拟南芥AtmiR399f基因在番茄中异位表达对低磷耐性及外观形态的影响  

Effect of Ectopic Expression of Arabidopsis AtmiR399f Gene on Tolerance to Phosphorus Deficiency and Appearance Morphology in Tomato

作　　者：赵先炎 刘泽洲[1] 王双双[1] 陈可钦[1] 郝玉金[1] 由春香[1] 

机构地区：[1]作物生物学国家重点实验室,国家苹果工程技术研究中心,山东农业大学园艺科学与工程学院,泰安271018

出　　处：《西北农业学报》2015年第3期129-136,共8页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：国家自然科学基金(31171946);教育部"长江学者和创新团队发展计划"创新团队(IRT1155)

摘　　要：为了研究miR399过量表达的番茄对低磷胁迫的响应,将拟南芥AtmiR399f基因转入番茄中,获得超表达量不同的2个株系,探讨转基因番茄外观形态表现以及耐低磷的可能机制。构建植物表达载体pBI121-AtmiR399f,用农杆菌介导法将拟南芥AtmiR399f基因转化到番茄体内,用茎环半定量RT-PCR法进一步确定2株不同表达水平的转AtmiR399f基因番茄株系,以此为试验材料,用茎环反转录实时荧光定量PCR检测miR399的表达量,用不同浓度磷处理株系,并且实时荧光定量PCR检测与磷运输有关的基因的表达量以及半定量RT-PCR检测与乙烯信号途径有关的基因的表达量。与野生型番茄株系相比,转基因番茄产生更多的侧根及次级侧根,且种子更大、茎秆更粗壮,对低磷逆境环境耐性更强。另外,磷运输相关基因LePT1没变化,乙烯合成途径中pTOM13基因和ACS基因表达量也升高。异位表达AtmiR399f基因的转基因番茄株系miR399表达量增高,对低磷耐性增强,miR399是通过调控乙烯合成途径来影响植物对磷的吸收。For studying response of over-expressed miR399 fin tomato to stress of phosphorus deficiency,two different tomato lines were obtained by transferring arabidopsis AtmiR399 fgene into tomatos,the possible mechanism of low-phosphorus tolerance;and appearance morphology in transgenic tomato was discussed in the paper.pBI121-AtmiR399 fvector was structured and introduced into tomato by Agrobacterium-mediated transformation.Two transgenic lines with different expression levels of AtmiR399 fgene were confirmed by stem-loop RT-PCR technique to characterize the function of miR399 in tomato,and then to detect the the expression of miR399 by stem-loop RT-PCR technique and the expression of genes related to the phosphorus transport volume by real-time quantitative PCR and the expression of genes associated with ethylene signaling pathways with RT-PCR;the results showed that these two transgenic tomatoes had changed the root architecture and produced a highlybranched-root system with abundant lateral and secondary lateral roots,which could enhance lowphosphorus tolerance.Futhermore,these tomatoes produced larger seeds and increased the stem diameter.In addition,LePT1 gene related to the phosphorus transport remained unchanged,but the expression of pTOM13 and ACSgenes associated with ethylene signaling pathways increased;the findings indicated that overexpression of miR399 with ectopically overexpressed AtmiR399 fgene made the transgenic tomato.The phosphorus resistance increased by mediating the ethylene synthesis.

关 键 词：番茄 拟南芥AtmiR399f基因 磷 乙烯 MIRNAS 

分 类 号：Q943.2[生物学—植物学]