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1.
T. Yamada 《Euphytica》1989,44(3):181-186
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.Abbreviations 2,4-D 2,4-dichlorophenoxyacetic acid - IAA indole-3-acetic acid - 2ip 6-, -dimethylallylamino purine - NAA -naphthaleneacetic acid  相似文献   

2.
Embryogenic calli derived from anther cultures of the two-rowed winter barley cultivar Igri were plated on solid L3 medium containing the proline analogue hydroxyproline (Hyp), 10–20 mmol l–1. Exposure to Hyp caused severe degeneration of most of the calli. Hyp resistant calli, distinguishable by their lighter colour and higher growth rate, and control calli not exposed to Hyp were plated on L3 regeneration medium. From 22,500 anthers exposed to Hyp 46 Hyp resistant regenerates were obtained, which were transferred to soil. After cultivation for 5–10 weeks at normal growth conditions they were cold hardened at 2 C under short day conditions together with control regenerates. Frost tolerance assays with segments of fully grown leaves of unhardened and cold hardened plants revealed that Hyp resistant regenerants were significantly more frost tolerant than the control regenerants. Improved frost tolerance was found also in the progenies R1 to R9, and genotypic segregation in the R1 generation in a 1:2:1 ratio was indicated. Increased proline content was observed in the R2 generation and in subsequent generations and was significantly (P 0.001) correlated with increased frost tolerance in the Hyp lines. Comparative studies of R9 progenies from homozygous R2 plants with the wild type Igri under field conditions in winter at three locations in Europe as well as crossing experiments confirmed the heritable improvement of frost tolerance and winter survival, respectively, in the Hyp lines. The results support the hypothesis that proline accumulation in cold acclimated winter barley plants is causally related to the acquisition of frost tolerance. Moreover, the described biotechnological procedure may be applicable in breeding programs for improved winter hardiness and possibly also for other stress tolerances.  相似文献   

3.
A protocol for somatic embryogenesis and plant regeneration of Ilexparaguariensis St. Hil. from embryos cultures was developed. Heart stage zygotic embryos were removed from seeds of immature, light green fruit and treated with antimicrotubule agents (0.1; 0.2, and 0.5% colchicine for 24 and 48 h; 1; 10, and 20 M of either trifluralin, - trifluoro- 2,6-dinitro-N,N- dipropyl-p-toluidine, or oryzalin, 3,5-dinitro-N4, N-dipropylsulphate during 48 h). The embryos were cultured aseptically on quarter-strength Murashige and Skoog medium containing 3% sucrose, 0.65% agar (1/4MS), and 0.46 M zeatin. Cultures were incubated in darkness at 27 ± 2 °C. All thetreatments provoked a diminution of the number of germinated embryos and in some of the treated embryos somatic embryogenesis was induced. Somatic embryo maturation and conversion into whole plants could be achieved by culturing the embryos on 1/4MS lacking hormones and incubated at 27 ± 2 °C, 14 h photoperiod (116 mol m-2s-1). Mostof the plants regenerated from somatic embryos appeared morphologically normaland grew under greenhouse conditions. Only 2 plants out of 152 studied contained the tetraploid number of the chromosomes (2n = 4x = 80), meanwhile the rest of the plants had the normal diploid number of chromosomes (2n =2x = 40). Somatic embryos with abnormal morphology were also observed.  相似文献   

4.
In vitro propagation of Japanese garden iris,Iris ensata Thunb.   总被引：2，自引：0，他引：2  
Summary Explants of young scapes of Iris ensata were cultured on MS medium with 1 mg/l NAA, 1 mg/l 6-BA, 30 g/l sucrose and 10 g/l agar, and this species was characterized by high variety specificity for callus, shoot and root induction. Among 23 varieties and one wild form tested, Okichidori, Miyukisudare and Meiji-l exhibited a considerable rate of shoot induction, although these induced poorly rooted shoots. In addition, two types of callus induction such as green and white calli were observed, and the induction of green-type calli was significantly correlated with that of shoots. Surprisingly, the only modification, half-strength MS inorganic salts, for the above medium proved to be very effective for shoot induction in the scape culture. For shoots obtained from the scape culture, effects of sucrose concentrations and activated charcoal on root induction were examined by using 1/2 MS with 1 mg/l NAA, 1 mg/l 6-BA, 30 g/l sucrose and 10 g/l agar as the basic medium. The addition of 1% activated charcoal to the media had a marked effect for root induction independent of sucrose concentrations and varieties tested. The in vitro propagation technique of I. ensata is discussed.  相似文献   

5.
Summary The effect of anther orientation on culture response has been examined in a range of barley cultivars of agronomic importance. Anther orientation did not significantly affect the percentage of anthers responding. However, orientation of the anthers in the up position with one lobe in contact with the medium significantly increased the number of green plants produced when compared to anthers cultured in the flat position. Green plantlet production was observed directly from embryoids i.e. without an intermediate callus phase. Cytological examination of the regenerants indicated that the majority of the plantlets possessed the normal barley karyotype. The implications of these results for Barley breeding are discussed.  相似文献   

6.
Summary Strawberry cultivars, Redcoat and Veestar, propagated by meristem culture (MC), callus culture (CC) and direct shoot regeneration (DR) from leaf disks were compared for their vegetative and reproductive characters with standard runner (SR) propagated plants under field conditions. In the planting year, in vitro propagated plants of both cultivars had the same number of leaves as SR plants, but in vitro propagated Redcoat produced fewer stolons per plant than SR plants. However, in the following year, in vitro propagated mother plants of both cultivars had more leaves and higher runner production than SR mother plants. Flowering and fruiting behaviour of Veestar was not appreciably influenced by in vitro propagation methods. However, in vitro propagated plants of Redcoat flowered earlier and produced more flowers and fruits than SR plants, but still maintained normal berry weight. Among in vitro propagated plants, DR plants of Redcoat were the earliest to flower, whereas MC plants produced more flowers and fruits. The field performance of the first daughter plants derived from the in vitro propagated plants was consistent with their respective mother plants. Leaf shape of both cultivars was not altered by in vitro propagation. Phenotypic abnormalities were mainly confined to occurrence of yellow leaf variants in MC and CC plants and occasional appearance of plants with irregular flowering and growth habit among CC plants.NRCC No. 38004  相似文献   

7.
In vitro Separation of Chimeral Pears into their Component Genotypes   总被引：2，自引：0，他引：2  
Summary An adventitious shoot regeneration system for Pyrus communis was used to separate two chimeral pears into their component genotypes. The two cultivars were a variegated type, Louise Bonne Panachée and a red fruited mutant, Red Hardy. Leaves of these cultivars were placed onto a regeneration medium consisting of Nitsch & Nitsch (1969) salts supplied with various levels of Thidiazuron (TDZ) and NAA. After two months the regenerants were moved onto a proliferation medium of Lepoivre salts. Later they were evaluated for their chimeral status. Among the regenerants of the variegated type, 100% segregation occurred, most shoots were green, a few were albino. Regeneration was more efficient for dissociating the variegated chimera than rapid shoot multiplication and physical injury. In Red Hardy, after two months on the regeneration medium, 20 to 33% of the regenerants were green, the rest were red. The stability of the red and the green regenerants were assessed on media supplemented with various levels of sucrose and by total anthocyanins measurement. Both types were stable.  相似文献   

8.
R. K. Jain  Ulrike Brune  W. Friedt 《Euphytica》1989,43(1-2):153-163
Summary Protocols for plant regeneration from cotyledon explant and anther cultures of Sinapis alba have been developed for creating doubled-haploids and somaclonal variation. Among the several cultivars tested in this study, only Arda responded well to in vitro plant regeneration both from anther-as well as cotyledoncultures. Multiple shoot formation in cotyledon explants, which always followed a brief callusing phase, was found to be the best on MS medium with ZEA (1.0mg/l) and NAA (0.1mg/l). Regeneration frequency declined sharply in the absence of auxin or presence of other cytokinins and/or auxin. The frequency of shoot regeneration also declined with reduction in the photoperiod to 16h. On MS + BAP (1.0mg/l) + NAA (1.0mg/l) medium, cotyledonary explants showed profuse callusing, which could regenerate shoots on high ZEA + low NAA/IAA medium. However, it declined with progressing time in culture. Anthers, excised from fresh as well as cold pretreated buds, cultured on 10% sucrose containing MS media with different hormonal constitution, developed calli and/or embryos. Initial culture temperature was important with embryogenesis occurring only in anthers cultured at 30°C for 3 weeks. A high temperature (35°C) treatment was lethal for both callus as well as embryo formation. While BAP + NAA and ZEA + NAA/IAA supported embryogenesis, further plant regeneration from anther-or embryo-callus could be achieved in ZEA + NAA/IAA media. Some of the regenerants flowered already in vitro and had small and sterile flowers. Cytological examination of some of the root differentiating calli indicated the presence of haploid as well as diploid cells. Shoots were rooted during prolonged incubation on the same medium or on transfer to MS (reduced)/ B5 + ZEA + NAA media.  相似文献   

9.
Summary Mature anthers of some plants of Narcissus biflorus Curi., cultivated under subtropical conditions at Jammu, were found to contain monads, microspore mother cell embryo sacs, and multinucleate embryoids. In addition, multicellular embryoids resembling those obtained in vitro through anther culture were also observed. All these structures arise from the microspore mother cells which do not follow regular course of microsporogenesis.  相似文献   

10.
S. Tokumasu 《Euphytica》1976,25(1):151-159
Summary Anther development of male-fertile and male-sterile plants in Pelargonium crispum was anatomically examined. Three cultivars, i.e., Lemon crispum, Crispum minor and Prince Rupert, were used. Lemon crispum and Crispum minor are male-fertile, whereas Prince Rupert is male-sterile. The tapetum in every cultivar examined behaved like an amoeba. The tapetal cells of the anther form plasmodial masses. Then, the plasmodial masses fuse producing a periplasmodium. The periplasmodium degenerates and finally disappears. There are no differences in tapetal behaviour between fertile and sterile anthers. In the sterile anthers the endothecium and lip cells do not develop sufficiently. Young microspores show normal growth at early stages. After the completion of their cell wall formation, however, the microspores in the sterile anthers lose their cytoplasm and become empty. On the other hand, the microspores in the fertile anthers increase the volume of their cytoplasm and become fertile pollen grains.  相似文献   

11.
Summary The somatic chromosome numbers of 67 hybrid tuberous begonia cultivars (Begonia x tuberhybrida Voss) have been determined. The material concerned was collected in Belgium and England and represents several distinct groups, comprising both cultivars originated at the turn of this century and those introduced recently.The old Pendula group is characterized by the diploid chromosome number 28. Among the Multiflora group diploid, triploid and tetraploid cultivars are represented almost equally, while the investigated Multiflora maxima and large-flowered begonias proved to be tetraploid except for one triploid.Mitotic instability, to the extent reported by some authors, was not observed. The variation of the diploid number between 27 and 28, the triploid number between 41 and 42 and of the tetraploid number between 52 and 56 can be explained as a consequence of the fact that in the original hybridization wild tuberous species with 2n=26 and 2n=28 were involved.Publication 351, Laboratorium voor Tuinbouwplantenteelt, Landbouwhogeschool, Wageningen, the NetherlandsPublication 23, Rijksstation voor Sierplantenteelt, Melle, Belgium  相似文献   

12.
Summary Thornless Loganberry (TL) is a periclinal chimeral blackberry in which a layer of mutant (thornless) epidermis surrounds a core of wild-type (thorny) tiusse. Due to its chimeral arrangement, TL produces thorny adventitious root cuttings and thorny offspring. To separate the chimera into its components parts, meristems of TL were grow in vitro on modified Murashige and Skoog medium to yield callus and adventitious shoots. One of these shoots has survived, flowered, and produced thornless offspring from seed. The importance of this non-chimeral TL is discussed.List of terms BA 6-benzylaminopurine - GA3 gibberellic acid - NAA naphthalene acetic acid - hybridberry polyploid bramble interspecific hybrids - MS Murashige & Skoog (1962) high mineral salt medium - TL Thornless Loganberry - TLtc tissue culture-derived (non-chimeral) Thornless Loganberry  相似文献   

13.
Summary Somatic embryos of genotype R11 of the alfalfa variety Pampeana were produced from embryogenic calli derived from leaf sections. They were induced by an auxin shock and its development was attempted on six different media. The best condition for somatic embryo production was inducing callus on MS medium plus 10 M 2,4-D and 4,6 M KIN and transferring them, after the auxin shock, to MS with 10–20 mM NH4 + and 30 mM proline. More than 500 somatic embryos per plate were produced. Embryos were grown to plants on MS or half strength MS media and all regenerated plants resembled the original R11 genotype. This technique could be useful in alfalfa Pampeana improvement using genetic modification.  相似文献   

14.
Summary When a local slow bolting variety Osaka Shirona Bansei (Brassica rapa L. ssp. pekinensis, syn. B. campestris L. ssp. pekinensis) was grown in a phytotron (25°C, 16 hours day length without chilling treatment), one third of the plants bolted and flowered. In order to clarify the different flowering responses in the variety, a progeny line (FNC) of the flowering plants was chilled for 4 different periods (0, 22, 36 and 53 days) in a chamber of 2 7°C, then transplanted to three different conditions, i.e. PHY: 25–20°C day and night temperatures, 16 hours day length, GHL: 10 25°C, 16 hours day length with supplementary light and GHN: 10 25°C, natural day length (10 15 hours). In PHY, FNC bolted and flowered with almost the same leaf numbers in all 4 different chilling treatments. This means that FNC has very low sensitivity and no requirement to low temperature for its reproductive growth. In GHN (short day length), FNC bolted very slowly. Then the bolting and flowering of FNC were promoted by both long day length and high temperature. The newly found bolting characteristics of Osaka Shirona Bansei could be applied to breed unique slow bolting Chinese cabbage (B. rapa L. ssp. pekinensis) which might be non-sensitive to low temperature and its bolting and flowering would be induced with the combination of long day length and high temperature. Using the unique variety, it might be also possible to establish a new cropping type of Chinese cabbage (late autumn sowing, spring harvest).Abbreviations PHY Phytotron, long day and high temperature condition - GHL Greenhouse, long day and medium temperature condition - GHN Greenhouse, natural (short) day and medium temperature condition - FNC A progeny line of Osaka Shirona Bansei which flowered with no chilling treatment - FC A progeny line of Osaka Shirona Bansei which flowered with chilling treatment  相似文献   

15.
Summary Germination trials of pollen grains of the tea hybrid rose cultivars: Ferry Porche, Bronze Masterpiece, Queen Elizabeth, John F. Kennedy and Lady X, were carried outin vitro, to determine whether these cultivars are suitable as pollen donors in a breeding programme.Pollen grains of all five cultivars germinated poorly in a medium containing sucrose only. Addition of boric acid in the medium improved the germination percentages. The highest germinability was observed in Ferry Porche and Lady X on 15% sucrose+50 ppm boric acid, and 20% sucrose+100 ppm boric acid, respectively. Addition of calcium nitrate in the medium reduced germination percentages in all cultivars.The effect of pH of the medium on pollen grain germinability depended on the cultivar and the composition of the medium. On a medium free of boric acid, the pH values between 5.5 and 7.0 caused a significant increase in the germination percentages of pollen grains of Lady X and Queen Elizabeth. The addition of boric acid decreased the responsiveness of the pollen of all cultivars to the pH changes of the medium.The germination capacity of pollen grains of Lady X and Ferry Porche was not affected by incubation temperatures ranging between 20°C and 30°C.On the basis of the above results Lady X and Ferry Porche are considered the most suitable for pollen donors.  相似文献   

16.
Some methodological aspects of apple transformation by Agrobacterium   总被引：1，自引：0，他引：1  
Summary Leaf explants of apple cvs Gala and Golden Delicious were infected with the Agrobacterium tumefaciens strain AGL0(pMOG410). The effects of a 2 d preculture of the explants before infection and the addition of 5-azacytidine to the selection medium were studied. The percentages of GUS-positive explants after 5 w did not significantly alter due to these treatments. One of the Gala shoots, which was removed from a leaf explant cultured for 8 w on selection medium, proved to be GUS-positive and will be analyzed further. In general, however, it should be concluded that regeneration of transgenic shoots directly from leaf tissue was not very effective.Abbreviations MS Murashige and Skoog medium - GD Golden Delicious  相似文献   

17.
Summary Nuclear and cytoplasmic factors affect tissue culture response in wheat (Triticum aestivum), and cytoplasmic male sterility may enhance callus initiation in anther culture. Three wheat nuclear genotypes, each in normal and two alien cytoplasms conferring cytoplasmic male sterility, were evaluated for callus initiation frequency in anther culture. Nuclear genotype had the greatest effect on callus initiation, but cytoplasm and nucleus X cytoplasm interaction also produced significant effects. The nuclear genotype of Chris outperformed Butte and Coteau in all cytoplasms. Ordinary wheat and Triticum timopheevi cytoplasms outperformed Aegilops speltoides cytoplasm. Cytoplasmic male sterility did not increase callus initiation. This suggests manipulating the nuclear genotype is the best strategy for improving the capacity of wheat to initiate callus in anther culture.  相似文献   

18.
M. Sujatha 《Euphytica》1997,93(1):89-95
A rapid and efficient method of in vitro plant regeneration for large scale propagation of male sterile plants of niger (Guizotia abyssinica) was developed. Leaf segments from mature plants were cultured on Murashige & Skoog's basal medium (MS) supplemented with N6-benzyladenine (BA) and kinetin individually and in combination with low concentrations of indole-3-butyric acid (IBA); and -Naphthaleneacetic acid (NAA). Prolific direct adventitious shoot regeneration occurred on most of the media tested. The best response in terms of frequency of shoot regeneration and the number of shoots per leaf explant was observed on medium supplemented with 2.22 M BA. Transfer of shoot bud clusters to fresh medium with same composition promoted further multiplication of the shoot buds, while medium with reduced BA concentration (0.89 M) facilitated shoot elongation. Shoots that were rooted on half-strength MS medium gelled with 0.2 or 0.4% agar and supplemented with 4.9 M IBA survived with a frequency of 61.36% on transfer to ex vitrum conditions. Field evaluation of the regenerants revealed the genetic stability of the plantlets and are being used in breeding of experimental hybrids.  相似文献   

19.
Summary The determinate Phaseolus vulgaris L. cvs Dark Red Kidney Charlevoix, and near-isogenic Great Northern Nebraska 1 were grown in blends with the indeterminate GN Nebraska 1 in white mold (Sclerotinia sclerotiorum) disease field nurseries. A critical difference between Charlevoix and indeterminate GN Nebraska 1 is that the latter has a greater leaf area closer to the soil surface, and this is associated with increased numbers of apothecia beneath the canopy and higher disease severity. White mold infection and apothecia number/m2 beneath the canopy of the blends containing Charlevoix were significantly reduced in comparison with the severely infected, homogeneous, indeterminate GN Nebraska 1. A reduction of white mold infection for the indeterminate GN Nebraska 1 was not observed in blends grown under severe or moderate white mold incidence, but did occur under slight incidence in blends containing 65% and 75% Charlevoix.No significant difference for seed yield occurred between the blends and homogeneous cultivars planted in four experiments under severe, moderate, slight and zero white mold incidence, respectively, except in Experiment 1 under moderate white mold incidence, the blend of 50% indeterminate GN Nebraska 1 and 50% Charlevoix exceeded the yield of the indeterminate GN Nebraska 1. Mean weight of white (GN Nebraska 1) and red seed (Charlevoix) increased and decreased, respectively, in some blends due to the more vigorous growth of the indeterminate GN Nebraska 1.Published as Paper No. 5341, Journal Series, Nebraska Agric. Exp.t Station. Research was conducted un under Project 20–3.  相似文献   

20.
Summary The investigation involved three barley genotypes that varied from extremely susceptible (Akka) to an extreme level of partial resistance (17-6-16). The barley leaf rust colony size was measured in primary leaves 3, 6, 12 and 18 days after inoculation and in flag leaves 6, 12, 18, 24 and 30 days after inoculation with race 1-2-1. Akka always had the largest colonies, 17-5-16 the smallest, with Vada at an intermediate position. The genotypic differences were proportionally largest at the second sampling day and smallest at the last sampling day. The rate of colony growth decreased rapidly over time for all genotypes and in both plant stages. Measured at the same time (colony age the same) the rate of colony growth was largest for 17-5-16 and smallest for Akka in most periods. The time needed to reach a given colony size showed already large differences in the very early states especially in the flag leaves. Akka took 3.9 days to reach a size of 17 × 10-3 mm2 (only 5 to 10% of the colony size at the start of sporulation), Vada needed 8.3 days and 17-5-16 even 12.0 days. To reach a size of 320 × 10-3 mm2 the three genotypes needed 12.7, 18.0 and 22.8 days respectively, differences that are only slightly larger than those at the very small colony size.It was concluded that the partial resistance of barley to barley leaf rust is not primarily due to a reduced fungal growth in the partially resistant host tissue but predominantly so to an initial and temporary stagnation at the site of penetration. The longer this stagnation lasts, the longer the latent period and the higher the partial resistance are. Once this stagnation has been overcome the fungal growth rates do not vary much between genotypes with different levels of partial resistance.  相似文献   

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