Cloning and Expression of ZmXTH23 in Maize (Zea mays) and Its Response to Salt and Drought Stress
CHEN Dong-Bin1, 3, WANG Qian-Qian2, 3, SUN Zhi-Yi2, 3, YANG Xiao-Ying2, 3, FU Jing-Xiao2, 3, GUO Xin-Mei2, 3, *, SONG Xi-Yun2, 3, *
1 College of Life Science, Qingdao Agricultural University, Qingdao 266109, China; 2 College of Agronomy, Qingdao Agricultural University, Qingdao 266109, China; 3 Key Laboratory of Major Crop Germplasm Innovation and Application in Qingdao, Qingdao 266109, China
Abstract：Xyloglucan transglycosidase/hydrolase (XTH) is a key enzyme in the process of plant cell wall remodeling, and participates in the regulation of plant growth and development. To elucidate and explore the function of XTH in the process of stress response in maize (Zea mays), ZmXTH23 (GenBank No. LOC100191584) in XTH family was cloned from maize inbred line 'Chang7-2' and its biological function was studied. Bioinformatics analysis showed that ZmXTH23 contained a complete open reading frame of 897 bp encoded 298 amino acids and was a member of LamG superfamily. Besides, it performed xyloglucan endotransglycosidase (XET) activity and its amino acid sequence had the closest affinity with that of PmXTH25 (GenBank No. RLM56175.1) in Panicum miliaceum (similarity reached 90%). The expression pattern of ZmXTH23 was analyzed by qRT-PCR, and the results showed that the expression of ZmXTH23 was tissue-specific, it had the highest expression level in young stem (P＜0.01), and was induced by abscisic acid (ABA), NaCl and PEG6000. Prokaryotic expression analysis by SDS-PAGE and Western blot confirmed that ZmXTH23 protein could be expressed in Escherichia coli BL21 transferred with pET28a-ZmXTH23 recombinant plasmid. Besides, the growth of host bacteria pET28a-ZmXTH23 under different concentrations of salt and mannitol stress was better than that of host bacteria pET28a, which indicated that the ZmXTH23 protein enhanced the salt tolerance and drought resistance of pET28a-ZmXTH23 recombinant host strain. In conclusion, ZmXTH23 played an important role in response to abiotic stress and the results in this study provides theoretical basis for the creation of new maize germplasm resistant to stress.
陈东滨, 王茜茜, 孙智仪, 杨小英, 傅经效, 郭新梅, 宋希云. 玉米ZmXTH23的克隆、表达及其对盐胁迫和干旱胁迫的响应[J]. 农业生物技术学报, 2019, 27(9): 1533-1541.
CHEN Dong-Bin, WANG Qian-Qian, SUN Zhi-Yi, YANG Xiao-Ying, FU Jing-Xiao, GUO Xin-Mei, SONG Xi-Yun. Cloning and Expression of ZmXTH23 in Maize (Zea mays) and Its Response to Salt and Drought Stress. 农业生物技术学报, 2019, 27(9): 1533-1541.
1 韩彦莎, 仪慧兰. 2016. 过表达胡杨XTH基因能够提高烟草抗旱性[J]. 中国生物化学与分子生物学报, 32(8): 919-925. (Han Y S, Yi H L.2016. Over-expression of Populus euphratica XTH gene enhances drought tolerance of tobacco[J]. Chinese Journal of Biochemistry and Molecular Biology, 32(8): 919-925) 2 贾新平, 衍明, 孙晓波, 等. 2015. 胁迫对海滨雀稗生长和生理特性的影响[J]. 草业学报, 24(12): 204-212. (Jia X P, Deng Y M, Sun X B.2015. Impacts of salt stress on the growth and physiological characteristics of Paspalum vaginatu[J]. Journal of Grass Industry, 24(12): 204-212.) 3 韦存虚, 王建波, 陈义芳, 等. 2004. 盐生植物星星草叶表皮具有泌盐功能的蜡质层[J]. 生态学报, 24(11): 2451-2456. (Wei C X, Wang J B, Chen Y F, et al.2004. Epicuticular wax of leaf epidermis: A functional structure for salt excretion in ahalophyte Puccinellia tenuiflora[J]. Acta Ecologica Sinica, 24(11): 2451-2456.) 4 徐丽华, 刘春雷, 常玉梅, 等. 2011. 双标准曲线相对定量PCR试验原理与方法[J]. 生物技术通报, 2011(1): 70-75. (Xu L H, Liu C L, Chang Y M, et al.2011. Theory and method of double-standard curves method of relative quantification PCR[J]. Biotechnology Bulletin, 2011(1): 70-75.) 5 Amor Y, Haigler C H, Johnson S, et al.1995. A membrane-associated form of sucrose synthase and its potential role in synthesis of cellulose and callose in plants[J]. Proceedings of the National Academy of Sciences of the USA, 92(20): 9353-9357. 6 Aranjuelo I, Molero G, Erice G, et al.2011. Plant physiology and proteomics reveals the leaf response to drought in alfalfa (Medicago sativa L.)[J]. Journal of Experimental Botany, 62(1): 111-123. 7 Ashoub A, Beckhaus T, Berberich T.2013. Comparative analysis of barley leaf proteome as affected by drought stress[J]. Planta, 237(3): 771-781. 8 Benesova M, Hola D, Fischer L, et al.2012. The physiology and proteomics of drought tolerance in maize: Early stomatal closure as a cause of lower tolerance to short-term dehydration?[J]. PLOS ONE, 7(6): e38017. 9 Blumwald E.2000. Sodium transport and salt tolerance in plants[J]. Current Opinion in Cell Biology, 12(4): 431-434. 10 Chen J H, Jiang H W, Hsieh E J, et al.2012. Drought and salt stress tolerance of an Arabidopsis glutathione S-Transferase U17 knockout mutant are attributed to the combined effect of glutathione and abscisic acid[J]. Plant Physiology, 158(1): 340-351. 11 Cho S K, Kim J E, Park J A, et al.2006. Constitutive expression of abiotic stress-inducible hot pepper CaXTH3, which encodes a xyloglucan endotransglucosylase/hydrolase homolog, improves drought and salt tolerance in transgenic arabidopsis plants[J]. The Federation of European Biochemical Societies Letters, 580(13): 3136-3144. 12 Choi J Y, Seo Y S, Kim S J, et al.2011. Constitutive expression of CaXTH3, a hot pepper xyloglucan endotransglucosylase/hydrolase, enhanced tolerance to salt and drought stresses without phenotypic defects in tomato plants (Solanum lycopersicum cv. Dotaerang)[J]. Plant Cell Reports, 30(5): 867-877. 13 Dong J L, Jiang Y Y, Chen R J, et al.2011. Isolation of a novel xyloglucan endotransglucosylase (OsXET9) gene from rice and analysis of the response of this gene to abiotic stresses[J]. African Journal of Biotechnology, 10(76): 17424-17434. 14 Fry S C, Smith R C, Renwick K F, et al.1992. Xyloglucan endotransglycosylase, a new wall-loosening enzyme activity from plants[J]. Biochemical Journal, 282(Pt3): 821-828. 15 Harada T, Torii Y, Morita S, et al.2011. Cloning, characterization and expression of xyloglucan endotransglucosylase/hydrolase and expansin genes associated with petal growth and development during carnation flower opening[J]. Journal of Experimental Botany, 62(2): 815-823. 16 He H, Serraj R, Yang Q.2009. Changes in OsXTH gene expression, ABA content, and peduncle elongation in rice subjected to drought at the reproductive stage[J]. Acta Physiology Plant, 31(4): 749-756. 17 Hu J, Ma Q, Kuma R T, et al.2016. ZxSKOR is important for salinity and drought tolerance of Zygophyllum xanthoxylum by maintaining K+ homeostasis[J]. Plant Growth Regulation, 80(2): 195-205. 18 Hu X L, Wu L J, Zhao F Y, et al.2015. Phosphoproteomic analysis of the response of maize leaves to drought, heat and their combination stress[J]. Frontiers in Plant Science, 6: 298. 19 Lee B R, Kim K Y, Jung W J, et al.2007. Peroxidases and lignification in relation to the intensity of water-deficit stress in white clover (Trifolium repens L.)[J]. Journal of Experimental Botany, 58(6): 1271-1279. 20 Matsui A, Yokoyama R, Seki M, et al.2005. AtXTH27 plays an essential role in cell wall modification during the development of tracheary elements[J]. The Plant Journal, 42(4): 525-534. 21 Miedes E, Lorences E P.2009. Xyloglucan endotransglucosylase/hydrolases (XTHs) during tomato fruit growth and ripening[J]. Journal of Plant Physiology, 166(5): 489-498. 22 Miedes E, Suslov D, Vandenbussche F, et al.2013. Xyloglucan endotransglucosylase/hydrolase (XTH) overexpression affects growth and cell wall mechanics in etiolated Arabidopsis hypocotyls[J]. Journal of Experimental Botany, 64(8): 2481-2497. 23 Miedes E, Zarra I, Hoson T, et al.2011. Xyloglucan endotransglucosylase and cell wall extensibility[J]. Journal of Plant Physiology, 168(3): 196-203. 24 Miller G, Suzuki N, Ciftciyilmaz S, et al.2010. Reactive oxygen species homeostasis and signalling during drought and salinity stresses[J]. Plant Cell and Environment, 33(4): 453-467. 25 Mohammadi P P, Moieni A, Hiraga S, et al.2012. Organ-specific proteomic analysis of drought-stressed soybean seedlings[J]. Journal of Proteomics, 75(6): 1906-1923. 26 Nishitani K, Tominaga R.1992. Endo-xyloglucan transferase, a novel class of glycosyltransferase that catalyzes transfer of a segment of xyloglucan molecule to another xyloglucan molecule[J]. Journal of Biological Chemistry, 267(29): 21058-21064. 27 Nishikubo N, Takahashi J, Roos A A, et al.2011. Xyloglucan endotransglycosylase-mediated xyloglucan rearrangements in developing wood of hybrid aspen[J]. Plant Physiology, 155(1): 399-413. 28 Nishiyama R, Le D T, Watanabe Y, et al.2012. Transcriptome analyses of a salt-tolerant cytokinin-deficient mutant reveal differential regulation of salt stress response by cytokinin deficiency[J]. PLOS ONE, 7(2): e32124. 29 Osato Y, Yokoyama R, Nishitani K.2006. A principal role for AtXTH18 in Arabidopsis thaliana root growth: A functional analysis using RNAi plants[J]. Journal of Plant Research, 119(2): 153-162. 30 Osmond C B, Grace S C.1995. Perspectives on photoinhibition and photorespiration in the field: Quintessentialin efficiencies of the light and dark reactions of photosynthesis?[J]. Journal of Experimental Botany, 46: 1351-1362. 31 Raorane M L, Pabuayon I M, Varadarajan A R.2015. Proteomic insights into the role of the large-effect QTL q DTY12.1 for rice yield under drought[J]. Molecular Breeding, 35(6): 139. 32 Rose J K, Braam J, Fry S C, et al.2002. The XTH family of enzymes involved in xyloglucan endotransglucosylation and endohydrolysis: Current perspectives and a new unifying nomenclature[J]. Plant and Cell Physiology, 43(12): 1421-1435. 33 Saladié M, Rose J K, Cosgrove D J, et al.2006. Characterization of a new xyloglucan endotransglucosylase/hydrolase (XTH) from ripening tomato fruit and implications for the diverse modes of enzymic action[J]. The Plant Journal, 47(2): 282-295. 34 Singh A P, Tripathi S K, Nath P, et al.2011. Petal abscission in rose is associated with the differential expression of two ethylene-responsive xyloglucan endotransglucosylase/hydrolase genes, RbXTH1 and RbXTH2[J]. Journal of Experimental Botany, 62(14): 5091-5103. 35 Skirycz A, Inze D.2010. More from less: Plant growth under limited water[J]. Current Opinion in Biotechnology, 21(2): 197-203. 36 Song L, Prince S, Valliyodan B, et al.2016. Genome-wide transcriptome analysis of soybean primary root under varying water-deficit conditions[J]. BMC Genomics, 17(1): 57. 37 Vissenberg K, Fry S C, Pauly M, et al.2005. XTH acts at the microfibril-matrix interface during cell elongation[J]. Journal of Experimental Botany, 56(412): 673-683. 38 Wei C, Cui Q, Zhang X Q, et al.2016. Three P5CS genes including a novel one from Lilium regale play distinct roles in osmotic, drought and salt stress tolerance[J]. Journal of Plant Biology, 59(5): 456-466. 39 Wu Y, Jeong B R, Fry S C, et al.2005. Change in XET activities, cell wall extensibility and hypocotyl elongation of soybean seedlings at low water potential[J]. Planta, 220(4): 593-601. 40 Xue Y F, Liu Z P.2008. Antioxidant enzymes and physiological characteristics in two Jerusalem artichoke cultivars under salt stress[J]. Russian Journal of Plant Physiology, 55(6): 776-781. 41 Zhang Q.2007. Strategies for developing green super rice[J]. Proceedings of the National Academy of Sciences of the USA, 104(42): 16402-16409.
