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Overexpression of hydroperoxide lyase gene in Nicotiana benthamiana using a viral vector system.
Plant Biotechnol J. 2010 Sep; 8(7):783-95.PB

Abstract

13-Lipoxygenase (13-LOX) and 13-hydroperoxide lyases (13-HPL) are the key enzymes for the production of the 'green note' compounds hexanal, (3Z)- and (2E)-hexenal in plant tissues. To produce high levels of 13-LOX and 13-HPL enzymatic activities for a biocatalytic process to generate C(6)-aldehydes on a large scale, soya bean 13-LOX (GmVLXC) and watermelon 13-HPL (ClHPL) genes were expressed in Nicotiana benthamiana using a viral vector system mediated by agroinfiltration. The N. benthamiana leaves produced high activity of watermelon HPL, but not GmVLXC 13-LOX. In addition, all leaves treated with bacterial suspension displayed a high activity of 9-LOX, indicating that the internal tobacco 9-LOX gene was highly induced through agroinfiltration because of wounding. GmVLXC and ClHPL transcripts could be detected in the corresponding transformed tobacco leaves by real-time RT-PCR analysis but the expression level of ClHPL was 24-fold higher than that of GmVLXC. Western blot analysis showed that LOX was present in all tobacco leaves which were treated with bacterial suspensions, but not in the untreated wild-type control. This result confirms that internal 9-LOX was highly induced by agroinfiltration. The highest levels of ClHPL activity under optimal infiltration conditions were 80 times the HPL activity of wild-type plants or plants transformed with control vector. A large amount of hexanal was formed when linoleic acid was incubated with extracts from N. benthamiana leaves over-expressing ClHPL in combination with GmVLXC-expressing yeast extracts. One gram of ClHPL-expressing N. benthamiana leaves (fresh weight) could produce 17 +/- 0.4 mg hexanal from 50 mg linoleic acid after 30 min.

Authors+Show Affiliations

Technische Universität München, Biomolecular Food Technology, Freising, Germany.No affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

20691022

Citation

Huang, Fong-Chin, et al. "Overexpression of Hydroperoxide Lyase Gene in Nicotiana Benthamiana Using a Viral Vector System." Plant Biotechnology Journal, vol. 8, no. 7, 2010, pp. 783-95.
Huang FC, Studart-Witkowski C, Schwab W. Overexpression of hydroperoxide lyase gene in Nicotiana benthamiana using a viral vector system. Plant Biotechnol J. 2010;8(7):783-95.
Huang, F. C., Studart-Witkowski, C., & Schwab, W. (2010). Overexpression of hydroperoxide lyase gene in Nicotiana benthamiana using a viral vector system. Plant Biotechnology Journal, 8(7), 783-95. https://doi.org/10.1111/j.1467-7652.2010.00508.x
Huang FC, Studart-Witkowski C, Schwab W. Overexpression of Hydroperoxide Lyase Gene in Nicotiana Benthamiana Using a Viral Vector System. Plant Biotechnol J. 2010;8(7):783-95. PubMed PMID: 20691022.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Overexpression of hydroperoxide lyase gene in Nicotiana benthamiana using a viral vector system. AU - Huang,Fong-Chin, AU - Studart-Witkowski,Claudia, AU - Schwab,Wilfried, PY - 2010/8/10/entrez PY - 2010/8/10/pubmed PY - 2010/10/29/medline SP - 783 EP - 95 JF - Plant biotechnology journal JO - Plant Biotechnol J VL - 8 IS - 7 N2 - 13-Lipoxygenase (13-LOX) and 13-hydroperoxide lyases (13-HPL) are the key enzymes for the production of the 'green note' compounds hexanal, (3Z)- and (2E)-hexenal in plant tissues. To produce high levels of 13-LOX and 13-HPL enzymatic activities for a biocatalytic process to generate C(6)-aldehydes on a large scale, soya bean 13-LOX (GmVLXC) and watermelon 13-HPL (ClHPL) genes were expressed in Nicotiana benthamiana using a viral vector system mediated by agroinfiltration. The N. benthamiana leaves produced high activity of watermelon HPL, but not GmVLXC 13-LOX. In addition, all leaves treated with bacterial suspension displayed a high activity of 9-LOX, indicating that the internal tobacco 9-LOX gene was highly induced through agroinfiltration because of wounding. GmVLXC and ClHPL transcripts could be detected in the corresponding transformed tobacco leaves by real-time RT-PCR analysis but the expression level of ClHPL was 24-fold higher than that of GmVLXC. Western blot analysis showed that LOX was present in all tobacco leaves which were treated with bacterial suspensions, but not in the untreated wild-type control. This result confirms that internal 9-LOX was highly induced by agroinfiltration. The highest levels of ClHPL activity under optimal infiltration conditions were 80 times the HPL activity of wild-type plants or plants transformed with control vector. A large amount of hexanal was formed when linoleic acid was incubated with extracts from N. benthamiana leaves over-expressing ClHPL in combination with GmVLXC-expressing yeast extracts. One gram of ClHPL-expressing N. benthamiana leaves (fresh weight) could produce 17 +/- 0.4 mg hexanal from 50 mg linoleic acid after 30 min. SN - 1467-7652 UR - https://www.unboundmedicine.com/medline/citation/20691022/Overexpression_of_hydroperoxide_lyase_gene_in_Nicotiana_benthamiana_using_a_viral_vector_system_ L2 - https://doi.org/10.1111/j.1467-7652.2010.00508.x DB - PRIME DP - Unbound Medicine ER -