Proteomics combined with BSMV-VIGS methods identified some N deficiency-responsive protein species and ABA role in wheat seedling
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Crops often encounter a soil deficiency of nitrogen (N), the most important macronutrient for plants; however, the molecular mechanism of plant responses to N deficiency remains unclear. In this study, proteome-level changes that occur in bread wheat seedlings suffering from N deficiency were investigated to identify some N deficiency-responsive protein species in bread wheat.
We utilized isobaric tagging for relative and absolute quantification (iTRAQ) to measure changes in the proteome patterns of N-deficient wheat seedlings and validated the role of abscisic acid (ABA) using the barley stripe mosaic virus-induced gene-silencing (BSMV-VIGS) method.
A total of 1515 N deficiency–responsive protein species were successfully identified in both root and leaf tissues of wheat seedlings suffering from 8-d N deficiency. Of these, abundance of wheat zeaxanthin epoxidase (TaZEP), a key ABA synthesis-related enzyme, was significantly upregulated, and the endogenous ABA contents also markedly increased. After TaZEP gene was further silenced using BSMV-VIGS method, BSMV-VIGS-TaZEP infected wheat seedlings showed enhanced sensitivity to N deficiency, suggesting silencing of TaZEP gene decreased the tolerance to N deficiency remarkably.
Our results identified some N deficiency-responsive protein species and revealed the role of ABA in wheat responses to N deficiency.
KeywordsAbscisic acid BSMV-VIGS iTRAQ Nitrogen deficiency Proteome Triticum aestivum L
barley stripe mosaic virus-virus induced gene-silencing
isobaric tagging for relative and absolute quantification
quantitative real-time PCR
This study was financially supported by the Science and Technology Innovation Program for Increase in Yield and Efficiency of Food Crop (2016YFD0300105) and the National Key Technology Support Program (2015BAD26B01).
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