水稻垩白调控基因CHALK-H的鉴定及特性分析

doi:10.1016/j.jia.2023.04.020

摘要/Abstract

摘要：

垩白是水稻育种中最重要的农艺性状之一，直接影响稻米的品质。本研究利用化学诱变剂MNU处理韩国粳稻品种Hwacheong（HC），获得一个稳定遗传的垩白突变体chalk-h。与野生型HC相比，chalk-h突变体籽粒表现出垩白特性，叶片呈黄绿色，株高降低。扫描电镜(SEM)分析表明，chalk-h突变体淀粉粒大小不规则、排列松散、颗粒间间隙大、呈椭圆形或圆形。MutMap分析显示，chalk-h突变体的垩白表型由编码seryl-tRNA合成酶的单隐性基因LOC_Os11g39670控制，该基因被重新命名为CHALK-H。在chalk-h突变体中CHALK-H基因的第6外显子(第791核苷酸)发生点突变，其腺嘌呤(A)被胸腺嘧啶(T)取代，导致氨基酸密码子由谷氨酰胺(Glu)变为缬氨酸(Val)。chalk-h突变体从三叶期开始表现为黄绿色叶片、色素含量降低等热敏表型。与野生型HC相比，chalk-h突变体在不同灌浆期淀粉合成相关基因的转录表达下调，随着温度的升高，chalk-h突变体中光合相关基因的表达下调。将CHALK-H基因过量表达于chalk-h突变体，其垩白胚乳和黄绿叶片颜色恢复为野生型。研究结果表明，CHALK-H控制稻米垩白和叶片颜色。CHALK-H是已报道参与高温下叶绿体发育相关基因TSCD11的等位基因。我们认为，CHALK-H/TSCD11在水稻生长发育过程中不仅参与叶绿体发育，而且参与光合作用和淀粉合成，在水稻优质高产育种中具有广阔的应用前景。

Abstract:

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.

Key words: CHALK-H , Seryl-tRNA synthetase , chalkiness , rice

PIAO Ri-hua, CHEN Mo-jun, MENG Fan-mei, QI Chun-yan, KOH Hee-jong, GAO Meng-meng, SONG An-qi, JIN Yong-mei, YAN Yong-feng.

水稻垩白调控基因CHALK-H的鉴定及特性分析 [J]. Journal of Integrative Agriculture, 2023, 22(10): 2921-2933.

PIAO Ri-hua, CHEN Mo-jun, MENG Fan-mei, QI Chun-yan, KOH Hee-Jong, GAO Meng-meng, SONG An-qi, JIN Yong-mei, YAN Yong-feng. Identification and characterization of the chalkiness endosperm gene CHALK-H in rice (Oryza sativa L.)[J]. Journal of Integrative Agriculture, 2023, 22(10): 2921-2933.

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