Molecular Cloning and Sequence Analysis of a Tonoplast Aquaporin Gene TIP1 Associated with Tapping P

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[Objective]To address the role of aquaporins(AQPs)in the occurrence of Tapping Panel Dryness(TPD),a phenomenon with tapping incision blocked partly or entirely during latex exploiting in the rubber tree(Hevea brasiliensis)that causes great losses on rubber production,a tonoplast aquaporin gene associated with TPD occurence was cloned and analyzed.[Method]Based on an EST down-regulated in TPD-affected rubber trees,a 774 bp cDNA designated HbTIP1 was isolated from the bark tissue of Hevea brasiliensis with a combination of in silico cloning and RT-PCR.And the gene structure and sequence characteristics were analyzed using bioinformatics tools.[Result]The cDNA includes a 759 bp ORF,8 bp 5’UTR and 7 bp 3’UTR.Sequence analysis indicated that HbTIP1putatively encodes 252 amino acids with a theoretical molecular weight(Mw)of 25.88 kDa and isolectric point(pI)of 4.96.Bioinformatics analysis suggested that the deduced protein is predicted to have six transmembrane helices located to the vacuolar membrane and to harbor one conserved MIP domain that can be grouped into the tonoplast intrinsic proteins(TIPs)of aquaporin family.Homology search revealed that the protein shares a similarity of more than 90%with the homologues in Theobroma cacao,Prunus persica,Citrus sinensis and Ricinus communis,supporting a highly conserved evolution.[Conclusion]This study provided basis for further revealing the regulatory role of AQPs in the TPD occurrence. [Objective] To address the role of aquaporins (AQPs) in the occurrence of Tapping Panel Dryness (TPD), a phenomenon with tapping incision blocked partly or entirely during latex exploiting in the rubber tree (Hevea brasiliensis) that causes great losses on rubber production , a tonoplast aquaporin gene associated with TPD occurence was cloned and analyzed. [Method] Based on an EST down-regulated in TPD-affected rubber trees, a 774 bp cDNA designated HbTIP1 was isolated from the bark tissue of Hevea brasiliensis with a combination of in silico cloning and RT-PCR. And the gene structure and sequence characteristics were analyzed using bioinformatics tools. [Result] The cDNA includes a 759 bp ORF, 8 bp 5 ’UTR and 7 bp 3’ UTR. Sequence analysis indicated that HbTIP1 inputatively encodes 252 amino acids with a theoretical molecular weight (Mw) of 25.88 kDa and isolectric point (pI) of 4.96. Bioinformatics analysis suggested that the deduced protein is predicted to have six transmembrane helices located to the vacuole r membrane and to harbor one conserved MIP domain that can be grouped into the tonoplast intrinsic proteins (TIPs) of aquaporin family. Homology searched revealed that the protein shares a similarity of more than 90% with the homologues in Theobroma cacao, Prunus persica, Citrus sinensis and Ricinus communis, supporting a highly conserved evolution. [Conclusion] This study provides basis for further revealing the regulatory role of AQPs in the TPD occurrence.
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