Isolation of Protoplasts from Tissues of Mexican Prickly Poppy (Argemone mexicana L.): An Alkaloid-Producing Medicinal Plant.

Protoplasts are plant cells from which the pectocellulosic cell wall has been removed, thus keeping the plasma membrane intact. For plant secondary metabolites research, this system is a powerful tool to study the metabolites' dynamics inside the cells, such as the subcellular localization of proteins, characterization of gene function, transcription factors involved in metabolite pathways, protein transport machinery, and to perform single-cell omics studies. Due to its lack of a cell wall, better images of the interior of the cell can be obtained compared to the whole tissue.

A Novel Method for In Situ RT-PCR Based on Capsules from Centrifuge Tubes, Ideal for Transcripts Detection in Plant Tissues.

In situ RT-PCR presents advantages over other expression analysis methods due to its rapid processing and low-cost equipment. However, this technique is not without its challenges. A protocol based on a capsule made from centrifuge tubes that offers advantages over slides is presented.

An Update of the Sanguinarine and Benzophenanthridine Alkaloids' Biosynthesis and Their Applications.

Benzophenanthridines belong to the benzylisoquinolic alkaloids, representing one of the main groups of this class. These alkaloids include over 120 different compounds, mostly in plants from the Fumariaceae, Papaveraceae, and Rutaceae families, which confer chemical protection against pathogens and herbivores. Industrial uses of BZD include the production of environmentally friendly agrochemicals and livestock food supplements.

AmABCB1, an alkaloid transporter from seeds of Argemone mexicana L (Papaveraceae).

An ABCB-type transporter for sanguinarine, a benzophenanthridine alkaloid, was isolated from Argemone mexicana seeds. An ABCB-type transporter, AmABCB1, was identified in a transcriptome from unfolding seedlings of A. mexicana by its amino acid sequence identity to previously characterized alkaloid transporters from Coptis japonica and Thalictrum minus.

Alkaloid Biosynthesis in the Early Stages of the Germination of L. (Papaveraceae).

The synthesis of the benzylisoquinoline alkaloids, sanguinarine and berberine, was monitored in L. (Papaveracea) throughout the early stages of its hypocotyl and seedling development. Sanguinarine was detected in the cotyledons right after hypocotyl emergence, and it increased continuously until the apical hook unbent, prior to the cotyledonary leaves unfolding, when it abruptly fell.

Isolation and functional characterization of two dioxygenases putatively involved in bixin biosynthesis in annatto ( L.).

Carotenoid cleavage dioxygenases (CCDs) are enzymes that have been implicated in the biosynthesis of a wide diversity of secondary metabolites with important economic value, including bixin. Bixin is the second most used pigment in the world's food industry worldwide, and its main source is the aril of achiote ( L.) seeds.

Caffeine Extraction, Enzymatic Activity and Gene Expression of Caffeine Synthase from Plant Cell Suspensions.

Caffeine (1,3,7-trimethylxanthine) is a purine alkaloid present in popular drinks such as coffee and tea. This secondary metabolite is regarded as a chemical defense because it has antimicrobial activity and is considered a natural insecticide. Caffeine can also produce negative allelopathic effects that prevent the growth of surrounding plants.

Induction of Specialized Metabolism in In Vitro Cultures of Capsicum chinense Jacq.

A protocol for the elicitation of capsaicinoids, the pungent principle of peppers, as well as for the biosynthetic intermediaries vanillin and ferulic acid was developed for in vitro cell suspension cultures, and immobilized placentas of Capsicum chinense Jacq. in vitro cultures were exposed to different doses of methyl jasmonate and salicylic acid, which were effective in eliciting specialized metabolism in both of these cultures, resulting in an increased accumulation of the analyzed metabolites.

Relationship between aluminum stress and caffeine biosynthesis in suspension cells of Coffea arabica L.

Toxicity by aluminum is a growth-limiting factor in plants cultivated in acidic soils. This metal also promotes signal transduction pathways leading to the biosynthesis of defense compounds, including secondary metabolites. In this study, we observed that Coffea arabica L.

Development of newly sanguinarine biosynthetic capacity in in vitro rootless shoots of Argemone mexicana L. Mexican prickly poppy.

Objective: To analyze berberine and sanguinarine biosynthetic capacities of both in vitro shoot and root cultures of Argemone mexicana and tissues from entire plants at different developmental stages.

Results: Berberine and sanguinarine were equally distributed in roots and aerial tissues of developing plantlet whereas, in juvenile plants, sanguinarine was only detected in roots. This alkaloid distribution was consistent with that of biosynthetic transcripts in juvenile plants.

Capsaicin Synthesis Requires in Situ Phenylalanine and Valine Formation in in Vitro Maintained Placentas from Capsicum chinense.

Capsaicinoids (CAP) are nitrogenous metabolites formed from valine (Val) and phenylalanine (Phe) in the placentas of hot Capsicum genotypes. Placentas of Habanero peppers can incorporate inorganic nitrogen into amino acids and have the ability to secure the availability of the required amino acids for CAP biosynthesis. In order to determine the participation of the placental tissue as a supplier of these amino acids, the effects of blocking the synthesis of Val and Phe by using specific enzyme inhibitors were analyzed.

Yeast extract induction of sanguinarine biosynthesis is partially dependent on the octadecanoic acid pathway in cell cultures of Argemone mexicana L., the Mexican poppy.

Objective: To analyze the involvement of the octadecanoic (OCDA) pathway in the accumulation of sanguinarine induced by yeast extract (YE) in cell suspension cultures of Argemone mexicana (Papaveraceae).

Results: Exposure to YE promoted sanguinarine accumulation. This was not observed when they were exposed to methyl jasmonate (MeJa).

Salicylic-acid elicited phospholipase D responses in Capsicum chinense cell cultures.

The plant response to different stress types can occur through stimulus recognition and the subsequent signal transduction through second messengers that send information to the regulation of metabolism and the expression of defense genes. The phospholipidic signaling pathway forms part of the plant response to several phytoregulators, such as salicylic acid (SA), which has been widely used to stimulate secondary metabolite production in cell cultures. In this work, we studied the effects of SA treatment on [(32)-P]Pi phospholipid turnover and phospholipase D (PLD) activity using cultured Capsicum chinense cells.

Nitrate promotes capsaicin accumulation in Capsicum chinense immobilized placentas.

In chili pepper's pods, placental tissue is responsible for the synthesis of capsaicinoids (CAPs), the compounds behind their typical hot flavor or pungency, which are synthesized from phenylalanine and branched amino acids. Placental tissue sections from Habanero peppers (Capsicum chinense Jacq.) were immobilized in a calcium alginate matrix and cultured in vitro, either continuously for 28 days or during two 14-day subculture periods.

Antioxidant capacity and total phenolic content in fruit tissues from accessions of Capsicum chinense Jacq. (Habanero pepper) at different stages of ripening.

In the past few years, there has been a renewed interest in studying a wide variety of food products that show beneficial effects on human health. Capsicum is an important agricultural crop, not only because its economic importance, but also for the nutritional values of its pods, mainly due to the fact that they are an excellent source of antioxidant compounds, and also of specific constituents such as the pungent capsaicinoids localized in the placental tissue. This current study was designed to evaluate the antioxidant capacity and total phenolic contents from fruits tissues of two Capsicum chinense accessions, namely, Chak k'an-iik (orange) and MR8H (red), at contrasting maturation stages.

Salicylic acid induces vanillin synthesis through the phospholipid signaling pathway in Capsicum chinense cell cultures.

Signal transduction via phospholipids is mediated by phospholipases such as phospholipase C (PLC) and D (PLD), which catalyze hydrolysis of plasma membrane structural phospholipids. Phospholipid signaling is also involved in plant responses to phytohormones such as salicylic acid (SA). The relationships between phospholipid signaling, SA, and secondary metabolism are not fully understood.

Pharmaceutical applications of the benzylisoquinoline alkaloids from Argemone mexicana L.

Argemone mexicana L. (Papaveraceae), accumulates benzylisoquinoline alkaloids, (BIA) derived from tyrosine. Although it was originated in the western region of the USA-Mexico border, it has spread to tropical and subtropical areas around the world.

Enrichment of carbon dioxide in the atmosphere increases the capsaicinoids content in Habanero peppers (Capsicum chinense Jacq.).

Background: The effects of the increase of atmospheric CO2 on agricultural productivity have been mainly analyzed through its impact on biomass yield, and little attention has been directed to quality traits, such as nutritional or organoleptic attributes. For this study, plants of hot Habanero pepper (Capsicum chinense Jacq.) were grown in growth chambers under three different CO2 levels: 380 (normal atmospheric value), 760 and 1140 µmol mol(-1), and their effects on pod yield, size, color and pungency, were monitored.

Establishment of a sanguinarine-producing cell suspension culture of Argemone mexicana L (Papaveraceae): induction of alkaloid accumulation.

A protocol for the induction of a cell suspension culture of Argemone mexicana is described. This suspension has been kept for over 3 years producing sanguinarine, a benzophenanthridine-type alkaloid. Sanguinarine levels can be increased by exposing these cultures to yeast or fungal elicitation.

Growth measurements: estimation of cell division and cell expansion.

The main parameters for the estimation of growth within in vitro cultures are reviewed. Procedures to measure these parameters are described, emphasizing in each case their convenience of use, depending on the features of the culture evaluated.

The sequential exposure to jasmonate, salicylic acid and yeast extract promotes sanguinarine accumulation in Argemone mexicana cell cultures.

The effects of the sequential application of methyl jasmonate (MeJa), salicylic acid (SA) and yeast extract (YE) to Argemone mexicana cell cultures were compared to either the sole application of each elicitor, or to the three-partite mixture. The highest sanguinarine accumulation occurred using the sequential treatment (ninefold over unexposed control cultures), followed by the single application of YE (fivefold). The elicitor mixture produced less sanguinarine than sole exposure to YE but higher than MeJa alone.

Phospholipidic signaling and vanillin production in response to salicylic acid and methyl jasmonate in Capsicum chinense J. cells.

The phospholipidic signal transduction system involves generation of second messengers by hydrolysis or changes in phosphorylation state. Several studies have shown that the signaling pathway forms part of plant response to phytoregulators such as salicylic acid (SA) and methyl jasmonate (MJ), which have been widely used to stimulate secondary metabolite production in cell cultures. An evaluation was made of the effect of SA and MJ on phospholipidic signaling and capsaicinoid production in Capsicum chinense Jacq.

Capsaicin accumulation is related to nitrate content in placentas of habanero peppers (Capsicum chinense Jacq.).

Background: The presence of capsaicin, the pungent principle of peppers, is restricted to the fruits of hot cultivars. This compound, which is produced in the fruits' placenta, requires 3 mol of nitrogen to be formed. Hence nitrogen availability may affect pepper pungency through its content in the fruit tissues.

Fusarium oxysporum homogenates and jasmonate induce limited sanguinarine accumulation in Argemone mexicana cell cultures.

In vitro cultures of Argemone mexicana (Papaveraceae) were induced from leaves of mature plants. Sanguinarine, a benzophenanthridine, was the main alkaloid in the cultures, even in the absence of inducers of secondary metabolism. The accumulation of this metabolite was increased by adding methyl jasmonate and fungal elicitors, although in a limited fashion in comparison to other sanguinarine-producing species.