An aquaporin protein is associated with drought stress tolerance.

Water channel proteins known as aquaporins (AQPs) regulate the movement of water and other small molecules across plant vacuolar and plasma membranes; they are associated with plant tolerance of biotic and abiotic stresses. In this study, a PIP type AQPs gene, designated as GoPIP1, was cloned from Galega orientalis, a high value leguminous forage crop. The GoPIP1 gene consists of an 870 bp open reading frame encoding a protein of 289 amino acids, and belongs to the PIP1 subgroup of the PIP subfamily. The transcript level of GoPIP1 was higher in the root of G. orientalis than in the leaf and stem. The level of GoPIP1 transcript increased significantly when treated with 200 mM NaCl or 20% polyethylene glycol (PEG) 6000. Transient expression of GoPIP1 in onion epidermal cells revealed that the GoPIP1 protein was localized to the plasma membrane. Over-expression of GoPIP1 increased the rosette/root ratio and increased sensitivity to drought in transgenic Arabidopsis plants. However, GoPIP1 over-expression in Arabidopsis had no significant effect under saline condition. The present data provides a gene resource that contributes to furthering our understanding of water channel protein and their application in plant stress tolerance.

Ectopic expression of tea MYB genes alter spatial flavonoid accumulation in alfalfa (Medicago sativa).

Flavonoids are one of the largest secondary metabolite groups, which are widely present in plants. Flavonoids include anthocyanins, proanthocyanidins, flavonols and isoflavones. In particular, proanthocyanidins possess beneficial effects for ruminant animals in preventing lethal pasture bloat. As a major legume forage, alfalfa (Medicago sativa) contains little proanthocyanidins in foliage to combat bloat. In an attempt to improve proanthocyanidin content in alfalfa foliage, we over-expressed two MYB transcription factors (CsMYB5-1 and CsMYB5-2) from tea plant that is rich in proanthocyanidins. We showed that, via targeted metabolite and transcript analyses, the transgenic alfalfa plants accumulated higher levels of flavonoids in stems/leaves than the control, in particular anthocyanins and proanthocyanidins. Over-expression of CsMYB5-1 and CsMYB5-2 induced the expression levels of genes involved in flavonoid pathway, especially anthocyanin/proanthocyanidin-specific pathway genes DFR, ANS and ANR in stems/leaves. Both anthocyanin/proanthocyanidin content and the expression levels of several genes were conversely decreased in flowers of the transgenic lines than in control. Our results indicated that CsMYB5-1 and CsMYB5-2 differently regulate anthocyanins/proanthocyanidins in stems/leaves and flowers. Our study provides a guide for increasing anthocyanin/proanthocyanidin accumulation in foliage of legume forage corps by genetic engineering. These results also suggest that it is feasible to cultivate new varieties for forage production to potentially solve pasture bloat, by introducing transcription factors from typical plants with high proanthocyanidin level.

Functional characterization of a gibberellin receptor and its application in alfalfa biomass improvement.

Bioactive gibberellins (GAs) are essential phytohormones involved in the regulation of many aspects of plant development. GA receptors are crucial in GA signal transduction in plants. The GA receptor GoGID1 promotes plant elongation and improves biomass production when ectopically expressed in tobacco. Here, we discovered that GoGID1 can interact with the DELLA proteins of Arabidopsis in the presence of gibberellic acid. GoGID1 partially or completely functionally rescued the phenotypes of the Arabidopsis double-mutants atgid1a/atgid1c and atgid1a/atgid1b. The overexpression of GoGID1 led to increases in plant height and biomass production in transgenic Arabidopsis plants. The GoGID1 gene enhanced GA sensitivity of the transgenic plants. More importantly, transgenic alfalfa plants overexpressing GoGID1 exhibited increased growth rates, heights and biomass and produced larger leaves when compared with the control plants. Thus, GoGID1 functions as a GA receptor, playing multiple roles in plant growth and development. The GoGID1 gene has the potential to be used in the genetic engineering of forage crops for biomass improvement.

Overexpression of the Galega orientalis gibberellin receptor improves biomass production in transgenic tobacco.

Gibberellins (GAs) are well-known phytohormones that contribute to a wide range of plant growth and development functions including stem elongation and leaf expansion. GA receptors perceive GA and transmit signals to activate GA-regulated reactions. In this study, a GA receptor gene with homology to other leguminous plants was isolated from Galega orientalis and termed GoGID. The 1732-bp full-length GoGID gene included an open reading frame of 1035 bp encoding a peptide of 344 amino acids. Sequence analysis indicated that GoGID shares conserved HGGS motif and active amino acid sites (Ser-Asp-Val/IIe) that are essential for maintaining it GA-binding activity. GoGID mRNA expression was more abundant in leaves than in roots or stems and could be up-regulated by the exogenous hormones. Overexpression of GoGID in transgenic tobacco plants promoted plant elongation and improved biomass production. These results suggested that GoGID functions as a GA receptor to alter GA-mediated signaling. GoGID may have a role in genetic engineering for the improvement of forage crops.