水稻裂颖突变体sh1的鉴定及基因定位

doi:10.16819/j.1001-7216.2019.9027

摘要/Abstract

摘要：

【目的】本研究旨在定位和克隆水稻裂颖基因,为解析水稻裂颖性的遗传机制提供依据。【方法】从籼稻品种湘早籼6号突变体库中筛选出一个裂颖突变体(split husk 1, sh1),观察突变体的花器官和浆片形态,利用突变体与02428的F₂群体定位目标基因,进一步通过定量PCR分析相关基因的表达情况。【结果】sh1突变体的颖花形态与野生型基本一致,能正常开花,但不能正常闭颖,裂颖的主要原因是浆片不能在开颖后正常萎蔫。sh1突变体的有效穗数增加,但结实率和千粒重显著下降;遗传分析表明,sh1的裂颖表型受一对隐性核基因控制。将SH1基因定位在ID19827与ID19884两个InDel标记之间,物理距离约为110 kb。定位区间测序发现,突变体中丙二烯氧化合酶编码基因OsAOS1发生单碱基突变,导致氨基酸发生改变;SH1基因的突变显著降低了花器官中的茉莉酸含量,进而影响了茉莉酸合成及信号转导相关基因的表达。【结论】SH1基因通过影响茉莉酸的合成和信号转导调控水稻闭颖,OsAOS1可能是SH1基因的候选基因。

关键词: 水稻, 颖花闭合, 浆片, 基因定位, 茉莉酸

Abstract:

【Objective】The objective of the research is to identify and clone a rice split-husk gene, providing support for elucidating the genetic mechanism of rice floret closing. 【Method】A split husk mutant, sh1, was isolated from an ethylmethylsulfone (EMS) mutagenic population of indica cultivar Xiangzaoxian 6. The morphological characteristics of florets and lodicules were observed. The SH1 gene was identified by map-based cloning using an F₂ population of a cross between sh1 and 02428. In addition, expression levels of related genes were analyzed by quantitative PCR. 【Results】The sh1 mutant showed normal floral morphological characteristics, but failed to close the lemma and palea after floret opening, which was caused by delay of lodicule dehydration. The sh1 mutant was also featured by increased number of effective panicles, obviously decreased seed-setting rate and 1000-grain weight. Genetic analysis indicated that the mutant phenotype was controlled by a pair of recessive nuclear gene. The SH1 gene was fine-mapped to a 110 kb interval between markers ID19827 and ID19884 on the chromosome 3. After sequencing of this region, we found that there is a single base transition in the coding region of allene oxide synthase 1 (OsAOS1), which resulted in an amino acid substitution. Correspondingly, the mutant showed reduced content of jasmonic acid (JA) and differential expression of genes related to JA synthesis and signal transduction. 【Conclusion】The SH1 gene regulates rice floret closing through JA synthesis and signal transduction, and OsAOS1 is the candidate gene for SH1.

Key words: rice, floret closing, lodicule, gene mapping, jasmonate

中图分类号:

潘孝武, 刘文强, 黎用朝, 熊海波, 盛新年, 段永红, 余亚莹, 赵文锦, 魏秀彩, 李小湘. 水稻裂颖突变体sh1的鉴定及基因定位[J]. 中国水稻科学, 2019, 33(4): 323-330.

Xiaowu PAN, Wenqiang LIU, Yongchao LI, Haibo XIONG, Xinnian SHENG, Yonghong DUAN, Yaying YU, Wenjin ZHAO, Xiucai WEI, Xiaoxiang LI. Identification and Genetic Analysis of Split Husk Mutant sh1 in Rice[J]. Chinese Journal OF Rice Science, 2019, 33(4): 323-330.

图/表 6

表1 基因定位及表达分析引物信息

Table 1 Primers for gene mapping and expression analysis.

引物名称Primer name	正向引物 Forward primer (5'→3')	反向引物 Reverse primer (5'→3')
RM15967	AAGAGCCCTAGCGATAGCAAAGC	GTTCTACCTACCTGCGGCTTCC
RM1352	CACCTTCGATTTCCTTCTCATCG	CGAGTTGTACTCTGGTTGCTTCC
RM1004	GTCGACATGCTCGTGGTTCTGG	CCCAAGTTCGTCGACCTCACC
ID19827	ACCGTTCACTACAAGCAGAT	GTGTTCCACTTCCTCAACTG
ID19884	AAGTGGATCTCTCCGAAACT	TTCTGAGGGAGCTTATCGTA
OsActin	CATTGGTGCTGAGCGTTTCC	AGAAACAAGCAGGAGGACGG
OsJAmyb	GAGGACCAGAGTGCAAAAGC	CATGGCATCCTTGAACCTCT

图1 野生型和突变体颖花的形态学观察

Fig. 1. Morphological observation of florets of wild type and sh1 mutant.

表2 野生型与突变体sh1的主要农艺性状比较

Table 2 Agronomic traits of the wild type (WT) and sh1 mutant.

材料 Material	株高 Plant height / cm	有效穗数 No. of effective panicles	每穗总粒数 No. of spikelets per panicle	结实率 Seed-setting rate / %	千粒重 1000-grain weight / g
WT	81.4±3.4	5.5±0.5	134.3±6.7	81.8±6.3	23.9±0.4
sh1	87.8±2.1	7.5±0.7^*	121.3±12.9	25.1±3.8^**	22.1±0.3^*

图2 水稻SH1基因的精细定位

Fig. 2. Fine mapping of the rice SH1 gene.

图3 野生型和突变体花器官中的茉莉酸含量**表示突变体与野生型之间的差异达0.01显著水平。

Fig. 3. JA content in florets of wild type and sh1 mutant. ** Significant difference between the mutant and WT at 0.01 level.

图4 茉莉酸合成及信号转导相关基因的相对表达量*表示突变体与野生型之间的差异达0.05显著水平;**表示突变体与野生型之间的差异达0.01显著水平。

Fig. 4. Relative expression level of genes related to JA synthesis and signal transduction. *Significant difference between the mutant and WT at 0.05 level; **Significant difference between the mutant and WT at 0.01 level.

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