Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata:: V.: Microarray analysis and further characterization of large-scale changes in herbivore-induced mRNAs
Molecular interactions between the specialist herbivore Manduca sexta (Lepidoptera, Sphingidae) and its natural host Nicotiana attenuata:: V.: Microarray analysis and further characterization of large-scale changes in herbivore-induced mRNAs
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DOI：
10.1104/pp.102.018176
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发表时间：
2003-04-01
期刊：
影响因子：
7.4
通讯作者：
Baldwin, IT
中科院分区：
文献类型：
--
作者：
Hui, DQ;Iqbal, J;Baldwin, IT
我们扩展了我们的分析，发生的转录重组时，本地烟草，是由天蛾幼虫的攻击，通过克隆115个转录本mRNA差异显示逆转录聚合酶链反应和消减杂交使用磁珠（SHMB）从M。sexta应答转录组。这些成绩单与其他8个，以前证实是差异调节的北方分析载玻片微阵列上的cDNA斑点，并与Cy 3和Cy 5标记的探针来自植物后2，6，12，和24小时的连续攻击。微阵列分析被证明是一个强大的手段，验证差异表达; 73个克隆基因（63%）差异调节（在相同比例的差异显示逆转录-聚合酶链反应和SHMB程序），其中，24个（32%）与已知基因或推定的蛋白质（更多来自SHMB）相似。分析提供了深入的信号和转录基础的直接和间接防御用于对付食草动物，同时激活水杨酸，乙烯，细胞分裂素，WRKY，MYB，和oxylipin信号通路和牵连萜类，病原体，和细胞壁相关的转录防御反应。这些防御反应需要的资源，可以通过减少四个光合作用相关的转录，增加与蛋白质和核苷酸周转相关的转录，并增加与碳水化合物代谢相关的转录。这种假定的上调的防御相关和下调的生长相关的成绩单发生的背景下，改变转录的RNA结合蛋白，假定的ATP/ADP转运蛋白，伴侣蛋白，组蛋白和水通道蛋白，响应一致的一个主要的代谢重构，强调响应草食动物攻击的复杂性。
We extend our analysis of the transcriptional reorganization that occurs when the native tobacco, Nicotiana attenuata, is attacked by Manduca sexta larvae by cloning 115 transcripts by mRNA differential display reverse transcription-polymerase chain reaction and subtractive hybridization using magnetic beads (SHMB) from the M. sexta-responsive transcriptome. These transcripts were spotted as cDNA with eight others, previously confirmed to be differentially regulated by northern analysis on glass slide microarrays, and hybridized with Cy3- and Cy5-labeled probes derived from plants after 2, 6, 12, and 24 h of continuous attack. Microarray analysis proved to be a powerful means of verifying differential expression; 73 of the cloned genes (63%) were differentially regulated (in equal proportions from differential display reverse transcription-polymerase chain reaction and SHMB procedures), and of these, 24 (32%) had similarity to known genes or putative proteins (more from SHMB). The analysis provided insights into the signaling and transcriptional basis of direct and indirect defenses used against herbivores, suggesting simultaneous activation of salicylic acid-, ethylene-, cytokinin-, WRKY-, MYB-, and oxylipin-signaling pathways and implicating terpenoid-, pathogen-, and cell wall-related transcripts in defense responses. These defense responses require resources that could be made available by decreases in four photosynthetic-related transcripts, increases in transcripts associated with protein and nucleotide turnover, and increases in transcripts associated with carbohydrate metabolism. This putative up-regulation of defense-associated and down-regulation of growth-associated transcripts occur against a backdrop of altered transcripts for RNA-binding proteins, putative ATP/ADP translocators, chaperonins, histones, and water channel proteins, responses consistent with a major metabolic reconfiguration that underscores the complexity of response to herbivore attack.