Induction of secondary metabolism in grape cell cultures by jasmonates

2009 ◽  
Vol 36 (4) ◽  
pp. 323 ◽  
Author(s):  
Claudio D' Onofrio ◽  
Agnieszka Cox ◽  
Christopher Davies ◽  
Paul K. Boss
Keyword(s):  
Secondary Metabolites ◽  
Cell Cultures ◽  
Mass Spectra ◽  
Genetic Approach ◽  
Microarray Experiments ◽  
Expression Of Genes ◽  
Generation Times ◽  
Meja Treatment ◽  
Grape Cell ◽  
Stilbene Biosynthesis

The use of a genetic approach to study the biosynthetic pathways leading to the production of secondary metabolites in grapes is difficult given the long generation times and difficulty in transforming this species. In the present study, GC/MS and microarray experiments were used to identify compounds produced in grape cell cultures in response to jasmonates and to examine the expression of genes from pathways that produce these secondary metabolites. Methyl jasmonate (MeJA) and jasmonic acid (JA) treatments resulted in the production of at least 25 compounds with sesquiterpene-like mass spectra in the cell cultures. A significantly greater amount of proanthocyanidins was produced in the MeJA-treated cell cultures compared with controls and stilbene biosynthesis was induced in both MeJA- and JA-treated cells. Salicylic acid (SA) suppressed the MeJA-associated increase in sesquiterpenes and proanthocyanidins, but SA did not suppress the stilbene production induced by MeJA treatment. The mechanism by which jasmonates induced secondary metabolite production in cultured grape cells varied depending on the pathway. The increased production of proanthocyanidins and stilbenes was associated with the induction of all of the genes in associated biosynthesis pathways, including those involved in the production of phenylalanine, whereas increased sesquiterpene synthesis was linked to the induction of certain genes from relevant biosynthesis pathways.

Obtaining and Characterization of Alhagi persarum Boiss. et Buhse Callus Cell Cultures, Isoflavonoid Producers

2020 ◽  
Vol 36 (6) ◽  
pp. 35-48
Author(s):  
D.V. Коchkin ◽  
G.I. Sobolkovа ◽  
А.А. Fоmеnkov ◽  
R.А. Sidorov ◽  
А.М. Nоsоv
Keyword(s):  
Fatty Acids ◽  
Cell Culture ◽  
Liquid Chromatography ◽  
Secondary Metabolites ◽  
Cell Cultures ◽  
Mass Spectrometric ◽  
Mass Spectrometric Detection ◽  
Gas Liquid Chromatography ◽  
Callus Cell

The physiological characteristics of the callus cell cultures of Alhagi persarum Boiss et Buhse, a member of the legume family, widely used in folk medicine, have been studied. It was shown that the source of the explant was an important factor in the initiation of callusogenesis: more intense callusogenesis (almost 100%) was observed for explants from various organs of sterile seedlings, rather than intact plants (less than 30%). As a result, more than 20 lines of morphologically different callus cell cultures were obtained, and the growth parameters for the 5 most intensively growing lines were determined. The composition of fatty acids (FA) of total lipids and secondary metabolites in the most physiologically stable callus line Aр-207 was analyzed. Using capillary gas-liquid chromatography with mass spectrometric detection (GLC-MS), 19 individual C12--C24 FAs were identified, the main fraction of which were palmitic (~ 23%), stearic (~ 22%), linoleic (~ 14%) and α-linolenic (~ 33%) acids. The established atypical ratio of FAs (a simultaneous high content of both saturated FAs and polyunsaturated α-linolenic acid) is possibly due to the adaptation of cells to in vitro growth conditions. Phytochemical analysis of the secondary metabolites was carried out using ultra-performance liquid chromatography with electrospray ionization mass spectrometric detection (UPLC MS). Compounds belonging to different structural groups of isoflavones were found. Aglycones (calycosin, formononetin and afrormosin isomer), glucosides (formononetin glucoside), as well as esters of glucosides (malonylglycosides of calicosin, formononetin, afrormosin isomers, glycitein and genistein) were detected. These secondary metabolites are widespread in plants of the Fabaceae family; however, isoflavones are rare in representatives of the Alhagi genus. The presence of malonylated isoflavone glycosides in Alhagi spp. was shown for the first time. endemic plant species, Alhagi, in vitro cell culture, callus cell culture, isoflavones, fatty acids All studies were carried out using the equipment of the "Experimental Biotechnological Facility" and the "All-Russian Collection of Cell Cultures of Higher Plants" of IРР RAS. This work was supported by the Russian Foundation for Basic Research (RFBR), contract no.18-54-06021 (Az_a), and the Government of the Russian Federation, Megagrant Project no. 075-15-2019-1882.

scholarly journals The Influence of the Grapevine Bacterial and Fungal Endophytes on Biomass Accumulation and Stilbene Production by the In Vitro Cultivated Cells of Vitis amurensis Rupr.

Plants ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 1276
Author(s):  
Olga A. Aleynova ◽  
Andrey R. Suprun ◽  
Nikolay N. Nityagovsky ◽  
Alexandra S. Dubrovina ◽  
Konstantin V. Kiselev
Keyword(s):  
Cell Culture ◽  
Cell Suspension ◽  
Endophytic Fungi ◽  
Endophytic Bacteria ◽  
Fungal Endophytes ◽  
Biomass Accumulation ◽  
Vitis Amurensis ◽  
Grape Cell ◽  
Stilbene Biosynthesis

Plant endophytes are known to alter the profile of secondary metabolites in plant hosts. In this study, we identified the main bacterial and fungal representatives of the wild grape Vitis amurensis Rupr. microbiome and investigated a cocultivation effect of the 14 endophytes and the V. amurensis cell suspension on biomass accumulation and stilbene biosynthesis. The cocultivation of the V. amurensis cell culture with the bacteria Agrobacterium sp., Bacillus sp., and Curtobacterium sp. for 2 weeks did not significantly affect the accumulation of cell culture fresh biomass. However, it was significantly inhibited by the bacteria Erwinia sp., Pantoea sp., Pseudomonas sp., and Xanthomonas sp. and fungi Alternaria sp., Biscogniauxia sp., Cladosporium sp., Didymella sp. 2, and Fusarium sp. Cocultivation of the grapevine cell suspension with the fungi Didymella sp. 1 and Trichoderma sp. resulted in cell death. The addition of endophytic bacteria increased the total stilbene content by 2.2–5.3 times, while the addition of endophytic fungi was more effective in inducing stilbene accumulation by 2.6–16.3 times. The highest content of stilbenes in the grapevine cells cocultured with endophytic fungi was 13.63 and 13.76 mg/g of the cell dry weight (DW) after cultivation with Biscogniauxia sp. and Didymella sp. 2, respectively. The highest content of stilbenes in the grapevine cells cocultured with endophytic bacteria was 4.49 mg/g DW after cultivation with Xanthomonas sp. The increase in stilbene production was due to a significant activation of phenylalanine ammonia lyase (PAL) and stilbene synthase (STS) gene expression. We also analyzed the sensitivity of the selected endophytes to eight antibiotics, fluconazole, and trans-resveratrol. The endophytic bacteria were sensitive to gentamicin and kanamycin, while all selected fungal strains were resistant to fluconazole with the exception of Cladosporium sp. All endophytes were tolerant of trans-resveratrol. This study showed that grape endophytes stimulate the production of stilbenes in grape cell suspension, which could further contribute to the generation of a new stimulator of stilbene biosynthesis in grapevine or grape cell cultures.

Production of Secondary Metabolites in Cell Cultures of Some Terpenoid-Indole Alkaloids Producing Plants

1987 ◽  
pp. 485-494
Author(s):  
R. Verpoorte ◽  
R. Wijnsma ◽  
P. A. A. Harkes ◽  
H. J. G. ten Hoopen ◽  
J. J. Meijer ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Cell Cultures ◽  
Indole Alkaloids ◽  
Terpenoid Indole Alkaloids

Secondary Metabolites in Cell Cultures of Tabernaemontana Species

1989 ◽  
pp. 138-148
Author(s):  
R. Verpoorte ◽  
R. van der Heijden ◽  
J. Schripsema ◽  
M. Sierra ◽  
E. J. M. Pennings ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Cell Cultures

scholarly journals Advances in targeting and heterologous expression of genes involved in the synthesis of fungal secondary metabolites

RSC Advances ◽  
2019 ◽  
Vol 9 (60) ◽  
pp. 35124-35134 ◽  
Author(s):  
Yun-Ming Qiao ◽  
Rui-Lin Yu ◽  
Ping Zhu
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Secondary Metabolites ◽  
Heterologous Expression ◽  
Gene Targeting ◽  
Fungal Genome ◽  
P Gene ◽  
Expression Of Genes ◽  
Foreign Dna ◽  
Fungal Secondary Metabolites

Gene targeting involves integration of foreign DNA into the fungal genome by several strategies including Agrobacterium tumefaciens-mediated transformation (ATMT).

A 1-deoxy-d-xylulose 5-phosphate reductoisomerase gene probably involved in the synthesis of terpenoids in Chrysanthemum indicum var. aromaticum

2018 ◽  
Vol 98 (6) ◽  
pp. 1254-1264 ◽  
Author(s):  
Wenjie Gao ◽  
Xiang Wang ◽  
Nuananong Purente ◽  
Luqman Muhammad ◽  
Yunwei Zhou ◽  
...  
Keyword(s):  
Secondary Metabolites ◽  
Chemical Properties ◽  
Bioinformatic Analysis ◽  
Protein Product ◽  
Induction Effect ◽  
Tertiary Structures ◽  
Chrysanthemum Indicum ◽  
Meja Treatment ◽  
Key Enzyme ◽  
Physical And Chemical

Terpenoids are important secondary metabolites in plants. The synthesis of terpenoids involves a key enzyme, 1-deoxy-d-xylulose 5-phosphate reductoisomerase (DXR). In an aromatic ornamental plant, Chrysanthemum indicum var. aromaticum, terpenoids were found to be an important component in leaf secretions. To study the regulatory mechanism of terpenoid synthesis in C. indicum var. aromaticum, we cloned the DXR gene from C. indicum var. aromaticum, named CiDXR, based on transcriptome data. CiDXR is 1419 bp, encoding a polypeptide chain of 472 amino acids. Comparative and bioinformatic analysis showed that CiDXR was highly similar to DXRs from other plant species. The protein product of CiDXR was analyzed for physical and chemical properties, and secondary and tertiary structures. Methyl jasmonate (MeJA) is an effective exogenous elicitor that can impact the synthesis of secondary metabolites by regulating the expression of a series of specific key enzymes. In our study, MeJA treatment was used to study the relationship between CiDXR expression and terpenoid metabolism. The expression pattern of CiDXR was analyzed by quantitative real-time polymerase chain reaction. Leaves expressed the highest level of CiDXR mRNA. In addition, CiDXR expression was found to be activated by MeJA, up to 20-fold at 24 h post induction. The MeJA treatment significantly affected the relative contents of the secretion products, and the induction effect was maximal at 24 h, consistent with the expression of CiDXR. This study shows that CiDXR may play a key regulatory role in the control of terpenoid synthesis in C. indicum var. aromaticum.

scholarly journals The influence of downy mildew tolerance of grape varieties on the biosynthesis of stilbenes in callus as potential sources of bioactive substances

2020 ◽  
Vol 25 ◽  
pp. 02013
Author(s):  
Eugene Lutsky ◽  
Svyatoslav Fedorovich ◽  
Vadim Vyalkov ◽  
Maria Sundyreva
Keyword(s):  
Downy Mildew ◽  
Callus Culture ◽  
Practical Interest ◽  
Stilbene Synthase ◽  
Necessary Condition ◽  
Bioactive Substances ◽  
Expression Of Genes ◽  
Grape Varieties ◽  
Stilbene Biosynthesis ◽  
Effective Source

Stilbenes represent a considerable practical interest in relation to their benifits to the human health. Callus culture of the grape, which is a natural producer of stilbenes, can serve as an effective source of these compounds. Grape’s resistance to biotic environmental stress conditions is associated with the synthesis and conversion of stilbenes, therefore the aim of the current study was to determine an interrelation between the tolerance of grape varieties to downy mildew and stilbene biosynthesis in the callus culture. Obtained results showed that there was no link between variety’s tolerance to downy mildew and content of stilbenes, chalcones and phenoloxydising enzymes in callus. Presence of the stress impact is a necessary condition for the initiation of the variety-specific synthesis of stilbenes. Callus of varieties, in which stilbenes content was higher, was characterized by a lowered expression of the chalcone synthase and chalcone isomerase relative to expression of genes of the stilbene synthase and phenylalanine ammonia liase, which confirms an interrelation between reduced competition for the substrate between two biosynthesis branches of chalcones and stilbenes with production of the latter.

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