A rapid double-staining technique to improve seed viability testing in terrestrial orchids

Magrini, Sara; Barreca, D.; Zucconi, Laura

doi:10.1080/11263504.2019.1587541

Cited by 17 publications

(10 citation statements)

References 29 publications

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“…Numerous evidence from various orchid species suggest that the permeability of testa can play a role in some difficult-to-germinate species (e.g., Rasmussen 1995 ; Lee et al 2007 ; Barsberg et al 2013 ; Magrini et al 2019 ). This is even more likely in Vanilla which possess hard seeds adapted to endozoochory (Pansarin 2021 ; Yeh et al 2021 ).…”

Section: Discussionmentioning

confidence: 99%

“…Ballard 1987 ; Coke 1990 ; Rasmussen 1992 ; Ponert et al 2013 ; Malmgren and Nyström 2020 ), or incubation at specific temperature (Nakamura 1982; Johnson and Kane 2012 ). The positive effect of bleaching in hypochlorite solutions, soaking in ethanol and weak acid could be attributed mainly to a degradation of the seed coat (e.g., Lee et al 2007 ; Barsberg et al 2013 ; Magrini et al 2019 ; Pierce et al 2019 ). The hypochlorite solutions exhibit a high pH and strong oxidative effects on a wide range of compounds including polymers of plant cell walls (Arditti 1967 ; Rasmussen 1995 ; Zeng et al 2014 ).…”

Section: Backgoundmentioning

confidence: 99%

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Acid scarification as a potent treatment for an in vitro germination of mature endozoochorous Vanilla planifolia seeds

2023

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Background Vanilla planifolia is the most widely cultivated species of vanilla with high economic importance. However, seed germination under artificial conditions is difficult and yields low germination percentages. The seeds are adapted to endozoochorous dispersal, and we therefore tried to simulate the conditions in the digestive tract by acid scarification of seeds. Results Immature seeds lacking dormancy, used as a control, showed the highest germination percentage. Among the treatments tested for mature seeds, the hydrochloric acid treatments were significantly the best in breaking dormancy and inducing germination, irrespective of the acid concentration and the presence of pepsin. Conventional treatment with a hypochlorite solution induced much lower germination percentage. Sulphuric acid at concentration 50% was too strong and caused damage to the seeds. Important factor is also high cultivation temperature 30 °C as there was nearly no germination at 25 °C. Conclusions Our protocol significantly improves the efficiency of generative propagation of vanilla and allows for significantly higher germination percentages than previously described. The strongly positive effect of hydrochloric acid may be related to the adaptation of seeds to endozoochorous dispersal.

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Section: Discussionmentioning

confidence: 99%

Section: Backgoundmentioning

confidence: 99%

Acid scarification as a potent treatment for an in vitro germination of mature endozoochorous Vanilla planifolia seeds

2023

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“…Trypan blue exclusion test was used to evaluate the viability of tomato mesophyll cells as described by Magrini et al (2019).…”

Section: Cell Viability Test Trypan Bluementioning

confidence: 99%

Antifungal activity of dimethyl sulfoxide againstBotrytis cinereaand phytotoxicity on tomato and lettuce plants

Petruccelli

Brasili

Varone

et al. 2020

Plant Biosystems - An International Journal Dealing with all As

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For the first time the antifungal activity of dimethyl sulfoxide (DMSO) was evaluated against Botrytis cinerea, that it is one of the phytopathogenic fungi which causes the greatest damage in agriculture. In in-vitro tests, the greatest inhibitory effect of DMSO on fungal grow was recorded at pH 6. A significant growth inhibition was caused by 0.5% DMSO at 96 h post-inoculation. With higher DMSO concentrations, significant effects were recorded starting from 48 h post-inoculation. As the medium pH decreased, the inhibitory effect of DMSO also decreased. At pH 4 and 5 significant growth inhibition was caused by 1% DMSO starting from 72 h post-inoculation. At all tested pH values, a total growth inhibition was caused by ! 2% DMSO. On tomato leaves infected with B. cinerea, 2% DMSO significantly decreased the extent of damaged leaf area. The administration of DMSO at concentrations ranging from 0.5 to 2% through nebulization on leaves of young Solanum lycopersicum and Lactuca sativa plants did not change the chlorophyll fluorescence (Fv/Fm and U PSII ) at any of the experimental times. Overall, the data obtained suggest that, at the concentrations tested, DMSO is toxic to B. cinerea, while it is well tolerated by lettuce and tomato plants.

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“…Following the double staining approach proposed by Magrini et al (2019a), scarified seeds were soaked in a solution of 1% TTC red chloride (2,3,5-triphenyltetrazolium chloride, pH 7) and kept in the dark at 30°C for 24 h. After one day the solution was removed and the seeds were washed three times in distilled water for 3 min. each, followed by centrifugation at 6000 rpm for 1 min.…”

Section: Seed Viabilitymentioning

confidence: 99%

Less is more: low-cost in vitro propagation of an Endangered Italian orchid

Calevo¹,

Bazzicalupo²

2020

Nat. Conserv. Res.

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Orchids are among the most widely distributed plants but also the most threatened by biotic and abiotic factors. Their interactions with specific symbiotic fungi during germination in natural environments make their propagation challenging for conservation biologists. In this study, in the framework of the European conservation project LIFEorchids, the germination protocol for the endangered Mediterranean species Orchis patens was optimised, and the effect of controlled constant temperature in growth chambers with fluctuations of temperature occurring in natural environments was compared. Seed viability was also evaluated by using a double-staining technique. Minimum, maximum and average daily temperatures were recorded for six months, from July to December, and germination percentages, as well as developmental stages of seeds, were annotated, for both conditions, every month. Even if germination was obtained under both conditions, the effect of temperature fluctuations was evident, by increasing the germination percentage of seeds from about 10% under controlled conditions to 59%. The latter result was almost comparable with seed viability data, indicating a strong impact of temperature fluctuations in breaking seed dormancy. Our results are a confirmation that temperature oscillations play a more important role than the average temperature in seed germination and suggest avoiding a constant temperature to optimise germination protocols for European orchid species.

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A rapid double-staining technique to improve seed viability testing in terrestrial orchids

Cited by 17 publications

References 29 publications

Acid scarification as a potent treatment for an in vitro germination of mature endozoochorous Vanilla planifolia seeds

Acid scarification as a potent treatment for an in vitro germination of mature endozoochorous Vanilla planifolia seeds

Antifungal activity of dimethyl sulfoxide againstBotrytis cinereaand phytotoxicity on tomato and lettuce plants

Less is more: low-cost in vitro propagation of an Endangered Italian orchid

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