Double overexpression of DREB and PIF transcription factors improves drought stress tolerance and cell elongation in transgenic plants

Mungbean (Vigna radiata L.) is a valuable legume crop grown in tropical and subtropical areas of Asia. Drought is one of the major factors hindering its growth globally. APETALA2/ethylene-responsive element factor binding proteins (AP2/ERF) are an important family of plant-specific transcription factors (TFs) involved in drought-stress tolerance. We identified 71 AP2/ERF TFs in the mungbean genome by using bioinformatics tools and classified them into subfamilies: AP2 (16 members), ERF (22), RAV (2), DREB (30) and soloist (other proteins with no domain, 1). Members of DREB play a critical role in drought-stress tolerance. Ten-day-old mungbean plants cv. AZRI-06 were exposed to drought stress by complete withholding of water for 7 days. Root samples were collected from control and drought-stressed plants, and the expression pattern of 30 identified VrDREB genes was determined by qPCR. Most VrDREB genes exhibited differential expression in response to drought. Five genes (VrDREB5, VrDREB12, VrDREB13, VrDREB22, VrDREB30) were highly expressed under drought stress and might be considered excellent candidates for further functional analysis and for improvement of mungbean drought tolerance.

Download Full-text

Overexpression of AtAHL20 causes delayed flowering in Arabidopsis via repression of FT expression

10.21203/rs.3.rs-38575/v1 ◽

2020 ◽

Author(s):

Reuben Tayengwa ◽

Pushpa Sharma-Koirala ◽

Courtney F. Pierce ◽

Breanna E Werner ◽

Michael M Neff

Keyword(s):

Drought Stress ◽

Transgenic Plants ◽

Gene Family ◽

Stress Tolerance ◽

Flowering Time ◽

Camelina Sativa ◽

Negative Regulator ◽

Wild Type ◽

Late Flowering ◽

Drought Stress Tolerance

Abstract Background The 29-member Arabidopsis AHL gene family is classified into three main classes based on nucleotide and protein sequence evolutionary differences. These differences include the presence or absence of introns, type and/or number of conserved AT-hook and PPC domains. AHL gene family members are divided into two phylogenetic clades, Clade-A and Clade-B. A majority of the 29 members remain functionally uncharacterized. Furthermore, the biological significance of the DNA and peptide sequence diversity, observed in the conserved motifs and domains found in the different AHL types, is a subject area that remains largely unexplored. Results Transgenic plants overexpressing AtAHL20 flowered later than the wild type. Transcript accumulation analyses showed that 35S:AtAHL20 plants contained reduced FT, TSF, AGL8 and SPL3 mRNA levels. Similarly, overexpression of AtAHL20’s orthologue in Camelina sativa, Arabidopsis’ closely related Brassicaceae family member species, conferred a late-flowering phenotype via suppression of CsFT expression. In addition, 35S:AtAHL20 seedlings exhibited suppressed hypocotyl length and enhanced water stress tolerance. However, overexpression of an aberrant AtAHL20 gene harboring a missense mutation in the AT-hook domain’s highly conserved R-G-R core motif abolished the late-flowering phenotype. Data from targeted yeast-two-hybrid assays showed that AtAHL20 interacted with itself and several other Clade-A Type-I AHLs which have been previously implicated in flowering-time regulation: AtAHL22, AtAHL27 and AtAHL29. Conclusion We showed via gain-function analysis that AtAHL20 is a negative regulator of FT expression, as well as other downstream flowering time regulating genes. A similar outcome in Camelina sativa transgenic plants overexpressing CsAHL20 suggest that this is a conserved function. Additionally, overexpression of AtAHL20 resulted in shorter hypocotyls and enhanced drought stress tolerance compared to wild-type plants. Our results demonstrate that AtAHL20 is a negative regulator of transition to flowering and hypocotyl elongation, but a positive regulator of drought stress tolerance.

Download Full-text