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Identification and characterization of the chalkiness endosperm gene CHALK-H in rice(Oryza sativa L.) 被引量:1
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作者 PIAO Ri-hua CHEN Mo-jun +6 位作者 MENG Fan-mei QI Chun-yan KOH Hee-jong GAO Meng-meng SONG An-qi JIN Yong-mei YAN Yong-feng 《Journal of Integrative Agriculture》 SCIE CAS CSCD 2023年第10期2921-2933,共13页
Chalkiness is one of the most important agronomic traits in rice breeding,which directly affects the quality of rice seed.In this study,we identified a chalkiness endosperm mutant,chalk-h,from N-methyl-N-nitrosourea(M... Chalkiness is one of the most important agronomic traits in rice breeding,which directly affects the quality of rice seed.In this study,we identified a chalkiness endosperm mutant,chalk-h,from N-methyl-N-nitrosourea(MNU)-induced japonica rice cultivar Hwacheong(HC).Compared with wild type(WT)-HC,chalk-h showed severe chalkiness in the endosperm,yellowish green leaves,as well as reduced plant height.Scanning electron microscopy(SEM)analysis showed that starch grains in the chalk-h mutant were irregular in size and loosely arranged,with large gaps between granules,forming ovoid or orbicular shapes.MutMap analysis revealed that the phenotype of chalk-h is controlled by a single recessive gene LOC_Os11g39670 encoding seryl-tRNA synthetase,which is renamed as CHALK-H.A point mutation occurs in chalk-h on the sixth exon(at nucleotide 791)of CHALK-H,in which adenine(A)is replaced by thymidine(T),resulting in an amino acid codon change from glutamine(Glu)to valine(Val).The chalk-h mutant exhibited a heat-sensitive phenotype from the 3-leaf stage,including yellow-green leaves and reduced pigment content.The transcriptional expression of starch synthesis-related genes was down-regulated in the chalk-h mutants compared to WT-HC at different grain-filling stages.With an increase in temperature,the expression of photosynthesis-related genes was down-regulated in the chalk-h mutant compared to WT-HC.Overexpression of CHALK-H rescued the phenotype of chalk-h,with endosperm and leaf color similar to those of WT-HC.Our findings reveal that CHALK-H is a causative gene controlling chalkiness and leaf color of the chalk-h mutant.CHALK-H is the same gene locus as TSCD11,which was reported to be involved in chloroplast development under high temperature.We suggest that CHALK-H/TSCD11 plays important roles not only in chloroplast development,but also in photosynthesis and starch synthesis during rice growth and development,so it has great application potential in rice breeding for high quality and yield. 展开更多
关键词 CHALK-H seryl-trna synthetase CHALKINESS RICE
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丝氨酰tRNA合成酶(SerRS)结合并激活去乙酰化酶SIRT2的机制
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作者 秦懿偲 周德健 +1 位作者 李超群 余巍 《复旦学报(医学版)》 CAS CSCD 北大核心 2020年第4期506-512,530,共8页
目的探究丝氨酰tRNA合成酶(seryl-tRNA synthetase,SerRS)结合并激活去乙酰化酶SIRT2的机制。方法构建SerRS与SIRT2质粒,在293T细胞中进行转染。免疫共沉淀方法检测SerRS与SIRT2、SIRT2突变型之间的相互作用。通过原核纯化蛋白质,体外... 目的探究丝氨酰tRNA合成酶(seryl-tRNA synthetase,SerRS)结合并激活去乙酰化酶SIRT2的机制。方法构建SerRS与SIRT2质粒,在293T细胞中进行转染。免疫共沉淀方法检测SerRS与SIRT2、SIRT2突变型之间的相互作用。通过原核纯化蛋白质,体外荧光方法检测SerRS对SIRT2酶活影响。通过结构模拟探究SIRT2的H187位点调控机制。通过Western blot方法检测SerRS的乙酰化修饰。结果SerRS能与SIRT2相互作用,并增强其去乙酰化酶活性。SIRT2 H187Y突变使SIRT2失活,同时也阻断了SIRT2与其SerRS的结合。进一步通过结构模拟发现,SIRT2上的H187Y突变与Q267位点在空间上发生冲突,可能改变SIRT2的构象,进而影响与SerRS的结合界面,且SIRT2能够去乙酰化SerRS。结论SIRT2第187位组氨酸能够调控SerRS与SIRT2的结合,为靶向SIRT2的药物研发提供了新的思路。 展开更多
关键词 乙酰化 丝氨酰tRNA合成酶(SerRS) SIRT2 血管生成
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