Study On Mature Embryo Callus Tissue Cultivation And Seed Protein Composition Of Maize9417

In this article, we have selected photoperiod and thermo-sensitive maize9417maintainerlines mature embryos as the explants, with LS, MS, N6as the inducing culture medium ofcallus, and designing time gradient of explant pretreatment, to screen out a suitable mediumand the best pretreatment time of explants for inducing callus of maize mature embryo;through adding different concentration phytohormone and antagonists to research the controlof maize mature embryos inducement and subculture browning. Then four different genotypesof maize mature embrytos-9417maintainer lines, sterile lines, H: II and Zhengdan958whichused as explants were studied the effect of genotype on callus induction. In addition, the vigorof maize9417maintainer line seeds and sterile line seeds as the experimental material, andtreated by artificial aging (controlled deterioration test, CDT) of maintainer line seeds wasstudied that it is influenced germination of maize seed and changes of physiological activityand so on. Meanwhile, we take sterile line and maintainer line maize seeds as materials tostudy the changes of some relative physiological activity and the germination.We alsoconsider the endosperm and embryo proteome changes and peptide mass finferprinting of twogenotypes maize seeds in different treated, which we discuss the mechanisms of seed agingand repairing to provide a theoretical basis for storage security of germplasm resources.The results were as follows:1. Browning is most likely to occur in maize mature embryo callus, so solve the problemis important. The study discover that when the pretreatment time is36h on mature embryoexplants of maize callus, there has the most appropriate growth, callus texture close,thedifferentiation phenomenon is relatively diminished; Object to different culture mediuminfluenced on the rate of callus induction, found in the MS medium for callus growth the best,and browning degree is lower than the other two kinds of induced medium.2. Taking MS as basic culture medium, by studying effect of the mature embryos callsinduction on adding different concentrations of6-BA and2,4-D, the result showed, low andhigh concentration of6-BA and2,4-D all was not suitable to callus induction, the callusinduction rate was the highest when the concentration of6-BA was0.2mg/L and2,4-D was2.0mg/L, but the browning rate is relatively higher; by adding different concentrations ofAgNO3and mannitol in subculture medium, and supplemented6-BA and2,4-D on callus subcultures of mature embryo browning control effects. In the medium containing AgNO3,6-BA,2,4-D concentrations were6mg/L,0.2mg/L,2.0mg/L, the callus during subculturebrowning rate is lowest, and found when mannitol,6-BA,2,4-D concentrations were20g/L,0.2mg/L,2.0mg/L, compared with other concentration, Callus browning rate was the lowest,analysis showed significant difference.3. Through four genotypes of mature embryos to induce callus in the induction medium,different genotypes effects on callus induction, found that the difficult induction callus ofphotoperiod and thermo-sensitive maize9417mature embryo. So it is very difficult thatresearch the callus induction of mature embryos, subculture and differentiation futher. Atpresent, it is only a preliminary study about the induction and subculture of callus, andprovides a probability reference for research the establishment of regeneration system ofmaize9417mature embryo.4. To deal with the artificial aging of the maize maintainer lines seeds and made thegermination experiments. The results of germination experiments showed that differentaging time have influence on germination rate. With the increasing of the aging time,germination rate, germination potential, germination index had a dropping tendency. Thegermination rate dropped significantly after aged2days, the seed vigor of line was lostseriously, germination rate reached to0after7days artificial aged treatment. Besides,through the germination expermination of the sterile lines and maintainer lines untreatedseed that we have found the maintainer line seeds germination rate much higher than thesterile line seeds.5. The different aged treatment of the maintainer line maize seeds, we found the indexsare changed by measure the seed physiological index of different aged treatment times atsome degree. The results shows that soluble sugar content, soluble protein content, andenzymatic activity of SOD, POD, CAT gradually decreased tendency, while the content ofMDA increased with prolonging of the aging. Furthermore, through untreated seeds of themaintainer and sterile line researched, we obtain a result that the POD activity and SODactivity in sterile lines was higher than in maintainer lines, but the content of soluble sugar,the content of soluble protein, the content of MDA and CAT activity in sterile lines was lowerthan in maintainer lines.6. SDS-PAGE showed that the different aging treatment of maintainer line seedsendosperm protein and embryo protein were not different bands, or the content of protein is aminor change in embryos. To electrophoretic analyses found that the four different bandsbetween the sterile and maintainer line untreated maize seeds of embryo protein. With masspectrometry and bioinformatics analysis, obtain the possible six proteins: the Globulin-1S allele, the Globulin-2, the uncharacterized protein, Chaperonin CPN60-2, mitochondrial, andElongation factor Tu, the Glucose-1-phosphate adenylyltransferase. For analysis the functionof each protein, obtains the relationship between the difference proteins and sterile lines.