Identification and Expression Analysis of HMA Gene Family in Brassica juncea Under Cadmium Stress

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

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (6): 1230-1242.doi: 10.16420/j.issn.0513-353x.2022-0509

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Identification and Expression Analysis of HMA Gene Family in Brassica juncea Under Cadmium Stress

HUANG Zhihao¹, LIU Tingting¹, DONG Xujie¹, YAN Mingli², LIUZhixiang ¹, ZENG Chaozhen¹^,^*()

¹Hunan Provincial Key Laboratory of Forestry Biotechnology，College of Life Science and Technology，Central South University of Forestry and Technology，Changsha 410004，China
²Crop Research Institute，Hunan Academy of Agricultural Sciences，Changsha 410125，China

Received:2022-10-21 Revised:2023-03-14 Online:2023-06-25 Published:2023-06-27
Contact: * （E-mail：chaozhenzeng@163.com）

Abstract

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

CLC Number:

S637

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.

Figures/Tables 9

References 34

[1]	Arguëllo J M. 2003. Identification of ion-selectivity determinants in heavy-metal transport P1B-type ATPases. Molecular Membrane Biology, 195 (2):93-108.
[2]	Axelsen K B, Palmgren M G. 2001. Inventory of the superfamily of P-type ion pumps in Arabidopsis. Plant Physiology, 126 (2):696-706. pmid: 11402198
[3]	Choudhury M R, Islam M S, Ahmed Z U, Nayar F. 2016. Phytoremediation of heavy metal contaminated buriganga riverbed sediment by Indian mustard and marigold plants. Environmental Progress & Sustainable Energy, 35 (1):117-124.
[4]	Clemens S. 2001. Molecular mechanisms of plant metal tolerance and homeostasis. Planta, 212 (4):475-486. doi: 10.1007/s004250000458 pmid: 11525504
[5]	Cola N A, Sancenon V, Navarro S R, Mayo S, Thiele D J, Ecker J R, Puig S, Penarrubia L. 2006. The Arabidopsis heavy metal P-type ATPase HMA 5 interacts with metallochaperones and functions in copper detoxification of roots. Plant Journal, 45 (2):225-236. doi: 10.1111/tpj.2006.45.issue-2 URL
[6]	Courbot M, Willems G, Motte P, Arvidsson S, Roosens N, Laprade P S, erbruggen N V. 2007. A major quantitative trait locus for cadmium tolerance in Arabidopsis halleri colocalizes with HMA4,a gene encoding a heavy metal ATPase. Plant Physiology, 144 (2):1052-1065. doi: 10.1104/pp.106.095133 pmid: 17434989
[7]	Ghany S E A, Moule P M, Niyogi K K, Pilon M, Shikanai T. 2005. Two P-type ATPases are required for copper delivery in Arabidopsis thaliana chloroplasts. Plant Cell, 17 (4):1233-1251. doi: 10.1105/tpc.104.030452 URL
[8]	Goswami S, Das S. 2015. A study on cadmium phytoremediation potential of indian mustard,Brassica juncea. International Journal of Phytoremediation, 17 (6):583-588. doi: 10.1080/15226514.2014.935289 URL
[9]	Gravot A, Lieutaud A, Verret F, Auroy P, Vavasseur A, Richaud P. 2004. AtHMA3,a plant P1B-ATPase,functions as a Cd/Pb transporter in yeast. Febs Letters, 561 (1-3):22-28. doi: 10.1016/S0014-5793(04)00072-9 URL
[10]	Hanikenne M, Talke I N, Haydon M J, Lanz C, Nolte A, Motte P, Kroymann J, Weigel D, Kramer U. 2008. Evolution of metal hyperaccumulation required cis-regulatory changes and triplication of HMA4. Nature, 453 (7193):391-395. doi: 10.1038/nature06877
[11]	Hussain D, Haydon M J, Wang Y, Wong E, Sherson S M, Young J, Camakaris J, Harper J F, Cobbett C S. 2004. P-type ATPase heavy metal transporters with roles in essential zinc homeostasis in Arabidopsis. Plant Cell, 16 (5):1327-1339. doi: 10.1105/tpc.020487 pmid: 15100400
[12]	Kim Y Y, Choi H, Segami S, Cho H T, Martinoia E, Masayoshi, Maeshima, Lee Y. 2009. AtHMA1 contributes to the detoxification of excess Zn(II) in Arabidopsis. Plant Journal, 58 (5):737-753. doi: 10.1111/tpj.2009.58.issue-5 URL
[13]	Liao Tian-lu, Liao Tian-jiang, Lü Lin-hua. 2015. Application of phytoremediation technology in heavy metal contaminated soil. Beijing Agriculture, 6 (17):192. (in Chinese)
	廖天录, 廖天江, 吕麟华. 2015. 植物修复技术在重金属污染土壤中的应用. 北京农业, 6 (17):192.
[14]	Liu L, Yin H, Liu Y, Shen L, Yang X, Zhang D, Li M, Yan M. 2021. Analysis of cadmium-stress-induced microRNAs and their targets reveals bra-miR172b-3p as a potential Cd²⁺-specific resistance factor in Brassica juncea. Processes, 9 (7):1099. doi: 10.3390/pr9071099 URL
[15]	Miyadate H, Adachi S, Hiraizumi A, Tezuka K, Nakazawa N, Kawamoto T, Katou K, Kodama I, Sakurai K, Takahashi H, Nagasawa N S, Watanabe A, Fujimura T, Akagi H. 2011. OsHMA3,a P1B-type of ATPase affects root-to-shoot cadmium translocation in rice by mediating efflux into vacuoles. New Phytologist, 189 (1):190-199. doi: 10.1111/nph.2010.189.issue-1 URL
[16]	Morel M, Crouzet J, Gravot A, Auroy P, Leonhardt N, Vavasseur A, Richaud P. 2009. AtHMA3,a P1B-ATPase allowing Cd/Zn/Co/Pb vacuolar storage in Arabidopsis. Plant Physiology, 149 (2):894-904. doi: 10.1104/pp.108.130294 URL
[17]	Mourato M P, Moreira I N, Leitao I, Pinto F R, Sales J R, Martins L L. 2015. Effect of heavy metals in plants of the genus Brassica. International Journal of Molecular Sciences, 16 (8):17975-17998. doi: 10.3390/ijms160817975 pmid: 26247945
[18]	Palmgren M G, Clemens S, Williams L E, Kramer U, Borg S, Schjorring J K, Sanders D. 2008. Zinc biofortification of cereals:problems and solutions. Trends In Plant Science, 13 (9):464-473. doi: 10.1016/j.tplants.2008.06.005 pmid: 18701340
[19]	Papoyan A, Kochian L V. 2004. Identification of Thlaspi caerulescens genes that may be involved in heavy metal hyperaccumulation and tolerance Characterization of a novel heavy metal transporting ATPase. Plant Physiology, 136 (3):3814-3823. pmid: 15516513
[20]	Parkin I A P, Gulden S M, Sharpe A G, Lukens L, Trick M, Osborn T C, Lydiate D J. 2005. Segmental structure of the Brassica napus genome based on comparative analysis with Arabidopsis thaliana. Genetics, 171 (2):765-781. doi: 10.1534/genetics.105.042093 URL
[21]	Puig S, Colas N A, Molina A G, Pearrubia L. 2007. Copper and iron homeostasis in Arabidopsis:responses to metal deficiencies,interactions and biotechnological applications. Plant Cell and Environment, 30 (3):271-290. doi: 10.1111/j.1365-3040.2007.01642.x URL
[22]	Sankaran R P, Ebbs S D. 2008. Transport of Cd and Zn to seeds of Indian mustard(Brassica juncea)during specific stages of plant growth and development. Physiologia Plantarum, 132 (1):69-78.
[23]	Sun Tao, Zhang Yu-xiu, Chai Tuan-yao. 2011. Research progress on tolerance of Indian mustard(Brassica juncea L.) to heavy metal. Chinese Journal of Eco-Agriculture, 19 (1):226-234. (in Chinese)
	孙涛, 张玉秀, 柴团耀. 2011. 印度芥菜(Brassica juncea L.)重金属耐性机理研究进展. 中国生态农业学报, 19 (1):226-234.
[24]	Tan P, Zeng C, Wan C, Liu Z, Dong X, Peng J, Lin H, Li M, Liu Z, Yan M. 2021. Metabolic profiles of Brassica juncea roots in response to cadmium stress. Metabolites, 11 (6):383. doi: 10.3390/metabo11060383 URL
[25]	Verret F, Gravot A, Auroy P, Preveral S, Forestier C, Vavasseur A, Richaud P. 2005. Heavy metal transport by AtHMA4 involves the N-terminal degenerated metal binding domain and the C-terminal His11 stretch. Febs Letters, 579 (6):1515-1522. doi: 10.1016/j.febslet.2005.01.065 URL
[26]	Verreta F, Gravota A, Auroya P, Leonhardta N, Davidb P, Nussaumeb L, Vavasseura A, Richaud P. 2004. Overexpression of AtHMA4enhances root-to-shoot translocation of zinc and cadmium and plant metal tolerance. Febs Letters, 576 (3):306-312. doi: 10.1016/j.febslet.2004.09.023 URL
[27]	Wang Ruonan, Nie Lanchun, Zhang Shuangshuang, Cui Qiang, Jia Mingfei. 2019. Research progress on plant resistance to heavy metal stress. Acta Horticulturae Sinica, 46 (1):157-170. (in Chinese) doi: 10.16420/j.issn.0513-353x.2018-0585 URL
	王若男, 乜兰春, 张双双, 崔强, 贾明飞. 2019. 植物抗重金属胁迫研究进展. 园艺学报, 46 (1):157-170. doi: 10.16420/j.issn.0513-353x.2018-0585 URL
[28]	Wang Xiao-tong, Li Hao-yang, Xu Ji-chen. 2014. Bioinformatics analysis of the heavy metal transporting ATPase gene family in poplar genome. Plant Physiology Journal, 50 (7):891-900. (in Chinese)
	王晓桐, 李昊阳, 徐吉臣. 2014. 毛果杨HMA基因家族的生物信息学分析. 植物生理学报, 50 (7):891-900.
[29]	Williams L E, Mills R F. 2005. P(1B)-ATPases--an ancient family of transition metal pumps with diverse functions in plants. Trends In Plant Science, 10 (10):491-502. doi: 10.1016/j.tplants.2005.08.008 pmid: 16154798
[30]	Wong C K E, Cobbett C S. 2009. HMA P-type ATPases are the major mechanism for root-to-shoot Cd translocation in Arabidopsis thaliana. New Phytologist, 181 (1):71-78. doi: 10.1111/nph.2009.181.issue-1 URL
[31]	Wong C K E, Jarvis R S, Sherson S M, Cobbett C S. 2009. Functional analysis of the heavy metal binding domains of the Zn/Cd-transporting ATPase,HMA2,in Arabidopsis thaliana. New Phytologist, 181 (1):79-88. doi: 10.1111/nph.2009.181.issue-1 URL
[32]	Xu D, Lu Z C, Jin K M, Qiu W M, Qiao G R, Han X J, Zhuo R Y. 2020. SPDE:a multi-functional software for sequence processing and data extraction. Bioinformatics, 37 (20):3686-3687. doi: 10.1093/bioinformatics/btab235 URL
[33]	Zhang Fu-gui, Xiao Xin, Yan Gui-xin. 2017. Evolution of HMA gene family in Brassica AC genomes. Chinese Journal of Oil Crop Sciences, 39 (3):294-307. (in Chinese)
	张付贵, 肖欣, 闫贵欣. 2017. HMA基因家族在芸薹属AC基因组中的进化分析. 中国油料作物学报, 39 (3):294-307.
[34]	Zhang Yu-xiu, Chai Tuan-yao. 2006. Isolation and function of heavy metal responsive gene in plant. Beijing: China Agriculture Press. (in Chinese)
	张玉秀, 柴团耀. 2006. 植物重金属调节基因的分离和功能. 北京: 中国农业出版社.

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

Identification and Expression Analysis of HMA Gene Family in Brassica juncea Under Cadmium Stress

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 9

References 34

Related Articles 9

Recommended Articles

Metrics

Comments

亚族 Subgroup	基因 Gene	染色体 Chr	顺式作用元件数量 Number of cis-acting element
亚族 Subgroup	基因 Gene	染色体 Chr	G-Box	ABRE	LTR	TC	MBS	TGA
P-1B-1	AtHMA5	1	1	1	0	2	0	2
	AtHMA6	4	0	1	0	1	1	1
	AtHMA7	5	1	1	0	3	1	1
	AtHMA8	5	0	1	0	1	2	1
	BjuB028207	B04	0	1	0	0	0	1
	BjuA046861	A09	0	0	0	0	0	0
	BjuB019118	B08	0	0	0	0	1	0
	BjuB035323	B02	0	0	0	1	0	0
	BjuA038121	A10	0	0	0	2	0	0
	BjuB027969	B04	0	0	1	3	1	0
	BjuA033764	A09	0	0	1	3	1	0
	BjuA025022	A06	1	1	0	0	1	0
	BjuB036631	B02	1	1	0	0	1	0
	BjuA003262	A01	0	0	1	0	0	0
	BjuA038847	A10	7	7	2	0	1	0
P-1B-2	AtHMA2	4	1	0	0	1	2	0
	AtHMA3	4	1	0	1	0	2	2
	AtHMA4	2	0	1	1	1	0	3
	BjuA002855	A01	0	0	0	0	1	0
	BjuO011640	Super_scaffold_184_753382_1581011	1	1	0	0	1	2
	BjuA003595	A01	0	0	0	0	0	0
	BjuB040614	B05	1	1	0	2	0	0
	BjuA003596	A01	2	2	2	1	0	3
	BjuB040613	B05	2	2	0	0	0	0
	BjuA042404	A03	2	3	2	0	1	0
	BjuO003139	Contig2023	2	2	1	0	0	0
	BjuB035256	B02	3	1	3	1	0	1
	BjuA023298	A06	1	1	0	2	0	0
	BjuA025267	A07	1	1	3	0	0	0
	BjuA042408	A03	2	0	0	1	1	1
	BjuB045293	B05	0	0	0	0	0	0
P-1B-4	AtHMA1	4	0	0	1	2	2	2
	BjuB002000	B05	1	1	0	0	1	1
	BjuB034828	B02	0	0	0	0	0	0
	BjuA036645	A09	1	1	0	0	1	1

[1]	GAO Pengfei, GAO Bing, FENG Zhenghong, WU Jianhui. Cloning and Cd-resistant Analysis of PsWRKY40 in Potentilla sericea [J]. Acta Horticulturae Sinica, 2023, 50(6): 1269-1283.
[2]	ZHANG Kun, SI Binbin, ZHOU Jun, REN Yufeng, ZHANG Xin, XU Wendi, WANG Jiawei, QIAO Shuai, WANG Huiran. Construction of cDNA Library of Apple Rootstock‘Qingzhen 1’Leaf and Screen of MdMLO Genes’ Upstream Regulator [J]. Acta Horticulturae Sinica, 2023, 50(5): 933-946.
[3]	CHEN Daozong, LIU Yi, SHEN Wenjie, ZHU Bo, TAN Chen. Identification and Analysis of PAP1/2 Homologous Genes in Brassica rapa,B. oleracea and B. napus [J]. Acta Horticulturae Sinica, 2022, 49(6): 1301-1312.
[4]	YANG Tianchen, CHEN Xiaotong, LÜ Ke, ZHANG Di. Expression Pattern and Regulation Mechanism of ApSK₃ Dehydrin （Agapanthus praecox）Response to Abiotic Stress and Hormone Signals [J]. Acta Horticulturae Sinica, 2021, 48(8): 1565-1578.
[5]	BIAN Kun1,2，SHAO Jiarong2，XU Liwei2，SHEN Ziming1,2，CHEN Wei2，and YANG Zhenfeng2,*. Bioinformatic and Expression Analysis of Fasciclin-like Arabinogalactan Proteins（FLAs）Gene Family in Peach [J]. ACTA HORTICULTURAE SINICA, 2017, 44(4): 653-663.
[6]	WANG Honglei，WANG Yanping，YU Tingqiao，LIU Shengli，CHEN Yuzhen，and LU Cunfu*. Characterization of AmCSDP Gene from Ammopiptanthus mongolicus and the Analysis of Cold Resistance in Transgenic Tobacco Plants [J]. ACTA HORTICULTURAE SINICA, 2017, 44(4): 712-722.
[7]	CAO Ming-ming，YANG Jia，LI Xiao-yu，LI Yu-hua，and LAN Xing-guo. Analysis and Characterization of Interaction Domain Between ARC1 and Exo70A1 from Ornamental Kale（Brassica oleracea var. acephala） [J]. ACTA HORTICULTURAE SINICA, 2015, 42(4): 791-798.
[8]	CHEN Xiu, RAO Xue-Qin, RUAN Xiao-Lei, LIU Fu-Xiu, LI Hua-Ping. Prokaryotic Expression and Antiserum Preparation of Functional Domain of Banana streak virus Coat Protein Gene [J]. ACTA HORTICULTURAE SINICA, 2013, 40(12): 2401-2408.
[9]	LIANG Chun-bo;CHEN Yi-li;LI Ying;and HUANG San-wen;. Overexpression of the Potato Late Blight Resistance Genes RB and R3a and Identification of Domains Required for Recognition Specificity of R3a [J]. ACTA HORTICULTURAE SINICA, 2011, 38(1): 87-94.