本文主要研究内容
作者王少波（2019）在《水稻耐热胁迫基因的挖掘及其表达分析》一文中研究指出：我国是世界上最大的水稻生产国和消费国,全球有超过一半的人口以水稻为主要粮食。水稻(Oryza sativa L.)属喜温作物,其生长需要适宜的温度。当环境温度超出水稻适宜生长温度的上限时,会影响其正常的生长发育并对产量造成损失。培育耐高温胁迫的水稻新品种是应对全球气候变暖导致气温升高的有效策略。随着水稻3K基因组测序工作的完成,使得从大量水稻品种的基因组中挖掘出耐热基因的研究成为可能,这可为耐高温胁迫水稻新品种的选育积累宝贵的基因资源。本研究首先从水稻3K基因组计划和NCBI数据库中下载获取22个具有不同耐热胁迫能力水稻品种的基因组重测序数据。其中水稻品种Azucena(CX151)为高温敏感型品种,其余21个为耐热胁迫型水稻品种;也从NCBI数据库的GEO数据库中下载热胁迫处理下高温敏感型水稻品种Azucena的RNA-seq数据,及前人研究获得的热胁迫处理下的耐热胁迫型水稻品种CO39的差异表达基因数据。然后,以The Rice Annotation Project(RAP)下载的Nipponbare基因组(Ver.7.0)为参考基因组,通过比较基因组学和比较转录组学系统的开展水稻抗热胁迫基因的挖掘,并分析其表达模式,主要研究结果如下所述。1.从基因组重测序数据中挖掘获得水稻耐热胁迫基因本文以水稻品种Nipponbare的基因组序列为参考基因组,将CO39、Pokkili和Bala等21种耐热胁迫型水稻品种和高温敏感型品种Azucena的基因组重测序数据分别比对到参考基因组上,再分别对耐热胁迫型水稻和高温敏感型水稻进行单核苷酸多态性(Single Nucleotide Polymorphisms,SNP)位点检测,其中在Azucena中共检测到920,481个高质量的SNPs,在21种耐热胁迫型水稻中共检测到了4,181,396个特异于高温敏感型水稻Azucena的高质量SNPs。通过对SNPs位点的注释,筛选分析其中的非同义突变(non-synonymous SNPs,nsSNPs)和无义突变(nonsense SNPs),该类型的变异能引起基因所编码蛋白质的变化。最终,筛选得到525个在21个耐热胁迫型水稻品种中均存在的SNPs,其中有20个为nsSNPs,这些nsSNPs映射到15个基因中,分别为Os11g0587000,Os04g0244400,Os07g0614100,Os10g0168751,Os11g0233000,Os12g0225700,Os01g0153800,Os12g0438100,Os01g0150800,Os12g0631800,Os11g0229700,Os08g0254600,Os10g0133800,Os09g0349700和Os10g0141300。为下一步开展水稻潜在耐热基因的功能验证试验指明了方向。2.从转录组测序数据中挖掘获得水稻受热胁迫诱导表达基因从NCBI的GEO数据库下载热胁迫处理下高温敏感型水稻品种Azucena的RNA-seq数据,应用RNA-seq分析技术,获得热胁迫下水稻品种Azucena的差异表达基因。结果发现在热胁迫处理15min时,相对于其他时期,水稻品种Azucena中差异表达基因的数量最多,共检测到2486个差异表达基因,其中上调差异表达基因为1480个,下调差异表达基因为1006个。说明高温胁迫15min是水稻品种Azucena应对胁迫的关键时期。同时,本文也获得了在高温胁迫处理下耐热胁迫型水稻CO39中差异表达转录本,共3487个差异表达转录本,上调差异表达转录本有1014个,下调差异表达转录本有1498个。然后,筛选耐热胁迫型水稻品种CO39中特异于高温敏感型水稻品种Azucena的差异表达基因有2262个,表明耐热能力不同的水稻品种对热胁迫应激反应的模式不同。GO注释结果表明,两者相同的差异表达基因主要参与细胞稳态过程和蛋白质折叠,表明热胁迫下保持水稻内环境稳态是维持水稻生长最基本的条件。3.获得3个具有耐热nsSNPs又受热诱导表达的水稻基因本研究发现,在耐热水稻品种CO39中受热胁迫诱导上调表达的3个基因Os09g0349700、Os11g0229700和Os12g0631800,都有耐热胁迫特有的nsSNPs位点,预测分析发现nsSNPs分别能影响该3个基因蛋白质的功能,表明位点突变和受热胁迫诱导差异表达,使这3个基因在水稻应对热胁迫中发挥重要作用。其中Os09g0349700基因影响水稻的水分利用效率,而其存在的一个nsSNP使编码蛋白质序列中的第96位氨基酸由天冬酰胺突变成苏氨酸,导致耐热水稻品种获得耐高温胁迫能力。Os11g0229700基因功能与NBS-LRR蛋白类似,也是一种抗逆基因。热胁迫条件下耐热水稻品种CO39中Os12g0631800基因被上调表达5.06倍,且该基因在21种耐热胁迫型水稻中均存在一个nsSNP,使编码该蛋白的第143位氨基酸由蛋氨酸突变成苏氨酸,可能参与植株体内的氧化还原反应,促进植株耐高温胁迫。本文以基因组重测序数据和转录组数据为基础,通过比较高温敏感型水稻品种Azucena和21种耐热胁迫型水稻品种间的nsSNPs,挖掘到水稻潜在的耐热基因;也通过比较转录组学技术分析高温胁迫下耐热水稻品种CO39和高温敏感型水稻品种Azucena间不同的差异表达基因,挖掘到受热诱导上调表达的基因;最后,还筛选到3个既受热诱导上调表达又具有耐热nsSNPs的基因,为后续的基因功能验证试验指明了方向。
Abstract
wo guo shi shi jie shang zui da de shui dao sheng chan guo he xiao fei guo ,quan qiu you chao guo yi ban de ren kou yi shui dao wei zhu yao liang shi 。shui dao (Oryza sativa L.)shu xi wen zuo wu ,ji sheng chang xu yao kuo yi de wen du 。dang huan jing wen du chao chu shui dao kuo yi sheng chang wen du de shang xian shi ,hui ying xiang ji zheng chang de sheng chang fa yo bing dui chan liang zao cheng sun shi 。pei yo nai gao wen xie pai de shui dao xin pin chong shi ying dui quan qiu qi hou bian nuan dao zhi qi wen sheng gao de you xiao ce lve 。sui zhao shui dao 3Kji yin zu ce xu gong zuo de wan cheng ,shi de cong da liang shui dao pin chong de ji yin zu zhong wa jue chu nai re ji yin de yan jiu cheng wei ke neng ,zhe ke wei nai gao wen xie pai shui dao xin pin chong de shua yo ji lei bao gui de ji yin zi yuan 。ben yan jiu shou xian cong shui dao 3Kji yin zu ji hua he NCBIshu ju ku zhong xia zai huo qu 22ge ju you bu tong nai re xie pai neng li shui dao pin chong de ji yin zu chong ce xu shu ju 。ji zhong shui dao pin chong Azucena(CX151)wei gao wen min gan xing pin chong ,ji yu 21ge wei nai re xie pai xing shui dao pin chong ;ye cong NCBIshu ju ku de GEOshu ju ku zhong xia zai re xie pai chu li xia gao wen min gan xing shui dao pin chong Azucenade RNA-seqshu ju ,ji qian ren yan jiu huo de de re xie pai chu li xia de nai re xie pai xing shui dao pin chong CO39de cha yi biao da ji yin shu ju 。ran hou ,yi The Rice Annotation Project(RAP)xia zai de Nipponbareji yin zu (Ver.7.0)wei can kao ji yin zu ,tong guo bi jiao ji yin zu xue he bi jiao zhuai lu zu xue ji tong de kai zhan shui dao kang re xie pai ji yin de wa jue ,bing fen xi ji biao da mo shi ,zhu yao yan jiu jie guo ru xia suo shu 。1.cong ji yin zu chong ce xu shu ju zhong wa jue huo de shui dao nai re xie pai ji yin ben wen yi shui dao pin chong Nipponbarede ji yin zu xu lie wei can kao ji yin zu ,jiang CO39、Pokkilihe Baladeng 21chong nai re xie pai xing shui dao pin chong he gao wen min gan xing pin chong Azucenade ji yin zu chong ce xu shu ju fen bie bi dui dao can kao ji yin zu shang ,zai fen bie dui nai re xie pai xing shui dao he gao wen min gan xing shui dao jin hang chan he gan suan duo tai xing (Single Nucleotide Polymorphisms,SNP)wei dian jian ce ,ji zhong zai Azucenazhong gong jian ce dao 920,481ge gao zhi liang de SNPs,zai 21chong nai re xie pai xing shui dao zhong gong jian ce dao le 4,181,396ge te yi yu gao wen min gan xing shui dao Azucenade gao zhi liang SNPs。tong guo dui SNPswei dian de zhu shi ,shai shua fen xi ji zhong de fei tong yi tu bian (non-synonymous SNPs,nsSNPs)he mo yi tu bian (nonsense SNPs),gai lei xing de bian yi neng yin qi ji yin suo bian ma dan bai zhi de bian hua 。zui zhong ,shai shua de dao 525ge zai 21ge nai re xie pai xing shui dao pin chong zhong jun cun zai de SNPs,ji zhong you 20ge wei nsSNPs,zhe xie nsSNPsying she dao 15ge ji yin zhong ,fen bie wei Os11g0587000,Os04g0244400,Os07g0614100,Os10g0168751,Os11g0233000,Os12g0225700,Os01g0153800,Os12g0438100,Os01g0150800,Os12g0631800,Os11g0229700,Os08g0254600,Os10g0133800,Os09g0349700he Os10g0141300。wei xia yi bu kai zhan shui dao qian zai nai re ji yin de gong neng yan zheng shi yan zhi ming le fang xiang 。2.cong zhuai lu zu ce xu shu ju zhong wa jue huo de shui dao shou re xie pai you dao biao da ji yin cong NCBIde GEOshu ju ku xia zai re xie pai chu li xia gao wen min gan xing shui dao pin chong Azucenade RNA-seqshu ju ,ying yong RNA-seqfen xi ji shu ,huo de re xie pai xia shui dao pin chong Azucenade cha yi biao da ji yin 。jie guo fa xian zai re xie pai chu li 15minshi ,xiang dui yu ji ta shi ji ,shui dao pin chong Azucenazhong cha yi biao da ji yin de shu liang zui duo ,gong jian ce dao 2486ge cha yi biao da ji yin ,ji zhong shang diao cha yi biao da ji yin wei 1480ge ,xia diao cha yi biao da ji yin wei 1006ge 。shui ming gao wen xie pai 15minshi shui dao pin chong Azucenaying dui xie pai de guan jian shi ji 。tong shi ,ben wen ye huo de le zai gao wen xie pai chu li xia nai re xie pai xing shui dao CO39zhong cha yi biao da zhuai lu ben ,gong 3487ge cha yi biao da zhuai lu ben ,shang diao cha yi biao da zhuai lu ben you 1014ge ,xia diao cha yi biao da zhuai lu ben you 1498ge 。ran hou ,shai shua nai re xie pai xing shui dao pin chong CO39zhong te yi yu gao wen min gan xing shui dao pin chong Azucenade cha yi biao da ji yin you 2262ge ,biao ming nai re neng li bu tong de shui dao pin chong dui re xie pai ying ji fan ying de mo shi bu tong 。GOzhu shi jie guo biao ming ,liang zhe xiang tong de cha yi biao da ji yin zhu yao can yu xi bao wen tai guo cheng he dan bai zhi she die ,biao ming re xie pai xia bao chi shui dao nei huan jing wen tai shi wei chi shui dao sheng chang zui ji ben de tiao jian 。3.huo de 3ge ju you nai re nsSNPsyou shou re you dao biao da de shui dao ji yin ben yan jiu fa xian ,zai nai re shui dao pin chong CO39zhong shou re xie pai you dao shang diao biao da de 3ge ji yin Os09g0349700、Os11g0229700he Os12g0631800,dou you nai re xie pai te you de nsSNPswei dian ,yu ce fen xi fa xian nsSNPsfen bie neng ying xiang gai 3ge ji yin dan bai zhi de gong neng ,biao ming wei dian tu bian he shou re xie pai you dao cha yi biao da ,shi zhe 3ge ji yin zai shui dao ying dui re xie pai zhong fa hui chong yao zuo yong 。ji zhong Os09g0349700ji yin ying xiang shui dao de shui fen li yong xiao lv ,er ji cun zai de yi ge nsSNPshi bian ma dan bai zhi xu lie zhong de di 96wei an ji suan you tian dong xian an tu bian cheng su an suan ,dao zhi nai re shui dao pin chong huo de nai gao wen xie pai neng li 。Os11g0229700ji yin gong neng yu NBS-LRRdan bai lei shi ,ye shi yi chong kang ni ji yin 。re xie pai tiao jian xia nai re shui dao pin chong CO39zhong Os12g0631800ji yin bei shang diao biao da 5.06bei ,ju gai ji yin zai 21chong nai re xie pai xing shui dao zhong jun cun zai yi ge nsSNP,shi bian ma gai dan bai de di 143wei an ji suan you dan an suan tu bian cheng su an suan ,ke neng can yu zhi zhu ti nei de yang hua hai yuan fan ying ,cu jin zhi zhu nai gao wen xie pai 。ben wen yi ji yin zu chong ce xu shu ju he zhuai lu zu shu ju wei ji chu ,tong guo bi jiao gao wen min gan xing shui dao pin chong Azucenahe 21chong nai re xie pai xing shui dao pin chong jian de nsSNPs,wa jue dao shui dao qian zai de nai re ji yin ;ye tong guo bi jiao zhuai lu zu xue ji shu fen xi gao wen xie pai xia nai re shui dao pin chong CO39he gao wen min gan xing shui dao pin chong Azucenajian bu tong de cha yi biao da ji yin ,wa jue dao shou re you dao shang diao biao da de ji yin ;zui hou ,hai shai shua dao 3ge ji shou re you dao shang diao biao da you ju you nai re nsSNPsde ji yin ,wei hou xu de ji yin gong neng yan zheng shi yan zhi ming le fang xiang 。
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论文作者分别是来自福建农林大学的王少波，发表于刊物福建农林大学2019-08-30论文，是一篇关于水稻论文,高温胁迫论文,功能基因挖掘论文,基因组重测序论文,转录组论文,福建农林大学2019-08-30论文的文章。本文可供学术参考使用，各位学者可以免费参考阅读下载，文章观点不代表本站观点，资料来自福建农林大学2019-08-30论文网站，若本站收录的文献无意侵犯了您的著作版权，请联系我们删除。
标签：水稻论文; 高温胁迫论文; 功能基因挖掘论文; 基因组重测序论文; 转录组论文; 福建农林大学2019-08-30论文;