Chinese Bulletin of Botany ›› 2017, Vol. 52 ›› Issue (5): 568-578.DOI: 10.11983/CBB16225

• EXPERIMENTAL COMMUNICATIONS • Previous Articles     Next Articles

Expression Analysis of β-1,3-Glucanase Gene from Winter Brassica rapa Under Low Temperature Stress

Li Ma1, Wancang Sun1,*(), Jinhai Yuan1, Zigang Liu1, Junyan Wu1, Yan Fang1, Yaozhao Xu2, Yuanyuan Pu1, Jing Bai1, Xiaoyun Dong1, Huili He1   

  1. 1Gansu Key Laboratory of Crop Improvement and Germplasm Enhancement, Rapeseed Engineering Research Center of Gansu Province, College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China

    2College of Agronomy and Biotechnology, Hexi University, Zhangye 734000, China
  • Received:2016-11-23 Accepted:2017-03-24 Online:2017-09-01 Published:2017-07-10
  • Contact: Wancang Sun

Abstract: This study examined the function of β-1,3-glucanase in Brassica rapa with low temperature stress, whose encoding protein was identified by protein mass spectrometry analysis. The cDNA sequence of β-1,3-glucanase was cloned by RT-PCR of winter rapeseed from Longyou6 and Tianyou4 cultivars. Semi-quantitative PCR and real-time fluorescence quantitative PCR were used to study the expression pattern of β-1,3-glucanase in response to low temperature stress. The open reading frame of the β-1,3-glucanase gene was obtained at a length of 1 032 bp, and encoded 343 amino acid. The molecular weight was 38.102 kDa and the isoelectric point 6.63, with 93.94% amino sequence similarity to B. rapa subsp. chinensis and B. napus. The protein encoded by this gene is a hydrophilic protein with one signal peptide and two transmembrane domains. The prediction of the secondary structures indicated that β-1,3-glucanase is a protein with more α-helices. The predicted β-1,3-glucanase contains a conserved amino acid sequence corresponding to the plant Glycol-Hydro 17 superfamily. RT-PCR and semi-quantitative RT-PCR results showed that β-1,3-glucanase was upregulated in response to 4°C, and the gene was upregulated to peak in expression at -4°C; However, the expression was inhibited at lower temperature (-8°C). The β-1,3-glucanase gene cloned from the winter B. rapa cv. ‘Longyou6’ might play a role in cold tolerance.

Key words: β-1, 3-glucanase gene, low temperature, molecular cloning, expression analysis