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玉米漆酶ZmLac5基因拟南芥遗转转化
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作者 莫乔程 赵永平 +2 位作者 朱苏文 程备久 谢传晓 《安徽农业大学学报》 CAS CSCD 北大核心 2012年第4期533-539,共7页
旨在模式生物拟南芥中获得玉米漆酶ZmLac5基因过表达转化体,以研究该基因功能。分离玉米自交系掖478根部总RNA,反转录PCR法获得玉米漆酶基因ZmLac5全长,测序与序列分析表明,ZmLac5 cDNA的开放阅读框为1 758 bp,编码一个由586个氨基酸组... 旨在模式生物拟南芥中获得玉米漆酶ZmLac5基因过表达转化体,以研究该基因功能。分离玉米自交系掖478根部总RNA,反转录PCR法获得玉米漆酶基因ZmLac5全长,测序与序列分析表明,ZmLac5 cDNA的开放阅读框为1 758 bp,编码一个由586个氨基酸组成的蛋白质。同源性比对和进化树分析显示该基因与高粱Sb03g039570.1和谷子Si000813m在进化上亲缘关系较近。采用In-Fusion试剂盒构建了过表达载体pCUB-Ubi::ZmLac5,通过电击法将过表达载体导入根癌农杆菌LBA4404中,并成功转化拟南芥(Columbia ecotype),获得PCR阳性T1代植株62株,转化率为1.28%,T2代阳性植株60株。本研究为基于模式生物体过表达分析该基因生物学功能奠定了分析基础。 展开更多
关键词 玉米 漆酶 ZmLac5 过表达 遗传转化
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MicroRNA528 Affects Lodging Resistance of Maize by Regulating Lignin Biosynthesis under Nitrogen-Luxury Conditions 被引量:32
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作者 Qing Sun Xiaogang Liu +5 位作者 Juan Yang Wenwen Liu Qingguo Du Hongqiu Wang Chunxiang Fu Wen-Xue Li 《Molecular Plant》 SCIE CAS CSCD 2018年第6期806-814,共9页
Lodging under nitrogen (N)-Iuxury conditions substantially reduces crop yield and seed quality. However, the molecular mechanisms of plant lodging resistance remain largely unclear, especially in maize. We report he... Lodging under nitrogen (N)-Iuxury conditions substantially reduces crop yield and seed quality. However, the molecular mechanisms of plant lodging resistance remain largely unclear, especially in maize. We report here that the expression of ZmmiR528, a monocot-specific microRNA, is induced by N luxury but reduced by N deficiency. We show by the thioacidolysis and acetyl bromide analysis that N luxury signifi- cantly reduces the generation of H, G, and S monomers of the lignin as well as its total content in maize shoots. We further demonstrate that ZmLACCASE3 (ZmLAC3) and ZmLACCASE5 (ZmLAC5), which encode the copper-containing laccases, are the targets of ZmmiR528. In situ hybridization showed that ZmmiR528 is mainly expressed in maize vascular tissues. Knockdown of ZmmiR528 or overexpression of ZmLAC3 significantly increased the lignin content and rind penetrometer resistance of maize stems. In contrast, transgenic maize plants overexpressing ZmmiR528 had reduced lignin content and rind penetrometer resistance and were prone to lodging under N-luxury conditions. RNA-sequencing analysis revealed that ZmPAL7 and ZmPAL8 are upregulated in transgenic maize lines downregulating ZmmiR528. Under N-lux- ury conditions, the expression levels of ZmPALs were much higher in ZmmiR528-knockdown lines than in the wild type and transgenic maize lines overexpressing ZmmiR528. Taken together, these results indicate that, by regulating the expression of ZmLAC3 and ZmLAC5, ZmmiR528 affects maize lodging resistance un- der N-luxury conditions. 展开更多
关键词 MAIZE NITROGEN ZmmiR528 zmlacs lodging resistance
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