甘蓝型油菜角果数突变体基因的定位及候选基因分析

doi:10.3724/SP.J.1006.2022.04281

Abstract

Abstract:

The silique number is one of the important components of yield per plant in oilseed rape (Brassica napus L.) and the exploitation and utilization of its excellent alleles are essential to increase yield. More than hundreds of silique number QTLs have been mapped in oilseed rape, but they are difficult to be fine-mapped or cloned because of their moderate and unstable effects. A oilseed rape mutant (No.7931) was detected in previous study and it had few siliques at mature stage due to the stop growth after differentiation about 10 flowers on the top of inflorescence. A F₂ segregating population consisting of 3400 individuals was constructed using this mutant and another more-silique lines No.73290. Among them, we performed BSA-seq on 30 individuals with extreme more- or less-siliques and detected three associated intervals of 0-1.1 Mb, 4.7-6.2 Mb, and 11.5-12.4 Mb on the C02 chromosome. These genomic intervals contained a total of 522 annotated genes in the reference genome DarmorV8.1, among which 235 genes had functional annotation and SNP/InDel variation. At the early stage of flower bud differentiation, the shoot apical meristems of two parents were subjected to RNA-seq, and a total of 8958 differentially expressed genes (DEGs) were detected. These DEGs were significantly enriched into 20 pathways, including carbohydrate metabolism, translation, and amino acid metabolism (highly associated with flower bud differentiation) and so on, among which 99 were located in the associated intervals. By the integration of gene functional annotation as well as sequence and expression variation analysis, a total of nine candidate genes (BnaC02g00490.1D2, BnaC02g01030.1D2, BnaC02g01120.1D2, BnaC02g00270.1D2, BnaC02g02670.1D2, BnaC02g08680.1D2, BnaC02g08890.1D2, BnaC02g09480.1D2, and BnaC02g10490.1D2) were identified, which were mainly involved in the maintenance of inflorescence meristems and the regulation of flower development. The above results lay the foundation for the following fine-mapping and cloning of the silique number mutant gene in oilseed rape.

Key words: Brassica napus L., Brassica napus L., silique number mutant, silique number mutant, flower bud differentiation, flower bud differentiation, BSA-seq, BSA-seq, RNA-seq, RNA-seq

ZHAO Gai-Hui, LI Shu-Yu, ZHAN Jie-Peng, LI Yan-Bin, SHI Jia-Qin, WANG Xin-Fa, WANG Han-Zhong. Mapping and candidate gene analysis of silique number mutant in Brassica napus L.[J].Acta Agronomica Sinica, 2022, 48(1): 27-39.

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亲本 Parent	主花序花器官数目 FNm	主花序角果数 SNm	株高 PH
No.7931	8.33±1.24	7.67±0.94	154.3±2.06
No.73290	199.3±1.24	93.67±2.49	150.4±1.57
t检验值t-test value	5.45E-09	6.91E-07	0.0170

亲本 Parent	主花序花器官数目 FNm	主花序角果数 SNm	株高 PH
No.7931	8.33±1.24	7.67±0.94	154.3±2.06
No.73290	199.3±1.24	93.67±2.49	150.4±1.57
t检验值t-test value	5.45E-09	6.91E-07	0.0170

基因 Gene	正向引物 Forward primer (5°-3°)	反向引物 Reverse primer (5°-3°)
BnaA02G0314300ZS	CCGACACGCTTCAGAAGGTC	AGAGATCCCGCTTCGACTCC
BnaC03G0027000ZS	TGAGAACACGCCGGTCAAAG	CCTCGCGTTCTCTCTCTCCA
BnaC03G0100100ZS	CTGATGGCCGTAGAGCATGC	AGCCATCTCCTGTTGTTGCA
BnaC03G0091000ZS	TCCCACTCTACCCTGCACTG	GACGTTGTTCACATTCGCGC
BnaC02G0283700ZS	ACAACATGGCCCTGGAAACTG	CTTGGAGGCGGATGATCGTT
BnaC02G0139700ZS	CGGCGGAGGTACAGACATTT	TCGTCATGATGAGCCTCTCCT
BnaC02G0175400ZS	GCGCCTCGTATCCATTCTCG	CCTGAAGTTGTGGCGAGCTT
BnaA01G0077800ZS	CCACCGTCATGTCTTCCTTCC	ACGAGTTGGAAGTGTGCGTT
BnaA01G0061200ZS	GGGAACGGCTTAGATGGTGC	TTGCTAGTACCAGGGCTGCT
BnaC02G0013900ZS	CATTGGCTCGTCATGAACATC	TTCACGAGTGTTGAACTGATCC
BnaC02G0159100ZS	CTGTCACTGGAAACCACCCG	TCAATCGACCATGGCAAGCA
BnaA01G0009100ZS	TCTGCTCTGAACGCGACCAA	ACCAGCCAAAGAACCAGGGT
BnaA01G0037200ZS	TCCTCAACTGTGCCGACATGT	AAAGCCGTCGTCAATCTCGC
BnaC05G0266700ZS	AGACTACGTGAAGCAGCCGA	CTCCAGCTTCCGACCAGTCT
BraActin	CTGGAATTGCTGACCGTATGAG	ATCTGTTGGAAAGTGCTGAGGG

基因 Gene	正向引物 Forward primer (5°-3°)	反向引物 Reverse primer (5°-3°)
BnaA02G0314300ZS	CCGACACGCTTCAGAAGGTC	AGAGATCCCGCTTCGACTCC
BnaC03G0027000ZS	TGAGAACACGCCGGTCAAAG	CCTCGCGTTCTCTCTCTCCA
BnaC03G0100100ZS	CTGATGGCCGTAGAGCATGC	AGCCATCTCCTGTTGTTGCA
BnaC03G0091000ZS	TCCCACTCTACCCTGCACTG	GACGTTGTTCACATTCGCGC
BnaC02G0283700ZS	ACAACATGGCCCTGGAAACTG	CTTGGAGGCGGATGATCGTT
BnaC02G0139700ZS	CGGCGGAGGTACAGACATTT	TCGTCATGATGAGCCTCTCCT
BnaC02G0175400ZS	GCGCCTCGTATCCATTCTCG	CCTGAAGTTGTGGCGAGCTT
BnaA01G0077800ZS	CCACCGTCATGTCTTCCTTCC	ACGAGTTGGAAGTGTGCGTT
BnaA01G0061200ZS	GGGAACGGCTTAGATGGTGC	TTGCTAGTACCAGGGCTGCT
BnaC02G0013900ZS	CATTGGCTCGTCATGAACATC	TTCACGAGTGTTGAACTGATCC
BnaC02G0159100ZS	CTGTCACTGGAAACCACCCG	TCAATCGACCATGGCAAGCA
BnaA01G0009100ZS	TCTGCTCTGAACGCGACCAA	ACCAGCCAAAGAACCAGGGT
BnaA01G0037200ZS	TCCTCAACTGTGCCGACATGT	AAAGCCGTCGTCAATCTCGC
BnaC05G0266700ZS	AGACTACGTGAAGCAGCCGA	CTCCAGCTTCCGACCAGTCT
BraActin	CTGGAATTGCTGACCGTATGAG	ATCTGTTGGAAAGTGCTGAGGG

油菜基因编号 B. napus ID	拟南芥基因名 Name of A. thaliana	拟南芥基因编号 A. thaliana ID	位置Position (Mb)	基因注释 Gene annotation	序列差异 Sequence difference	差异表达基因 Differential expressed genes
BnaC02g00490.1D2	FLC	AT5g10140	0.208	K盒区和MADS转录因子家族蛋白 K-box region and MADS-box transcription factor family protein	有Yes	是No
BnaC02g01030.1D2	TPR1	AT1G80490	0.465	TPL相关的基因1 TOPLESS-related 1	无No	否No
BnaC02g01120.1D2	AGAL2	AT5G08370	0.493	α-半乳糖苷酶2 Alpha-galactosidase 2	无No	否No
BnaC02g00270.1D2	AIL6	AT5G10510	0.996	类ANT基因6 AINTEGUMENTA-LIKE 6	有Yes	否No
BnaC02g02670.1D2	TFL1	AT5G03840	1.10	磷脂酰乙醇胺结合蛋白 Phosphatidylethanolamine-binding-protein	无No	是Yes
BnaC02g08680.1D2	COL1	AT5G15850	4.83	类CO基因1 CONSTANS-like 1	有Yes	否No
BnaC02g08890.1D2	ELF9	AT5G16260	4.96	RNA结合(RRM/RBD/RNP基序)家族蛋白RNA binding (RRM/RBD/RNP motifs) family protein	有Yes	否No
BnaC02g09480.1D2	AIM1	AT4G29010	5.23	烯酰辅酶A水合酶/异构酶家族 Enoyl-CoA hydratase/isomerase family	有Yes	否No
BnaC02g10490.1D2	CHR17	AT5G18620	5.97	染色质重塑因子17 Chromatin remodeling factor 17	无No	是Yes

Mapping and candidate gene analysis of silique number mutant in Brassica napus L.

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