Biosynthesis of Betalains Elicited by Methyl Jasmonate in Two Species of Alternanthera Genus: Antagonistic Regulations Result in Increase of Pigments.

Natural pigments are components very important in the dye industry. The betalains are pigments found in plants from Caryophyllales order and are relevant in the food manufacturing. The main source of betalains is beetroot, which has unfavorable aftertaste. Therefore, the demand for alternative species producing betalains has increased. Elicitor molecules such as methyl jasmonate (MeJA) induce metabolic reprogramming acting in the biosynthesis of specialized metabolites and can enhance pigment concentrations. Here, we used this strategy to identify if treatment with MeJA at 100 µM can promote the accumulation of betalains and other bioactive compounds in Alternanthera philoxeroides and Alternanthera sessilis. We performed the gene expression, concentration of betalains, phenols, flavonoids, amino acids (phenylalanine and tyrosine), and antioxidant activity. The results showed that MeJA treatment increased betalains and other bioactive compounds in the two Alternanthera species but A. sessilis had a better performance. One key factor in this pathway is related to the phenylalanine and tyrosine concentration. However, the species have distinct metabolic regulation: in A. philoxeroides, high concentrations of betalain pigments increase the tyrosine concentration and gene expression (include ADH) under MeJA and in A. sessilis, high concentrations of betalain pigments reduce the gene expression and tyrosine concentration after 2 days under MeJA. This study brings new questions about betalain biosynthesis and sheds light on the evolution of this pathway in Caryophyllales.

Validation of reference genes for RT-qPCR studies in Stevia rebaudiana in response to elicitor agents.

Stevia rebaudiana is an important source of natural steviol glycosides and is of increasing interest in various fields of study. Therefore, understanding the molecular processes regulating its metabolism is of great importance. In this study, the stability of seven reference genes (18S ribosomal RNA, Actin, Aquaporin, Calmodulin, Eukaryote elongation factor 1-α, Malate dehydrogenase, and Ubiquitin) under the effect of three stress-related elicitors (methyl jasmonate, salicylic acid, and spermidine) was evaluated in stevia plants. We used RefFinder software, which makes use of the four main currently available algorithms for reference gene selection: geNorm, NormFinder, BestKeeper, and the Comparative ∆Ct method. The results indicated that Ubiquitin and Actin can be used as reference genes under all tested experimental conditions. The genes, 18S ribosomal RNA, traditionally used as a reference gene, along with Calmodulin, showed the lowest stability. The expression of Deoxyxylulose-5-phosphate synthase and Kaurenoic acid hydroxylase genes was used to confirm the validated reference genes, showing that inadequacy of the reference gene may lead to erroneous results. This is the first study on the stability of reference genes in Stevia rebaudiana plants, and is of great relevance for further analysis of the gene expression of the steviol glycoside biosynthetic pathway.

Production of pigments in Alternanthera sessilis calli mediated by plant growth regulators and light / Produção de pigmentos em calos de Alternanthera sessilis mediados por reguladores de crescimento e luz

Among the compounds produced by plants, pigments such as betalains have received attention from both food and pharmaceuticals industries. The Alternanthera sessilis species produces these pigments, though in small quantities, and so it is necessary to increase production. Thus, many studies use elicitors that are capable of triggering physiological or morphological responses in plants. The objective was to establish callus production in A. sessilis grown under different combinations of growth regulators and light qualities and to assess whether these factors can increase betalain and flavonoid production. Leaf and internodal explants in MS medium with different growth regulators were used to obtain calli, which were subsequently transferred to a betacyanin induction medium remaining for 40 days under different light qualities (white, blue, red, and dark). The most suitable treatment for callus formation and subsequent betalain and flavonoid induction was to combine a medium containing 6.7 µmol L-1 2,4-D and 9.0 µmol L-1 BAP and blue light. Physical elicitation by light combined with appropriate concentration of growth regulators on calli can increase production of commercially important metabolites.

Dentre os compostos produzidos pelas plantas, os pigmentos, como as betalaínas, vêm recebendo destaque tanto pela indústria alimentícia como farmacêutica. A espécie Alternanthera sessilis produz esses pigmentos, porém em pequenas quantidades, sendo necessário incrementar a produção. Para isso, muitos estudos utilizam elicitores que são capazes de desencadear respostas fisiológicas ou morfológicas nas plantas. O objetivo do trabalho foi estabelecer a produção de calos de A. sessilis crescidos quando submetidos a diferentes combinações de reguladores de crescimento e qualidades de luz, e avaliar se esses fatores são capazes de incrementar a produção de betalaínas e flavonoides. Foram utilizados explantes foliares e internodais em meio MS com diferentes reguladores de crescimento para obtenção dos calos que, posteriormente, foram transferidos para meio de indução de betacianina, onde permaneceram por 30 dias sob diferentes qualidades de luz (branca, azul, vermelha e escuro). O tratamento mais propício para formação de calos e consequente indução de betalaínas e flavonoides foi a combinação do meio contendo 6,7 µmol L-1 2,4-D e 9,0 µmol L-1 BAP e a luz azul. Conclui-se que a elicitação física pela luz em conjunto com a concentração adequada de reguladores de crescimento em calos é capaz de incrementar a produção de metabólitos de interesse comercial.