• C C Chinnappa

      Articles written in Journal of Biosciences

    • Molecular characterization of a cDNA encoding caffeoyl-coenzyme A 3-O-methyltransferase ofStellaria longipes

      Xing-Hai Zhang C C Chinnappa

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      A cDNA clone encodingS-adenosyl-L-methionine:trans-caffeoyl-CoA 3-O-methyl-transferase (EC; CCoAOMT) fromStellana longipes Goldie (long-stalked chick-weed) was isolated and studied. Structural analysis of both the nucleotide sequence and the predicted amino acid sequence suggests that our cloned sequence encoded a CCoAOMT enzyme ofStellaria longipes, which shared overall structural similarity with other plant CCoAOMTs but exhibited certain distinct characteristics. Southern blot hybridization and cloning analyses indicating a small CCoAOMT gene family in theStellana longipes genome and the absence of introns in the coding region of the cDNA-corresponding gene. Sequence variations in the coding region were found among three genotypes from geographically isolated populations. Higher levels of CCoAOMT mRNA were detected in stems and leaves than in roots. The cDNA-encoded protein expressed inEschendia coli was shown to utilize caffeoyl-CoA, but not caffeic acid or 5-hydroxy ferulic acid, as its substrate.

    • Cytosine methylation levels in the genome ofStellaria longipes

      Q Cai C C Chinnappa

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      Environment-induced alteration of DNA methylation levels was investigated inStellaria longipes (Caryophyllaceae). Total cytosine methylation levels were measured using HPLC in 6 genets representing two ecotypes (alpine and prairie) grown in short day photoperiod and cold temperature (SDC) and long day photoperiod and warm temperature (LDW) conditions. The levels of methylated cytosine were 16.54-22.20% among the three genets from the alpine and 12.62–24.70% in the three prairie genets when they were grown in SDC conditions. After the plants were moved to the LDW conditions, all of the three genets from the alpine showed decreasing levels of DNA methylation up to 6 days of growing in LDW. When the plants continued to grow in LDW for 10 days the average methylation level in the prairie genotypes showed no significant change. Cytosine methylation level was also detected inHpall andSau3AI restriction sites using the coupled restriction enzyme digestion and random amplification (CRED-RA) procedure, in which 15 random primers were used. Fifty per cent of the amplified bands with either or both of these two restriction sites were identified as being methylated in an alpine genotype (1C) and approximately 66% were found to be methylated in a prairie genotype (7C). It was observed that the change in growing conditions from SDC to LDW induced a decrease of methylation levels inHpall sites.

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