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Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus lupulus L.)

J. Matousek, T. Kocabek, J. Patzak, J. Stehlik, Z. Fussy, K. Krofta, Arne Heyerick UGent, Isabel Roldàn-Ruiz UGent, Lina Maloukh UGent and Denis De Keukeleire UGent (2010) Journal of Agricultural and Food Chemistry. 58(2). p.902-912
abstract
Hop (Humulus lupulus L.), the essential source of beer flavor is of interest from a medicinal perspective in view of its high content in health-beneficial terpenophenolics including prenylflavonoids. The dissection of biosynthetic pathway(s) of these compounds in lupulin glands, as well as its regulation by transcription factors (TFs), is important for efficient biotech no logical manipulation of the hop metabolome. TFs of the bZIP class were preselected from the hop transcriptome using a cDNA-AFLP approach and cloned from a cDNA library based on glandular tissue-enriched hop cones. The cloned TFs HlbZIP1A and HlbZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively, and both are similar to the group A3 bZIP TFs according to the composition of characteristic domains. While HlbZIP1A is rather neutral (pl 6.42), HlbZIP2 is strongly basic (pl 8,51). A truncated variant of HlbZIP1 (HlbZIP1B), which is strongly basic but lacks the leucine zipper domain, has also been cloned from hop. Similar to the previously cloned HIMyb3 from hop, both bZIP TFs show a highly specific expression in lupulin glands, although low expression was observed also in other tissues including roots and immature pollen. Comparative functional analyses of HbZip1A, HlbZip2, and subvariants of HIMyb3 were performed in a transient expression system using Nicotiana benthamiana leaf coinfiltration with Agrobacterium tumefaciens strains bearing hop TFs and selected promoters fused to the GUS reference gene. both hop bZIP TFs and HIMyb3 mainly activated the promoters of chalcone synthase chs_H1 and the,newly cloned O-methyl transferase 1 genes, while the response of the valerophenone synthase promoter to the cloned hop TFs was very low. These analyses also showed that the cloned bZIP TFs are not strictly G-box-specific. HPLC analysis of secondary metabolites in infiltrated Petunia hybrida showed that both hop bZIP TFs interfere with the accumulation and the composition of flavonol glycosides, phenolic acids, and anthocyanins, suggesting the possibility of coregulating flavonoid biosynthetic pathways in hop glandular tissue.
Please use this url to cite or link to this publication:
author
organization
year
type
journalArticle (original)
publication status
published
subject
keyword
Petunia hybrida, secondary metabolites, Nicotiana benthamiana, transient expression, hop cDNA library screening, Transcriptional factors, cDNA-AFLP analysis
journal title
Journal of Agricultural and Food Chemistry
J. Agric. Food Chem.
volume
58
issue
2
pages
902 - 912
Web of Science type
Article
Web of Science id
000273671900032
JCR category
AGRICULTURE, MULTIDISCIPLINARY
JCR impact factor
2.816 (2010)
JCR rank
2/55 (2010)
JCR quartile
1 (2010)
ISSN
0021-8561
DOI
10.1021/jf9043106
language
English
UGent publication?
yes
classification
A1
copyright statement
I have transferred the copyright for this publication to the publisher
id
872953
handle
http://hdl.handle.net/1854/LU-872953
date created
2010-02-22 11:57:52
date last changed
2010-03-12 14:50:58
@article{872953,
  abstract     = {Hop (Humulus lupulus L.), the essential source of beer flavor is of interest from a medicinal perspective in view of its high content in health-beneficial terpenophenolics including prenylflavonoids. The dissection of biosynthetic pathway(s) of these compounds in lupulin glands, as well as its regulation by transcription factors (TFs), is important for efficient biotech no logical manipulation of the hop metabolome. TFs of the bZIP class were preselected from the hop transcriptome using a cDNA-AFLP approach and cloned from a cDNA library based on glandular tissue-enriched hop cones. The cloned TFs HlbZIP1A and HlbZIP2 have predicted molecular masses of 27.4 and 34.2 kDa, respectively, and both are similar to the group A3 bZIP TFs according to the composition of characteristic domains. While HlbZIP1A is rather neutral (pl 6.42), HlbZIP2 is strongly basic (pl 8,51). A truncated variant of HlbZIP1 (HlbZIP1B), which is strongly basic but lacks the leucine zipper domain, has also been cloned from hop. Similar to the previously cloned HIMyb3 from hop, both bZIP TFs show a highly specific expression in lupulin glands, although low expression was observed also in other tissues including roots and immature pollen. Comparative functional analyses of HbZip1A, HlbZip2, and subvariants of HIMyb3 were performed in a transient expression system using Nicotiana benthamiana leaf coinfiltration with Agrobacterium tumefaciens strains bearing hop TFs and selected promoters fused to the GUS reference gene. both hop bZIP TFs and HIMyb3 mainly activated the promoters of chalcone synthase chs\_H1 and the,newly cloned O-methyl transferase 1 genes, while the response of the valerophenone synthase promoter to the cloned hop TFs was very low. These analyses also showed that the cloned bZIP TFs are not strictly G-box-specific. HPLC analysis of secondary metabolites in infiltrated Petunia hybrida showed that both hop bZIP TFs interfere with the accumulation and the composition of flavonol glycosides, phenolic acids, and anthocyanins, suggesting the possibility of coregulating flavonoid biosynthetic pathways in hop glandular tissue.},
  author       = {Matousek, J. and Kocabek, T. and Patzak, J. and Stehlik, J. and Fussy, Z. and Krofta, K. and Heyerick, Arne and Rold{\`a}n-Ruiz, Isabel and Maloukh, Lina and De Keukeleire, Denis},
  issn         = {0021-8561},
  journal      = {Journal of Agricultural and Food Chemistry},
  keyword      = {Petunia hybrida,secondary metabolites,Nicotiana benthamiana,transient expression,hop cDNA library screening,Transcriptional factors,cDNA-AFLP analysis},
  language     = {eng},
  number       = {2},
  pages        = {902--912},
  title        = {Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus lupulus L.)},
  url          = {http://dx.doi.org/10.1021/jf9043106},
  volume       = {58},
  year         = {2010},
}

Chicago
Matousek, J., T. Kocabek, J. Patzak, J. Stehlik, Z. Fussy, K. Krofta, Arne Heyerick, Isabel Roldàn-Ruiz, Lina Maloukh, and Denis De Keukeleire. 2010. “Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus Lupulus L.).” Journal of Agricultural and Food Chemistry 58 (2): 902–912.
APA
Matousek, J., Kocabek, T., Patzak, J., Stehlik, J., Fussy, Z., Krofta, K., Heyerick, A., et al. (2010). Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus lupulus L.). Journal of Agricultural and Food Chemistry, 58(2), 902–912.
Vancouver
1.
Matousek J, Kocabek T, Patzak J, Stehlik J, Fussy Z, Krofta K, et al. Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus lupulus L.). Journal of Agricultural and Food Chemistry. 2010;58(2):902–12.
MLA
Matousek, J., T. Kocabek, J. Patzak, et al. “Cloning and Molecular Analysis of HlbZip1 and HlbZip2 Transcription Factors Putatively Involved in the Regulation of the Lupulin Metabolome in Hop (Humulus Lupulus L.).” Journal of Agricultural and Food Chemistry 58.2 (2010): 902–912. Print.