转NtFer1基因粳稻空育131抗Fe^(2+)胁迫能力分析

Analysis on Fe^(2+) Stress Resistance of Transgenic Japonica Kongyu 131 with NtFer1

旨在研究烟草铁蛋白基因NtFer1功能,提高粳稻抗逆性等。利用农杆菌介导法将NtFer1基因转入粳稻空育131,经抗性筛选、PCR、Southern杂交、Northern杂交和RT-PCR检测等,表明外源基因已整合到空育131基因组中,并能够在子代稳定遗传表达。在缺铁和铁过量条件下培育T2代转基因植株,缺铁条件下转基因株系抗氧化酶活性(SOD、POD)、叶绿素相对含量、叶片总铁含量和根系生长量均显著高于对照,而丙二醛(MDA)含量显著低于对照;铁过量条件下转基因株系抗氧化酶活性(SOD)、叶片总铁含量、糙米总铁含量和根系生长量均显著高于对照,而MDA含量和稻瘟病叶瘟指数显著低于对照。表明铁蛋白基因NtFer1能够提高转基因植株抗氧化胁迫能力和稻瘟病抗性,增加植株铁离子储藏能力,增强植株缺铁胁迫的耐性和铁过量胁迫的抗性。

In order to realize the gene’s function and improve the resistance ability of japonica,NtFer1 was transformed into japonica Kongyu 131 by Agrobacterium-mediated method. The results by resistance screening,PCR,Southern blotting,Northern blotting,and RT-PCR demonstrated that the gene was integrated into Kongyu 131’s genome,stably and genetically expressed in offspring. T2 generation was cultured in iron deficiency and overload conditions respectively. Under iron deficiency conditions,SOD,POD activity,relative chlorophyll content,total iron content in leaves and roots of transgenic lines were all significantly higher than the control,while the MDA content was lower than the control. Under iron overload conditions,SOD activity,total iron content in leaves and brown rice,and roots growth of transgenic lines were significantly higher than the control,while MDA content and rice blast index were significantly lower than the control. The results revealed that NtFer1 could improve transgenic plant’s ability of oxidative stress and resistance to rice blast,enhance Fe2＋ storage ability and stress resistance in iron deficiency and overload conditions.