Cargando…

Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage

BACKGROUND: Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an al...

Descripción completa

Detalles Bibliográficos
Autores principales: Palama, Tony L, Menard, Patrice, Fock, Isabelle, Choi, Young H, Bourdon, Emmanuel, Govinden-Soulange, Joyce, Bahut, Muriel, Payet, Bertrand, Verpoorte, Robert, Kodja, Hippolyte
Formato: Texto
Lenguaje:English
Publicado: BioMed Central 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095354/
https://www.ncbi.nlm.nih.gov/pubmed/20444255
http://dx.doi.org/10.1186/1471-2229-10-82
_version_ 1782203637759475712
author Palama, Tony L
Menard, Patrice
Fock, Isabelle
Choi, Young H
Bourdon, Emmanuel
Govinden-Soulange, Joyce
Bahut, Muriel
Payet, Bertrand
Verpoorte, Robert
Kodja, Hippolyte
author_facet Palama, Tony L
Menard, Patrice
Fock, Isabelle
Choi, Young H
Bourdon, Emmanuel
Govinden-Soulange, Joyce
Bahut, Muriel
Payet, Bertrand
Verpoorte, Robert
Kodja, Hippolyte
author_sort Palama, Tony L
collection PubMed
description BACKGROUND: Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an alternative to minimize damage to mother plants. Because mass production of V. planifolia through indirect shoot differentiation from callus culture is rare and may be a successful use of in vitro techniques for producing somaclonal variants, we have established a novel protocol for the regeneration of vanilla plants and investigated the initial biochemical and molecular mechanisms that trigger shoot organogenesis from embryogenic/organogenic callus. RESULTS: For embryogenic callus induction, seeds obtained from 7-month-old green pods of V. planifolia were inoculated on MS basal medium (BM) containing TDZ (0.5 mg l(-1)). Germination of unorganized mass callus such as protocorm -like structure (PLS) arising from each seed has been observed. The primary embryogenic calli have been formed after transferring on BM containing IAA (0.5 mg l(-1)) and TDZ (0.5 mg l(-1)). These calli were maintained by subculturing on BM containing IAA (0.5 mg l(-1)) and TDZ (0.3 mg l(-1)) during 6 months and formed embryogenic/organogenic calli. Histological analysis showed that shoot organogenesis was induced between 15 and 20 days after embryogenic/organogenic calli were transferred onto MS basal medium with NAA (0.5 mg l(-1)). By associating proteomics and metabolomics analyses, the biochemical and molecular markers responsible for shoot induction have been studied in 15-day-old calli at the stage where no differentiating part was visible on organogenic calli. Two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization time-of-flight-tandem mass spectrometry (MALDI-TOF-TOF-MS) analysis revealed that 15 protein spots are significantly expressed (P < 0.05) at earlier stages of shoot differentiation. The majority of these proteins are involved in amino acid-protein metabolism and photosynthetic activity. In accordance with proteomic analysis, metabolic profiling using 1D and 2D NMR techniques showed the importance of numerous compounds related with sugar mobilization and nitrogen metabolism. NMR analysis techniques also allowed the identification of some secondary metabolites such as phenolic compounds whose accumulation was enhanced during shoot differentiation. CONCLUSION: The subculture of embryogenic/organogenic calli onto shoot differentiation medium triggers the stimulation of cell metabolism principally at three levels namely (i) initiation of photosynthesis, glycolysis and phenolic compounds synthesis; (ii) amino acid - protein synthesis, and protein stabilization; (iii) sugar degradation. These biochemical mechanisms associated with the initiation of shoot formation during protocorm - like body (PLB) organogenesis could be coordinated by the removal of TDZ in callus maintenance medium. These results might contribute to elucidate the complex mechanism that leads to vanilla callus differentiation and subsequent shoot formation into PLB organogenesis. Moreover, our results highlight an early intermediate metabolic event in vanillin biosynthetic pathway with respect to secondary metabolism. Indeed, for the first time in vanilla tissue culture, phenolic compounds such as glucoside A and glucoside B were identified. The degradation of these compounds in specialized tissue (i.e. young green beans) probably contributes to the biosynthesis of glucovanillin, the parent compound of vanillin.
format Text
id pubmed-3095354
institution National Center for Biotechnology Information
language English
publishDate 2010
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-30953542011-05-17 Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage Palama, Tony L Menard, Patrice Fock, Isabelle Choi, Young H Bourdon, Emmanuel Govinden-Soulange, Joyce Bahut, Muriel Payet, Bertrand Verpoorte, Robert Kodja, Hippolyte BMC Plant Biol Research Article BACKGROUND: Vanilla planifolia is an important Orchid commercially cultivated for the production of natural vanilla flavour. Vanilla plants are conventionally propagated by stem cuttings and thus causing injury to the mother plants. Regeneration and in vitro mass multiplication are proposed as an alternative to minimize damage to mother plants. Because mass production of V. planifolia through indirect shoot differentiation from callus culture is rare and may be a successful use of in vitro techniques for producing somaclonal variants, we have established a novel protocol for the regeneration of vanilla plants and investigated the initial biochemical and molecular mechanisms that trigger shoot organogenesis from embryogenic/organogenic callus. RESULTS: For embryogenic callus induction, seeds obtained from 7-month-old green pods of V. planifolia were inoculated on MS basal medium (BM) containing TDZ (0.5 mg l(-1)). Germination of unorganized mass callus such as protocorm -like structure (PLS) arising from each seed has been observed. The primary embryogenic calli have been formed after transferring on BM containing IAA (0.5 mg l(-1)) and TDZ (0.5 mg l(-1)). These calli were maintained by subculturing on BM containing IAA (0.5 mg l(-1)) and TDZ (0.3 mg l(-1)) during 6 months and formed embryogenic/organogenic calli. Histological analysis showed that shoot organogenesis was induced between 15 and 20 days after embryogenic/organogenic calli were transferred onto MS basal medium with NAA (0.5 mg l(-1)). By associating proteomics and metabolomics analyses, the biochemical and molecular markers responsible for shoot induction have been studied in 15-day-old calli at the stage where no differentiating part was visible on organogenic calli. Two-dimensional electrophoresis followed by matrix-assisted laser desorption ionization time-of-flight-tandem mass spectrometry (MALDI-TOF-TOF-MS) analysis revealed that 15 protein spots are significantly expressed (P < 0.05) at earlier stages of shoot differentiation. The majority of these proteins are involved in amino acid-protein metabolism and photosynthetic activity. In accordance with proteomic analysis, metabolic profiling using 1D and 2D NMR techniques showed the importance of numerous compounds related with sugar mobilization and nitrogen metabolism. NMR analysis techniques also allowed the identification of some secondary metabolites such as phenolic compounds whose accumulation was enhanced during shoot differentiation. CONCLUSION: The subculture of embryogenic/organogenic calli onto shoot differentiation medium triggers the stimulation of cell metabolism principally at three levels namely (i) initiation of photosynthesis, glycolysis and phenolic compounds synthesis; (ii) amino acid - protein synthesis, and protein stabilization; (iii) sugar degradation. These biochemical mechanisms associated with the initiation of shoot formation during protocorm - like body (PLB) organogenesis could be coordinated by the removal of TDZ in callus maintenance medium. These results might contribute to elucidate the complex mechanism that leads to vanilla callus differentiation and subsequent shoot formation into PLB organogenesis. Moreover, our results highlight an early intermediate metabolic event in vanillin biosynthetic pathway with respect to secondary metabolism. Indeed, for the first time in vanilla tissue culture, phenolic compounds such as glucoside A and glucoside B were identified. The degradation of these compounds in specialized tissue (i.e. young green beans) probably contributes to the biosynthesis of glucovanillin, the parent compound of vanillin. BioMed Central 2010-05-05 /pmc/articles/PMC3095354/ /pubmed/20444255 http://dx.doi.org/10.1186/1471-2229-10-82 Text en Copyright ©2010 Palama et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research Article
Palama, Tony L
Menard, Patrice
Fock, Isabelle
Choi, Young H
Bourdon, Emmanuel
Govinden-Soulange, Joyce
Bahut, Muriel
Payet, Bertrand
Verpoorte, Robert
Kodja, Hippolyte
Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title_full Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title_fullStr Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title_full_unstemmed Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title_short Shoot differentiation from protocorm callus cultures of Vanilla planifolia (Orchidaceae): proteomic and metabolic responses at early stage
title_sort shoot differentiation from protocorm callus cultures of vanilla planifolia (orchidaceae): proteomic and metabolic responses at early stage
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3095354/
https://www.ncbi.nlm.nih.gov/pubmed/20444255
http://dx.doi.org/10.1186/1471-2229-10-82
work_keys_str_mv AT palamatonyl shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT menardpatrice shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT fockisabelle shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT choiyoungh shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT bourdonemmanuel shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT govindensoulangejoyce shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT bahutmuriel shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT payetbertrand shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT verpoorterobert shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage
AT kodjahippolyte shootdifferentiationfromprotocormcallusculturesofvanillaplanifoliaorchidaceaeproteomicandmetabolicresponsesatearlystage