Cloning of Soybean GmGolS1 and Identification of Heat Resistance in Transgenic Tobacco

LI Ming-yang,QIU Shuang,HE Jia-qi,YU Hai-wei,ZHANG Jun and ZHAI Ying

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Journal of Plant Genetic Resources ›› 2022, Vol. 23 ›› Issue (2) : 575-582. DOI: 10.13430/j.cnki.jpgr.20210823001
Research Articles

Cloning of Soybean GmGolS1 and Identification of Heat Resistance in Transgenic Tobacco

  • LI Ming-yang1, QIU Shuang1, HE Jia-qi1, YU Hai-wei1, ZHANG Jun2, ZHAI Ying1
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Abstract

Raffinose family oligosaccharides(RFOs)are one of important osmoregulatory substance in plants. Galactinol synthase(GolS)is a key enzyme in the synthesis of RFOs. In this study,GmGolS1 was cloned from soybean based on sequence homology. GmGolS1 was found on chromosome 3 with three introns. The full length open reading frame(ORF)of GmGolS1 was 1020 bp,which encodes 339 amino acids with the molecular weight of 38.82 kDa,and the isoelectric point of 5.47. The amino acid sequence GmGolS1 has characteristics that are commonly detected in plant GolS proteins:a conserved serine phosphorylation site,a manganese ion binding associated DXD element,and a carboxy-terminal hydrophobic pentapeptide. The phylogenetic analysis showed that GmGolS1 was closely related to AmGolS and MtGolS. Real-time fluorescence quantitative PCR analysis showed that GmGolS1 was inducible upon heat,low temperature,drought and salt stresses in different degrees in seedlings,and the most abundant expression was observed under heat stress treatment. By transforming GmGolS1 was into tobacco,four transgenic lines were identified using genomic DNA PCR and real-time fluorescence quantitative PCR. Test for GolS enzyme activity showed that the transcribed product of GmGolS1 in transgenic tobacco played the catalytic activity of GolS. Gained from plant phenotype,electrolyte leakage,soluble carbohydrate content and malondialdehyde content of GmGolS1 transgenic tobacco under heat stress,GmGolS1 was found correlating with improved heat resistance in transgenic tobacco.

Key words

soybean;galactinol synthase;abiotic stress;heat resistance

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LI Ming-yang,QIU Shuang,HE Jia-qi,YU Hai-wei,ZHANG Jun and ZHAI Ying. Cloning of Soybean GmGolS1 and Identification of Heat Resistance in Transgenic Tobacco. Journal of Plant Genetic Resources. 2022, 23(2): 575-582 https://doi.org/10.13430/j.cnki.jpgr.20210823001

References

[1] Peters S, Mundree S G, Thomson J A, Farrant J M, Keller F. Protection mechanisms in the resurrection plant Xerophyta viscosa (Baker): both sucrose and raffinose family oligosaccharides (RFOs) accumulate in leaves in response to water deficit. Journal of Experimental Botany, 2007, 58(8): 1947-1956
[2] Lahuta L B, Górecki R J. Raffinose in seedlings of winter vetch (Vicia villosa Roth.) under osmotic stress and followed by recovery. Acta Physlologlae Plantarum, 2011, 33(3): 725-733
[3] Nishizawa A, Yabuta Y, Shigeoka S. Galactinol and raffinose constitute a novel function to protect plants from oxidative damage. Plant Physiology, 2008, 147(3): 1251-1263
[4] Frydman R B, Neufeld E F. Synthesis of galactosylinositol by extracts from peas. Biochemical and Biophysical Research Communications, 1963, 12(2): 121-125
[5] Thomas P, Andreas R. Biochemistry and physiology of raffinose family oligosaccharides and galactosyl cyclitols in seeds. Seed Science Research, 2001, 11(3): 185-197
[6] Selvaraj M G, Ishizaki T, Valencia M, Ogawa S, Dedicova B, Ogata T, Yoshiwara K, Maruyama K, Kusano M, Saito K, Takahashi F, Shinozaki K, Nakashima K, Ishitani M. Overexpression of an Arabidopsis thaliana galactinol synthase gene improves drought tolerance in transgenic rice and increased grain yield in the field. Plant Biotechnology Journal, 2007, 15(11): 1465-1477
[7] Shimosaka E, Ozawa K. Overexpression of cold-inducible wheat galactinol synthase confers tolerance to chilling stress in transgenic rice. Breeding Science, 2015, 65(5): 363-371
[8] Vinson C C, Mota A P, Porto B N, Oliveira T N, Sampaio I, Lacerda A L, Danchin E G J, Guimaraes P M, Williams T C R, Brasileiro A C M. Characterization of raffinose metabolism genes uncovers a wild Arachis galactinol synthase conferring tolerance to abiotic stresses. Scientific Reports, 2020, 10(1): 15258-15258
[9] Panikulangara T J, Eggers S G, Wunderlich M, Stransky H, Scho?ffl F. Galactinol synthase1. A novel heat shock factor target gene responsible for heat-induced synthesis of raffinose family oligosaccharides in Arabidopsis. Plant Physiology, 2004, 136(2): 3148-3158
[10] Pillet J, Egert A, Pieri P, Lecourieux F, Kappel C, Charon J, Gomès E, Keller F, Delrot S, Lecourieux D. VvGOLS1 and VvHsfA2 are involved in the heat stress responses in grapevine berries. Plant and Cell Physiology, 2012, 53(10): 1776-1792
[11] Wang D H, Yao W, Song Y, Liu W C, Wang Z Z. Molecular characterization and expression of three galactinol synthase genes that confer stress tolerance in Salvia miltiorrhiza. Journal of Plant Physiology, 2012, 169(18): 1838-1848
[12] 谷雷. 玉米肌醇半乳糖苷合成酶2基因(ZmGolS2)启动子上响应热激的顺式作用元件鉴定. 西北农林科技大学, 2014
Gu L. Identification of the heat response Cis-acting elements of the ZmGolS2 gene promoter. Northwest A&F University, 2014
[13] Salvi P, Kamble N U, Majee M. Stress-inducible galactinol synthase of chickpea (CaGolS) is implicated in heat and oxidative stress tolerance through reducing stress-induced excessive reactive oxygen species accumulation. Plant Cell Physiology, 2018, 59(1): 155-166
[14] 汪明华, 李佳佳, 陆少奇, 邵文韬, 程安东, 张文明, 王晓波, 邱丽娟. 大豆品种耐高温特性的评价方法及耐高温种质筛选与鉴定. 植物遗传资源学报, 2019, 20(4): 891-902
Wang M H, Li J J, Lu S Q, Shao W T, Cheng A D, Zhang W M, Wang X B, Qiu L J. Construction of evaluation standard for tolerance to high-temperature and screening of heat-tolerant germplasm resources in soybean. Journal of Plant Genetic Resources, 2019, 20(4): 891- 902
[15] Qiu S, Zhang J, He J Q, Sha W, Li M Y, Zhao Y, Zhai Y. Overexpression of GmGolS2-1, a soybean galactinol synthase gene, enhances transgenic tobacco drought tolerance. Plant Cell Tissue and Organ Culture, 2020, 143(3): 507-516
[16] 邱爽, 张军, 何佳琦, 李铭杨, 周雨明, 邬长乐, 袁洪淼, 刘嘉仪, 翟莹. 大豆GmGolS2-1基因高温胁迫诱导表达及转基因烟草鉴定. 江苏农业学报, 2021, 37(1): 38-43
Qiu S, Zhang J, He J Q, Li M Y, Zhou Y M, Wu C L, Yuan H M, Liu J Y, Zhai Y. Expression of soybean GmGolS2-1 induced by heat stress and identification of GmGolS2-1 transgenic tobacco. Jiangsu Journal of Agricultural Science, 2021, 37(1): 38-43
[17] 何佳琦, 翟莹, 张军, 邱爽, 李铭杨, 赵艳, 张梅娟, 马天意. 大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达. 浙江农业学报, 2021, 33(1): 1-7
He J Q, Zhai Y, Zhang J, Qiu S, Li M Y, Zhao Y, Zhang M J, Ma T Y. Cloning and expression analysis of GmDof1.5 in soybean under abiotic stress. Acta Agriculturae Zhejiangensis, 2021, 33(1): 1-7
[18] Hoekema A, Hirsch P R, Hooykaas P J J, Schilperoort R A. A binary plant vector strategy based on separation of vir- and T-region of the Agrobacterium tumefaciens Ti-plasmid. Nature, 1983, 303(5913): 179-180
[19] Zhai Y, Wang Y, Li Y J, Lei T T, Yan F, Su L T, Li X W, Zhao Y, Sun X, Li J W, Wang Q Y. Isolation and molecular characterization of GmERF7, a soybean ethylene-response factor that increases salt stress tolerance in tobacco. Gene, 2013, 513(1): 174-183
[20] Bates L S, Waldren R P, Teare I D. Rapid determination of free proline for water-stress studies. Plant and Soil,1973, 39(1): 205-207
[21] Shao H B, Liang Z S, Shao M A. Osmotic regulation of 10 wheat (Triticum aestivum L.) genotypes at soil water defcits. Colloids Surf B Biointerfaces, 2006, 47(2): 132-139
[22] Zhao C, Liu B, Piao S, Wang X, Lobell D B, Huang Y, Huang M, Yao Y, Bassu S, Ciais P, Durand J L, Elliott J, Ewert F, Janssens I A, Li T, Lin E, Liu Q, Martre P, Müller C, Peng S, Pe?uelas J, Ruane A C, Wallach Da, Wang T, Wu D, Liu Z, Zhu Y, Zhu Z, Asseng S. Temperature increase reduces global yields of major crops in four independent estimates. Proceedings of the National Academy of Sciences of the United States of America, 2017, 114 (35): 9326-9331
[23] dos Santos T B, Budzinski I G, Marur C J, Petkowicz C L, Pereira L F, Vieira L G. Expression of three galactinol synthase isoforms in Coffea arabica L. and accumulation of raffinose and stachyose in response to abiotic stresses. Plant Physiology and Biochemistry, 2011, 49(4): 441-448
[24] Taji, T, Ohsumi, C, Iuchi, S, Seki, M, Kasuga M, Kobayashi M, Yamaguchi-Shinozaki K, Shinozaki, K. Important roles of drought- and cold-inducible genes for galactinol synthase in stress tolerance in Arabidopsis thaliana. The Plant Journal, 2002, 29(4): 417-426
[25] Cakmak I, Horst W J. Effect of aluminium on lipid peroxidation, superoxide dismutase, catalase, and peroxidase activities in root tips of soybean (Glycinemax). Physiologia Plantarum, 1991, 83(3): 463-468

Funding

Training Plan of Young Creative Talents of Average Four-year College of Heilongjiang(UNPYSCT-2017153),InnovativeResearch Project for Graduate Students of Qiqihar University(YJSCX2020041)
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