Induction and growth pattern of callus from Piper permucronatum leaves

Abstract

ABSTRACT:Piper permucronatum is a perennial shrub, a medicinal plant native to the Amazon Rainforest. Traditionally, the tea of its leaves is used to combat menstrual and intestinal cramps, stomach pain, digestive problems, diarrhea, hemorrhage, and nausea. Its leaf's essential oil is effective against Aedes aegypti larvae; its flavones and flavanones have a fungicidal effect against Clamidosporium cladosporioides and C. sphaerospermum; its hexanic extract is effective against Leishmania amazonensis. The objective of this study was to provide a protocol for callus induction from P. permucronatum leaves and an identification of the callus growth pattern, focusing on the deceleration phase, when the callus cells must be subcultured into liquid medium in order to produce a cell suspension cultures. Leaf explants were inoculated in a solid MS medium supplemented with factorial combinations of 2,4-D, BA, NAA and GA 3 . Callus formation was evaluated weekly until the 49 th day. Subsequently, new explants were inoculated at the hormonal combination that resulted in the highest callus cell proliferation and, every seven days during a period of 70 days, samples were dried and weighed to determine the callus growth pattern. NAA and GA 3 were not effective for callus induction. Combinations of 2,4-D and BA resulted in callus induction and proliferation. The highest percentage of callus induction was observed with the combination of 4.52 µM 2,4-D and 4.44 µM BA. The calluses thereby produced were friable and whitish. The callus growth pattern followed a sigmoid shape. The deceleration phase started on the 56 th day of culture.Keywords: Piperaceae, growth regulators, callogenesis.RESUMO: Indução e padrão de crescimento de calos de folhas de Piper permucronatum. Indução e padrão de crescimento de calos de folhas de Piper permucronatum. Piper permucronatum é um arbusto perene, uma planta medicinal native da Floresta Amazônica. Tradicionalmente, o chá de suas folhas é usado em casos de cólicas menstruais e intestinais, dores de estômago, problemas digestivos, diarreia, hemorragia e náusea. O óleo essencial das folhas é efetivo contra a larva de Aedes aegypti; suas flavonas e flavanonas têm efeito fungicida contra Clamidosporium cladosporioides e C. sphaerospermum; seu extrato hexânico é efetivo contra Leishmania amazonensis. O objetivo deste trabalho foi determinar um protocolo para indução de calos em folhas de P. permucronatum e identificar o padrão de crescimento dos calos, com foco na fase de desaceleração, quando as células de calo devem ser subcultivadas em meio líquido para produzir culturas de células em suspensão. Explantes foliares foram inoculados em meio MS sólido suplementado com combinações fatoriais de 2,4-D, BAP, ANA e GA 3 . A formação de calos foi avaliada semanalmente até o 49º dia. Posteriormente, novos explantes foram inoculados na combinação hormonal que resultou na maior proliferação de células de calo e, a cada sete dias durante 70 dias, amostras foram secas e pesadas para determinar o padrão de crescime...

Establishment of a suitable regeneration protocol for rapid propagation of Piper nigrum L. through in vitro culture

“…Concentrations below and above 2.0 mg/l 2,4-D reduced callus formation. Various reports have shown that MS medium containing 2,4-D is most effective for induction of callus and development, such as Subasinghe et al (2004) and Hussain et al (2011) in the case of P. nigrum; Delgado-Paredes et al (2013) in the case of P. cernuum and P. aduncum; Santos et al (2015) in the case of P. permucronatum. However, in this study, during the direct shoot induction experiment, it was found that gelled MS medium conjugated with 1.0 mg/l BAP and 1.0 mg/l NAA induced some calli from the nodal segments (Table 2).…”

Effect of Abiotic and Biotic Elicitors on Callus and Suspension from Piper Betle L. Var. Nigra

“…The results of the cell suspension culture of Piper permucronatum and Piper solanum showed an increase in secondary metabolites, also in the callus of other plants (Santos et al, 2016); (Balbuena et al, 2009); (Jamil et al, 2018); (Khanpour-Ardestani et al, 2015). Callus extracts from cell suspension cultures of Nigella sativa L. were reported to have optimum antimicrobial and antioxidant activity (Chaudhry et al, 2015).…”

“…Piper plant consists of more than 1000 species scattered in various tropical regions, several species of this genus are used as food, medicine, stimulant, analgesic, anti-inflammatory, antiseptic, antidepressant, and antioxidant (Meena et al, 2010); (Santos et al, 2016). Various extracts of Piper have high antioxidant activity (Barua et al, 2014); (Akbar et al, 2014); (Hafizah et al, 2010); (Anggreni et al, 2019); (Dasgupta & De, 2004).…”