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Molecular cloning, characterization, and expression analysis of lignin biosynthesis genes from kenaf (Hibiscus cannabinus L.)

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Abstract

Kenaf (Hibiscus cannabinus L.) is an annual herbaceous crop and the third largest fiber crop of economic importance, after cotton and jute. In view of recent global environmental challenges, multipurpose uses of kenaf have been developed, such as animal feed, bio-plastics, bio-fuels, and phytoremediation. We cloned genes related to monolignol biosynthesis using a reverse transcriptase PCR (RT-PCR) strategy. ‘Auxu’ cultivars in the seedling phase were used as the experimental source material. The 4-coumarate:CoAligase (4CL), cinnamoyl-CoA reductase (CCR), ferulate 5-hydroxylase (F5H), caffeic acid O-methyltransferase (COMT), cinnamyl alcohol dehydrogenase (CAD), encoding hydroxycinnamoyl transferase (HCT), ρ-coumarate 3-hydroxylase (C3H) and caffeoyl-CoA-O-methyltransferase (CCoAOMT), cDNAs had open reading frames of 1623, 1017, 1179, 921, 1107, 1323, 1527 and 744 bp, encoding polypeptides of 540, 338, 392, 306, 368, 441, 508 and 247 amino acid residues, respectively. We submitted the five gene sequences (4CL, CCR, F5H, COMT, and CAD) to the GenBank database. A phylogenetic tree showed that lignin biosynthesis genes had the closest relationships with Gossypium and Theobroma cacao. Expression levels of the genes 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD in stems from plants harvested at four different growth stages were evaluated. Expression levels of 4CL, HCT, C3H, CCoAOMT, CCR, F5H, COMT, and CAD were gradually increased during the vegetative growth of kenaf stem. Similarly with the gene expression, lignin amounts were also increased during the stem growth. Thus, these six genes may be directly involved in lignin biosynthesis. These results will be useful basic data for molecular breeding to enhance bio-mass production.

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Acknowledgments

This work was supported by a grant from the Korea Atomic Energy Research Institute (KAERI), Republic of Korea.

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    Authors and Affiliations

    1. Advanced Radiation Technology Institute, Korea Atomic Energy Research Institute, Jeongup, Jeonbuk, 580-185, Korea

      Jaihyunk Ryu, Soon-Jae Kwon, Sang Yeop Sung, Wook-Jin Kim, Dong Sub Kim, Joon-Woo Ahn, Jin-Beak Kim, Sang Hoon Kim, Bo-Keun Ha & Si-Yong Kang

    2. Division of Plant Biotechnology, College of Agriculture and Life Science, Chonnam National University, Gwangju, 500-757, Korea

      Bo-Keun Ha

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    1. Jaihyunk Ryu
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    Correspondence to Bo-Keun Ha or Si-Yong Kang.

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    Jaihyunk Ryu and Soon-Jae Kwon contributed equally to this work.

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    13258_2015_341_MOESM1_ESM.bmp

    Supplementary material 1 (BMP 3773 kb). Supplement Fig. 1. Multiple alignment of the deduced amino acid sequences of the kenaf lignin orthologue. a 4CL, b CCR, c F5H, d COMT, e CAD. GenBank accession numbers are represented as follows; 1 ADK24217, 2 AGJ84134, 3 ADK24219, 4 AGJ84130, 5 AGR 85825, 6 AGJ84132, 7 AGJ84131, 8 AGR85824, 9 ADK24218, 10 ABG81259

    Supplementary material 2 (DOCX 18 kb)

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    Ryu, J., Kwon, SJ., Sung, S.Y. et al. Molecular cloning, characterization, and expression analysis of lignin biosynthesis genes from kenaf (Hibiscus cannabinus L.). Genes Genom 38, 59–67 (2016). https://doi.org/10.1007/s13258-015-0341-y

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