本文主要研究内容
作者季家磊（2019）在《甘蓝83121A隐性核雄性不育基因的克隆与功能分析》一文中研究指出：甘蓝(Brassica oleracea L.var.capitata L.)是一种重要的两年生蔬菜,属于十字花科芸薹属甘蓝种。甘蓝在产量、品质、抗病性、抗逆性等性状上具有明显的杂种优势。杂种优势育种已成为甘蓝品种改良的主要方式。雄性不育突变体在甘蓝杂种优势利用中发挥着重要作用,同时也是研究植物生殖发育调控的重要材料。83121A雄性不育材料是本课题组在育种过程中发现的隐性单核基因突变体。本研究以83121A为材料,研究了雄性不育发生的时期及细胞学特征,对雄性不育基因进行了克隆与功能验证,同时利用转录组和蛋白组技术探讨了雄性不育发生的分子机理,为该突变体材料在甘蓝育种中的应用奠定了基础。主要研究结果如下:1.83121A营养生长正常,花朵可以正常开放,蜜腺发育正常,花蜜较多,但花药败育彻底,无花粉。通过石蜡切片观察发现,83121A可正常形成四分体,四分小孢子释放后,绒毡层提前降解,小孢子的发育停滞在单核期并最终降解;另外,利用扫描电镜和透射电镜观察发现,不育材料的绒毡层液泡化严重,几乎检测不到绒毡层小体的存在,且83121A小孢子从四分体释放后无法形成花粉外壁,推测花粉外壁的缺失是小孢子败育的主要原因。2.利用转录组测序技术分析了不育系83121A及其近等基因可育系83121B小孢子双核期之前花蕾的基因表达,共得到2881个差异表达基因,其中在83121A中上调表达的有1310个,下调表达的有1571个。通过分析花粉外壁合成相关基因在83121AB中的表达,将与拟南芥CYP704B1同源的基因Bol023932确定为重要候选基因,并命名为BoCYP704B1,该基因在83121A花蕾中的表达受到严重抑制。3.BoCYP704B1在野生型甘蓝花药及绒毡层中特异表达,其表达量在四分体时期和单核早期达到最大,以后逐渐减弱至终止;亚细胞定位结果表明该蛋白定位在内质网中。BoCYP704B1与拟南芥CYP704B1、水稻CYP704B2同源性较高,目前已知CYP704B1和CYP704B2均催化中长链脂肪酸的羟基化,据此推测,BoCYP704B1可能编码脂肪酸羟基化酶,参与花粉外壁合成。4.通过克隆测序发现,83121A中BoCYP704B1基因的第一个外显子中插入一段5424bp的Ty3-gypsy类型反转录转座子;转座子的插入一方面抑制了该基因的表达,另一方面改变了转录剪切位点,使编码蛋白结构发生变化,丧失生物学功能。5.根据BoCYP704B1突变设计基因内分子标记进行连锁分析发现,纯合BoCYP704B1突变总是与雄性不育性状连锁,说明该基因突变是导致雄性不育的根本原因,开发的分子标记可以用于不育材料的辅助选择。另外通过转基因互补试验证明了BoCYP704B1可以恢复83121A的育性,是隐性核不育系83121A的恢复基因,可利用BoCYP704B1通过基因工程技术创制雄性不育保持系。6.采用高通量非标记定量蛋白质组学label-free的方法对83121A和83121B小孢子双核期之前的花蕾进行差异蛋白质组学分析,鉴定出1245个差异表达蛋白质,其中648个蛋白在83121A中下调表达。这些差异蛋白的生物学功能主要涉及花粉壁合成,脂肪酸代谢,氨基酸合成以及蛋白质加工修饰,表明这些代谢途径对于甘蓝生殖发育具有重要的作用。
Abstract
gan lan (Brassica oleracea L.var.capitata L.)shi yi chong chong yao de liang nian sheng shu cai ,shu yu shi zi hua ke yun tai shu gan lan chong 。gan lan zai chan liang 、pin zhi 、kang bing xing 、kang ni xing deng xing zhuang shang ju you ming xian de za chong you shi 。za chong you shi yo chong yi cheng wei gan lan pin chong gai liang de zhu yao fang shi 。xiong xing bu yo tu bian ti zai gan lan za chong you shi li yong zhong fa hui zhao chong yao zuo yong ,tong shi ye shi yan jiu zhi wu sheng shi fa yo diao kong de chong yao cai liao 。83121Axiong xing bu yo cai liao shi ben ke ti zu zai yo chong guo cheng zhong fa xian de yin xing chan he ji yin tu bian ti 。ben yan jiu yi 83121Awei cai liao ,yan jiu le xiong xing bu yo fa sheng de shi ji ji xi bao xue te zheng ,dui xiong xing bu yo ji yin jin hang le ke long yu gong neng yan zheng ,tong shi li yong zhuai lu zu he dan bai zu ji shu tan tao le xiong xing bu yo fa sheng de fen zi ji li ,wei gai tu bian ti cai liao zai gan lan yo chong zhong de ying yong dian ding le ji chu 。zhu yao yan jiu jie guo ru xia :1.83121Aying yang sheng chang zheng chang ,hua duo ke yi zheng chang kai fang ,mi xian fa yo zheng chang ,hua mi jiao duo ,dan hua yao bai yo che de ,mo hua fen 。tong guo dan la qie pian guan cha fa xian ,83121Ake zheng chang xing cheng si fen ti ,si fen xiao bao zi shi fang hou ,rong zhan ceng di qian jiang jie ,xiao bao zi de fa yo ting zhi zai chan he ji bing zui zhong jiang jie ;ling wai ,li yong sao miao dian jing he tou she dian jing guan cha fa xian ,bu yo cai liao de rong zhan ceng ye pao hua yan chong ,ji hu jian ce bu dao rong zhan ceng xiao ti de cun zai ,ju 83121Axiao bao zi cong si fen ti shi fang hou mo fa xing cheng hua fen wai bi ,tui ce hua fen wai bi de que shi shi xiao bao zi bai yo de zhu yao yuan yin 。2.li yong zhuai lu zu ce xu ji shu fen xi le bu yo ji 83121Aji ji jin deng ji yin ke yo ji 83121Bxiao bao zi shuang he ji zhi qian hua lei de ji yin biao da ,gong de dao 2881ge cha yi biao da ji yin ,ji zhong zai 83121Azhong shang diao biao da de you 1310ge ,xia diao biao da de you 1571ge 。tong guo fen xi hua fen wai bi ge cheng xiang guan ji yin zai 83121ABzhong de biao da ,jiang yu ni na gai CYP704B1tong yuan de ji yin Bol023932que ding wei chong yao hou shua ji yin ,bing ming ming wei BoCYP704B1,gai ji yin zai 83121Ahua lei zhong de biao da shou dao yan chong yi zhi 。3.BoCYP704B1zai ye sheng xing gan lan hua yao ji rong zhan ceng zhong te yi biao da ,ji biao da liang zai si fen ti shi ji he chan he zao ji da dao zui da ,yi hou zhu jian jian ruo zhi zhong zhi ;ya xi bao ding wei jie guo biao ming gai dan bai ding wei zai nei zhi wang zhong 。BoCYP704B1yu ni na gai CYP704B1、shui dao CYP704B2tong yuan xing jiao gao ,mu qian yi zhi CYP704B1he CYP704B2jun cui hua zhong chang lian zhi fang suan de qiang ji hua ,ju ci tui ce ,BoCYP704B1ke neng bian ma zhi fang suan qiang ji hua mei ,can yu hua fen wai bi ge cheng 。4.tong guo ke long ce xu fa xian ,83121Azhong BoCYP704B1ji yin de di yi ge wai xian zi zhong cha ru yi duan 5424bpde Ty3-gypsylei xing fan zhuai lu zhuai zuo zi ;zhuai zuo zi de cha ru yi fang mian yi zhi le gai ji yin de biao da ,ling yi fang mian gai bian le zhuai lu jian qie wei dian ,shi bian ma dan bai jie gou fa sheng bian hua ,sang shi sheng wu xue gong neng 。5.gen ju BoCYP704B1tu bian she ji ji yin nei fen zi biao ji jin hang lian suo fen xi fa xian ,chun ge BoCYP704B1tu bian zong shi yu xiong xing bu yo xing zhuang lian suo ,shui ming gai ji yin tu bian shi dao zhi xiong xing bu yo de gen ben yuan yin ,kai fa de fen zi biao ji ke yi yong yu bu yo cai liao de fu zhu shua ze 。ling wai tong guo zhuai ji yin hu bu shi yan zheng ming le BoCYP704B1ke yi hui fu 83121Ade yo xing ,shi yin xing he bu yo ji 83121Ade hui fu ji yin ,ke li yong BoCYP704B1tong guo ji yin gong cheng ji shu chuang zhi xiong xing bu yo bao chi ji 。6.cai yong gao tong liang fei biao ji ding liang dan bai zhi zu xue label-freede fang fa dui 83121Ahe 83121Bxiao bao zi shuang he ji zhi qian de hua lei jin hang cha yi dan bai zhi zu xue fen xi ,jian ding chu 1245ge cha yi biao da dan bai zhi ,ji zhong 648ge dan bai zai 83121Azhong xia diao biao da 。zhe xie cha yi dan bai de sheng wu xue gong neng zhu yao she ji hua fen bi ge cheng ,zhi fang suan dai xie ,an ji suan ge cheng yi ji dan bai zhi jia gong xiu shi ,biao ming zhe xie dai xie tu jing dui yu gan lan sheng shi fa yo ju you chong yao de zuo yong 。
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论文详细介绍
论文作者分别是来自中国农业科学院的季家磊，发表于刊物中国农业科学院2019-07-05论文，是一篇关于甘蓝论文,雄性不育论文,转录组论文,蛋白组论文,中国农业科学院2019-07-05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，资料来自中国农业科学院2019-07-05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：甘蓝论文; 雄性不育论文; 转录组论文; 蛋白组论文; 中国农业科学院2019-07-05论文;