本文主要研究内容
作者李俊香（2019）在《甜瓜棒孢叶斑病菌鉴定及农杆菌介导的遗传转化体系建立》一文中研究指出：甜瓜（Cucumis melo L.）是我国最重要的葫芦科作物之一,栽培广泛,具有重要的经济价值。为害甜瓜的病害多达数十种,其中真菌病害最为普遍,侵染的真菌种类也较多,而且仍然存在新突发病害。本文鉴定一种甜瓜叶斑病,研究该病菌的生物学特性,筛选针对该病菌的杀菌剂,以期指导该病害的防控;同时应用农杆菌介导方法,创建该病菌的突变体库,为致病基因研究奠定基础。取得的主要研究结果如下:1、甜瓜棒孢叶斑病菌鉴定2015年10月在我国广西武鸣甜瓜主要种植区进行病害调查时发现一种叶斑病。将病样分离病原真菌,通过形态学观察、多基因鉴定和致病性试验鉴定病原菌。在PDA平板上,病原菌菌落圆形,中间墨绿色,边缘浅褐色,气生菌丝茂盛。分生孢子呈圆柱形或倒棍棒形,单生或串生,直立或稍微弯曲,具0-13个隔膜,大小为3.1-6.4×14-138.9μm。通过克隆并测定该菌的4个保守序列ITS1,2、ITS4,5、ACT和TUB2,说明该菌为多主棒孢。甜瓜叶片上分离纯化的病原菌回接到甜瓜幼苗,表现出与病害调查时田间相似的症状并再次分离到同一病原菌。该菌通常危害植物叶片引起叶斑病,寄主广泛。接种该病原菌的西瓜（Citrullus lanatus）和黄瓜（Cucumis sativus L.）幼苗及离体叶片也可以发病。病原菌菌丝生长的适宜碳源是可溶性淀粉和葡萄糖,适宜氮源是KNO3、NaNO3和蛋白胨,适宜温度是25-29°C,适宜pH是6-9。适宜分生孢子萌发的碳源是蔗糖和葡萄糖,pH是5-8。2、甜瓜棒孢叶斑病菌杀菌剂筛选为筛选甜瓜棒孢叶斑病菌的高效杀菌剂,在室内采用菌丝生长速率法测定22种杀菌剂对甜瓜棒孢叶斑病菌Cc-SY和Cc-12菌株菌丝生长的抑制能力。结果表明,50%咪鲜胺锰盐WP对甜瓜棒孢叶斑病菌Cc-SY和Cc-12菌株的EC50都最小,病菌对它的敏感性最高,抑菌效果最好;30%苯醚甲环唑SC、40%氟硅唑EC和80%福美双WG对甜瓜棒孢叶斑病菌Cc-12菌株的EC50均小于5μg/mL,抑菌效果较好,但在Cc-SY菌株上抑菌效果减弱。在试验浓度下,2个甜瓜棒孢叶斑病菌供试菌株对25%嘧菌酯SC、75%百菌清WP和90%多菌灵WG都非常不敏感,说明这3种杀菌剂对甜瓜棒孢叶斑病的防控不能起到有效作用。3、甜瓜棒孢叶斑病菌全基因组测序对多主棒孢甜瓜分离物Cc-12菌株进行了全基因组测序,获得了大小为44 M的基因组序列。以模式真菌的分化时间作为校正时间,估算C.cassiicola的分化时间为82.5 Mya。细胞降解酶是真菌成功侵染并定殖寄主的重要因子,注释结果显示葡萄糖苷水解酶有351个。4、农杆菌介导的多主棒孢遗传转化本研究利用ATMT转化技术,将T-DNA随机插入到甜瓜棒孢叶斑病菌基因组中,获得了具有潮霉素抗性的甜瓜棒孢叶斑病菌T-DNA插入突变体,初步建立了稳定高效的根癌农杆菌介导的甜瓜棒孢叶斑病菌遗传转化体系。同时,优化转化效率的影响因素,转化效率可以达到每106个分生孢子转化获得102-148个转化子。这为获得致病突变体材料提供潜在可能,为深入研究多主棒孢菌的致病机制奠定基础。突变体分子鉴定检测到潮霉素磷酸转移酶基因片段特异条带,Southern blot杂交结果表明T-DNA是随机整合到Cc-12菌株基因组中的,突变体单拷贝比例为90%。表型变异突变体对其进行致病性试验。筛选出3株无致病力突变体（CT0001,CT0022和CT0013）,1株致病力严重减弱突变体（CT0005）。利用TAIL-PCR技术和多主棒孢的基因组数据,进一步获得了这4株单拷贝插入低致病力突变体T-DNA的侧翼序列。这些结果为进一步鉴定致病相关基因奠定了基础。
Abstract
tian gua （Cucumis melo L.）shi wo guo zui chong yao de hu lu ke zuo wu zhi yi ,zai pei an fan ,ju you chong yao de jing ji jia zhi 。wei hai tian gua de bing hai duo da shu shi chong ,ji zhong zhen jun bing hai zui wei pu bian ,qin ran de zhen jun chong lei ye jiao duo ,er ju reng ran cun zai xin tu fa bing hai 。ben wen jian ding yi chong tian gua xie ban bing ,yan jiu gai bing jun de sheng wu xue te xing ,shai shua zhen dui gai bing jun de sha jun ji ,yi ji zhi dao gai bing hai de fang kong ;tong shi ying yong nong gan jun jie dao fang fa ,chuang jian gai bing jun de tu bian ti ku ,wei zhi bing ji yin yan jiu dian ding ji chu 。qu de de zhu yao yan jiu jie guo ru xia :1、tian gua bang bao xie ban bing jun jian ding 2015nian 10yue zai wo guo an xi wu ming tian gua zhu yao chong zhi ou jin hang bing hai diao cha shi fa xian yi chong xie ban bing 。jiang bing yang fen li bing yuan zhen jun ,tong guo xing tai xue guan cha 、duo ji yin jian ding he zhi bing xing shi yan jian ding bing yuan jun 。zai PDAping ban shang ,bing yuan jun jun la yuan xing ,zhong jian mo lu se ,bian yuan jian he se ,qi sheng jun si mao cheng 。fen sheng bao zi cheng yuan zhu xing huo dao gun bang xing ,chan sheng huo chuan sheng ,zhi li huo shao wei wan qu ,ju 0-13ge ge mo ,da xiao wei 3.1-6.4×14-138.9μm。tong guo ke long bing ce ding gai jun de 4ge bao shou xu lie ITS1,2、ITS4,5、ACThe TUB2,shui ming gai jun wei duo zhu bang bao 。tian gua xie pian shang fen li chun hua de bing yuan jun hui jie dao tian gua you miao ,biao xian chu yu bing hai diao cha shi tian jian xiang shi de zheng zhuang bing zai ci fen li dao tong yi bing yuan jun 。gai jun tong chang wei hai zhi wu xie pian yin qi xie ban bing ,ji zhu an fan 。jie chong gai bing yuan jun de xi gua （Citrullus lanatus）he huang gua （Cucumis sativus L.）you miao ji li ti xie pian ye ke yi fa bing 。bing yuan jun jun si sheng chang de kuo yi tan yuan shi ke rong xing dian fen he pu tao tang ,kuo yi dan yuan shi KNO3、NaNO3he dan bai dong ,kuo yi wen du shi 25-29°C,kuo yi pHshi 6-9。kuo yi fen sheng bao zi meng fa de tan yuan shi zhe tang he pu tao tang ,pHshi 5-8。2、tian gua bang bao xie ban bing jun sha jun ji shai shua wei shai shua tian gua bang bao xie ban bing jun de gao xiao sha jun ji ,zai shi nei cai yong jun si sheng chang su lv fa ce ding 22chong sha jun ji dui tian gua bang bao xie ban bing jun Cc-SYhe Cc-12jun zhu jun si sheng chang de yi zhi neng li 。jie guo biao ming ,50%mi xian an meng yan WPdui tian gua bang bao xie ban bing jun Cc-SYhe Cc-12jun zhu de EC50dou zui xiao ,bing jun dui ta de min gan xing zui gao ,yi jun xiao guo zui hao ;30%ben mi jia huan zuo SC、40%fu gui zuo EChe 80%fu mei shuang WGdui tian gua bang bao xie ban bing jun Cc-12jun zhu de EC50jun xiao yu 5μg/mL,yi jun xiao guo jiao hao ,dan zai Cc-SYjun zhu shang yi jun xiao guo jian ruo 。zai shi yan nong du xia ,2ge tian gua bang bao xie ban bing jun gong shi jun zhu dui 25%mi jun zhi SC、75%bai jun qing WPhe 90%duo jun ling WGdou fei chang bu min gan ,shui ming zhe 3chong sha jun ji dui tian gua bang bao xie ban bing de fang kong bu neng qi dao you xiao zuo yong 。3、tian gua bang bao xie ban bing jun quan ji yin zu ce xu dui duo zhu bang bao tian gua fen li wu Cc-12jun zhu jin hang le quan ji yin zu ce xu ,huo de le da xiao wei 44 Mde ji yin zu xu lie 。yi mo shi zhen jun de fen hua shi jian zuo wei jiao zheng shi jian ,gu suan C.cassiicolade fen hua shi jian wei 82.5 Mya。xi bao jiang jie mei shi zhen jun cheng gong qin ran bing ding shi ji zhu de chong yao yin zi ,zhu shi jie guo xian shi pu tao tang gan shui jie mei you 351ge 。4、nong gan jun jie dao de duo zhu bang bao wei chuan zhuai hua ben yan jiu li yong ATMTzhuai hua ji shu ,jiang T-DNAsui ji cha ru dao tian gua bang bao xie ban bing jun ji yin zu zhong ,huo de le ju you chao mei su kang xing de tian gua bang bao xie ban bing jun T-DNAcha ru tu bian ti ,chu bu jian li le wen ding gao xiao de gen ai nong gan jun jie dao de tian gua bang bao xie ban bing jun wei chuan zhuai hua ti ji 。tong shi ,you hua zhuai hua xiao lv de ying xiang yin su ,zhuai hua xiao lv ke yi da dao mei 106ge fen sheng bao zi zhuai hua huo de 102-148ge zhuai hua zi 。zhe wei huo de zhi bing tu bian ti cai liao di gong qian zai ke neng ,wei shen ru yan jiu duo zhu bang bao jun de zhi bing ji zhi dian ding ji chu 。tu bian ti fen zi jian ding jian ce dao chao mei su lin suan zhuai yi mei ji yin pian duan te yi tiao dai ,Southern blotza jiao jie guo biao ming T-DNAshi sui ji zheng ge dao Cc-12jun zhu ji yin zu zhong de ,tu bian ti chan kao bei bi li wei 90%。biao xing bian yi tu bian ti dui ji jin hang zhi bing xing shi yan 。shai shua chu 3zhu mo zhi bing li tu bian ti （CT0001,CT0022he CT0013）,1zhu zhi bing li yan chong jian ruo tu bian ti （CT0005）。li yong TAIL-PCRji shu he duo zhu bang bao de ji yin zu shu ju ,jin yi bu huo de le zhe 4zhu chan kao bei cha ru di zhi bing li tu bian ti T-DNAde ce yi xu lie 。zhe xie jie guo wei jin yi bu jian ding zhi bing xiang guan ji yin dian ding le ji chu 。
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论文详细介绍
论文作者分别是来自华中农业大学的李俊香，发表于刊物华中农业大学2019-07-15论文，是一篇关于多主棒孢论文,甜瓜论文,生物学特性论文,杀菌剂论文,基因组测序论文,根癌农杆菌介导的转化论文,华中农业大学2019-07-15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，资料来自华中农业大学2019-07-15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：多主棒孢论文; 甜瓜论文; 生物学特性论文; 杀菌剂论文; 基因组测序论文; 根癌农杆菌介导的转化论文; 华中农业大学2019-07-15论文;