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EN
Haploid plants are often used for genome mapping, to induce mutations and also for the production of homozygous plants. They are also needed in heterosis breeding. Androgenesis is one of the methods of their obtainment. The study focused on the development of induction haploid plants and double haploid lines of rye. In the experiment, seven varieties of rye were used: Amilo, Dankowskie Nowe, Dankowskie Z?ote, Kier, Motto, Walet, Warko. The seedlings were vernalised at natural conditions or in the refrigerator. Plants were grown in the greenhouse. Spikes with pollen at the uninucleate stage were harvested and stored at 4 C in the dark for 48 hours. The influence of the varieties and the medium used on the androgenesis was tested. Modified N6 and potato medium were used. Anthers were cultivated in the dark for 4 weeks at 28C. Very low incidence of callus-producing plants (0,0-0,3%) and of any regenerated plants was observed.
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vol. 38
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issue 4
415-424
EN
A large population of anther culture-derived barley regenerants and their progeny was tested for allele segregation at 1 isozyme and 8 morphological marker loci. The segregation of genetic markers was examined separately for haploid, diploid and polyploid regenerants. All the 9 analysed genes except al (albino lemma) on chromosome 3 segregated according to the expected 1 : 1 ratio in the microspore-derived barley population. There was no difference in allele distribution between haploid and diploid regenerants. Among the limited number of 34 analysed tetraploids a significant excess of the dominant allele at locus o (orange lemma) of chromosome 6 was also observed. The recombination frequency between linked genes (n - lk2 on chromosome 1 and r - s on chromosome 7) estimated in the DH population did not differ significantly from recombination rates calculated in F2 progeny or presented in barley chromosome maps. The phenomenon of gametic selection is discussed in relation to the genotype dependency of anther culture response and procedures used for DH production in barley.
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vol. 38
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issue 4
381-392
EN
With the numerous improvements in cereal tissue and wheat anther culture, it is necessary to determine which of the improvements should be combined for optimal response. This study was conducted using one highly responsive cultivar of wheat (Triticum aestivum L. cv. Pavon 76) to test the effectiveness of pre-culture cold treatment (0 or 3-7 days at 5oC) of anthers, five initiation basal media, and various changes in Murashige-Skoog regeneration media. A cold pre-culture treatment was inhibitory for all initiation media for embryoid initiation. Of the initiation media, P1, 85D12, and N6 were similar for embryoid initiation (0.80 to 0.90 embryoids/anther) without a cold pre-culture treatment. Plant regeneration was improved by the addition of amino acids or glucose, increased sucrose concentration, filter sterilizing the medium, and altering plant growth regulator concentrations. P1 medium which is normally used for embryoid initiation was also beneficial for improving plant regeneration. Ethylene inhibitors were generally not beneficial.
EN
Anther and somatic tissue culture in combination with mutagenesis were carried out to evaluate the efficiency of different mutagenic treatments of various in vitro culture materials, and to obtain some promising variants for rice improvement. Results indicated that in japonica rice radiation treatment of dry seeds and young panicles influenced the percentage of green plantlets regeneration from anther culture. Both treatments increased significantly the percentage of regenerated green plantlets in comparison with the control. Irradiation with 30 Gy of rice callus increased also the percentage of regenerated green plantlets. For indica rice, the combination of the suitable dose of gamma rays irradiation on seeds and an improved medium, increased the percentage of callus induction. This approach made it possible to use anther culture in indica rice breeding. Somatic tissue cultures combined with radiation-induced mutagenesis led to the development of a number of promising mutants including some new cytoplasm-nucleus interacting male-sterile lines with almost 100% stigma exsertion. Their development would be of practical significance for increasing the genetic diversity for production of hybrid rice.
EN
The aim of this study was to evaluate phenotypic variation of R0 androgenic plants obtained from four seed sources and donor plants by anther culture. Several morphological traits (leaf size, petiole length, leaf division, cortex colour) and the range of diversity were evaluated. There was large variation in all traits among the donor varieties. Especially leaf division and cortex colour differed significantly among the androgenic plants that came from different seed sources. The plants regenerated from four donor plants of variety 62 were significantly different in most traits except for leaf width and cortex colour. Evaluation of R1 plants will demonstrate whether the R0 variation observed is due to genetic variation or physiological differences from tissue culture.
EN
Induction of androgenesis in in vitro microspore culture of barley is the most efficient method of haploidisation, but it is difficult to use in the conditions of a plant breeding station. For this reason, the method has been improved side by side with the technique of anther culture. In anther culture of 3 barley cultivars, pretreatment with 0.7 mol dm-3 mannitol and the addition of 0.2 mg dm-3 2,4-D to the induction medium have been shown to exert a favourable effect. In cv. Mobek, the interaction of the two tested stress agents, mannitol and auxin, has been proved.
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vol. 38
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issue 4
425-435
EN
F1, F2 populations as well as F3 and F4 wheat lines coming from more than 300 cross combinations have been used for androgenic wheat studies and more than 5000 haploid plants were produced. Promising doubled haploid lines were tested together with lines produced in the conventional way in yield trials. The two best doubled haploid lines overyielded the standard. On the basis of results related to the general combining ability it is advisible to cross parents with a good combining ability for characters such as high percentage of responding anthers and embryo/callus induction with parents indicating a good combining ability for green plant production. In this way ideal lines can be obtained for doubled haploid production.
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vol. 38
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issue 3
253-258
EN
Anthers of twenty triticale genotypes were cultured on three different media: 1 - PII (CHUANG et al. 1978) with increased 2,4 D to 2 mg L 1 and agarose 6 g L 1; 2 - Macro , micronutrients and vitamines like in MN6 (CHU, HILL 1988) + 2 mg L 1 2.4 D + 0.5 mg L 1 KIN + 5 mg L 1 FeEDTA + 90 g L 1 sucrose; 3 - Macro , micronutrients and vitamines like in MN6 (CHU, HILL, 1988) + 2 mg L 1 2.4 D + 0.5 mg L 1 KIN + 5 mg L 1 FeEDTA + 120 g L 1 sucrose. Embryoid induction and plant regeneration were influenced by donor plant genotype and induction medium. Medium 1 was the best for embryoid induction, while for green plant regeneration the best were media 1 and 2. Out of 300 anthers from each genotype plated on each of the three media, 64 1250, 12 486 and 6 212 somatic embryos and 8 86, 3 136 and 1 26 green plants were recorded, on media 1, 2 and 3, respectively.
EN
Several aspects of in vitro culture have potential for cereal improvement. This paper focuses on evaluation of somaclonal variation (SV) from immature embryo callus culture, and doubled haploid (DH) production via anther culture in barley. Genetically stable SV was observed for several seedling morphological traits such as albino, yellow, light green and lethal. SV occurred at approximately half the frequency of azide-induced mutagenesis. The potential for widespread application of anther culture-mediated DH production in barley breeding and genetic studies was increased through culture procedure improvements and understanding the inheritance of anther culture response. Methodology improvements included substitution of inexpensive gelrite for expensive ficoll or agarose, ability to grow anther donor plants under field as well as growth chamber conditions and flexibility in cold pretreatment/storage of anther donor spikes for 4-6 weeks prior to anther plating. From diallel analysis, inheritance of anther culture response was complex with additive and dominance effects for embryoid formation, total plant regeneration and green plant regeneration and reciprocal effects (maternal) for green plant regeneration. High x low responder crosses generated F1's that were intermediate in response and low x low crosses sometimes produced F1 heterosis for green plant regeneration. Therefore, some recalcitrant types appear to be usable in anther culture DH production systems within a breeding program.
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vol. 38
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issue 4
407-414
EN
The use of 2,4-D as a post pollination treatment to fertilize florets instead of GA3 provided a two-fold improvement in seed set, culturable embryos and hence green plant production from wheat and maize pollination. The efficiency of the wheat by maize pollination method for haploid production was equal to the anther culture method in F1 hybrid combinations that were responsive to anther culture. However, in recalcitrant combinations obtained from non-responsive genotypes the maize pollination method was far superior.
EN
Miscanthus sinensis is a promising species for biomass production. Influences of genetic and nongenetic factors on androgenesis induction efficiency were investigated. This is the first report on successful induction of pollen-derived callus in M. sinensis. The callus yield was strongly affected by genotype. A beneficial influence of cold pretreatment of spikes on androgenesis induction was observed. The highest yield of calli was obtained in cultures on a modified C17 medium. The results suggest that the high callus yield might be caused by the late culture initiation. The beginning of anther culture at the end of the flowering season caused a 17-fold increase in callus yield, in comparison to culture initiated at the height of the flowering season (August). It is likely, however, that the efficiency of androgenesis induction in the case of M. sinensis anther culture beginning in October could be related to a positive influence of growing donor plants in conditions of cooler and shorter day, i.e. 11-h day with temperature around 11oC and 13-h night with temperature around 5oC. Results of this study can significantly support the development of effective methods of M. sinensis haploidization, which could be used in crop improvement by breeding.
EN
The effect of cold pretreatment of spikes on somatic embryo induction and ather culture response of 25 F1 winter hybrids was investigated.The efficiency of androgenic embryos was the highest when spikes were incubated at 4 C degree for 6-9 days.A total of 2242 (73%) green and 885 (27%) albino plants were obtained from 9900 cultured anthers.Anther culture response in wheat wsa found to be markedly affected by the genotype of donor plants.This percentage of green plants varied from 0 to 115.7%.The great majority of anther-derived regenerants were haploids (82.35%), while the remaining plants were spontaneous diploids (13,73%) and aneuploids (3.92%).
EN
Tolerance to a new herbicide, pyributycarb, was evaluated both at the plant and cellular levels. Several highly or moderately tolerant strains chosen at the plant level, showed a parallel relation of to tolerance at the cellular level. However, on the whole, correlation between total tolerance indices and survival rates of calli was not significant in 18 out of the 80 studied strains. As a result of somaclonal selection for two herbicides, lines NB-200 and NM-100 were regenerated from the tolerant calli screened with benthiocarb at 200 ppm, and molinate at 100 ppm, respectively. In the R2 generation, both the lines displaned a stable tolerance both at the plant and cellular levels. Thus the highly tolerant mutant lines were developed from a moderately tolerant line, N-61, via in vitro selection. To achieve a short-cut method in the interspecific genetic exchange, a series of techniques related to cell fusion were established in rice and related species. Two kinds of somatic hybrids between the cultivar Kitaake and tetraploid Oryza species, O. punctata and O. officinalis, were successfully produced. Among the somatic hybrid plants, a wide range of chromosomal variation was observed. Aneuploid plants with a chromosome number around 2n = 72 (hexaploid), which are expected from a symmetric fusion between diploid and tetraploid strains, were obtained showing mixoploidy within a plant. Most of the somatic hybrids were characterized by intermediate features of plant-type showing high sterility, shattering of spikelets and reduced plant height. As an exception, a diploid plant, which was identified by RFLP analysis using the rDNA gene probe, closely resembled Kitaake and produced viable seeds. A tetraploid hybrid plant was also promising for the introduction of economically important characters through the reduction of chromosome numbers by doubled haploids. Gametoclonal variation and gamma radiation was applied to Kitaake. The mutation frequency was prominently increased by gamma ray treatment, especially at high doses of 200 Gy or 300 Gy. In the M3R2 or M4R3 generations, most of the variants showed unfavourable characters. Most of the mutant characters were governed by single or double recessive genes. Several mutants such as short culm and early flowering time might be used for rice breeding.
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