Cloning And Expression Analysis Of CsCaMs、CsCDPKs Family Genes In Tea Plant(Camellia Sinensis)

Cold stress is one of the most severe abiotic stresses greatly impacting on plant production. Tea plant（Camellia sinensis） is originated in southwestern region of China, which is thermophilic and sensitive to low temperature. Cold stress affects the production of tea, and also limits the expanding of tea planting areas to high latitude. Thus improving cold hardiness and cold resistance by variety breeding are the hot issue in tea plant study. Under ar tif ic ial low temperatur e stress, the changes of several cold r es istanc e related indexes w ere measur ed in in plants w ith or w ithout exogenous Ca Cl₂, Ca M inhibitor W7 [N-（6- Aminohexyl）- 5-chlor o-1- naphthalenes ulf onamide, HCl], and calcium s ignal channel inhibitor La Cl₃. Furthermore, cloning and expression analys is of Cs Ca Ms and Cs CDPKs were performed. This will provide helpful evidences for exploring the relationship between calcium signal channel and cold hardiness in tea plant.1. The changes of s everal cold r es istance r elated index es were meas ured under low temper ature str ess with exogenous Ca Cl₂, Ca M inhibitor W7 and calc ium s ignal channel inhibitor La Cl₃ treatments res pectively. W7 and La Cl₃ impr oved permeability of Plasma membrane（relative electrolyte leakage）, enhanced contents of malondialdehyde（ MDA）, superoxide anion radic al（O2·-）, reduced the activities of superoxide dismutase（S OD） compar ed w ith control（without treatments）. However, the Ca Cl₂ treatment had oppos ite eff ects. These indicat ed that calcium signaling system plays an important role in r esponse to low temper atur e stress in tea plant. Both the concentration of Ca²⁺ and activity of Ca M might affect the calcium signal transduction.2. Two full- length c DNA sequences of Cs Ca Ms were obtained using RACE-PCR and RT-PCR on the basis of cold acclimation transcriptome sequencing. They were named as Cs Ca M 1 and Cs Ca M2.The tissue spec if ic express ion of them has no signif icant differences; their expressions were induced under low temperature stress, were suppressed after the treatments of W7, La Cl₃ under low temperature stress, and increased sharply after applying the exogenous Ca²⁺. Above results showed that Cs Ca Ms and calmodulin play a role in maintaining the cold hardiness. 3. Twelve full-length c DNA sequences of Cs CDPKs, grouped into four subfamilies, were obtained by RACE-PCR and RT-PCR on the basis of cold acclimation transcriptome sequencing. Calc ium-dependent protein kinases were usually composed by Ca M and protein kinases, in which the structures of EF-hand and protein kinase domain were conservative. The different expression levels of Cs CDPK genes under low temperature demonstrated the variation of Cs CDPK genes in expression and regulation patterns. Cs CDPK genes also had different responses to the treatments of Ca Cl₂, W7 and La Cl₃ under low temperature stress. It is implied that calcium-dependent protein kinase and calmodulin might not only be regulated by single factor in calcium signal pathway, moreover, various physiological processes could be regulated by calcium in tea plant.