Study Of Sunflowers Seedling(Helianthus Annuus L.) Under Lead Stress About Physiology And Proteomics

For the study of sunflower seedlings with lead stress after a series of dynamic change and the relationship between protein structure and protein function, this study sunflower(Helianthus annuusL.) varieties HA89 for material, in the concentration of Pb + 0(control), 200 mg/L, 400 mg/L, 600 mg/L, 800 mg/L lead acetate solution treatment, a total of 30 days. By determining germination rate, germination potential and growth index and catalase, peroxidase and other physiological index to analyze the sunflower seedlings under different levels of lead stress changes or after different degrees of damage; By combining infrared spectrum and two-dimensional electrophoresis technique to study the sunflower seedlings of lead stress after the dynamic change of protein secondary structure and the protein group, in order to clearly proteomics and the change of protein structure, this study chooses 30 days lead concentration is 0(control), 400 mg/L, 800 mg/L for protein secondary structure and the protein expression of research;In addition, in order to provide suitable sunflower seedling two-dimensional electrophoresis system, this study used different methods to extract protein and focus on different time to further optimize the sunflower seedling of two-dimensional electrophoresis system.The results showed that:From the result of the growth index, low concentration of Pb+ stress treatment of sunflower seedling germination rate, germination rate, vigor index and seedling weight, more than a certain concentration of high concentrations of lead stress on germination rate, germination rate, vigor index and seedling weight inhibition; And germination index of lead stress influence is not obvious, root length and seedling length declined with the increase of lead stress concentration, root decreased with the increase of lead stress concentration on the rise trend.From physiological indexes of learn that with the increase of lead stress concentration, MDA content in sunflower seedling performance after the first drop in the rising trend, SOD, POD and CAT activity were first after rising downward trend.From Fourier infrared spectrum testing different sunflower seedlings after lead stress caused by the changes of protein secondary structure, sunflower seedling root beta folding structure with lead stress concentration increases, the number of sunflower seedlings treated by different concentration of lead stress of stem and leaf contains β-folding structure ‘s number of change is more obvious, so sunflower seedling root, stem and leaf of protein secondary structure in beta- fold structure is most sensitive to lead stress;Sunflower seedling roots with the increase of lead stress concentration, protein secondary structure more stable, and the department of stem and leaf with the increase of lead stress concentration, protein secondary structure by a certain degree of damage, tend to be loose.In establishing a two-dimensional electrophoresis system was made on the results, the Tris phenol extraction, TCA- acetone extraction and water extraction process of the three methods to extract the sunflower seedlings protein contains two, all of them, and using Tris phenol extraction method to extract the total protein content of the higher relative to other two methods; Always focus on six hours in two-dimensional electrophoresis, contrast and 10 hours of the two methods to study the sunflower seedling system of two-dimensional electrophoresis, the results show that focus on six hours of system is more suitable for the sunflower seedlings.From the two-dimensional electrophoresis and mass spectrometry after testing different lead stress change of protein expression result of sunflower seedlings, after lead stress treatment were lower than the number of sunflower seedlings protein points without number of sunflower seedlings protein point lead stress treatment.After analysis found that differences in protein by ImagemasterTM2 D Platinum software point total of 27, significant difference of protein point(that is, the difference in multiples of 2 times more) 11, mass spectrum successfully identified nine of them;After mass spectrometry analysis and compare protein libraries that differences of protein, such as chloroplast protein, induced protein, are all with sunflower response to heavy metal stress reaction mechanism of important proteins.