水稻D1基因新等位突变体的鉴定与功能分析
收稿日期: 2023-05-06
修回日期: 2023-06-18
网络出版日期: 2024-03-14
基金资助
国家自然科学基金资助项目(32070197);国家级大学生创新创业训练计划资助项目(S202110712667)
Map-based Cloning and Functional Analysis of a New Allele of D1, a Gene Controlling Plant Height in Rice (Oryza sativa L.)
Received date: 2023-05-06
Revised date: 2023-06-18
Online published: 2024-03-14
【目的】株高是作物重要的农艺性状，挖掘株高控制基因并解析其分子功能，可为作物高产育种提供更多有用的基因资源。【方法】利用EMS诱变水稻日本晴获得的矮化突变体d1-11为材料，进行表型和细胞学观察，通过图位克隆的方法鉴定d1-11基因并对基因表达、激素含量和抗旱性进行了分析。【结果】d1-11突变体表现出植株矮化、叶片变短变宽和籽粒形态变圆表型；d1-11突变体叶片中脉萎缩，大脉和小脉数量和面积减少，导致叶片形态变异。d1-11基因被定位到水稻5号染色体R5M15.2和R5M15.8两个分子标记之间；图位克隆结果表明d1-11突变体中D1基因第11外显子和内含子交界处单碱基替换导致基因功能缺失。D1基因在苗期各组织中表达量较高，从分蘖期开始表达量降低；外源脱落酸(ABA)处理24 h后诱导D1基因表达，外源赤霉素(GA)处理抑制D1基因表达，盐胁迫处理24 h诱导D1基因剧烈上调表达。d1-11突变体植株GA、油菜素内酯(BR)和生长素(IAA)等激素含量均上升，叶片相对含水量上升、叶片失水速率降低，植株对干旱胁迫抗性显著增强。【结论】鉴定到水稻D1基因新等位突变d1-11，发现d1-11突变体多种内源激素水平上升、叶片含水量增加、植株对干旱胁迫抗性增强。本研究进一步丰富了水稻矮化基因资源并揭示D1基因新的生物学功能。
高郡茹, 权弘羽, 袁刘珍, 李钦颖, 乔磊, 李文强 . 水稻D1基因新等位突变体的鉴定与功能分析[J]. 中国水稻科学, 2024 , 38(2) : 140 -149 . DOI: 10.16819/j.1001-7216.2024.230502
【Objective】Plant height is an important agronomic trait in rice breeding. Identification of genes controlling plant height and their functional characterization can provide useful genetic resources for high-yield breeding.【Methods】A dwarf mutant, d1-11, was screened from the rice variety Nipponbare by EMS mutagenesis. Phenotypic and cytological observations, gene expression, hormone content and drought resistance were analyzed. The d1-11 mutant gene was identified through map-based cloning.【Results】The d1-11 mutant exhibits dwarfism, with more shortened and widened leaves, and more rounded grains compared to the wild type. The d1-11 mutant leaf has a smaller midvein, reduced number and area of large and small veins, resulting in abnormal leaf morphology in the d1-11 mutant. The d1-11 gene is genetically mapped between two molecular markers on rice chromosome 5. Map-based cloning reveals that a single base substitution at the junction of the ninth exon-intron in the D1 gene results in the loss-of-function mutation of d1-11. The D1 gene has a higher expression level in various tissues at the seedling stage, but the expression levels decrease from the tillering stage. Exogenous abscisic acid (ABA) treatment for 24 hours induces D1 gene expression, exogenous gibberellin (GA) treatment inhibits D1 gene expression, and salt stress treatment for 24 hours can strongly induce D1 gene expression. The contents of several hormones such as GA, brassinosteroid (BR), and indole-3-acetic acid (IAA) were increased in the d1-11 mutant. The d1-11 mutant shows a significant increase in relative water content (RWC) and a reduced rate of water loss in leaves. Furthermore, d1-11 mutant plants exhibit stronger resistance to drought stress.【Conclusion】The d1-11, a novel allele in D1 locus was identified in the present study. It was showed that the d1-11 mutant had increased levels of various endogenous hormones, increased leaf water content, and enhanced resistance to drought stress. This study will further enrich the genetic resources related to dwarfism and reveals some new biological roles of the D1 gene in rice.
Key words： plant height; dwarfism; D1 gene; d1-11 mutant; plant hormones; drought stress
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