Study of androgenesis and spontaneous chromosome doubling in barley ( Hordeum vulgare L.) genotypes using isolated microspore culture

2009 ◽  
Vol 57 (2) ◽  
pp. 155-164 ◽  
Author(s):  
D. Kahrizi ◽  
R. Mohammadi
Keyword(s):  
Chromosome Doubling ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Negative Relationship ◽  
Microspore Embryogenesis ◽  
Culture Technique ◽  
Hordeum Vulgare L ◽  
Spontaneous Chromosome ◽  
Barley Genotypes

This research aimed to study the androgenesis and spontaneous chromosome doubling of five barley genotypes using an isolated in vitro microspore culture technique, involving a completely randomized design (CRD) with three replications. Statistical analysis of embryogenesis and cytogenetic results showed that genotype had a significant effect on haploid embryogenesis (P<0.01) and on spontaneous chromosome doubling (P<0.05). The genotype Igri was found to have the highest potential to produce haploid embryos (1577 embryos from 100 anthers), followed by the genotypes Boyer/Rojo, Afzal/Turkman/Kavir, Ashar/Hebo and Agrigashar/Matico with 369, 304, 278 and 150 embryos from 100 anthers, respectively. The highest percentage of spontaneous chromosome doubling (76%) was observed for the genotype which had the lowest embryogenesis (Agrigashar/Matico) and the lowest (65%) for the genotype with the highest androgenic capacity (Igri). Microspore embryogenesis also showed comparatively higher genotypic (109.2) and phenotypic (109.5) coefficients of variation, heritability (99.62) and genetic advance (1206.77), indicating the pre-dominance of additive gene action in the control of this character in the material studied. Estimates of genetic parameters (PCV, GCV and heritability) for microspore embryogenesis were higher than for spontaneous doubled haploids. These results indicated that selection for androgenic capacity would be more effective than for spontaneous doubled haploids. The findings showed a negative relationship (r= −0.68) between embryogenesis and spontaneous chromosome doubling in the barley genotypes studied. All the large embryos used had high regenerability and good plantlet formation.

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.

scholarly journals Spontaneous chromosome doubling results from nuclear fusion during in vitro maize induced microspore embryogenesis

Chromosoma ◽  
2004 ◽  
Vol 112 (7) ◽  
pp. 342-349 ◽  
Author(s):  
P. Testillano ◽  
S. Georgiev ◽  
H. L. Mogensen ◽  
M. J. Coronado ◽  
C. Dumas ◽  
...  
Keyword(s):  
Chromosome Doubling ◽  
Nuclear Fusion ◽  
Microspore Embryogenesis ◽  
Spontaneous Chromosome

scholarly journals Microspore embryogenesis in vitro: the role of stresses

2019 ◽  
Vol 23 (1) ◽  
pp. 86-94 ◽  
Author(s):  
T. I. Djatchouk ◽  
O. V. Khomyakova ◽  
V. N. Akinina ◽  
I. A. Kibkalo ◽  
A. V. Pominov
Keyword(s):  
Microspore Culture ◽  
Doubled Haploids ◽  
Embryo Sac ◽  
Microspore Embryogenesis ◽  
Gametic Embryogenesis ◽  
Chemical Pretreatments ◽  
Dh Lines

Gametic embryogenesis is one form of totipotency of plant cells, in which either male or female gametes are induced to form embryoids (sporophytes). Regeneration of haploid plants from embryoids and subsequent chromosome duplication result in doubled haploids and DH-lines. The production of haploids and doubled haploids (DHs) through gametic embryogenesis allows a single-stage development of complete homozygous lines from heterozygous plants. The development of effective haploid protocols to produce homozygous plants has a significant impact on plant breeding, shorting the time and costs required to establish new cultivars. There are several available methods to obtain haploids and DHs-lines, of which anther or isolated microspore culture in vitro are the most effective. Microspore embryogenesis is more commonly applied. This is in part because more male gametophytes are contained in a single anther compared to the single female gametophyte per embryo sac. Microspore embryogenesis is regarded as one of the most striking examples of plant cell totipotency. The switch of cultured microspores from gametophytic to sporophytic mode of development has been induced by stress treatments of various kinds applied to donor plants, inflorescences, buds, anthers or isolated microspores both in vivo and in  vitro. Physical or chemical pretreatments (cold and heat shock, sugar starvation, colchicine, n-butanol, gametocydes) act as a trigger for inducing the sporophytic pathway, preventing the gametophytic pathway development of microspore. The recent investigations have revealed that cold pretreatment during microspore reprogramming acts rather as an anti-stress factor alleviating the real stress caused by nutrient starvation of anthers or microspores isolated from donor plants. Under stress pretreatment a vacuolated and polarized microspore transformed into a depolarized and dedifferentiated cell, which is an obligatory condition for reprogramming their development. We summarize data concerning the role of various stresses in the induction of microspore embryogenesis and possible mechanisms of their action at cellular and molecular levels. Identification of new stresses allows creating efficient protocols of doubled haploid production for end-user application in the breeding of many important crops.

scholarly journals The Influence of pH on Microspore Embryogenesis of White Cabbage (Brassica oleracea L.)

2013 ◽  
Vol 5 (4) ◽  
pp. 485-489 ◽  
Author(s):  
Tina Oana CRISTEA
Keyword(s):  
Brassica Oleracea ◽  
Microspore Culture ◽  
Microspore Embryogenesis ◽  
Critical Factor ◽  
Cellular Level ◽  
Strong Base ◽  
White Cabbage ◽  
In Vitro Microspore Culture ◽  
Influence Of Ph

In vitro microspore culture is one of the top techniques utilised now-a-days for the obtaining of double haploid plants in many plant species, including Brassica. The pH of the medium is a critical factor for the success of In vitro microspore culture as it influences the invertase enzyme activity, translated at cellular level through an acceleration or reduction of sucrose cleavage. The results published until now shows rather contradictory findings, as the response of microspores have been proved to be highly depending on genotypes, most of them being focused on Brassica napus. Thus, in the present study, the effect of different NLN liquid medium pH, ranging between 5.0 to 7.0 were tested in order to establish the most suitable pH for the expression of embryogenic competences of microspores cultivated on medium In vitro and ultimately for the obtaining of microspore-derived embryos. Among the 11 values of pH tested, the best results were obtained on variants with pH 5.8 and 6.0, both in what concern the maintaining of microspores viability and the number of microspore-derived embryos. The findings of the present study provide a strong base for the establishment of an efficient protocol for the In vitro culture of microspore at Brassica oleracea L. genotypes with Romanian origin.

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;

Performance of isolated microspore-derived doubled haploids of wheat (Triticum aestivum L.)

1997 ◽  
Vol 77 (4) ◽  
pp. 549-554 ◽  
Author(s):  
Tianci Hu ◽  
Ken J. Kasha
Keyword(s):  
In Vitro Selection ◽  
Nuclear Dna ◽  
Chromosome Doubling ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Spontaneous Mutation ◽  
Nuclear Dna Content ◽  
Root Tip ◽  
Ploidy Levels ◽  
Dh Lines

Ploidy level, genetic stability and field performance of isolated microspore-derived wheat plants were evaluated. The ploidy levels of isolated microspore-derived wheat plants from cv. Chris and reciprocal crosses of Chris × Sinton were determined by two methods, namely chromosome counts of root tip cells and flow cytometric measurement of nuclear DNA content from leaves. Both methods gave similar results with the frequencies of spontaneous chromosome doubling and completely fertile plants among microspore-derived H0 plants of wheat being about 80% and 75%, respectively, based on two H0 populations. Only 1.7% were aneuploid and 16% were haploid. Spontaneous mutation frequencies were low with 1 of 124 Chris DH plants having a recessive mutant for lemma awns while three were grass-like plants in the H0 generation. The field-measured traits of microspore-derived DH lines were similar to the check by the third generation. Ninety-two percent and 70% of Chris DH lines had 1000-kernel weights and yields similar to the check, respectively. These results indicate that microspore-derived DH lines should have good potential for applications in plant breeding, in vitro selection, plant transformation and genetic studies. Key words: Wheat, microspore, culture, haploidy, performance, stability

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 An efficient regeneration system through in vitro somatic embryogenesis of barley (Hordeum vulgare L.)

2019 ◽  
Vol 27 ◽  
pp. 89-99
Author(s):  
M Haque ◽  
SMS Islam
Keyword(s):  
Plant Regeneration ◽  
Callus Induction ◽  
Casein Hydrolysate ◽  
Ms Medium ◽  
Regeneration System ◽  
Embryogenic Calli ◽  
Hordeum Vulgare L ◽  
The Media ◽  
Barley Genotypes

This study was carried out to improve an efficient protocol for in vitro callus induction and plant regeneration using Bangladeshi barley genotypes collected from BARI, Gazipur, Bangladesh. After sterilization embryos were separated carefully from mature seeds of six barley genotypes (BB-1, BB-2, BB-3, BB-4, BB-5 and BB-6) and cultured them in MS medium supplemented with various concentration and combination of PGRs for callus induction and regeneration. Out of six genotypes BB-6 showed highest (38.17%) callus induction in MS + 4.0 mg/l 2,4-D + 200 mg/l L-proline + 300 mg/l casein hydrolysate; whereas, BB-4 and BB-5 showed no callus induction in the same medium. For plant regeneration from embryogenic calli the same genotype (BB-6) also performed the best results (19.25%) in MS medium supplemented with 1.5 mg/l BAP + 30 g/l sucrose. Analysis of variance (ANOVA) showed highly significant differences among the media and the genotypes. J. bio-sci. 27: 89-99, 2019

Sign in / Sign up

Export Citation Format

Share Document