本文主要研究内容
作者郭丹丹（2019）在《软枣猕猴桃（Actinidia arguta）性别相关分子标记的开发及应用》一文中研究指出：绝大多数猕猴桃属植物是功能性的雌雄异株植物,其实生个体童期长达5~7年。早期生产上栽培种植的品种主要来自于中华猕猴桃复合体,软枣猕猴桃是近年来迅速发展的新兴栽培种类。目前产业上雌株的经济价值和需求明显高于雄株,育种工作仍主要集中在雌性优良品种选育方面。因此,利用幼苗早期性别鉴定的技术或标记,在育种过程中进行实生苗早期性别鉴定,提前去除多余雄株,将节约大量的育种成本,加速育种进程。本研究首先利用PCR扩增技术,验证前人在中华猕猴桃或其相关群体中已开发的性别鉴定分子标记在本研究所用材料中的通用性;进一步利用全基因组重测序的方法,自行开发了软枣猕猴桃实生苗性别早期鉴定分子标记,以实现软枣猕猴桃植株性别的早期鉴定,并在中华猕猴桃中进行了通用性验证。主要研究结果如下:1.已报道猕猴桃性别分子标记的通用性验证。以已知性别的64份中华猕猴桃复合体材料和64份软枣猕猴桃材料,对前人已开发的两组猕猴桃性别鉴定分子标记,SCAR标记SmX和SmY1,以及SSR标记A001、A002和A003,进行通用性检验。结果表明:(1)在本研究所用的64份中华猕猴桃复合体样品中,SmX未表现出性别特异性,而SmY1能够很好地鉴定中华猕猴桃的性别,雄性植株性别鉴定准确率为96.55%、雌性准确率为100%,可广泛应用于中华猕猴桃原变种的性别鉴定;A001、A002和A003的性别鉴定准确率均较低,未表现出性别特异性。(2)对于本研究所用的64份软枣猕猴桃样品,SmX和SmY1均不能有效鉴定其性别,A001、A002和A003三个SSR标记也不具有性别特异性。(3)由于两组标记在软枣猕猴桃样品中均不能通用,因此有必要开发基于软枣猕猴桃实生苗性别早期鉴定的分子标记。2.软枣猕猴桃性别相关分子标记的开发及通用性验证。本研究以软枣猕猴桃人工杂交组合‘红宝石星’(♀)×11-17(雄株代号,经野生软枣猕猴桃植株种子实生选种所得)群体中已结果的实生后代雌雄各15株为材料,通过全基因组重测序以及BSA分析技术,以‘红阳’猕猴桃基因组为参考基因组进行生物信息学分析,开发性别鉴定分子标记。结果如下:(1)得到了性别差异序列g11.t2、g1.t1、g11.t3、g2.t2、g5.t1、g3.t1、g6.t1、g4.t1和g11.t1,分别对该9条序列设计引物,利用PCR扩增技术,筛选得到2对雄性特异性的引物Primer 11和Primer 51;在建库群体以及其他野生自然群体共128个已知性别的软枣猕猴桃样品中对Primer 11和Primer 51进行了PCR验证,两标记验证准确率分别为97.89%和97.03%。(2)分别对Primer 11和Primer 51的扩增产物进行测序,得到其产物序列L11和L51;在猕猴桃基因库(kiwifruit genome database)和Gene-bank中对L11和L51进行Blast分析后发现,该两条片段可能位于猕猴桃性染色体的雄性特异区域,但还需进行进一步定位。(3)在48份(16雄,32雌)中华猕猴桃复合体材料中验证两标记的通用性后发现,Primer 11能够得到相应的特异条带,雄性准确率为100%,但雌性验证结果准确率仅为68.8%,表明Primer 11在中华猕猴桃复合体性别鉴定中有一定的准确率,但仍需加大群体进一步验证;Primer 51在该种样品中未扩增出清晰的条带,表明其不适用于中华猕猴桃的性别鉴定。
Abstract
jue da duo shu mi hou tao shu zhi wu shi gong neng xing de ci xiong yi zhu zhi wu ,ji shi sheng ge ti tong ji chang da 5~7nian 。zao ji sheng chan shang zai pei chong zhi de pin chong zhu yao lai zi yu zhong hua mi hou tao fu ge ti ,ruan zao mi hou tao shi jin nian lai xun su fa zhan de xin xing zai pei chong lei 。mu qian chan ye shang ci zhu de jing ji jia zhi he xu qiu ming xian gao yu xiong zhu ,yo chong gong zuo reng zhu yao ji zhong zai ci xing you liang pin chong shua yo fang mian 。yin ci ,li yong you miao zao ji xing bie jian ding de ji shu huo biao ji ,zai yo chong guo cheng zhong jin hang shi sheng miao zao ji xing bie jian ding ,di qian qu chu duo yu xiong zhu ,jiang jie yao da liang de yo chong cheng ben ,jia su yo chong jin cheng 。ben yan jiu shou xian li yong PCRkuo zeng ji shu ,yan zheng qian ren zai zhong hua mi hou tao huo ji xiang guan qun ti zhong yi kai fa de xing bie jian ding fen zi biao ji zai ben yan jiu suo yong cai liao zhong de tong yong xing ;jin yi bu li yong quan ji yin zu chong ce xu de fang fa ,zi hang kai fa le ruan zao mi hou tao shi sheng miao xing bie zao ji jian ding fen zi biao ji ,yi shi xian ruan zao mi hou tao zhi zhu xing bie de zao ji jian ding ,bing zai zhong hua mi hou tao zhong jin hang le tong yong xing yan zheng 。zhu yao yan jiu jie guo ru xia :1.yi bao dao mi hou tao xing bie fen zi biao ji de tong yong xing yan zheng 。yi yi zhi xing bie de 64fen zhong hua mi hou tao fu ge ti cai liao he 64fen ruan zao mi hou tao cai liao ,dui qian ren yi kai fa de liang zu mi hou tao xing bie jian ding fen zi biao ji ,SCARbiao ji SmXhe SmY1,yi ji SSRbiao ji A001、A002he A003,jin hang tong yong xing jian yan 。jie guo biao ming :(1)zai ben yan jiu suo yong de 64fen zhong hua mi hou tao fu ge ti yang pin zhong ,SmXwei biao xian chu xing bie te yi xing ,er SmY1neng gou hen hao de jian ding zhong hua mi hou tao de xing bie ,xiong xing zhi zhu xing bie jian ding zhun que lv wei 96.55%、ci xing zhun que lv wei 100%,ke an fan ying yong yu zhong hua mi hou tao yuan bian chong de xing bie jian ding ;A001、A002he A003de xing bie jian ding zhun que lv jun jiao di ,wei biao xian chu xing bie te yi xing 。(2)dui yu ben yan jiu suo yong de 64fen ruan zao mi hou tao yang pin ,SmXhe SmY1jun bu neng you xiao jian ding ji xing bie ,A001、A002he A003san ge SSRbiao ji ye bu ju you xing bie te yi xing 。(3)you yu liang zu biao ji zai ruan zao mi hou tao yang pin zhong jun bu neng tong yong ,yin ci you bi yao kai fa ji yu ruan zao mi hou tao shi sheng miao xing bie zao ji jian ding de fen zi biao ji 。2.ruan zao mi hou tao xing bie xiang guan fen zi biao ji de kai fa ji tong yong xing yan zheng 。ben yan jiu yi ruan zao mi hou tao ren gong za jiao zu ge ‘gong bao dan xing ’(♀)×11-17(xiong zhu dai hao ,jing ye sheng ruan zao mi hou tao zhi zhu chong zi shi sheng shua chong suo de )qun ti zhong yi jie guo de shi sheng hou dai ci xiong ge 15zhu wei cai liao ,tong guo quan ji yin zu chong ce xu yi ji BSAfen xi ji shu ,yi ‘gong yang ’mi hou tao ji yin zu wei can kao ji yin zu jin hang sheng wu xin xi xue fen xi ,kai fa xing bie jian ding fen zi biao ji 。jie guo ru xia :(1)de dao le xing bie cha yi xu lie g11.t2、g1.t1、g11.t3、g2.t2、g5.t1、g3.t1、g6.t1、g4.t1he g11.t1,fen bie dui gai 9tiao xu lie she ji yin wu ,li yong PCRkuo zeng ji shu ,shai shua de dao 2dui xiong xing te yi xing de yin wu Primer 11he Primer 51;zai jian ku qun ti yi ji ji ta ye sheng zi ran qun ti gong 128ge yi zhi xing bie de ruan zao mi hou tao yang pin zhong dui Primer 11he Primer 51jin hang le PCRyan zheng ,liang biao ji yan zheng zhun que lv fen bie wei 97.89%he 97.03%。(2)fen bie dui Primer 11he Primer 51de kuo zeng chan wu jin hang ce xu ,de dao ji chan wu xu lie L11he L51;zai mi hou tao ji yin ku (kiwifruit genome database)he Gene-bankzhong dui L11he L51jin hang Blastfen xi hou fa xian ,gai liang tiao pian duan ke neng wei yu mi hou tao xing ran se ti de xiong xing te yi ou yu ,dan hai xu jin hang jin yi bu ding wei 。(3)zai 48fen (16xiong ,32ci )zhong hua mi hou tao fu ge ti cai liao zhong yan zheng liang biao ji de tong yong xing hou fa xian ,Primer 11neng gou de dao xiang ying de te yi tiao dai ,xiong xing zhun que lv wei 100%,dan ci xing yan zheng jie guo zhun que lv jin wei 68.8%,biao ming Primer 11zai zhong hua mi hou tao fu ge ti xing bie jian ding zhong you yi ding de zhun que lv ,dan reng xu jia da qun ti jin yi bu yan zheng ;Primer 51zai gai chong yang pin zhong wei kuo zeng chu qing xi de tiao dai ,biao ming ji bu kuo yong yu zhong hua mi hou tao de xing bie jian ding 。
论文参考文献
论文详细介绍
论文作者分别是来自中国农业科学院的郭丹丹，发表于刊物中国农业科学院2019-07-05论文，是一篇关于软枣猕猴桃论文,中华猕猴桃复合体论文,分子标记论文,性别鉴定论文,通用性验证论文,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：软枣猕猴桃论文; 中华猕猴桃复合体论文; 分子标记论文; 性别鉴定论文; 通用性验证论文; 中国农业科学院2019-07-05论文;