本文主要研究内容
作者徐悦（2019）在《菠菜（Spinacia oleracea L.）毛状根诱导与高效基因组编辑体系的建立》一文中研究指出：发根农杆菌(Agrobacterium rhizgones)侵染植物后,可诱导植物产生毛状根。菠菜(Spinacia oleracea L.)是常见的食用蔬菜,目前尚未有菠菜毛状根的研究报道。本研究筛选到适合诱导菠菜毛状根的发根农杆菌菌株LBA9402。结果表明,LBA9402侵染菠菜外植体茎后,毛状根的诱导率最高可达16%。菠菜毛状根呈白色,具有丰富的根毛,能在无外源激素的固体培养基上快速增殖生长。通过诱导菠菜毛状根产生愈伤并进行分化,获得了菠菜毛状根的再生株。此外,本研究实现了发根农杆菌的Ri质粒与携带外源GFP基因的Ti质粒的共转化,PCR结果和荧光显微镜观察显示,发根农杆菌Ri质粒的rol B基因以及Ti质粒中GFP基因已在菠菜毛状根基因组中稳定表达。CRISPR/Cas9是一种基因组高效定点突变的工具,已广泛应用于模式植物和各种作物。尽管菠菜的基因组序列被公布,但CRISPR/Cas9技术尚未成功地应用于菠菜。在上面研究的基础上,我们以菠菜的毛状根为研究对象,使用CRISPR/Cas9精确编辑两个类纤维素合成酶 D 基因(Cellulose Synthase-Like D genes CSLD),Spo2331和Spo10340,探究其参与菠菜毛状根根毛形成的机制。结果表明,CRISPR/as9可在菠菜中形成四种突变类型,即置换,插入,删除和组合突变,其中删除突变所占比重最高,约占64.1%。每个靶位点之间的突变率具有很大差异。15个Sp02331独立的转基因毛状根系中,有7个是纯合或双等位基因突变根系,其他8个是杂合突变根系,所有纯合突变根系显示出根毛膨胀和变短的表型,与拟南芥csld2突变体的表型特征相似。获得的15个Spo10340独立的转基因发根系中,有13个杂合突变系根系,但没有获得纯合或双等位基因突变根系,所有杂合突变根系与正常毛状根无明显差别。菠菜CRISPR/Cas9系统的建立为菠菜的功能基因的鉴定以及菠菜的分子育种奠定了基础,在菠菜的基础研究和应用研究领域均具有重要意义。
Abstract
fa gen nong gan jun (Agrobacterium rhizgones)qin ran zhi wu hou ,ke you dao zhi wu chan sheng mao zhuang gen 。bo cai (Spinacia oleracea L.)shi chang jian de shi yong shu cai ,mu qian shang wei you bo cai mao zhuang gen de yan jiu bao dao 。ben yan jiu shai shua dao kuo ge you dao bo cai mao zhuang gen de fa gen nong gan jun jun zhu LBA9402。jie guo biao ming ,LBA9402qin ran bo cai wai zhi ti jing hou ,mao zhuang gen de you dao lv zui gao ke da 16%。bo cai mao zhuang gen cheng bai se ,ju you feng fu de gen mao ,neng zai mo wai yuan ji su de gu ti pei yang ji shang kuai su zeng shi sheng chang 。tong guo you dao bo cai mao zhuang gen chan sheng yu shang bing jin hang fen hua ,huo de le bo cai mao zhuang gen de zai sheng zhu 。ci wai ,ben yan jiu shi xian le fa gen nong gan jun de Rizhi li yu xie dai wai yuan GFPji yin de Tizhi li de gong zhuai hua ,PCRjie guo he ying guang xian wei jing guan cha xian shi ,fa gen nong gan jun Rizhi li de rol Bji yin yi ji Tizhi li zhong GFPji yin yi zai bo cai mao zhuang gen ji yin zu zhong wen ding biao da 。CRISPR/Cas9shi yi chong ji yin zu gao xiao ding dian tu bian de gong ju ,yi an fan ying yong yu mo shi zhi wu he ge chong zuo wu 。jin guan bo cai de ji yin zu xu lie bei gong bu ,dan CRISPR/Cas9ji shu shang wei cheng gong de ying yong yu bo cai 。zai shang mian yan jiu de ji chu shang ,wo men yi bo cai de mao zhuang gen wei yan jiu dui xiang ,shi yong CRISPR/Cas9jing que bian ji liang ge lei qian wei su ge cheng mei D ji yin (Cellulose Synthase-Like D genes CSLD),Spo2331he Spo10340,tan jiu ji can yu bo cai mao zhuang gen gen mao xing cheng de ji zhi 。jie guo biao ming ,CRISPR/as9ke zai bo cai zhong xing cheng si chong tu bian lei xing ,ji zhi huan ,cha ru ,shan chu he zu ge tu bian ,ji zhong shan chu tu bian suo zhan bi chong zui gao ,yao zhan 64.1%。mei ge ba wei dian zhi jian de tu bian lv ju you hen da cha yi 。15ge Sp02331du li de zhuai ji yin mao zhuang gen ji zhong ,you 7ge shi chun ge huo shuang deng wei ji yin tu bian gen ji ,ji ta 8ge shi za ge tu bian gen ji ,suo you chun ge tu bian gen ji xian shi chu gen mao peng zhang he bian duan de biao xing ,yu ni na gai csld2tu bian ti de biao xing te zheng xiang shi 。huo de de 15ge Spo10340du li de zhuai ji yin fa gen ji zhong ,you 13ge za ge tu bian ji gen ji ,dan mei you huo de chun ge huo shuang deng wei ji yin tu bian gen ji ,suo you za ge tu bian gen ji yu zheng chang mao zhuang gen mo ming xian cha bie 。bo cai CRISPR/Cas9ji tong de jian li wei bo cai de gong neng ji yin de jian ding yi ji bo cai de fen zi yo chong dian ding le ji chu ,zai bo cai de ji chu yan jiu he ying yong yan jiu ling yu jun ju you chong yao yi yi 。
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论文作者分别是来自东北林业大学的徐悦，发表于刊物东北林业大学2019-11-18论文，是一篇关于菠菜论文,毛状根论文,植物再生体系论文,根毛论文,东北林业大学2019-11-18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，资料来自东北林业大学2019-11-18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：菠菜论文; 毛状根论文; 植物再生体系论文; 根毛论文; 东北林业大学2019-11-18论文;