Diferential Proteomics Of Leaf From Tobacco Varieties With Different Pressure Resistance

Nicotiana tabacum L. is one of China,s important economic crops, which play an important rolein national economy. Sustainable Development of tobacco production is restricted by drought stress andfungi disease caused by Phytophthora nicotianae Breda de Haan.Molecular mechanism of tobacco geneexpression and Specific protein synthesis induced by drought sress and Phytophthora parasitica var.nicotianae remains to be elucidated in detail. In this study, Comparative proteomics, especiallytwo-dimensional electrophoresis (2-DE), was carried out to investigate the change of tobacco leafproteome from 4 tabacco varieties with different resistance after the treatment of drought orPhytophthora parasitica var. nicotianae inoculation. The main results are as follows.1 Experiment conditions are optimized preliminarily, such as Total protein extract methods oftobacco leaf, immobilized pH gradient, protein quantity, and so on.2 Zhongyan 14,a drought-tolerant variety,transplanted for 45 days,was stressed drought for 12days. we recognized more than 300 protein spots on 2-DE profiles,from which 52 differentiallyexpressed protein spots was screened out. fourteen protein spots were up-regulated,eighteen weredown-regulated, fifteen and no five were detected only in the gels of the treated plants.By tandem MSand multifarious database search methods, 42 protein spots (19 proteins) were successfully identified,Including 31 protein spots (9 proteins) related to material and energy metabolism, 4 antioxidativerelated protein spots (4 proteins), 3 protein spots (3 proteins) participated in translation, 2 protein spots(2 proteins) participated in signal transduction, 2 ungrouped protein spots.3 Zhongyan 100,a drought-sensitive variety,transplanted for 45 days,was stressed drought for 12days. we recognized more than 330 protein spots on 2-DE profiles,from which 43 differentiallyexpressed protein spots was screened out. twenty protein spots were up-regulated,ten weredown-regulated, twelve and no one were detected only in the gels of the treated plants.By tandem MSand multifarious database search methods, 34 protein spots (14 proteins) were successfully identified,Including 28 protein spots (8 proteins) related to material and energy metabolism, 4 antioxidativerelated protein spots (4 proteins), 1 protein spots (1 proteins) participated in signal transduction, 1ungrouped protein spots.4 Xiaohuangjin 1025,a variety susceptible for the tobacco black shank pathogen,3 weeks aftertemporary planting,was inoculated with Phytophthora parasitica var. nicotianae spore suspension for 3days. we recognized 260 or so protein spots on 2-DE profiles, from which 38 differentially expressedprotein spots was screened out. Fourteen protein spots were up-regulated,sixteen were down-regulated,six and no two were detected only in the gels of the treated plants. By tandem MS and multifariousdatabase search methods, 29 protein spots (15 proteins) were successfully identified, Including 20protein spots (9 proteins) related to material and energy metabolism, 1 antioxidative related proteinspots (1 proteins), 4 protein spots (3 proteins) participated in translation,1 protein spots (1 proteins)participated in signal transduction, 3 ungrouped protein spots(2 proteins). 5 Gexinsanhao,a variety of resistance to the tabacco black shank pathogen,3 weeks after temporaryplanting,was inoculated with Phytophthora parasitica var. nicotianae spore suspension for 3 days. werecognized 400 or so protein spots on 2-DE profiles, from which 39 differentially expressed proteinspots was screened out.Fourteen protein spots were up-regulated,twenty were down-regulated, threeand no two were detected only in the gels of the treated plants. By tandem MS and multifariousdatabase search methods, 28 protein spots (19 proteins) were successfully identified, Including 15protein spots (8 proteins) related to material and energy metabolism, 8 antioxidative related proteinspots (7 proteins), 1 protein spots (1 proteins) participated in signal transduction, 2 protein spots (2proteins) related to defence, 2 ungrouped protein spots(2 proteins).Based on the above results,we can definite that most of identified proteins involved in energy andmaterial metabolism．In addition,many defense proteins were also among them．We discussed themechanism of tobacco response to drought and Phytophthora parasitica var. nicotianae stressesprimarily, shed light on the relationship between environmental stresses and tobacco,s resistancecapability. This study set a favorable theoretical basis for further selection of specific proteins relatedresistance.These specific proteins can serve as a mark to tobacco resisitace to breeding.