芥菜HMA家族基因鉴定及其在镉胁迫下的表达分析

doi:10.16420/j.issn.0513-353x.2022-0509

摘要/Abstract

摘要：

芥菜（Brassica juncea）对多种重金属具有富集能力。重金属ATP酶（heavy metal transporting ATPase，HMA）在植物转运重金属过程中具有重要的作用。基于基因组和转录组数据，对芥菜HMA家族基因成员进行了全基因组鉴定。芥菜基因组中包含27个HMA基因，其编码的蛋白质中有6个不稳定指数大于40，有22个为疏水性蛋白；这些基因聚类为P-1B-1、P-1B-2和P-1B-4等3个亚家族；27个HMA蛋白均为膜蛋白，都具有E1-E2 ATPase和hydrolase结构域；芥菜HMA蛋白都含有8 ~ 15个保守基序，不同亚族因具有独特的保守基序结构从而转运重金属的种类不同；在芥菜HMA基因启动子区域中与生长发育有关的顺式作用元件TGA和与逆境响应相关的顺式作用元件ABRE、MBS、LTR、TC广泛分布；在镉胁迫下，BjuA033764、BjuB019118、BjuB035256、BjuB045293等基因在叶片中表达量明显上调，BjuA003596、BjuB040613、BjuA025022等基因在根系中表达量明显上调，说明其参与了芥菜对镉胁迫的响应。

关键词: 芥菜, 重金属ATP酶, HMA基因家族, 镉胁迫, 结构域, 顺式作用元件

Abstract:

Brassica juncea can enrich a variety of heavy metals. Heavy metal transporting ATPase（HMA）plays an important role in the process of transporting heavy metals in plants. Based on genomic and transcriptomic data，the HMA gene family members in B. juncea were identified. A total of 27 HMA genes were found in B. juncea genome. Six of these HMA proteins have instability indexes greater than 40. Twenty-two members are hydrophobic proteins. These genes cluster into three subfamilies，P-1B-1，P-1B-2，and P-1B-4. All the HMAs are membrane proteins containing the E1-E2 ATPase and hydrolase domains. Each HMA protein contains 1-8 motifs. Different subfamilies have unique conserved motifs according to the types of heavy metals transported by them. In the promoter regions of the HMA genes，the cis-acting element TGA related to the regulation of growth and development，the cis-acting elements ABRE，MBS，LTR，and TC related to the regulation of stress response are widely distributed. Under cadmium（Cd）stress，BjuA033764，BjuB019118，BjuB035256，and BjuB045293 were up-regulated in leaves，and BjuA003596，BjuB040613，and BjuA025022 were up-regulated in roots，indicating their involvement in the response to Cd stress.

Key words: Brassica juncea, heavy metal transporting ATPase, HMA gene family, cadmium stress, domain, cis-acting element

中图分类号:

S637

黄治皓, 刘婷婷, 董旭杰, 严明理, 刘志祥, 曾超珍. 芥菜HMA家族基因鉴定及其在镉胁迫下的表达分析[J]. 园艺学报, 2023, 50(6): 1230-1242.

HUANG Zhihao, LIU Tingting, DONG Xujie, YAN Mingli, LIUZhixiang , ZENG Chaozhen. Identification and Expression Analysis of HMA Gene Family in Brassica juncea Under Cadmium Stress[J]. Acta Horticulturae Sinica, 2023, 50(6): 1230-1242.

图/表 9

参考文献 34

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基因ID Gene ID	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer
BjuB002000	ATCGTTGGCTTCCCTCTCGTT	ACATCCACCATTGACCGGCTTG
BjuA003596	AAGAACCGTCATCGTCGTCCA	AGAAGTACGCCGGAAACCAC
BjuB040613	TCATCGTCGTCCATGATAGCC	AATTAGTTTCTCCGGTTACCCTC
BjuO003139	CGATACTTGCCAAAGCCGTTG	TGCATAACCGCATTCGCCTT
BjuA025267	GCCTCCTCATCTCTCCGTCCC	TTTGCCGTCCACTTTCACGTT
BjuA042408	CAAGAACCGTCATCGTTGTCC	GCAACAAGAGCGAACCACT
Actin	GAATCCACGAGACGACTTACAAC	CGATCCAGACACTGTACTTCCTC

基因ID Gene ID	正向引物（5′-3′） Forward primer	反向引物（5′-3′） Reverse primer
BjuB002000	ATCGTTGGCTTCCCTCTCGTT	ACATCCACCATTGACCGGCTTG
BjuA003596	AAGAACCGTCATCGTCGTCCA	AGAAGTACGCCGGAAACCAC
BjuB040613	TCATCGTCGTCCATGATAGCC	AATTAGTTTCTCCGGTTACCCTC
BjuO003139	CGATACTTGCCAAAGCCGTTG	TGCATAACCGCATTCGCCTT
BjuA025267	GCCTCCTCATCTCTCCGTCCC	TTTGCCGTCCACTTTCACGTT
BjuA042408	CAAGAACCGTCATCGTTGTCC	GCAACAAGAGCGAACCACT
Actin	GAATCCACGAGACGACTTACAAC	CGATCCAGACACTGTACTTCCTC

基因Gene	结构域Domain			基因Gene	结构域Domain
基因Gene	HMA	E1-E2 ATPase	Hydrolase	基因Gene	HMA	E1-E2 ATPase	Hydrolase
AtHMA1	-	E1-E2 ATPase	Hydrolase	BjuO003139	-	E1-E2 ATPase	Hydrolase
AtHMA2	-	E1-E2 ATPase	Hydrolase	BjuB035256	-	E1-E2 ATPase	Hydrolase
AtHMA3	-	E1-E2 ATPase	Hydrolase	BjuA023298	-	E1-E2 ATPase	Hydrolase
AtHMA4	-	E1-E2 ATPase	Hydrolase	BjuA025267	-	E1-E2 ATPase	Hydrolase
AtHMA5	[HMA]₃	E1-E2 ATPase	Hydrolase	BjuA042408	HMA	E1-E2 ATPase	Hydrolase
AtHMA6	HMA	E1-E2 ATPase	Hydrolase	BjuB028207	[HMA]₂	E1-E2 ATPase	Hydrolase
AtHMA7	[HMA]₂	E1-E2 ATPase	Hydrolase	BjuA046861	[HMA]₂	E1-E2 ATPase	Hydrolase
AtHMA8	HMA	E1-E2 ATPase	Hydrolase	BjuB035323	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB002000	-	E1-E2 ATPase	Hydrolase	BjuA038121	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB034828	-	E1-E2 ATPase	Hydrolase	BjuB019118	HMA	E1-E2 ATPase	Hydrolase
BjuA036645	-	E1-E2 ATPase	Hydrolase	BjuB027969	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuA002855	-	E1-E2 ATPase	Hydrolase	BjuA025022	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuO011640	HMA	E1-E2 ATPase	Hydrolase	BjuB036631	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuA003595	-	E1-E2 ATPase	Hydrolase	BjuA033764	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB040614	-	E1-E2 ATPase	Hydrolase	BjuA003262	HMA	E1-E2 ATPase	Hydrolase
BjuA003596	HMA	E1-E2 ATPase	Hydrolase	BjuA038847	HMA	E1-E2 ATPase	Hydrolase
BjuB040613	HMA	E1-E2 ATPase	Hydrolase	BjuB045293	HMA	E1-E2 ATPase	Hydrolase
BjuA042404	[HMA]₃	E1-E2 ATPase	Hydrolase

基因Gene	结构域Domain			基因Gene	结构域Domain
基因Gene	HMA	E1-E2 ATPase	Hydrolase	基因Gene	HMA	E1-E2 ATPase	Hydrolase
AtHMA1	-	E1-E2 ATPase	Hydrolase	BjuO003139	-	E1-E2 ATPase	Hydrolase
AtHMA2	-	E1-E2 ATPase	Hydrolase	BjuB035256	-	E1-E2 ATPase	Hydrolase
AtHMA3	-	E1-E2 ATPase	Hydrolase	BjuA023298	-	E1-E2 ATPase	Hydrolase
AtHMA4	-	E1-E2 ATPase	Hydrolase	BjuA025267	-	E1-E2 ATPase	Hydrolase
AtHMA5	[HMA]₃	E1-E2 ATPase	Hydrolase	BjuA042408	HMA	E1-E2 ATPase	Hydrolase
AtHMA6	HMA	E1-E2 ATPase	Hydrolase	BjuB028207	[HMA]₂	E1-E2 ATPase	Hydrolase
AtHMA7	[HMA]₂	E1-E2 ATPase	Hydrolase	BjuA046861	[HMA]₂	E1-E2 ATPase	Hydrolase
AtHMA8	HMA	E1-E2 ATPase	Hydrolase	BjuB035323	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB002000	-	E1-E2 ATPase	Hydrolase	BjuA038121	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB034828	-	E1-E2 ATPase	Hydrolase	BjuB019118	HMA	E1-E2 ATPase	Hydrolase
BjuA036645	-	E1-E2 ATPase	Hydrolase	BjuB027969	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuA002855	-	E1-E2 ATPase	Hydrolase	BjuA025022	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuO011640	HMA	E1-E2 ATPase	Hydrolase	BjuB036631	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuA003595	-	E1-E2 ATPase	Hydrolase	BjuA033764	[HMA]₂	E1-E2 ATPase	Hydrolase
BjuB040614	-	E1-E2 ATPase	Hydrolase	BjuA003262	HMA	E1-E2 ATPase	Hydrolase
BjuA003596	HMA	E1-E2 ATPase	Hydrolase	BjuA038847	HMA	E1-E2 ATPase	Hydrolase
BjuB040613	HMA	E1-E2 ATPase	Hydrolase	BjuB045293	HMA	E1-E2 ATPase	Hydrolase
BjuA042404	[HMA]₃	E1-E2 ATPase	Hydrolase

亚族 Subgroup	基因 Gene	长度/aa Length	分子量/kD MW	等电点 pI	不稳定指数Instability index	疏水性指数 GRAVY	亚细胞定位 Subcellular location
亚族 Subgroup	基因 Gene	长度/aa Length	分子量/kD MW	等电点 pI	不稳定指数Instability index	疏水性指数 GRAVY	WoLF PSORT	BUSCA
P-1B-1	BjuB028207	998	108.48	5.72	35.69	0.16	PM	PM
	BjuA046861	1 184	129.44	5.62	35.11	0.08	PM	PM
	BjuB019118	839	90.81	5.55	27.34	0.28	PM	PM
	BjuB035323	978	106.55	5.94	28.84	0.17	PM	PM
	BjuA038121	973	105.65	6.13	26.97	0.19	PM	PM
	BjuB027969	1 006	107.54	5.02	32.02	0.22	PM	PM
	BjuA033764	980	104.40	4.93	31.54	0.23	PM	PM
	BjuA025022	999	106.84	5.00	33.47	0.25	PM	PM
	BjuB036631	1 000	106.96	5.01	33.48	0.25	PM	PM
	BjuA003262	938	98.60	8.76	41.58	0.11	PM	MM
	BjuA038847	885	94.22	5.68	37.54	0.16	PM	MM
P-1B-2	BjuA002855	683	73.81	7.23	33.38	0.06	PM	MM
	BjuO011640	801	86.29	7.10	36.86	0.22	PM	PM
	BjuA003595	758	81.39	5.98	33.75	0.26	PM	PM
	BjuB040614	760	81.84	5.88	33.83	0.25	PM	PM
	BjuA003596	872	94.33	6.93	35.83	0.04	PM	PM
	BjuB040613	928	99.86	6.78	35.05	0.04	PM	PM
	BjuA042404	903	97.14	7.97	32.33	0	PM	PM
	BjuO003139	1 271	137.78	7.79	84.05	−0.24	PM	PM
	BjuB035256	1 220	132.84	6.23	43.35	−0.19	PM	PM
	BjuA023298	1 193	129.78	6.82	41.95	−0.23	PM	PM
	BjuA025267	1 162	125.17	6.31	91.34	−0.08	PM	PM
	BjuA042408	795	85.53	6.80	38.11	0.25	PM	PM
	BjuB045293	613	63.83	9.29	46.64	0.22	PM	MM
P-1B-4	BjuB002000	820	88.05	7.87	35.75	0.12	PM	MM
	BjuB034828	783	84.77	6.86	35.14	0.18	PM	OM
	BjuA036645	775	84.25	6.74	34.78	0.13	PM	OM