Abstract
Once candidate genes are available, the application of genetic transformation plays a major part to study their function in plants for adaptation to respective environmental stresses, including waterlogging (WL). The introduction of stress-inducible genes into wheat remains difficult because of low transformation and plant regeneration efficiencies and expression variability and instability. Earlier, we found two cDNAs encoding WL stress-responsive wheat pathogenesis-related proteins 1.2 (TaBWPR-1.2), TaBWPR-1.2#2 and TaBWPR-1.2#13. Using microprojectile bombardment, both cDNAs were introduced into “Bobwhite”. Despite low transformation efficiency, four independent T2 homozygous lines for each gene were isolated, where transgenes were ubiquitously and variously expressed. The highest transgene expression was obtained in Ubi:TaBWPR-1.2#2 L#11a and Ubi:TaBWPR-1.2#13 L#4a. Using quantitative proteomics, the root proteins of L#11a were analyzed to explore possible physiological pathways regulated by TaBWPR-1.2 under normal and waterlogged conditions. In L#11a, the abundance of proteasome subunit alpha type-3 decreased under normal conditions, whereas that of ferredoxin precursor and elongation factor-2 increased under waterlogged conditions in comparison with normal plants. Proteomic results suggest that L#11a is one of the engineered wheat plants where TaBWPR-1.2#2 is most probably involved in proteolysis, protein synthesis and alteration in the energy pathway in root tissues via the above proteins in order to gain metabolic adjustment to WL.
Highlights
In the last two decades, genetic transformation has become a powerful tool to transfer new genes into crop plants
In a previous study on morphological adaptation to WL in the seminal roots of hexaploid spring wheat “Bobwhite SH 98 26” [25], we found that levels of a TaPR-1.2 significantly increased during lysigenous aerenchyma formation [26]
We analyzed the expression of transgenes in various organs of four homozygous lines and one null-segregant from ubiquitin promoter and intron (Ubi):TaBWPR-1.2#13 in the absence of stress
Summary
In the last two decades, genetic transformation has become a powerful tool to transfer new genes into crop plants. Promising stress-inducible promoters, such as Arabidopsis rd29A [8], maize Rab17 [6] and barley HvDhn4s [7], have been used to study the effect of drought, but these promoters may be not efficient in heterologous systems. To circumvent these problems, some wheat genes, stress-inducible ones, have been overexpressed in other plants [14,15]. Little is known about PR-1.2 gene expression and protein production in root in response to environmental stresses, relevant information has been obtained for other PR families. This work may be useful for those who attempt to produce transgenic wheat plants and for those interested in the role of PR-1.2 proteins in wheat
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