Isolation and Analysis of Differential Expressed ESTs from Stem Trichomes of Lycopersicon esculentum (Solanaceae)

doi:10.3724/SP.J.1143.2011.11062

Abstract

Abstract:

Glandular trichomes are special organs involved in plant defense response and synthesis of volatile secondary metabolites, analyzing trichome specific expressed sequence tags will help us further understand the specific function of plant trichomes. In this paper, suppression subtractive hybridization（SSH） based on magnetic beads technology was used to isolate differential expressed genes of the glandular trichomes in Lycopersicon esculentum. The differential expressing cDNA library was constructed using the glandular trichomes cDNA as tester and the cDNA from the stem without glandular trichomes as driver. After randomly sequencing 108 differential ESTs, Blast2go program was used to do blastx, functional annotation and metabolism analysis. The results show that most ESTs are related to substance metabolism, response to stress, biotic or abiotic stimulus, and have binding and catalytic function. These differential genes lay the foundation for further research on defense mechanism of the tomato trichomes.

Key words: EST, Glandular trichome, Lycopersicon esculentum, Suppression Subtractive Hybridization（SSH）

摘要：

腺毛是参与植物防御性反应与次生代谢挥发物合成的特异化器官，了解腺毛专一性表达序列标签将有助于我们进一步认识植物腺毛的特异性功能。本文应用磁珠介导的抑制消减杂交方法（SSH），以番茄茎腺毛为检测子（tester），去除腺毛的番茄茎为驱动子（driver），构建番茄茎腺毛差异表达cDNA文库，分离腺毛差异表达基因。随机挑选了108个差异ESTs进行测序，测序结果使用Blast2go程序进行blastx比对、功能注释和KEGG代谢路径分析。结果表明，绝大部分ESTs功能与胁迫响应、物质代谢、生物及非生物刺激反应相关，具有结合、催化功能。这些分离的差异ESTs为进一步研究番茄腺毛的植物防御性机制奠定了基础。

关键词: 表达序列标签, 腺毛, 番茄, 抑制消减杂交

CLC Number:

Q 75

LI Wan-Sha, LIU De-Tuan, YANG Yong-Ping, HU Xiang-Yang. Isolation and Analysis of Differential Expressed ESTs from Stem Trichomes of Lycopersicon esculentum (Solanaceae)[J]. Plant Diversity, 2011, 33(6): 660-666.

李婉莎, 刘德团, 杨永平, 胡向阳. 番茄茎腺毛差异表达序列分离与分析[J]. Plant Diversity, 2011, 33(6): 660-666.

References

Antonious GF, Snyder JC, 2006. Natural products: repellency and toxicity of wild tomato leaf extracts to the twospotted spider mite, Tetranychus urticae Koch［J］. Journal of environmental science and health, Part B: Pesticides, Food Contaminants, and Agricultural Wastes, 41: 43—55
Aziz N, Paiva NL, May GD et al., 2005. Transcriptome analysis of alfalfa glandular trichomes［J］. Planta, 221: 28—38
Boughton AJ, Hoover K, Felton GW, 2005. Methyl jasmonate application induces increased densities of glandular trichomes on tomato, Lycopersicon esculentum［J］. Journal of Chemical Ecology, 31: 2211—2216
BuchananWollaston V, Earl S, Harrison E et al., 2003. The molecular analysis of leaf senescence? a genomics approach［J］. Plant Biotechnology Journal, 1: 3—22
Christensen AB, Cho BHO, Naesby M et al., 2002. The molecular characterization of two barley proteins establishes the novel PR17 family of pathogenesisrelated proteins［J］. Molecular Plant Pathology, 3: 135—144
Conesa A, Gtz S, GarcíaGómez JM et al., 2005. Blast2GO: a universal tool for annotation, visualization and analysis in functional genomics research［J］. Bioinformatics, 21: 3674—3876
Dai X, Wang G, Yang DS et al., 2010. TrichOME: A comparative omics database for plant trichomes［J］. Plant Physiology, 152: 44—54
Freitas JA, Maluf WR, Cardoso gracas das M et al., 2002. Inheritance of foliar zingiberene contents and their relationship to trichome densities and whitefly resistance in tomatoes［J］. Euphytica, 127: 275—287
Higginson T, Li SF, Parish RW, 2003. AtMYB103 regulates tapetum and trichome development in Arabidopsis thaliana［J］. The Plant Journal, 35: 177—192
Kang JH, Shi F, Jones AD et al., 2010. Distortion of trichome morphology by the hairless mutation of tomato affects leaf surface chemistry［J］. Journal of Experimental Botany, 61: 1053—1064
Lnneborg A, Sharma P, Stougaard P, 1995. Construction of subtractive cDNA library using magnetic beads and PCR［J］. Genome Research, 4: 168—176
Li L, Zhao Y, Mccaig BC et al., 2004. The tomato homolog of CORONATINEINSENSITIVE1 is required for the maternal control of seed maturation, jasmonatesignaled defense responses, and glandular trichome development［J］. The Plant Cell Online, 16: 126—143
Luckwill L, 1943. The Genus Lycopersicon: An Historical, Biological, and Taxonomic Survey of the Wild and Cultivated tomatoes［M］. Aberdeen: Aberdeen University Press, Scotland
Martin C, Glover BJ, 2007. Functional aspects of cell patterning in aerial epidermis［J］. Current Opinion in Plant Biology, 10: 70—82
Peiffer M, Tooker JF, Luthe DS et al., 2009. Plants on early alert: glandular trichomes as sensors for insect herbivores［J］. New Phytologist, 184: 644—656
Ranger CM, Backus EA, Winter REK et al., 2004. Glandular trichome extracts from Medicago sativa deter settling by the potato leafhopper Empoasca fabae［J］. Journal of Chemical Ecology, 30: 927—943
Roxas VP, Smith RK, Allen ER et al., 1997. Overexpression of glutathione Stransferase/glutathione peroxidase enhances the growth of transgenic tobacco seedlings during stress［J］. Nature Biotechnology, 15: 988—991
Samanani N, Liscombe DK, Facchini PJ, 2004. Molecular cloning and characterization of norcoclaurine synthase, an enzyme catalyzing the first committed step in benzylisoquinoline alkaloid biosynthesis ［J］. The Plant Journal, 40: 302—313
Schilmiller AL, Schauvinhold I, Larson M et al., 2009. Monoterpenes in the glandular trichomes of tomato are synthesized from a neryl diphosphate precursor rather than geranyl diphosphate［J］. Proceedings of the National Academy of Sciences, 106: 10865—10870
Shangguan XX, Xu B, Yu ZX et al., 2008. Promoter of a cotton fibre MYB gene functional in trichomes of Arabidopsis and glandular trichomes of tobacco［J］. Journal of Experimental Botany, 59: 3533—3542
Shepherd RW, Wagner GJ, 2007. Phylloplane proteins: emerging defenses at the aerial frontline? ［J］. Trends in Plant Science, 12: 51—56
Wang E, Gan S, Wagner GJ, 2002. Isolation and characterization of the CYP71D16 trichomespecific promoter from Nicotiana tabacum L［J］. Journal of Experimental Botany, 53: 1891—1897
Wang G, Tian L, Aziz N et al., 2008. Terpene biosynthesis in glandular trichomes of hop［J］. Plant Physiology, 148: 1254—1266
Xie Z, Kapteyn J, Gang DR, 2008. A systems biology investigation of the MEP/terpenoid and shikimate/phenylpropanoid pathways points to multiple levels of metabolic control in sweet basil glandular trichomes［J］. The Plant Journal, 54: 349—361
Yu T, Li YS, Chen XF et al., 2003. Transgenic tobacco plants overexpressing cotton glutathione Stransferase (GST) show enhanced resistance to methyl viologen［J］. Journal of Plant Physiology, 160: 1305—1311
Zanders ED, Goulden MG, Kennedy TC et al., 2000. Analysis of immune system gene expression in small rheumatoid arthritis biopsies using a combination of subtractive hybridization and highdensity cDNA arrays［J］. Journal of Immunological Methods, 233: 131—140

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