橡胶树胶乳橡胶粒子死皮相关蛋白的鉴定及分析

袁坤; 周雪梅; 王真辉; 杨礼富

doi:10.3969/j.issn.1000-2006.2014.01.007

袁坤,周雪梅,王真辉,杨礼富

南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (01) : 36-40.

PDF(1779722 KB)

国家林草科技领军期刊
中国精品科技期刊
中国高校百佳科技期刊
江苏省新闻出版政府奖期刊奖
RCCSE林学权威期刊（A+）
CSCD核心期刊
Scopus数据库收录期刊
中文核心期刊
SCD核心期刊

作者加群：102861116

微信公众号：南京林业大学学报

高级检索

PDF(1779722 KB)

南京林业大学学报（自然科学版） ›› 2014, Vol. 38 ›› Issue (01) : 36-40. DOI: 10.3969/j.issn.1000-2006.2014.01.007

研究论文

橡胶树胶乳橡胶粒子死皮相关蛋白的鉴定及分析

袁坤¹,周雪梅^1,2,王真辉¹,杨礼富^1*

作者信息 +

Identification and analysis of latex rubber particle proteins related with tapping panel dryness in Hevea brasiliensis

YUAN Kun¹, ZHOU Xuemei^1,2, WANG Zhenhui¹, YANG Lifu^1*

Author information +

文章历史 +

摘要

橡胶树死皮是影响天然橡胶产量的重要因子之一。采用双向凝胶电泳对健康树和死皮树橡胶粒子中差异表达的蛋白进行分离,分析获得35个差异表达的蛋白点,通过基质辅助激光解析电离飞行时间质谱(MALDI-TOF MS)分析及搜索Uniprot rubber数据库后,有13个蛋白点被成功鉴定,其中橡胶延长因子(rubber elongation factor,REF)、法尼基焦磷酸合成酶(Ffarnesyl-diphosphate synthase,FPS)、谷胱甘肽过氧化酶(glutathione peroxidase,GPX)、谷胱甘肽还原酶(glutathione reductase,GR)、翻译控制肿瘤蛋白(translationally controlled tumor protein,TCTP)、热激蛋白(heat shock protein, HSP)等主要蛋白参与了橡胶的生物合成、活性氧代谢及细胞凋亡过程。说明橡胶的生物合成、活性氧代谢及细胞凋亡途径可能是橡胶树死皮发生的关键调控途径。

Abstract

The tapping panel dryness(TPD)in Hevea brasiliensis is one of the important factors influencing natural rubber production. In order to distinguish and identify the differentially expressed proteins of latex rubber particles in healthy and TPD trees, the two-dimensional electrophoresis(2-DE)was used. 35 differentially expressed protein spots were obtained by software analysis. Subjecting to MALDI-TOF MS following by searching for Uniprot rubber database, 13 protein spots were successfully identified, Of them, REF, FPS, GPX, GR, TCTP and HSP were mainly involed in the rubber biosynthesis, reactive oxygen metabolism and cell apoptosis processes. This study further showed that the pathways of the rubber biosynthesis, reactive oxygen metabolism and cell apoptosis were possibly the key regulated ones of TPD and which would provide a theoretical basis for elucidating the TPD mechanism in Hevea brasiliensis.

引用本文

EndNote

Ris (Procite)

Bibtex

导出引用

袁坤,周雪梅,王真辉,杨礼富. 橡胶树胶乳橡胶粒子死皮相关蛋白的鉴定及分析[J]. 南京林业大学学报（自然科学版）. 2014, 38(01): 36-40 https://doi.org/10.3969/j.issn.1000-2006.2014.01.007

YUAN Kun, ZHOU Xuemei, WANG Zhenhui, YANG Lifu. Identification and analysis of latex rubber particle proteins related with tapping panel dryness in Hevea brasiliensis[J]. JOURNAL OF NANJING FORESTRY UNIVERSITY. 2014, 38(01): 36-40 https://doi.org/10.3969/j.issn.1000-2006.2014.01.007

中图分类号： S794.1

参考文献

[1] Cornish K. Similarities and differences in rubber biochemistry among plant species [J]. Phytochemistry, 2001, 57(7):1123-1134.
[2] Cornish K, Wood D F, Windle J J. Rubber particles from four different species, examined by transmission electron microscopy and electron-paramagnetic-resonance spin labeling, are found to consist of a homogeneous rubber core enclosed by a contiguous, monolayer biomembrane [J]. Planta, 1999, 210(1): 85-96.
[3] Wood D F, Cornish K. Microstructure of purified rubber particles [J]. Int J Plant Sci, 2000, 161(3):435-445.
[4] Dennis M S, Light D R. Rubber elongation factor from Hevea brasiliensis,identification,characterization,and role in rubber biosynthesis [J]. J Biol Chem, 1989, 264(31):18608-18617.
[5] 段翠芳, 聂智毅, 曾日中. 橡胶粒子膜蛋白双向电泳体系的建立和质谱初步分析 [J]. 热带作物学报, 2006, 27(3):22-29. Duan C F, Nie Z Y, Zeng R Z. Establishment of 2-DE system and primary analyses on the membrane proteins of rubber particles in Hevea brasiliensis by MALDI-TOF [J]. Chinese Journal of Tropical Crops, 2006, 27(3):22-29.
[6] 陈春柳, 闫洁, 邓治, 等. 橡胶树死皮橡胶粒子膜蛋白差异分析与初步鉴定 [J]. 中国农学通报, 2010, 26(5):304-308. Chen C L, Yan J, Deng Z, et al. Differential analysis of 2-DE profiles and preliminary identification of lutoids proteins from TPD tree in Hevea brisiliensis [J]. Chinese Agricultural Science Bulletin, 2010, 26(5):304-308.
[7] 周雪梅, 杨礼富, 王真辉, 等. 橡胶树胶乳黄色体死皮相关蛋白的鉴定及分析 [J]. 西南农业学报, 2012, 25(6):2093-2097. Zhou X M, Yang L F, Wang Z H, et al. Identification and analysis of proteins in latex lutoid associated with tapping panel dryness of Hevea brasiliensis [J]. Southwest China Journal of Agricultural Sciences, 2012, 25(6):2093-2097.
[8] Blum H, Beier H, Hans J. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels [J]. Electrophoresis, 1987, 17(8):93-99.
[9] Fernandez J, Gharahdaqhi F, Mische S M. Routine identification of proteins from sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)gel or polyvinyl difluoride membranes using matrix assisted laser desorption/ionization-time of flight-mass spectrometry(MALDI-TOF-MS)[J]. Electrophoresis, 1998, 19(6):1036-1045.
[10] Gharahdaghi F, Weinberg C R, Meagher D A, et al. Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity [J]. Electrophoresis, 1999, 20(3): 601-605.
[11] 周雪梅, 杨礼富, 王真辉, 等. 橡胶树胶乳C-乳清死皮相关蛋白的鉴定及分析 [J]. 南京林业大学学报:自然科学版, 2012, 36(5):37-41. Zhou X M, Yang L F, Wang Z H, et al. Identification and analysis of proteins in the latex C-serum related to the tapping panel dryness in Hevea brasiliensis [J]. Journal of Nanjing Forestry University:Natural Sciences Edition, 2012, 36(5):37-41.
[12] 张福城, 陈守才. 巴西橡胶树天然橡胶生物合成中关键酶及相关基因研究进展 [J]. 热带农业科学, 2006, 26(1):42-46. Zhang F C, Chen S C. Progress on the key enzymes and related genes of natural rubber biosynthesis pathway from Hevea brasiliensis [J]. Chinese Journal of Tropical Agriculture, 2006, 26(1):42-46.
[13] 段翠芳, 曾日中, 黎瑜. 激素对巴西橡胶树橡胶生物合成的调控 [J]. 热带农业科学, 2004, 10(5):61-67. Duan C F, Zeng R Z, Li Y. Regulation of plant hormones on biosynthesis of natural rubber in Hevea brasiliensis [J]. Chinese Journal of Tropical Agriculture, 2004, 10(5):61-67.
[14] Li D, Deng Z, Chen C, et al. Identification and characterization of genes associated with tapping panel dryness from Hevea brasiliensis latex using suppression subtractive hybridization[J]. BMC Plant Biology, 2010, 10(1): 140-151.
[15] Noctor G, Gomez L, Vanacker H, et al. Interactions between biosynthesis,compartmentation and transport in the control of glutathione homeostasis and signaling [J]. J Exp Bot, 2002, 53: 1283-1304.
[16] Potters G, Horemans N, Bellone S, et al. Dehydroascorbate influences the plant cell cycle through a glutathione-independent reduction mechanism [J]. Plant Physiol, 2004, 134(4): 1479-1487.
[17] Chew O, Whelan J, Millar A H. Molecular definition of the ascorbate-glutathione cycle in Arabidopsis mitochondria reveals dual targeting of antioxidant defenses in plants [J]. J Biol Chem, 2003, 278(47): 46869-46877.
[18] 宫维嘉, 金赞敏, 王长海. 海水胁迫下库拉索芦荟南盐1号抗氧化酶活力的变化 [J]. 江苏农业科学, 2006(6):348-350. Gong W J, Jin Z M, Wang C H. Changes of antioxidant enzyme activity in aloe variety Nanyan No. 1(Aloe vera)under seawater stress [J]. Jiangsu Agricultural Sciences, 2006(6):348-350.
[19] 陈军文, 曹坤芳. 三叶橡胶光合作用能力和抗氧化系统以及单萜类物质对茉莉酸的响应 [J]. 植物研究, 2008,28(1):48-53. Chen J W, Cao K F. Reponses of photosynthetic capacities,antioxidant activities and monoterpene biosynthesis in leaves of Hevea brasiliensis tree to jasmonic acid [J]. Bulletin of Botanical Research, 2008, 28(1):48-53.
[20] 闫洁, 陈守才, 夏志辉. 橡胶树死皮病胶乳C-乳清差异表达蛋白质的筛选与鉴定 [J]. 中国生物工程杂志, 2008, 28(6):28-36. Yan J, Chen S C, Xia Z H. Screening and identification of differential expressed proteins in c-serum on latex of tapping panel dryness(TPD)in rubber tree(Hevea brasiliensis Muell. Arg.)[J]. China Biotechnology, 2008, 28(6):28-36.
[21] Venkatachalam P, Thulaseedharan A, Raghothama K. Identification of expression profiles of tapping panel dryness(TPD)associated genes from the latex of rubber tree(Hevea brasiliensis Muell. Arg.)[J]. Planta, 2007, 226(2):499-515.
[22] Rho S B, Lee J H, Park M S, et al. Anti-apoptotic protein TCTP controls the stability of the tumor suppressor p53 [J]. FEBS Lett, 2011, 585(1): 29-35.
[23] Hua G, Zhang Q, Fan Z. Heat shock protein 75(TRAP1)antagonizes reactive oxygen species generation and protects cells from granzyme M-mediated apoptosis [J]. J Biol Chem, 2007, 282(28): 20553-20560.
[24] 袁坤, 徐智娟, 王真辉, 等. 橡胶树胶乳死皮相关蛋白的鉴定及分析 [J]. 西北林学院学报, 2012, 27(6): 105-109. Yuan K, Xu Z J, Wang Z H, et al. Identification and analysis of latex proteins related to tapping panel dryness in Hevea brasiliensis [J]. Journal of Northwest Forestry University, 2012, 27(6):105-109.
[25] 赵静雅, 左开井, 秦捷, 等. 油菜烯醇酶基因的克隆与分析 [J]. 植物学报, 2004, 46(10): 1226-1233. Zhao J Y, Zuo K J, Qin J, et al. Molecular cloning and characterization of enolase from oilseed rape(Brassica napus)[J].Acta Botanica Sinica, 2004, 46(10):1226-1233.

基金

收稿日期:2013-03-13 修回日期:2013-07-10
基金项目:中央级公益性科研院所基本业务费专项项目(YWFZX2008-12,2009-12,2010-9,1630022013003); 海南省重点科技项目(90107); 现代农业产业技术体系建设专项资金项目(CARS-34-02A)
第一作者:袁坤,副研究员。*通信作者:杨礼富,研究员。E-mail:ylfri@126.com
引文格式:袁坤,周雪梅,王真辉,等. 橡胶树胶乳橡胶粒子死皮相关蛋白的鉴定及分析[J]. 南京林业大学学报:自然科学版,2014,38(1):36-40.