Interlinking showy traits: co-engineering of scent and colour biosynthesis in flowers
Michal Moyal Ben Zvi
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorFlorence Negre-Zakharov
Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA
Present address: Department of Plant Sciences, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA
Search for more papers by this authorTania Masci
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorMarianna Ovadis
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorElena Shklarman
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorHagit Ben-Meir
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorTzvi Tzfira
Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI 48109, USA
Search for more papers by this authorNatalia Dudareva
Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA
Search for more papers by this authorCorresponding Author
Alexander Vainstein
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
* Correspondence (fax 972-8-9489091; e-mail [email protected])Search for more papers by this authorMichal Moyal Ben Zvi
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorFlorence Negre-Zakharov
Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA
Present address: Department of Plant Sciences, University of California-Davis, One Shields Avenue, Davis, CA 95616, USA
Search for more papers by this authorTania Masci
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorMarianna Ovadis
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorElena Shklarman
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorHagit Ben-Meir
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
Search for more papers by this authorTzvi Tzfira
Department of Molecular, Cellular and Developmental Biology, The University of Michigan, Ann Arbor, MI 48109, USA
Search for more papers by this authorNatalia Dudareva
Department of Horticulture and Landscape Architecture, Purdue University, West Lafayette, IN 47907, USA
Search for more papers by this authorCorresponding Author
Alexander Vainstein
The Institute of Plant Sciences and Genetics in Agriculture, Faculty of Agricultural, Food and Environmental Quality Sciences, The Hebrew University of Jerusalem, PO Box 12, Rehovot 76100, Israel
* Correspondence (fax 972-8-9489091; e-mail [email protected])Search for more papers by this authorSummary
The phenylpropanoid pathway gives rise to metabolites that determine floral colour and fragrance. These metabolites are one of the main means used by plants to attract pollinators, thereby ensuring plant survival. A lack of knowledge about factors regulating scent production has prevented the successful enhancement of volatile phenylpropanoid production in flowers. In this study, the Production of Anthocyanin Pigment1 (Pap1) Myb transcription factor from Arabidopsis thaliana, known to regulate the production of non-volatile phenylpropanoids, including anthocyanins, was stably introduced into Petunia hybrida. In addition to an increase in pigmentation, Pap1-transgenic petunia flowers demonstrated an increase of up to tenfold in the production of volatile phenylpropanoid/benzenoid compounds. The dramatic increase in volatile production corresponded to the native nocturnal rhythms of volatile production in petunia. The application of phenylalanine to Pap1-transgenic flowers led to an increase in the otherwise negligible levels of volatiles emitted during the day to nocturnal levels. On the basis of gene expression profiling and the levels of pathway intermediates, it is proposed that both increased metabolic flux and transcriptional activation of scent and colour genes underlie the enhancement of petunia flower colour and scent production by Pap1. The co-ordinated regulation of metabolic steps within or between pathways involved in vital plant functions, as shown here for two showy traits determining plant–pollinator interactions, provides a clear advantage for plant survival. The use of a regulatory factor that activates scent production creates a new biotechnological strategy for the metabolic architecture of fragrance, leading to the creation of novel genetic variability for breeding purposes.
Supporting Information
Figure S1 Methylbenzoate production in flowers of control and Production of Anthocyanin Pigment1 (Pap1)-transgenic petunia lines. (a) Levels of methylbenzoate emitted from flowers of control (C) and Pap1-transgenic (P1-3, P1-2 and P1-7) lines were determined using the dynamic headspace technique. Analysis was conducted for 12 h during the first night post-anthesis. Columns represent the mean values of independent experiments (n = 3). Standard errors are indicated by vertical bars. Data were subjected to one-way analysis of variance (ANOVA), and no significant effect (P < 0.05) of genotype was revealed. (b) RNA-blot analysis of S-adenosyl-L-methionine:benzoic acid/salicylic acid carboxyl methyltransferase (Bsmt) levels in limbs [first (1) and second (2) days post-anthesis] of control (C) and Pap1-transgenic line 2 (P). The blot was rehybridized with an 18S rRNA probe to ensure equal loading of samples.
Figure S2 Developmental profiles of benzaldehyde emission in control and Production of Anthocyanin Pigment1 (Pap1)-transgenic petunia flowers. Levels of benzaldehyde emission from control (C) and Pap1-transgenic line 2 (P) flowers at different developmental stages were determined using the dynamic headspace technique. Analysis was conducted for 12 h using flowers during the first, second and third night post-anthesis (1, 2 and 3, respectively). Each point represents the mean values of independent experiments (n = 3). Standard errors are indicated by vertical bars. The significance of the differences in benzaldehyde emission between Pap1-transgenic line 2 and control non-transgenic flowers, at different developmental stages, was calculated using Tukey's all pairwise multiple comparison procedure following two-way analysis of variance (ANOVA). Values significantly different from control flowers are indicated by asterisks (P < 0.0001).
Figure S3 Benzaldehyde production during a day/night cycle in control and Production of Anthocyanin Pigment1 (Pap1)-transgenic petunia flowers. (a) Levels of benzaldehyde emission from control (C) and Pap1-transgenic line 2 (P) flowers 2 days post-anthesis were determined using the dynamic headspace technique. Headspace collections were performed during the day for 12 h under light conditions (06.00–18.00 h) and during the night for 12 h under dark conditions (18.00–06.00 h). (b) Determination of internal pools of benzaldehyde. Free and total (following glucosidase treatment) pools of benzaldehyde were assayed in limbs collected during the day (11.00 h) and night (23.00 h) using extraction with hexane. Columns represent the mean values of independent experiments (n = 3). Standard errors are indicated by vertical bars. The significance of the differences in the levels of benzaldehyde emission (a) and internal pool (b) between Pap1-transgenic line 2 and control non-transgenic flowers during the day/night cycle was calculated using Tukey's all pairwise multiple comparison procedure following two-way and multi-factor analysis of variance (ANOVA), respectively. Values with different letters are significantly different (P < 0.05 and P < 0.0001 in a and b, respectively).
Figure S4 Pools of cinnamic (a) and coumaric (b) acids in control and Production of Anthocyanin Pigment1 (Pap1)-transgenic petunia flowers. The levels of cinnamic and coumaric acids in the limbs of control (C) and Pap1-transgenic line 2 (P) petunia were analysed during the day (11.00 h) and at night (23.00 h) on the first day post-anthesis. Columns represent the mean values of independent experiments (n = 3). Standard errors are indicated by vertical bars. Data were subjected to two-way analysis of variance (ANOVA), and no significant effect (P < 0.05) of treatments was revealed.
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