Seed Proteomic Analysis Of TrxS Transferred Barley Lines

The content of protein in malting barley's grains and its velocity of metabolisation in germination are crucial to assess the malts'brewing quality. In this paper,firstly,malting barley seeds of several TrxS gene transferred lines (Franklin and Harrington strains,expressed by F and H) were used to analysis the changes of storage protein fractions in the endosperm before and after its germination by sequential extraction,and to find differentially expressed proteins by SDS-PAGE,which aimed to explore the effection of TrxS gene on the rate of barley germination from the proteomics point of view amd to evaluate its feasibility on the malts'quality improvement; Secondly, the TrxS gene transferred dwarf mutant lines,H-7-5-14-2 and H-7-5-14-5,whose agronomic traits are dwarf and anti-lodging steadily were used to analysis its plant height at 15dpa by statistical methods,and then,H-7-5-14-2,one of these two dwarf mutant lines was used to analysis its protein profiles of grains and seedling (trefoil stage) leaves by 2-DE,in which the traditional TCA-Ac methods was used to extract their total proteins. The results showed that:1. The determination of protein content in raw and germinated seeds showed that: there is no significant between the transgenic and non-transgenic lines on total protein content in the raw endosperm of all tested materials,nor on any kinds of protein fractions; When germinated at 2ds,the storage proteins in the endosperm of most transgenic strains(but any ones),which include total protein, protein fractions of globulin, hordein and glutelin,are decomposited rapidly and highly significanted compared to its control; In contrast,the content of albumin is raised, which is highly significanted too between the transgenic and non-transgenic strains.2. The SDS-PAGE profiles of the protein fractions in the raw and germinated endosperm of tested seeds showed that:(1) In the albumin profiles,the number of highly molecular weight strips is decreased and the lowly molecular weight strips are increased when the grain germinated at 2ds; (2) In the globulin profiles,there is a strip that expressed obviously different both at 66.2-94.0kD and 35.0-45.0 kD in the raw grains of F strains; There is a degraded strip both lower at 66.2 kD and upper at 45.0 kD in the H-CK malts which are germinated at 2ds compared to the raw grains,while it is absent in the transgenic lines; (3) In the hordein profiles,a unique expressed strip appeared at 35.0-45.0 kD both in F and H-CK strains compared to their transgenic lines,and it was stability when the grain germinated at 2ds; (4) In the glutelin profiles,there is no different expressed strips detected among all of the strains,and the number of strips separated in every lanes is limited.3. Statistical analysis showed that,the mean plant height of the two semi-dwarf strains was 17.25cm and 16.63cm lower than H-CK,respectively,and both of which were highly significanted. Furthermore,the 2-DE protein profiles of grains and seedling (trefoil stage) leaves showed that,there is many protein spots that expressed significantly between H-7-5-14-2 and H-CK,which is non-transgenic lines,and some ones expressed highly significanted.