水稻白化转绿和穗顶端退化突变体vpa1的遗传分析和基因定位

doi:10.16819/j.1001-7216.2021.0702

摘要/Abstract

摘要： 目的通过对水稻转绿和穗顶端退化等突变体的研究,可以鉴定更多与叶绿体发育和穗发育相关的基因。方法在常规种植条件下比较突变体vpa1（virescent and panicle abortion 1)表型及主要农艺性状差异,利用分离群体分析和图位克隆法进行相关基因定位。结果突变体vpa1表现苗期白化,并逐渐转绿恢复成正常叶色,抽穗后可明显观查到穗顶端退化表型。vpa1的主要农艺性状除了结实率以外,株高、穗长、每穗实粒数等均较野生型显著下降。遗传分析表明白化转绿和穗顶端退化表型受独立的两个隐性基因控制。控制白化转绿叶性状的Osv16定位于第3染色体RM3441和RM3029之间约125kb物理区间内,区间内未见白化转绿性状相关基因的报道。控制穗顶端退化性状的Ospaa10定位于第1染色体RM11157和RM5972之间,区间内物理距离约190kb,区间内未见穗顶端退化相关基因的报道。结论 Osv16和Ospaa10两个基因的突变导致vpa1的叶色和穗型同时出现变异,为白化转绿基因Osv16和穗顶端退化基因Ospaa10的克隆和功能研究打下了基础。

关键词: 水稻, 转绿, 顶穗退化, 基因定位

Abstract:

【Objective】Genes related to chloroplast development and panicle phenotype can be identified through the study onvirescent mutants and panicle apical abortion (paa) mutants in rice. 【Method】The main agronomic traits of vpa1 (virescent and panicle abortion 1) mutant and the wild-type were compared under conventional planting conditions in field. The virescent gene and paagene were mapped by bulked segregant analysis and map-based cloning.【Result】vpa1exhibited albino leaves at seedling stage andthe leaves graduallyreturned to normal. During heading stage,vpa1showed degeneration of spikelets at the tops of panicles. Except seed-setting rate, the agronomic traits including plant height, panicle length, number of filled grains per panicle and 1000-grain weight decreased significantly in vpa1as compared with the wild-type.Genetic analysis indicated that both the virescentphenotypeand the paa phenotype of vpa1are controlled by single recessive nuclear gene, and the virescentgene and paa gene were segregated independently.The virescent gene, Osv16, was mapped to a 125kb region between RM3441 and RM3029 on chromosome 3. Thepaagene, Ospaa10, was mapped to a 190kb region between RM11157 and RM5972 on chromosome 1.【Conclusion】mutations ofOsv16 and Ospaa10 causedsimultaneous variation of leaf color and spikelet phenotype in vpa1. Osv16 and Ospaa10are likely two new genes as no other virescent and paa genes have been identified in the related region, further cloning of both genes are currently underway.

Key words: rice, virescent, panicle apical abortion, gene mapping

王中豪, 贺彦, 张晓波, 徐霞, 吴建利, 施勇烽. 水稻白化转绿和穗顶端退化突变体vpa1的遗传分析和基因定位[J]. 中国水稻科学, 2021, 35(1): 19-26.

Zhonghao WANG, Yan HE, Xiaobo ZHANG, Xia XU, WUJianli, SHIYongfeng. Genetic Analysis and Gene Mapping of a Virescent and Panicle AbortionMutant vpa1in Rice[J]. Chinese Journal OF Rice Science, 2021, 35(1): 19-26.

图/表 7

图1 突变体vpa1及其野生型在苗期和抽穗期的表型 A-播种10 d后vpa1和野生型IR64的表型,标尺为2 cm;B-vpa1和野生型IR64在抽穗期的表型,标尺为20 cm;C-vpa1和野生型剑叶表型比较,标尺为2 cm;D-vpa1和野生型IR64的穗部表型比较,标尺为4 cm。

Fig. 1. Performance of vpa1 and wild type at the seedling and heading stages. A,vpa1 and its wild type(WT) at 10 days after sowing. Scale bar=2 cm; B,vpa1 and WT at the heading stage. Scale bar=20 cm; C,Flag leavesofvpa1 and WT. Scale bar=2cm; D,Paniclesofvpa1 and WT. Scale bar= 4cm.

表1 突变体vpa1和野生型IR64的主要农艺性状

Table l. Comparison of agronomnic traits between vpa1 and the wild-type.

材料 Material	株高 Plant height/cm	穗长 Panicle length /cm	有效穗数 No.of tillers per plant	每穗实粒数 No.of filledgrains per panicle	结实率 Seed-setting rate / %	千粒重 1000-grain weight / g
WT	107.7±2.1	26.0±1.0	17.7±0.6	84.9±2.3	61.6±0.9	27.0±0.1
vpa1	91.7±1.5^**	18.8±0.8^**	13.3±2.1^*	58.3±2.2^**	62.6±3.3	21.4±0.4^**

图2 不同时期vpa1和野生型IR64叶片光合色素含量的比较 A-播种后2周vpa1和野生型IR64的色素含量;B-播种后8周vpa1和野生型IR64的色素含量;C-播种后14周vpa1和野生型IR64的色素含量;误差线表示3次生物学重复试验的标准差,*和**分别表示野生型和vpa1之间差异达0.05和0.01显著水平（t检验）。

Fig. 2. Comparison of pigment contents between vpa1 and WT at different growth stages. A, Pigment contents of vpa1 and WT at two weeks after sowing; B, Pigment contents of vpa1 and WT at eight weeks after sowing; C, Pigment contents of vpa1 and WT at 14 weeks after sowing; The values are means ± SD (n = 3). The asterisks indicate significant difference between vpa1 and WT according to the Student’s t-test(*P≤0.05; **P≤0.01).

表2 vpa1中转绿叶性状的遗传分析

Table 2 Genetic analysis of the virescent leaf in vpa1.

杂交组合Cross	F₁	F₂			χ²_3:1
杂交组合Cross	F₁	正常叶色株数 No.of normal plants	白化转绿株数 No.of virescent plants	总株数 Total no. of plants	χ²_3:1
vpa1/IR64	正常叶色Green	726	254	980	0.44
vpa1/Moroberekan	正常叶色Green	3892	1372	5264	3.18

表3 vpa1中穗顶端退化性状的遗传分析

Table 3 Genetic analysis of the paatrait in vpa1.

杂交组合 Cross	F₁	F₂			χ²_3:1
杂交组合 Cross	F₁	正常穗型株数 No.of normal panicle plants	顶端退化株数 No. of paaplants	总株数 Total no.of plants	χ²_3:1
vpa1/IR64	正常穗型Normal panicle	565	155	720	3.58
vpa1/Moroberekan	正常穗型Normal panicle	1181	254	1435	40.78

图3 白化转绿基因Osv16在第3染色体上的定位 A-白化转绿基因Osv16的初步定位;B,C-白化转绿基因Osv16的精细定位。

Fig. 3. Location of Osv16 on chromosome 3. A, Preliminary localization of Osv16 gene; B andC, Fine mapping of Osv16 gene.

图4 穗顶端退化基因Ospaa10在第1染色体上的定位 A-穗顶端退化基因Ospaa10的初步定位;B, C-穗顶端退化基因Ospaa10的精细定位。

Fig. 4. Location of Ospaa10 on chromosome 1. A, Preliminary localization of Ospaa10 gene; B and C, Fine mapping of Ospaa10 gene.

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