In vitro elimination of Piper yellow mottle virus from infected black pepper through somatic embryogenesis and meristem-tip culture

Sasi, Shina; Bhat, Alangar Ishwara

doi:10.1016/j.cropro.2017.09.004

Cited by 19 publications

(16 citation statements)

References 15 publications

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“…Somatic embryos develop from somatic cells through embryogenic stages similar to zygotic embryogenesis (Von Arnold et al 2002) and occur without the fusion of gametes (Williams and Maheswaran 1986). In addition to the mass propagation of genotypes of economic interest, which reduce the excessive use of natural populations, this technique has allowed the generation of new technologies, such as genetic transformation, synthetic seed production (Aguilar-Hernández and Loyola-Vargas 2018), and viral cleaning (Sasi and Bhat 2018). Furthermore, it presents the additional advantage of automation during the production system through the use of bioreactors.…”

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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“…Indexing via PCR has been effective in detecting the PYMoV virus in major black pepper producing countries, especially when compared to serological tests such as ELISA (Enzyme-Linked Immunosorbent Assay) (Silva et al, 2002;Sasi et. Al., 2018).…”

Section: Resultsmentioning

confidence: 99%

“…The health of seedlings must be guaranteed during the entire propagation process in order to prevent the spread of the virus that reflects in reduced production. In the case of black pepper, the main means of crop propagation is vegetative, which can become a route of PYMoV transmission and responsible for the introduction of the pathogen in new planting areas if mother plants are infected; therefore, this culture demands virus-free mother plants (Bhat et al, 2018;Sasi et al, 2018).…”

Section: Introductionmentioning

confidence: 99%

IDENTIFICATION AND MICROPROPAGATION OF VIRUS-FREE BLACK PEPPER GENOTYPES (Piper nigrum L.)

et al. 2020

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Virus infections are a major constraint for black pepper cultivation (Piper nigrum L.), a vegetatively propagated crop; therefore, the in vitro selection of virus- free plants is an alternative to reestablish phytosanitary quality of pepper culture. The aim of this study was to select and micropropagate PYMoV virus-free pepper plants. ‘Apra’, ‘Bragantina’, ‘Cingapura’, ‘Clonada’, hybrids and ‘Kottanadan’ genotypes were used in the indexing process. From these, ‘Cingapura’, ‘Clonada’, hybrids and ‘Kottanadan’ were virus free, and only ‘Cingapura’ and ‘Clonada’ were selected for micropropagation. In the multiplication phase, ‘Cingapura’ genotype showed higher multiplication rate in relation to ‘Clonada’ in in vitro rooting and in acclimatization, response was similar in both genotypes. It was possible to identify virus-free genotypes of commercial interest, selecting them for micropropagation, and in this stage, influence of genotype is of great importance.

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“…The fungi in the culture medium compete with explants for space, water, light, and nutrients, causing the death of explants (Cassells 2012; Tomasi et al 2017). One option to eradicate these microorganisms is through the culture of meristems, thermotherapy, and use of antifungal agents (Cassells 2012; Smith 2013: Sasi and Bhat 2018). The culture of meristems has been used primarily to eradicate viruses (Sasi and Bhat, 2018); for its part, thermotherapy occasionally damages the explants (Hu et al 2019; Kaur et al 2019).…”

Section: Discussionmentioning

confidence: 99%

“…One option to eradicate these microorganisms is through the culture of meristems, thermotherapy, and use of antifungal agents (Cassells 2012; Smith 2013: Sasi and Bhat 2018). The culture of meristems has been used primarily to eradicate viruses (Sasi and Bhat, 2018); for its part, thermotherapy occasionally damages the explants (Hu et al 2019; Kaur et al 2019). As regards the use of antifungal agents, these can lead to the development of resistance in some fungal strains (Caniça et al 2019; Chechi et al 2019).…”

Section: Discussionmentioning

confidence: 99%

Antifungal activity of silver nanoparticles during in-vitro culture of Stevia rebaudiana Bertoni

Sánchez-Segura

et al. 2019

Preprint

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30Contamination by fungi and bacteria during the in-vitro propagation of plants leads to considerable losses of biological 31 material and precludes phytosanitary certification. The anti-microbial effect of silver nanoparticles (AgNPs) may be 32 an alternative for the eradication of in-vitro contaminants. This study evaluated the microbicidal activity of AgNPs on 33 a recurrent fungus during the micropropagation of stevia (Stevia rebaudiana Bertoni). First, the fungus was isolated 34 and identified at a molecular level by the sequencing and analysis of the ITS4/ITS5 rDNA region. The results of the 35 phylogenetic analysis of various fungi species showed that the strain under study (16-166-H) belongs to the genus 36 Sordaria and is 86.74% similar to S. tomento-alba (strain CBS 260.78). Subsequently, the inhibition of the growth of 37 S. tomento-alba was tested under different concentrations of AgNPs (0, 25, 50, 100, and 200 mg L -1 ), observing that 38 50 and 100 mg L -1 achieve ca. 50% growth inhibition (IC 50 ), while 200 mg L -1 produces a drastic inhibition. On the 39 other hand, the shape and size of AgNPs was examined using transmission electron microscopy (TEM), and the 40 transport and accumulation of AgNPs in S. tomento-alba cells were monitored through multiphoton microscopy. The 41 morphological and fluorescence analyses showed that AgNPs display different sizes, with larger nanoparticles retained 42 in fungal cell walls while smaller AgNPs penetrate into fungal cells. Probably, apoplastic and symplastic mechanisms 43 involved in the accumulation and transport of AgNPs affect the metabolic processes of the fungus, thus inhibiting its 44 growth. These results suggest that AgNPs possess antifungal activity and can be used in the eradication of contaminants 45 during the in-vitro culture of plant species.46 47

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In vitro elimination of Piper yellow mottle virus from infected black pepper through somatic embryogenesis and meristem-tip culture

Cited by 19 publications

References 15 publications

Somatic embryogenesis and plant regeneration in Piper aduncum L

Somatic embryogenesis and plant regeneration in Piper aduncum L

IDENTIFICATION AND MICROPROPAGATION OF VIRUS-FREE BLACK PEPPER GENOTYPES (Piper nigrum L.)

Antifungal activity of silver nanoparticles during in-vitro culture of Stevia rebaudiana Bertoni

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