雪里蕻非特异脂质转运蛋白基因的克隆与分析

doi:10.7677/ynzwyj201312150

摘要/Abstract

摘要：

非特异脂质转运蛋白是植物生命活动中一类重要的活性蛋白，这类蛋白在植物的抗性和防御中行使着重要的功能。近年来研究发现这类蛋白还与植物的有性生殖密切相关。通过已得到的普通白菜的脂质转运蛋白基因BcMF15的核苷酸序列，在其基因全长两侧设计引物，从雪里蕻中克隆得到该类活性蛋白基因，命名为BjLTP (登陆号: EU082009)。该基因全长650bp，不含内含子。不同组织的表达特征分析发现，BjLTP在雪里蕻的花蕾、开放的花中特异表达，推测BjLTP可能与花粉的发育有关。蛋白质特征预测及蛋白序列结构分析发现BjLTP是一个跨膜蛋白，具有显著的疏水区。序列同源比对表明该基因与白花芥蓝、拟南芥等的脂质转运蛋白基因有很高的相似性，证明BjLTP是LTP家族的成员之一。

关键词: 雪里蕻, 脂质转运蛋白, 基因

Abstract:

Nonspecific Lipid transfer proteins (nsLTPs) were a group of important active proteins in in higher plants. Their function has been suggested to have a role in plant resistance and defense response. According to recent studies, LTPs play an important role in plant reproduction and development. In this study, BjLTP was cloned from Brassica juncea var. multiceps based on BcMF15 from Brassica campestris ssp. chinensis var. communis BjLTP was 650bp (EU082009) in length with no intron. RTPCR analysis showed that BjLTP was expressed in flower buds and open flower but not other tissues. The gene may act as nsLTPs involving in pollen development. Protein characteristics and sequence structure were predicted, that it was a transmembrane protein with obvious hydrophobic region. BjLTP was identified as a Lipid transfer protein gene according to its high identity with Brassica oleracea (MF15bh) and Arabidopsis thaliana LTP.

Key words: Brassica juncea var. multiceps, Lipid transfer proteins, Gene

中图分类号:

Q 943.2

田爱梅. 雪里蕻非特异脂质转运蛋白基因的克隆与分析[J]. Plant Diversity, 2013, 35(5): 563-568.

TIAN Ai-Mei. Isolation and Molecular Characterization of nsLTPLike Gene from Brassica juncea var. multiceps (Cruciferae)[J]. Plant Diversity, 2013, 35(5): 563-568.

参考文献

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: 90—96
Chae K, Kiehlich CA, Morikis DA et al., 2009. Gainoffunction mutation of Arabidopsis lipid transfer protein 5 disturbs pollen tube tip growth and fertilization［J］. The Plant Cell, 21: 3902—3914
Chen L (陈玲), Zhang H (张颢), Qiu XQ (邱显钦) et al., 2012. Cloning and analysis of NBSLRR type disease resistance gene analogs from Rosa rubusin in Yunnan［J］. Plant Diversity and Resources (植物分类与资源学报), 34 (1): 56—62
Crossley SJ, Greenland AJ, Dickinson HG, 1995. The characterization of tapetumspecific cDNAs isolated from a Lilium henryi L. meiocyte subtractive cDNA library［J］. Planta, 196: 523—529
EiShesawi AM, Elseehy MM, Hedgcoth C, 2011. Isolation and sequence analysis of wheat tissuespecific cDNAs by differential display［J］. Plant Molecular Biology Reporter, 29 (1): 135—148
Gao G (郜刚), Ren CH (任彩虹), Jin LP (金黎平) et al., 2008. Cloning, expression and characterization of a nonspecific lipid transfer protein gene from potato［J］. Acta Agronomica Sinica (作物学报), 34 (9): 1510—1517
Gao NB (高乃波), Liu XG (刘晓光), Shi ML (施曼玲), 2009. Establishment of efficient regeneration system of Brassica juncea var. multiceps［J］. Acta Agriculturae Zhejiangensis (浙江农业学报), 21 (2): 149—153
Hang MD, Wei FJ, Wu CC et al., 2009. Analyses of advanced rice anther transcriptomes reveal global tapetum secretory functions and potential proteins for lipid exine formation［J］. Plant Physiology, 149: 694—707
Jiang Q (蒋倩), Wang F (王枫), Hou XL (侯喜林) et al., 2012. Cloning and expression pattern analysis of nsLTP gene in Celery［J］. Acta Horticulturae Sinica (园艺学报), 39 (7): 1293—1302
Kato H, Xie G, Sato Y et al., 2010. Isolation of antherspecific gene promoters suitable for transgene expression in rice［J］. Plant Molecular Biology Reporter, 28: 381—387
Liu M (刘梅), Sheng H (生华), Hua WP (化文平) et al., 2011. Cloning and expression analysis of a nonspecific lipid transfer protein gene (SmLTP1) from Salvia miltiorrhiza Bunge［J］. Plant Physiology Journal (植物生理学报), 47 (1): 63—68
Liu Q (刘群), Ma J (马婧), Zhu QL (祝钦泷) et al., 2009. Expression analysis of three nonspecific lipid transfer protein (nsLTP) genes in response to abiotic stresses in Chimonanthus praecox［J］. Journal of Agricultural Biotechnology (农业生物技术学报), 17 (6): 1027—1034
Liu ZY (刘志勇), Ye XL (叶雪凌), Li CY (李承彧)et al., 2011. Cloning and characterization of a genic male sterilityrelated gene BrLTP1 in Chinese Cabbage［J］. Acta Horticulturae Sinica (园艺学报), 38 (2): 343—352
Park SY, Jauh GY, Mollrt JC et al., 2000. A lipid transferlike protein is necessary for lily pollen tube adhesion to an in vitro stylar matrix［J］. The Plant Cell, 12: 51—163
Tian AM (田爱梅),Cao JS (曹家树), 2008. Lipid transfer proteins in plants［J］. Chinese Journal of Cell Biology (细胞生物学杂志), 30: 483—488
Tian AM, Cao JS, Huang L et al., 2009. Characterization of a male sterile related gene BcMF15 from Brassica campestris ssp. Chinensis［J］. Molecular Biology Reports, 36: 307—314
Wang LP (王玲平), Cao JS (曹家树), Ye WZ (叶纨芝) et al., 2005. Cloning and evolutionary analysis of homologous sequences of CYP86MF gene in Cruciferae［J］. Hereditas (遗传), 27 (3): 395—402
Yu HF (虞慧芳), Zhong XM (钟新民), Gu HH (顾宏辉) et al., 2007. Obtaining microsporecultured plants in potherb mustard, Brassica juncea var. multiceps, by spacebreeding satellite［J］. Acta Agriculturae Zhejiangensis (浙江农业学报), 19 (5): 356—359
Zhang DS, Liang WQ, Yin CS et al., 2010. OsC6, encoding a lipid transfer protein, is required for postmeiotic anther development in rice［J］. Plant Physiology, 154: 149—162

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