水稻OsGMS2基因的鉴定及其核不育系种子繁殖体系构建

doi:10.3724/SP.J.1006.2023.22051

Abstract

Abstract:

Male sterility is the basis for application of crop heterosis. The common nuclear male sterile mutant osgms2 was isolated from a mutant library created by ⁶⁰Co-γ-treated indica cultivar 93-11. The male of the mutant was completely aborted, the female was nomal, and other agronomic characters were consistent with those of the wild type. Genetic analyses indicated that the male sterility phenotype was stably controlled by a single recessive gene. The OsGMS2 gene was fine-mapped within the 86 kb physical interval between two molecular markers S3b and 4826 on the long arm of chromosome 4 with a mapping population of osgms2 and Minghui 63. Further sequencing found the gene LOC_Os04g48490, encoding a fasciclin-like arabinogalactan protein, had a 9-bp deletion at position 118 to 126 from start codon of translation, resulting in three codons mutation. The osgms2 gene was a novel allele of OsFLA1. Subsequent gene knockout and genetic complementation experiments in Zhonghua 11 background confirmed the function of the gene. Real-time quantitative PCR (qPCR) analyses showed that the gene was expressed in all tissues, with the highest level at the flowering and immature seed stages. Alignment analyses revealed that the protein sequence was conserved in different species. By creating the OsGMS2-T maintainer, seed production of the common sterile line was achieved. The identification of the allelic mutant osgms2 provides the new materials for the study on rice nuclear male sterility and application of heterosis.

Key words: rice, genic male sterile, osgms2, gene mapping, OsGMS2-T maintainer line

TANG Jie, LONG Tuan, WU Chun-Yu, LI Xin-Peng, ZENG Xiang, WU Yong-Zhong, HUANG Pei-Jin. Identification of OsGMS2 and construction of seed production system for genic male sterile line in rice[J].Acta Agronomica Sinica, 2023, 49(8): 2025-2038.

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引物名称 Primer name	上游引物 Forward primer (5°-3°)	下游引物 Reverse primer (5°-3°)
RM17332	CGGTACATCACGGTATCAAATCG	TAAATGCTGGAGCGATGCTAACC
RM280	GTGCTCTCCATGTCGGATTATGC	CAAGGCAACAAGATTGGTTAGTGG
RM17351	ATAAAGGAGGAGGGCCTCAGATGG	CACGGTTTGGAGGTTGGAAGC
RM17352	GCTTGGCATCTGCTTCTGTTGTTGG	CTCGCTGCTGATCGAGGTGTCG
RM303	ATCGATGTAGGTAGAGGGACACC	CAGATCTAGTCGACATGGTTGG
4826	ACACCATCTCTCTTCTTTTTCTAT	ATATGGGTAGGTTTGGATATTCG
S4b	GTGTGTGTGAGTAAAATCCTAGTGCA AAAAAGTGTGTGTGAGTAAAATCCTAGAGCC	ATTTGTACTCCTATGTTTAGAATAGC
S3b	ACAAATATATAGCAAAATCGGTGACC AAAAAACAAATATATAGCAAAATCGGTTACG	GTGGTTTTGTGGATGTTTTGTAACT
S2	AAGTATTTGTAATGCACTATGTAAAGGT AAAAAAAGTATTTGTAATGCACTATGTAATGGC	TTAAGAGCACACACTTCCAATAATATGT
S1	CTGGGCGCGGTGCGGCGGGCGAGGC AAAAACTGGGCGCGGTGCGGCGGGCGTGGT	CCGCCTCAGCGCCACCGCCAAGCTGA
S8	AAGTTGTGTTTAGCACTATGTTATTACG AAAAAAAGTTGTGTTTAGCACTATGTTATGACA	TTTAGCATAATAACTACTATTCATCATT
S10	GCAGGAGACACTTGGTGCCGCCTCTC AAAAAGCAGGAGACACTTGGTGCCGCCACTT	GCAGATTATTTTCGGTGGGTCCCGTCTC

引物名称 Primer name	上游引物 Forward primer (5°-3°)	下游引物 Reverse primer (5°-3°)
LOC_Os04g48490_1	AAACAGAAAGCCCCAATG	TGCCGCAGTACGCGCCCAAG
LOC_Os04g48490_2	TTGTCCATGCCGGTGTCCAT	GGTCACGGCACAAACTCA
InD48490	GCTCCGGCTGTTGATCT	GCCTGCTCTTCCTCCTG
GAPDH-RT	GAATGGCTTTCCGTGTT	CAAGGTCCTCCTCAACG
T1-sg	GCGGTCGGTGGCGGCCATGG
SP	CCCGACATAGATGCAATAACTTC	GCGCGGTGTCATCTATGTTACT
B1	AAACCCACGCCCAGAAA	GCCAGGAGGAAGAGCAG
3148 HB	CgcgtttcgaaatttTGATTTCTTCATCGCACT	GtcgcgatcgcatgcACAACATGGTGCAACAGTG

杂交组合 Cross combination	异交结实率 Out-crossing seed-setting rate	F₂		χ²_3:1	χ²_0.05
杂交组合 Cross combination	异交结实率 Out-crossing seed-setting rate	可育株Fertile plants	不育株Sterile plants	χ²_3:1	χ²_0.05
osgms2/93-11	71.27%±14.67%	57	13	0.85	3.841
osgms2/ZH11	62.94%±14.34%	80	16	0.57	3.841

序号 Serial number	基因 Gene name	功能注释 Function annotation
ORF1	LOC_Os04g48390	SPX-Major Facility Superfamily (MFS) protein
ORF2	LOC_Os04g48400	HOTHEAD precursor
ORF3	LOC_Os04g48410	Copper chaperone for superoxide dismutase
ORF4	LOC_Os04g48416	OsSub45-putative subtilisin homologue
ORF5	LOC_Os04g48440	The expressed protein
ORF6	LOC_Os04g48450	The expressed protein
ORF7	LOC_Os04g48460	Cytochrome P450
ORF8	LOC_Os04g48470	The expressed protein
ORF9	LOC_Os04g48480	The exostosin family domain containing protein
ORF10	LOC_Os04g48484	The expressed protein
ORF11	LOC_Os04g48490	The fasciclin-like arabinogalactan protein

Identification of OsGMS2 and construction of seed production system for genic male sterile line in rice

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