Isolation and characterization of CCoAOMT in interspecific hybrid of Acacia auriculiformis x Acacia mangium - A key gene in lignin biosynthesis

S. L. Pang, S. S. Ong, H. H. Lee, Zamri Zainal, K. I. Kandasamy, Chee Yen Choong, R Wickneswari V Ratnam

Research output: Contribution to journalArticle

8 Citations (Scopus)

Abstract

This study was directed at the understanding of the function of CCoAOMT isolated from Acacia auriculiformis x Acacia mangium. Full length cDNA of the Acacia hybrid CCoAOMT (AhCCoAOMT) was 1024-bp long, containing 750-bp coding regions, with one major open reading frame of 249 amino acids. On the other hand, full length genomic sequence of the CCoAOMT (AhgflCCoAOMT) was 2548 bp long, containing three introns and four exons with a 5' untranslated region (5'UTR) of 391 bp in length. The 5'UTR of the characterized CCoAOMT gene contains various regulatory elements. Southern analysis revealed that the Acacia hybrid has more than three copies of the CCoAOMT gene. Real-time PCR showed that this gene was expressed in root, inner bark, leaf, flower and seed pod of the Acacia hybrid. Downregulation of the homologous CCoAOMT gene in tobacco by antisense (AS) and intron-containing hairpin (IHP) constructs containing partial AhCCoAOMT led to reduction in lignin content. Expression of the CCoAOMT in AS line (pART-HAS78-03) and IHP line (pART-HIHP78-06) was reduced respectively by 37 and 75% compared to the control, resulting in a decrease in the estimated lignin content by 24 and 56%, respectively. AhCCoAOMT was found to have altered not only S and G units but also total lignin content, which is of economic value to the pulp industry. Subsequent polymorphism analysis of this gene across eight different genetic backgrounds each of A. mangium and A. auriculiformis revealed 47 single nucleotide polymorphisms (SNPs) in A. auriculiformis CCoAOMT and 30 SNPs in A. mangium CCoAOMT.

Original languageEnglish
Pages (from-to)7217-7238
Number of pages22
JournalGenetics and Molecular Research
Volume13
Issue number3
DOIs
Publication statusPublished - 5 Sep 2014

Fingerprint

caffeoyl-CoA O-methyltransferase
Acacia
Lignin
Genes
Introns
5' Untranslated Regions
Single Nucleotide Polymorphism

Keywords

  • Acacia hybrid
  • CCoAOMT
  • Downregulation
  • Lignin biosynthesis
  • Single nucleotide polymorphism

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

Cite this

Isolation and characterization of CCoAOMT in interspecific hybrid of Acacia auriculiformis x Acacia mangium - A key gene in lignin biosynthesis. / Pang, S. L.; Ong, S. S.; Lee, H. H.; Zainal, Zamri; Kandasamy, K. I.; Choong, Chee Yen; V Ratnam, R Wickneswari.

In: Genetics and Molecular Research, Vol. 13, No. 3, 05.09.2014, p. 7217-7238.

Research output: Contribution to journalArticle

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abstract = "This study was directed at the understanding of the function of CCoAOMT isolated from Acacia auriculiformis x Acacia mangium. Full length cDNA of the Acacia hybrid CCoAOMT (AhCCoAOMT) was 1024-bp long, containing 750-bp coding regions, with one major open reading frame of 249 amino acids. On the other hand, full length genomic sequence of the CCoAOMT (AhgflCCoAOMT) was 2548 bp long, containing three introns and four exons with a 5' untranslated region (5'UTR) of 391 bp in length. The 5'UTR of the characterized CCoAOMT gene contains various regulatory elements. Southern analysis revealed that the Acacia hybrid has more than three copies of the CCoAOMT gene. Real-time PCR showed that this gene was expressed in root, inner bark, leaf, flower and seed pod of the Acacia hybrid. Downregulation of the homologous CCoAOMT gene in tobacco by antisense (AS) and intron-containing hairpin (IHP) constructs containing partial AhCCoAOMT led to reduction in lignin content. Expression of the CCoAOMT in AS line (pART-HAS78-03) and IHP line (pART-HIHP78-06) was reduced respectively by 37 and 75{\%} compared to the control, resulting in a decrease in the estimated lignin content by 24 and 56{\%}, respectively. AhCCoAOMT was found to have altered not only S and G units but also total lignin content, which is of economic value to the pulp industry. Subsequent polymorphism analysis of this gene across eight different genetic backgrounds each of A. mangium and A. auriculiformis revealed 47 single nucleotide polymorphisms (SNPs) in A. auriculiformis CCoAOMT and 30 SNPs in A. mangium CCoAOMT.",
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