本文主要研究内容
作者岳松青（2019）在《苹果砧木对镉的吸收和忍耐及相关基因MhNRAMP1克隆和功能鉴定》一文中研究指出：果树是多年生植物,即使在重金属含量较低的土壤中,其根系也会吸收和积累较多重金属;镉是环境中毒性最强,也是目前对我国耕地污染最严重的重金属之一。苹果是全球四大水果之一,其根系来自砧木。本研究以六种苹果砧木（小金海棠、新疆野苹果、崂山柰子、八棱海棠、山定子和平邑甜茶）为试材,分析比较它们对镉的吸收、转运及耐受性,并从平邑甜茶分离克隆MhNRAMP1基因,探讨转MhNRAMP1酵母和苹果愈伤组织对镉的吸收和忍耐情况,结果表明:1.六种苹果砧木Cd含量存在显著差异,根系Cd含量远远高于地上部分。山定子地上部和根部Cd含量都最低,Cd2+吸收速率、转运系数（Tf）最小,山定子中与镉吸收相关基因MdZIPT1、MdIRT1和MdNRAMP1的表达量也低于其他砧木;新疆野苹果中MdZIPT1、MdIRT1和MdNRAMP1的表达量最高,Cd2+吸收速率和积累量也最高。除了新疆野苹果,其他5种砧木与对照相比向地上部转运Cd2+相关基因MdHMA3的表达量都表现为下调。由此发现Cd胁迫下苹果砧木幼苗对Cd2+吸收、积累情况与基因的表达相一致。以上结果表明,50μM CdSO4处理下山定子对Cd的吸收和转运能力小于其他5种苹果砧木。2.50μM CdSO4处理14 d后,新疆野苹果的根长、鲜重和根系活力下降幅度、细胞死亡率增加幅度、MDA含量和叶片H2O2含量的增加幅度均最高,山定子干重下降幅度和叶片H2O2含量增加幅度最低,八棱海棠根系细胞死亡率增加幅度最低;八棱海棠根系POD和SOD活性提高幅度最大,崂山柰子叶绿素相对含量降低幅度最大,八棱海棠和山定子变化不显著。在主成分分析的基础上,利用隶属函数对各砧木所测指标进行综合分析显示,六种苹果砧木由强到弱的耐镉能力顺序为:八棱海棠>山定子>平邑甜茶>崂山柰子>小金海棠>新疆野苹果。3.MhNRAMP1基因,它的开放阅读框（ORF）长度为1638 bp,编码含有545个氨基酸的蛋白,预测其蛋白质分子量为59.12kD,定位于质膜,并具有12个跨膜结构;利用瞬时转化烟草叶片技术,基因MhNRAMP1被鉴定出特异定位于质膜处,是一种膜蛋白;在根、茎、叶中都能表达且具有特异性,其中根部表达最高,其次是叶,茎部最低。在根部的表达量随镉胁迫明显上调,在茎中略有上调,而在叶片部位其表达量下降较大。4.在镉处理下表达MhNRAMP1基因的酵母在点板试验和生长曲线测试中表现为长势较差,受到更为严重的毒害作用,验证了MhNRAMP1基因的表达会使酵母的镉敏感性增加。进一步测定酵母的镉含量,结果也显示与空载酵母相比,MhNRAMP1基因能够显著提高酵母中镉的积累量。综上充分说明MhNRAMP1基因在酵母中参与了镉的吸收,促进了镉的积累,提高了酵母的镉敏感性。5.在镉处理情况下,超表达基因MhNRAMP1的苹果愈伤,鲜重下降,电导率上升;利用镉荧光染色技术和MS–ICP检测苹果愈伤中的镉含量,结果均显示超表达苹果愈伤镉含量更高;利用非损伤微测技术分别检测了野生型和转基因型苹果愈伤的镉吸收速率,超表达苹果愈伤对镉的吸收速率均强于野生型。超表达基因MhNRAMP1提高了苹果愈伤的镉敏感性和镉含量,降低了其耐镉性。
Abstract
guo shu shi duo nian sheng zhi wu ,ji shi zai chong jin shu han liang jiao di de tu rang zhong ,ji gen ji ye hui xi shou he ji lei jiao duo chong jin shu ;ge shi huan jing zhong du xing zui jiang ,ye shi mu qian dui wo guo geng de wu ran zui yan chong de chong jin shu zhi yi 。ping guo shi quan qiu si da shui guo zhi yi ,ji gen ji lai zi zhen mu 。ben yan jiu yi liu chong ping guo zhen mu （xiao jin hai tang 、xin jiang ye ping guo 、lao shan nai zi 、ba leng hai tang 、shan ding zi he ping yi tian cha ）wei shi cai ,fen xi bi jiao ta men dui ge de xi shou 、zhuai yun ji nai shou xing ,bing cong ping yi tian cha fen li ke long MhNRAMP1ji yin ,tan tao zhuai MhNRAMP1jiao mu he ping guo yu shang zu zhi dui ge de xi shou he ren nai qing kuang ,jie guo biao ming :1.liu chong ping guo zhen mu Cdhan liang cun zai xian zhe cha yi ,gen ji Cdhan liang yuan yuan gao yu de shang bu fen 。shan ding zi de shang bu he gen bu Cdhan liang dou zui di ,Cd2+xi shou su lv 、zhuai yun ji shu （Tf）zui xiao ,shan ding zi zhong yu ge xi shou xiang guan ji yin MdZIPT1、MdIRT1he MdNRAMP1de biao da liang ye di yu ji ta zhen mu ;xin jiang ye ping guo zhong MdZIPT1、MdIRT1he MdNRAMP1de biao da liang zui gao ,Cd2+xi shou su lv he ji lei liang ye zui gao 。chu le xin jiang ye ping guo ,ji ta 5chong zhen mu yu dui zhao xiang bi xiang de shang bu zhuai yun Cd2+xiang guan ji yin MdHMA3de biao da liang dou biao xian wei xia diao 。you ci fa xian Cdxie pai xia ping guo zhen mu you miao dui Cd2+xi shou 、ji lei qing kuang yu ji yin de biao da xiang yi zhi 。yi shang jie guo biao ming ,50μM CdSO4chu li xia shan ding zi dui Cdde xi shou he zhuai yun neng li xiao yu ji ta 5chong ping guo zhen mu 。2.50μM CdSO4chu li 14 dhou ,xin jiang ye ping guo de gen chang 、xian chong he gen ji huo li xia jiang fu du 、xi bao si wang lv zeng jia fu du 、MDAhan liang he xie pian H2O2han liang de zeng jia fu du jun zui gao ,shan ding zi gan chong xia jiang fu du he xie pian H2O2han liang zeng jia fu du zui di ,ba leng hai tang gen ji xi bao si wang lv zeng jia fu du zui di ;ba leng hai tang gen ji PODhe SODhuo xing di gao fu du zui da ,lao shan nai zi xie lu su xiang dui han liang jiang di fu du zui da ,ba leng hai tang he shan ding zi bian hua bu xian zhe 。zai zhu cheng fen fen xi de ji chu shang ,li yong li shu han shu dui ge zhen mu suo ce zhi biao jin hang zeng ge fen xi xian shi ,liu chong ping guo zhen mu you jiang dao ruo de nai ge neng li shun xu wei :ba leng hai tang >shan ding zi >ping yi tian cha >lao shan nai zi >xiao jin hai tang >xin jiang ye ping guo 。3.MhNRAMP1ji yin ,ta de kai fang yue dou kuang （ORF）chang du wei 1638 bp,bian ma han you 545ge an ji suan de dan bai ,yu ce ji dan bai zhi fen zi liang wei 59.12kD,ding wei yu zhi mo ,bing ju you 12ge kua mo jie gou ;li yong shun shi zhuai hua yan cao xie pian ji shu ,ji yin MhNRAMP1bei jian ding chu te yi ding wei yu zhi mo chu ,shi yi chong mo dan bai ;zai gen 、jing 、xie zhong dou neng biao da ju ju you te yi xing ,ji zhong gen bu biao da zui gao ,ji ci shi xie ,jing bu zui di 。zai gen bu de biao da liang sui ge xie pai ming xian shang diao ,zai jing zhong lve you shang diao ,er zai xie pian bu wei ji biao da liang xia jiang jiao da 。4.zai ge chu li xia biao da MhNRAMP1ji yin de jiao mu zai dian ban shi yan he sheng chang qu xian ce shi zhong biao xian wei chang shi jiao cha ,shou dao geng wei yan chong de du hai zuo yong ,yan zheng le MhNRAMP1ji yin de biao da hui shi jiao mu de ge min gan xing zeng jia 。jin yi bu ce ding jiao mu de ge han liang ,jie guo ye xian shi yu kong zai jiao mu xiang bi ,MhNRAMP1ji yin neng gou xian zhe di gao jiao mu zhong ge de ji lei liang 。zeng shang chong fen shui ming MhNRAMP1ji yin zai jiao mu zhong can yu le ge de xi shou ,cu jin le ge de ji lei ,di gao le jiao mu de ge min gan xing 。5.zai ge chu li qing kuang xia ,chao biao da ji yin MhNRAMP1de ping guo yu shang ,xian chong xia jiang ,dian dao lv shang sheng ;li yong ge ying guang ran se ji shu he MS–ICPjian ce ping guo yu shang zhong de ge han liang ,jie guo jun xian shi chao biao da ping guo yu shang ge han liang geng gao ;li yong fei sun shang wei ce ji shu fen bie jian ce le ye sheng xing he zhuai ji yin xing ping guo yu shang de ge xi shou su lv ,chao biao da ping guo yu shang dui ge de xi shou su lv jun jiang yu ye sheng xing 。chao biao da ji yin MhNRAMP1di gao le ping guo yu shang de ge min gan xing he ge han liang ,jiang di le ji nai ge xing 。
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论文作者分别是来自山东农业大学的岳松青，发表于刊物山东农业大学2019-07-08论文，是一篇关于苹果砧木论文,镉吸收论文,基因克隆论文,功能鉴定论文,山东农业大学2019-07-08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，资料来自山东农业大学2019-07-08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：苹果砧木论文; 镉吸收论文; 基因克隆论文; 功能鉴定论文; 山东农业大学2019-07-08论文;