A cytological study of pretreatments used to improve isolated microspore cultures of wheat (Triticum aestivum L.) cv. Chris

Genome ◽  
1999 ◽  
Vol 42 (3) ◽  
pp. 432-441 ◽  
Author(s):  
Tianci Hu ◽  
Ken J Kasha
Keyword(s):  
Nuclear Division ◽  
Callus Formation ◽  
Microspore Culture ◽  
Triticum Aestivum L ◽  
Induction Medium ◽  
Isolated Microspore Culture ◽  
Large Numbers ◽  
Isolated Microspores ◽  
Uniform Population

Isolated microspores of wheat can be induced in vitro to switch their development from the gametophytic pollen pathway to a sporophytic pathway, resulting in embryoid or callus formation. The influence of cold or mannitol pretreatment on karyokinesis and cytokinesis in isolated microspore culture responses were investigated. Anthers were pretreated in mannitol for 7 d at 28°C; spikes at 4°C for 28 d. Microspores often completed the 1st mitotic nuclear division during pretreatment while cytokinesis was delayed. During mannitol pretreatments, the 1st mitotic nuclear division was mostly symmetrical while only asymmetric 1st nuclear divisions were seen during or after cold pretreatment. Following the symmetrical division, the two similar nuclei often appeared to fuse to form a diploid nucleus. Subsequently, these nuclei underwent rapid nuclear divisions to form multinucleate, and later, multicellular structures in induction medium. Cold pretreatments also induced muticellular structures but frequencies were lower than after mannitol. A novel pretreatment of spikes, combining 0.4 M mannitol solution at 4°C for 4 d, delayed the 1st nuclear division, keeping all microspores in a haploid uni-nucleate stage and resulted in higher induction frequencies. The proportion of embryos larger than 2 mm that developed into green plants was as high as 70% when transferred to regeneration media. Ninety-five percent of the plantlets transferred from culture to soil survived. The improved pretreatment enhanced the potential of isolated microspore culture in wheat for plant breeding by producing large numbers of plants and for gene transformation by maintaining a uniform population of haploid uni-nucleate stage microspores as targets.Key words: wheat, pretreatment, karyokinesis, embryogenesis, microspore, cold, mannitol.

Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee
Keyword(s):  
Winter Wheat ◽  
Chinese Spring ◽  
Shoot Growth ◽  
Callus Formation ◽  
Triticum Aestivum L ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

scholarly journals Improvement of methods of creating hybrids of cabbage

2019 ◽  
pp. 3-7 ◽  
Author(s):  
Anna I. Mineykina ◽  
Lyudmila L. Bondareva ◽  
Darya V. Shumilina ◽  
Elena A. Domblides ◽  
Alexey V. Soldatenko
Keyword(s):  
First Generation ◽  
Microspore Culture ◽  
Diallel Cross ◽  
High Yield ◽  
Isolated Microspores ◽  
Economically Valuable Traits ◽  
Complete Diallel Cross ◽  
Speed Up ◽  
Pure Lines

Relevance One of the basic directions of the cabbage crop breeding is the creation of F1 hybrids with a complex of economically valuable traits. This process is difficult and time-consuming as to get pure lines must be within 6-12 years hold inbreeding. Herewith not every line gives the desired heterotic effect that also requires additional verification. Methods Biotechnological method culture of isolated microspores in vitro, which allows in the first generation to receive a line with 100% homozygosity, was used to speed up the breeding process. Combination ability were performed in complete diallel cross on the basic morphological signs. Results Culture medium for cultivation of isolated microspores in vitro was optimized for each genotype of cabbage for the best embryoids regeneration. Maximum amount of embryoids was received on medium with pH 6.2 using ampicillin 100 mg/l and zeatin 1 mg/l: 466.7 ± 153.2 PCs/100 buds. A new source material for breeding – doubled haploid lines of cabbage was received. Lines – the best parents for F1 hybrids with high yield, compact rosette of leaves, with optimum inside and short outside cabbage stump was created. Studies have shown that optimization of breeding process in case of creation of pure lines of cabbage in 3 years with microspore culture requires to reduce the breeding process in 2 times.

IN VITRO ANTHER AND ISOLATED MICROSPORE CULTURE AS TOOLS IN SWEET AND SPICE PEPPER BREEDING

2009 ◽  
pp. 61-64 ◽  
Author(s):  
A. Gémes Juhász ◽  
Z. Kristóf ◽  
P. Vági ◽  
C. Lantos ◽  
J. Pauk
Keyword(s):  
Microspore Culture ◽  
Isolated Microspore Culture ◽  
Pepper Breeding

scholarly journals 882 PB 276 IN VITRO PROPAGATION AND CHIMERAL TRAITS OF Cryptanthus `Marian Oppenheimer' (WIDE LEAF CLONE)

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 560c-560
Author(s):  
Yong Cheong Koh ◽  
Fred T. Davies
Keyword(s):  
In Vitro Propagation ◽  
Callus Formation ◽  
Adventitious Shoot ◽  
Shoot Formation ◽  
Induction Medium ◽  
Ms Medium ◽  
Periclinal Chimera ◽  
Adventitious Shoot Formation ◽  
L1 And L2

The leaves of vegetative stolons of greenhouse grown Cryptanthus `Marian Oppenheimer' (wide leaf clone) were cultured in modified MS media to induce adventitious shoot formation via callus formation. The best callus induction medium was basal MS medium with 10 μM NAA, IBA and BA. Pure green (843), maroon (3), striped (2) and albino plantlets were obtained. Most of the albino plantlets were stunted, tightly clumped together and impossible to score. The medium which produced the highest average number of non-albino plantlets was basal MS medium with 0.3 μM NAA, IBA and BA All non-albino plantlets were rooted in MS medium with 5.4 μM NAA and transplanted ex vitro with a survival rate of 96.7%. The maroon plantlets became green two weeks after transplanting. Histological studies revealed that C. `Marian Oppenheimer' (wide leaf clone) has two tunicas (L1 and L2) and a corpus (L3). Callus on the leaf explant arose mainly from the L2 and L3. Apparently C. `Marian Oppenheimer' (wide leaf clone) is a GWG periclinal chimera.

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 Studies on In Vitro Response to Callus Induction and Regeneration of Five High Yielding Indica rice Varieties

2019 ◽  
Vol 7 ◽  
pp. 97-104
Author(s):  
Md. Niuz Morshed Khan ◽  
Md. Monirul Islam ◽  
Dr. Md. Shahidul Islam
Keyword(s):  
Callus Induction ◽  
Callus Formation ◽  
Indica Rice ◽  
Induction Medium ◽  
Embryogenic Calli ◽  
Rice Varieties ◽  
New Variety ◽  
Increasing Demand ◽  
Embryogenic Callus Formation

Due to growing population, there is an increasing demand of rice production but the productivity of rice is lessened day by day. To overcome this problem various biotechnological tools can be used for developing various rice varieties. However, the lack of a simple and efficient protocol for callus induction, embryogenic callus formation and quick plant regeneration in this cereal crop. In this study embryogenic calli from mature seeds of five indica rice varieties viz. Binadhan-5, Binadhan-6, BRRI dhan-48, BRRI dhan-58 and IR-64 were observed that is done in four different types of media composition. The highest callus induction were observed in media containing Sucrose as a carbon source. Among those varieties Binadhan-6 and BRRI dhan-48 showed highest rate of callus induction respectively. This study will be useful for selecting suitable callus induction medium for callus induction in future that will be useful for not only national but also international plant breeders for producing new variety and so on.

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 CULTURE USE PROBLEMS IN SELECTION OF ISOLATED MICROSPORES IN GRAIN

2020 ◽  
Vol 55 (1) ◽  
pp. 31-39
Author(s):  
G. A. Iskakova ◽  
D. O. Baisaparova ◽  
D. R. Raimbek ◽  
K. Zh. Zhambakin
Keyword(s):  
Genetic Basis ◽  
Selection Process ◽  
Microspore Culture ◽  
Wheat Breeding ◽  
Identification System ◽  
Cultivar Development ◽  
Isolated Microspores ◽  
New Gene ◽  
Selection Of

Production of haploid plants by culture of isolated microspores is a quick way of obtaining homozygous crop lines. Recessive features of mutant homozygous plants are also possible to determine by this biotechnology. Contrary from anthers culture, in which the presence of anther walls can lead to the development of diploid somatic calli and plants, the microspore culture produces only haploid or dihaploid lines. Isolated microspores culture in addition represents and has a unique identification system for studying the mechanisms of embryogenesis in in vitro culture. The usage of haploid technology extends the genetic basis of wheat breeding, since it allows increasing the frequency of new gene combinations. This technology significantly increases the efficiency of breeding new highly productive varieties of crops. On this basis, it becomes possible to quickly assess the prospects of dihaploids, which significantly improves the efficiency of the selection process. DH plants are completely fertile and, if necessary, may be used as parents or processed as a cultivar. DHs have been widely used for cultivar development, genetic mapping, mutagenesis, and the study of gene functions.

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