Class I patatin genes from potato (Solanum tuberosum L.) cultivars: molecular cloning, sequence comparison, prediction of diverse cis-regulatory motifs, and assessment of the promoter activities under field and in vitro conditions
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Patatin, the most abundant glycoprotein (∼40 kDa) in potato (Solanum tuberosum L.) tuber is encoded by a large multigene family comprising of two classes. Class I patatin gene promoters are attractive molecular tools for studying multiple facets of transcriptional regulation. Only a few members of this class are relatively well understood at the molecular level. Here, we report the isolation of two new partial class I patatin genes with corresponding 5′-flanking regions, designated as StPN02 and StPK01, from two commercially important Indian potato cultivars, ‘Kufri Jyoti’ and ‘Kufri Chipsona-1’, respectively. Their 5′-flanking regions were found to vary significantly because of insertions/deletions in addition to point mutations. A major 487-bp insertion in the 5′-flanking region of StPN02 was a distinctive from other family members. Many of the diverse cis-regulatory sequence motifs predicted in this study have not been previously documented. The 1,215 and 2,225 bp 5′-flanking regions of StPN03 (derived from StPN02) and StPK01, respectively, were used in generating patatin promoter–GUS gene fusions for assessment of promoter activities in potato. Under field conditions, both StPK01 and StPN03 promoters showed tuber-specificity. The former was significantly stronger than the latter as examined by histochemical and fluorometric β-glucuronidase assays. Under in vitro conditions, these promoters were highly inducible by sucrose in stems and leaves. Apart from the applied aspects of this research, this study may help in unraveling the molecular mechanisms involved in the regulation of patatin gene expression.
KeywordsClass I patatin genes Putative cis-regulatory motifs Tuber-specificity Sucrose-inducible expression Indian potato cultivars
The authors are thankful to the University Grants Commission (UGC), Government of India, for providing “UGC-Meritorious Fellowship” to Raghavendra Aminedi.
- Genova AD, Goverse A, Massa AN, Beletsky AV, Mardanov AV, Kuznetsov BB, Kloosterman B, Whitty BR, Sagredo B, Hekkert BL, Vaillancourt B, Xie B, Zhang B, Liang C, Buell CR, Bachem CWB, Marshall DF, Martinez D, Milbourne D, Martin DMA, Bolser DM, DellaPenna D, Mu D, Datema E, Guzman F, Massa GA, Torres GA, Cruz GD, Giuliano G, Bishop GJ, Bryan GJ, Li G, Orjeda G, Perrotta G, Zhang G, Kuang H, Lin H, Eck H, Nagy I, Boer J, Gawor J, He J, Jiang J, Jacobs JME, Wang J, Xu J, Nielsen KL, O’Brien K, Diambra L, Barreiro LE, Bonierbale M, Destefanis M, Herrera MR, Fiers M, Geoffroy M, Ghislain M, Iovene M, Pietrella M, Sønderkær M, Torres M, Mejı’a N, Ravin NV, Ponce O, Facella P, Ni P, Szczesny P, Hu Q, Ham RCHJ, Veilleux RE, Gromadka R, Visser RGF, Lozano R, Li R, Cheng S, Feingold SE, Huang S, Thomson SJ, Chakrabarti SK, Sharma SK, Lundback S, Pan S, Yang S, Borm T, Kolganova TV, Patil VU, Xiong X, Xu X, Du Y, He Y, Li Y, Yang Y, Zhang Y, Zhang Z, Waugh R, Qu D, Skryabin KG, Zagorski W (2011) Genome sequence and analysis of the tuber crop potato. Nature 475:189–195PubMedCrossRefGoogle Scholar
- Kumari V, Bansal A, Aminedi R, Taneja D, Das N (2012) Simplified extraction of good quality genomic DNA from a variety of plant materials. Afr J Biotechnol 11:6420–6427Google Scholar
- Nakashima K, Fujita Y, Katsura K, Maruyama K, Narusaka Y, Seki M, Shinozaki K, Yamaguchi-Shinozaki K (2006) Transcriptional regulation of ABI3- and ABA-responsive genes including RD29B and RD29A in seeds, germinating embryos, and seedlings of Arabidopsis. Plant Mol Biol 60:51–68PubMedCrossRefGoogle Scholar
- Park WD (1993) Regulation and evolution of the patatin genes of potato. In: Verma DPS (ed) Control of plant gene expression. CRC Press, Boca Raton, pp 393–405Google Scholar
- Racusen D (1985) Patatin and esterase in Desiree potato tuber. J Food Ind 9:361–367Google Scholar
- Sambrook J, Fritsch EF, Maniatis T (1989) Molecular cloning: a laboratory manual, 2nd edn. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, 2626 ppGoogle Scholar