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;

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 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 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 Doubled Haploids in Eggplant

Biology ◽  
2021 ◽  
Vol 10 (7) ◽  
pp. 685
Author(s):  
Ricardo Mir ◽  
Antonio Calabuig-Serna ◽  
Jose M. Seguí-Simarro
Keyword(s):  
Anther Culture ◽  
Germ Line ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
F1 Hybrids ◽  
Breeding Programs ◽  
Isolated Microspores ◽  
Somatic Tissues ◽  
Pure Lines ◽  
Made In

Eggplant is a solanaceous crop cultivated worldwide for its edible fruit. Eggplant breeding programs are mainly aimed to the generation of F1 hybrids by crossing two highly homozygous, pure lines, which are traditionally obtained upon several self crossing generations, which is an expensive and time consuming process. Alternatively, fully homozygous, doubled haploid (DH) individuals can be induced from haploid cells of the germ line in a single generation. Several attempts have been made to develop protocols to produce eggplant DHs principally using anther culture and isolated microspore culture. Eggplant could be considered a moderately recalcitrant species in terms of ability for DH production. Anther culture stands nowadays as the most valuable technology to obtain eggplant DHs. However, the theoretical possibility of having plants regenerated from somatic tissues of the anther walls cannot be ruled out. For this reason, the use of isolated microspores is recommended when possible. This approach still has room for improvement, but it is largely genotype-dependent. In this review, we compile the most relevant advances made in DH production in eggplant, their application to breeding programs, and the future perspectives for the development of other, less genotype-dependent, DH technologies.

scholarly journals Effects of Broccoli Donor Genotype on Incidence of Diploids in Populations Regenerated from Anther Culture

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 612g-613
Author(s):  
Mark W. Farnham
Keyword(s):  
Flow Cytometry ◽  
Plant Regeneration ◽  
Brassica Oleracea ◽  
Anther Culture ◽  
High Frequency ◽  
Microspore Culture ◽  
Chromosome Complement ◽  
Ploidy Levels ◽  
Previous Cycle ◽  
Donor Genotype

Broccoli (Brassica oleracea L. Italica group) breeders are increasingly using anther or microspore culture to produce dihaploid (diploid), homozygous lines for use in making hybrids. During the process of anther culture and subsequent plant regeneration, wherein embryos develop from microspores and plants are regenerated from the embryos, polyploidization occurs and diploid regenerants can result. However, polyploidization may not occur at all, or it may involve a tripling or quadrupling of the chromosome complement, instead of a doubling. Thus, populations may contain haploids, triploids, or tetraploids, in addition to diploids. In two cycles (1994-95 and 1995-96) of anther culture, regenerated populations from different broccoli hybrid sources were evaluated using flow cytometry to facilitate efficient identification of diploids vs. haploids, tetraploids, or others and to determine if anther donor genotype has an effect on the frequency of different ploidy levels among regenerants. In the first cycle, five broccoli hybrids had anther-derived populations in which ≈33% were haploid, 55% diploid, 37% tetraploid, and 5% aneuploid or abherent types. The hybrid, `Marathon', was different; it's regenerants were 78% diploid and only 15% tetraploid. In the second cycle, anther-derived populations had a significantly different makeup with a most hybrids giving 30% to 40% diploids and 50% to 60% tetraploids. However, consistent with the previous cycle, `Marathon' gave significantly more diploids (68%) and fewer tetraploids (25%) than other hybrids. These results indicate that anther donor genotype affects ploidy frequency among regenerants. Genotypes producing a high frequency (>60%) of diploids may be relatively uncommon.

scholarly journals Comparison of methods for obtaining doubled haploids of carrot

2017 ◽  
Vol 86 (2) ◽  
Author(s):  
Waldemar Kiszczak ◽  
Urszula Kowalska ◽  
Agata Kapuścińska ◽  
Maria Burian ◽  
Krystyna Górecka
Keyword(s):  
Plant Regeneration ◽  
Doubled Haploid ◽  
Aspartate Aminotransferase ◽  
Doubled Haploids ◽  
The Other ◽  
Plant Production ◽  
Comparison Of Methods ◽  
Anther Cultures ◽  
Induction Stage ◽  
Main Factor

Doubled haploid lines of carrot can be obtained through androgenesis in anther cultures and in isolated microspore cultures. The two methods were compared using three carrot cultivars (‘Kazan F1’, ‘Feria F1’, and ‘Narbonne F1’) at the androgenesis induction stage, during plant regeneration from embryos, and during acclimatization of androgenetic plants as well as their characterization. It was found that cultivar was the main factor affecting the efficiency at each stage of plant production in both anther and isolated microspore cultures. The efficiency of androgenesis in anther cultures of ‘Feria F1’ was considerably higher in comparison with isolated microspore cultures, and more plants were obtained from the embryos of androgenesis-cultured plants. In ‘Kazan F1’ and ‘Narbonne F1’, more acclimatized androgenetic plants were produced from anther cultures. Ploidy assessment of acclimatized plants of ‘Narbonne F1’ showed that the majority of the plants in the population derived from anther cultures had a doubled chromosome (DH) set. On the other hand, the majority of plants obtained from isolated microspore cultures were haploids. When assessing homozygosity, it was found among plants obtained in anther cultures that the percentage of homozygotes for phosphoglucose isomerase (PGI) and aspartate aminotransferase (AAT) depended on the cultivar. In contrast, the majority of plants derived from isolated microspore cultures were homozygous regardless of cultivar.

scholarly journals Ação de extratos vegetais e fitoreguladores na organogênese de Begonia rex

1998 ◽  
Vol 20 (20) ◽  
pp. 131
Author(s):  
Elcí Terezinha Henz Franco ◽  
Cinara Echart Almeida
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Basal Medium ◽  
Optimal Combination ◽  
Coconut Water ◽  
Naphthalene Acetic Acid ◽  
Petiole Explants ◽  
Potato Extract ◽  
Whole Plants

Petiole explants of Begonia rex were cultured on basal medium (MURASHIGE & SKOOG, 1962). The medium was suplemented with naphthalene acetic acid (0.01; 0.1 and 0.5 mg/I) and kinetin ( 0.1; 0.2; 0.5 and 1.0 mg/I). In these experiments, were as also used coconut water (15% and 20%) or potato extract ( 15% and 20%). Buds were formed in several treatments, but the best combination was coconut water 0.01 mg/L NAA and 0.1 mg/I KIN. Whole plants (40% of the explants) were obtained when was added coconut water . The optimal combination for plant regeneration (100%) was 0.01 mg/I NAA plus 0.1 mg/I KIN.

COMPARAISON ENTRE DES LIGNEES PEDIGREES ET DES LIGNEES HAPLOIDES DOUBLEES CHEZ L’ORGE (HORDEUM VULGARE L.)

1980 ◽  
Vol 60 (1) ◽  
pp. 79-85 ◽  
Author(s):  
PIERRE TURCOTTE ◽  
C. A. ST-PIERRE ◽  
KEH MING HO
Keyword(s):  
Hordeum Vulgare ◽  
Doubled Haploid ◽  
Doubled Haploids ◽  
Block Design ◽  
Pure Line ◽  
Kernel Weight ◽  
Hordeum Vulgare L ◽  
Line Production ◽  
Doubled Haploid Lines ◽  
Time Required

Pedigree and doubled haploid lines from seven crosses of barley (Hordeum vulgare L.) were compared over 2 years. The lines were tested, in a randomized complete block design, in row plots at Ste-Foy in 1977 and in hill plots at Brawley, California in 1978. There are significant differences between the two methods of pure line production for grain yield, 1000-kernel weight, plant height, resistance to lodging and date of maturity. Furthermore, these significant differences between doubled haploid and pedigree lines seem to be tied to wide crosses. After showing that doubled haploids are superior for resistance to lodging, we conclude that the use of doubled haploid lines must be seriously considered in a barley breeding program on the basis of improved selection efficiency and the short time required to get homozygous lines.

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