KUP/HAK/KT钾转运体基因家族对植物吸收钾离子发挥重要作用,鉴定和克隆棉花的钾转运体基因,对于改良棉花的钾吸收特性,提高棉花的产量和品质具有重要意义。基于已测序的陆地棉基因组序列,本研究通过同源克隆的方法鉴定到陆地棉钾转运体...KUP/HAK/KT钾转运体基因家族对植物吸收钾离子发挥重要作用,鉴定和克隆棉花的钾转运体基因,对于改良棉花的钾吸收特性,提高棉花的产量和品质具有重要意义。基于已测序的陆地棉基因组序列,本研究通过同源克隆的方法鉴定到陆地棉钾转运体基因GhHAK5,并以陆地棉品种百棉1号为材料对其CDS序列进行扩增。结果表明,GhHAK5基因的CDS全长为2451 bp,编码816个氨基酸,分子量和等电点分别为91.23 k D和8.15。GhHAK5蛋白具有KUP/HAK/KT家族基因的保守结构域"K-trans"(Pfam02705)和标志性序列GXXXGDXXXSPLY,并具有11个跨膜区。在进化上,GhHAK5蛋白与拟南芥AtHAK5亲缘关系最近,其次是与水稻的OsHAK5,它们同属Cluster I进化簇。亚细胞定位结果显示,GhHAK5是一个定位于质膜的蛋白,这与其主要作为钾转运子参与K+吸收的功能是一致的。GhHAK5基因在根中表达量最高,在茎、叶、花瓣、纤维和花萼中表达量很低,且其表达受外界低钾环境诱导。本研究结果为进一步了解GhHAK5基因的功能及培育钾高效棉花品种奠定了基础。展开更多
In this study, a gene encoding a superoxide dismutase （SOD） was cloned from senescent leaves of cotton （Gossypium hirsutum）, and its expressing profile was analyzed. The gene was cloned by rapid amplification of c...In this study, a gene encoding a superoxide dismutase （SOD） was cloned from senescent leaves of cotton （Gossypium hirsutum）, and its expressing profile was analyzed. The gene was cloned by rapid amplification of cDNA ends （RACE） method. Northern blotting was used to show the profile of the gene expression, and the enzyme activity was mensurated by NBT deoxidization method in different growth periods. The full length of a gene of cytosolic copper/zinc superoxide dismutase （Cu/Zn-SOD） was isolated from cotton （GenBank Accession Number： DQ445093）. The sequence of cDNA contained 682 bp, the opening reading frame 456 bp, and encoded polypeptide 152 amino acids with the predicted molecular mass of 15.03 kD and theoretical pI of 6.09. The amino acid sequence was similar with the other plants from 82 to 87%. Southern blotting showed that the gene had different number of copies in different cotton species. Northern blotting suggested that the gene had different expression in different tissues and development stages. The enzyme activity was the highest in peak flowering stage. The cotton cytosolic （Cu/Zn-SOD） had lower copies in the upland cotton. The copper/zinc superoxide dismutase mRNA expressing level showed regular changing in the whole development stages; it was lower in the former stages, higher in latter stages and the highest at the peak flowering stage. The curve of the copper/zinc superoxide dismutase mRNA expressing level was consistent with that of the Cu/Zn-SOD enzyme activity. The copper/zinc superoxide dismutase mRNA expressing levels of different organs showed that the gene was higher in the root, leaf, and lower in the flower.展开更多
文摘KUP/HAK/KT钾转运体基因家族对植物吸收钾离子发挥重要作用,鉴定和克隆棉花的钾转运体基因,对于改良棉花的钾吸收特性,提高棉花的产量和品质具有重要意义。基于已测序的陆地棉基因组序列,本研究通过同源克隆的方法鉴定到陆地棉钾转运体基因GhHAK5,并以陆地棉品种百棉1号为材料对其CDS序列进行扩增。结果表明,GhHAK5基因的CDS全长为2451 bp,编码816个氨基酸,分子量和等电点分别为91.23 k D和8.15。GhHAK5蛋白具有KUP/HAK/KT家族基因的保守结构域"K-trans"(Pfam02705)和标志性序列GXXXGDXXXSPLY,并具有11个跨膜区。在进化上,GhHAK5蛋白与拟南芥AtHAK5亲缘关系最近,其次是与水稻的OsHAK5,它们同属Cluster I进化簇。亚细胞定位结果显示,GhHAK5是一个定位于质膜的蛋白,这与其主要作为钾转运子参与K+吸收的功能是一致的。GhHAK5基因在根中表达量最高,在茎、叶、花瓣、纤维和花萼中表达量很低,且其表达受外界低钾环境诱导。本研究结果为进一步了解GhHAK5基因的功能及培育钾高效棉花品种奠定了基础。
文摘In this study, a gene encoding a superoxide dismutase （SOD） was cloned from senescent leaves of cotton （Gossypium hirsutum）, and its expressing profile was analyzed. The gene was cloned by rapid amplification of cDNA ends （RACE） method. Northern blotting was used to show the profile of the gene expression, and the enzyme activity was mensurated by NBT deoxidization method in different growth periods. The full length of a gene of cytosolic copper/zinc superoxide dismutase （Cu/Zn-SOD） was isolated from cotton （GenBank Accession Number： DQ445093）. The sequence of cDNA contained 682 bp, the opening reading frame 456 bp, and encoded polypeptide 152 amino acids with the predicted molecular mass of 15.03 kD and theoretical pI of 6.09. The amino acid sequence was similar with the other plants from 82 to 87%. Southern blotting showed that the gene had different number of copies in different cotton species. Northern blotting suggested that the gene had different expression in different tissues and development stages. The enzyme activity was the highest in peak flowering stage. The cotton cytosolic （Cu/Zn-SOD） had lower copies in the upland cotton. The copper/zinc superoxide dismutase mRNA expressing level showed regular changing in the whole development stages; it was lower in the former stages, higher in latter stages and the highest at the peak flowering stage. The curve of the copper/zinc superoxide dismutase mRNA expressing level was consistent with that of the Cu/Zn-SOD enzyme activity. The copper/zinc superoxide dismutase mRNA expressing levels of different organs showed that the gene was higher in the root, leaf, and lower in the flower.
