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Cloning and Drought Resistance Identification of TaXTH-7A Gene in Wheat (Triticum aestivum) |
ZHOU Qi, FENG Yan-Ru, LI Song, LIU Zi-Hui, ZHENG Wei-Jun*, CHAI Shou-Cheng* |
College of Agronomy, Northwest A&F University, Yangling 712100, China |
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Abstract Wheat (Triticum aestivum) is one of the important food crops in China. However, drought and other adverse conditions have seriously affected the quality and yield of wheat.Screening of stress-resistant genes and analyzing their mechanism of action can provide candidate genes and theoretical basis for wheat resistance molecular breeding. Xyloglucan endotransglucosylase/hydrolase (XET/XEH) is a class of enzymes with xyloglucan endoglycosyltransferase /hydrolase activity, which are collectively referred to as XTH, belonging to glycoside hydrolase 16 (GH 16) family. It can participate in the formation and remodeling of cell wall and plays an important role in plant growth and development and stress resistance. In this study, the cDNA sequence of the TaXTH-7A gene (GenBank No. MK395550) was isolated by PCR from wheat, and the coding region was 870 bp in length, which encoded 290 amino acid. The TaXTH-7A genome contained 3 exons and 2 introns. Protein sequence analysis indicated that TaXTH-7A contains several XET-specific domains and they were highly conserved across species. Phylogenetic tree analysis indicated that the XTH-7A protein evolved between monocotyledonous and dicotyledonous, and between C3 and C4 plants, and was closely related to Aegilops tauschii. The results of qRT-PCR showed that TaXTH-7A gene was expressed in roots, stems, leaves and spiket of wheat at heading stage, and it was dominant in roots. After drought stress, its expression increased and peaked at 4 hours. Subcellular localization results showed that TaXTH-7A was localized in the cytoplasm, cell membrane, and apoplast. Functional identification showed that over-expression of tTaXTH-7A enhanced drought resistance of Arabidopsis thaliana. This study is helpful to further study the function of TaXTH-7A gene in wheat, and also provides a theoretical basis for wheat resistance to abiotic stress.
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Received: 06 March 2019
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Corresponding Authors:
zhengweijun@nwsuaf.edu.cn; chaishoucheng@126.com
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1 姜慧芳, 任小平. 2004. 干旱胁迫对花生叶片SOD活性和蛋白质的影响[J]. 作物学报, 30(2): 169-174. (Jang H F, Ren X P.2004. The effect on SOD activity and protein content in groundnut leaves by drought stress[J]. Acta Agronomica Sinica, 3(1): 1-4.) 2 黎裕, 王建康, 邱丽娟, 等. 2010. 中国作物分子育种现状与发展前景[J]. 作物学报, 36(9): 1425-1430. (Li Y, Wang J K, Qiu L J, et al.2010. Crop molecular breeding in china: current status and perspectives[J]. Acta Agronomica Sinica, 36(09): 1425-1430.) 3 李莉, 田士林. 2007. 不同小麦材料抗旱性比较[J]. 安徽农业科学, 35(24): 7409-7409. (Li L, Tian S L.2007. Comparison of drought-resistance among difference wheat materials[J]. Journal of Anhui Agricultural Sciences, 35(24): 7409-7409.) 4 李嵩, 李敏, 刘子辉, 等. 2017. 小麦TaZTP29基因克隆及耐盐性鉴定[J]. 农业生物技术学报, 25(10): 1555-1565. (Li S, Li M, Liu Z H, et al.2017. Cloning and salt tolerance identification of TaZTP29 gene in wheat (Triticum aestivum)[J]. Journal of Agricultural Biotechnology, 25(10): 1555-1565.) 5 柳延涛, 陈寅初, 李万云, 等. 2011. 作物抗旱生理生化特性研究进展[J]. 耕作与栽培, (2): 6-7. (Liu Y T, Chen Y C, Li W Y, et al. 2011. Advances in research on physiological and biochemical characteristics of crop drought resistance[J]. Tillage and Cultivation, (2): 6-7.) 6 汪颖. 2011. 我国小麦抗旱性研究进展[J]. 园艺与种苗, (2): 95-97. (Wang Y. 2011. Research progress on drought resistance of wheat in China[J]. Horticulture and Seed, (2): 95-97.) 7 吴迪, 陈林, 张向展, 等. 2016. 小麦TaFKBP62c-2B基因克隆, 组织表达及亚细胞定位[J]. 农业生物技术学报, 24(7): 997-1007. (Wu D, Chen L, Zhang X Z, et al.2016. Cloning, expression and subcellular localization of TaFKBP62c-2B gene in wheat (Triticum aestivum)[J]. Journal of Agricultural Biotechnology, 24(7): 997-1007.) 8 张保才, 周奕华. 2015. 植物细胞壁形成机制的新进展[J]. 中国科学:生命科学, 45(6): 544-556. (Zhang B C, Zhou Y H.2015. New progress in plant cell wall formation mechanism[J]. Science in China, 45(6): 544-556.) 9 邹琦. 2000. 植物生理生化实验指导[M]. 北京: 中国农业出版社, 62-63,134-137. (Zou Q.2000. Plant physiology and biochemistry experiment guide[M]. Beijing: China Agriculture Press, 62-63, 134-137.) 10 Appels R, Eversole K, Feuillet C, et al.2018. Shifting the limits in wheat research and breeding using a fully annotated reference genome[J]. Science, 361(6403): eaar7191. 11 Ariizumi T, Amagai M, Shibata D, et al.2002. Comparative study of promoter activity of three anther-specific genes encoding lipid transfer protein, xyloglucan endotransglucosylase/hydrolase and polygalacturonase in transgenic Arabidopsis thaliana[J]. Plant Cell Reports, 21(1): 90-96. 12 Asa M. Kallas, Piens K, Stuart E. Denman, et al.2005. Enzymatic properties of native and deglycosylated hybrid aspen (Populus tremula×tremuloides) xyloglucan endotransglycosylase 16A expressed in Pichia pastoris[J]. Biochemical Journal, 390(1): 105-113. 13 Becnel J, Natarajan M, Kipp A, et al.2006. Developmental expression patterns of Arabidopsis XTH genes reported by transgenes and gene vestigator[J]. Plant Molecular Biology, 61(3): 451-467. 14 Catala C, Rose J K C, York W S, et al.2001. Characterization of a tomato xyloglucan endotransglucosylase gene that is down- regulated by auxin in etiolated hypocotyls[J]. Plant Physiology, 127: 1180-1192. 15 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]. Febs Letters, 580(13): 3136-3144. 16 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. 17 Cosgrove D J.2005. Growth of the plant cell wall[J]. Nature Reviews Molecular Cell Biology, 6: 850-861. 18 Cui X Y, Du Y T, Fu J, et al.2018. Wheat CBL-interacting protein kinase 23 positively regulates drought stress and ABA responses[J]. BMC Plant Biology, 18(1): 93 19 Eklof J M, Brumer H.2010. The XTH gene family: An update on enzyme structure, function, and phylogeny in xyloglucan remodeling[J]. Plant Physiology, 153(2): 456-466. 20 Geisler-Lee J, Geisler M, Coutinho P M, et al.2006. Poplar carbohydrate-active enzymes gene identification and expression analyses[J]. Plant Physiology, 140(3): 946-962. 21 Genovesi V, Fornale S, Fry S C, et al.2008. ZmXTH1, a new xyloglucan endotransglucosylase/hydrolase in maize, affects cell wall structure and composition in Arabidopsis thaliana[J]. Journal of Experimental Botany, 59(4): 875-889. 22 Han Y, Ban Q, Li H, et al.2016. DkXTH8, a novel xyloglucan endotransglucosylase/hydrolase in persimmon, alters cell wall structure and promotes leaf senescence and fruit postharvest softening[J]. Scientific Reports, 6(1): 39155. 23 Han Y, Wang W, Sun J, et al.2013. Populus euphratica XTH overexpression enhances salinity tolerance by the development of leaf succulence in transgenic tobacco plants[J]. Journal of Experimental Botany, 64(14): 4225-4238. 24 Harada T, Torii Y, Morita S, et al.2011. Cloning, characterization, and expression of xyloglucan endotransglucosylase/hydrolase and expansion genes associated with petal growth and development during carnation flower opening[J]. Journal of Experimental Botany, 62(2): 815-823. 25 Hyodo H, Yamakawa S, Takeda Y, et al.2003. Active gene expression of a xyloglucan endotransglucosylase/hydrolase gene, XTH9, in inflorescence apices is related to cell elongation in Arabidopsis thaliana[J]. Plant Molecular Biology, 52(2): 473-482. 26 Kurasawa K, Matsui A, Yokoyama R, et al.2008. The AtXTH28 gene, a xyloglucan endotransglucosylase/hydrolase, is involved in automatic self-pollination in Arabidopsis thaliana[J]. Plant and Cell Physiology, 50(2): 413-422. 27 Liu Y B, Lu S M, Zhang J F, et al.2007. A xyloglucan endotransglucosylase/hydrolase involves in growth of primary root and alters the deposition of cellulose in Arabidopsis[J]. Planta, 226(6): 1547-1560. 28 Lu W J, Nakano R, Kubo Y, et al.2004. Cloning and expression analysis of an XET cDNA in the peel and pulp of banana fruit ripening and softening[J]. Plant Journal,3(3): 355-362. 29 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]. Plant Journal, 42(4): 525-534. 30 Montserrat Saladié, Rose J K C, 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]. Plant Journal, 47(2): 282-295. 31 Nishitani K.1995. Endo-xyloglucan transferase, a new class of transferase involved in cell wall construction[J]. Plant Research, 108(1089): 137-148. 32 Ookawara R.2005. Expression of α-expansin and xyloglucan endotransglucosylase /hydrolase genes associated with shoot elongation enhanced by anoxia, ethylene and carbon dioxide in arrowhead (Sagittaria pygmaea Miq.) Tubers[J]. Annals of Botany, 96(4): 693-702. 33 Potter I, Fry S C.1994. Changes in xyloglucan endotransglycosylase (XET) activity during hormone-induced growth in lettuce and cucumber hypocotyls and spinach cell suspension cultures[J]. Journal of Experimental Botany, 45: 1703-1710. 34 Rose J K C, Braam J, Fry S C.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. 35 Sasidharan R, Chinnappa C C, Staal M, et al.2010. Light quality-mediated petiole elongation in Arabidopsis during shade avoidance involves cell wall modification by xyloglucan endotransglucosylase/hydrolases[J]. Plant Physiology, 154(11): 978-90. 36 Shin Y K, Yum H, Kim E S, et al.2006. BcXTH1, a Brassica campestris homologue of Arabidopsis XTH9, is associated with cell expansion[J]. Planta, 224(1): 32-41. 37 Soga K, Wakabayashi K, Kamisaka S, et al.2010. Effects of hypergravity on expression of XTH genes in azuki bean epicotyls[J]. Plant Physiology, 131(2): 332-340. 38 van Sandt V S T, Guisez Y, Verbelen J P, et al.2006. Analysis of a xyloglucan endotransglycosylase/hydrolase (XTH) from the lycopodiophyte Selaginella kraussiana suggests that XTH sequence characteristics and function are highly conserved during the evolution of vascular plants[J]. Journal of Experimental Botany, 57(12): 2909-2922. 39 Yang J L, Zhu X F, Peng Y X, et al.2011. Cell wall hemicellulose contributes significantly to aluminum adsorption and root growth in Arabidopsis[J]. Plant Physiology, 155(4): 1885-1892. 40 Yokoyama R, Rose J K C, Nishitani K.2004. A surprising diversity and abundance of xyloglucan endotransglucosylase/hydrolases in rice. Classification and expression analysis[J]. Plant Physiology, 134(3): 1088. 41 Yokoyama R.2001. A comprehensive expression analysis of all members of a gene family encoding cell-wall enzymes allowed us to predict cis-regulatory regions involved in cell-wall construction in specific organs of Arabidopsis[J]. Plant and Cell Physiology, 42(10): 1025-1033. |
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