Effects Of Exogenous Snp On Physiological,biochemical And Differential Proteins In Sun-flower Seedlings Under Cadmium Stress

The research material was sunflower HA300 inbred seedlings.Firstly,orthogonal test L9(33)was used,and different concentrations of Cd Cl2,sodium nitroprusside(SNP)and different culture days were used as three factors.Firstly,the antioxidant enzyme activities(SOD,POD,CAT)and malondialdehyde(MDA)content of 9 groups were measured to investigate the treatment of sunflower seedlings with the least treatment in 9 groups.Secondly,9 groups were treated.The content of metal after treatment was measured,and the results of biochemical indicators were combined to obtain the most effective treatment method for sunflower seedlings to resist Cd toxicity.Finally,0 ?M Cd Cl2 stress,400 ?M Cd Cl2 stress and sunflower seedlings cultured under the most effective Cd toxicity were extracted.Whole protein,and its concentration,used for two-dimensional electrophoresis analysis,looking for differential proteins,and finally using MALDI-TOF-TOF-MS to analyze differential proteins,the main results are as follows:100 and 200 ?M Cd Cl2,50 and 100 ?M SNP were added respectively,even if the culture days were different,the SOD activity was similar.After treatment with 9 groups,under the stress of 400 ?M Cd Cl2,the activity of SOD was the highest after added 50 ?M SNP for 8 days,reaching 157.85 U·g-1FW;the activity of POD was higher in groups 3,5 and 7.The third group treatment was 1.36 times and 3.24 times the activity of the first and second groups,while the seventh group treatment was 1.60 times of the third group treatment,and the POD activity was as high as 253.75 U·g-1FW;the CAT activity was under the stress of 100 ?M Cd Cl2.Different concentrations of SNP were added,and the variation range was smaller than that of the other treatment groups.Under the stress of 400 ?M Cd Cl2,the activity of the three groups decreased,and the activity of added 50?M SNP for 8 days was higher than that of the other treatment groups;MDA content Under the stress of 200 ?M Cd Cl2,the change was not obvious,but the change was obvious under the treatment of the second and seventh groups.The content of the second group was the highest,and the content of the seventh group was the lowest.The above preliminary indicates that under the stress of 400 ?M Cd Cl2,added 50 ?M SNP for 8 days can make sunflower seedlings effectively resist Cd toxicity.The absorption rate of Cd in sunflower seedlings treated with 9 groups was lower in Groups 2,6 and 7 and decreased in turn.Under the stress of 100 ?M Cd Cl2,added 100 ?M,200 ?M SNP and 200 ?M Cd Cl2,added 100 ?M SNP and 400 ?M Cd Cl2,added 50 ?M SNP,the total amount of Mg was approximately the same for different days.But the total amount of Mg was the highest when the medium was added with 400 ?M Cd Cl2 stress,50 ?M SNP for 8 days.The change of total Fe content was the most obvious and the highest content was treated in the 9th group,which is consistent with the total K amount.The total Ca content was higher in the first,fifth and seventh groups,which was higher than that of the other treatment groups,and seventh groups was higher than that in the other treatment group.The total Zn amount was significantly higher than that in the seventh group.The other groups change were smaller and lower than the seventh group treatment;the total Na amount was higher in the sixth and seventh groups,and the total Na amount in the sixth group was more than double that in the other seven groups,and the seventh group was treated.It is 1.55 times of the sixth group.In summary,under the stress of 400 ?M Cd Cl2,added 50 ?M SNP for 8 days can alleviate the Cd toxicity of sunflower seedlings and make it grow normally.The whole protein of sunflower seedlings cultured with 0 ?M Cd Cl2 stress,400 ?M Cd Cl2 stress and 400 ?M Cd Cl2 added 50 ?M SNP was extracted,and the total protein content was 42.171 ?g??L-1,35.983 ?g??L-1 39.421 ?g??L-1 by UV absorption A280 method.Three groups of whole proteins were used for two-dimensional electrophoresis analysis.The analysis of ImagemasterTM2 D Platinum software showed that the change of protein spots was consistent with the change of whole protein content.The most stress was 0 ?M Cd Cl2,which was 217,followed by the addition of 50 ?M SNP protein spots and 400 ?M Cd Cl2 stress.Which protein spots were the least.And there were 11 differential protein spots with a magnification of 4 times or more.Using MALDI-TOF-TOF-MS to identify the information and function of 11 differential protein spots,a total of 8 were successfully identified.According to their functions,they can be divided into 5 categories,which control plant growth and traits of legumin;Triose phosphate isomerase and 2,3-bisphosphoglycerate-independent phosphoglycerate mutase;aldehyde/histamine dehydrogenase,L-ascorbate: peroxidation associated with oxidative decomposition of plants Hydroxide reductase and superoxide dismutase(Cu-Zn);protein inhibitors of Kunitz-like protein that regulate enzyme activity;class B acid phosphatase associated with plant phosphorus deficiency.