scholarly journals Anther Culture Effectiveness in Producing Doubled Haploids of Cereals/ Putekðòu Kultûras Efektivitâte Graudaugu Dubultoto Haploîdu Izveidoðanâ

Author(s):  
Dace Grauda ◽  
Andra Miķelsone ◽  
Nadežda Ļisina ◽  
Kârlis Žagata ◽  
Reinis Ornicāns ◽  
...  

Abstract Our goal was to improve the method of obtaining double haploids (DH) by anther culture from Latvian breeding material for several agriculturally important cereals in Latvia: barley, spring and winter wheat. Hybrids from Latvian breeders were initial material. It was found that copper (2.5 mg/l) added to barley anther pre-treatment media and to both barley (C3) and wheat (liquid AMC) induction media, resulted in a higher percentage of green plants-regenerants. In general, the winter wheat hybrids exhibited lower embryogenesis than spring wheat hybrids. Obtaining a large number of DH lines from barley and wheat hybrids with unknown androgenesis response can be organized in two stages: first - selection from breeding initial material hybrids responsive in anther culture, which enables production of green plants-regenerants, and second - production of DH lines in a large scale from selected hybrids.

2020 ◽  
Vol 56 (No. 4) ◽  
pp. 150-158
Author(s):  
Osama Zuhair Kanbar ◽  
Csaba Lantos ◽  
Paul Karumba Chege ◽  
Erzsébet Kiss ◽  
János Pauk

We investigated the anther culture (AC) efficiency of thirteen F<sub>4</sub> combinations of winter wheat (Triticum aestivum L.). The genotype dependency was assessed during the induction of the androgenic entities, i.e. embryo-like structures (ELS), regenerated-, green-, albino-, and transplanted plantlets. The number of green plantlets per 100 anthers (GP/100A) varied from 0.36 to 24.74 GP/100A with a mean of 8.31 GP/100A. Albino plantlets (AP) occurred in each combination, ranging from 0.20 to 22.80 AP/100A with an average value of 5.59 AP/100A. Between 25–87.76 doubled haploid (DH) plants per 100 acclimatised plantlets (DH/100ADP), depending on the combination, with a mean of 59.74% were recovered. We have found the highest DH production in the combinations Béres/Midas, Kalász/Tacitus, Béres/Pamier, and Premio/5009. This improves remarkably the choice of basic genetic material in subsequent crossing programmes. These observations emphasise the usability and efficiency of in vitro AC in producing a large number of DH lines for breeding and the applied researches of winter wheat. Although albinism was found in each combination, it was mitigated by the in vitro AC application.


2018 ◽  
Vol 22 ◽  
pp. 252-256 ◽  
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
M. S. Boyko

Aim. Testing the haploproduction ability of 47 varieties and 143 F1 hybrids of winter bread wheat. Methods. In vitro culture of isolated anthers of wheat. The percentage of callus and regeneration of green plants for each genotype calculated as a percentage of the planted anthers. Results. The differences in the frequency of сallus induction and the ability to regenerate plants in the process of androgenesiss in vitro of winter soft wheat were detected. The range of variation haploproduction activiti was broad. The sign of "the formation of сallus" for varieties was in limited from 0 to 21.2 % and for hybrids – from 0 to 38.8 %; on the sign of "regeneration of green plants" for varieties – from 0 to 9.4 %, and for hybrids from 0 to 9.1 %. The positive effect of 1BL/1RS translocation on signs of haploproduction in anther culture Triticum aestivum L. Conclusions. Genotype-specific of microspores morphogenetic reactions of soft winter wheat in the process of androgenesis in vitro were revealed. It has been found that F1 hybrids have a higher potential production potential in anther culture than varieties. During the period from 2011 to 2017, the 580 lines of winter wheat were transferred to the breeding departments. Keywords: soft winter wheat, anther culture in vitro, callus, regeneration.


Author(s):  
Mridhu Sharma ◽  
A. K. Singh ◽  
R. K. Salgotra ◽  
Manmohan Sharma ◽  
Bupesh Ku. Sharma ◽  
...  

With different culture conditions and concentrations of growth regulators and supplements, Anther culture technique can be easily employed for the production of haploids under in vitro conditions. Aims: The present study was undertaken with the objective to optimize the development of doubled haploids using anthers for in vitro induction of callus on N6 medium. Place and Duration of Study: The samples (BC2F1 seeds) were raised previously in Skuast-J. From total degree program of 3 years, this work related to tissue culture technique was done in one year from January 2018 to January 2019. Methodology: The effect of levels of 2, 4-dichlorophenoxy acetic acid (2, 4-D) i.e. 0 to 3 mg/L in basal N6 media was observed on callus induction frequency (CIF). The effect of duration of cold pre-treatment was observed on callus induction frequency at 2.5 mg/L of 2, 4-D by giving the cold pre-treatment at 4ºC from 8 to 12 days. Also the effect of different amino acids was checked on callus induction frequency. Results: Highest callus induction frequency of 9.39 per cent was observed in N6 medium fortified with 2.5 mg/L 2, 4-D and lowest callus induction frequency of 2.52 per cent at the concentration of 1.0 mg/L. The cold pre-treatment for 10 days gave highest callus induction frequency of 1.44 per cent and lowest callus induction frequency of 0.44 per cent was obtained at cold pre treatment for 8 days. The highest callus induction frequency of 12.55 per cent was observed in case of media supplemented with 25 mg/L tryptophan and 40 mg/L cysteine and lowest callus induction frequency of 7.18 per cent was observed when media was supplemented with 560 mg/L proline. Conclusion: The cold pre-treatment of 10 days at 4ºC on media supplemented with 2.5 mg/L of 2, 4-D and combination of 25 mg/L tryptophan and 40 mg/L cysteine proves to provide best androgenesis conditions for anthers from BC2F1 population.


2019 ◽  
Vol 2 (2) ◽  
pp. 24-32
Author(s):  
M. E. Mikhailov

The doubled haploid (DH) lines, obtained by doubling the haploid genome, are now widely used in breeding many crops, since they allow to transfer gene variants to the homozygous state in a short time. However, the advantages of doubled haploids are not fully utilized in maize breeding. The present work is devoted to the evaluation of the backcrossing method efficiency and to further development of the original schemes of creating highly productive homozygous maize lines on the basis of DH lines originating from an interline F1 hybrid. Rf7 and Ku123 maize lines were used as the initial material. The breeding cycle consisted of producing haploid plants in the selected genotype (matroclinic haploidy using an inducer), subsequent chromosome doubling (colchicine‑induced or spontaneous), followed by multiplication of the doubled haploids for obtaining a new set of DH lines. In the first cycle, the DH lines were obtained from the F1 hybrid (Rf7 × Ku123), while in the subsequent cycles they were obtained from the genotypes obtained by crossing a DH line selected from the previous cycle with F1, P1 or P2. Three cycles of selection for productivity were performed, and in 2017 the DH lines obtained in all cycles were simultaneously tested in the field. The breeding progress was estimated by the increase in the first ear productivity compared to the best parent Rf7 (103.9 g per plant in 2017). The first selection cycle resulted in 43 DH lines obtained on the basis of the F1 hybrid. Productivity of the best line rk‑5 amounted to 112.5 g per plant. Three lines (rk‑6, rk‑5 and rk‑22) selected for the next cycle were further crossed with F1 or with the parental line Rf7. The second selection cycle yielded three series containing 41, 49 and 16 lines, while productivity of the best genotypes was 121.2, 117.0 и 107.1 g per plant, respectively. The third cycle included populations of 24 and 8 lines obtained through backcrosses with Rf7 and Ku123 lines, respectively. The best genotypes in these series had productivity of 135.6 and 97.7 g per plant. As a result of selection, the obtained rk‑433 line had a productivity 30.5% higher than that of the best parent Rf7. The progress averaged 10.2% per cycle. In maize breeding using doubled haploids it is promising to use backcrosses of the selected DH lines with the initial material or with F1. Thanks to such an approach, a noticeable progress can be reached with a small number of cycles including from 20 to 50 DH lines.


Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
M. S. Boiko ◽  
H. O. Dobrova ◽  
S. V. Ahafonova

Aim. To estimate the level of in vitro haploproduction androgenic ability of different generations populations (F1, F2) soft winter wheat hybrids of the same combination to maximize the use of biotechnology cereals. Methods. Obtaining of bread wheat double haploid lines by anther culture in vitro. The statistical methods. Results. The influence different generation of wheat hybrids on the processes of induction and regeneration in anther culture of winter bread wheat were studied. The 21 green plants-regenerants were received. Conclusions. The using of anthers of the second generation simple hybrids as donor material for the effective technology creation of doubled haploid of bread winter wheat was shown.Keywords: wheat, anther culture in vitro, callus, regeneration, generation simple hybrids.


2016 ◽  
Vol 58 (1) ◽  
pp. 95-102 ◽  
Author(s):  
Theano Lazaridou ◽  
Chryssanthi Pankou ◽  
Ioannis Xynias ◽  
Demetrios Roupakias

AbstractThe present study was conducted to determine the effect of the D genome on embryoid induction and green plant regeneration in wheat anther culture and how it is influenced by low temperature and mannitol treatment. For this reason, the anther culture response of two Canadian bread wheat cultivars and their extracted tetraploids (AABB) was studied. As controls two cultivars well responding to anther-culture (i.e. cvs. Kavkaz/Cgn and Acheron) and a no-responding cultivar (cv. Vergina) were used. Approximately 3000 anthers of these cultivars were cultured and three pre-treatments were applied: cold pre-treatment for 7 and 18 days at 4°C, and 0.3M mannitol for seven days at 4°C. W14 and 190-2 were used as induction and regeneration media, respectively, and the basic MS medium as the rooting medium. No green plants were produced from the tetraploids, which supports the view that the D-genome chromosomes are necessary for androgenic response in wheat. Furthermore, the Canadian cultivars performed better after 18-day pre-treatment at 4°C. The extracted tetraploids produced fewer embryoids and performed better after seven days of cold pre-treatment. The controls well responding to anther culture performed better than the Canadian cultivars, although their best response was recorded after seven-day cold pre-treatment. Cultivar Vergina produced no green plants. The presence of mannitol influenced negatively both embryoid and green plant production. It was concluded that the D genome plays a crucial role in anther culture response of wheat and that this response is influenced by both the genotype and the duration of cold pre-treatment.


2011 ◽  
Vol 62 (10) ◽  
pp. 813 ◽  
Author(s):  
Sue Broughton

The objective of this study was to improve the production from anther culture of embryos and green plants in Australian spring wheat genotypes by testing new treatments such as n-butanol, as well as other protocol modifications. To date, the use of n-butanol to enhance embryogenesis has only been tested in two European wheat cultivars; this is the first study which demonstrates its application across a range of breeding crosses. A 5-h treatment using 0.1 or 0.2% (v/v) n-butanol following anther pretreatment on a solid mannitol medium significantly improved the production of embryos, green plants and doubled haploids in a range of Australian wheat crosses and varieties. Green plant production increased between 3- and 6-fold in the crosses Yitpi/2*Bumper, Tammarin Rock/2*Bumper and Tammarin Rock/2*Magenta. The addition of calcium (Ca) and macronutrients to the mannitol pretreatment medium also significantly improved the number of embryos and green plants in varieties and crosses, but only when used in combination with n-butanol treatment. A factorial experiment with four varieties and two treatments (n-butanol and Ca/macronutrients) revealed significant interactions between treatments and genotype. In three of the four varieties, the application of n-butanol resulted in significant increases in embryos and green plants with either pretreatment medium although the best results were obtained with Ca and macronutrients in the pretreatment medium, with 200, 193 and 52 green plants per 100 anthers obtained for Bumper, Gladius and Magenta, respectively. In the variety Fortune however, n-butanol treatment did not improve embryo or green plant production unless it was combined with Ca and macronutrients in the pretreatment medium and then there were dramatic improvements; from 0 to 27 green plants per 100 anthers.


2020 ◽  
Vol 26 ◽  
pp. 207-211
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
M. S. Chekalova ◽  
E. A. Golub

Aim. Study the the efficiency of androgenesis in vitro in anther culture of different genotyps winter wheat. Methods. Obtaining of soft winter wheat double haploid lines by anther culture in vitro. The statistical methods. Results The ability to androgenesis in an in vitro anthers culture of 30 soft winter wheat genotypes was tested. The differences in the frequency of callus induction and the ability to regenerate plants in the process of androgenesiss in vitro of winter soft wheat were detected. The range of variation haploproduction activiti (from % of implanted anthers) was broad. The sign of "the formation of callus" was in limited from 0 to 13.2 % and on the sign of "regeneration of green plants" – from 0 to 1.8 %. The 126 green plants-regenerants were received. Conclusions. Genotype-specific of microspores morphogenetic reactions of soft winter wheat in the process of androgenesis in vitro were revealed. It is advisable to use the anthers from the F2 hybrid population plant as a donor material to create an effective technology for producing double haploids of soft winter wheat was shown. Keywords: wheat hybrid, anther culture in vitro, callus, regeneration.


Genetika ◽  
2020 ◽  
Vol 52 (1) ◽  
pp. 335-350
Author(s):  
Osama Kanbar ◽  
Csaba Lantos ◽  
Erzsebet Kiss ◽  
Janos Pauk

The androgenic parameters were investigated by in vitro anther culture (AC) on nine F2-5 breeding combinations of winter wheat (Triticum aestivium L.). Each combination produced embryo-like structures (ELS), green plantlets, albino plantlets, transplanted plantlets and acclimatized plantlets, with respect to the number of anthers in AC. The number of AC-derived ELS was between 11.73 and 52.76 ELS/100 anthers with the mean of 26.22 ELS/100 anthers, out of which the number of regenerated green plantlets varied from 3.20 to 26.40 green plantlets/100 anthers and the mean was 9.76 green plantlets/100 anthers, while the number of transplanted plantlets ranged from 2.16 to 21.77 transplanted plantlets/100 anthers. Furthermore, the number of albinos/100 anthers was mitigated and varied between 0.72 and 6.20 albinos/100 anthers. We also studied the number of green and albino plantlets per 100 ELS. The rate of green plantlets per 100 ELS ranged between 14.81% and 64.01%, with the overall mean 33.59%, while the rate of albinos per 100 ELS ranged from 4.82% to 20.70% with the overall mean 11.93%. In our experiment, the rate of acclimatized plantlets (70.15-91.57%) depended mostly on the combination. This study asserted the importance of AC method in wheat for in vitro production of green plants. Although albinism was found in each combination, it did not hinder the production of green plantlets. The satisfying results were achieved in green plantlets production compared to the previously published data, but further improvement will be needed continuously, experiment by experiment. The generated acclimatized plantlets will be used in the wheat breeding program as doubled haploid (DH) lines.


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