水稻抗氧化性状遗传位点挖掘及候选基因分析

doi:10.11983/CBB24065

植物学报 ›› 2024, Vol. 59 ›› Issue (5): 738-751.DOI: 10.11983/CBB24065 cstr: 32102.14.CBB24065

所属专题：大食物观

水稻抗氧化性状遗传位点挖掘及候选基因分析

连锦瑾¹^,^†, 唐璐瑶¹^,^†, 张伊诺¹, 郑佳兴¹, 朱超宇¹, 叶语涵¹, 王跃星², 商文楠³, 傅正浩¹, 徐昕璇¹, 吴日成¹, 路梅¹, 王长春¹^,^*(), 饶玉春¹^,^*()

¹浙江师范大学生命科学学院, 金华 321004
²中国水稻研究所水稻生物育种全国重点实验室, 杭州 310006
³五常市商博士米业有限公司, 哈尔滨 150200

收稿日期:2024-04-30 接受日期:2024-06-21 出版日期:2024-09-10 发布日期:2024-08-19
通讯作者: 王长春,饶玉春
作者简介:第一联系人：† 共同第一作者
基金资助:
浙江省自然科学基金重点项目(LZ23C130003);2024年国家级大学生创新创业训练计划项目和2024年浙江省大学生科技创新活动计划暨新苗人才计划

Genetic Locus Mining and Candidate Gene Analysis of Antioxidant Traits in Rice

Lian Jinjin¹^,^†, Tang Luyao¹^,^†, Zhang Yinuo¹, Zheng Jiaxing¹, Zhu Chaoyu¹, Ye Yuhan¹, Wang Yuexing², Shang Wennan³, Fu Zhenghao¹, Xu Xinxuan¹, Wu Richeng¹, Lu Mei¹, Wang Changchun¹^,^*(), Rao Yuchun¹^,^*()

¹College of Life Sciences, Zhejiang Normal University, Jinhua 321004, China
²State Key Laboratory of Rice Biology, China National Rice Research Institute, Hangzhou 310006, China
³Wuchang Dr. Shang Rice Industry Co., Ltd., Harbin 150200, China

Received:2024-04-30 Accepted:2024-06-21 Online:2024-09-10 Published:2024-08-19
Contact: Wang Changchun, Rao Yuchun
About author:First author contact:† These authors contributed equally to this paper

摘要/Abstract

摘要： 水稻(Oryza sativa)是世界上最重要的粮食作物之一。提高水稻抗氧化性, 进而提高抗逆性是保障其高产稳产的重要途径。选用籼稻华占(HZ)和粳稻热研2号(Nekken2)及以其为亲本构建的120个重组自交系(RILs), 分别在分蘖期、灌浆期和成熟期测定亲本及其后代剑叶、颖壳及籽粒中羟基自由基清除率、总酚含量、黄酮含量和花青苷含量, 同时基于已构建的高密度遗传连锁图谱进行数量性状基因座(QTL)定位。结果共挖掘到62个与水稻抗氧化损伤有关的QTLs, 其中LOD值最高达4.36。对这些QTL区间内相关候选基因的表达进行定量分析, 结果表明LOC_Os06g01850、LOC_Os12g07820、LOC_ Os12g07830和LOC_Os03g60509等13个基因的表达在不同时期的双亲间差异显著。研究挖掘到众多与水稻抗氧化性相关的QTLs, 为进一步定位并克隆相关基因, 选育抗性强且营养价值高的水稻新品种奠定基础。

关键词: 水稻, 抗氧化性, 遗传图谱, QTL定位

Abstract: Rice (Oryza sativa) is one of the most important food crops in the world. Improving its antioxidant ability and stress resistance is an important way to ensure high and stable yields. In this study, we used the indica rice HZ and the japonica rice Nekken2 as parents and the 120 recombinant inbred line population constructed from them as experimental materials to determine the hydroxyl radical scavenging rate, total phenol content, flavonoid content, and anthocyanin content in sword leaves, glume shells and grains of parents and their progeny at three stages: the tillering stage, the grain filling stage and the maturity stage. Additionally, a total of 62 QTLs related to rice antioxidant damage were identified on the basis of the constructed high-density genetic map for QTL mapping, with an LOD value of up to 4.36. A quantitative analysis of candidate genes related to antioxidant damage ability in these regions revealed that thirteen candidate genes, including LOC_Os06g01850, LOC_Os12g07820, LOC_Os12g07830, and LOC_Os03g60509 were significantly differentially expressed between the two parents at different growth stages. A multitude of QTLs associated with antioxidant damage resistance in rice were identified, providing a foundation for further mapping and cloning of related genes and the development of new rice varieties with increased resistance and nutritional value.

Key words: rice, antioxidant ability, genetic map, QTL mapping

连锦瑾, 唐璐瑶, 张伊诺, 郑佳兴, 朱超宇, 叶语涵, 王跃星, 商文楠, 傅正浩, 徐昕璇, 吴日成, 路梅, 王长春, 饶玉春. 水稻抗氧化性状遗传位点挖掘及候选基因分析. 植物学报, 2024, 59(5): 738-751.

Lian Jinjin, Tang Luyao, Zhang Yinuo, Zheng Jiaxing, Zhu Chaoyu, Ye Yuhan, Wang Yuexing, Shang Wennan, Fu Zhenghao, Xu Xinxuan, Wu Richeng, Lu Mei, Wang Changchun, Rao Yuchun. Genetic Locus Mining and Candidate Gene Analysis of Antioxidant Traits in Rice. Chinese Bulletin of Botany, 2024, 59(5): 738-751.

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图/表 7

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Physiological traits	Gene name	Forward primer (5'-3')	Reverse primer (5′-3′)
	OsActin	TGGCATCTCAGCACATTCC	TGCACAATGGATGGGTCAGA
Hydroxyl radical scavenging rate	LOC_Os06g01850	ACATGGTCTTCAGCACCGAG	CTTTGCCTACTGGTCCGGTT
	LOC_Os06g48000	TTCAACCTCGACCAGCTCAC	GGAGGTTCTGGTAGAAGGCG
	LOC_Os06g48020	GTTGTGACGCGTCGATCATG	CCGCTCTGGTAGACGGATTC
Total phenolic content	LOC_Os04g45210	AGCTGGTCAAGACGTTCCAG	GAAGTAGACCATCTCGCCGG
	LOC_Os12g07820	GTTCGGTTAGGGTGGCATGA	CATTGTTCAGGGGCAGCAAC
	LOC_Os12g07830	CCTCTCCCCTCCCTTCTCTC	GTCATGCCACCCTAACCGAA
Flavonoid content	LOC_Os05g12190	GACCTCAAGCTGGCATCACT	CCCAAACAGTCCATGACCGA
	LOC_Os05g12210	ATGCCCTCCCTTGAAACTCG	TCACGGTTGTTCTCGGTGAG
	LOC_Os05g12240	GGCCTCAACCCTTCTGTCTC	AGGACCATCTCAAACAGCGG
	LOC_Os05g45120	CTCATGGTGGAGGACATGGG	CAACAGGAACTCGGCAAACG
	LOC_Os12g16260	TGATACGCAACTTCTACACTGCT	AGATGGCTCAGGTTGGTTGG
Anthocyanin content	LOC_Os03g60509	TGTGTTCCGTGTCAGTGGAG	CAGGATCATCGTCACCCTCG
Anthocyanin content	LOC_Os03g62480	CCATCCCCGCCTACTTCTTC	ATCGAAGCTGTTCACGAGCA

Physiological traits	Gene name	Forward primer (5'-3')	Reverse primer (5′-3′)
	OsActin	TGGCATCTCAGCACATTCC	TGCACAATGGATGGGTCAGA
Hydroxyl radical scavenging rate	LOC_Os06g01850	ACATGGTCTTCAGCACCGAG	CTTTGCCTACTGGTCCGGTT
	LOC_Os06g48000	TTCAACCTCGACCAGCTCAC	GGAGGTTCTGGTAGAAGGCG
	LOC_Os06g48020	GTTGTGACGCGTCGATCATG	CCGCTCTGGTAGACGGATTC
Total phenolic content	LOC_Os04g45210	AGCTGGTCAAGACGTTCCAG	GAAGTAGACCATCTCGCCGG
	LOC_Os12g07820	GTTCGGTTAGGGTGGCATGA	CATTGTTCAGGGGCAGCAAC
	LOC_Os12g07830	CCTCTCCCCTCCCTTCTCTC	GTCATGCCACCCTAACCGAA
Flavonoid content	LOC_Os05g12190	GACCTCAAGCTGGCATCACT	CCCAAACAGTCCATGACCGA
	LOC_Os05g12210	ATGCCCTCCCTTGAAACTCG	TCACGGTTGTTCTCGGTGAG
	LOC_Os05g12240	GGCCTCAACCCTTCTGTCTC	AGGACCATCTCAAACAGCGG
	LOC_Os05g45120	CTCATGGTGGAGGACATGGG	CAACAGGAACTCGGCAAACG
	LOC_Os12g16260	TGATACGCAACTTCTACACTGCT	AGATGGCTCAGGTTGGTTGG
Anthocyanin content	LOC_Os03g60509	TGTGTTCCGTGTCAGTGGAG	CAGGATCATCGTCACCCTCG
Anthocyanin content	LOC_Os03g62480	CCATCCCCGCCTACTTCTTC	ATCGAAGCTGTTCACGAGCA

Physiological traits	QTL	Chromosome	Physical distance (bp)	Position of support (cM)	Limit of detection
Hydroxyl radical scavenging rate	qHRSR1.1	1	2556386-2834929	10.96-12.15	2.14
	qHRSR2.1	2	3467582-4955784	14.87-21.24	4.36
	qHRSR2.2	2	5098574-6084984	21.86-26.08	3.37
	qHRSR2.3	2	27009300-27239796	115.78-116.77	2.75
	qHRSR2.4	2	28427871-28664694	121.86-122.88	2.21
	qHRSR3.1	3	12723031-12925845	54.54-55.41	2.22
	qHRSR3.2	3	29949770-30321190	128.39-129.98	2.31
	qHRSR4.1	4	23316226-23991142	99.95-102.84	3.35
	qHRSR4.2	4	23680403-239113 17	101.51-102.50	2.00
	qHRSR4.3	4	24725621-25022230	105.99-107.26	2.29
	qHRSR5.1	5	26927610-27082669	115.43-116.10	2.30
	qHRSR6.1	6	33441-781344	0.14-3.35	2.50
	qHRSR6.2	6	26490081-27333693	113.56-117.17	2.24
	qHRSR6.3	6	28674720-29130039	123.31-124.87	2.62
	qHRSR8.1	8	1092742-2649021	4.68-11.36	2.97
	qHRSR8.2	8	11086291-11153903	47.52-47.81	2.22
	qHRSR8.3	8	14983464-15034055	64.23-64.45	2.36
	qHRSR8.4	8	21388530-22202508	91.69-95.18	2.44
Total phenolic content	qTPC1.1	1	7667525-8235815	32.87-35.30	2.23
	qTPC3.1	3	15484683-16021103	66.38-68.68	3.07
	qTPC3.2	3	16899768-17721978	72.45-75.97	3.58
	qTPC3.3	3	25032115-25648379	107.31-109.95	2.50
	qTPC4.1	4	26377931-27479589	113.07-117.80	2.44
	qTPC12.1	12	2689107-3043767	11.53-13.05	2.52
	qTPC12.2	12	3933801-4462301	16.86-19.13	2.08
	qTPC12.3	12	11398960-11895586	48.86-50.99	2.00
	qTPC12.4	12	13100229-14071703	56.16-60.32	2.00
	qTPC12.5	12	14215593-15112427	60.94-64.78	2.11
	qTPC12.6	12	16682032-1680484	71.51-72.19	2.47
Flavonoid content	qFC1.1	1	26148806-27178896	112.09-116.50	2.42
	qFC1.2	1	27178896-27631674	116.50-118.45	2.03
	qFC1.3	1	37113012-37638053	159.09-161.34	2.12
	qFC1.4	1	38157798-38395992	163.57-164.59	2.35
	qFC2.1	2	21915215-22231364	93.94-95.30	2.30
	qFC4.1	4	17472715-19028032	74.90-81.57	3.67
	qFC4.2	4	19642144-19986484	84.20-85.68	2.70
	qFC4.3	4	22019563-23235126	94.39-99.60	3.74
	qFC4.4	4	26377931-26619819	113.07-114.11	2.87
	qFC5.1	5	5206830-6439595	22.32-27.60	4.03
	qFC5.2	5	6673100-7059585	28.61-30.26	4.12
	qFC5.3	5	12422042-12794190	53.25-54.85	2.04
	qFC5.4	5	26189817-26927610	112.27-115.43	2.59
	qFC6.1	6	5187517-6015262	22.24-25.79	3.28
	qFC6.2	6	6069227-6560451	26.02-28.12	2.68
	qFC6.3	6	21068005-21507916	90.31-92.20	2.16
	qFC6.4	6	27961504-28384769	119.86-121.68	2.51
	qFC9.1	9	13119644-13620974	56.24-58.39	2.07
	qFC10.1	10	21806620-22080253	93.48-94.65	2.02
	qFC12.1	12	2351235-2689107	10.08-11.53	2.56
	qFC12.2	12	4462301-6224960	19.13-26.68	2.95
	qFC12.3	12	7508660-9990803	32.19-42.83	3.74
	qFC12.4	12	11398960-14215593	48.86-60.94	3.04
	qFC12.5	12	17056555-17574928	73.12-75.34	2.19
	qFC12.6	12	23839134-24088353	102.19-103.26	2.39
Anthocyanin content	qAC2.1	2	35599368-35902152	152.60-153.90	2.42
	qAC3.1	3	31965505-32493480	137.03-138.32	2.25
	qAC3.2	3	33386000-34509821	143.12-147.93	3.23
	qAC3.3	3	35314343-35646346	151.38-152.81	2.30
	qAC5.1	5	22147125-22524107	94.94-96.55	2.88
	qAC11.1	11	9832804-10200412	42.15-43.73	2.28
	qAC12.1	12	927518-2689107	3.98-11.53	2.31
	qAC12.2	12	23013674-23469132	98.65-100.61	2.50

Physiological traits	QTL	Chromosome	Physical distance (bp)	Position of support (cM)	Limit of detection
Hydroxyl radical scavenging rate	qHRSR1.1	1	2556386-2834929	10.96-12.15	2.14
	qHRSR2.1	2	3467582-4955784	14.87-21.24	4.36
	qHRSR2.2	2	5098574-6084984	21.86-26.08	3.37
	qHRSR2.3	2	27009300-27239796	115.78-116.77	2.75
	qHRSR2.4	2	28427871-28664694	121.86-122.88	2.21
	qHRSR3.1	3	12723031-12925845	54.54-55.41	2.22
	qHRSR3.2	3	29949770-30321190	128.39-129.98	2.31
	qHRSR4.1	4	23316226-23991142	99.95-102.84	3.35
	qHRSR4.2	4	23680403-239113 17	101.51-102.50	2.00
	qHRSR4.3	4	24725621-25022230	105.99-107.26	2.29
	qHRSR5.1	5	26927610-27082669	115.43-116.10	2.30
	qHRSR6.1	6	33441-781344	0.14-3.35	2.50
	qHRSR6.2	6	26490081-27333693	113.56-117.17	2.24
	qHRSR6.3	6	28674720-29130039	123.31-124.87	2.62
	qHRSR8.1	8	1092742-2649021	4.68-11.36	2.97
	qHRSR8.2	8	11086291-11153903	47.52-47.81	2.22
	qHRSR8.3	8	14983464-15034055	64.23-64.45	2.36
	qHRSR8.4	8	21388530-22202508	91.69-95.18	2.44
Total phenolic content	qTPC1.1	1	7667525-8235815	32.87-35.30	2.23
	qTPC3.1	3	15484683-16021103	66.38-68.68	3.07
	qTPC3.2	3	16899768-17721978	72.45-75.97	3.58
	qTPC3.3	3	25032115-25648379	107.31-109.95	2.50
	qTPC4.1	4	26377931-27479589	113.07-117.80	2.44
	qTPC12.1	12	2689107-3043767	11.53-13.05	2.52
	qTPC12.2	12	3933801-4462301	16.86-19.13	2.08
	qTPC12.3	12	11398960-11895586	48.86-50.99	2.00
	qTPC12.4	12	13100229-14071703	56.16-60.32	2.00
	qTPC12.5	12	14215593-15112427	60.94-64.78	2.11
	qTPC12.6	12	16682032-1680484	71.51-72.19	2.47
Flavonoid content	qFC1.1	1	26148806-27178896	112.09-116.50	2.42
	qFC1.2	1	27178896-27631674	116.50-118.45	2.03
	qFC1.3	1	37113012-37638053	159.09-161.34	2.12
	qFC1.4	1	38157798-38395992	163.57-164.59	2.35
	qFC2.1	2	21915215-22231364	93.94-95.30	2.30
	qFC4.1	4	17472715-19028032	74.90-81.57	3.67
	qFC4.2	4	19642144-19986484	84.20-85.68	2.70
	qFC4.3	4	22019563-23235126	94.39-99.60	3.74
	qFC4.4	4	26377931-26619819	113.07-114.11	2.87
	qFC5.1	5	5206830-6439595	22.32-27.60	4.03
	qFC5.2	5	6673100-7059585	28.61-30.26	4.12
	qFC5.3	5	12422042-12794190	53.25-54.85	2.04
	qFC5.4	5	26189817-26927610	112.27-115.43	2.59
	qFC6.1	6	5187517-6015262	22.24-25.79	3.28
	qFC6.2	6	6069227-6560451	26.02-28.12	2.68
	qFC6.3	6	21068005-21507916	90.31-92.20	2.16
	qFC6.4	6	27961504-28384769	119.86-121.68	2.51
	qFC9.1	9	13119644-13620974	56.24-58.39	2.07
	qFC10.1	10	21806620-22080253	93.48-94.65	2.02
	qFC12.1	12	2351235-2689107	10.08-11.53	2.56
	qFC12.2	12	4462301-6224960	19.13-26.68	2.95
	qFC12.3	12	7508660-9990803	32.19-42.83	3.74
	qFC12.4	12	11398960-14215593	48.86-60.94	3.04
	qFC12.5	12	17056555-17574928	73.12-75.34	2.19
	qFC12.6	12	23839134-24088353	102.19-103.26	2.39
Anthocyanin content	qAC2.1	2	35599368-35902152	152.60-153.90	2.42
	qAC3.1	3	31965505-32493480	137.03-138.32	2.25
	qAC3.2	3	33386000-34509821	143.12-147.93	3.23
	qAC3.3	3	35314343-35646346	151.38-152.81	2.30
	qAC5.1	5	22147125-22524107	94.94-96.55	2.88
	qAC11.1	11	9832804-10200412	42.15-43.73	2.28
	qAC12.1	12	927518-2689107	3.98-11.53	2.31
	qAC12.2	12	23013674-23469132	98.65-100.61	2.50

Antioxidant physiological indicators	Gene ID	QTL locus	Functional annotation	Gene name
Hydroxyl radical scavenging rate	LOC_Os06g01850	qHRSR6.1	Leaf-type ferredoxin-NADP⁺oxidoreductase	OsLFNR2 (Da et al., 2023)
	LOC_Os06g48000	qHRSR6.3	Peroxidase precursor
	LOC_Os06g48020	qHRSR6.3	Peroxidase precursor
Total phenolic content	LOC_Os04g45210	qTPC4.1	Peroxisomal biogenesis factor 11
	LOC_Os12g07820	qTPC12.2	Ascorbate peroxidase gene	OsAPx6 (Chou et al., 2012)
	LOC_Os12g07830	qTPC12.2	Ascorbate peroxidase gene	OsAPx5 (Hong et al., 2007)
Flavonoid content	LOC_Os05g12190	qFC5.2	Chalcone synthase
	LOC_Os05g12210	qFC5.2	Chalcone synthase
	LOC_Os05g12240	qFC5.2	Chalcone synthase
	LOC_Os05g45120	qFC5.4	Anthocyanidin 5,3-O-glucosyltransferase
	LOC_Os12g16260	qFC12.3	Isoflavone reductase
Anthocyanin content	LOC_Os03g60509	qAC3.2	Chalcone isomerase gene	OsCHI (Lam et al., 2022)
Anthocyanin content	LOC_Os03g62480	qAC3.3	UV-B-responsive glycosyltransferase; flavonoid 7-O-glycosyltransferase

水稻抗氧化性状遗传位点挖掘及候选基因分析

Genetic Locus Mining and Candidate Gene Analysis of Antioxidant Traits in Rice

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