Genetic Analysis and Gene Mapping of a Heading-delayed Mutant dth9 in Rice (Oryza sativa L.)

doi:10.16819/j.1001-7216.2016.5156

Abstract

Abstract:

A heading-delayed mutant, dth9 (days to heading 9)was identified from an ethyl methylsulfonate (EMS)-induced 93-11 mutant library. The heading date of dth9 delayed about 50 days compared to the wild-type and there was no significant difference in other agronomic traits. Genetic analysis showed that the phenotype of dth9 was controlled by a single recessive nuclear gene. To map this gene, two F₂populations were generated by crossing the dth9 mutant with Nipponbare or Wuyunjing 7 as mapping populations. By using SSR markers and eight new designed InDel markers, DTH9 was narrowed to a 240kb interval between the markers D9-9 and D9-17 near the centromere of chromosome 9, there were no reports about genes associated with heading date in this interval. In addition, the expression levels of genes related to heading date were significantly decreased in dth9 by quantitative real-time PCR analysis.

Key words: rice, heading date, genetic analysis, gene mapping

摘要：

在EMS诱变的93-11突变体库中筛选到一个稳定遗传的迟抽穗突变体dth9 (days to heading 9)。该突变体的抽穗期比野生型延长了50d左右,其他农艺性状基本无异。遗传分析表明迟抽穗性状受一个隐性核基因控制。以突变体dth9与日本晴和武运粳7号杂交构建的F₂分离群体作为定位群体,利用SSR标记和新开发的8个InDel标记,将DTH9定位在第9染色体着丝粒附近D9-9和D9-17之间240 kb的区间内,该区域尚未发现与抽穗期有关的基因。此外,实时荧光定量PCR结果表明,在突变体dth9中与抽穗期相关基因的表达量显著降低。

关键词: 水稻, 抽穗期, 遗传分析, 基因定位

CLC Number:

Wei-jun YE, Shi-kai HU, Li-wen WU, Long-biao GUO, Qian QIAN. Genetic Analysis and Gene Mapping of a Heading-delayed Mutant dth9 in Rice (Oryza sativa L.)[J]. Chinese Journal OF Rice Science, 2016, 30(3): 232-238.

叶卫军, 胡时开, 吴立文, 郭龙彪, 钱前. 水稻迟抽穗突变体dth9的遗传分析与基因定位[J]. 中国水稻科学, 2016, 30(3): 232-238.

Figures/Tables 8

References 25

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分子标记 Marker	正向引物 (5'-3') Forward primer (5'-3')	反向引物 (5'-3') Reverse primer (5'-3')	大小 Size/bp	物理位置 Physical position/Mb
D9-4	AGCCTCATACCTCCCACA	CGCCTGGAAGACAATCAA	151	3.047
D9-7	AAAGATTCTCAAGGCCAGTC	TATCTAGATCGTGGCCCA	172	3.583
D9-8	TTGCATGGTCACGTTCCT	TGATTGCGGAGTGATGAG	260	3.608
D9-9	CCAATGTAGCAGCCGTAA	CGTTGAGGATTCAGTGGT	129	3.983
D9-17	AATCGGTGAATGTCCTTG	GAAACATCCATGCCTTGC	124	4.223
D9-19	TCCATCGCATTTGAGTGT	AAGTTAGTAGGCGGAAGG	223	4.332
D9-12	GGGGTGATGCTGGTTTAT	AAGGGTCTCATCTGGAAAA	255	4.354
D9-2	GGCTTCTCAACCAAGGTAA	ACGCATCAAATCAGGCAC	205	4.554
RM444	GCTCCACCTGCTTAAGCATC	TGAAGACCATGTTCTGCAGG	162	5.925

分子标记 Marker	正向引物 (5'-3') Forward primer (5'-3')	反向引物 (5'-3') Reverse primer (5'-3')	大小 Size/bp	物理位置 Physical position/Mb
D9-4	AGCCTCATACCTCCCACA	CGCCTGGAAGACAATCAA	151	3.047
D9-7	AAAGATTCTCAAGGCCAGTC	TATCTAGATCGTGGCCCA	172	3.583
D9-8	TTGCATGGTCACGTTCCT	TGATTGCGGAGTGATGAG	260	3.608
D9-9	CCAATGTAGCAGCCGTAA	CGTTGAGGATTCAGTGGT	129	3.983
D9-17	AATCGGTGAATGTCCTTG	GAAACATCCATGCCTTGC	124	4.223
D9-19	TCCATCGCATTTGAGTGT	AAGTTAGTAGGCGGAAGG	223	4.332
D9-12	GGGGTGATGCTGGTTTAT	AAGGGTCTCATCTGGAAAA	255	4.354
D9-2	GGCTTCTCAACCAAGGTAA	ACGCATCAAATCAGGCAC	205	4.554
RM444	GCTCCACCTGCTTAAGCATC	TGAAGACCATGTTCTGCAGG	162	5.925

分子标记 Marker	正向引物 (5'-3') Forward primer (5'-3')	反向引物 (5'-3') Reverse primer (5'-3')
Ghd7	AGGTGCTACGAGAAGCAAATCC	GGGCCTCATCTCGGCATAG
Ghd8	CGTGCAATGGTTTAGACTAAAG	AACAGCATCAGCATCAACAA
Hd6	ACGTGAAGCTATGGCACATC	TGTGGTCGTGCTCTGCTATT
Hd3a	GCTAACGATGATCCCGAT	CCTGCAATGTATAGCATGC
Hd1	CGTTTCGCCAAGAGATCAG	AGATAGAGCTGCAGTGGAGAAC
RFT1	CGTCCATGGTGACCCAACA	CCGGGTCTACCATCACGAGT
Ehd1	AATCGATTCCAACAACAAGCAA	TGTCGAGAGCGGTGGATGA
OsMADS51	GTCGGCAAGCTCTACGAGTACTC	GCGAATTGCTGATAGCGATCA
OsActin1	GCTATGTACGTCGCCATCCA	GGACAGTGTGGCTGACACCAT

分子标记 Marker	正向引物 (5'-3') Forward primer (5'-3')	反向引物 (5'-3') Reverse primer (5'-3')
Ghd7	AGGTGCTACGAGAAGCAAATCC	GGGCCTCATCTCGGCATAG
Ghd8	CGTGCAATGGTTTAGACTAAAG	AACAGCATCAGCATCAACAA
Hd6	ACGTGAAGCTATGGCACATC	TGTGGTCGTGCTCTGCTATT
Hd3a	GCTAACGATGATCCCGAT	CCTGCAATGTATAGCATGC
Hd1	CGTTTCGCCAAGAGATCAG	AGATAGAGCTGCAGTGGAGAAC
RFT1	CGTCCATGGTGACCCAACA	CCGGGTCTACCATCACGAGT
Ehd1	AATCGATTCCAACAACAAGCAA	TGTCGAGAGCGGTGGATGA
OsMADS51	GTCGGCAAGCTCTACGAGTACTC	GCGAATTGCTGATAGCGATCA
OsActin1	GCTATGTACGTCGCCATCCA	GGACAGTGTGGCTGACACCAT

农艺性状 Agronomic trait	野生型 Wild-type	突变体 Mutant
株高Plant height /cm	118.67±1.52	115.30±0.58^*
穗长Panicle length/cm	23.05±0.91	21.33±1.15
有效穗数 No. of effective panicles	8.3±0.5	8.0±0.8
抽穗期 Heading date/d	98.0±0.8	147.3±2.5^**
每穗实粒数 No. of filled grains per panicle	171.5±4.0	173.0±2.7
结实率 Seed-setting rate/%	93.72±0.01	92.20±0.01
千粒重 1000-grain weight/g	31.50±0.57	31.57±0.40
一次枝梗数 Primary rachis branch number	12.0±0.7	12.6±0.6
二次枝梗数 Secondary rachis branch number	36.8±1.3	37.3±0.6