甜菜HIPPs基因家族鉴定与镉胁迫下的表达分析

doi:10.3724/SP.J.1006.2023.34010

作物学报 ›› 2023, Vol. 49 ›› Issue (12): 3302-3314.doi: 10.3724/SP.J.1006.2023.34010

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

甜菜HIPPs基因家族鉴定与镉胁迫下的表达分析

赵晓鑫¹^,²(), 黄烁淇¹^,², 谭文勃¹^,², 兴旺¹^,², 刘大丽¹^,²^,^*()

¹黑龙江大学国家甜菜种质中期库, 黑龙江哈尔滨 150080
²黑龙江省普通高等学校甜菜遗传育种重点实验室 / 黑龙江大学现代农业与生态环境学院, 黑龙江哈尔滨 150080

收稿日期:2023-01-13 接受日期:2023-05-24 出版日期:2023-12-12 网络出版日期:2023-06-16
通讯作者: * 刘大丽, E-mail: daliliu_hlju@163.com* E-mail: daliliu_hlju@163.com
作者简介:E-mail: 953511214@qq.com
基金资助:
国家自然科学基金青年基金项目(31606229);黑龙江省自然科学基金项目(LH2019C057);财政部和农业农村部国家现代农业产业技术体系建设专项(Sugar, CARS-170102);农业农村部项目(19221878);农业农村部项目(19211031);农业农村部项目(19210911);黑龙江省普通本科高等学校青年创新人才培养计划项目(UNPYSCT-2020)

Identification and relative expression profile of HIPPs gene family cadmium stress in sugar beet

ZHAO Xiao-Xin¹^,²(), HUANG Shuo-Qi¹^,², TAN Wen-Bo¹^,², XING Wang¹^,², LIU Da-Li¹^,²^,^*()

¹National Beet Germplasm Mid-term Bank, Heilongjiang University, Harbin 150080, Heilongjiang, China
²Key Laboratory of Beet Genetics and Breeding / College of Modern Agriculture and Ecological Environment, Heilongjiang University, Harbin 150080, Heilongjiang, China

Received:2023-01-13 Accepted:2023-05-24 Published:2023-12-12 Published online:2023-06-16
Supported by:
Youth Program of National Natural Science Foundation of China(31606229);Natural Science Foundation of Heilongjiang Province(LH2019C057);China Agriculture Research System of MOF and MARA(Sugar, CARS-170102);Ministry of Agriculture and Rural Affairs Project(19221878);Ministry of Agriculture and Rural Affairs Project(19211031);Ministry of Agriculture and Rural Affairs Project(19210911);Innovative Training Plan for Young Talents in Heilongjiang Ordinary Undergraduate Colleges and Universities(UNPYSCT-2020)

摘要/Abstract

摘要：

镉离子平衡和解毒调控是探索甜菜镉胁迫耐受机制的核心, 也是利用甜菜进行重金属生物修复的基础。重金属相关的异戊二烯植物蛋白(HIPPs)是一类多功能金属伴侣蛋白, 在镉离子吸收、转运及区隔化中发挥关键作用。前期甜菜响应镉胁迫的转录组表达谱研究中, 发现BvHIPPs存在差异表达。基于此, 本研究通过生物信息学方法全基因组鉴定了BvHIPPs基因家族成员, 并对其理化性质、进化关系、基因结构、顺式作用元件、染色体定位及在镉胁迫下的转录表达特性进行了深入的分析。结果表明, 甜菜基因组中共有23个BvHIPPs家族成员, 均含有HMA结构域和异戊二烯化基序, 其中16个BvHIPPs被定位于细胞核。顺式作用元件分析发现BvHIPPs可参与多种生物与非生物胁迫响应。转录组数据表明23个BvHIPPs均不同程度的参与到甜菜对镉胁迫的应答过程, 并且进一步的qRT-PCR分析验证了BvHIPPs应答镉胁迫的调控特点。结果表明, BvHIPPs可能在甜菜适应镉胁迫的过程中发挥着重要作用, 研究结果为甜菜在重金属污染生物修复的分子机制研究奠定基础。

关键词: 甜菜, HIPPs, 镉胁迫, 基因家族鉴定, 转录表达特性

Abstract:

Cadmium (Cd) ion balance and detoxification regulation are the cores of exploring cadmium tolerance mechanism in sugar beet, and the basis of heavy metal bioremediation using sugar beet. Heavy metal-associated isoprene plant proteins (HIPPs) are a class of multifunctional metal chaperone proteins, which may play a key role in absorption, transport, and compartmentalization of Cd ions. In the transcriptome expression profile of sugar beet responding to cadmium stress, BvHIPPs were found to be differentially expressed. Based on the above results, the whole BvHIPPs gene family members in beet genome was identified by bioinformatics method, and their physicochemical properties, evolutionary relationships, gene structure, cis-acting elements, chromosome localization, transcription, and expression characteristics under cadmium stress were analyzed. The results showed that there were 23 BvHIPPs family members in the beet genome, all of which contained HMA domains and isoprenylation motif, and 16 BvHIPPs were predicted to be located in the nucleus. According to cis-acting element analysis, beet BvHIPPs can participate in a variety of biological and abiotic stress responses. Transcriptomic analysis showed that all 23 BvHIPPs differentially participated in sugar beet in response to Cd, and qRT-PCR analysis verified furtherly the regulatory characteristics of the BvHIPPs correlated with Cd response. BvHIPPs might play an essential role in the adaptation of sugar beet to cadmium stress. The results suggest that BvHIPPs may play an important role in the process of beet adaptation to cadmium stress, and the results will lay a foundation for the molecular mechanism of beet bioremediation in heavy metal pollution.

Key words: sugar beet, HIPPs, cadmium stress, gene family identification, transcriptional expression characteristics

赵晓鑫, 黄烁淇, 谭文勃, 兴旺, 刘大丽. 甜菜HIPPs基因家族鉴定与镉胁迫下的表达分析[J]. 作物学报, 2023, 49(12): 3302-3314.

ZHAO Xiao-Xin, HUANG Shuo-Qi, TAN Wen-Bo, XING Wang, LIU Da-Li. Identification and relative expression profile of HIPPs gene family cadmium stress in sugar beet[J]. Acta Agronomica Sinica, 2023, 49(12): 3302-3314.

图/表 8

表1

表2

BvHIPPs基因家族的基本信息"

基因 Gene name	氨基酸数 Amino acid number	分子量 Relative molecular mas (kD)	等电点 Isoelectric point	不稳定系数 Instability index	脂肪系数 Aliphatic index	平均亲水性 Hydrophilic coefficient	亚细胞定位 Subcellular localization
BVRB_6g129220	131	14.52	6.76	39.11	72.82	-0.355	未定位Not located
BVRB_7g161540	355	38.77	9.06	32.26	50.45	-1.251	细胞核 Nucleus
BVRB_5g126430	343	38.33	6.00	48.78	51.40	-1.301	细胞核 Nucleus
BVRB_3g050800	340	37.64	4.91	71.30	65.06	-0.926	细胞核 Nucleus
BVRB_2g043150	149	16.82	9.45	43.45	71.81	-0.556	细胞核 Nucleus
BVRB_2g027600	147	16.52	9.28	50.11	66.26	-0.559	细胞核 Nucleus
BVRB_6g135550	155	17.25	9.42	33.95	64.71	-0.561	未定位Not located
BVRB_1g021330	147	16.38	9.39	17.54	65.65	-0.416	线粒体 Mitochondrion
BVRB_9g214040	158	18.07	9.01	40.36	65.82	-0.676	线粒体, 细胞核 Mitochondrion, nucleus
BVRB_5g104360	143	16.47	9.17	34.27	70.77	-0.617	细胞核 Nucleus
BVRB_3g055490	134	14.84	6.28	36.58	72.84	-0.249	未定位Not located
BVRB_4g081750	508	53.07	9.35	47.01	32.62	-1.273	细胞核 Nucleus
BVRB_3g069170	562	55.92	9.85	49.7	34.63	-0.894	细胞核 Nucleus
BVRB_7g164120	417	44.63	8.11	67.82	58.56	-0.823	细胞核 Nucleus
BVRB_1g004050	167	19.32	7.60	52.93	65.93	-1.255	细胞核 Nucleus
BVRB_4g075740	345	37.67	6.19	52.19	45.16	-1.208	细胞核 Nucleus
BVRB_5g122000	137	14.99	9.05	41.58	73.14	-0.381	细胞核 Nucleus
BVRB_8g184430	297	34.30	6.62	56.84	62.59	-1.118	细胞核 Nucleus
BVRB_3g053290	132	14.88	8.85	40.62	73.03	-0.767	细胞核 Nucleus
BVRB_3g053300	127	14.16	8.28	27.07	94.88	-0.181	细胞核 Nucleus
BVRB_6g142690	101	11.21	9.67	48.14	100.99	-0.149	叶绿体, 细胞核 Chloroplast, nucleus
BVRB_3g048110	370	41.00	5.70	76.65	42.16	-1.253	细胞核 Nucleus
BVRB_3g053310	113	12.70	8.37	24.08	81.77	-0.295	叶绿体, 细胞质, 细胞核Chloroplast, cytoplasm, nucleus

表2

图1

图2

图3

图4

图5

图6

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基因Gene name	上游引物Forward primer (5°-3°)	下游引物Reverse primer (5°-3°)
BVRB_1g021330	AAGCAACAGAAGGTGACGGT	GGGTGTGCCACCATAGTGTA
BVRB_3g050800	ATAAAAGGCGTGGTGGAGCC	GGGTTCACCCTCATTTGCCG
BVRB_4g075740	AACACGGCGTATCCTAGCAG	TATCAGACGGTGGAACTGCG
BVRB_6g135550	GCAAAAGCCACAGGGAAGAAG	GTCCACCTTCCTGACATGACC
BVRB_3g053310	CTCAGTTGGGCCAGCAAAAG	GTACGGAGGGTTGTAGCCG
BVRB_6g129220	GCCCGATCTGAAGTGTCCAT	CTATGGGCCTGCCATCACAA
BVRB_7g161540	TTGTCCGCAAGTGTGATGGA	CCTTCTTTGCAGGAGCAAGC
BVRB_5g126430	TCCTATTCCGAAACCACCCG	GCATGTGAACCTTGAGCACC
BVRB_2g043150	TGTGGAGCCAAATAGGGTGC	TTTGACATAGCCGGAGGGTG
BVRB_2g027600	ACAGGGCACGTAGATCCAAA	GCCAGCAGGTGCTCTCTTAT
BVRB_9g214040	CGTGTGAAGCATCGAACAGG	ACTCCAGGAGCGTAAGGGT
BVRB_5g104360	GTTCTGGCCTTACCCTAACCC	GAGGGTGCCATGTCTATCACC
BVRB_3g055490	ATCAAAAGGGCAGGCAAAGC	CCGTCGGAAGTGTCGTAGTAA
BVRB_4g081750	TTGCCAGCTTCCCACATGAT	TGCGGATTCATCGGTACTGG
BVRB_3g069170	AAATGGGCCCTATGGGTTCG	TGGAAGTATTCGGGTGCTGC
BVRB_7g164120	CTACCCGCACAGGCCATATC	GTTGCTACTGCTCGATTGCG
BVRB_1g004050	AGGCGAAGCAGAAAGCTCTTA	TGTTGTTGGCCAATACTTGCG
BVRB_5g122000	AGGTTGAGCCAGTGCCAATC	ACTGCCTCTGTTGCTGATCG
BVRB_8g184430	TGTGGCGGTGGATCGAAAA	GCATTTCGGCCCGTTTTGT
BVRB_3g053290	CACAGAAATTGTCTCAGTTGGGC	GAAAGCCCGTTACAATCTCGC
BVRB_3g053300	TTGGCGATGTGGATCCTGTAG	TTTGCAGGCCCAACTGAGAC
BVRB_6g142690	TGACCGGAAATGTGGAACCAA	TGAGCAACGTAAGGGTAGGAC
BVRB_3g048110	CGTAGTATACCACCATCCGCC	TTGATGGCTGCTCCAGTGTC
BvGAPDH	GCTTTGAACGACCACTTCGC	ACGCCGAGAGCAACTTGAAC

甜菜HIPPs基因家族鉴定与镉胁迫下的表达分析

Identification and relative expression profile of HIPPs gene family cadmium stress in sugar beet

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