Welcome to Acta Agronomica Sinica，

Acta Agronomica Sinica ›› 2018, Vol. 44 ›› Issue (9): 1347-1356.doi: 10.3724/SP.J.1006.2018.01347

• RESEARCH PAPERS • Previous Articles     Next Articles

Cloning and Salt Resistance Function Identification of GmHDL57 Gene from Glycine max

Dan-Xia KE(),Kun-Peng PENG,Meng-Ke ZHANG,Yan JIA,Jing-Jing WANG   

  1. College of Life Sciences / Institute for Conservation and Utilization of Agro-bioresources in Dabie Mountains, Xinyang Normal University, Xinyang 464000, Henan, China
  • Received:2017-11-03 Accepted:2018-06-12 Online:2018-09-10 Published:2018-06-30
  • Contact: Dan-Xia KE E-mail:kdx_029@163.com
  • Supported by:
    This study was supported by the National Natural Science Foundation of China(31400213);Science and Technology Research Projects of Henan Province(182102110448);Funding Scheme for Young Core Teachers of Xinyang Normal University(2015);Nanhu Scholars Program for Young Scholars of Xinyang Normal University, and the Scientific Research Foundation of Graduate School of Xinyang Normal University

RichHTML336

18

PDF

783

PDF

42

Abstract:

The HD-Zip I class transcription factor plays an important role in plant resistance to abiotic stresses. An HD-Zip I class gene GmHDL57 (Glycine max homeodomain-leucine zipper protein 57) was cloned from soybean in this study. Sequence analysis showed that GmHDL57 gene contained a 1038 bp ORF, encoding 345 amino acids, and featured with HD-Zip family proteins’ typical conserved domain. GmHDL57 was expressed in different organs of soybean plants and the highest expression occurred in flowers. The effects of abiotic stresses (abscisic acid, NaCl, PEG, and cold) on GmHDL57 gene expression in soybean seedling stage were analyzed by real-time quantitative PCR. The expression level of GmHDL57 gene was obviously increased under high salinity stress and less affected by ABA and drought stress but decreased by cold stress. The expression level of GmHDL57 gene in roots was significantly higher than that in stems and leaves before and after salt stress, and reached the peak at 48 h, then decreased slowly at 72 h and 96 h after salt stress. The overexpression vector of GmHDL57 was constructed and transformed into Agrobacterium tumefaciens strain EHA105 to obtain the stable transgenic Lotus japonicus plants. After being treated with 200 mmol L -1 NaCl for 14 d, the shoot height, root length, chlorophyll content, root activity as well as K + and Ca 2+ content increased significantly in transgenic plants compared with the wild type. The malondialdehyde content, relative membrane permeability and Na +content were obviously reduced in transgenic plants compared with the wild type. It is hypothesized that the GmHDL57 gene participates in the abiotic stress response of soybean, and over-expression of GmHDL57 gene could enhance resistance to saline in Lotus japonicus.

Key words: Glycine max, GmHDL57 gene, abiotic stress, salt resistance

Fig. 1

Predicted spatial structure of GmHDL57"

Fig. 2

Conserved amino acid sequence alignment analysis of GmHDL57 and its homologous proteins in some other plantsThe sequence marked with solid line demonstrates the homeobox domain, and the sequence marked with dotted line demonstrates the homeobox associated leucine zipper domain."

Fig. 3

Phylogenetic tree of GmHDL57 and its homologsThe scale represents genetic similarity, indicating the proximity relationships among species."

Fig. 4

Relative expression levels of GmHDL57 in different tissuesAbscissa represented young leaf, flower, 1 cm pod, pod shell at 10 d, pod shell at 14 d, seed at 10 d, seed at 14 d, seed at 21 d, seed at 25 d, seed at 28 d, seed at 3 d, seed at 42 d, root and nodule. DAF means days after flower."

Fig. 5

Expression of GmHDL57 treated with various abiotic stresses"

Fig. 6

Expression of GmHDL57 gene in different tissues of soybean under salt stress"

Fig. 7

Positive plants detection and salt stress phenotype analysis of transgenic Lotus japonicusA: detection of GUS gene expression in plants by PCR; B: detection of GmHDL57 gene expression in plants by RT-PCR; C: growth state of Lotus japonicus in different salt concentration treatment for 14 d. Lj 9-5, Lj 1-1: transgenic lines. M: DL2000 DNA marker."

Fig. 8

Changes of shoot height (A) and root length (B) of transgenic Lotus japonicus under salt stressLj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05) and ** means extremely significant difference (P<0.01)."

Fig. 9

Physiological characteristics of transgenic Lotus japonicus under salt stressA: MDA content; B: relative membrane permeability; C: chlorophyll content; D: root activity. Lj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05), ** means extremely significant difference (P<0.01)."

Fig. 10

Cation content in transgenic Lotus japonicus under salt stressA: Na+ content in leaves; B: Na + content in roots; C: K + content in leaves; D: K + content in roots; E: Ca2+ content in leaves; F: Ca2+ content in roots. Lj 9-5, Lj 1-1: transgenic lines. * means significant difference (P<0.05) and ** means extremely significant difference (P<0.01)."

[1] Ariel F D, Manavella P A, Dezar C A, Chan R L . The true story of the HD-Zip family. Trends Plant Sci, 2007,12:419-426
[2] Mukherjee K, Brocchieri L, Burglin T R . A comprehensive classification and evolutionary analysis of plant homeobox genes. Mol Biol Evol, 2009,26:2775-2794
doi: 10.1093/molbev/msp201 pmid: 2775110
[3] Harris J C, Hrmova M, Lopato S, Langridge P . Modulation of plant growth by HD-Zip class I and II transcription factors in response to environmental stimuli. New Phytol, 2011,190:823-837
[4] Henriksson E ,Olsson A S B, Johannesson H, Johansson H, Hanson J, Engström P, Söderman E. , Homeodomain leucine zipper class I genes in Arabidopsis expression patterns and phylogenetic relationships. Plant Physiol, 2005,139:509-518
[5] Olsson A S B, Engstrom P, Soderman E . The homeobox genes ATHB12 and ATHB7 encode potential regulators of growth in response to water deficit in Arabidopsis. Plant Mol Biol, 2004,55:663-677
[6] Himmelbach A, Hoffmann T, Leube M, Höhener B, Grill E . Homeodomain protein ATHB6 is a target of the protein phosphatase ABI1 and regulates hormone responses in Arabidopsis. EMBO J, 2002,21:3029-3038
[7] Ré D A, Dezar C A, Chan R L, Baldwin I T, Bonaventure G . Nicotiana attenuata NaHD20 plays a role in leaf ABA accumulation during water stress, benzylacetone emission from flowers, and the timing of bolting and flower transitions. J Exp Bot, 2011,62:155-166
[8] Ariel F, Diet A, Verdenaud M, Gruber V, Frugier F, Chan R, Crespi M . Environmental regulation of lateral root emergence in Medicago truncatula requires the HD-Zip I transcription factor HB1. Plant Cell, 2010,22:2171-2183
[9] 李明娜, 龙瑞才, 杨青川, 沈益新, 康俊梅, 张铁军 . 紫花苜蓿盐诱导HD-Zip类转录因子MsHB2的克隆及功能分析. 中国农业科学, 2014,47:622-632
Li M N, Long R C, Yang Q C, Shen Y X, Kang J M, Zhang T J . Cloning and function analysis of a salt-stress-induced HD-Zip transcription factor MsHB2 from alfalfa.. Sci Agric Sin, 2014,47:622-632 (in Chinese with English abstract)
[10] Cao L, Yu Y, Duanmu H Z, Chen C, Duan X B, Zhu P H, Chen R R, Li Q, Zhu Y M, Ding X D . A novel Glycine soja homeodomain-leucine zipper (HD-Zip) I gene, Gshdz4, positively regulates bicarbonate tolerance and responds to osmotic stress in Arabidopsis. BMC Plant Biol, 2016,16:184
[11] Chen X, Chen Z, Zhao H, Zhao Y, Cheng B, Xiang Y . Genome-wide analysis of soybean HD-zip gene family and expression profiling under salinity and drought treatments. PLoS One, 2014,9:e87156
[12] Belamkar V, Weeks N T, Bharti A K, Farmer A D, Graham M A, Cannon S B . Comprehensive characterization and RNA-Seq profiling of the HD-Zip transcription factor family in soybean ( Glycine max) during dehydration and salt stress. BMC Genomics, 2014,15:950
[13] Wang Y J, Li Y D, Luo G Z, Tian A G, Wang H W, Zhang J S, Chen S Y . Cloning and characterization of an HD-Zip I gene GmHZ1 from soybean. Planta, 2005,221:831-843
doi: 10.1007/s00425-005-1496-6
[14] 柯丹霞, 李祥永, 王磊, 程琳, 刘永辉, 李小艳, 王慧芳 . 大豆GmHAT5的克隆及其转基因百脉根的抗盐分析. 中国农业科学, 2017,50:1559-1570
Ke D X, Li X Y, Wang L, Cheng L, Liu Y H, Li X Y, Wang H F . Isolation of GmHAT5 from Glycine max and analysis of saline tolerance for transgenic Lotus japonicus.. Sci Agric Sin, 2017,50:1559-1570 (in Chinese with English abstract)
[15] 柯丹霞, 李祥永 . 结瘤信号途径中相关调控蛋白的研究进展. 信阳师范学院学报(自然科学版), 2015,28:621-626
Ke D X, Li X Y . Research progress of key regulatory proteins in nodulation pathway. J Xinyang Nor Univ (Nat Sci Edn), 2015,28:621-626 (in Chinese with English abstract)
[16] Marquez A J . Lotus japonicus Handbook. The Netherlands: Springer, 2005. pp 279-284
[17] Agalou A, Purwantomo S, Overnas E, Johannesson H, Zhu X Y ,Estiati A, de Kam R J, Engström P, Slamet-Loedin I H, Zhu Z, Wang M, Xiong L Z, Meijer A H, Ouwerkerk P B. , A genome-wide survey of HD-Zip genes in rice and analysis of drought-responsive family members. Plant Mol Biol, 2008,66:87-103
[18] Zhang S X, Haider I, Kohlen W, Jiang L, Bouwmeester H, Meijer A H, Schluepmann H, Liu C M, Ouwerkerk P B . Function of the HD-Zip I gene Oshox22 in ABA-mediated drought and salt tolerances in rice. Plant Mol Biol, 2012,80:571-585
doi: 10.1007/s11103-012-9967-1 pmid: 23109182
[19] Zhao Y, Zhou Y Q, Jiang H Y, Li X Y, Gan D F, Peng X J, Zhu S W, Cheng B J . Systematic analysis of sequences and expression patterns of drought-responsive members of the HD-Zip gene family in maize. PLoS One, 2011,6:e28488
[20] Hu R B, Chi X Y, Chai G H, Kong Y Z, He G, Wang X Y, Shi D C, Zhang D Y, Zhou G K . Genome-wide identification, evolutionary expansion, and expression profile of homeodomain- leucine zipper gene family in poplar (Populus trichocarpa). PLoS One, 2012,7:e31149
[21] Liu W, Fu R, Li Q, Li J, Wang L N, Ren Z H . Genome-wide identification and expression profile of homeodomain-leucine zipper class I gene family in Cucumis sativus. Gene, 2013,531:279-287
[22] Fu R, Liu W, Li Q, Li J, Wang L N, Ren Z H . Comprehensive analysis of the homeodomain-leucine zipper IV transcription factor family in Cucumis sativus. Genome, 2013,56:395-405
[23] Elhiti M, Stasolla C . Structure and function of homeodomain-leucine zipper (HD-Zip) proteins. Plant Signal Behav, 2009,4:86-88
doi: 10.4161/psb.4.2.7692 pmid: 2637487
[24] Sahu B B, Shaw B P . Isolation, identification and expression analysis of salt-induced genes in Suaeda maritime, a natural halophyte, using PCR-based suppression subtractive hybridization. BMC Plant Biol, 2009,9:69
[25] 王臻昱, 才华, 柏锡, 纪巍, 李勇, 魏正巍, 朱延明 . 野生大豆GsGST19基因的克隆及其转基因苜蓿的耐盐碱性分析. 作物学报, 2013,38:971-979
Wang Z Y, Cai H, Bai X, Ji W, Li Y, Wei Z W, Zhu Y M . Isolation of GsGST19 from Glycine soja and analysis of saline- alkaline tolerance for transgenic Medicago sativa.. Acta Agron Sin, 2013,38:971-979 (in Chinese with English abstract)
[26] 魏正巍, 朱延明, 化烨, 才华, 纪巍, 柏锡, 王臻昱, 文益东 . 转GsPPCK1基因苜蓿植株的获得及其耐碱性分析. 作物学报, 2013,39:68-75
Wei Z W, Zhu Y M, Hua Y, Cai H, Ji W, Bai X, Wang Z Y, Wen Y D . Transgenic alfalfa with GsPPCK1 and its alkaline tolerance analysis.. Acta Agron Sin, 2013,39:68-75 (in Chinese with English abstract)
[27] 赵阳, 朱延明, 柏锡, 纪巍, 吴婧, 唐立郦, 才华 . 转GsCBRLK/SCMRP双价基因苜蓿耐碱性及氨基酸含量分析. 作物学报, 2014,40:431-438
Zhao Y, Zhu Y M, Bai X, Ji W, Wu J, Tang L L, Cai H . Over-expressing GsCBRLK/SCMRP enhances alkaline tolerance and methionine content in transgenic Medicago sativa. . Acta Agron Sin, 2014,40:431-438 (in Chinese with English abstract)
[28] Yang T, Poovaiah B W . Hydrogen peroxide homeostasis: activation of plant catalase by calcium/calmodulin. Proc Nat Acad Sci USA, 2002,99:4097-4102
doi: 10.1073/pnas.052564899 pmid: 122654
[29] 朱娉慧, 陈冉冉, 于洋, 宋雪薇, 李慧卿, 杜建英, 李强, 丁晓东, 朱延明 . 碱胁迫相关基因GsWRKY15的克隆及其转基因苜蓿的耐碱性分析. 作物学报, 2017,43:1319-1327
Zhu P H, Chen R R, Yu Y, Song X W, Li H Q, Du J Y, Li Q, Ding X D, Zhu Y M . Cloning of gene GsWRKY15 related to alkaline stress and alkaline tolerance of transgenic plants.. Acta Agron Sin, 2017,43:1319-1327 (in Chinese with English abstract)
[30] Olhoft P M, Flagel L E, Donovan C M . Efficient soybean transformation using hygromycin B selection in the cotyledonary-node method. Planta, 2003,216:723-735
doi: 10.1007/s00425-002-0922-2 pmid: 12624759
[31] 王昌陵, 赵军, 李英慧, 范云六, 张丽娟, 刘章雄, 关荣霞, 吕淑霞, 常汝镇, 邱丽娟 . 转录因子ABP9 转化大豆(Glycine max L.)及遗传转化条件优化. 中国农业科学, 2008,41:1908-1916
Wang C L, Zhao J, Li Y H, Fan Y L, Zhang L J, Liu Z X, Guan R X, Lyu S X, Chang R Z, Qiu L J . Transforming transcription factor ABP9 into Soybean and optimization of the transformation system. Sci Agric Sin, 2008,41:1908-1916 (in Chinese with English abstract)
[32] Devi M K, Sakthivela G, Giridhar P . Protocol for augmented shoot organogenesis in selected variety of soybean. J Exp Biol, 2012,50:729-734
[33] 杨权, 王月月, 刘炎光, 蒋春志, 张孟臣, 张洪霞, 张洁, 王冬梅 . 大豆子叶节遗传转化体系优化及抗逆基因AtNHX5的转化研究. 大豆科学, 2015,34:205-211
Yang Q, Wang Y Y, Liu Y G, Jiang C Z, Zhang M C, Zhang H X, Zhang J, Wang D M . Study on optimization of soybean cotyledonary node genetic transformation system and the transformation of resistance gene AtNHX5.. Soybean Sci, 2015,34:205-211 (in Chinese with English abstract)
[34] 柯丹霞, 熊文真, 彭昆鹏, 李祥永 . 抗盐基因Gm01g04890大豆子叶节遗传转化研究. 信阳师范学院学报(自然科学版), 2017,30(1):46-51
Ke D X, Xiong W Z, Peng K P, Li X Y . Study on genetic transformation of salt resistant gene Gm01g04890 in soybean. J Xinyang Nor Univ(Nat Sci Edn), 2017,30(1):46-51 (in Chinese with English abstract)
[1] DONG Yan-Kun, HUANG Ding-Quan, GAO Zhen, CHEN Xu. Identification, expression profile of soybean PIN-Like (PILS) gene family and its function in symbiotic nitrogen fixation in root nodules [J]. Acta Agronomica Sinica, 2022, 48(2): 353-366.
[2] ZHANG Xue-Cui, SUN Su-Li, LU Wei-Guo, LI Hai-Chao, JIA Yan-Yan, DUAN Can-Xing, ZHU Zhen-Dong. Identification of resistance gene against phytophthora root rot in new soybean lines breeded in Henan province [J]. Acta Agronomica Sinica, 2021, 47(2): 275-284.
[3] ZHANG Xue-Cui,ZHONG Chao,DUAN Can-Xing,SUN Su-Li,ZHU Zhen-Dong. Fine mapping of Phytophthora resistance gene RpsZheng in soybean cultivar Zheng 97196 [J]. Acta Agronomica Sinica, 2020, 46(7): 997-1005.
[4] Dai-Ling LIU,Jun-Feng XIE,Qian-Rui HE,Si-Wei CHEN,Yue HU,Jia ZHOU,Yue-Hui SHE,Wei-Guo LIU,Wen-Yu YANG,Xiao-Ling WU. QTL analysis for relative contents of glycinin and β-conglycinin fractions from storage protein in soybean seeds under monoculture and relay intercropping [J]. Acta Agronomica Sinica, 2020, 46(3): 341-353.
[5] CHEN Ying,ZHANG Sheng-Rui,WANG Lan,WANG Lian-Zheng,LI Bin,SUN Jun-Ming. Characteristics of oil components and its relationship with domestication of oil components in wild and cultivated soybean accessions [J]. Acta Agronomica Sinica, 2019, 45(7): 1038-1049.
[6] Dan-Xia KE,Kun-Peng PENG,Yan JIA,Shuo ZENG,Ying-Zhi WANG,Jing-Yi ZHANG. Functional Characterization of Soybean Cystatins Gene GmCYS2 [J]. Acta Agronomica Sinica, 2018, 44(8): 1159-1168.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
Add: No. 80 Xueyuan South Rd, Beijing 100081, P. R. China E-mail: zwxb301@caas.cn
Supported by Beijing Magtech Co.Ltd