Whole Genome Identification of Barley NRAMP and Gene Expression Analysis Under Heavy Metal Stress

doi:10.13560/j.cnki.biotech.bull.1985.2021-1006

Abstract

Abstract:

Natural resistance associated macrophage protein（NRAMP）is an important type of membrane transporter in plants,it plays an important role in transporting and reusing heavy metal ions. Studying the mechanism of NRAMP family gene expression regulation under Mn²⁺,Cd²⁺ and Cu²⁺ stress would provide a theoretical basis for further revealing the roles of the NRAMP family genes against heavy metal stress. Bioinformatics methods were used to screen and identify NRAMP family gene sequences from the whole genome of Hordeum vulgare L.,analyzes the gene structure,conserved motifs,evolutionary characteristics and expression profiles of its family members. Moreover,qRT-PCR method was applied to detect the expression of the NRAMP gene in barley seedlings under different heavy metal ion stress. The results showed that there were 10 NRAMP family members in the barley genome,which were mainly located on the cell membrane. Phylogenetic analysis demonstrated that the conserved motifs and gene structures of closely related family members were similar,and the genetic relationship between barley and rice NRAMP family members was closer. The results of quantitative analysis showed that the expression levels of HvNramp1 and HvNramp2 were significantly up-regulated under Mn²⁺ stress. Under Cd²⁺stress,all genes’ expression levels were up-regulated except HvNramp8. Under Cu²⁺ stress,the expression levels of the remaining 6 genes except HvNramp3 were significantly up-regulated. The above results indicate that the NRAMP family genes can respond to the stress of heavy metal ions through positive regulation.

Key words: barley, NRAMP gene family, heavy metal stress, physiological indexes, gene expression

LI Yi-han, YU Lang-liu, LI Chun-yan, ZHANG Meng-meng, ZHANG Xiao-qin, FANG Yun-xia, XUE Da-wei. Whole Genome Identification of Barley NRAMP and Gene Expression Analysis Under Heavy Metal Stress[J]. Biotechnology Bulletin, 2022, 38(6): 103-111.

Figures/Tables 7

References 31

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基因 Gene	基因ID Gene ID	引物 Primer	正向引物序列 Forward primer sequence（5'-3'）	反向引物序列 Reverse primer sequence（5'-3'）
HvNramp1	HORVU3Hr1G042440	HvqRT1	CTACCTTCGTGAGTTGGCTTGTA	TGTTGTCTTGTATTGGCACCTCT
HvNramp2	HORVU4Hr1G013220	HvqRT2	AGCACCATAACCGGGACCTA	TGAGCCACTCGTTGAGGACAT
HvNramp3	HORVU4Hr1G069320	HvqRT3	TGTTTACCTCGCCTTCATCGT	CTACCCACTTCCGCTCATCTT
HvNramp4	HORVU4Hr1G077510	HvqRT4	AGGACCTCGCAGACATACCG	AGAAGGACGGATAAAATGCTACG
HvNramp5	HORVU4Hr1G090460	HvqRT5	GGTCTCCAAGCTGGGATGTCT	ATGGCTGTGATAAAATCATCCTG
HvNramp6	HORVU5Hr1G018850	HvqRT6	AACCGAGCCGTACAATGGAC	AGATGATCGTGGCGATACGC
HvNramp7	HORVU5Hr1G050310	HvqRT7	CTGCATAGCAGGTCTCGCACT	CATTTCCACCCAGAAGTGCC
HvNramp8	HORVU7Hr1G078330	HvqRT8	GTGAGGTGAACACTGGCCAGA	CCACCATTATGCCTAAAGAGCC
HvNramp9	HORVU5Hr1G050330	HvqRT9	ATCGGGATAAACGTCTACTTCCT	ATGACAGAGGCGACGTAGAGC
HvNramp10	HORVU7Hr1G106570	HvqRT10	GTGCTAACATGGGCGATCG	ACGGCAAGGTACACTTCCTGTT
HvActin	HORVU1Hr1G002840	Actin	TGGATCGGAGGGTCCATCCT	GCACTTCCTGTGGACGATCGCTG

基因 Gene	基因ID Gene ID	引物 Primer	正向引物序列 Forward primer sequence（5'-3'）	反向引物序列 Reverse primer sequence（5'-3'）
HvNramp1	HORVU3Hr1G042440	HvqRT1	CTACCTTCGTGAGTTGGCTTGTA	TGTTGTCTTGTATTGGCACCTCT
HvNramp2	HORVU4Hr1G013220	HvqRT2	AGCACCATAACCGGGACCTA	TGAGCCACTCGTTGAGGACAT
HvNramp3	HORVU4Hr1G069320	HvqRT3	TGTTTACCTCGCCTTCATCGT	CTACCCACTTCCGCTCATCTT
HvNramp4	HORVU4Hr1G077510	HvqRT4	AGGACCTCGCAGACATACCG	AGAAGGACGGATAAAATGCTACG
HvNramp5	HORVU4Hr1G090460	HvqRT5	GGTCTCCAAGCTGGGATGTCT	ATGGCTGTGATAAAATCATCCTG
HvNramp6	HORVU5Hr1G018850	HvqRT6	AACCGAGCCGTACAATGGAC	AGATGATCGTGGCGATACGC
HvNramp7	HORVU5Hr1G050310	HvqRT7	CTGCATAGCAGGTCTCGCACT	CATTTCCACCCAGAAGTGCC
HvNramp8	HORVU7Hr1G078330	HvqRT8	GTGAGGTGAACACTGGCCAGA	CCACCATTATGCCTAAAGAGCC
HvNramp9	HORVU5Hr1G050330	HvqRT9	ATCGGGATAAACGTCTACTTCCT	ATGACAGAGGCGACGTAGAGC
HvNramp10	HORVU7Hr1G106570	HvqRT10	GTGCTAACATGGGCGATCG	ACGGCAAGGTACACTTCCTGTT
HvActin	HORVU1Hr1G002840	Actin	TGGATCGGAGGGTCCATCCT	GCACTTCCTGTGGACGATCGCTG

基因 Gene	蛋白质 Protein	氨基酸长度 Amino acid length/aa	分子质量 Molecular mass/Da	理论等电点 Theoretical isoelectric point	亚细胞定位 Subcellular localization	跨膜结构域 Transmembrane domain	染色体定位 Chromosomal localization	物理图谱位置 Physical map location/Mb
HvNramp1	A0A287KVZ6	606	66 306.78	7.82	细胞膜Cell membrane	12	Chr.3H	256.8
HvNramp2	A0A287N9P9	549	60 120.01	5.07	细胞膜Cell membrane	10	Chr.4H	43.2
HvNramp3	M0XUV1	522	56 885.5	5.76	细胞膜Cell membrane	10	Chr.4H	543.5
HvNramp4	M0X4N6	542	58 612.97	7.03	细胞膜Cell membrane	11	Chr.4H	576.7
HvNramp5	A0A287Q4H3	212	22 845.85	5.17	细胞膜Cell membrane	5	Chr.4H	615.3
HvNramp6	F2EHM2	547	58 995.65	5.2	细胞膜Cell membrane	11	Chr.5H	75.1
HvNramp7	A0A287R642	248	58 995.65	5.41	细胞膜Cell membrane	7	Chr.5H	373.2
HvNramp8	A0A287X031	515	55 875.86	8.12	细胞核Nucleus	5	Chr.7H	438.5
HvNramp9	A0A287R6C2	1 010	110 400.77	6.13	细胞膜Cell membrane	11	Chr.5H	373.2
HvNramp10	M0W7C8	548	59 571.99	7.99	细胞膜Cell membrane	12	Chr.7H	591.4

基因 Gene	蛋白质 Protein	氨基酸长度 Amino acid length/aa	分子质量 Molecular mass/Da	理论等电点 Theoretical isoelectric point	亚细胞定位 Subcellular localization	跨膜结构域 Transmembrane domain	染色体定位 Chromosomal localization	物理图谱位置 Physical map location/Mb
HvNramp1	A0A287KVZ6	606	66 306.78	7.82	细胞膜Cell membrane	12	Chr.3H	256.8
HvNramp2	A0A287N9P9	549	60 120.01	5.07	细胞膜Cell membrane	10	Chr.4H	43.2
HvNramp3	M0XUV1	522	56 885.5	5.76	细胞膜Cell membrane	10	Chr.4H	543.5
HvNramp4	M0X4N6	542	58 612.97	7.03	细胞膜Cell membrane	11	Chr.4H	576.7
HvNramp5	A0A287Q4H3	212	22 845.85	5.17	细胞膜Cell membrane	5	Chr.4H	615.3
HvNramp6	F2EHM2	547	58 995.65	5.2	细胞膜Cell membrane	11	Chr.5H	75.1
HvNramp7	A0A287R642	248	58 995.65	5.41	细胞膜Cell membrane	7	Chr.5H	373.2
HvNramp8	A0A287X031	515	55 875.86	8.12	细胞核Nucleus	5	Chr.7H	438.5
HvNramp9	A0A287R6C2	1 010	110 400.77	6.13	细胞膜Cell membrane	11	Chr.5H	373.2
HvNramp10	M0W7C8	548	59 571.99	7.99	细胞膜Cell membrane	12	Chr.7H	591.4