棉花光籽基因n2的精细定位

doi:10.11963/1002-7807.lsmsgl.20190325

摘要/Abstract

摘要：

【目的】棉纤维是由胚珠外珠被表皮单细胞经突起并极性伸长而成的,棉花种子上的短绒是由胚珠外珠被表皮单细胞突起而产生的。因此,本研究对棉纤维光籽基因n₂进行定位,旨在为其克隆和功能研究奠定基础。【方法】以海岛棉新海21和陆地棉光籽突变体n₂为亲本构建F₂群体,结合棉花基因组数据,开发简单序列重复标记,定位该基因,获得预测的基因序列,通过实时荧光定量聚合酶链式反应分析候选基因的表达量差异。【结果】光籽基因n₂被定位在染色体A12上,位于标记P61与Z10之间的181 kbp内。该区间共有7个候选基因。实时荧光定量聚合酶链式反应分析显示,候选基因72098在2个亲本中的表达量有显著差异。【结论】本试验精细定位了光籽基因n₂,初步分析了候选基因,为其图位克隆和功能验证奠定了基础。

关键词: 陆地棉; 纤维; 光籽突变体; 精细定位; 表达分析

Abstract:

[Objective] Cotton fibers are single cells derived from the ovule epidermis, cotton fuzz of seeds are also formed by the ovule epidermis. Therefore, we conduct this experiment to locate the fuzzless gene n₂ so as to lay a foundation for its clone and function study. [Method] Two F₂ populations derived from a cross between the Xinhai 21 and the fuzzless mutant of upland cotton (Gossypium hirsutum L.) n₂ was constructed. Combined with cotton genomic data and developed simple sequence repeat markers to locate the gene. The predicted gene sequence was acquired and the differences in its expression in two parents were analyzed by using the quantitative real time polymerase chain reaction method. [Result] The fuzzless gene was located in a 181 kbp region on chromosome A12 within markers P61 and Z10. There are seven candidate genes in the region. Quantitative real time polymerase chain reaction verification shows significant differences in the expression of 72098 gene in two parents. [Conclusion] In this experiment, the fine mapping and candidate genes analysis of gene n₂ were completed, which laid a foundation for n₂ gene cloning and functional verification.

Key words: Upland cotton; fiber; fuzzless mutant; fine mapping; expression analysis

李思敏,左东云,程海亮,张友平,王巧连,刘珂,冯晓旭,耿洪伟,宋国立. 棉花光籽基因n₂的精细定位[J]. 棉花学报, 2019, 31(2): 114-120.

Li Simin,Zuo Dongyun,Cheng Hailiang,Zhang Youping,Wang Qiaolian,Liu Ke,Feng Xiaoxu,Geng Hongwei,Song Guoli. Fine Mapping of the Fuzzless Gene n₂ in Cotton[J]. Cotton Science, 2019, 31(2): 114-120.

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链接本文: https://journal.cricaas.com.cn/Jweb_mhxb/CN/10.11963/1002-7807.lsmsgl.20190325

https://journal.cricaas.com.cn/Jweb_mhxb/CN/Y2019/V31/I2/114

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参考文献 29

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基因名称 Gene name	正向引物 Forward primer	反向引物 Reverse primer
1414	CTCCCGCACCTTAGCCGAGCGTATC	CTAACAAATTTTGTTCTTGTTAGTT
1415	ATGGGTTTTAGCTTGAAAGCTTACT	GCCTTTGCGTTCCCAGCAGCTCCTA
72098	ATGCATGTGGGAGGCATTGCAGCAG	AACCATGATGGATAGCTGAAAGAAG
1417	ATGGCAAAAGAAAAAGAAAGCCACC	CGGCAATGGTGGGGATAACCTAAAG
1418	ATGGAGAGGGGGTCTTTTTTGCGGA	TACGGGGATCTGATCGACCCATTTT
1419	CGCGATCATCATCATCAAGACCT	TCAGGCAAACAAGAGAAGCC
1420	CGATTCTCGTGCTCTCTCCG	AAAATTCAACACAGCTTCCACAA
ACTIN	ATCCTCCGTCTTGACCTTG	TGTCCGTCAGGCAACTCAT

编号 Number	标记 Marker	正向引物 Forward primer	反向引物 Reverse primer
1	NAU3294	ACGATGTCGAGCTACCTTTC	ATTTGTGATTTCTGCGGATT
2	NAU2176	TGGCACTTGCTAAACCATAA	GGTTTGAGCAGAAGGGTATG
3	NAU1231	TTGAGACCAAAAACATGTGG	GCTCATTTTGATCTGAACTCTG
4	NAU2096	GAGAGATAAACGGAGGTGGA	TTAGTCAAACCGAGCAGCTT
5	NAU6112	TCTGTGAACATCAAGTGCTA	TCATAAACTAATGCTAGACAGG
6	NAU3236	AACATCCCCATCAAACACAT	CCCCATGCTGTCTTGTTACT
7	P80	GCATCATACGAAGACGAACG	TGTCGACTAGTTGTTGATCATGC
8	C28	TTGGAGACCAAAACAGACGGT	CAGCTGAGTCGATGCTCTCT
9	NAU0943	ATCTGTTCAATTTCTCGTCA	CAGTTGTTGGTTGATCTGGA
10	P6	CGTAGAAACCCTTACGGGAA	TCTTTGGTGAAGGTATTGCAT
11	P11	TGACTTAATTTGACTTACCAATTTGA	GAAGCCGTTTATCGTGAGGA
12	P16	GGGAGATGTGGTGAAGTTGG	TTTCATGCTTTCTGACCAGG
13	P32	TCGCTAACTCAAGCCCACTT	GGCTCGAACCAATCCAATTT
14	P61	GGCCTTGGACTTCAATTTGT	CAACTAAACGCGCTTTCACA
15	P77	AAATGCACTAATAATAGCACGGG	TTGATTTCAATTTGATCCTTGG
16	P81	AATAGCATGGCCATAATCACA	TCATTCTAATTTCAATTTGATCCTTG
17	C2	GGGCATTTCAGACCATCCCA	CGGGCTTAGAGAGCCGAAAT
18	Z10	TTAGATCTCTGGTTCAACTGTT	TTGCCTTGCCTTGCCAATCTC

基因名称 Gene name	功能注释 Function annotation	基因全长 Full length of gene/bp
1414	谷氨酸脱羧酶4 Glutamate decarboxylase 4	1 500
1415	双功能抑制剂/脂质转运蛋白Bifunctional inhibitor/lipid-transfer protein	357
72098	蛋白激酶Protein kinase	381
1417	C₂H₂锌指蛋白C₂H₂-like zinc finger protein	1 074
1418	铝活化苹果酸转运蛋白Aluminum-activated malate transporter family protein	1 980
1419	ZIP金属离子转运蛋白ZIP metal ion transporter family	1 410
1420	细胞周期蛋白Cyclin J18	711

棉花光籽基因n₂的精细定位

Fine Mapping of the Fuzzless Gene n₂ in Cotton

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