Proteomic Analysis Of The Chemical Hybridization Agents SX-1Induce Male Sterility Of Brassica Napus

Rapeseed (Brassica napus) is one of the main oilseed crops. The major task forbreeders is to increase the seed production at present. Hybrid cultivars have been used toincrease the production of rapeseed worldwide successfully. Male sterility of rapeseedinduced by chemical hybridization agents (CHA) is one of the main ways to producehybrid rapeseed. SX-1as a new-typed CHA had been widely applied to increase hybridseed production with high efficiency in China. A series of sterile lines that induced bySX-1, such as Y133、Sn338、YD66A had been selected out and new hybrids cultivars whichcontain Qinyou33, Qinyou19, Qinzayou5, Qinrong2and Qinzayou19were also cultivatedsuccessfully.As far as we known, the proteomic study on male sterile of anthers thatinduced by CHA in B. napus has not been reported yet.In order to observe the anthers changes after SX-1treatment in B. napus. Firstly, thedevelopment of microspore and tapetum were observed by using transmission electronmicroscopy (TEM) and scanning electron microscopy (SEM). The results show that, fewobvious morphological changes between microspores of control group and SX-1treatmentgroup in the beginning. All of them seemed to be full and round. However, nearly100%oftreated pollen grains were crimpled and these pollen coats lacked in the late stage ofpollen grains in maturation process. Plasmolysis occurred in the crimpy pollen grains. Theorganelles were not evident and the microspores were almost empty of contents at last.Meanwhile, in the early stage of anthers development, developing tapetosomes andelaioplasts could be observed both in SX-1treatment group and control group. However,tapetosomes and elaioplasts in tapetum treated with SX-1became disordered and brokenin advance in the late stage of pollen grains maturation process, resulting in tapetalextrusions and no oil body formation in treated pollen grains. These extruded regionsappeared to have an extremely high density of organelles and electron-dense materials.These results suggested that SX-1ruined the proper development of oil body in anthers.We also found that the filaments became shorter and thinner, the petals became smaller inthe treated group than in the control group.Proteomic analysis was conducted in different development stages of anthers after treated with6mg/L SX-1. Changes were analysized between SX-1treated and ultrapurewater treated anthers by using two-dimensional electrophoresis (2-DE). About1000protein spots were detected on each gel, a total of130,220,329,366protein spots weredown-regulated and87,25,74,60protein spots were up-regulated in response to SX-1treatment at four different developmental stages, Proteins were identified byliquid-chromatography–tandem mass spectrometry. A comprehensive analysis of the B.napus anthers proteome could be performed by using the available databases for rapeseedand other species.