Objective]In order to provide candidate genes for improving drought resistance of sugarcane,the gene expression profiles in Saccharum spontaneum leaf under drought stress were analyzed for screening drought resistant genes.[Method]S. spontaneum GX83-10 was used as material to design two treatments viz. normal watering(control, CK)and drought stress(T),and their equal quantity mixed solutions of total RNA were constructed to establish gene ex-pression profiles in response to drought stress at elongation stage of GX83-10 by using cDNA-SCoT differential display technology.The transcript derived fragments(TDFs)were screened and isolated for sequencing,followed by the BLAST homology searching in NCBI database to deduce gene function based on homological genes. Furthermore,the related genes of drought resistance were analyzed by real time fluorescence quantitative PCR(qRT-PCR).[Result]In this study, 120 up-regulated TDFs were successfully cloned and sequenced.Among them,53 had higher similarity than others with the accessed genes in NCBI database,and these TDFs could be classified into 10 functional groups,including associated-functional protein related genes(20.75%),metabolism related genes(13.21%),communication and signal transduction related genes(13.21%),transcription regulated factor related genes(13.21%),transport pathway related genes(11.32%), environmental interaction related genes(9.43%),energy metabolism related genes(7.55%),protein synthesis related genes (3.77%),defense related genes(3.77%)and cell component biological synthesis related gene(3.77%).Transport factor 2 family protein,RING/U-box superfamily protein,22 kD drought-inducible protein,tubulin alpha-3 chain and plasma-membrane H+-ATPase were selected to conduct qRT-PCR analysis,and the results confirmed that all five genes were up-regulated in expressions at varying degrees under drought stress.[Conclusion]Under drought stress,it is feasible to screen the drought resistance related genes in S.spontaneum from gene expression profiles constructed by applying cDNA-SCoT differential display technology.The drought resistance and water efficient utilization genes,which were excavated from S. spontaneum leaf,will provide more candidate genes for sugarcane breeding research using wild germplasm re-sources to improve sugarcane drought resistance and broaden the genetic library of sugarcane breeding. |
| Author | 吴凯朝;韦莉萍;徐林;唐仕云;魏源文;黄诚梅;曹辉庆;罗海斌;蒋胜理;邓智年;李杨瑞 |
| AuthorAffiliation | 广西农业科学院甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,南宁530007;广西农业科学院生物技术研究所,南宁530007;广西作物遗传改良生物技术重点开放实验室,南宁530007 |
| AuthorAffiliation_xml | – name: 广西农业科学院 甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,南宁,530007%广西农业科学院 生物技术研究所,南宁,530007%广西作物遗传改良生物技术重点开放实验室,南宁,530007 |
| Author_FL | CAO Hui-qing LI Yang-rui HUANG Cheng-mei DENG Zhi-nian TANG Shi-yun XU Lin JIANG Sheng-li WU Kai-chao WEI Li-ping WEI Yuan-wen LUO Hai-bin |
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| DocumentTitleAlternate | Differentially expressed gene analysis by cDNA-SCoT in Saccharum spontaneum under drought stress |
| DocumentTitle_FL | Differentially expressed gene analysis by cDNA-SCoT in Saccharum spontaneum under drought stress |
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| Keywords | drought stress 干旱胁迫 transcript derived fragments(TDFs) gene cDNA-SCoT Saccharum spontaneum 转录衍生片段(TDFs) 割手密 基因 |
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| Notes | Saccharum spontaneum;cDNA-SCoT;transcript derived fragments(TDFs);gene;drought stress 45-1381/S Objective】In order to provide candidate genes for improving drought resistance of sugarcane,the gene expression profiles in Saccharum spontaneum leaf under drought stress were analyzed for screening drought resistant genes【Method】S.spontaneum GX83-10was used as material to design two treatments viz.normal watering(control,CK)and drought stress(T),and their equal quantity mixed solutions of total RNA were constructed to establish gene expression profiles in response to drought stress at elongation stage of GX83-10by using cDNA-SCoT differential display technology.The transcript derived fragments(TDFs)were screened and isolated for sequencing,followed by the BLAST homology searching in NCBI database to deduce gene function based on homological genes.Furthermore,the related genes of drought resistance were analyzed by real time fluorescence quantitative PCR(qRT-PCR).【Result】In this study,120up-regulated TDFs were succes |
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| PublicationTitle | 南方农业学报 |
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| PublicationYear | 2018 |
| Publisher | 广西农业科学院 甘蔗研究所/农业部广西甘蔗生物技术与遗传改良重点实验室/广西甘蔗遗传改良重点实验室,南宁,530007%广西农业科学院 生物技术研究所,南宁,530007%广西作物遗传改良生物技术重点开放实验室,南宁,530007 |
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| Snippet | 【目的】通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因。【方法】以割手密GX83-10为材料,构建正常浇灌(对照)及... S566.1; [目的]通过分析割手密叶片在干旱胁迫下的基因表达谱,筛选获得抗旱相关基因,为开展甘蔗抗旱性遗传改良研究提供候选基因.[方法]以割手密GX83-10为材料,构建正常浇... |
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| SubjectTerms | 割手密;cDNA-SCoT;转录衍生片段(TDFs);基因;干旱胁迫 |
| Title | 干旱胁迫下割手密基因cDNA-SCoT差异表达分析 |
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