Abstract
BackgroundThioredoxin h (trx h) is closely related to germination of cereal seeds. The cDNA sequences of the thioredoxin s (trx s) gene from Phalaris coerulescens and the thioredoxin h (trx h) gene from wheat are highly homologous, and their expression products have similar biological functions. Transgenic wheat had been formed after the antisense trx s was transferred into wheat, and it had been certified that the expression of trx h decreased in transgenic wheat, and transgenic wheat has high resistance to pre-harvest sprouting.Methodology/Principal FindingsThrough analyzing the differential proteome of wheat seeds between transgenic wheat and wild type wheat, the mechanism of transgenic wheat seeds having high resistance to pre-harvest sprouting was studied in the present work. There were 36 differential proteins which had been identified by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF-MS). All these differential proteins are involved in regulation of carbohydrates, esters, nucleic acid, proteins and energy metabolism, and biological stress. The quantitative real time PCR results of some differential proteins, such as trx h, heat shock protein 70, α-amylase, β-amylase, glucose-6-phosphate isomerase, 14-3-3 protein, S3-RNase, glyceraldehyde-3-phosphate dehydrogenase, and WRKY transcription factor 6, represented good correlation between transcripts and proteins. The biological functions of many differential proteins are consistent with the proposed role of trx h in wheat seeds.Conclusions/SignificanceA possible model for the role of trx h in wheat seeds germination was proposed in this paper. These results will not only play an important role in clarifying the mechanism that transgenic wheat has high resistance to pre-harvest sprouting, but also provide further evidence for the role of trx h in germination of wheat seeds.
Highlights
Pre-harvest sprouting (PHS) is a major limitation to stability of wheat production in parts of the world where long range rainfall or damp conditions prior to harvest often occur
In order to investigate the changes of wheat seed proteome in response to antisense trx s gene, 2-DE analysis of the total proteins in wild type and transgenic wheat seeds in different stages was carried out
Spot intensity variations between wild type and transgenic samples were quantified by software image analysis, and the protein spots showing significant differences (p,0.05) were selected. 20 proteins were selected as ‘‘significantly differential proteins’’ in Figure 1, including 12 down-regulated and 8 up-regulated proteins in transgenic wheat seeds. 16 proteins were considered as ‘‘significantly differential proteins’’ in Figure 2, including 9 downregulated and 7 up-regulated proteins in transgenic imbibed wheat seeds
Summary
Pre-harvest sprouting (PHS) is a major limitation to stability of wheat production in parts of the world where long range rainfall or damp conditions prior to harvest often occur. To clarify the mechanism of transgenic wheat seeds having high resistance to PHS, the changes in protein profile of wheat seeds were identified by comparative proteomics between wild type and transgenic wheat seeds. The abundant changes in these identified proteins, as well as their putative functions, are consistent with the proposed role of trx h in wheat seeds. This is the first proteomic study of the wheat seeds in response to trx h, and the results greatly expand our knowledge about the role of thioredoxin h in germination of wheat seeds. A possible model for the role of trx h in wheat seeds germination was proposed in this paper
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