Effects of temperature, darkness and gelling agent on long-term storage of in vitro shoot cultures of Australian woody plant species

Penicillium expansum is the principal cause of blue mould rot and associated production of patulin, a weak mycotoxin, in apples worldwide. P. expansum growth and patulin production is observed during improper or long-term storage of apples. We have investigated the extent to which each successive step during long-term storage contributes to patulin production in various P. expansum isolates. Fungal isolates collected on apples from several Belgian orchards/industries were identified to species level. Random amplification of polymorphic DNA (RAPD) analysis and β-tubulin gene sequencing identified P. expansum and Penicillium solitum as the most prevalent Penicillium species associated with Belgian apples. All 27 P. expansum isolates and eight reference strains were characterised for their patulin production capacity on apple puree agar medium for five days under classical constant temperature and atmosphere conditions. Under these conditions, a large range of patulin production levels was observed. Based on this phenotypic diversity, five P. expansum isolates and one reference strain were selected for in vitro investigation of patulin production under representative conditions in each step of long-term apple storage. Patulin accumulation seemed highly strain dependent and no significant differences between the storage steps were observed. The results also indicated that a high spore inoculum may lead to a strong patulin accumulation even at cold temperatures (1 °C) combined with controlled atmosphere (CA) (3% O2, 1% CO2), suggesting that future control strategies may benefit from considering the duration of storage under CA conditions as well as duration of deck storage.

Download Full-text

Suitability of Cryopreservation for the Long-term Storage of Rare and Endangered Plant Species: a Case History for Cosmos atrosanguineus

Annals of Botany ◽

10.1093/aob/mcg009 ◽

2002 ◽

Vol 91 (1) ◽

pp. 65-74 ◽

Cited By ~ 37

Author(s):

T. WILKINSON

Keyword(s):

Plant Species ◽

Case History ◽

Endangered Plant ◽

Endangered Plant Species ◽

Long Term Storage ◽

Rare And Endangered ◽

Term Storage

Download Full-text

Capture and Purification of Plant Genomic DNA on a Readily-available Cellulose Matrix

10.1101/163212 ◽

2017 ◽

Author(s):

Megan M. Thompson ◽

Estelle M. Hrabak

Keyword(s):

Arabidopsis Thaliana ◽

Nucleic Acids ◽

Plant Species ◽

Efficient Method ◽

Genomic Dna ◽

Low Cost ◽

Long Term Storage ◽

Cellulose Matrix ◽

Term Storage

AbstractWhatman FTA ®Cards are a fast and efficient method for capturing and storing nucleic acids but are cost-prohibitive for some researchers. We developed a method that substitutes readily-available cellulose-based paper and homemade washing buffer for commercial FTA ®Cards and FTA ®Purification Reagent. This method is suitable for long-term storage of DNA from many plant species, including Arabidopsis thaliana, prior to purification and PCR.Method SummaryHere we report a low-cost method for long-term storage of plant genomic DNA on a readily available cellulose matrix.

Download Full-text

Effect of Long-Term Storage on Mycobiota of Barley Grain and Malt

Plants ◽

10.3390/plants10081655 ◽

2021 ◽

Vol 10 (8) ◽

pp. 1655

Author(s):

Soňa Felšöciová ◽

Przemysław Łukasz Kowalczewski ◽

Tomáš Krajčovič ◽

Štefan Dráb ◽

Miroslava Kačániová

Keyword(s):

Barley Grain ◽

In Vitro Assays ◽

Production Cycle ◽

Microscopic Fungi ◽

Microbiome Composition ◽

Long Term Storage ◽

Fungal Microbiome ◽

Term Storage

Contamination of malting barley grain and malt with micromycetes sampled at various periods post-harvest (3rd, 6th, and 9th month of storage) and types of storage (storage silo and floor warehouse) was investigated. Each of these barley grain samples was malted. This article reports on the changes in the fungal microbiome composition and their overall count in barley grain and malt. From the surface-disinfected barley grain samples collected immediately after harvest, there were eight genera isolated, with a predominance of Alternaria. A small increase of isolated microfungi was detected in barley stored in silo for 3 and 6 months (from 142 isolates to 149) and decreased below the number of isolates in barley before storage (133 isolates). Fungal count during storage gradually decreased up to 9 month in barley stored in floor warehouse (from 142 isolates to 84). The initial total count of microscopic fungi in malt before storage was the highest (112 isolates) with 7 genera detected, compared to malts prepared from barley stored for longer time (54 isolates, 7 genera, 9th month of storage). Alternaria was the most abundant and frequent genus. Quantitative representation of the filamentous microscopic fungi was lower compared to yeasts especially in barley and malt prepared from barley stored at third month of storage in both type of storage. Yeasts were identified from all grain samples and malt samples with mass spectrometry. Most attention was given to the widely distributed fungus Penicillium, 79% of strains produced at least one mycotoxin detected under in vitro assays using the TLC method (97% of them produced griseofulvin, 94% CPA, 79% patulin, 14% roquefortin C, and penitrem A was produced by two screening strains under laboratory conditions). It is therefore important to monitor the microflora throughout the production cycle of “barley to beer”.

Download Full-text

Plantlets from encapsulated shoot buds of Catalpa ovata G. Don

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2002.021 ◽

2014 ◽

Vol 71 (3) ◽

pp. 181-186

Author(s):

Halina Wysokińska ◽

Katarzyna Lisowska ◽

Katarzyna Floryanowicz-Czekalska

Keyword(s):

Woody Plant ◽

Storage Period ◽

Shoot Cultures ◽

Shoot Buds ◽

Catalpa Ovata ◽

Alginate Capsules ◽

Woody Plant Medium ◽

The Media ◽

In Vitro Shoot Cultures

Shoot buds isolated from in vitro shoot cultures of Catalpa ovata G. Don were encapsulated using 3% sodium alginate with sucrose (3%) and 50 mM calcium chloride. The morphogenic response of encapsulated buds was affected by such factors, like composition of the media and the presence of growth regulators. The highest frequency of plantlet germination from encapsulated buds (70% within 4 weeks) was obtained on Woody Plant medium (WP) (Lloyd and McCown 1980) containing indole-3-butyric acid (IBA) (1 mg/l). The process was substantially inhibited by cold-storage (4oC) of encapsulated buds. In this case, the frequency response ranged from 3% to 22% dependent on storage period (28 or 42 days) and the presence of the paraffin coat covering the alginate capsules. The plantlets developed from both unstored and stored encapsulated buds of C. ovata were transplanted to soil and grew in pots to phenotypically normal plants.

Download Full-text

Preservation of In Vitro Grown Shoot Tips of Potato (Solanum tuberosum L.) by Different Methods of Cryopreservation

Nepal Journal of Science and Technology ◽

10.3126/njst.v10i0.2804 ◽

1970 ◽

Vol 10 ◽

pp. 15-20

Author(s):

Shambhu P. Dhital ◽

Hira K. Manandhar ◽

Hak T. Lim

Keyword(s):

Sucrose Concentration ◽

Shoot Tips ◽

Efficient Tool ◽

Long Term Storage ◽

In Vitro Plantlets ◽

Tuber Morphology ◽

Vegetatively Propagated Plants ◽

Term Storage

Cryopreservation has been recognized as a practical and efficient tool for long-term storage of vegetatively propagated plants. This study was conducted to investigate the effects of sucrose concentration, hardening temperature and different cryopreservation methods on the survival rate of potato shoot tips after cryopreservation. Excised shoot tips of in vitro plantlets of potato cultivars, Atlantic and Superior were cryopreserved by vitrification, encapsulationvitrification and encapsulation-dehydration. Cryopreservation by vitrification method was used to determine the optimum concentration of sucrose and cold hardening temperature during sub-culturing period to the donor plantlets. Nine-percent sucrose gave 46.7% survival in Atlantic and 40% in Superior. The most optimum hardening temperature for 50% survival in Atlantic and 43.3% in Superior was 10°C. In the case of comparative study of three different cryopreservation methods, the highest survival (52%) as well as regeneration (46%) were observed when the shoot tips were cryopreserved by encapsulation-vitrification method, and the lowest survival (36%) and regeneration (28%) from the vitrification. Plant and tuber morphology of potato regenerated after cryopreservation were similar to those of the non-cryopreserved in vitro plantlets (control). Thus, this study demonstrated that encapsulation-vitrification method was the most effective one among other methods for higher survival as well as regeneration in in vitro shoot tips of potato.Key words: Cryopreservation; Dehydration; Encapsulation; Potato; Regeneration; VitrificationDOI: 10.3126/njst.v10i0.2804Nepal Journal of Science and Technology Volume 10, 2009 December Page: 15-20

Download Full-text

Cryopreservation of cherry rootstock Gisela 5 using vitrification procedure

Horticultural Science ◽

10.17221/234/2013-hortsci ◽

2014 ◽

Vol 41 (No. 2) ◽

pp. 55-63 ◽

Cited By ~ 2

Author(s):

Dj. Ružić ◽

T. Vujović ◽

R. Cerović

Keyword(s):

Sucrose Concentration ◽

Survival Rates ◽

Shoot Tips ◽

Prunus Cerasus ◽

Ms Medium ◽

Long Term Storage ◽

Term Storage ◽

Gisela 5

In vitro-grown shoot tips of Gisela 5 (Prunus cerasus × Prunus canescens) cherry rootstock were tested for regrowth after cryopreservation using vitrification technique. Explants were precultured in the dark at 23°C, in a liquid MS medium with a progressively increasing sucrose concentration (0.3 M for 15 h, then 0.7 M for 5 h), and subsequently loaded in a solution containing 2 M glycerol and 0.4 M sucrose for 20 minutes. Shoot tips were dehydrated at 0°C using either the original PVS2 or modified PVS2 solution (PVS A3 – 22.5% sucrose, 37.5% glycerol, 15% ethylene glycol and 15% DMSO) for 30, 40 and 50 minutes. The survival and regrowth of the cryopreserved shoot tips dehydrated with the original PVS2 solution ranged between 36–54% and 8–17%, respectively. However, the dehydration with the PVS A3 solution resulted in considerably higher survival rates (81–92%), as well as higher regrowth rates (39–56%) after cryopreservation. These results prove the feasibility of the PVS A3-based vitrification technique for a long-term storage of this genotype.  

Download Full-text