P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phy...P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.展开更多
【目的】叶色突变体是研究水稻光合作用,叶绿素生物合成和遗传发育调控机理的重要材料。发掘水稻叶色突变体,是水稻功能基因组学研究的重要遗传基础。【方法】在昌恢121中发现了一份白条纹叶及抽穗期白穗突变体,经过连续多代自交能稳定...【目的】叶色突变体是研究水稻光合作用,叶绿素生物合成和遗传发育调控机理的重要材料。发掘水稻叶色突变体,是水稻功能基因组学研究的重要遗传基础。【方法】在昌恢121中发现了一份白条纹叶及抽穗期白穗突变体,经过连续多代自交能稳定遗传,暂命名为wlp6(white striped leaf and white panicle 6)。在南昌分早、中和晚3季播种wlp6与野生型种子,考查了中稻与晚稻的部分农艺性状;测定3叶期、分蘖期、抽穗期叶片及颖壳的叶绿素含量;通过电镜观察抽穗期叶肉细胞发育情况。在光照培养箱中进行温光敏感实验;将wlp6与昌恢121及02428正反交,观察F_1植株表型,对F_2分离群体进行卡方测验,分析突变体遗传规律;以wlp6/02428衍生的F_2群体为材料,利用BSA法进行基因定位。【结果】wlp6自第1片叶到成熟,叶片均呈白条纹,抽穗期颖壳及枝梗失绿,高温天气穗转绿。突变体株高、有效穗数和每穗粒数在早稻季和中稻季均显著低于野生型,晚稻季wlp6的结实率和千粒重也显著低降低。叶绿素含量测定表明,wlp6叶片叶绿素含量在不同生育期及不同季均显著低于野生型,早稻和晚稻季种植的wlp6颖壳叶绿素含量也比野生型低。电镜观察抽穗期的叶肉细胞发现,wlp6叶绿体数目减少,体积变小,没有分化出明显的片层结构。温光敏感实验表明,突变体对光照强弱钝感,叶色受温度和日照长短影响,随着温度升高和日照时间变长突变体叶绿素含量有上升趋势。遗传分析表明,该性状受隐性核基因控制,利用wlp6/02428得到的616个F_2单株将WLP6定位于第6染色体短臂InDel标记R-7与R-8间,物理距离137kb,此区间预测了21个候选基因。经候选基因分析及测序发现,其中LOC_Os06g14620编码一个核糖核酸还原酶小亚基,编码区第142和158位碱基由T替换为C,第288位插入了碱基A,碱基的插入导致翻译提前终止,因此推测LOC_Os06g14620是WLP6的候选基因。【结论】LOC_Os06g14620是已经克隆的白条纹叶基因St1的候选基因,推测WLP6与St1等位,但突变位点不同,且表型也有差异。展开更多
基金the Agricultural Science and Technology Innovation Program Cooperation and Innovation Mission(Grant No.CAAS-XTCX2016001)Shenzhen Science and Technology Projects(Grant No.JSGG20160608160725473)+1 种基金China Postdoctoral Science Foundation(Grant No.2018M641558)Fundamental Research Funds for Science,Technology and Innovation Commission of Shenzhen Municipality(Grant No.JCYJ20160530191619099).
文摘P1B-type heavy metal ATPases(HMAs)are transmembrane metal-transporting proteins that play a key role in metal homeostasis.We here reported the characterization of rice OsHMA6,a member of the P1B-type ATPase family.Phylogenetic tree analysis showed that OsHMA6 belonged to the Cu/Ag subgroup of the HMA family and had a close evolutionary relationship with OsHMA9.Amino acid sequence alignment showed 82.78%consistency between OsHMA6 and OsHMA9.OsHMA6 expressed in all organs at different growth stages,including spikelet,and abundant in leaf blades,however,OsHMA9 most strongly expressed in roots,but very low in spikelet.Excessive Cu^2+can up-regulate the expression of OsHMA6 and OsHMA9 in rice seedlings.The heterologous expression in yeast showed that OsHMA6 can significantly rescue the growth of yeast strain CM52 when supplied with 3 or 6 mmol/L Cu^2+.Compared with the empty vector pYES2,the Cu concentration in OsHMA6-pYES2 decreased by 23.4%and 30.3%under 3 or 6 mmol/L Cu2+,respectively.Subcellular localization revealed that OsHMA6 was located in the plasma membrane.These results suggested that OsHMA6,similar to OsHMA9,is likely a copper efflux protein located in the plasma membrane.
文摘【目的】叶色突变体是研究水稻光合作用,叶绿素生物合成和遗传发育调控机理的重要材料。发掘水稻叶色突变体,是水稻功能基因组学研究的重要遗传基础。【方法】在昌恢121中发现了一份白条纹叶及抽穗期白穗突变体,经过连续多代自交能稳定遗传,暂命名为wlp6(white striped leaf and white panicle 6)。在南昌分早、中和晚3季播种wlp6与野生型种子,考查了中稻与晚稻的部分农艺性状;测定3叶期、分蘖期、抽穗期叶片及颖壳的叶绿素含量;通过电镜观察抽穗期叶肉细胞发育情况。在光照培养箱中进行温光敏感实验;将wlp6与昌恢121及02428正反交,观察F_1植株表型,对F_2分离群体进行卡方测验,分析突变体遗传规律;以wlp6/02428衍生的F_2群体为材料,利用BSA法进行基因定位。【结果】wlp6自第1片叶到成熟,叶片均呈白条纹,抽穗期颖壳及枝梗失绿,高温天气穗转绿。突变体株高、有效穗数和每穗粒数在早稻季和中稻季均显著低于野生型,晚稻季wlp6的结实率和千粒重也显著低降低。叶绿素含量测定表明,wlp6叶片叶绿素含量在不同生育期及不同季均显著低于野生型,早稻和晚稻季种植的wlp6颖壳叶绿素含量也比野生型低。电镜观察抽穗期的叶肉细胞发现,wlp6叶绿体数目减少,体积变小,没有分化出明显的片层结构。温光敏感实验表明,突变体对光照强弱钝感,叶色受温度和日照长短影响,随着温度升高和日照时间变长突变体叶绿素含量有上升趋势。遗传分析表明,该性状受隐性核基因控制,利用wlp6/02428得到的616个F_2单株将WLP6定位于第6染色体短臂InDel标记R-7与R-8间,物理距离137kb,此区间预测了21个候选基因。经候选基因分析及测序发现,其中LOC_Os06g14620编码一个核糖核酸还原酶小亚基,编码区第142和158位碱基由T替换为C,第288位插入了碱基A,碱基的插入导致翻译提前终止,因此推测LOC_Os06g14620是WLP6的候选基因。【结论】LOC_Os06g14620是已经克隆的白条纹叶基因St1的候选基因,推测WLP6与St1等位,但突变位点不同,且表型也有差异。
