水稻矮杆突变体的细胞学特征及基因定位研究

郝小花, 向玉婷, 曾 孟, 杨友伟, 谢 鹏, 李 密, 李东屏, 田连福

生命科学研究 ›› 2021, Vol. 25 ›› Issue (1) : 39-47.

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PDF(12358 KB)
生命科学研究 ›› 2021, Vol. 25 ›› Issue (1) : 39-47.
经济动植物

水稻矮杆突变体的细胞学特征及基因定位研究

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Cytological Characteristics and Gene Location of a Rice Dwarf Mutant

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摘要

摘 要: 赤霉素合成基因在植物的生长发育中起着重要的作用。甲基磺酸乙酯(ethyl methane sulfonate, EMS)诱变的超矮杆突变株水稻ag1 (ai-gan1)表现出明显的节间和叶片伸长缺陷, 成熟期株高仅为野生型的34.81%, 穗长以及各节间长度占整株高度的比例发生明显变化, 其中倒二节间和倒三节间所占比例显著降低。MBS (mapping-by-sequencing)基因定位及CRISPR-Cas9基因敲除验证显示, 该突变基因为赤霉素-3β羟化酶2 (gib- berellin 3-beta-hydroxylase 2, GA3ox2)编码基因D18的等位基因。细胞的形态分析显示, 突变体倒二节间的细胞长度显著变短, 纵向细胞数量显著减少, 而节间中横向薄壁细胞层数显著增加; 突变体剑叶中横向小叶脉的数量显著增加, 但维管束周围的纵向薄壁细胞数目显著减少。同时, 部分与细胞分裂和伸长相关基因的表达量在突变体中都显著降低。上述结果说明GA3ox2在水稻中严重影响地上节间细胞的伸长及节间和剑叶中细胞的分裂活性, 从而导致水稻地上部分的形态出现异常。

Abstract

Abstract: Gibberellin synthesis genes play an important role in plant growth and development. The super dwarf mutant ai-gan1 (ag1) induced by ethyl methane sulfonate (EMS) showed obvious defects in internode and leaf elongation. The plant height at mature stage reached only 34.81% of the wild-type (WT) height. The panicle length and the ratio of internode length to the whole plant height changed significantly. The proportion of the last second and third internodes had a significant decrease in the total length. Mapping-by-sequencing (MBS) and CRISPR-Cas9 mediated gene knockout confirmed that the mutant gene was an allele encoding gibberellin 3-beta-hydroxylase 2 (GA3ox2). Morphological analysis showed that the length of inter-node cells in the penultimate internode of the mutants was significantly shorter, and the number of longitudinal cells was significantly reduced. The number of transverse parenchyma cells in internodes increased significantly, and the number of transverse venules in flag leaves of the mutants increased significantly, but the number of longitudinal parenchyma cells around vascular bundles decreased significantly. At the same time, the expression levels of some genes related to cell division and elongation were significantly decreased in the mutants. The results showed that GA3ox2 in rice seriously affected the elongation of aboveground internode cells and the cell division activity in internodes and flag leaves, which led to the abnormal morpho-logy of the aboveground part of rice.

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郝小花, 向玉婷, 曾 孟, . 水稻矮杆突变体的细胞学特征及基因定位研究[J]. 生命科学研究, 2021, 25(1): 39-47
HAO Xiao-hua, XIANG Yu-ting, ZENG Meng, et al. Cytological Characteristics and Gene Location of a Rice Dwarf Mutant[J]. Life Science Research, 2021, 25(1): 39-47

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