Article

Molecular Biology Reports

, Volume 37, Issue 2, pp 745-753

First online:

Screening of genes induced by salt stress from Alfalfa

  • Houcong JinAffiliated withInstitute of Animal Sciences, Chinese Academy of Agricultural SciencesCollege of Agronomy, Inner Mongolia Agricultural University
  • , Yan SunAffiliated withCollege of Animal Science and Technology, China Agriculture University
  • , Qingchuan YangAffiliated withInstitute of Animal Sciences, Chinese Academy of Agricultural Sciences Email author 
  • , Yuehui ChaoAffiliated withInstitute of Animal Sciences, Chinese Academy of Agricultural Sciences
  • , Junmei KangAffiliated withInstitute of Animal Sciences, Chinese Academy of Agricultural Sciences
  • , Hong JinAffiliated withCollege of Agronomy, Inner Mongolia Agricultural University
  • , Yan LiAffiliated withInstitute of Animal Sciences, Chinese Academy of Agricultural Sciences
  • , Gruber MargaretAffiliated withSaskatoon Research Centre, Agriculture and Agri-Food

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Abstract

An alfalfa cDNA library induced by salt stress was constructed by suppression subtraction hybridization (SSH) technology. Total RNA from 10-day-old seedlings was used as a “driver,” and total RNA from seedlings induced by salt was used as a “tester”. One hundred and nineteen clones identified as positive clones by reverse Northern dot-blotting resulted in 82 uni-ESTs comprised of 16 contigs and 66 singletons. Blast analysis of deduced protein sequences revealed that 51 ESTs had identity similar to proteins with known function, while 24 could not be annotated at all. Most of the annotated sequences were homologous to genes involved in abiotic or biotic stress in plants. Among these proteins, beta-amylase, fructose-1,6-bisphosphate, aldolase, and sucrose synthase are related to osmolyte synthesis; a CCCH-type zinc finger protein, DNA binding protein, His–Asp phosphotransfer protein, and the RelA/SpoT protein partake in transcription regulation and signal transduction; and ribulose-l,5-bisphosphate carboxylase/oxygenase, chlorophyll a/b binding proteins, and an early light-inducible proteins are related to photosynthesis. In addition, several ESTs, similar to genes from other plant species, closely involved in salt stress were isolated from alfalfa, such as an aquaporin protein, a late embryogenesis-abundant protein, and glutathione peroxidase.

Keywords

Suppression subtractive hybridization (SSH) Alfalfa (Medicago sativa L.) SMART technology Reverse Northern dot-blotting Salt stress Real-time PCR