Analysis of Physiological Mechanism of Drought Tolerance in SbPIP1 Transgenic Tobacco

doi:10.7668/hbnxb.2015.06.003

Abstract

Abstract: In order to study the function of aquaporin genes from Salicornia bigeloii ,a new plasma membrane aquaporin gene SbPIP1 was cloned from S.bigeloii based on the rapid amplification of cDNA ends (RACE) technique and the amino acid sequence encoded by it was analyzed using bioinformatics software.The SbPIP1 gene driven by constitutive promoter CaMV 35S was transformed into tobacco NC89 with Agrobaeterium EHA105 containing the plant expression vector pcAMBIA2301- SbPIP1.The transgenic tobacco plants were screened by Kan resistance,PCR and semi-quantitative RT-PCR.Drought tolerance of transgenic tobacco plants was detected,and relative water content (RWC),malondialdehyde (MDA),Ion leakage (IL) and proline content of leaves of transgenic tobacco treated with different concentrations of PEG6000 were measured.The results showed that the full-length cDNA of SbPIP1 gene (GenBank Accession No.DQ451602) consists of 1 268 bp and contains a 858 bp open reading frame (ORF) encoding 285 amino acid.Bioinformatics analysis showed that SbPIP1 protein contained a typical structure with six membrane-spanning domains,two highly conserved Asn-Pro-Ala (NPA) motifs and two topical signal consensus sequence of plasma membrane.10 lines of positive transgenic tobacco were finally identified by Kan resistance,PCR and semi-quantitative RT-PCR.Transgenic plants which overexpressed SbPIP1 displayed improved tolerance against drought stress when compared to WT tobacco plants.Burst time of protoplasts from transgenic tobacco plants in hypotonic solution was significantly shorter than the control tobacco.Under PEG6000 treatments with different concentrations,the RWC in transgenic and control plants decreased gradually with the increase of the treatment concentration,however,at the same treatment concentration,the RWC of transgenic tobacco leaves were higher than that of control plants.With the increase of the treatment concentration of PEG6000,MDA and IL of the transgenic and the control plants increased gradually,but those of the transgenic plants increased more slowly than those of control plants.With the increase of the PEG concentration,the proline content of transgenic and control plants also gradually increased,but proline content of the transgenic plants were higher than those of control plants.

Key words: SbPIP1 gene, Tobacco, Transgene, Drought tolerance, Physiological index

CLC Number:

SUN Xiao-bo, JIA Xin-ping, LIANG Li-jian, DENG Yan-ming. Analysis of Physiological Mechanism of Drought Tolerance in SbPIP1 Transgenic Tobacco[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2015, 30(6): 17-26. doi: 10.7668/hbnxb.2015.06.003.

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