Effect of Nutrient Media Containing Natural and Chemically Modified Starches on Haploid Production in Spring Barley Anther Culture in vitro

2015 ◽  
pp. 239-256
Keyword(s):  
Anther Culture ◽  
Spring Barley ◽  
Nutrient Media ◽  
Culture In Vitro ◽  
Chemically Modified ◽  
Haploid Production ◽  
Modified Starches

scholarly journals Efficiency of spring barley haploid production in anther culture in vitro: comparison of basic and improved technologies

2019 ◽  
Vol 25 ◽  
pp. 178-183
Author(s):  
O. V. Bilinska ◽  
P. G. Dulnyev
Keyword(s):  
Plant Regeneration ◽  
Anther Culture ◽  
Spring Barley ◽  
Gelling Agent ◽  
Improved Method ◽  
Cold Pretreatment ◽  
Culture In Vitro ◽  
Chemically Modified ◽  
Modified Starches

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.

scholarly journals Grain barley starch as a gelling agent of nutrient media for cultivation of anthers and isolated embryos of H. vulgare L.

2020 ◽  
Vol 26 ◽  
pp. 158-163
Author(s):  
O. V. Bilynska
Keyword(s):  
Embryo Culture ◽  
Spring Barley ◽  
Gelling Agent ◽  
Modified Starch ◽  
Normal Type ◽  
Nutrient Media ◽  
Culture In Vitro ◽  
Chemically Modified ◽  
Pea Starch

Aim. Determination the possibility of agar replacement with normal and waxy type starches produced from barley seeds in the nutrient media for spring barley haploid production. Methods. Anthers of spring barley line DH00-126 were inoculated on inductive media containing N6 macro-, MS micronutrients, organic supplements, maltose (9.0 %) and differed in solidifying agents (agar, starch from barley seeds or chemically modified starch D5a-1). Trophic capacities of barley starch were determined both in anther and embryo culture of barley. Results. It was determined that barley starch of waxy type was not suitable for solidifying of nutrient medium. Use of normal type barley starch (6.5 %) resulted in sufficient decrease in the frequency of green plant regeneration. At the same time, this modification had no effect on the number of morphogenic anthers. Barley starch was inferior to pea starch in the ability to support seedling growth in embryo culture in vitro. Conclusions. Barley starch of normal type has worse trophic capacities in comparison to pea starch in embryo culture. It was appeared barley starch to be a less suitable gelling agent of medium for spring barley haploid production in anther culture in vitro than agar and chemically modified starch D5a-1. Keywords: Hordeum vulgare L., anther culture in vitro, embryo culture in vitro, agar, barley starch.

scholarly journals Effect of trophic, osmotically active and solidifying components of nutrient medium on the direct embryognesis in spring barley anther culture in vitro

2018 ◽  
Vol 22 ◽  
pp. 210-215
Author(s):  
E. V. Belinskaya ◽  
P. G. Dulnyev
Keyword(s):  
Plant Regeneration ◽  
Anther Culture ◽  
Spring Barley ◽  
Gelling Agent ◽  
Modified Starch ◽  
Hordeum Vulgare L ◽  
Culture In Vitro ◽  
Chemically Modified ◽  
Determine Mechanism

Aim. Ambryoidogenesis, or somatic embryogenesis, is known to be the most efficient mode of plant regeneration in plant cell, tissue and organ culture. The investigation was aimed to elucidate effects of trophic and osmogenic components of inductive medium on the frequency of direct embryoidogenesis in spring barley anther culture in vitro and to determine mechanism of morphogenesis improvement coursed by chemically modified starch D-5aM used as a gelling agent instead of agar. Methods. Anthers of DH-line with a high androgenetic capacity were inoculated on inductive media containing N6 macro-, MS micronutrients, organic supplements, maltose or mannitol (0.3 M) and solidified with agar or chemically modified starch. Results. A positive effect of combination of high maltose content and chemically modified starch on the induction and regeneration processes in spring barley anther culture in vitro was confirmed. It was also shown that mannitol didn’t keep any growth or development processes going in barley anther and embryo culture, but at the same time this substance had no toxic effect. Conclusions. In order to achieve a high frequency of induction in spring barley anther culture, it is necessary to use medium containing maltose – a low weight component both with trophic and with osmotic activity. Keywords: Hordeum vulgare L., anther culture in vitro, mannitol, maltose, embryo formation, plant regeneration.

scholarly journals Effect of medium gelatinized component on the effectiveness of androgenesis in vitro Oryza sativa L.

2016 ◽  
Vol 14 (2) ◽  
pp. 178-180
Author(s):  
I. S. Zambriborshch ◽  
O. L. Shestopal ◽  
D. V. Shpak ◽  
A. O. Dobrova ◽  
S. O. Ignatova
Keyword(s):  
Anther Culture ◽  
Culture Medium ◽  
Oryza Sativa L ◽  
Modified Starch ◽  
Culture In Vitro ◽  
Chemically Modified ◽  
Androgenesis In Vitro ◽  
Negative Effect ◽  
Callus Regeneration

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.

scholarly journals Effect of low molecular weigt osmogenic substanses on the induction of embryogenesis in spring barley anther culture in vitro

1970 ◽  
pp. 164-167
Author(s):  
O. V. Bilynska
Keyword(s):  
Plant Regeneration ◽  
Osmotic Pressure ◽  
Anther Culture ◽  
Spring Barley ◽  
Hordeum Vulgare L ◽  
Culture In Vitro ◽  
Negative Effect ◽  
Embryoid Formation ◽  
The Impact

Aim. Osmotic pressure of nutrient medium is known to be an important factor for morphogenesis in plant cell, tissue and organ culture in vitro. The investigations was aimed to elucidate the effect of maltose concentration and to evaluate the impact of mannitol add on callus, embryoid formation and plant regeneration in spring barley anther culture in vitro. Methods. Anthers of DH-line with a high androgenetic capacity were inoculated on inductive media contained N6 macro-, MS micronutrients, organic supplements, maltose (0−9.0 %) and 0.1 M mannitol. Results. A decrease in maltose concentration from 9 to 6 % had a strong negative effect on these processes. At the same time, addition of 0.1 M mannitol to medium containing 6 % maltose promoted sufficiently increase the efficiency of embryoid formation and plant regeneration. Conclusions. Medium osmotic pressure mainly determined by a high maltose concentration played an important role in spring barley haploid production. Decrease in maltose content could be compensated by addition of mannitol, which is osmotic substance with low metabolic activity. Keywords: Hordeum vulgare L., anther culture in vitro, mannitol, embryo formation, plant regeneration.

Some influence of the hormonal components in nutrient medium onthe induction of novel structures when obtaining rapeseed haploids through in vitro culture

2018 ◽  
pp. 30-37
Keyword(s):  
Anther Culture ◽  
Hormonal Regulation ◽  
Control Plant ◽  
Pollen Grains ◽  
Control Treatment ◽  
Basic Medium ◽  
Nutrient Media ◽  
Oilseed Crops ◽  
Wide Range

Growth regulators, phytohormones, both natural and artificial, are the main means to control plant ontogenesis. They are involved in regulating the processes of cell differentiation and cell divisions, the formation of tissues and organs, the changes in the rate of growth and development, the duration of the certain stages of ontogenesis. The main classes of phytohormones used in plant biotechnology, in particular, in the induction of haploid structures, are auxins and cytokinins. The mechanism of action of phytohormones on a cell is rather complicated and may have a different character. Understanding the characteristics of the action of phytohormones is complicated by the fact that the system of hormonal regulation of plant life is multicomponent. This is manifested in the fact that the same physiological process is most often influenced not by one, but by several phytohormones, covering a wide range of aspects of cell metabolism. In connection with the foregoing, the purpose of our work was to test a set of nutrient media with different basic composition and different proportions of phytohormones to determine the patterns of their influence on the processes of haploid structure induction in rape anther culture using accessions, developed at the Institute of Oilseed Crops NAAS. The material used was two accessions of winter rapeseed (No. 1 and No. 2) and one sample of spring rapeseed, provided by the Rapeseed Breeding laboratory of the Institute of Oilseed Crops. Incised inflorescences were kept against the background of low temperature of 6–8 ° C for several days, and then, under aseptic conditions, anthers with unripe pollen grains were isolated and planted on nutrient media differing in both basic mineral composition and content of phytohormones. MS (Murashige & Skoog 1962) and B5 (Gamborg et al 1968) media were used as basic media. Phytohormones were added to the basic media in various combinations – BA, 2,4-D, NAA at the concentrations of 0.1-0.6 mg/l. In each treatment up to 300 anthers were cultivated. Differences between treatments were evaluated using standard t-test. Studies have shown that in the anther culture of rapeseed on the tested nutrient media, morphogenic structures of different types (embryoids and callus) were originated. Synthetic auxin 2,4-D, regardless of the composition of the basic medium, caused the formation of structures of both types, though with a low frequency. Phytohormone BA of the cytokinin type had a similar effect. In this case, the frequency of structures was slightly higher, and the developed structures were represented mainly by embryoids. The joint action of cytokinin and auxin was the most favorable for the initiation of morphogenic structures. Such combination of phytohormones caused the formation of these structures with a frequency of 24.5-14.7% in the studied genotypes of winter rape. A similar effect of phytohormones on the induction and development of morphogenic structures was also observed in spring rape. In this case, a single basic MS medium was used. The experiment included treatments where phytohormones were absent (control), as well as various combinations of auxin and cytokinin. In the control treatment, the formation of new structures was not noted. In treatments with phytohormones, in addition to the medium with the combination of auxin and cytokinin, the medium in which only cytokinin was present was also rather effective. The treatment in which the action of auxin 2,4-D was combined with the action of another auxin, NAA, turned out to be practically ineffective. Thus, it was found that for the induction of morphogenic structures from microspores in rape anther culture of the tested genotypes, the combination of cytokinin with auxin, or the use of only single cytokinin BA without other phytohormones, had the most positive effect.

scholarly journals Variability of rice haploids obtained in anther culture in vitro

2019 ◽  
pp. 11-14
Author(s):  
M. V. Ilyushko ◽  
M. V. Romashova
Keyword(s):  
Anther Culture ◽  
Plant Height ◽  
Oryza Sativa L ◽  
Doubled Haploids ◽  
Morphological Variability ◽  
Callus Line ◽  
Culture In Vitro ◽  
Panicle Number ◽  
Callus Lines

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.

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