Cloning and Expression of Transcription Factor SpBBX1 in Solanum pennellii and Preliminary Analysis of Its Stress Tolerance
ZHAO Si-Fang1,2, ZHOU Tao1,2, HU Jia-Hui1,2, LAN Hai-Yan1,*, YU Qing-Hui2,*
1. Xinjiang Key Laboratory of Biological Resources and Genetic Engineering/College of Life Science and Technology, Xinjiang University, Urumqi 830046, China; 2. Institute of Horticultural Crops, Xinjiang Academy of Agriculture Sciences, Urumqi 830091, China
Abstract：B-box (BBX) transcription factor plays an important role in the photomorphogenesis of seedlings, regulation of flowering photoperiod, avoidance of light and other growth and developmental processes, as well as in response to the biotic and abiotic stresses. In this study, a BBX protein was obtained from Solanum pennellii and named as SpBBX1 (Gene ID: Sopen02g034260). In order to study its function, the physicochemical properties, protein structure, phylogenetic tree, subcellular localization and prokaryotic expression were analyzed. The results showed that SpBBX1 contained a complete open reading frame of 1 173 bp, encoding 390 amino acids, and was located in the nucleus; phylogenetic tree analysis showed its amino acid sequence was closely related to cultivated tomato protein SlCONSTANS1 (Gene ID: Solyc02g089540) and potato protein StB-box1 (Gene ID: PGSC0003DMP400017796); SDS-PAGE and Western blot results showed that the molecular weight of recombinant SpBBX1 protein was about 48 kD; qRT-PCR analysis found that SpBBX1 was expressed in stem, leaf, and flower, with the highest expression in leaf. The prokaryotic recombinant strain Escherichia coli BL21::pET-30a-SpBBX1 showed significantly lower growth than the control strain in response to NaCl or polyethylene glycol (PEG) stress (P<0.05). These results suggested that SpBBX1 gene may have a negative regulatory effect in response to stress, the data could provide a theoretical basis for further research on plant BBX function.
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