本文主要研究内容
作者陈珊珊（2019）在《番茄表皮毛形态和内部细胞骨架排布的研究》一文中研究指出：番茄(Solanum lycopersicum Mill.)是世界范围内广泛栽培的蔬菜作物,也是一种重要的经济作物。覆盖于番茄植株表面的表皮毛(trichome)作为番茄应对环境胁迫的第一道屏障,可通过空间障碍、物理诱捕和化学毒杀等方式阻碍病毒或病原菌的传播,减少和抵御昆虫对植物体的危害,增强植株对非生物逆境的抗性,对于番茄的广谱抗性具有重要作用。随着细胞生物学的发展,细胞骨架(cytoskeleton)在拟南芥表皮毛发育过程中的动态变化得到了充分的研究。KCBP(kinesin-like calmodulin-binding protein)作为中心枢纽蛋白同时结合微管(microtubules,MTs)与微丝(actin filaments,AFs),从而调控拟南芥表皮毛形态建成的关键功能已在模式植物拟南芥中揭示,但相关研究在番茄中仍鲜有报道。因此,通过细胞生物学手段研究番茄表皮毛内部细胞骨架的排布,鉴定番茄表皮毛发育过程中的关键基因,解析番茄表皮毛发育的分子机制,对于提高番茄植株的广谱抗性具有重要的理论意义和实际应用价值。本研究构建了增强绿色荧光蛋白(enhanced green fluorescent protein,EGFP)标记微管或微丝细胞骨架的转基因材料,以及敲除SlKCBP的基因编辑材料,利用激光共聚焦显微镜观察了番茄表皮毛中细胞骨架的排布状态,并进一步研究了 SlKCBP在番茄表皮毛发育过程中的作用。主要研究结果如下:1.构建EGFP标记微管或微丝的转基因番茄材料SlTub6-EGFP和Lifeact-EGFP,并利用该材料探索利用激光共聚焦显微镜观察番茄细胞骨架的方法,成功观察到番茄表皮细胞和表皮毛内部的细胞骨架排布,为在番茄中开展细胞骨架相关研究创造了基础材料。2.观察SlTub6-EGFP、Lifeact-EGFP转基因材料的表皮毛类型和形态,结果表明标记了微管或微丝的转基因株系的表皮毛类型和形态与野生型无差异,为细胞骨架标记材料的广泛利用提供可行性依据。利用激光共聚焦显微镜观察番茄表皮毛内部的细胞骨架排布,观察到微管在表皮毛的横断面上呈环状排布,而微丝则表现为径向排布,呈束状。3.通过生物信息学方法鉴定、分析番茄中的KCBP蛋白,并进一步利用CRISPR/Cas9系统对番茄基因组进行精确编辑,敲除SlKCBP基因,获得SlKCBP-CRISPR材料。SlKCBP-CRISPR材料在表皮毛的类型及形态上与野生型无明显差异。通过杂交手段,将SlKCBP-CRISPR材料分别与SlTub6-EGFP、Lifeact-EGFP杂交,获得EGFP标记微管或微丝的SlKCBP基因编辑材料,为深入研究SlKCBP基因在细胞骨架排布中的作用奠定基础。
Abstract
fan jia (Solanum lycopersicum Mill.)shi shi jie fan wei nei an fan zai pei de shu cai zuo wu ,ye shi yi chong chong yao de jing ji zuo wu 。fu gai yu fan jia zhi zhu biao mian de biao pi mao (trichome)zuo wei fan jia ying dui huan jing xie pai de di yi dao bing zhang ,ke tong guo kong jian zhang ai 、wu li you bu he hua xue du sha deng fang shi zu ai bing du huo bing yuan jun de chuan bo ,jian shao he di yu kun chong dui zhi wu ti de wei hai ,zeng jiang zhi zhu dui fei sheng wu ni jing de kang xing ,dui yu fan jia de an pu kang xing ju you chong yao zuo yong 。sui zhao xi bao sheng wu xue de fa zhan ,xi bao gu jia (cytoskeleton)zai ni na gai biao pi mao fa yo guo cheng zhong de dong tai bian hua de dao le chong fen de yan jiu 。KCBP(kinesin-like calmodulin-binding protein)zuo wei zhong xin shu niu dan bai tong shi jie ge wei guan (microtubules,MTs)yu wei si (actin filaments,AFs),cong er diao kong ni na gai biao pi mao xing tai jian cheng de guan jian gong neng yi zai mo shi zhi wu ni na gai zhong jie shi ,dan xiang guan yan jiu zai fan jia zhong reng xian you bao dao 。yin ci ,tong guo xi bao sheng wu xue shou duan yan jiu fan jia biao pi mao nei bu xi bao gu jia de pai bu ,jian ding fan jia biao pi mao fa yo guo cheng zhong de guan jian ji yin ,jie xi fan jia biao pi mao fa yo de fen zi ji zhi ,dui yu di gao fan jia zhi zhu de an pu kang xing ju you chong yao de li lun yi yi he shi ji ying yong jia zhi 。ben yan jiu gou jian le zeng jiang lu se ying guang dan bai (enhanced green fluorescent protein,EGFP)biao ji wei guan huo wei si xi bao gu jia de zhuai ji yin cai liao ,yi ji qiao chu SlKCBPde ji yin bian ji cai liao ,li yong ji guang gong ju jiao xian wei jing guan cha le fan jia biao pi mao zhong xi bao gu jia de pai bu zhuang tai ,bing jin yi bu yan jiu le SlKCBPzai fan jia biao pi mao fa yo guo cheng zhong de zuo yong 。zhu yao yan jiu jie guo ru xia :1.gou jian EGFPbiao ji wei guan huo wei si de zhuai ji yin fan jia cai liao SlTub6-EGFPhe Lifeact-EGFP,bing li yong gai cai liao tan suo li yong ji guang gong ju jiao xian wei jing guan cha fan jia xi bao gu jia de fang fa ,cheng gong guan cha dao fan jia biao pi xi bao he biao pi mao nei bu de xi bao gu jia pai bu ,wei zai fan jia zhong kai zhan xi bao gu jia xiang guan yan jiu chuang zao le ji chu cai liao 。2.guan cha SlTub6-EGFP、Lifeact-EGFPzhuai ji yin cai liao de biao pi mao lei xing he xing tai ,jie guo biao ming biao ji le wei guan huo wei si de zhuai ji yin zhu ji de biao pi mao lei xing he xing tai yu ye sheng xing mo cha yi ,wei xi bao gu jia biao ji cai liao de an fan li yong di gong ke hang xing yi ju 。li yong ji guang gong ju jiao xian wei jing guan cha fan jia biao pi mao nei bu de xi bao gu jia pai bu ,guan cha dao wei guan zai biao pi mao de heng duan mian shang cheng huan zhuang pai bu ,er wei si ze biao xian wei jing xiang pai bu ,cheng shu zhuang 。3.tong guo sheng wu xin xi xue fang fa jian ding 、fen xi fan jia zhong de KCBPdan bai ,bing jin yi bu li yong CRISPR/Cas9ji tong dui fan jia ji yin zu jin hang jing que bian ji ,qiao chu SlKCBPji yin ,huo de SlKCBP-CRISPRcai liao 。SlKCBP-CRISPRcai liao zai biao pi mao de lei xing ji xing tai shang yu ye sheng xing mo ming xian cha yi 。tong guo za jiao shou duan ,jiang SlKCBP-CRISPRcai liao fen bie yu SlTub6-EGFP、Lifeact-EGFPza jiao ,huo de EGFPbiao ji wei guan huo wei si de SlKCBPji yin bian ji cai liao ,wei shen ru yan jiu SlKCBPji yin zai xi bao gu jia pai bu zhong de zuo yong dian ding ji chu 。
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论文详细介绍
论文作者分别是来自浙江大学的陈珊珊，发表于刊物浙江大学2019-05-20论文，是一篇关于番茄论文,表皮毛论文,细胞骨架论文,微管论文,微丝论文,浙江大学2019-05-20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，资料来自浙江大学2019-05-20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。