In vitro plantlet regeneration from cotyledon segments of Capsicum chinense Jacq. cv. Naga King Chili, and determination of capsaicin content in fruits of in vitro propagated plants by High Performance Liquid Chromatography (original) (raw)
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IN VITRO PRODUCTION OF CAPSAICIN THROUGH PLANT TISSUE CULTURE
Journal of Phytology, 2017
Capsaicin, a secondary metabolite produced in capsicum, is in high demand in pharmaceutical industry because of its various medicinal properties. Currently, the supply of capsaicin depends upon its extraction from capsicum fruits. This limits the production of capsaicin as it depends upon agricultural produce. The current review has compiled information from various literature published on chemistry and importance of capsaicin along with its method of production. It also reviews the process of in vitro production of capsaicin through plant tissue culture, strategies of increasing capsaicin accumulation and its advantages over extraction from fruits and artificial synthesis.
Elicitation of cell suspension cultures of Capsicum assamicum (Bhut Jolokia) for enhancement of capsaicin content was tried using different elicitors such as cellulase, vanillin, methyl jasmonate, salicylic acid and sinapic acid in different concentrations for 24, 48 and 72 hours. Cell suspension culture was established in B5 media supplemented with 3.5 mM 2,4-D (2,4-diphenoxyacetic acid) and 1.1 mM Kin and elicitors were introduced at the end of exponential phase. All the elicitors, except methyl jasmonate, led to significant increase in production of capsaicin. Sinapic acid, when added in 22 µM concentration and incubated for 24 hours, led to highest capsaicin accumulation of 0.5% (5068 µg/g) which was highest among all the treatments.
The effect of different concentrations of cellulase on the production of capsaicin in freely suspended cell and immobilized cell cultures of Kahramanmaraş pepper seeds (Capsicum annuum L.) were studied. Calluses were obtained from in vitro germinated hypocotyl explants of pepper seedlings and cell suspensions were prepared from these calluses. Immobilized cell suspension cultures with calcium alginate and free cell suspension cultures were obtained by using cell suspensions. Elicitor such as cellulase (5-30 µg/ml), was applied both for the free and immobilized cell suspensions and control group without elicitor was prepared. The concentration of capsaicin in freely suspended cells, immobilized cells and their filtrates were identified by HPLC after extraction with ethyl acetate. It was found that the immobilization process had an increasing effect on the capsaicin accumulation. The concentration of capsaicin in the immobilized cells for both control groups and elicitor added samples was higher than the free cells. In general, capsaicin concentration in the filtrate for free cells was higher than the immobilized cells. When all the cellulase and the sampling hours were compared, the highest capsaicin concentration for the immobilized cells was determined as 362,91 µg/g f.w. at the 24th hour for 30 µg/ml cellulase applied samples.
TURKISH JOURNAL OF AGRICULTURE AND FORESTRY
Introduction The term "vegetable" usually refers to the fresh edible portions of certain herbaceous plants' roots, stems, leaves, flowers, fruit, or seeds and these plant parts are either eaten fresh or prepared in a number of ways for nutritional purposes (Fadda et al., 2018; Galiana-Belaguer et al., 2018). Pepper (Capsicum spp.) is one of the warm-climate vegetables grown widely in both open fields and greenhouses worldwide (Sarafi et al., 2018). Turkey is one of the top three pepper-producing countries with over 2 million tons (http://faostat3.fao.org/). The germination and emergence of pepper seeds is often slow and nonuniform under normal as well as stressed conditions (Lorenz and Maynard, 1988; Chartzoulakis and Klapaki, 2000) Also, when compared with other vegetable seeds, its viable storage period is very short and seeds deteriorate quickly (Demir and Okcu, 2004; Khan et al., 2009; Yadav et al., 2011). Nongerminating seeds and nonuniform and unhealthy plants increase the cost of plants growing per area. In recent years, there has been widespread vegetablegrowing throughout Turkey that requires investment in establishing seedlings. Seedling quality is very important in species grown from seedlings because it is affected by the growth media, climatic conditions, and cultivar applications after planting, which affect the plant growth and yield (Sahin et al., 2002; Korkmaz et al., 2010; Rajjou et al., 2012). The aim of commercial seedling production is to obtain a high emergence rate as well as healthy and homogeneous seedlings in a short time period. There are some applications, called priming, that initiate the physiological process of germination with limited water intake under controlled conditions of seeds prior to sowing, to obtain high and homogeneous seed germination both from direct sowing in the field and seedling growth. This technique is used to improve the germination and emergence of some vegetable species (Bradford et al., 1990; Pill, 1995; Taylor et al., 1998). In works done on pepper where osmotic solutions of polyethylene glycol and K and Na salts are used, it has been shown that germination and emergence are generally improved, especially under biotic and abiotic stress conditions (Amjad et al., 2007; Korkmaz and Korkmaz, 2009; Yadav et al., 2011). Capsaicin, which is synthesized and accumulated in the placental tissues of the fruit, is the major pungent
2014
The effect of different concentrations of cellulase on the production of capsaicin in freely suspended cell and immobilized cell cultures of Kahramanmaraş pepper seeds (Capsicum annuum L.) were studied. Calluses were obtained from in vitro germinated hypocotyl explants of pepper seedlings and cell suspensions were prepared from these calluses. Immobilized cell suspension cultures with calcium alginate and free cell suspension cultures were obtained by using cell suspensions. Elicitor such as cellulase (5-30 μg/ml), was applied both for the free and immobilized cell suspensions and control group without elicitor was prepared. The concentration of capsaicin in freely suspended cells, immobilized cells and their filtrates were identified by HPLC after extraction with ethyl acetate. It was found that the immobilization process had an increasing effect on the capsaicin accumulation. The concentration of capsaicin in the immobilized cells for both control groups and elicitor added samples...
Regeneration potential of seedling explants of chilli ( Capsicum annuum )
African Journal of Biotechnology, 2009
A study was conducted with hypocotyl, cotyledon and shoot tip of chilli as explants for regeneration on MS medium supplemented with different concentrations and combinations of auxins and cytokinins. Regeneration potential was determined by two ways. One is regeneration of shoot via callus formation from hypocotyls and cotyledon explants; another was direct shoot regeneration from shoot tip explant. The highest callus was induced from hypocotyl in a combination of BAP (5.0 mgL-1) with NAA (0.1 mgL-1) and cotyledon in a combination of BAP (5.0 mgL-1) with IAA ((1.0 mgL-1). The callus induction as well as shoot initiation was higher in hypocotyls than cotyledon. Shoot tips regenerated into plantlets directly with sporadic small callus at the base. Shoot elongation was accelerated by using additional supplementation of GA3 and AgNO3. Regenerated shoots rooted best on the MS medium supplemented with 0.1 mg L-1 NAA + 0.05 mg L-1 IBA.
HPLC analysis of capsaicin content in sixteen Capsicum fruits from Nepal
Journal of Medicinal Food
Capsicum fruit, a popular spice as chili pepper, is an important ingredient of the formulations used in traditional medicines. Moreover, Capsicum fruit is listed as an official drug in several pharmacopoeias. Capsaicin, the most abundant component in Capsicum fruit, exhibits its therapeutic and adverse effects in a dose-dependent manner. Therefore, the known capsaicin content is the prerequisite for optimizing any formulation based on Capsicum fruit as a crude drug. We studied 16 samples of Capsicum fruits grown at different altitudes in Nepal and determined their capsaicin content by highperformance liquid chromatography. The capsaicin content was found to range from 2.19 to 19.73 mg=g of dry weight of Capsicum fruits. Capsaicin content in pericarp was found to be higher than in seeds. No correlation was found between the shape or size of the fruits and its capsaicin content. Our findings indicate that many of the formulations prepared from Capsicum fruit, even as described in pharmacopoeias, may vary in their strength, therapeutic activity, and possible side effects if the capsaicin content in Capsicum fruit is not standardized.
Nusantara bioscience, 2022
Land scarcity for cropping at Java Island is a challenge for scientists to look for alternative cropping land. The use of saline land for cropping needs to have further discussed. Red pepper (Capsicum annuum L.) can be used as a plant model because, aside from being used as a vegetable, it is also used as natural medicine because of its secondary metabolite, capsaicin. A harsh environment could induce changes in the primary metabolism, which leads to secondary metabolite decomposition. For example, plants respond to stress, such as salt stress, by synthesizing flavonoids and phenolic acid as defense systems to reduce damage. However, the total sugar level and organic acids are decreased. This research aimed to study the fruit development and capsaicin content of hot pepper grown on various coastal soil sand to know whether or not different growth medium affects the size of each part of the fruit. The design of this research was a Completely Randomized Block Design (CRBD). In this research, five different salinity mediums were used, they were A. 15.20 dS/m, B. 5.70 dS/m, C.1.10 dS/m, and D. 2.85 dS/m obtained from Pandansimo and E. 3.25 dS/m obtained from Sleman, Yogyakarta, Indonesia, as comparation. Seedlings were transferred to the polybag after having four truly expanded leaves. Fruit development was observed every week, starting from the first day after flowering (DAF) to 35 DAF. Pericarpium and placenta thickness, fruit diameter, number, length, and width of the giant cell were recorded appropriately from the slides prepared using the paraffin method. Capsaicin content was determined at 14 and 35 DAF, performed with Gas Chromatography-Mass Spectrometry (GC-MS). The results show structural changes in the exocarpium; on the first day after flowering, there was only one layer of epidermis cells, but at 7 DAF, there was one layer of epidermis cells and one layer of collenchyma cells. Next, at 14 DAF, one layer of epidermis cells and two layers of collenchyma cells are observed. The structure of the mesocarpium, endocarpium, and placenta were not changed. The capsaicin content of the green fruit (14 DAF) was lower than the mature one (35 DAF) in all survival mediums. The highest capsaicin content at 14 and 35 DAF was obtained from a plant grown at medium C. Different growing mediums affected pericarpium and placenta thickness, number, length, and width of the giant cell fruit diameter.
Capsaicin - Inhibitory Factor for Somatic Embryogenesis in Pepper Anther Culture
Electronic Journal of Biology, 2013
The androgenesis of pepper (Capsicum annuum L.) is very limited process and it is followed by many inhibitory factors such as: pepper genotype; the structure and the stadium of microspores; the genetic predisposition for somatic embryogenesis; the hormonal regulation under in vitro condition; and growth conditions. Pungency in Capsicum fruits is due to the accumulation of the alkaloid capsaicin and its analogs. The biosynthesis of capsaicin is restricted to the genus Capsicum and results from the acylation of an aromatic moiety, vanillylamine, by a brached-chain fatty acid. The inhibitory influence of secondary metabolites, including capsaicin, is not explored enough, although in the literature there are data that some pepper genotypes as bell-shape and sweet ones have higher androgenic potential than the hot genotypes. The results from our research work, performed on nine pepper varieties which differ in pungency, showed that the androgenic potential of pepper anthers culture is dependent on the capsaicin content in the pepper fruits. Most probably the genetic predisposition for synthesis of capsaicin as secondary metabolite beside all the other factors is also inhibitory trait of the somatic embryogenesis. Our research showed that there is negative correlation between the capsaicin content of fruits of all the varieties under investigation and the percentage of embryogenic anthers and the number of formed embryos per 100 anthers. The hottest cultivar Feferona showed neither androgenesis nor callus formation, as compared to the other two hot cultivars where the callus formation is the main process. The callus formation of the sweet cultivars is moderate, while the androgenic response is poor to fair. The sweetest cultivar Féherözön showed excellent androgenic response with 31.09% androgenic and only 3.92% callused anthers. The aforementioned facts gives an idea that the capsaicin has an inhibitory effect in in vitro conditions and the more pungent cultivars possess less androgenic potential as compared to the sweet and bell-shaped ones.