Comet and cytogenetic tests as tools for evaluating genomic instability in seeds of Oryza sativa L. and Phaseolus vulgaris L. from gene banks

Dantas, Alisson F.; Lopes, Renato Matos; Fascineli, Maria Luiza; José, Solange Carvalho Barrios Roveri; Pádua, Juliano Gomes; Gimenes, Marcos Aparecido; Grisólia, César Koppe

doi:10.1590/1678-4685-gmb-2017-0020

Cited by 8 publications

(4 citation statements)

References 34 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Other studies used these buffers supplemented with Na 2 EDTA and with or without DMSO [ 24 , 28 , 51 ]. There are also variations in the composition of the lysis solution used to disrupt the nuclear membrane and protein components [ 23 , 27 , 30 , 43 , 44 ]. Several reports concluded, however, that the lysis step is redundant [ 20 , 21 , 52 ].…”

Section: Discussionmentioning

confidence: 99%

“…An increasing number of studies, however, have revealed that DNA damage and repair capacity are also major factors that determine germplasm viability [ 40 ]. Indeed, the loss of genomic integrity is one of the first stages of germplasm deterioration [ 30 ]. Understanding the mechanism of DNA damage and repair, and the development of a method to assess and monitor genome integrity, will improve the application and reliability of ex-situ preservation of plant genetic resources in gene banks.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

Oxidative damage and DNA repair in desiccated recalcitrant embryonic axes of Acer pseudoplatanus L.

et al. 2022

View full text Add to dashboard Cite

Background Most plants encounter water stress at one or more different stages of their life cycle. The maintenance of genetic stability is the integral component of desiccation tolerance that defines the storage ability and long-term survival of seeds. Embryonic axes of desiccation-sensitive recalcitrant seeds of Acer pseudoplatnus L. were used to investigate the genotoxic effect of desiccation. Alkaline single-cell gel electrophoresis (comet assay) methodology was optimized and used to provide unique insights into the onset and repair of DNA strand breaks and 8-oxo-7,8-dihydroguanine (8-oxoG) formation during progressive steps of desiccation and rehydration. Results The loss of DNA integrity and impairment of damage repair were significant predictors of the viability of embryonic axes. In contrast to the comet assay, automated electrophoresis failed to detect changes in DNA integrity resulting from desiccation. Notably, no significant correlation was observed between hydroxyl radical (٠OH) production and 8-oxoG formation, although the former is regarded to play a major role in guanine oxidation. Conclusions The high-throughput comet assay represents a sensitive tool for monitoring discrete changes in DNA integrity and assessing the viability status in plant germplasm processed for long-term storage.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Oxidative damage and DNA repair in desiccated recalcitrant embryonic axes of Acer pseudoplatanus L.

et al. 2022

View full text Add to dashboard Cite

show abstract

“…However, under the influence of environmental and anthropogenic factors, the areas of their growth are decreasing (Schlaepfer et al, 2018;Swift et al, 2019;González et al, 2020;Mayerhofer et al, 2021). In modern genebanks, to preserve plant genetic resources, along with traditional methods, biotechnological ones are successfully used, which include preserving plant germplasm in culture in vitro and cryopreservation at a temperature of -196°C (Dantas et al, 2018;Popova, 2018;Sofi et al, 2020;Özbek and Zencirci, 2021). Cryopreservation of germplasm of plant material in liquid nitrogen guarantees long-term preservation of the material and at the same time reduces the availability of the occurrence of genetic changes possible in in vitro culture at positive temperatures (Jenderek and Reed, 2017;Kalaiselvi et al, 2017;Kaya et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Micropropagation of berry crops for creation of germplasm cryobanks

et al. 2024

View full text Add to dashboard Cite

One of the main stages of cryopreservation of meristematic tissues in vegetative plants is a clonal micropropagation, which includes isolating the explants of the raw material in vitro and optimizing the culture medium for micropropagation. As the result of our studies, the optimal periods for in vitro micropropagation are: first - isolation of explants from initiated shoots of dormant buds (blackcurrants and raspberries) in January-March; the second - from actively growing shoots (blackcurrants and raspberries) in May-June, from the formed mustache (strawberry) in July-August. The optimal drugs for sterilization of raspberry explants are: a) 0.1% HgCl2 (6 min), then 3% H2O2 (15 min); b) chlorine-containing bleach «Domestos» in the dilution of H2O 1:9 (10 min). For blackcurrant: a) 0.1% HgCl2 (5 min) in combination with 0.1% fungicide “Topaz” (30 min); b) 0.1% HgCl2 (5 min) in combination with the treatment with KMnO4 (30 min); c) “Domestos” in the dilution of H2O 1:5 (20 min). For strawberry: a) 0.1% HgCl2 (6 min) followed by treatment with 3% H2O2 10 (min); b) 1% deochlor (7 min), 3% H2O2 (10 min); c) “Domestos” in the dilution of H2O 1:5 (8 min) with subsequent treatment 0,1% HgCl2 -7 min, then 0,20 mg/l КМnO4 - 30 min. Optimal compositions of culture media for micropropagation of blackcurrant - Murashige and Skoog (MS) medium with 0.5 mg L-1 BAP, 0.5 mg L-1 GA3, 0.1 mg L-1 IBA and 20 g L-1 glucose. For raspberry -MS medium with 0.5 mg L-1 BAP, 0.1 mg L-1 IBA, 10 mg L-1 iron chelate and 30 g L-1 sucrose. For strawberry - MS medium with 0.3 mg L-1 BAP, 0.01 mg L-1 IBA, 0.2 mg L-1 GA3, 10 mg L-1 iron chelate and 30 g L-1 sucrose. Based on these studies, the cryobank was created, which include the germplasm of in vitro meristematic tissues in 66 cultivars, hybrids and wild-growing forms of blackcurrant, raspberry and strawberry. Therefore, the aim of the research was to obtain aseptic plants, clonal micropropagation and the creation of a cryogenic collection of germplasm based on the developed technology.

show abstract

“…The same approach was also used by de Sousa Araujo et al ( 2019 ) to disclose the genotoxic impact of prolonged kinetin-based seed priming in M. truncatula . Genomic instability was evaluated in seeds of rice and beans stored in gene banks using comet assay to investigate the molecular bases of seed deterioration (Dantas et al 2018 ). Comet assay has been tested on embryos or embryo axes isolated from eggplant (Kiran et al 2020 ), M. truncatula (Pagano et al 2022a ), and Acer pseudoplatanus L. (Plitta-Michalak et al 2022 ) seeds.…”

Section: Introductionmentioning

confidence: 99%

Molecular dynamics of seed priming at the crossroads between basic and applied research

Pagano¹,

Macovei²,

Balestrazzi³

2023

Plant Cell Rep

View full text Add to dashboard Cite

Key message The potential of seed priming is still not fully exploited. Our limited knowledge of the molecular dynamics of seed pre-germinative metabolism is the main hindrance to more effective new-generation techniques. Abstract Climate change and other recent global crises are disrupting food security. To cope with the current demand for increased food, feed, and biofuel production, while preserving sustainability, continuous technological innovation should be provided to the agri-food sector. Seed priming, a pre-sowing technique used to increase seed vigor, has become a valuable tool due to its potential to enhance germination and stress resilience under changing environments. Successful priming protocols result from the ability to properly act on the seed pre-germinative metabolism and stimulate events that are crucial for seed quality. However, the technique still requires constant optimization, and researchers are committed to addressing some key open questions to overcome such drawbacks. In this review, an update of the current scientific and technical knowledge related to seed priming is provided. The rehydration–dehydration cycle associated with priming treatments can be described in terms of metabolic pathways that are triggered, modulated, or turned off, depending on the seed physiological stage. Understanding the ways seed priming affects, either positively or negatively, such metabolic pathways and impacts gene expression and protein/metabolite accumulation/depletion represents an essential step toward the identification of novel seed quality hallmarks. The need to expand the basic knowledge on the molecular mechanisms ruling the seed response to priming is underlined along with the strong potential of applied research on primed seeds as a source of seed quality hallmarks. This route will hasten the implementation of seed priming techniques needed to support sustainable agriculture systems.

show abstract

Comet and cytogenetic tests as tools for evaluating genomic instability in seeds of Oryza sativa L. and Phaseolus vulgaris L. from gene banks

Cited by 8 publications

References 34 publications

Oxidative damage and DNA repair in desiccated recalcitrant embryonic axes of Acer pseudoplatanus L.

Oxidative damage and DNA repair in desiccated recalcitrant embryonic axes of Acer pseudoplatanus L.

Micropropagation of berry crops for creation of germplasm cryobanks

Molecular dynamics of seed priming at the crossroads between basic and applied research

Contact Info

Product

Resources

About