Alfalfa is widely grown and is one of the most important forage crops in the world, but its growth and biomass production are markedly reduced under salt and drought stress, particularly during the early seedling stage. To identify the mechanisms behind salt and drought responsiveness at the alfalfa seedling stage, the proteins expressed were analyzed under no-treatment, 200 mol L^–1 NaCl and 180 g L^–1 PEG treatment conditions during the seedling stage. Out of more than 800 protein spots detected on two-dimensional electrophoresis (2-DE) gels, 35 proteins showed statistically significant responses (P<0.05) to NaCl and PEG stress, which were selected for tandem mass spectrometric identification, owing to their good resolution and abundance levels, and 32 proteins were positively identified. The identified proteins were divided into seven functional categories: photosynthetic metabolism, protein biosynthesis, folding and assembly, carbohydrate metabolism-associated proteins, stress defense related protein, metabolism of nucleic acid, other function categories and unknown proteins. Our results suggested that these proteins may play roles in alfalfa adaptation to salt and drought stress. Further study of these proteins will provide insights into the molecular mechanisms of abiotic stress and the discovery of new candidate markers in alfalfa.

This study was designed to determine the effect of different sperm preparation treatments on yak sperm quality and in vitro embryo development. Frozen-thawed semen samples were treated using swim-up or percoll gradient centrifugation methods. Sperm concentration, progressive motility, recovery of motile sperm, membrane integrity, acrosome and chromatin integrity were scored and compared in recovered samples and controls. In addition, the effects of two sperm separation treatments on embryos capable of cleavage and in vitro development to the blastocyst stage were evaluated. Swim-up separated sperm showed a higher motility, while the concentration of spermatozoa recovered and percent recovery of motile sperm were higher with percoll gradient centrifugation separation. According to the optical and electron microscopies, swim-up produced higher percentage of sperm with intact plasma membrane and acrosome than percoll gradient centrifugation separation. However, there was no difference in the percentage of sperm with intact chromatin between two treatment groups. Cell numbers in the blastocysts of two groups were not different. The blastocyst rate was similar in both groups, whereas cleavage rate was higher when swim-up was used.