Evaluation ofin vitrogenotoxic effects induced byin vitroanther culture conditions in sunflower

Abstract

Sunflower is a globally important oilseed, food, and ornamental crop. This study seeks to investigate the genotoxic effects of tissue culture parameters in sunflower calli tissues belongs to two genotypes obtained via anther culture. Anthers were pretreated with cold for 24 hours at 4°C and heat for 2 days at 35°C in the dark and plated onto media supplemented with different concentrations and combinations of 6-benzylaminopurine, 2,4-dichlorophenoxyacetic acid, α-naphthalene acetic acid and indole-3-acetic acid. Obtaining calli tissues were used to detect the DNA damage levels by Comet assay, evaluating changes on superoxide dismutase and guaiacol peroxidase activities derived from in vitro culture factors. 0.5 mg/L 2,4-dichlorophenoxyacetic acid and 2 mg/L α-naphthalene acetic acid from plant growth regulators showed acute genotoxic effect while 0.5 mg/L indole-3-acetic acid and 0.5 mg/L α-naphthalene acetic acid showed no genotoxic effect. Total protein content analysis of antioxidant enzymes revealed that although superoxide dismutase activity did not increase, Guaiacol peroxidase (GPOX) activity decreased in comparison to control. The obtained results have indicated that in vitro culture factors apparently lead to genotoxicity and oxidative stress.

Origanum vulgare L. Cuttings Rhizogenesis in Microclonal Reproduction in Vitro

“…Загалом для оптимізації етапу укорінення рослин-регенерантів поєднання використання різних видів регуляторів росту, варіювання їх концентрацій широко вивчається в культурі in vitro різноманітних видів рослин, у т.ч. і лікарських [20], застосовується як для оптимізації біотехнологій мікроклонального розмноження цінних видів рослин, так і з використанням калюсних культур, культури пиляків тощо для біотехнологічного забезпечення селекційного процесу [21,22]. Утім дослідники наголошують на диференційованій генотиповій реакції за рахунок внутрішньовидової спадкової мінливості на варіювання як фітогормонального складу живильних середовищ, так і взагалі умов культивування [23,24].…”

Determination of genotoxic damages of picloram and dicamba with comet assay in Allium cepa rooted in tissue culture and distilled water

“…It is currently used to directly determine DNA damages and repair in both animal and plant materials [17][18][19][20][21][22]. There are several model species like Arabidopsis thaliana, Vicia faba, Nicotiana tabacum or Allium cepa to perform Comet test [20,23]. Allium cepa is the most widely preferred plant species and has been used in genotoxicity studies related to many chemical agents [19,21,24,25].…”

Understanding In Vitro Tissue Culture-Induced Variation Phenomenon in Microspore System

“…A global DNA hypomethylation during the change of the gametophytic to sporophytic fate and first embryogenic divisions were observed in barley [ 100 ]. Among inductive stressful conditions, darkness is one of the most exploited in cereals [ 101 ]. When mannitol is applied, triggering microspores to dividing [ 73 ], the culture is subjected to osmotic, and carbon starvation discussed earlier [ 102 ].…”

“…The tissue culture medium itself may be treated as a stressful factor. The regeneration medium contains many ingredients such as 6-benzylaminopurine, 2,4-dichlorophenoxyacetic acid, alpha-naphthalene acetic acid, and indole-3-acetic acid not necessary are neutral to the cells may induce TCIV [ 101 , 106 , 107 ]. As such ingredients may be toxic, they may lead to genomic DNA degradation as indicated by comet analysis [ 101 ].…”

“…The regeneration medium contains many ingredients such as 6-benzylaminopurine, 2,4-dichlorophenoxyacetic acid, alpha-naphthalene acetic acid, and indole-3-acetic acid not necessary are neutral to the cells may induce TCIV [ 101 , 106 , 107 ]. As such ingredients may be toxic, they may lead to genomic DNA degradation as indicated by comet analysis [ 101 ]. It was also demonstrated that copper and silver ions containing media might also affect DNA methylation patterns [ 12 ], resulting in DNA sequence changes [ 62 ].…”