水稻耐金属离子胁迫的QTL分析

doi:10.16819/j.1001-7216.2018.7075

中国水稻科学 ›› 2018, Vol. 32 ›› Issue (1): 23-34.DOI: 10.16819/j.1001-7216.2018.7075

水稻耐金属离子胁迫的QTL分析

林晗¹, 徐江民¹, 胡瑚倩¹, 郑安¹, 徐婉璐¹, 漏平¹, 王跃星², 曾大力^2,^*(), 饶玉春^1,^2,^*()

¹浙江师范大学化学与生命科学学院, 浙江金华 321004
²中国水稻研究所水稻生物学国家重点实验室, 杭州 310006

收稿日期:2017-06-20 修回日期:2017-08-21 出版日期:2018-01-10 发布日期:2018-01-10
通讯作者: 曾大力,饶玉春
作者简介:
共同第一作者:林晗, 徐江民, 胡瑚倩；
基金资助:
国家转基因生物新品种培育重大科技专项(2016ZX08009003-003-008);浙江省自然科学基金资助项目(LY16C130001);浙江省科协育才工程资助项目(2017YCGC008);水稻生物学国家重点实验室开放资助项目

Identifying of QTLs for Resistance to Metal Irons Stress in Rice

Han LIN¹, Jiangmin XU¹, Huqian HU¹, An ZHENG¹, Wanlu XU¹, Ping LOU¹, Yuexing WANG², Dali ZENG^2,^*(), Yuchun RAO^1,^2,^*()

¹College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China
²China National Rice Research Institute, State Key Laboratory of Rice Biology, Hangzhou 310006, China

Received:2017-06-20 Revised:2017-08-21 Online:2018-01-10 Published:2018-01-10
Contact: Dali ZENG, Yuchun RAO
About author:
These authors contributed equally to this work:LIN Han, XU Jiangmin, HU Huqian;

摘要/Abstract

摘要：

【目的】本研究旨在筛选与水稻苗期耐不同金属离子连锁的分子遗传标记,为探讨水稻耐不同金属离子胁迫的遗传研究提供参考。【方法】以典型籼粳交(春江06/台中本地1号)双单倍体(DH)群体为材料,系统考查该群体及其双亲耐4种金属离子(Fe²⁺、Cd²⁺、Al³⁺、Na⁺)胁迫的情况,利用业已构建并完善的该群体加密的分子连锁图谱,对耐这4种金属离子胁迫的QTL进行检测分析。另外,利用实时定量PCR技术检测处理前后相关基因的表达变化情况。【结果】发现耐各种金属离子胁迫的QTL共8个,分别位于水稻第1、2、4、6、9、10和11染色体上,其中Fe²⁺处理后检测到的QTL贡献率最大,达到24.47%(阈值为7.78),位于第1染色体上RM1297–RM1061,同时对该区间与耐胁迫相关基因的表达分析发现这些基因在处理前和处理后表达水平存在不同程度的差异;Cd²⁺处理后检测到1个QTL,位于第1染色体上;Al³⁺处理后检测到QTL共5个,分别位于第2、4、6、10、11染色体上;Na⁺处理后检测到QTL有1个,位于第9染色体上。【结论】根据不同金属离子胁迫处理后DH群体的表型差异进行QTL分析,发现耐各种金属离子胁迫的QTL共8个,并初步定位于各染色体的遗传标记区间,这为精细定位并克隆相应QTL,进而探明水稻耐金属离子胁迫QTL的分子调控机制奠定了基础。

关键词: 水稻, 抗性, 金属胁迫, QTL分析

Abstract:

【Objective】 The molecular markers linked to the tolerance of various metal ions during rice seeding stage were screened for the purpose of discussing the genetic basis of the resistance to different metal ions.【Method】 A rice double haploid(DH) population, derived from a typical indica-japonica cross(Chunjiang 06/TN1) via the anther culture was used to analyze the resistance of four metal ions(Fe²⁺, Cd²⁺, Al³⁺, Na⁺) in DH population and its parents, then the quantitative trait loci (QTL) for the resistance of the four metal ions were identified using an available and complete molecular linkage map. In addition, real-time quantitative PCR was used to detect the expression of related genes before and after treatment.【Result】 A total of 8 QTLs for resistance to metal ions stress were detected, which were localized on chromosomes 1, 2, 4, 6, 9, 10 and 11. Among them, one QTL with the greatest contribution for resistance to Fe²⁺ treatment(24.47%, threshold is 7.78) located in the region RM1297–RM1061 on chromosome 1. Meanwhile, the expression analysis of the genes related to stress tolerance in this interval showed that these genes had different expression levels before and after treatment. One QTL was detected after Cd²⁺ treatment, which was located on chromosome 1. Five QTLs were detected after Al³⁺ treatment, which were located on chromosomes 2, 4, 6, 10 and 11 respectively. And one QTL was detected after Na⁺ treatment, which was located on chromosome 9. 【Conclusion】 According to the QTL analysis of phenotypic differences among DH groups which were treated with different metal ions, eight QTLs for resistance to various metal ions were detected. It laid the foundation for the fine mapping and cloning of the corresponding QTL and exploring the QTL molecular regulation mechanism of the resistance to different metal ions in rice.

Key words: rice, resistance, metal ion stress, QTL analysis

中图分类号:

林晗, 徐江民, 胡瑚倩, 郑安, 徐婉璐, 漏平, 王跃星, 曾大力, 饶玉春. 水稻耐金属离子胁迫的QTL分析[J]. 中国水稻科学, 2018, 32(1): 23-34.

Han LIN, Jiangmin XU, Huqian HU, An ZHENG, Wanlu XU, Ping LOU, Yuexing WANG, Dali ZENG, Yuchun RAO. Identifying of QTLs for Resistance to Metal Irons Stress in Rice[J]. Chinese Journal OF Rice Science, 2018, 32(1): 23-34.

图/表 7

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分子标记 Marker	正向引物(5′-3′) Forward prime(5′-3′)	反向引物(5′-3′) Reverse prime(5′-3′)
OsActin1	TGGCATCTCTCAGCACATTCC	TGCACAATGGATGGGTCAGA
LOC_Os01g50030	AGTGAAAACAGCTGGGCATG	GCATTCTTGTGGTGACCGTT
LOC_Os01g50060	TGTGGGGTTAGCTCTTGGAG	TTTCACCGTTCAGCACCTTG
LOC_Os01g50110	TTTGGCGATCAGTTCTGTGC	TGCTCGTCGTTGATCAGAGA
LOC_Os01g50360	GGCTCTGAATGGGTTTTGGG	ATGTAAGGCAGTCAGGACCC
LOC_Os01g50720	GTGAACAATGCGCTCGAGAG	AAATGTGTCATGCCCTGCTC
LOC_Os01g50820	ATGCTCATCTACTTCCCGCA	CACTCCCGGCTGTAGTACTC
LOC_Os01g50940	CCGCCGGAAGATTTCATGAG	GCCTTGACCACCTCCTTCTT
LOC_Os01g51260	GCTCACGGTGTCACTCAATC	AGAAAGATTGGCTGGTGGGA
LOC_Os01g51430	CTGGGCATGAGAAGTTTCGG	CCAAACAAGAGCAGCCACAT
LOC_Os01g52110	CTTGCGAGAAATGGAGGAGC	CTTGCACTTCTGGCACTTGT
LOC_Os01g52160	TCGCTTTCCATCTAGAGCCA	CATACGGCGTGATATCCCCT
LOC_Os01g52450	TGCTGCTGGGATATATGGCA	CTCCATCCACAGCAATCACG

分子标记 Marker	正向引物(5′-3′) Forward prime(5′-3′)	反向引物(5′-3′) Reverse prime(5′-3′)
OsActin1	TGGCATCTCTCAGCACATTCC	TGCACAATGGATGGGTCAGA
LOC_Os01g50030	AGTGAAAACAGCTGGGCATG	GCATTCTTGTGGTGACCGTT
LOC_Os01g50060	TGTGGGGTTAGCTCTTGGAG	TTTCACCGTTCAGCACCTTG
LOC_Os01g50110	TTTGGCGATCAGTTCTGTGC	TGCTCGTCGTTGATCAGAGA
LOC_Os01g50360	GGCTCTGAATGGGTTTTGGG	ATGTAAGGCAGTCAGGACCC
LOC_Os01g50720	GTGAACAATGCGCTCGAGAG	AAATGTGTCATGCCCTGCTC
LOC_Os01g50820	ATGCTCATCTACTTCCCGCA	CACTCCCGGCTGTAGTACTC
LOC_Os01g50940	CCGCCGGAAGATTTCATGAG	GCCTTGACCACCTCCTTCTT
LOC_Os01g51260	GCTCACGGTGTCACTCAATC	AGAAAGATTGGCTGGTGGGA
LOC_Os01g51430	CTGGGCATGAGAAGTTTCGG	CCAAACAAGAGCAGCCACAT
LOC_Os01g52110	CTTGCGAGAAATGGAGGAGC	CTTGCACTTCTGGCACTTGT
LOC_Os01g52160	TCGCTTTCCATCTAGAGCCA	CATACGGCGTGATATCCCCT
LOC_Os01g52450	TGCTGCTGGGATATATGGCA	CTCCATCCACAGCAATCACG

性状 Trait	阈值 Threshold value	区间 Interval		染色体 Chr.	贡献率 Phenotypic variations explained/%
亚铁胁迫2 d后平均最长根长	7.78	RM1297	–RM1061	1	24.47
The average longest root length after 2 d of ferrous treatment
亚铁胁迫4 d后平均最长根长	7.78	RM1297	–RM1061	1	23.01
The average longest root length after 4 d of ferrous treatment
镉胁迫2 d后平均最长根长	7.96	RM3411	–RM212	1	6.40
The average longest root length after 2 d of cadmium treatment
铝胁迫根平均伸长量	7.69	SSIII-1	–RM3306	4	11.11
The average root elongation after aluminum treatment		RM1370	–SBE1	6	9.29
铝胁迫根相对伸长率	7.69	RM324	–RM341	2	11.66
Relative root elongation rate after aluminum treatment		RM1083	–SSII-1	10	8.68
		RM286	–RM1812	11	8.84
钠胁迫叶片枯死率Withered leaf rate after sodium treatment	7.71	RM1026	–RM205	9	12.21

性状 Trait	阈值 Threshold value	区间 Interval		染色体 Chr.	贡献率 Phenotypic variations explained/%
亚铁胁迫2 d后平均最长根长	7.78	RM1297	–RM1061	1	24.47
The average longest root length after 2 d of ferrous treatment
亚铁胁迫4 d后平均最长根长	7.78	RM1297	–RM1061	1	23.01
The average longest root length after 4 d of ferrous treatment
镉胁迫2 d后平均最长根长	7.96	RM3411	–RM212	1	6.40
The average longest root length after 2 d of cadmium treatment
铝胁迫根平均伸长量	7.69	SSIII-1	–RM3306	4	11.11
The average root elongation after aluminum treatment		RM1370	–SBE1	6	9.29
铝胁迫根相对伸长率	7.69	RM324	–RM341	2	11.66
Relative root elongation rate after aluminum treatment		RM1083	–SSII-1	10	8.68
		RM286	–RM1812	11	8.84
钠胁迫叶片枯死率Withered leaf rate after sodium treatment	7.71	RM1026	–RM205	9	12.21

基因ID Gene ID	基因 Gene	功能 Function
甲基转移酶 Methyltransferase
29610	LOC_Os01g50480	SAM依赖性羧基甲基转移酶 SAM dependent carboxyl methyltransferase
29597	LOC_Os01g50610	SAM依赖性羧基甲基转移酶 SAM dependent carboxyl methyltransferase
50898	LOC_Os01g50030	磷酸乙醇胺N-甲基转移酶 Phosphoethanolamine N-methyltransferase
结构域蛋白 Domain containing protein
50218	LOC_Os01g50940	螺旋-环-螺旋DNA结合结构域蛋白 Helix-loop-helix DNA-binding domain containing protein
50214	LOC_Os01g51140	螺旋-环-螺旋DNA结合结构域蛋白 Helix-loop-helix DNA-binding domain containing protein
29628	LOC_Os01g50360	NAC结构域蛋白 NAC domain containing protein
50236	LOC_Os01g50900	Cupin结构域蛋白 Cupin domain containing protein
50239	LOC_Os01g51010	未知功能结构域292蛋白 DUF292 domain containing protein
49652	LOC_Os01g51990	AN1锌指结构域蛋白 AN1-like zinc finger domain containing protein
16764	LOC_Os01g52410	MYB类DNA结合结构域蛋白 MYB-like DNA-binding domain containing protein
49619	LOC_Os01g52160	重金属相关蛋白结构域 Heavy metal-associated domain containing protein
转录因子 Transcription factor
29592	LOC_Os01g50720	MYB转录因子家族 MYB family transcription factor
50207	LOC_Os01g51260	MYB转录因子家族 MYB family transcription factor
49627	LOC_Os01g52090	转录因子样蛋白 Transcription factor like protein
1822	LOC_Os01g51610	转录因子 Transcription factor
1976	LOC_Os01g50110	MYB转录因子 MYB transcription factor
1805	LOC_Os01g51690	WRKY转录因子 WRKY transcription factor
转运蛋白 Transporter
49616	LOC_Os01g52130	硫酸盐转运蛋白 Sulfate transporter
29598	LOC_Os01g50616	磷脂酰肌醇转移蛋白 Phosphatidylinositol transfer
29637	LOC_Os01g50060	ACC脱氨酶 1-aminocyclopropane-1-carboxylate deaminase
29639	LOC_Os01g50080	MDR类ABC运输蛋白 MDR-like ABC transporter
29641	LOC_Os01g50100	ABC运输蛋白,ATP结合蛋白 ABC transporter, ATP-binding protein
2459	LOC_Os01g50820	高亲和性硝酸盐转运蛋白 High affinity nitrate transporters
蛋白家族 Family protein
29590	LOC_Os01g50700	脱水蛋白家族 Dehydrin family protein
28943	LOC_Os01g51400	富亮氨酸重复蛋白家族 Leucine rich repeat family protein
锌指蛋白 Zinc finger protein
28911	LOC_Os01g51710	锌指结构域蛋白 Zinc knuckle domain containing protein
29595	LOC_Os01g50750	C3HC4型锌指结构域蛋白质 Zinc finger, C3HC4 type domain containing protein
49623	LOC_Os01g52030	锌指蛋白A20和AN1域包含胁迫相关蛋白Zinc finger A20 and AN1 domain-containing stress-associated protein
49639	LOC_Os01g52110	环锌指蛋白 Ring zinc finger protein
酶 Enzyme
16778	LOC_Os01g52450	己糖激酶 Hexokinase
49650	LOC_Os01g51860	紫黄质脱环氧化酶 Violaxanthin de-epoxidase
通道蛋白 Channel protein
49625	LOC_Os01g52070	钾通道AKT1 potassium channel AKT1

水稻耐金属离子胁迫的QTL分析

Identifying of QTLs for Resistance to Metal Irons Stress in Rice

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