Cloning and Bioinformatic Analysis of RcPIP2 from Racomitium canescens

doi:10.7668/hbnxb.201751698

Abstract

Abstract: Aquaporins (AQPs) are small transmembrane proteins, and play an important role in tolerance of plants to abiotic stresses. To study the biological functions of aquaporin genes from Racomitrium canescens, a novel plasma membrane intrinsic protein gene of aquaporin family designated RcPIP2 was isolated from the drought transcriptome sequences. The bioinformation results showed that the cDNA full length of RcPIP2 gene was 1 267 bp,which contained an open reading frame of 807 bp and encoded a protein of 268 amino acid residues. It was predicted that the molecular weight of the protein was 28.3 ku, and its theoretical isoelectric point was 6.64, belonging to a stable and hydrophobic protein with no signal peptide. It was a non-secretory protein. RcPIP2 exhibited a typical structure with six transmembrane domains, and had the highly conserved sequence HINPAVTFG of membrane intrinsic protein (MIP) family and two highly conserved Asn-Pro-Ala (NPA) peptides. Its secondary structure contained 37.31% α-helixes, 20.9% β-sheets, and 38.43% random curls. The three-dimensional structure of the RcPIP2 protein was a typical tetramer consisting of four cylindrical subunits and the middle channel. Phylogenetic tree analysis indicated that the protein had the closest genetic relationship with PIP2 protein of Physcomitrella patens, being grouped to the same class. It was inferred that RcPIP2 was one of the aquapion family members.

Key words: Racomitrium canescens, RcPIP2, Aquaporin, Cloning, Bioinformatic analysis

CLC Number:

ZHANG Meijuan, XU Honghong, MA Tianyi, QIAN Pengzhi, SHA Wei. Cloning and Bioinformatic Analysis of RcPIP2 from Racomitium canescens[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 30-36. doi: 10.7668/hbnxb.201751698.

/ / Recommend / Download Citations

URL: http://www.hbnxb.net/EN/10.7668/hbnxb.201751698

http://www.hbnxb.net/EN/Y2019/V34/I5/30

References

[1] Grondin A, Mauleon R, Vadez V, Henry A. Root aquaporins contribute to whole plant water fluxes under drought stress in rice (Oryza sativa L.)[J]. Plant Cell and Environment,2016, 39(2):347-365. doi:10.1111/pce.12616.
[2] Johanson U, Karlsson M, Johansson I, Gustavsson S, Sjövall S, Fraysse L, Weig A R, Kjellbom P. The complex set of genes encoding major intrinsic proteins in Arabidopsis provides a framework for a new nomenclature for major intrinsic proteins in plants[J]. Plant Physiology, 2001, 126(4):1358-1369. doi:10.1104/pp.126.4.1358.
[3] Chaumont F, Barrieu F, Wojcik E, Chrispeels M J, Jung R. Aquaporins constitute a large and highly divergent protein family in maize[J]. Plant Physiology, 2001, 125(3):1206-1215.doi:10.1104/pp.125.3.1206.
[4] Sakurai J, Ishikawa F, Yamaguchi T, Uemura M, Maeshima M. Identification of 33 rice aquaporin genes and analysis of their expression and function[J]. Plant Cell Physiology, 2005, 46(9):1568-1577. doi:10.1093/pcp/pci172.
[5] Fouquet R, Léon C, Ollat N, Barrieu F. Identification of grapevine aquaporins and expression analysis in developing berries[J]. Plant Cell Reports, 2008, 27(9):1541-1550. doi:10.1007/s00299-008-0566-1.
[6] Sade N, Vinocur B J, Diber A, Shatil A, Ronen G, Nissan H, Wallach R, Karchi H, Moshelion M. Improving plant stress tolerance and yield production:is the tonoplast aquaporin SlTIP2; 2 a key to isohydric to anisohydric conversion?[J]. New Phytologist Trust, 2009, 181(3):651-661. doi:10.1111/j.1469-8137.2008.02689.x.
[7] Park W, Scheffler B E, Bauer P J, Campbell B T. Identification of the family of aquaporin genes and their expression in upland cotton (Gossypium hirsutum L.)[J]. BMC Plant Biology, 2010, 10(1):142.doi:10.1186/1471-2229-10-142.
[8] Zhang D Y, Ali Z, Wang C B, Xu L, Yi J X, Xu Z L, Liu X Q, He X L, Huang Y H, Khan I A, Trethowan R M, Ma H X. Genome-wide sequence characterization and expression analysis of major intrinsic proteins in soybean (Glycine max L.)[J]. PLoS One, 2013, 8(2):e56312.doi:10.1371/journal.pone.0056312.
[9] 吴向阳, 程朝泽, 吕高强, 王心宇. 芝麻AQP家族的全基因组序列鉴定及其特征分析[J]. 中国农业科学, 2016, 49(10):1844-1858. doi:10.3864/j.issn.0578-1752.2016.10.002.
W X Y, Cheng C Z, Lǘ G Q, Wang X Y. Identification and characterization of the AQP gene family in sesame[J]. Scientia Agricultura Sinica, 2016, 49(10):1844-1858.
[10] Zardoya R. Phylogeny and evolution of the major intrinsic protein family[J]. Biology of the Cell, 2005, 97(6):397-414. doi:10.1042/BC20040134.
[11] Gustavsson S, Lebrun A S, Nordén K, Chaumont F, Johanson U. A novel plant major intrinsic protein in Physcomitrella patens most similar to bacterial glycerol channels[J]. Plant Physiology, 2005, 139(1):287-295. doi:10.1104/pp.105.063198.
[12] Danielson J Å, Johanson U. Unexpected complexity of the aquaporin gene family in the moss Physcomitrella patens[J]. BMC Plant Biology, 2008, 8(1):45. doi:10.1186/1471-2229-8-45.
[13] 王星, 张纪龙, 冯秀秀, 李洪杰, 张根发. 植物质膜水通道蛋白转运及逆境胁迫响应的分子调控机制[J]. 遗传, 2017, 39(4):293-301. doi:10.16288/j.yczz.16-290.
Wng X, Zhang J L, Feng X X, Li H J, Zhang G F. The molecular mechanisms of plant plasma membrane intrinsic proteins trafficking and stress response[J].Hereditas,2017, 39(4):293-301.
[14] 江林娟, 陈春华, 颜旭, 杨世民. 植物水通道蛋白的干旱应答机制研究进展[J]. 广西植物, 2018, 38(5):672-680. doi:10. 11931/guihaia. gxzw201705032.
Jang L J, Chen C H, Yan X, Yang S M. Research progress on responsive mechanism of aquaporins to drought stress in plants[J]. Guihaia, 2018, 38(5):672-680.
[15] 包珠拉太, 高丽, 王锁民. 植物水通道蛋白及其生理功能[J]. 植物生理学报, 2017, 53(7):1117-1178. doi:10.13592/j.cnki.ppj.2017.0076.
Bo Z L T, Gao L, Wang S M. Physiological functions of plant aquaporin[J]. Plant Physiology Journal, 2017, 53(7):1171-1178.
[16] 何勇清, 方佳, 余敏芬, 方仲相, 江波, 潘寅辉, 郑炳松. 植物质膜内在水通道蛋白PIPs的分子生物学研究进展[J]. 浙江农林大学学报, 2012, 29(3):446-452.doi:10.3969/j.issn.2095-0756.2012.03.020.
H Y Q, Fang J, Yu M F, Fang Z X, Jiang B, Pan Y H, Zheng B S. Advances in molecular biology for plasma membrane intrinsic proteins (PIPs):a review[J]. Journal of Zhejiang Agriculture and Forestry University, 2012, 29(3):446-452.
[17] 李菲, 颜培玲, 张文娥, 潘学军. 野生毛葡萄水通道蛋白基因VhPIPs 的克隆与组织特异性表达研究[J]. 园艺学报, 2016, 43(12):2304-2314. doi:10.16420/j.issn.0513-353x.2016-0265.
L F, Yan P L, Zhang W E, Pan X J. Cloning and tissue specificity expression of the aquaporin genes from wild Vitis quinquangularis[J].Acta Horticulturae Sinica, 2016, 43(12):2304-2314.
[18] Ding L, Gao C M, Li Y R, Li Y, Zhu Y Y, Xu G H, Shen Q R, Kaldenhoff R, Kai L, Guo S W. The enhanced drought tolerance of rice plants under ammonium is related to aquaporin (AQP)[J]. Plant Science, 2015, 234:14-21. doi:10.1016/j.plantsci.2015.01.016.
[19] 周菲, 程晨晨, 王然, 杨英杰, 宋健坤. 杜梨干旱胁迫相关水通道蛋白基因的筛选与克隆[J]. 北方园艺, 2019,43(2):48-54. doi:10.11937/bfyy.20182123.
Zou F, Cheng C C, Wang R, Yang Y J, Song J K. Screening and cloing of drought stress related aquaporin genes in Pyrus betulifolia Bunge[J]. Northern Horticultur, 2019,43(2):48-54.
[20] Xu Y, Hu W, Liu J H, Zhang J B, Jia C H, Miao H X, Xu B Y, Jin Z Q. A banana aquaporin gene, MaPIP1; 1, is involved in tolerance to drought and salt stresses[J]. BMC Plant Biology, 2014, 14(1):59. doi:10.1186/1471-2229-14-59.
[21] 蔡琼, 丁贵杰, 文晓鹏. 马尾松水通道蛋白PmPIP1 基因克隆及在干旱胁迫下的表达分析[J]. 浙江农林大学学报, 2016, 33(2):191-200. doi:10.11833/j.issn.2095-0756.2016.02.002.
Ci Q, Ding G J, Wen X P. Cloning of the PmPIP1 gene from Pinus massoniana and its expression with drought stress[J]. Journal of Zhejiang Agriculture and Forestry University, 2016, 33(2):191-200.
[22] Zhuang L L,Liu M X,Yuan X Y,Yang Z M, Huang B R. Physiological effects of aquaporin in regulating drought tolerance through overexpressing of Festuca arundinacea aquaporin gene FaPIP2; 1[J]. Journal of the American Society for Horticultural Science, 2015, 140(5):404-412.doi:10.21273/JASHS.140.5.404.
[23] Li J T, Yu G, Sun X H, Liu Y Z, Liu J L, Zhang X H, Jia C G, Pan H Y. AcPIP2, a plasma membrane intrinsic protein from halophyte Atriplex canescens, enhances plant growth rate and abiotic stress tolerance when overexpressed in Arabidopsis thaliana[J]. Plant Cell Reports, 2015, 34(8):1401-1415. doi:10.1007/s00299-015-1796-7.
[24] 沙伟, 张梅娟, 刘博, 徐红红. 东亚砂藓bZIP转录因子RjbZIP 基因的克隆及表达分析[J]. 华北农学报, 2015, 30(4):13-20. doi:10.7668/hbnxb.2015.04.003.
Sa W, Zhang M J, Liu B, Xu H H. Cloning and expression analysis of a bZIP transcription factor gene, RjbZIP, from Racomitrium japonicum[J]. Acta Agriculturae Boreali-Sinica, 2015, 30(4):13-20.
[25] 张萍, 白学良, 钟秀丽. 苔藓植物耐旱机制研究进展[J]. 植物学通报, 2005, 22(1):107-114.doi:10.3969/j.issn.1674-3466.2005.01.017.
Zang P, Bai X L, Zhong X L. Advances in the desiccation tolerance of mosses[J]. Chinese Bulletin Botany, 2005, 22(1):107-114.
[26] 王彦梅, 张正斌, 柴团耀, 徐萍, 赵鸿彬.植物水通道蛋白研究进展[J].西北农林科技大学学报(自然科学报), 2005, 33(11):106-112. doi:10.3321/j.issn:1671-9387.2005.11.021.
Wng Y M, Zhang Z B, Chai T Y, Xu P, Zhao H B. Advances in the studies on plant aquaporins[J]. Journal of Northwest Science-Technology University of Agriculture and Foresty(Natural Science Edition), 2005, 33(11):106-112.

Please choose a citation manager

Content to export