scholarly journals Triticale Green Plant Regeneration Is Due to DNA Methylation and Sequence Changes Affecting Distinct Sequence Contexts in the Presence of Copper Ions in Induction Medium

Cells ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 84
Author(s):  
Renata Orłowska ◽  
Katarzyna Anna Pachota ◽  
Piotr Androsiuk ◽  
Piotr Tomasz Bednarek
Keyword(s):  
Plant Regeneration ◽  
Anther Culture ◽  
Sequence Variation ◽  
De Novo ◽  
Copper Ions ◽  
Green Plant ◽  
Dna Demethylation ◽  
Induction Medium ◽  
Regeneration Efficiency ◽  
De Novo Methylation

Metal ions in the induction medium are essential ingredients allowing green plant regeneration. For instance, Cu(II) and Ag(I) ions may affect the mitochondrial electron transport chain, influencing the Yang cycle and synthesis of S-adenosyl-L-methionine, the prominent donor of the methylation group for all cellular compounds, including cytosines. If the ion concentrations are not balanced, they can interfere with the proper flow of electrons in the respiratory chain and ATP production. Under oxidative stress, methylated cytosines might be subjected to mutations impacting green plant regeneration efficiency. Varying Cu(II) and Ag(I) concentrations in the induction medium and time of anther culture, nine trials of anther culture-derived regenerants of triticale were derived. The methylation-sensitive AFLP approach quantitative characteristics of tissue culture-induced variation, including sequence variation, DNA demethylation, and DNA de novo methylation for all symmetric-CG, CHG, and asymmetric-CHH sequence contexts, were evaluated for all trials. In addition, the implementation of mediation analysis allowed evaluating relationships between factors influencing green plant regeneration efficiency. It was demonstrated that Cu(II) ions mediated relationships between: (1) de novo methylation in the CHH context and sequence variation in the CHH, (2) sequence variation in CHH and green plant regeneration efficiency, (3) de novo methylation in CHH sequences and green plant regeneration, (4) between sequence variation in the CHG context, and green plant regeneration efficiency. Cu(II) ions were not a mediator between de novo methylation in the CG context and green plant regeneration. The latter relationship was mediated by sequence variation in the CG context. On the other hand, we failed to identify any mediating action of Ag(I) ions or the moderating role of time. Furthermore, demethylation in any sequence context seems not to participate in any relationships leading to green plant regeneration, sequence variation, and the involvement of Cu(II) or Ag(I) as mediators.

scholarly journals Time of In Vitro Anther Culture May Moderate Action of Copper and Silver Ions that Affect the Relationship between DNA Methylation Change and the Yield of Barley Green Regenerants

Plants ◽  
2020 ◽  
Vol 9 (9) ◽  
pp. 1064
Author(s):  
Piotr T. Bednarek ◽  
Renata Orłowska
Keyword(s):  
Anther Culture ◽  
De Novo ◽  
Silver Ions ◽  
Dna Demethylation ◽  
Induction Medium ◽  
Green Plants ◽  
Ion Concentrations ◽  
Regenerated Plants ◽  
Short Time

Plant anther culture allows for the regeneration of uniform and homozygous double haploids. However, off-type regenerants may appear as a result of so-called tissue culture-induced variation (TCIV). In addition, the presence of Cu2+ and Ag+ ions in the culture medium might influence the number of green plants. The regenerants were obtained via anther cultures of barley under varying Cu2+ and Ag+ ion concentrations in the induction medium during distinct time conditions. DArTseqMet markers were evaluated based on regenerants and donor plants and delivering data on DNA demethylation (DM) and de novo methylation (DNM) and changes in methylation (Delta). The number of green regenerated plants per 100 anthers (GPs) was evaluated. The Cu2+ and Ag+ ion concentrations moderated relationships between Delta and the number of green plants conditional on time of tissue cultures. Depending on the ions, moderated moderation is valid within the different time of anther culture. When the highest concentration of copper is analyzed, plant regeneration is possible under short ‘Time’ (21 days) of anther culture wherein Delta is negative or under elongated Time when Delta is positive. Under 21 days of culture, the highest concentration of silver ions and when Delta is negative, some regenerants could be evaluated. However, under high Ag+ concentration when Time of culture is long and Delta positive, the highest number of green plants could be obtained.

scholarly journals Structural Equation Modeling (SEM) Analysis of Sequence Variation and Green Plant Regeneration via Anther Culture in Barley

Cells ◽  
2021 ◽  
Vol 10 (10) ◽  
pp. 2774
Author(s):  
Piotr Tomasz Bednarek ◽  
Renata Orłowska ◽  
Dariusz Rafał Mańkowski ◽  
Sylwia Oleszczuk ◽  
Jacek Zebrowski
Keyword(s):  
Dna Methylation ◽  
Structural Equation Modeling ◽  
Plant Regeneration ◽  
Anther Culture ◽  
Structural Equation ◽  
Sequence Variation ◽  
Green Plant ◽  
Complex Model ◽  
Equation Modeling ◽  
Green Plants

The process of anther culture involves numerous abiotic stresses required for cellular reprogramming, microspore developmental switch, and plant regeneration. These stresses affect DNA methylation patterns, sequence variation, and the number of green plants regenerated. Recently, in barley (Hordeum vulgare L.), mediation analysis linked DNA methylation changes, copper (Cu2+) and silver (Ag+) ion concentrations, sequence variation, β-glucans, green plants, and duration of anther culture (Time). Although several models were used to explain particular aspects of the relationships between these factors, a generalized complex model employing all these types of data was not established. In this study, we combined the previously described partial models into a single complex model using the structural equation modeling approach. Based on the evaluated model, we demonstrated that stress conditions (such as starvation and darkness) influence β-glucans employed by cells for glycolysis and the tricarboxylic acid cycle. Additionally, Cu2+ and Ag+ ions affect DNA methylation and induce sequence variation. Moreover, these ions link DNA methylation with green plants. The structural equation model also showed the role of time in relationships between parameters included in the model and influencing plant regeneration via anther culture. Utilization of structural equation modeling may have both scientific and practical implications, as it demonstrates links between biological phenomena (e.g., culture-induced variation, green plant regeneration and biochemical pathways), and provides opportunities for regulating these phenomena for particular biotechnological purposes.

scholarly journals Impact of Ionic Liquids on Induction of Wheat Microspore Embryogenesis and Plant Regeneration

Agronomy ◽  
2020 ◽  
Vol 10 (6) ◽  
pp. 839
Author(s):  
Dorota Weigt ◽  
Idzi Siatkowski ◽  
Magdalena Magaj ◽  
Agnieszka Tomkowiak ◽  
Jerzy Nawracała
Keyword(s):  
Ionic Liquids ◽  
Plant Regeneration ◽  
Positive Impact ◽  
Microspore Embryogenesis ◽  
Green Plant ◽  
Induction Medium ◽  
Dichlorophenoxyacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
The Impact

Ionic liquids are novel compounds with unique chemical and physical properties. They can be received based on synthetic auxins like 2,4-dichlorophenoxyacetic acid or dicamba, which are commonly used hormones in microspore embryogenesis. Nevertheless, ionic liquids have not been adapted in plant in vitro culture thus far. Therefore, we studied the impact of ionic liquids on the ability to undergo microspore embryogenesis in anther cultures of wheat. Two embryogenic and two recalcitrant genotypes were used for this study. Ten combinations of ionic liquids and 2,4-dichlorophenoxyacetic acid were added to the induction medium. In most cases, they stimulated induction of microspore embryogenesis and green plant regeneration more than a control medium supplemented with only 2,4-dichlorophenoxyacetic acid. Two treatments were the most favorable, resulting in over two times greater efficiency of microspore embryogenesis induction in comparison to the control. The effect of breaking down the genotype recalcitrance (manifested by green plant formation) was observed under the influence of 5 ionic liquids treatments. Summing up, ionic liquids had a positive impact on microspore embryogenesis induction and green plant regeneration, increasing the efficiency of these phenomena in both embryogenic and recalcitrant genotypes. Herbicidal ionic liquids can be successfully used in in vitro cultures.

scholarly journals ROLE OF POLYAMINES IN INHIBITION OF ETHYLENE BIOSYNTHESIS AND THEIR EFFECTS ON RICE ANTHER CULTURE DEVELOPMENT

2016 ◽  
Vol 9 (2) ◽  
pp. 60 ◽  
Author(s):  
Iswari S. Dewi ◽  
Bambang S. Purwoko
Keyword(s):  
Plant Regeneration ◽  
Anther Culture ◽  
Culture Media ◽  
Callus Formation ◽  
Indica Rice ◽  
Ethylene Biosynthesis ◽  
Green Plant ◽  
Culture Development ◽  
Acc Content

The polyamines such as putrescine, spermidine, and spermine were reported to increase green plant regeneration in rice anther culture. Low response of anther culture of rice sub-species indica may be improved with the addition of putrescine in the culture media. Four experiments were conducted to study the role of polyamines in inhibition of ethylene biosynthesis and their effects on rice anther culture development. Anthers of two subspecies of rice, indica (IR64, Krowal, Jatiluhur) and japonica (Taipei 309) were cultured onto media supplemented with putrescine (N6P) and without putrescine (N6). Young<br />panicles containing the anthers at mid-to-late nucleate microspores were cold pretreated at 5 + 2°C and incubated in the dark for 8 days before the anthers were cultured. Results<br />showed that medium without putrescine produced an earlier senescence of indica rice anther than that of japonica. The addition of 10-3 M putrescine into the culture media inhibited ethylene biosynthesis as anther senescence delayed, increased the three polyamines contents, and decreased the ACC content as well as ACC oxydase activity in anther-derived calli. In the anther and anther-derived calli of subspecies indica, the total<br />polyamines content was lower (10.14 nM g-1 anther and 8.48 nM g-1 calli) than that of subspecies japonica (12.61 nM g-1 anther and 10.16 nM g-1 calli), whereas the ethylene production was higher (32.31 nM g-1 anther and 2.48 nM g-1 calli) than the japonica (31.68 nM g-1 anther and 1.76 nM g-1 calli). This study suggests that application of 10-3 M putrescine in anther culture of rice subspecies indica improves androgenesis by inhibiting<br />early senescence of cultured anthers and enhancing embryo or callus formation from microspores.

scholarly journals Efficiency of anther culture technique in the production of wheat double haploids

2008 ◽  
pp. 35-40 ◽  
Author(s):  
Ankica Kondic-Spika ◽  
Borislav Kobiljski ◽  
Nikola Hristov
Keyword(s):  
Triticum Aestivum ◽  
Plant Regeneration ◽  
Anther Culture ◽  
Triticum Aestivum L ◽  
Green Plant ◽  
Culture Technique ◽  
Average Yield ◽  
Green Plants ◽  
Double Haploids

The objective of the study was to investigate efficiency of anther culture in the production of spontaneous double haploids from randomly selected heterozygous genotypes of wheat (Triticum aestivum L.). Anthers of 20 F1 wheat combinations were grown in vitro on a modified Potato-2 medium. All of the examined genotypes have shown the ability to produce pollen calluses as well as to regenerate green plants. On average for the whole experiment material, 47.2 calluses were produced per 100 cultured anthers. The green plant regeneration ranged from 0.8 to 13.4 green plants per spike, with an overall mean of 5.8. From the total of 582 regenerated green plants, 47.9% (279) were spontaneous double haploids. The final average yield from the study was 2.8 double haploids per spike.

Altered chromatin condensation of heat-stressed spermatozoa perturbs the dynamics of DNA methylation reprogramming in the paternal genome after in vitro fertilisation in cattle

2014 ◽  
Vol 26 (8) ◽  
pp. 1107 ◽  
Author(s):  
Mohammad Bozlur Rahman ◽  
Md. Mostofa Kamal ◽  
Tom Rijsselaere ◽  
Leen Vandaele ◽  
Mohammed Shamsuddin ◽  
...  
Keyword(s):  
Dna Methylation ◽  
De Novo ◽  
Chromatin Condensation ◽  
Dna Demethylation ◽  
In Vitro Fertilisation ◽  
Paternal Genome ◽  
Time Points ◽  
Fertilisation Rate ◽  
De Novo Methylation

Shortly after penetration of the oocyte, sperm DNA is actively demethylated, which is required for totipotent zygotic development. Aberrant DNA methylation is thought to be associated with altered chromatin condensation of spermatozoa. The objectives of this study were to investigate the dynamics of DNA methylation reprogramming in the paternal pronucleus and subsequent fertilisation potential of heat-stressed bull spermatozoa having altered chromatin condensation. Hence, bovine zygotes (n = 1239) were collected at three different time points (12, 18 and 24 h post insemination, hpi), and stained with an antibody against 5-methylcytosine. Fluorescence intensities of paternal and maternal pronuclei were measured by ImageJ. DNA methylation patterns in paternal pronuclei derived from heat-stressed spermatozoa did not differ between time points (P > 0.05), whereas control zygotes clearly showed demethylation and de novo methylation at 18 and 24 hpi, respectively. Moreover, heat-stressed spermatozoa showed a highly reduced (P < 0.01) fertilisation rate compared with non-heat-stressed or normal control spermatozoa (53.7% vs 70.2% or 81.5%, respectively). Our data show that the normal pattern of active DNA demethylation followed by de novo methylation in the paternal pronucleus is perturbed when oocytes are fertilised with heat-stressed spermatozoa, which may be responsible for decreased fertilisation potential.

Sign in / Sign up

Export Citation Format

Share Document