水稻籽粒镉积累QTL定位及候选基因分析

doi:10.11983/CBB20148

植物学报 ›› 2021, Vol. 56 ›› Issue (1): 25-32.DOI: 10.11983/CBB20148

水稻籽粒镉积累QTL定位及候选基因分析

潘晨阳¹, 叶涵斐¹, 周维永³, 王盛¹, 李梦佳¹, 路梅¹, 李三峰², 朱旭东², 王跃星², 饶玉春¹^,^*(), 戴高兴³^,^*()

¹浙江师范大学化学与生命科学学院, 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
³广西农业科学院, 南宁 530007

收稿日期:2020-08-26 接受日期:2020-11-11 出版日期:2021-01-01 发布日期:2021-01-15
通讯作者: 饶玉春,戴高兴
作者简介:2526620@qq.com
*E-mail: ryc@zjnu.cn;
基金资助:
国家自然科学基金(31971921);浙江省自然科学杰出青年基金(LR20C130001);浙江省万人计划青年拔尖人才(ZJWR0108023);国家级大学生创新创业训练计划(202010345067);国家级大学生创新创业训练计划(202010345051)

QTL Mapping of Candidate Genes Involved in Cd Accumulation in Rice Grain

Chenyang Pan¹, Hanfei Ye¹, Weiyong Zhou³, Sheng Wang¹, Mengjia Li¹, Mei Lu¹, Sanfeng Li², Xudong Zhu², Yuexing Wang², Yuchun Rao¹^,^*(), Gaoxing Dai³^,^*()

¹College of Chemistry and Life Sciences, Zhejiang Normal University, Jinhua 321004, China
²State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China
³Guangxi Academy of Agricultural Sciences, Nanning 530007, China

Received:2020-08-26 Accepted:2020-11-11 Online:2021-01-01 Published:2021-01-15
Contact: Yuchun Rao,Gaoxing Dai

摘要/Abstract

摘要：

水稻(Oryza sativa)是全世界重要的经济作物之一, 稻田镉(Cd)污染和镉积累问题严重威胁世界水稻的产量和品质以及人类健康, 如何降低水稻中镉积累已成为热点问题。以籼稻品种华占(HZ)为父本、粳稻品种热研2号(Nekken2)为母本, 连续自交多代后得到120个重组自交系群体, 对其镉积累进行检测和分析, 同时利用遗传图谱进行QTL作图。结果共检测到7个QTLs, 分别位于水稻第2、3、9和12号染色体上, 其中1个LOD值高达4.97。对这些QTL区间内与耐金属离子胁迫相关的候选基因进行定量分析, 发现LOC_Os02g50240、LOC_Os02g52780、LOC_Os09g31200、LOC_Os09g35030和LOC_Os09g37949这5个基因在双亲间的表达量差异显著, 结合亲本对不同金属离子的浓度积累数据, 推测LOC_ Os02g50240、LOC_Os09g31200及LOC_Os09g35030的高表达可能极大地提高了水稻对镉离子的吸收和胁迫耐受能力。通过QTL挖掘和分析, 发现这些基因与水稻籽粒的镉积累有关, 可能影响水稻耐镉胁迫的能力。研究结果为进一步筛选和培育耐镉胁迫的水稻品种创造了条件, 为阐明水稻镉积累的分子调控机制奠定了基础。

关键词: 水稻, 镉积累, QTL定位, 候选基因, 离子胁迫, 遗传图谱

Abstract:

Rice is one of the world’s most important crops, but cadmium (Cd) pollution and Cd accumulation in rice fields pose a serious threat to its yield and quality as well as to human health. How to reduce Cd accumulation in rice has become a major concern. In this study, a japonica rice variety Nekken2 was used as the maternal parent, and an indica rice variety HZ was used as the paternal parent. After F₁ generation was obtained by cross-breeding, successive selfings were performed for 120 recombinant inbred lines (RILs) population as the QTL mapping populations. We measured the Cd content in rice grains of each line at the maturity stage. Meanwhile, QTL mapping was performed using genetic map. As a result, a total of 7 QTLs were detected, which were located on chromosome 2, 3, 9, and 12 of rice, with the highest LOD score of 4.97. The qRT-PCR analysis on the candidate genes involved in metal ion stress resistance in these QTL regions showed that the expression level of LOC_Os02g50240, LOC_Os02g52780, LOC_Os09g31200, LOC_Os09g35030 and LOC_Os09g37949 was significantly different from the parents. Combined with the data of concentration of different metal ions in the parents, we speculated that the high expression of LOC_Os02g50240, LOC_Os09g31200 and LOC_Os09g35030 might greatly improve the absorption and tolerance of rice to Cd ion stress. And these genes may be related to the accumulation of Cd in rice grains, and may affect the ability of rice to tolerate Cd stress, which could help for further breeding and screening for rice varieties to tolerate Cd stress, and exploring the molecular regulation mechanism of Cd accumulation in rice.

Key words: rice, cadmium accumulation, QTL mapping, candidate genes, ion stress, genetic map

潘晨阳, 叶涵斐, 周维永, 王盛, 李梦佳, 路梅, 李三峰, 朱旭东, 王跃星, 饶玉春, 戴高兴. 水稻籽粒镉积累QTL定位及候选基因分析. 植物学报, 2021, 56(1): 25-32.

Chenyang Pan, Hanfei Ye, Weiyong Zhou, Sheng Wang, Mengjia Li, Mei Lu, Sanfeng Li, Xudong Zhu, Yuexing Wang, Yuchun Rao, Gaoxing Dai. QTL Mapping of Candidate Genes Involved in Cd Accumulation in Rice Grain. Chinese Bulletin of Botany, 2021, 56(1): 25-32.

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参考文献 30

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Primer name	Sequence (5'-3')	Tm (°C)	Length (bp)
LOC_Os02g50240-F-qrt	CTTCAAGGACCCATTCAGGA	60.04	113
LOC_Os02g50240-R-qrt	TGAAGATCTTGGCAGCATTG	59.95
LOC_Os02g52780-F-qrt	GTTTTCAAAGGTGGGCTGAG	59.71	148
LOC_Os02g52780-R-qrt	CCTCAGCTTCCAATTCCATC	59.68
LOC_Os09g31200-F-qrt	GCGACAAGAAGGATCAGGAG	59.95	117
LOC_Os09g31200-R-qrt	AGAAGCAGTTCTGGCAGAGG	59.75	117
LOC_Os09g35030-F-qrt	GGATCAAGCAGGAGATGAGC	59.92	135
LOC_Os09g35030-R-qrt	GCCTCGTCTCCCTGAACTT	59.39
LOC_Os09g37949-F-qrt	GGAAATCTGCCGATCAGTGT	60.08	123
LOC_Os09g37949-R-qrt	GGGTCCTTCGATGGGTTTAT	60.02

Primer name	Sequence (5'-3')	Tm (°C)	Length (bp)
LOC_Os02g50240-F-qrt	CTTCAAGGACCCATTCAGGA	60.04	113
LOC_Os02g50240-R-qrt	TGAAGATCTTGGCAGCATTG	59.95
LOC_Os02g52780-F-qrt	GTTTTCAAAGGTGGGCTGAG	59.71	148
LOC_Os02g52780-R-qrt	CCTCAGCTTCCAATTCCATC	59.68
LOC_Os09g31200-F-qrt	GCGACAAGAAGGATCAGGAG	59.95	117
LOC_Os09g31200-R-qrt	AGAAGCAGTTCTGGCAGAGG	59.75	117
LOC_Os09g35030-F-qrt	GGATCAAGCAGGAGATGAGC	59.92	135
LOC_Os09g35030-R-qrt	GCCTCGTCTCCCTGAACTT	59.39
LOC_Os09g37949-F-qrt	GGAAATCTGCCGATCAGTGT	60.08	123
LOC_Os09g37949-R-qrt	GGGTCCTTCGATGGGTTTAT	60.02

QTL	Chromosome	Physical distance (bp)	Position of support (cM)	Limit of detection
qRCd-1	2	30302111-35902152	129.90-153.90	4.48
qRCd-2	3	5112031-5335542	21.91-22.87	2.51
qRCd-3	3	6338887-6654517	27.17-28.53	2.61
qRCd-4	9	13883217-15424344	59.51-66.12	4.32
qRCd-5	9	16779765-22230863	71.93-95.30	4.97
qRCd-6	9	22748818-22938953	97.52-98.33	2.60
qRCd-7	12	21976412-22336740	94.21-95.75	2.66

QTL	Chromosome	Physical distance (bp)	Position of support (cM)	Limit of detection
qRCd-1	2	30302111-35902152	129.90-153.90	4.48
qRCd-2	3	5112031-5335542	21.91-22.87	2.51
qRCd-3	3	6338887-6654517	27.17-28.53	2.61
qRCd-4	9	13883217-15424344	59.51-66.12	4.32
qRCd-5	9	16779765-22230863	71.93-95.30	4.97
qRCd-6	9	22748818-22938953	97.52-98.33	2.60
qRCd-7	12	21976412-22336740	94.21-95.75	2.66

Chromosome	Gene	Function	Object
2	LOC_Os02g50240	It participates in the regulation of various biological processes and stress response	Cytoplasmic glutamine synthetase
2	LOC_Os02g52780	It regulates ABA-dependent stress-related gene expression to enhance the plant tolerance of drought and salt	Transcription factors bZIP
9	LOC_Os09g31200	A major determinant of plant response to stress	Zinc finger protein
9	LOC_Os09g35030	It specifically combines with drought response elements and controls many stress-induced genes	Transcription factors AP2/EREBP
9	LOC_Os09g37949	It plays an important role in salt stress response	Receptor protein kinase

水稻籽粒镉积累QTL定位及候选基因分析

QTL Mapping of Candidate Genes Involved in Cd Accumulation in Rice Grain

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