茄子萼片覆刺基因的精细定位及分子标记开发

doi:10.7668/hbnxb.20196757

摘要/Abstract

摘要：

为研究茄子萼片覆刺形成的分子机制,以萼片多刺纯合茄子材料24C01为母本,无刺材料24C02为父本,配制遗传群体,F₂萼片覆刺分离呈正态分布。从F₂中分别挑选30株多刺和30株无刺单株,构建刺性状极端混池,利用BSA-seq结合图位克隆技术,将萼片刺基因SmPL定位在6号染色体上131.4 kb区间内。对该区间12个基因进行表达分析,其中仅Smechr0602815.1、Smechr0602817.1和Smechr0602821.1在两亲本间存在差异表达。重测序分析发现,Smechr0602815.1在24C02上存在非同义突变,位于编码区第1 066 bp(G-A),造成谷氨酸突变成赖氨酸;Smechr0602821.1在24C02上存在片段缺失。基于上述序列差异,开发出与无刺基因连锁的KASP标记,对100份不同类型茄子材料检测的符合率均达96%以上,可用于无刺茄子性状基因型的快速鉴定,加快无刺茄子育种进程。

关键词: 茄子, 萼片覆刺, BSA-seq, 精细定位, 分子标记

Abstract:

To elucidate the molecular mechanism of prickle formation on the eggplant calyx,a genetic population was developed by crossing the homozygous prickly line 24C01(female parent)and the non-prickly line 24C02(male parent).In the F₂ generation,the distribution of calyx prickle density followed a normal distribution.Two extreme bulks were constructed by pooling DNA from 30 highly prickly and 30 completely non-prickly F₂ individuals.By combining BSA-seq with map-based cloning,the calyx prickle gene SmPL was fine-mapped to a 131.4 kb interval on chromosome 6.Among the 12 annotated genes within this region,only Smechr0602815.1,Smechr0602817.1,and Smechr0602821.1 showed differential expression between the two parental lines.Re-sequencing analysis further revealed a nonsynonymous mutation(G1066A)in Smechr0602815.1 in 24C02,resulting in a glutamate-to-lysine substitution,and a fragment deletion in Smechr0602821.1 in the same parent.Based on these sequence variations,kompetitive allele-specific PCR(KASP)markers linked to the non-prickly allele were developed.It was found that the markers exhibited a genotype-phenotype concordance rate exceeding 96% when tested across 100 diverse eggplant accessions.These results indicate that the developed KASP markers enable efficient early-selection for the non-prickly trait and can significantly accelerate the breeding of nonprickly eggplant varieties.

Key words: Eggplant, Calyx prickles, BSA-seq, Fine mapping, Molecular marker

中图分类号:

张新改, 刘婧, 乔军, 王利英. 茄子萼片覆刺基因的精细定位及分子标记开发[J]. 华北农学报, 2026, 41(4): 63-74. doi: 10.7668/hbnxb.20196757.

ZHANG Xingai, LIU Jing, QIAO Jun, WANG Liying. Fine Mapping of the Prickle Gene on Eggplant Calyx and Development of Molecular Markers[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(4): 63-74. doi: 10.7668/hbnxb.20196757.

http://www.hbnxb.net/CN/Y2026/V41/I4/63

图/表 14

参考文献 35

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引物名称 Primer	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reverse primer(5'—3')	用途 Function
S2	AGCAAGATTGAGAATTTCGGT	TGGTGAATTAAGTTGTGTTTGG	定位
S33	GCAACTGGTGAGCGAGATTA	TAAGTCCACGCAAGCAAATC
S37	CAGGGATACAGAGGAGAAACC	AAATGATGCGAGAATCAAGC
S43	TAGATAACTGTGCCTCTGATGGA	AACGACTAAAGATCAGATATAGGC
S51	CACATTTGAGAGAAAGGATGC	GTGTAAGCTAGTCGTTGACGG
S67	AAGAAACCAAAATACAGTAACAGTG	TTTTTCATATCTAGTATCTTTGCTCA
K8	1:CAATTTGTTGGAAAATCATACGCGA	ATTTAACTCAACCCGCCTAATTGC
	2:CAATTTGTTGGAAAATCATACGCGG
K9	1:GCGTTTGGAATCTCAGATATGCTG	AGTTATGGGGGAAGAAAGAGCAAT
	2:GCGTTTGGAATCTCAGATATGCTA
SmAPRT	GAGATGCATGTAGGTGCTGTGCAA	GGCCCTTCAATTCTGGCAACTCAA	表达验证
qSm02815	AACAGGTGGGGTTGAAAGGA	ACGAGCTTATCAATTCCTGCCT
qSm02817	CGAGGTGGCTAAGAAGGTAA	CGAGGTGCTAATGACAAGGATA
qSm02821	ACCTGGTGGCTATGGAACGCT	TTTGTGGGGCGTTGGGGCTG
K1	1:AACCTTGACCACCCAAACCTCTTT	GGTAGATTCCAACGAGCAACATGC	KASP标记开发
	2:AACCTTGACCACCCAAACCTCTTC
K2	1:GGCAGCTGATTTGTGTATGCTTGTTG	TCATCCTTCGAGACATCATCAGCAAG
	2:GGCAGCTGATTTGTGTATGCTTGTTA

引物名称 Primer	正向引物(5'—3') Forward primer(5'—3')	反向引物(5'—3') Reverse primer(5'—3')	用途 Function
S2	AGCAAGATTGAGAATTTCGGT	TGGTGAATTAAGTTGTGTTTGG	定位
S33	GCAACTGGTGAGCGAGATTA	TAAGTCCACGCAAGCAAATC
S37	CAGGGATACAGAGGAGAAACC	AAATGATGCGAGAATCAAGC
S43	TAGATAACTGTGCCTCTGATGGA	AACGACTAAAGATCAGATATAGGC
S51	CACATTTGAGAGAAAGGATGC	GTGTAAGCTAGTCGTTGACGG
S67	AAGAAACCAAAATACAGTAACAGTG	TTTTTCATATCTAGTATCTTTGCTCA
K8	1:CAATTTGTTGGAAAATCATACGCGA	ATTTAACTCAACCCGCCTAATTGC
	2:CAATTTGTTGGAAAATCATACGCGG
K9	1:GCGTTTGGAATCTCAGATATGCTG	AGTTATGGGGGAAGAAAGAGCAAT
	2:GCGTTTGGAATCTCAGATATGCTA
SmAPRT	GAGATGCATGTAGGTGCTGTGCAA	GGCCCTTCAATTCTGGCAACTCAA	表达验证
qSm02815	AACAGGTGGGGTTGAAAGGA	ACGAGCTTATCAATTCCTGCCT
qSm02817	CGAGGTGGCTAAGAAGGTAA	CGAGGTGCTAATGACAAGGATA
qSm02821	ACCTGGTGGCTATGGAACGCT	TTTGTGGGGCGTTGGGGCTG
K1	1:AACCTTGACCACCCAAACCTCTTT	GGTAGATTCCAACGAGCAACATGC	KASP标记开发
	2:AACCTTGACCACCCAAACCTCTTC
K2	1:GGCAGCTGATTTGTGTATGCTTGTTG	TCATCCTTCGAGACATCATCAGCAAG
	2:GGCAGCTGATTTGTGTATGCTTGTTA

样本 Sample	比对数据量/Gb Mapped reads	总reads条数 Total reads	比对率/ % Mapping rate	平均深度/× Average depth	至少有1个碱基覆盖的位点占基因组的百分比/% Coverage at least 1×	至少有5个碱基覆盖的位点占基因组的百分比/% Coverage at least 5×
多刺亲本 Prickly parents	9.58	64 553 566	99.81	8.33	89.48	72.74
无刺亲本 Non-prickly parents	11.09	74 684 598	99.87	9.63	89.29	78.12
多刺极端池 Prickly extreme pool	35.03	235 078 230	99.81	30.44	90.96	89.09
无刺极端池 Non-prickly extreme pool	33.96	227 811 532	99.86	29.50	90.94	88.98

样本 Sample	比对数据量/Gb Mapped reads	总reads条数 Total reads	比对率/ % Mapping rate	平均深度/× Average depth	至少有1个碱基覆盖的位点占基因组的百分比/% Coverage at least 1×	至少有5个碱基覆盖的位点占基因组的百分比/% Coverage at least 5×
多刺亲本 Prickly parents	9.58	64 553 566	99.81	8.33	89.48	72.74
无刺亲本 Non-prickly parents	11.09	74 684 598	99.87	9.63	89.29	78.12
多刺极端池 Prickly extreme pool	35.03	235 078 230	99.81	30.44	90.96	89.09
无刺极端池 Non-prickly extreme pool	33.96	227 811 532	99.86	29.50	90.94	88.98

编号 No.	基因名称 Gene name	预测基因功能 Putative function
1	Smechr0602812.1	高亲和硝酸盐转运蛋白2.4样蛋白
2	Smechr0602813.1	高亲和硝酸盐转运蛋白2.4样蛋白
3	Smechr0602814.1	预测蛋白:(R)-特异性烯酰辅酶A水合酶MaoC
4	Smechr0602815.1	类叶绿体Rubisco大亚基结合蛋白α亚基(TCP-1/cpn60家族)
5	Smechr0602816.1	ABC转运蛋白G家族成员28类似物
6	Smechr0602817.1	预测蛋白:LysM结构域受体样激酶 3
7	Smechr0602818.1	功能未知蛋白 EJD97_001307
8	Smechr0602819.1	预测蛋白:类Nectarin-3双功能单脱氢抗坏血酸还原酶与碳酸酐酶
9	Smechr0602820.1	预测蛋白:叶绿体型Nudix水解酶19
10	Smechr0602821.1	预测蛋白:细胞分裂素核苷酸磷酸核糖水解酶LOG4样蛋白
11	Smechr0602822.1	预测蛋白:功能未知蛋白LOC102579416
12	Smechr0602823.1	功能未知蛋白T459_18131

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