scholarly journals The effect of copper on plant regeneration in barley microspore culture

2017 ◽  
Vol 53 (No. 1) ◽  
pp. 17-22 ◽  
Author(s):  
K. Makowska ◽  
S. Oleszczuk ◽  
J. Zimny
Keyword(s):  
Plant Regeneration ◽  
Copper Sulphate ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Spring Barley ◽  
Culture Method ◽  
Induction Medium ◽  
Isolated Microspore Culture ◽  
Albino Plants ◽  
Effect Of Copper

Isolated microspore culture is an excellent system for the production of doubled haploids in many crops, including barley. In a more traditional barley anther culture method copper sulphate is known to enhance plant regeneration. Here we report that one hundred times higher concentration of copper sulphate in the isolated microspore culture of two spring barley genotypes compared to the standard content in the induction medium resulted in a 34% increase of total plant regeneration. Detailed analysis of plant regeneration showed that additional supplementation of copper sulphate increased not only the regeneration of green plants but also proportionately that of albino plants. Hence, the results from two studied genotypes do not support an assumption that the addition of copper reduces albinism in barley microspore culture.

scholarly journals Production and utilization of doubled haploids in Brassica oleracea vegetables

2011 ◽  
Vol 31 (No. 4) ◽  
pp. 119-123 ◽  
Author(s):  
M. Klíma ◽  
M. Vyvadilová ◽  
V. Kučera
Keyword(s):  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Doubled Haploid Line ◽  
Culture Technique ◽  
F1 Hybrids ◽  
Line Production ◽  
Mature Embryos ◽  
Whole Plants

A possibility to increase the efficiency of plant regeneration from microspore-derived embryos of selected botanical varieties of Brassica oleracea was investigated from 2001 to 2004. More than 400 regenerants of R<sub>1 </sub>generation were derived in kohlrabi, cabbage and cauliflower by means of different modifications of microspore culture technique. Distinct genotype differences in embryogenic responsibility and regenerative ability of microspore embryos to whole plants were detected. The highest frequency of embryogenesis and subsequent regeneration of plants were achieved in cauliflower cultivar Siria F1, kohlrabi line P7 and some experimental F1 hybrids of cauliflower. The best production of embryos was obtained when donor plants were grown in the growth chamber under controlled light and temperature conditions. The regeneration of plantlets was considerably improved by repeated subculture of cotyledonary embryos on media with various combinations of phytohormones and excision of the cotyledons from mature embryos. The percentage of plant regeneration from subcultured embryos in kohlrabi ranged from 11.11 to 63.64%, in cauliflower from 23.53 to 46.19% and in cabbage from 5.88 to 52.00%. The utilization of regenerants for doubled haploid line production is often complicated by male sterility also in plants with the normal diploid chromosome number. &nbsp; &nbsp;

scholarly journals Changes in plastid biogenesis leading to the formation of albino regenerants in barley microspore culture

2020 ◽  
Author(s):  
Monika Gajecka ◽  
Marek Marzec ◽  
Beata Chmielewska ◽  
Janusz Jelonek ◽  
Justyna Zbieszczyk ◽  
...  
Keyword(s):  
Microspore Culture ◽  
Doubled Haploids ◽  
Microspore Embryogenesis ◽  
Rrna Genes ◽  
23S Rrna ◽  
Gene Encoding ◽  
Plastid Differentiation ◽  
Barley Cultivars ◽  
Albino Plants ◽  
Genes Encoding

Abstract Background: Microspore embryogenesis is potentially the most effective method of obtaining doubled haploids (DH), which are utilized in breeding programs to accelerate production of new cultivars. However, the regeneration of albino plants significantly limits the exploitation of androgenesis for DH production in cereals. Despite many efforts, the precise mechanisms leading to development of albino regenerants have not yet been elucidated. The objective of this study was to reveal the genotype-dependent molecular differences in chloroplast differentiation that lead to the formation of green and albino regenerants in microspore culture of barley.Results: We performed a detailed analysis of plastid differentiation at successive stages of androgenesis in two barley cultivars, ‘Jersey’ and ‘Mercada’ that differed in their ability to produce green regenerants. We demonstrated the lack of transition from the NEP-dependent to PEP-dependent transcription in plastids of ‘Mercada’ that produced mostly albino regenerants in microspore culture. The failed NEP-to-PEP transition was associated with the lack of activity of Sig2 gene encoding a sigma factor necessary for transcription of plastid rRNA genes. The impaired PEP activity caused a very low level of 16S and 23S rRNA transcripts, lack of plastid translation machinery and inhibition of photomorphogenesis in regenerating embryos and albino regenerants. Furthermore, the plastids present in differentiating ‘Mercada’ embryos contained a low number of plastome copies whose replication was not always completed. Contrary to ‘Mercada’, ‘Jersey’ that produced 90% green regenerants, showed the high activity of PEP, the highly increased expression of Sig2, plastid rRNA and tRNAGlu transcripts, which indicated the NEP inhibition. The increased expression of GLKs genes encoding transcription factors required for induction of photomorphogenesis was also observed in ‘Jersey’ regenerants. Conclusions: Proplastids present in microspore-derived embryos of albino-producing genotypes did not pass the early checkpoint of their development that are required for induction of further light-dependent differentiation of chloroplasts. The failed activation of plastid-encoded RNA polymerase during differentiation of embryos was the main cause of the genotype-dependent inability to regenerate green plants in barley microspore culture. The better understanding of molecular mechanism underlying formation of albino regenerants may be helpful in overcoming the problem of albinism in cereal androgenesis.

scholarly journals Changes in plastid biogenesis leading to the formation of albino regenerants in barley microspore culture

2020 ◽  
Author(s):  
Monika Gajecka ◽  
Marek Marzec ◽  
Beata Chmielewska ◽  
Janusz Jelonek ◽  
Justyna Zbieszczyk ◽  
...  
Keyword(s):  
Microspore Culture ◽  
Doubled Haploids ◽  
Microspore Embryogenesis ◽  
Rrna Genes ◽  
23S Rrna ◽  
Gene Encoding ◽  
Plastid Differentiation ◽  
Barley Cultivars ◽  
Albino Plants ◽  
Genes Encoding

Abstract Background: Microspore embryogenesis is potentially the most effective method of obtaining doubled haploids (DH) which are utilized in breeding programs to accelerate production of new cultivars. However, the regeneration of albino plants significantly limits the exploitation of androgenesis for DH production in cereals. Despite many efforts, the precise mechanisms leading to development of albino regenerants have not yet been elucidated. The objective of this study was to reveal the genotype-dependent molecular differences in chloroplast differentiation that lead to the formation of green and albino regenerants in microspore culture of barley.Results: We performed a detailed analysis of plastid differentiation at successive stages of androgenesis in two barley cultivars, ‘Jersey’ and ‘Mercada’ that differed in their ability to produce green regenerants. We demonstrated the lack of transition from the NEP-dependent to PEP-dependent transcription in plastids of cv. ‘Mercada’ that produced mostly albino regenerants in microspore culture. The failed NEP-to-PEP transition was associated with the lack of activity of Sig2 gene encoding a sigma factor necessary for transcription of plastid rRNA genes. A very low level of 16S and 23S rRNA transcripts and impaired plastid translation machinery resulted in the inhibition of photomorphogenesis in regenerating embryos and albino regenerants. Furthermore, the plastids present in differentiating ‘Mercada’ embryos contained a low number of plastome copies whose replication was not always completed. Contrary to ‘Mercada’, cv. ‘Jersey’ that produced 90% green regenerants, showed the high activity of PEP polymerase, the highly increased expression of Sig2, plastid rRNAs and tRNAGlu, which indicated the NEP inhibition. The increased expression of GLKs genes encoding transcription factors required for induction of photomorphogenesis was also observed in ‘Jersey’ regenerants. Conclusions: Proplastids present in microspore-derived embryos of albino-producing genotypes did not pass the early checkpoint of their development that are required for induction of further light-dependent differentiation of chloroplasts. The failed activation of plastid-encoded RNA polymerase during differentiation of embryos was associated with the genotype-dependent inability to regenerate green plants in barley microspore culture. The better understanding of molecular mechanism underlying formation of albino regenerants may be helpful in overcoming the problem of albinism in cereal androgenesis.

scholarly journals Androgenesis of Red Cabbage in Isolated Microspore Culture In Vitro

Plants ◽  
2021 ◽  
Vol 10 (9) ◽  
pp. 1950
Author(s):  
Anna Mineykina ◽  
Ludmila Bondareva ◽  
Alexey Soldatenko ◽  
Elena Domblides
Keyword(s):  
Culture Medium ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Antioxidant Properties ◽  
F1 Hybrids ◽  
Anthocyanin Content ◽  
Isolated Microspore Culture ◽  
Culture In Vitro ◽  
Red Cabbage

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.

scholarly journals Effects of Genotype and Culture Conditions on Microspore Embryogenesis and Plant Regeneration in Brassica Rapa ssp. Rapa L.

Plants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 278 ◽  
Author(s):  
Daria Shumilina ◽  
Dmitry Kornyukhin ◽  
Elena Domblides ◽  
Alexey Soldatenko ◽  
Anna Artemyeva
Keyword(s):  
Microspore Culture ◽  
Doubled Haploids ◽  
Genetic Disorder ◽  
Cold Treatment ◽  
Microspore Embryogenesis ◽  
Culture Conditions ◽  
Secondary Embryogenesis ◽  
Isolated Microspore Culture ◽  
Pure Lines

Turnip is a biennial crop and, consequently, the creation of pure lines for breeding is a time-consuming process. The production of pure turnip lines using doubled haploids produced in isolated microspore culture has not been sufficiently developed. The aim of the present work was to determine some key factors inducing embryogenesis in the isolated microspore culture of turnip, as well as investigating the manners of embryo development. It was shown that the acidity of the medium is an important factor in embryo production; different optimal pH levels ranging from 6.2 to 6.6 corresponded to individual genotypes. Such factors as the cold treatment of buds and the addition of activated charcoal to the nutrient medium increased the responsiveness of all genotypes studied. The turnip variety ‘Ronde witte roodkop herfst’ demonstrated a genetic disorder in the development of microspores; namely, non-separation of some microspores from tetrads. In the in vitro culture, each of the daughter microspores developed on its own. This indicates the dependence of the possibility of embryogenesis in the turnip microspore culture on the genotype. Results suggest that the initiation of secondary embryogenesis in primary embryos leads to an increase in the proportion of doubled haploid plants.

Production of doubled haploids in durum wheat (Triticum turgidum L.) through isolated microspore culture

2005 ◽  
Vol 25 (4) ◽  
pp. 257-264 ◽  
Author(s):  
L. Cistué ◽  
M. Soriano ◽  
A. M. Castillo ◽  
M. P. Vallés ◽  
J. M. Sanz ◽  
...  
Keyword(s):  
Durum Wheat ◽  
Triticum Turgidum ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Isolated Microspore Culture

Ploidy of Regenerated Broccoli Derived from Microspore Culture Versus Anther Culture

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 533g-534
Author(s):  
Min Wang ◽  
Mark W. Farnham
Keyword(s):  
Anther Culture ◽  
Doubled Haploid ◽  
Direct Method ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Chromosome Complement ◽  
Culture Method ◽  
Lower Percentage ◽  
Ploidy Levels ◽  
Culture Techniques

Anther and microspore culture are commonly utilized to produce doubled-haploid (diploid), homozygous lines in broccoli (Brassica oleracea L. Italica Group). It is well-documented that doubled-haploid regenerants are produced by means of polyploidization during anther culture. However, polyploidization may not occur at all, or it may involve a tripling or quadrupling of the chromosome complement. As a consequence, regenerated populations from anther culture contain diploids, but also haploids, triploids, and tetraploids. Microspore culture represents a simpler and more direct method for producing doubled-haploids. Although a similar mix of ploidy types is likely to be observed among regenerants derived from microspore culture, the actual ploidy levels of such regenerants have not been documented for broccoli. Thus, the objectives of this study were to compare ploidy levels of regenerants developed using both anther and microspore culture in broccoli, and to examine phenotypic variation in ploidy makeup of populations developed from both anther and microspore culture using different F1 hybrids. Broccoli regenerants were derived simultaneously from both anther and microspore cultures using the same four F1 hybrids, including Everest, Patriot, Greenbelt and Major. Ploidy level was determined by flow cytometry. A majority of regenerants derived from both anther and microspore culture, were determined to be diploids or tetraploids. Significant differences in ploidy makeup of populations were observed among hybrid varieties for both culture techniques. Regardless of the culture method used, `Everest' produced a greater percentage of diploids and a lower percentage of tetraploids than `Patriot' did. Haploids were observed more frequently from microspore culture than from anther culture when `Everest' and `Major' served as parents.

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