The Calcium Metabolism-Related Gene Cloning And Expression Analysis Of Lonicera Confusa L

The weathering of carbonate rock in the Karst area of southwest China results in the abundance of calcium in this Karst soil ecosystem, however Lonicera japonica L adapts well to this kind of soil condition. Lonicera japonica L, a sort of liana, has strong root system and reproductive capacity, which plays particularly important roles in preventing from soil erosion and restoring the local ecosystem. A great deal of scientific research proved that Lonicera confusa L is able to grow well in the Kast area, which is closely related to the function of calcium excretion. According to the Lonicera confusa L's analysis of calcium-related gene sequence and expression through the soil environment's simulation with different concentrations of calcium, hereby we obtained the following results:(1) For both cloned Lonicera confusa & Lonicera japonica, the Calmodulin cDNA sequence's full-length is 450bp, encoding 150 amino acids. Using the Blast analysis, the homology is over 90% comparing to other reported species. The genetic homology rate is 98% between Lonicera and Camellia oleifera; is 95% between Lonicera and Solanum tuberosum; is 96% between Lonicera and Catharanthus roseus, is 93% between Lonicera, Zea mays, wheat (Triticum aestirum), rice (Oryza sativa) and other monocotyledons. The comparative analysis of proteins proves the homology is even higher than other species of calmodulin, and only found few individual amino acid sites mutation, which illustrates the high conservation of calmodulin. After measuring and analyzing the sequence of Lonicera confusa's calmodulin DNA, the total length of 2053bp was worked out, also found there was only one copy for the calmodulin DNA sequence.(2) With fluorescence quantitative PCR testing on calmodulin by different calcium concentration, the experiment showed that with the increased calcium concentration, gene expression was also increased for both Lonicera confuse & Lonicera japonica, but Lonicera japonica increased more significantly than Lonicera confusa.(3) The cDNA sequence full length of the cloned Lonicera confusa calmodulin protein kinase (CDPK) is 960bp, encoding 263 amino acids, with 85% homology to other reported species'CDPKs.(4) The cDNA sequence full length of the cloned Lonicera confusa transferring calcium ACA1 is 3057bp, encoding 1018 amino acids, with 80% homology to Arabidopsis' ACAl.