scholarly journals Effect of sowing dates of donor plants and concentration of 2,4-D on the purity of productive anther formation of common barley (Hordeum vulgare L.) in anther culture in vitro.

Author(s):  
K. I. Popova ◽  
J. S. Skryabin ◽  
P. A. Lyakh ◽  
N. V. Petrash

Creating dihaploid lines of agricultural plants is a labour-intensive but essential step in variety production in modern plant breeding. This stage allows significantly accelerate the process of creating new varieties of common barley and other crops. Barley digaploids are produced mainly by anther culture and microspore culture. The authors preferred anther culture in vitro. In the present study, the influence of climatic factors in the cultivation of donor plants on the yield of productive anthers at different sowing dates was established. The authors also identified the more stable culti- vars with a high anther production regardless of sowing date (Signal, Laureate and Eifel). Varieties showed the highest number of embryo-like structures formation at the first and third sowing dates (Zu Suren, Zu Zaza); and sorts with a high rate of productive anther formation at the second sowing date (Acha, Exploer) were identified. Different concentrations of 2,4-D in N6 medium on the frequency of embryogenesis and yield of productive anthers were studied. As a result of this study, the authors found that different concentrations of 2,4-D (1 mg/l and two mg/l) had no significant effect on the for- mation frequency of productive anthers in all the varieties studied. When the embryogenesis capacity of the cultivars was reviewed, all the samples were found to be positive in anther culture. However, the array Zu Suren had a significantly lower effective anthers yield than the samples Signal and Acha. As a result of correlation analysis, the authors found a close relationship between the length of the ear tube of donor plants and the frequency of formation of productive anthers (r = -0.69). A close relationship with the development of optimal microspore phase for the induction of androgenesis in anthers extracted from the ear tube with an average length of 6 cm was determined. This information can significantly speed up the selection of donor plants, but it is recommended to confirm the stage of microspore development microscopically for each new cultivar used.

2019 ◽  
Vol 25 ◽  
pp. 178-183
Author(s):  
O. V. Bilinska ◽  
P. G. Dulnyev

Aim. Evaluation of innovative methodological approaches elaborated on model genotypes for usability to increase frequencies of morphogenic structure induction and plant regeneration in anther culture in vitro in spring barley diverse material. Methods. Spikes isolated from F1 and F2 hybrids of four crosses were pretreated using an improved method (4ºC, 28 days), and anthers were inoculated onto nutrient media containing chemically modified starches D5-M and D5a-1 instead of agar. In control cut tillers were emerged in water and pretreated at 4ºC for 5 days. Anthers were cultivated on agar solidified medium. Results. Positive effects of the improved method of cold pretreatment and cultivation of anthers on media solidified with starches were confirmed. The advantage of new gelling agent D5a-1 was proved. Particularly, its usage resulted in a three-fold increase in the frequency of green plant regeneration. Conclusions. In order to increase spring barley androgenic haploid yield, combination of prolonged cold pretreatment with anther cultivation on media solidified with chemically modified starches instead of agar in an integrated technological process is reasonable. Keywords: Hordeum vulgare L., anther culture in vitro, cold pretreatment, starch, agar, embryo formation, plant regeneration.


Plants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1950
Author(s):  
Anna Mineykina ◽  
Ludmila Bondareva ◽  
Alexey Soldatenko ◽  
Elena Domblides

Red cabbage belongs to the economically important group of vegetable crops of the Brassicaceae family. A unique feature of this vegetable crop that distinguishes it from other members of the family is its unique biochemical composition characterized by high anthocyanin content, which gives it antioxidant properties. The production mainly uses F1 hybrids, which require constant parental lines, requiring 6–7 generations of inbreeding. Culture of isolated microspores in vitro is currently one of the promising methods for the accelerated production of pure lines with 100% homozygosity. The aim of this study is to investigate the factors and select optimal parameters for successful induction of red cabbage embryogenesis in isolated microspore culture in vitro and subsequent regeneration of DH plants. As a result of research, for the first time, it was possible to carry out the full cycle of obtaining DH plants of red cabbage from the induction of embryogenesis to their inclusion in the breeding process. The size of buds containing predominantly microspores at the late vacuolated stage and pollen at the early bi-cellular stage has to be selected individually for each genotype, because the embryoid yield will be determined by the interaction of these two factors. In the six samples studied, the maximum embryoid yield was obtained from buds 4.1–4.4 mm and 4.5–5.0 mm long, depending on the genotype. Cultivation of microspores was carried out on liquid NLN culture medium with 13% sucrose. The maximum number of embryoids (173.5 ± 7.5 pcs./Petri dish) was obtained on culture medium with pH 5.8 and heat shock at 32 °C for 48 h. Successful embryoid development and plant regeneration by direct germination from shoot apical meristem were achieved on MS culture medium with 2% sucrose and 0.7% agar, supplemented with 6-benzylaminopurine at a concentration of 1 mg/L. Analysis of the obtained regenerated plants, which successfully passed the stage of adaptation to ex vitro conditions by flow cytometry, showed that most of them were doubled haploids (up to 90.9%). A low number of seeds produced by self-fertilization in DH plants was observed.


Author(s):  
M. V. Ilyushko ◽  
M. V. Romashova

Morphological variability of haploid plants and doubled haploids of rice obtained on one callus line in anther culture in vitro was studied. The work was carried out on rice plants Oryza sativa L. subspecies japonica Kato varietу Cascade. Regenerant plants of one callus line obtained from one rice anther (four in total) were divided into two or three groups of 20-30 plants, depending on the sample size in order of their differentiation on callus and transplantation on the rooting medium. Two callus lines (15.1 and 18.1) formed half of the haploids, half of the doubled haploids, and two other callus lines (5.1 and 7.2) numerous haploids. On callus lines with numerous haploids (5.1 and 7.2), as the regenant number increases, the size of plants decreases (plant height, number of flowers on the main panicle, number of panicles). On the lines 15.1 and 18.1 between groups of haploids and between groups of doubled haploids statistically significant differences absent. In breeding purposes for the induced doubling of the number of chromosomes in haploid regenerants with antitubulin substances such as colchicine, it is advisable to use plants that form on callus among the first. Between haploids of four callus lines and doubled haploids of two callus lines, statistically significant differences (at p=0.001) were revealed using the Hotelling's T2-criterion, calculated for the whole complex of biometric features. Haploids of different lines differed in three or four of them, doubled haploids on three of the five signs (length of panicle, productive bushiness and plant height). Varieties of interest to breeders may be improved by anther culture in vitro.


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 14 (2) ◽  
pp. 178-180
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
D. V. Shpak ◽  
A. O. Dobrova ◽  
S. O. Ignatova

Aims. To study the effect of chemically modified starch D–5aM in the culture medium on the efficiency of androgenesis in vitro in anther culture of rice. Methods. Obtaining of rice double haploid lines by anther culture in vitro. The statistical methods. Results The influence different variants of gellatyne source in culture medium on the processes of induction and regeneration in anther culture of rice were studied. The 119 green plants-regenerants were received. Conclusions. The negative effect on the formation of green regenerants using a gel-forming components of the chemically modified starch D–5aM was shown. Keywords: rice, anther culture in vitro, callus, regeneration, chemically modified starch.


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.


Sign in / Sign up

Export Citation Format

Share Document