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Biochemical characterization of caffeoyl coenzyme A 3-O-methyltransferase from wheat

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Abstract

Main conclusion

TaCCoAOMT1 is located in wheat chromosome 7A and highly expressed in stem and root. It is important for lignin biosynthesis, and associated with stem maturity but not lodging resistance.

Caffeoyl coenzyme A 3-O-methyltransferases (CCoAOMTs) are one important class of enzymes to carry out the transfer of the methyl group from S-adenosylmethionine to the hydroxyl group, and play important roles in lignin and flavonoids biosynthesis. In the present study, sequences for CCoAOMT from the wheat genome were analyzed. One wheat CCoAOMT that belonged to bona fide subclade involved in lignin biosynthesis, namely TaCCoAOMT1, was obtained by the prokaryotic expression in E. coli. The three-dimensional structure prediction showed a highly similar structure of TaCCoAOMT1 with MsCCoAOMT. Recombinant TaCCoAOMT1 protein could only use caffeoyl CoA and 5-hydroxyferuloyl CoA as effective substrates and caffeoyl CoA as the best substrate. TaCCoAOMT1 had a narrow optimal pH and thermal stability. The TaCCoAOMT1 gene was highly expressed in wheat stem and root tissues, paralleled CCoAOMT enzyme activity. TaCCoAOMT1 mRNA abundance and enzyme activity increased linearly with stem maturity, but showed little difference between wheat lodging-resistant (H4546) and lodging-sensitive (C6001) cultivars in elongation, heading and milky stages. These data suggest that TaCCoAOMT1 is an important CCoAOMT for lignin biosynthesis that is critical for stem development, but not directly associated with lodging-resistant trait in wheat.

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Abbreviations

CCoAOMT:

Caffeoyl coenzyme A 3-O-methyltransferase

COMT:

Caffeic acid 3-O-methyltransferase

EST:

Expressed sequence tag

NCBI:

National Center for Biotechnology Information

OMT:

O-Methyltransferase

ORF:

Open reading frame

PCR:

Polymerase chain reaction

SAM:

S-Adenosylmethionine

TaGI:

Triticum aestivum gene index database

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Acknowledgments

This work was supported by grants from the National Natural Science Foundation of China (No. 31370336 and No. 31170277), the National Key Basic Research Program of China (973 program), the Natural Science Foundation of Beijing (No. 5122024), and the Innovation Project of Chinese Academy of Sciences.

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Correspondence to Qing-Hu Ma.

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Ma, QH., Luo, HR. Biochemical characterization of caffeoyl coenzyme A 3-O-methyltransferase from wheat. Planta 242, 113–122 (2015). https://doi.org/10.1007/s00425-015-2295-3

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