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Bulletin of Botanical Research ›› 2024, Vol. 44 ›› Issue (6): 914-925.doi: 10.7525/j.issn.1673-5102.2024.06.012

• Molecular biology • Previous Articles     Next Articles

Cloning and Drought Tolerance Functional Validation of PvAQP Gene in Psammochloa villosa (Poaceae)

Yuping LIU1,2,3, Mingjun YU1,2, Yu ZHANG1,2, Xu SU1,2,3(), Xiaoli LI1,2, Qian YANG1,2, Xueli LIU1,2   

  1. 1.School of Life Sciences,Qinghai Normal University,Xining 810008
    2.Key Laboratory of Biodiversity Formation Mechanism and Comprehensive Utilization of the Qinghai-Xizang Plateau in Qinghai Province,Qinghai Normal University,Xining 810008
    3.Academy of Plateau Science and Sustainability,Qinghai Normal University,Xining 810016
  • Received:2024-03-06 Online:2024-11-20 Published:2024-11-22
  • Contact: Xu SU E-mail:xusu8527972@126.com

Abstract:

In order to clarify the function and role of the AQP gene of Psammochloa villosa in response to drought tolerance, the AQP gene of P. villosa was cloned by using the three generations full transcriptome data and RT-PCR, and the bioinformatics and expression patterns of AQP gene under drought stress and drought tolerance in transgenic tobacco(Nicotiana tabacum) were analyzed, respectively. The results indicated that: (1)the AQP gene of P. villosa was successfully cloned, named PvAQP(GenBank number: ON792207), CDS length of PvAQP was 867 bp, encoding 288 amino acids and belonging to hydrophobic protein. (2)PvAQP had obvious transmembrane structure with a complete MIP domain, which was a transmembrane protein. (3)The secondary structure of PvAQP encoded protein included α-helix, irregular curls, extended chains and β-fold, while the tertiary structure was mainly composed of α-helix and irregular curls. (4)PvAQP gene had no signal peptide structure, no glycosylation sites, and contained 19 phosphorylation sites. (5)Subcellular localization analysis showed that PvAQP protein was localized in the nucleus. (6)Among different species, the homology with amino acid sequences of PvAQP gene was 95.34%. P. villosa had the closest genetic relationship with Stipa caucasica. (7)The expression of PvAQP gene was tissue-specific, with significant differences in the roots, stems, and leaves of P. villosa. (8)With the increase of simulated drought-stress time, the activities of superoxide dismutase(SOD), peroxidase(POD) and catalase(CAT), and the content of proline(PRO) first increased and then decreased in transgenic tobacco and wild type tobacco. Compared with wild type tobacco, the express levels in the transgenic tobacco were higher, while the content of malondialdehyde(MDA) showed an increasing trend, and the content of MDA in transgenic PvAQP gene-type was significantly lower than that of wild type. The results suggested that PvAQP gene might be involved in the physiological process of drought tolerance within P. villosa, and provided the theoretical evidence for revealing the molecular mechanism of PvAQP gene responding to drought tolerancein the future.

Key words: Psammochloa villosa, AQP gene, gene cloning, drought stress, functional validation

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