title: Structure and expression profiling of a novel calcium-dependent protein kinase gene, CDPK3a, in leaves, stems, grapes, and cell cultures of wild-growing grapevine Vitis amurensis Rupr.
abstract: KEY MESSAGE                 :                                     VaCDPK3a                   is actively expressed in leaves, stems, inflorescences, and berries of                   Vitis amurensis                   and may act as a positive growth regulator, but is not involved in the regulation of resveratrol biosynthesis.                                                                  Calcium-dependent protein kinases (CDPKs) are known to play important roles in plant development and defense against biotic and abiotic stresses. It has previously been shown that CDPK3a is the predominant CDPK transcript in cell cultures of wild-growing grapevine Vitis amurensis Rupr., which is known to possess high resistance against environmental stresses and to produce resveratrol, a polyphenol with valuable pharmacological effects. In this study, we aimed to define the full cDNA sequence of VaCDPK3a and analyze its organ-specific expression, responses to plant hormones, temperature stress and exogenous NaCl, and the effects of VaCDPK3a overexpression on biomass accumulation and resveratrol content in V. amurensis calli. VaCDPK3a was actively expressed in all analyzed V. amurensis organs and tissues and was not transcriptionally regulated by salt and temperature stresses. The highest VaCDPK3a expression was detected in young leaves and the lowest in stems. A reduction in the VaCDPK3a expression correlated with a lower rate of biomass accumulation and higher resveratrol content in calli of V. amurensis under different growth conditions. Overexpression of the VaCDPK3a gene in the V. amurensis calli significantly increased cell growth for a short period of time but did not have an effect on resveratrol production. Further subculturing of the transformed calli resulted in cell death and a decrease in expression of the endogenous VaCDPK3a. The data suggest that while VaCDPK3a acts as a positive regulator of V. amurensis cell growth, it is not involved in the signaling pathway regulating resveratrol biosynthesis and resistance to salt and temperature stresses.
authors.affiliation: Laboratory of Biotechnology, Institute of Biology and Soil Science, Far Eastern Branch of the Russian Academy of Sciences, Vladivostok 690022, Russia. kiselev@biosoil.ru
authors.forename: K V
authors.initials: KV
authors.lastname: Kiselev
authors.forename: A S
authors.initials: AS
authors.lastname: Dubrovina
authors.forename: O A
authors.initials: OA
authors.lastname: Shumakova
authors.forename: Y A
authors.initials: YA
authors.lastname: Karetin
authors.forename: A Y
authors.initials: AY
authors.lastname: Manyakhin
mesh_terms: D001483:Base Sequence; D018533:Biomass; D018929:Cell Culture Techniques; D018076:DNA, Complementary / Q000235:genetics; D005638:Fruit / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D015870:Gene Expression; D018507:Gene Expression Regulation, Developmental / Q000235:genetics; D018506:Gene Expression Regulation, Plant / Q000235:genetics*; D056626:Inflorescence / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D008969:Molecular Sequence Data; D010802:Phylogeny; D018515:Plant Leaves / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D010940:Plant Proteins / Q000235:genetics / Q000378:metabolism; D018547:Plant Stems / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D030821:Plants, Genetically Modified; D011494:Protein Kinases / Q000235:genetics* / Q000378:metabolism; D018749:RNA, Plant / Q000235:genetics; D000077185:Resveratrol; D012492:Salts; D012639:Seeds / Q000201:enzymology / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D017422:Sequence Analysis, DNA; D013267:Stilbenes / Q000378:metabolism; D013312:Stress, Physiological; D013696:Temperature; D027843:Vitis / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development / Q000502:physiology; D017871:Calcium-Calmodulin-Dependent Protein Kinases
chemical_list: D018076:DNA, Complementary; D010940:Plant Proteins; D011494:Protein Kinases; D018749:RNA, Plant; D000077185:Resveratrol; D012492:Salts; D013267:Stilbenes; D017871:Calcium-Calmodulin-Dependent Protein Kinases