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  • 1
    ISSN: 1573-5060
    Keywords: Trifolium repens ; white clover ; callus ; genotypic variation ; protoplast ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary Callus cultures were induced from hypocotyl sections of 24 varieties of white clover (Trifolium repens L.). The calli did not show any significant difference of growth among the varieties. After the calli has been transferred to three regeneration media, green-spot formation was observed on calli derived from some seedlings. Remarkable intra- and intervarietal variations in the emergence of green spots and some trends between the origin of varieties and the frequency of green spots were observed. In most cases, the green spots turned brown without showing further differentiation, and only two genotypes formed shoots. A callus from a seedling of the Swedish variety ‘Undrom’ has sustained high levels of plant regeneration throughout 24 months of culture. Protoplasts derived from this selected genotype were divided into cell colonies. 8P (Kao, 1977) medium containing 0.5 mg/1 2,4-D and 0.5 mg/1 kinetin was the most suitable medium for inducing divisions in protoplasts. When subcultured into solid B5 medium, the colonies produced calli, which when transferred to a regeneration medium, formed shoots. This genotype is expected to a useful subject for genetic engineering of white clover.
    Type of Medium: Electronic Resource
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  • 2
    ISSN: 1573-5060
    Keywords: ovule number ; ovule sterility ; seed abortion ; seed set ; Trifolium repens ; white clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A controlled environment study was undertaken to clarify the factors responsible for poor seed set and to study seed development, ovule degeneration and seed abortion, both morphologically and cytologically, in three Japanese cultivars of white clover. Although the mean number of ovules per floret was 4.2–5.1, the average number of seeds per floret was found to be only 2.3–2.7. Microscopic examination of carpels from 0 to 28 days following floret maturity and pollination showed that 26–33% and 8–17% of the total seeds lost occurred within the first three days and the third through fifth day following pollination, respectively. Beyond this period occasional seed abortion was observed at all stages of seed development, but this represented a very small proportion (2–7%) of the total seeds lost. A stain clearing technique was used to examine the cytoplasmic state of the embryo sac in intact, unfertilized, mature ovules and embryos of the ovules at 3 and 5-day periods following pollination. It was found that 20–22% of unfertilized and matured ovules were sterile, suggesting that ovule degeneration before fertilization was the major cause for the high percentage of seeds lost within a 0 to 3-day period following pollination. Cytological observations revealed that abortion of developing seed was due to a sudden arrest in embryo growth and that the early development of the embryo of such aborting seed was normal. Either nutrient shortage or meiotic irregularities may be the cause for high ovule sterility or post-fertilization abortion of developing seeds.
    Type of Medium: Electronic Resource
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  • 3
    ISSN: 1573-5060
    Keywords: Trifolium repens ; white clover ; self-compatibility ; inheritance ; recurrent selection
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary When white clover plants were self-pollinated, 75% of a 143-plant population did not set seed. Plants which did set small numbers of seed probably did so as a result of pseudo-self-compatibility. One highly self-compatible plant was selected from the breeding materials. The S2 progeny of this plant was autogamous and genetically fixed for morphology and isozyme patterns. Genetic analyses of self-compatibility were conducted as far as the F3 progeny. It was concluded from these data that self-compatibility was inherited in a simple Mendelian way and that plants homozygous for self-compatibility were obtained. There was no correlation between presence or absence of self-compatibility and growth vigor. The possibility of application of self-compatible plants to recurrent selection procedures is outlined and the utilization of self-compatible plants in white clover breeding programs is discussed.
    Type of Medium: Electronic Resource
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  • 4
    ISSN: 1573-5060
    Keywords: cryopreservation ; meristemoid ; Trifolium repens ; vitrification ; white clover
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Summary A callus line of white clover capable of forming numerous meristemoids (meristematic cell masses) has been selected and subcultured on agar B5 medium containing 0.5 mg/l 2,4-D and 0.5 mg/l kinetin for three years. The meristematic callus was successfully cryopreserved by vitrification and subsequently regenerated plants. Preculturing callus in liquid B5 medium containing 0.6 M sorbitol at 25°C for 16 hr was essential to the process. Precultured samples (50 mg) were transferred to a 1.8 ml plastic cryotube and then 1 ml of a highly concentrated cryoprotective solution (designated PVS2) was added and mixed. After treatment with PVS2 at 25°C for 7 min or 0°C for 20 min, the sample was directly plunged into LN. After rapid warming, PVS2 was drained from the cryotubes and replaced twice with liquid B5 medium containing 1.2 M sucrose. Samples were transferred onto filter disc over agar B5 medium. Some surviving cells in the cryopreserved meristematic callus proliferated and produced new meristemoids. After 30 days the meristematic callus was transferred onto hormone-free MS agar medium. The meristemoids developed directly into shoots and spontaneously formed roots. Plant regeneration efficiency expressed as a percent of control amounted to about 90%. This vitrification method appears promising as a routine method for cryopreserving meristematic callus of white clover.
    Type of Medium: Electronic Resource
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