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1.
Flower buds and extracted anthers of two apple genotypes, ‘Golden Delicious’— a previously known non-androgenic genotype — and ‘Topred’— a known androgenic genotype —, were irradiated by gamma-rays at doses ranging from 5 to 20 Gy, after 0 to 5 weeks of cold pre-treatment (3 °C). When the extracted anthers were irradiated, both microspore embryogenesis and callogenesis were limited for the two genotypes. For ‘Golden Delicious’, androgenic embryogenesis was induced from the flower buds irradiated at 10 to 20 Gy and after 3 to 5 weeks of cold pre-treatment. For ‘Topred’, androgenic embryogenesis was slightly improved after flower bud irradiation at 5 to 10 Gy following cold pre-treatment from 0 to 3 weeks; it was limited after 5 weeks of cold pre-treatment. 相似文献
2.
Anna Pretova Norbert C.A. de Ruijter André A.M. van Lammeren Jan H.N. Schel 《Euphytica》1992,65(1):61-69
Summary The capacity of the maize genotype 4c1 to regenerate microcalli and embryos from cultured microspores has been examined by comparing various cold pretreatments and culture media, using microspores and pollen at different stages of development. Viability of cultured cells was tested with FDA and their development was traced with light and fluorescence microscopy using DAPI as a nuclear dye.It was found that a pre-incubation of dissected flowers floating in a liquid nutrient medium at 8°C during 10–14 days was most successful for the induction of cell division. Among the developmental stages tested only the microspores appeared to regenerate. Subculture at 25°C in the same liquid medium, supplemented with 0.1 mg/l TIBA, gave highest rates of microspore division, i.e. up to 70% at 4 to 6 days of culture.All pathways described earlier for maize androgenic embryogenesis were observed within the 4c1 genotype. Symmetric divisions occurred in cultured microspores but most frequently asymmetric divisions lead to the formation of microcalli within 12 days of culture. In at least 60% of all dividing microspores cells were derived from the generative nucleus. Microcalli further developed either into loose or compact calli. Compact calli formed embryo-like structures.Abbreviations DAPI
4,6-diamidino-2-phenylindole
- Dicamba
3,6-dichloro-2-methoxy benzoic acid
- 2,4D
2,4 dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- PAA
phenylacetic acid
- TIBA
2,3,5-triiodobenzoic acid
- YP medium
Yu-Pei basal salt medium 相似文献
3.
Summary Callus culture of immature wheat-rye hybrid embryos was compared with embryo culture in two experiments. Embryos were rescued from field grown mother plants at two day intervals 13–21 days after pollination and plated for 1) callus culture on Murashige and Skoog medium (MS) supplemented with 2 mg/12,4-dichlorophenoxy acetic acid followed by plant regeneration on hormone free MS medium with half strength mineral salts, 2) embryo culture on Taira and Larter medium (TL). Observations were made on embryo size and condition at time of rescue (Experiment 1) and embryo development directly into plants (embryo culture) or through embryogenesis (callus culture). Fewer 19 and 21 day old embryos developed into plants from callus culture than from embryo culture in Experiment 1. Callus culture was more efficient than embryo culture in promoting plant recovery from 17 day old embryos in Experiment 2. The number of plants per embryo was significantly higher from callus culture than from embryo culture. In both experiments callus culture promoted embryogenesis in more embryos than developed in embryo culture. Embryo rescue 15–17 days after pollination was optimal in both experiments. 相似文献
4.
In linseed, the time needed to obtain pure lines can be reduced by using in vitro anther culture. The technique has been improved but there are no reports on the inheritance of the characters (callogenesis and regeneration) involved in the response. To shed some light on this aspect, anthers of four linseed varieties and their F1 were cultivated in vitro. Callogenesis response was analysed according to a full diallel model. Among 7811 anthers, the percentage of callogenesis ranged from 27.40 to 82.04%, with the parental lines being around the average. The differences between specific hybrid combinations were significant and general and specific combining ability were highly significant. As well as dominant effects, additive and non‐additive effects are probable components of the heritability of this character, although additive effects would be more important than non‐additive. Differences between reciprocal crosses would indicate that cytoplasmic factors could also be involved in the determination of the character. 相似文献
5.
Summary Microspore culture was shown to be applicable to a broad range of accessions belonging to six horticulturally important crop types of Brassica oleracea: broccoli, white cabbage, cauliflower, savoy cabbage, Brussels sprouts and curly kale. Of 64 accessions tested 86% were responsive. Large genotypic differences were found in number of embryos produced per flower bud, and in frequency and mode of regeneration of plants from embryos.
B. oleracea was characterized by a strong asynchrony of microspore development within single buds. Microspore populations optimal for culture contained a large proportion (10–40%) of binucleate pollen. An initial high temperature treatment was essential for microspore embryogenesis. Growth conditions of the donor plants during inflorescence formation were less critical. 相似文献
6.
Summary Embryo age and composition of nutrient medium affected plant growth and response to vernalization in winter wheat (Triticum aestivum L.). Root and shoot development was more in older than in younger excised embryos, and more in a medium without kinetin than in one with kinetin. Kinetin (2 mg/l) in the medium did not accelerate vernalization, probably because it tended to inhibit seedling and plant growth.Embryo age and media did not completely replace vernalization. Twenty- and 16-day-old embryos responded by flowering after 4 weeks of vernalization. Among plants raised on a standard medium from 20-day-old embryos and vernalized for 4 weeks, 84.2% flowered by or before 50 days after transplanting. Time from embryo culture to heading for 20-day-old embryos with-4-week vernalization averaged 84.6 days. Immature embryos (16–20 days old) needed only 4 weeks of vernalization compared to 6 weeks for mature embryos. Excised embryos could be vernalized as efficiently as seedlings raised by embryo culture. Embryo culture at 16–20 days after anthesis coupled with 4-week cold treatment shortens generation time of winter wheat by about 40 days.Contribution No. 82-131-j, Department of Plant Pathology, Kansas Agricultural Experiment Station, Manhattan, KS 66506, USA. 相似文献
7.
Serge Gudin 《Euphytica》1993,72(3):205-212
Summary Two crosses between Rosa hybrida L. cultivars, that fail to produce progenies by conventional means, were carried out. In one of them, total hip abscision occured seven weeks after pollination; in the other one, only 2% of the fertilized hips remained on the plants after eleven weeks. Four- and five-week-old fertilized ovules were isolated in the first cross and six-week-old embryos in the second. The ovules were cultured on six media which differed in their mineral salt and sucrose concentrations while the embryos were cultured on only one of these media and eventually cold treated for one month at 4° C before being placed in a culture room set at 23° C. The embryos that had been isolated in ovulo when they were still not visible under a binocular lens developed atypically. The embryos isolated in ovulo when they were heart-shaped and on average 0.27 mm long performed in ovulo germination on some media and/or enlargement on all of them after two weeks; after another two week culture, once isolated from the ovule, all of them germinated. The cultured isolated embryos, that were exposed to cold, enlarged and germinated more rapidly when placed at 23° C than those not exposed to cold. Furthermore, the plantlets resulting from untreated embryo germination were characterized by large cotyledons which were only partially green. These results are discussed in regard to embryogenesis, precocious germination and dormancy. 相似文献
8.
A protocol for high frequency callus induction and plant regeneration from sunflower (Helianthus annuus L.) anthers is described.
Different variables using Murashige & Skoog (MS) basal medium supplemented with 2.0 mg/l α-naphthaleneacetic acid (NAA) and
1.0 mg/l N6-benzyladenine (BA) were tested for their ability to enhance the frequency of anther callusing and subsequent embryogenesis.
Of these, agar concentration, sucrose concentration, carbohydrate source had significant effect on callusing, while differences
due to incubation under dark vs light conditions, cold pretreatment of capitula for 1 to 6 days prior to anther inoculation
and genotype on callusing were non-significant. However, all these factors exerted highly significant influence on embryogenesis
when calli from the various media were transferred to medium supplemented with 0.1 mg/l NAA and 0.5 mg/l BA. With the procedure
developed, callusing as high as 100% and embryo formation at a frequency of 44% was achieved. Although complete embryos were
formed the frequency of their conversion to whole plantlets was low (14.3%). Hence, the embryogenic pathway was bypassed to
obtain multiple shoots by transferring embryogenic calli with developing embryos to MS medium supplemented with 0.5 mg/l BA.
Elongated shoots rooted on half-strength MS medium supplemented with 0.5 mg/l NAA. Cytological analysis of embryogenic callus
and somatic embryos revealed haploids at a frequency of 30% while that of rooted plants showed haploid regenerants at a frequency
of 8.3%. Nevertheless, the frequency of putative haploid plants could be enhanced through mass multiplication using nodal
explants of the regenerants.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
9.
Effect of activated charcoal on Brassica oleracea microspore culture embryogenesis 总被引：11，自引：0，他引：11
João Carlos da Silva Dias 《Euphytica》1999,108(1):65-69
The effect of the addition of a 0.1 ml drop of activated charcoal (AC) on microspore culture embryogenesis was studied in
nine morphotypes of Brassica oleracea. Embryo yields were significantly increased in all of the morphotypes by the addition
of the 0.1 ml drop of AC to the microspore culture media. The magnitude of the response to the addition of AC varied with
the different plants and morphotypes. The addition of AC never produced a detrimental effect. A qualitative improvement of
the subsequent development of embryos to plants was also observed with the addition of AC. Data suggest that the addition
of AC to microspore culture media promoted embryo production in different B. oleracea morphotypes.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
10.
Effects of donor plant temperature,photoperiod, and age on anther culture response of Capsicum annuum L. 总被引：7，自引：0，他引：7
Summary Effects of temperature and photoperiod for donor plant growth on embryo formation in Capsicum annuum anther culture were investigated. Donor plants were grown in glasshouses at minimum temperatures between 16 and 30° C and at photoperiods between 11 and 19 h. Anthers were collected from individual plants over five to nine week periods to test the significance of donor plant age. Embryos were obtained at all temperature regimes with a calculated optimum temperature of 26.4° C. Embryo formation was unaffected by the photoperiods tested. Embryo formation varied among successive samplings. However, a significant decline in anther culture response with increasing donor plant age was observed in all three experiments.Abbreviations AT
average air temperature the week before anther incubation
- EMB 100
1n (embryos per 100 anthers +1) 相似文献
11.
The effect of colchicine on induction of embryogenesis andchromosome doubling during microspore culture was evaluated in twoF1 hybrids of spring oilseed rape (Brassica napus L.). Immediatecolchicine treatment of isolated microspores with the concentrations 50 and500 mg/L for 15 h stimulated embryogenesis and produced largeamounts of healthy-looking embryos. These normal embryos germinatedwell at 24 °C after being transferred to solid regeneration mediumand an initial period of low temperature (2 °C) for 10 days, andcould directly and rapidly regenerate vigorous plants. A high doublingefficiency of 83–91% was obtained from 500 mg/L colchicinetreatment for 15 h with low frequency of polyploid and chimeric plants.The present experiment showed that a treatment duration of 30 h revealedless positive effects on embryogenesis and doubling efficiency, especially athigher colchicine concentration (1000 mg/L). Poor embryogenesis andembryo germination were observed from ordinary microspore culturewithout change of induction medium and colchicine treatment, and severalsubcultures were required for induction of secondary embryogenesis andplant regeneration. 相似文献
12.
B. S. Ahloowalia 《Euphytica》1994,75(3):163-172
Summary Production of mini-tubers as a source for seed potato was investigated by growing in soil micropropagated plants and micro-tubers produced from micropropagated plants. Cultures of several cultivars were initiated from indexed tubers and multiplied on modified MS medium. Cultures were micropropagated by using a modular system which allowed batch handling. Micropropagated plants produced mini-tubers in glasshouse after 70–115 days of growth in soil. A large proportion of the mini-tubers produced were between 9 and 15 mm diameter. Several factors, e.g., explant number, duration of in vitro culture and genotype influenced mini-tubers production. Micropropagated plants after culture of 86 days or longer produced micro-tubers ca. 2 to 10 mm diameter. Plants, which formed micro-tubers in vitro, produced less number of mini-tubers in soil. Micro-tubers produced 1 to 3 mini-tubers when grown in soil in chain-type paper pots, but produced conventional sized tubers when grown in soil under plastic polytunnel. Mini-tuber number varied widely between potato cultivars; cvs. Bintje and British Queen produced more mini-tubers than the other cultivars. Mini-tubers developed green hard skins when kept in light for 3 weeks, and could be stored in dark at 4° C upto 6 months. In a field trial, small mini-tubers ca. 5–10 mm diameter produced more but smaller tubers than mini-tubers ca. 15–20 mm diameter. The micropropagated plants and the plants grown from mini-tubers were genetically stable, and did not show any morphological aberrations except for one variegated plant among the several thousand produced. It is concluded that the production of mini-tubers by soil planting of micropropagated plants is a rapid and efficient method for producing seed potato tubers.The use of any trade mark and other commercial products mentioned in this paper does not constitute any recommendation or advertisement for such products on behalf of the Agriculture and Food Development Authority, Dublin, Ireland 相似文献
13.
Summary The effect of direct vernalization of immature embryos on flowering was studied in six winter wheat genotypes. Fourteen-, 17-, and 20-day-old embryos were excised and vernalized for 0–6 weeks on synthetic medium during a conditioning period. Percent germination of embryos was high (overall 96.1%), and free from genotypic effects. Genotypes differed for flowering in response to cold treatment of excised embryos. Embryo vernalization was as effective as or more than conventional vernalization (control, seedling vernalization for 6 weeks). Seventeen-day-old embryos were the most responsive to vernalization. With a 5-week vernalization of 17-day-old embryos, the percentage of plants anthesed was higher than those from 14-and 20-day-old embryos. For 17-day-old embryos vernalized for 5 weeks, the mean number of days from culture to anthesis was less than that of 6 week vernalization, less than that of 14- and 20-day-old embryos, and less than controls.Purdue Univ., Agronomy Dept., W. Lafayette, IN 47907, USA. 相似文献
14.
Summary Pollen grain embryogenesis in anther cultures of Brassica juncea cv. PR-45 was considerably enhanced by treating the donor plants with 4 mll-1 (v/v) of ethrel or delayed sowing of the donor plants, the latter treatment being superior. The anthers derived from plants sown about two months after the normal sowing period showed 18% androgenesis as compared to 3.5% in the control.Pollen grain embryos normally showed very poor germination (10%) on B5 or B5 containing GA3. However, ABA or cold treatment promoted normal germination of these embryos. Exposure of the embryos to 4°C for 6 days, which proved to be the best treatment, induced 66% germination of the embryos. 相似文献
15.
Summary Interspecific crosses between Cucumis metuliferus Naud. and C. anguria L. were obtained through embryo culture. Embryos in the rabbit-ear to advanced fluke-shaped stages were rescued 34–99 days after pollination. Plants were obtained through direct embryo culture, and through somatic embryogenesis from immature embryos. For direct embryo culture, fluke-shaped embryos were stored in sterile water in darkness for three days at 25C prior to transfer on Murashige and Skoog (MS) culture medium plus 1.0 M 6-benzylamino-purine. Multiple plants were obtained from single embryos through somatic embryogenesis of rabbit-ear stages on MS plus 10 M indole-3-acetic acid and 5 M 6-benzylamino-purine. Evidence of hybridization included leaf shape intermediate between the two parents, penduncle shape prior to fertilization which resembled the male parent, low pollen viability and isoelectric focussing of protein bands for acid phosphatase of leaf extracts. 相似文献
16.
Anther culture of an interspecific rice hybrid from a cross of Oryza sativa× O. rufipogon was attempted. Of the 117 regenerated pollen clones, 56 could survive to maturity. A majority of these were either haploids or doubled haploids and very few turned out to be chromosomal variants. Comparative study of doubled haploids and the seed derived F2 plants indicate the distinct advantages of anther culture techniques. (1) Androgenic plants, though few in number, showed greater ariation for all the traits with the exception of ear bearing tillers. (2) Predominance of recombinants with wild traits was observed in F2 segregation. (3) It was possible to recover indica type recombinants among the anther-derived plants with one or two traits introgressed from O. rufipogon. These results suggest the feasibility and utility of anther culture in distant hybridization for incorporation of alien variation into cultivated rice. 相似文献
17.
几丁质酶及β-1,3-葡聚糖基因转化橡胶树的研究 总被引：1，自引：1，他引：0
旨在通过基因工程的方法提高橡胶树白粉病抗性。以高产、综合性状优良的大规模推广级品种‘热研7-33-97’15090块花药愈伤组织为受体,通过农杆菌介导将菜豆几丁质酶和烟草β-1,3-葡聚糖酶基因转入橡胶树,获得1386个抗性胚状体,其中277个通过次生体胚发生成功增殖。PCR结果表明有49个胚状体为阳性,转化效率为0.33%。本研究通过农杆菌介导将菜豆几丁质酶和烟草β-1,3-葡聚糖酶基因转入橡胶树,PCR阳性率为0.33%,为通过基因工程手段提高橡胶树白粉病抗性奠定了良好的基础。 相似文献
18.
Somatic in vitro culture response of Lolium perenne L.: genetic effects and correlations with anther culture 总被引：2，自引：0，他引：2
Summary Genotypic effects on callus induction and plant regeneration in callus, suspension and protoplast culture, and their correlations with both phenotypic and GCA-values for anther culture response, were studied using 21 genotypes of perennial ryegrass. Differences between genotypes accounted for approximately 40% of the total variation for callus induction and initial callus growth, and 59 and 83% of the variation in callus culture for regeneration percentage and percentage of green plants. Effects of genotypes were less pronounced in suspension culture, where suspensions from the same genotype often behaved differently. Some suspension cultures retained their capacity for green plant regeneration for almost two years, repeatedly producing 80–100% green regenerants during this period. Genotypes with high regeneration percentage and a large proportion of green plants from callus culture were also superior in suspension culture for both regeneration performance and longevity. Regeneration percentage and percentage of green plants were uncorrelated, and probably under different genetic control. While capacity for green plant formation from the different genotypes showed no correlation between anther culture and somatic in vitro culture, a positive correlation was observed between the regeneration percentages in somatic in vitro culture and anther culture (r=0.44*–0.85***), suggesting some common genetic control of the two systems. 相似文献
19.
The genotypic responsiveness to androgenesis and the effect of two exposure times of cultured anthers to auxins (12 days and the entire period of culture) were studied in three lines of cauliflower (one spring - and two winter-types). The anthers of all genotypes responded to the protocol by producing embryos (0.2 -25.3%), 44.2% of which regenerated plantlets through organogenesis on a regulator-free B5 medium containing 40 g/l of sucrose and 800mg/l of L-glutamine. Embryo yield improvement, recently achieved on solid medium with the addition of 125mg/l silver nitrate, was not observed in liquid cultures. Mean frequencies of androgenic embryos, as affected by the duration of auxin supply to the culture, were always significantly higher (6.6- and 8.1-fold.) in the‘12 days of exposure’treatment than in the‘entire period of culture’treatment. The resolution of the parameter‘embryo percentage’into its two components (% of responsive anthers and number of embryos per responsive anther) gave evidence of a highly significant increase in the percentage of responsive anthers, whereas the number of embryos per responsive anther did not change statistically. The percentage of embryos regenerating plants was not influenced by the duration of auxin supply during anther culture. Chromosome count analysis revealed a haploid/diploid/polyploid plant ratio of 6:79:15. Of the 78 androgenic plants (RO generation) grown in the field, only 14 were fertile and produced seeds. RAPD analysis showed that none of the seven polymorphic loci out of 23 analysed expressed polymorphism within the RI progenies of 11 anther culture-derived lines. 相似文献
20.
Summary Shoot base segments have been explanted from seedlings of rice (Oryza sativa L. subsp. Japonica, cv. Arborio) and grown on agar-solidified MS medium supplemented with different concentrations of four cytokinins: kinetin, BAP, 2iP and zeatin. After one month, segments were explanted from proliferated shoots and subcultured on their respective media. BAP was by far the most effective in inducing shoot proliferation. Highest rates were achieved at the higher concentration used: 5 mg 1–1. Shoot base segments were subcultured fifteen times consecutively on seven different concentrations of BAP. Shoots grown in the presence of 5 mg 1–1 of BAP proliferated an average of 12 normal shoots for each base segment throughout the fifteen subcultures. The shoots rooted easily on hormone-free medium. The technique does not require any particular skill, it is very effective and, therefore, can be suggested as suitable for clonal propagation of rice. 相似文献