Expression patterns and salt tolerance analysis of BpPAT1 gene in Betula platyphylla

  Objective  GRAS family is a plant-specific transcription factor family, characterized by a highly conserved carboxyl terminus domain. Previous studies have shown that GRAS transcription factor is one of the key transcriptional regulators in plant stress response. The purpose of this study is to analyze the salt tolerance of GRAS transcription factor gene BpPAT1 gene in Betula platyphylla, so as to lay a foundation for elucidating the molecular regulation mechanism of GRAS transcription factor in response to salt stress. Our work enriched the research on the molecular mechanism of the GRAS transcription factors of woody plant in response to stress.

  Method  In this study, one GRAS transcription factor gene was screened from the transcriptome data of B. platyphylla under salt stress and named as BpPAT1. Multiple sequence alignment and phylogenetic tree were used to analyze the genetic relationship between BpPAT1 and other organism’s GRAS family genes. Real-time fluorescence quantitative PCR (qRT-PCR) method was used to analyze the expression pattern of BpPAT1 in root, stem and leaf tissues of B. platyphylla under salt stress and normal condition, to identify whether it responded to salt stress or not. In order to further analyze the stress tolerance function of BpPAT1, plant overexpression vector (pROKII-BpPAT1) and inhibitory expression vector (pFGC5941-BpPAT1) were constructed. Transient overexpression and inhibitory expression of BpPAT1 gene and control B. platyphylla plants were obtained by Agrobacterium tumefaciens-mediated transient genetic transformation system. The physiological indexes related to salt tolerance were measured to identify whether the BpPAT1 was associated with salt tolerance in transient expression of BpPAT1 and control plants under salt stress.

  Result  The results of multiple sequence alignment and phylogenetic tree analysis showed that BpPAT1 protein had the sequence characteristics of GRAS family and was closely related to AtPAT1 protein in A. thaliana. The result level of qRT-PCR showed that the expression of BpPAT increased significantly in B. platyphylla plants after 6 hours of salt stress, indicating that BpPAT1 could respond to salt stress signal. The measurement results of the physiological indexes of stress resistance showed that the overexpression of BpPAT1 in B. platyphylla could significantly increase the activity of peroxidase (POD) and superoxide dismutase (SOD), increased the content of proline, and decreased electrolyte leakage and malondialdehyde (MDA) content.

  Conclusion  The BpPAT1 gene can respond to salt stress, overexpression of BpPAT1 significantly enhances POD, SOD enzyme activities and proline content, decreases electrolyte leakage and MDA content under salt stress, thus improves ROS scavenging ability and salt tolerance of B. platyphylla.