CorrectionsAGRICULTURAL SCIENCES. For the article ''Characterization of capsaicin synthase and identification of its gene (csy1) for pungency factor capsaicin in pepper (Capsicum sp.),'' by B. C. Narasimha
Capsaicin, a pungency factor alkaloid of Capsicum fruits, is biosynthesized by enzymatic condensation of vanillylamine, a phenyl propanoid intermediate, and 8-methyl-nonenoic acid, a fatty acid derivative from the leucine/valine pathway by capsaicin synthase. Biotic elicitors, such as aqueous mycelial extracts of Rhizopus oligosporus and Aspergillus niger, and abiotic elicitors, such as salicylic acid and methyl jasmonate, were incorporated to cell-suspension cultures of Capsicum frutescens. The maximum elicitation of capsaicinoids to the extent of 6-folds was noticed when mycelial extracts of R. oligosporus were incorporated to cell-suspension cultures. The phenyl propanoid intermediates were elicited to the extent of 3-4-fold, whereas the levels of 8-methyl-nonanoic acid increased 6-fold in R. oligosporus mycelial-extract-treated cultures. The enzymatic assays of caffeic acid O-methyl transferases, keto acyl synthase, and capsaicin synthase were performed in elicited and nonelicited cell-suspension cultures. These enzymes were overexpressed in R. oligosporus mycelial-extract-treated suspension cultures, which was also confirmed by reverse transcriptase polymerase chain reaction studies. This is the first paper on the overexpression of 8-methyl-nonanoic acid leading to capsaicin biosynthesis using elicitors.
Direct shoot bud induction and plant regeneration was achieved in Capsicum frutescens var. KTOC. Aseptically grown seedling explants devoid of roots, apical meristem and cotyledons were inoculated in an inverted position in medium comprising of Murashige and Skoog (Physiol Plant 15:472-497, 1962) basal medium supplemented with 2-(N-morpholine) ethanesulphonic acid buffer along with 2.28 lM indole-3-acetic acid, 10 lM silver nitrate and either of 13.31-89.77 lM benzyl adenine (BA), 9.29-23.23 lM kinetin, 0.91-9.12 lM zeatin, 2.46-9.84 lM 2-isopentenyl adenine. Profuse shoot bud induction was observed only in explants grown on a media supplemented with BA (26.63 lM) as a cytokinin source and 19.4 ± 4.2 shoot buds per explant was obtained in inverted mode under continuous light. Incorporation of polyamine inhibitors in the culture medium completely inhibited shoothoot bud induction. Incorporation of exogenous polyamines improved the induction of shoot buds under 24 h photoperiod. These buds were elongated in MS medium containing 2.8 lM gibberellic acid. Transfer of these shoots to hormone-free MS medium resulted in rooting and rooted plants were transferred to fields. This protocol can be efficiently used for mass propagation and presumably also for regeneration of genetically transformed C. frutescens.
Capsaicin is a bioactive molecule synthesized by enzymatic (putative capsaicin synthase) condensation of vanillylamine, a phenyl propanoid intermediate with 8-methyl-nonenoic acid, a fatty acid derivative from leucine/valine pathway. Analysis of levels of 8-methyl-nonenoic acid and phenyl propanoid intermediates in high, medium, and low pungent Capsicum genotypes revealed that the 8-methyl-nonenoic acid pool plays a crucial role in determining the efficacy of capsaicin levels. Cerulenin-mediated inhibition of 8-methyl-nonenoic acid synthesis decreased the capsaicin biosynthesis in Capsicum cell suspension cultures. Similarly amino oxy acetate inhibited vanillylamine synthesis but failed to reduce capsaicin production. The mRNA transcript analysis of keto acyl synthase (KAS), a crucial enzyme involved in 8-methyl-nonenoic acid and an amino transferase (AMT), involved in vanillylamine biosynthesis was studied. The mRNA transcript analysis revealed the progressive developmental expression of KAS gene in the placenta during the ontogeny of the fruit, whereas AMT transcripts levels did not show significant differences. Hence, the study demonstrates the influence of 8-methyl-nonenoic acid and its possible regulatory role in capsaicin biosynthesis in Capsicum spp.
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