Giri Ashok P, Wünsche Hendrik, Mitra Sirsha, Zavala Jorge A, Muck Alexander, Svatos Ales, Baldwin Ian T
Department of Molecular Ecology, Max Planck Institute for Chemical Ecology, 07745 Jena, Germany.
Plant Physiol. 2006 Dec;142(4):1621-41. doi: 10.1104/pp.106.088781. Epub 2006 Oct 6.
When Manduca sexta attacks Nicotiana attenuata, fatty acid-amino acid conjugates (FACs) in the larvae's oral secretions (OS) are introduced into feeding wounds. These FACs trigger a transcriptional response that is similar to the response induced by insect damage. Using two-dimensional gel electrophoresis, matrix-assisted laser desorption ionization-time of flight, and liquid chromatography-tandem mass spectrometry, we characterized the proteins in phenolic extracts and in a nuclear fraction of leaves elicited by larval attack, and/or in leaves wounded and treated with OS, FAC-free OS, and synthetic FACs. Phenolic extracts yielded approximately 600 protein spots, many of which were altered by elicitation, whereas nuclear protein fractions yielded approximately 100 spots, most of which were unchanged by elicitation. Reproducible elicitor-induced changes in 90 spots were characterized. In general, proteins that increased were involved in primary metabolism, defense, and transcriptional and translational regulation; those that decreased were involved in photosynthesis. Like the transcriptional defense responses, proteomic changes were strongly elicited by the FACs in OS. A semiquantitative reverse transcription-PCR approach based on peptide sequences was used to compare transcript and protein accumulation patterns for 17 candidate proteins. In six cases the patterns of elicited transcript accumulation were consistent with those of elicited protein accumulation. Functional analysis of one of the identified proteins involved in photosynthesis, RuBPCase activase, was accomplished by virus-induced gene silencing. Plants with decreased levels of RuBPCase activase protein had reduced photosynthetic rates and RuBPCase activity, and less biomass, responses consistent with those of herbivore-attacked plants. We conclude that the response of the plant's proteome to herbivore elicitation is complex, and integrated transcriptome-proteome-metabolome analysis is required to fully understand this ubiquitous ecological interaction.
当烟草天蛾攻击黄花烟草时，幼虫口腔分泌物（OS）中的脂肪酸 - 氨基酸共轭物（FACs）会被引入取食伤口。这些FACs引发一种转录反应，该反应类似于昆虫损伤诱导的反应。我们使用二维凝胶电泳、基质辅助激光解吸电离飞行时间质谱和液相色谱 - 串联质谱，对幼虫攻击诱导的酚类提取物和叶片细胞核组分中的蛋白质进行了表征，以及/或者对受伤并分别用OS、无FAC的OS和合成FAC处理的叶片中的蛋白质进行了表征。酚类提取物产生了大约600个蛋白点，其中许多因诱导而发生变化，而细胞核蛋白组分产生了大约100个蛋白点，其中大多数不受诱导影响。对90个可重复的诱导剂诱导变化的蛋白点进行了表征。一般来说，增加的蛋白质参与初级代谢、防御以及转录和翻译调控；减少的蛋白质参与光合作用。与转录防御反应一样，蛋白质组的变化也被OS中的FACs强烈诱导。基于肽序列的半定量逆转录 - PCR方法用于比较17种候选蛋白质的转录本和蛋白质积累模式。在6种情况下，诱导的转录本积累模式与诱导的蛋白质积累模式一致。通过病毒诱导的基因沉默对鉴定出的一种参与光合作用的蛋白质——核酮糖-1,5-二磷酸羧化酶激活酶进行了功能分析。核酮糖-1,5-二磷酸羧化酶激活酶蛋白水平降低的植物光合速率和核酮糖-1,5-二磷酸羧化酶活性降低，生物量减少，这些反应与受食草动物攻击的植物一致。我们得出结论，植物蛋白质组对食草动物诱导的反应是复杂的，需要综合转录组 - 蛋白质组 - 代谢组分析才能充分理解这种普遍存在的生态相互作用。
