A rapid in vitro protocol for propagation of Piper aduncum and Piper hispidinervum, two species from Amazon region with multipurpose uses

Silva, Tatiane Loureiro da; Balzon, Talita Aparecida; Scherwinski‐Pereira, Jonny Everson

doi:10.5897/ajb12.1888

Cited by 4 publications

(2 citation statements)

References 15 publications

(15 reference statements)

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“…However, in order to ensure the selection of superior genotypes and their production on a commercial scale, it is necessary to develop efficient systems of vegetative propagation of the species, so as to maintain the genetic characteristics of interest, such as high levels of dillapiole, since it is not known with certainty whether self-pollination or cross-pollination predominates in the species (Semple 1974;Figueiredo and Sazima 2000;Wadt et al 2004). Nevertheless, studies on the vegetative propagation of P. aduncum are scarce, limited to cuttings with low rooting percentages (Gomes and Krinski 2018) and a few micropropagation protocols for proliferation from nodal segments (Ferreira et al 2006;Silva et al 2012).…”

Section: Introductionmentioning

confidence: 99%

Somatic embryogenesis and plant regeneration in Piper aduncum L

Sousa

Meira

et al. 2020

In Vitro Cell.Dev.Biol.-Plant

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An efficient protocol is reported for in vitro plant regeneration through somatic embryogenesis in Piper aduncum, a Brazilian Amazon species with high economic potential. The species is important due to a variety of components found in its essential oil, with emphasis on dillapiole. Leaf explants from five accessions identified for high oil yield and levels of dillapiole were evaluated for their embryogenic potential. To induce embryogenic calli, the explants were cultivated in MS medium supplemented with 5 mg L −1 of 1-naphthaleneacetic acid (NAA) and 2.5 mg L −1 of N6-benzylaminopurine (BAP) for 80 d. For somatic embryogenesis, the embryogenic calli were transferred to MS medium with 10 mg L −1 of NAA and 2.5 mg L 1 of BAP and incubated for 45 d. The obtained somatic embryos were germinated in MS medium without regulators by 45 d and the obtained plantlets were subjected to acclimatization. Somatic embryos and calli from this process were subjected to anatomical and histochemical analyses. Biochemical analyses (total soluble sugars, starch, total amino acids, and proteins) were also performed to identify markers for embryogenic competence acquisition. In addition, the germination of somatic embryos was evaluated in a semi-solid and liquid system (R.I.T.A.® temporary immersion bioreactors). The obtained plants were evaluated for genetic fidelity using ISSR markers. The present study indicate that the accessions did not differ in embryogenic potential, with a mean percentage of calli with somatic embryos of 82.4%. Anatomical analyses confirmed the occurrence of the embryogenic route and the histochemical analyses identified starch grains in somatic embryos at different developmental stages. The biochemical analyses showed high total soluble sugars and total amino acids in embryogenic calli, marks of the acquisition of the embryogenic competence of P. aducum. The R.I.T.A.® temporary immersion bioreactors were highly efficient in the regeneration of somatic plants, with 100% germination. The plants regenerated in the semi-solid and liquid systems showed high genetic homogeneity. The survival rate of the acclimatized plants was 100%.

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Section: Introductionmentioning

confidence: 99%

Somatic embryogenesis and plant regeneration in Piper aduncum L

Sousa

Meira

et al. 2020

In Vitro Cell.Dev.Biol.-Plant

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“…12 O efeito fisiológico depende da concentração de cada regulador no meio, sendo que cada parte da planta tem uma resposta diferente às alterações das concentrações de auxinas e citocininas. 31 Silva Balzon e Scherwinski-Pereira, 30 estudando a propagação in vitro de P.…”

Section: Enraizamento In Vitrounclassified

Cultivo in vitro de Piper aduncum espécie com potencial econômico da Amazônia Sul-Ocidental

Raposo

Yamura²,

Silva³

et al. 2019

evid.

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A pimenta de macaco (Piper aduncum) é uma espécie nativa da Amazônia, que possui grande potencial econômico em razão de o seu óleo essencial conter substâncias bioquímicas com propriedades fungicida e inseticida. Seu cultivo é sustentável, podendo ser explorado de forma não destrutiva e renovável. Com a crescente demanda para a produção de óleo essencial dessa espécie, existe a necessidade da implantação de cultivos comerciais, e com isso se tem como limitação a oferta de mudas em quantidade e com qualidade. A produção de mudas em laboratório por meio da micropropagação supre tal necessidade, já que permite a obtenção de material isento de doenças e em larga escala. O objetivo deste trabalho foi verificar os parâmetros necessários para o cultivo in vitro de sementes dessa espécie. Foram realizados três experimentos: germinação e crescimento inicial, multiplicação e enraizamento in vitro. Foram utilizados os meios de cultura MS e WPM em diferentes concentrações salinas. Para multiplicação in vitro foram testadas quatro citocininas em cinco diferentes concentrações, e para o enraizamento in vitro foram utilizados três tipos de auxinas em seis diferentes concentrações. Todas as variáveis foram submetidas à análise de variância e, quando o valor de F foi significativo, utilizou-se a comparação das médias pelo teste de Tukey ao nível de 5% de probabilidade de erro, para tratamentos qualitativos. Já para os tratamentos quantitativos, foi realizada análise de regressão polinomial ao nível de 5% de probabilidade de erro. Verificou-se que a germinação in vitro das sementes de P. aduncum deve ser realizada se utilizando sais do meio MS; para a multiplicação e o enraizamento in vitro não é necessária a adição de reguladores de crescimento nesse meio, provavelmente os níveis endógenos desses reguladores nos explantes são suficientes para suprir suas necessidades de crescimento e desenvolvimento. Observou-se também que a utilização de citocininas propicia o aparecimento de calos na base dos explantes.

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Indirect somatic embryogenesis of Piper hispidinervum L. and evaluation of the regenerated plants by flow cytometry

Sousa

Nogueira

Silva‐Cardoso

et al. 2022

Journal of Genetic Engineering and Biotechnology

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Background Piper hispidinervum is a species native from the Amazon region with great economic potential, given its scientifically proven insecticidal properties. In this study, an efficient protocol of plant regeneration via indirect somatic embryogenesis has been established for the first time. In a first experiment, for the induction of calluses, foliar explants of non-discriminated accesses of P. hispidinervum were inoculated in MS medium supplemented with α-naphtalenacetic acid (NAA) and 6-benzylaminopurine (BAP), in different combinations. For a second experiment, foliar explants from five different accesses of P. hispidinervum (PH17, PH21, PH28, PH37, and PH39) were analyzed regarding the formation of calluses when cultivated in MS medium with 5 mg L−1 NAA + 2.5 mg L−1 BAP. To obtain somatic embryos-like structures, calluses were cultivated in MS medium with 10 mg L−1 NAA + 2.5 mg L−1 of BAP. The somatic embryos-like structures obtained were inoculated in MS medium devoid of growth regulators and the plantlets were subjected to acclimatization. Calluses and somatic embryos-like structures were subjected to anatomical analysis and genetic stability of regenerated plants was analyzed by flow cytometry. Results The treatments 2.5 mg L−1 BAP and 5 mg L−1 NAA + 2.5 mg L−1 BAP, after 60 days of cultivation, provided each 32% of primary callus, not being verified the formation of calluses in medium devoid of BAP. It was found that accesses differed among them with respect to the formation of primary calluses, with emphasis on accesses PH28, PH37, and PH39, with mean percentage of 95.3%. Regarding the percentage of embryogenic calluses and formation of somatic embryos-like structures, there were no statistical differences between accesses, with mean values of 90.6% and 77.3%, respectively. The somatic embryos-like structures of P. hispidinervum have conspicuous morphoanatomical similarities with the zygotic embryo, and flow cytometry analysis showed no significant variation in nuclear DNA size among plants regenerated in vitro and plants coming from seed germination, which indicates ploidy level stability. Conclusion This protocol is the first cited in the literature that demonstrates an efficient micropropagation process by somatic embryogenesis of P. hispidinervum. It can be used either to enable large-scale vegetative production or to subsidize germplasm conservation or genetic engineering of P. hispidinervum.

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A rapid in vitro protocol for propagation of Piper aduncum and Piper hispidinervum, two species from Amazon region with multipurpose uses

Cited by 4 publications

References 15 publications

Somatic embryogenesis and plant regeneration in Piper aduncum L

Somatic embryogenesis and plant regeneration in Piper aduncum L

Cultivo in vitro de Piper aduncum espécie com potencial econômico da Amazônia Sul-Ocidental

Indirect somatic embryogenesis of Piper hispidinervum L. and evaluation of the regenerated plants by flow cytometry

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