共查询到20条相似文献，搜索用时 410 毫秒
1.
Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks. 相似文献
2.
G. Spangenberg Z. Y. Wang G. Legris P. Montavon T. Takamizo R. Pérez-Vicente M. P. Vallés J. Nagel I. Potrykus 《Euphytica》1955,85(1-3):235-245
Summary Intergeneric symmetric and asymmetric somatic hybrids have been obtained by fusion of metabolically inactivated protoplasts from embryogenic suspension cultures of tall fescue (Festuca arundinacea Schreb.) and unirradiated or 10–500 Gy-irradiated protoplasts from non-morphogenic cell suspensions of Italian ryegrass (Lolium multiflorum Lam.). Genotypically and phenotypically different somatic hybrid Festulolium mature flowering plants were regenerated.Species-specific sequences from F. arundinacea and L. multiflorum being dispersed and evenly-represented in the corresponding genomes were isolated and used for the molecular characterization of the nuclear make-up of the intergeneric, somatic Festulolium plants recovered. The irradiation of Italian ryegrass protoplasts with 250 Gy X-rays prior to fusogenic treatment favoured the unidirectional elimination of most or part of the donor chromosomes. Irradiation of L. multiflorum protoplasts with 500 Gy produced highly asymmetric (over 80% donor genome elimination) nuclear hybrids and clones showing a complete loss of donor chromosomes.The RFLP analysis of the organellar composition in symmetric and asymmetric tall fescue (+) Italian ryegrass regenerants confirmed their somatic hybrid character and revealed a bias towards recipient-type organelles when extensive donor nuclear genome elimination had occurred.Approaches aimed at improving persistence of ryegrasses based on asymmetric somatic hybridization with largely sexually-incompatible grass species (F. rubra and Alopecurus pratensis), and at transferring the cytoplasmic male sterility trait by intra- and inter-specific hybridization in L. multiflorum and L. perenne, have been undertaken.Abbreviations cpDNA
chloroplast DNA
- CMS
cytoplasmic male sterility
- 2,4-D
2,4-dichlorophenoxy-acetic acid
- IOA
iodoacetamide
- mtDNA
mitochondrial DNA
- RFLP
restriction fragment length polymorphism 相似文献
3.
Protoplasts from cell suspension cultures of ‘Bonnaza’ navel orange (Citrus sinensis (L.) Osbeck) were electrically fused with mesophyll protoplasts isolated from seedless ‘Red Blush’ grapefruit (Citrusparadisi). After 6 months of culture, a total of 20 plants were regenerated. Root tip chromosome counting revealed that 4 of them
were tetraploids (2n = 4x = 36)and the rest were diploids (2n = 2x = 18) morphologically resembling the mesophyll parent.
After 6 months of transplantation into the greenhouse, 4 of the diploidmesophyll regenerants unexpectedly flowered, but this
phenomenon disappeared in the next year. This is the first report of precocious flowering in citrus via protoplast fusion.
RAPD analysis further confirmed that the tetraploid regenerants were somatic hybrids while the diploid regenerants were mesophyll
parent type. This somatic hybrid will be utilized as a possible pollen parent for improving the seedy pummelo cultivars in
China by producing triploid seedless pummelo hybrid. The mechanism of early flowering was also discussed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
4.
V. Orbović M. Ćalović Z. Viloria B. Nielsen F. G. Gmitter Jr. W. S. Castle J. W. Grosser 《Euphytica》2008,161(3):329-335
Five populations of lemon plants [Citrus limon (L.) Burm] obtained from undeveloped ovules through different tissue culture procedures were examined for the presence of
somaclonal and irradiation-induced genetic variation. Tested groups were: (1) nucellar seedlings; (2) organogenic, regenerated
via adventitious buds from nucellar seedling internodes; (3) embryogenic population, regenerated from non-irradiated nucellar
callus via somatic embryogenesis; (4) embryogenic population, regenerated from irradiated nucellar callus via somatic embryogenesis;
and (5) protoplast-derived, regenerated via somatic embryogenesis. Genomic DNA samples from 360 plants (72 from each group)
were screened for polymorphism among RAPD fingerprints amplified by 10 decamer primers. Among all tested plants, genetic variation
was detected only within the group of plants recovered from irradiated embryogenic calli. Out of 72 plants from that group,
three had RAPD fingerprints different from the rest of the population, and fourth plant was found to be cytochimeric, consisting
of diploid and tetraploid cells as revealed by flow cytometry. In all other populations of regenerated plants, we did not
come across any plants with changed ploidy level. 相似文献
5.
Veli-Matti Rokka Yong-Sheng Xu Jyri Kankila Anja Kuusela Seppo Pulli Eija Pehu 《Euphytica》1994,80(3):207-217
Summary Symmetric somatic hybrids were produced by electrofusion of protoplasts of two dihaploid tuber-bearing potato (Solanum tuberosum L.) lines and Solanum brevidens Phil., a diploid non-tuber-bearing wild potato species. A total of 985 plants was obtained. Verification of nuclear hybridity of putative hybrids was based on additive RAPD patterns, general morphological characteristics and chromosome counts. 53 (90%) calli regenerated into plants which were identified as somatic hybrids. Most of the hybrids were aneuploids at the tetraploid (4×) or hexaploid (6×) level. The 20 hybrids tested expressed a high level of resistance to potato virus Y (PVY
N
) characteristic of the S. brevidens parent. Resistance to late blight (Phytophthora infestans (Mont.) de Bary) varied between hybrids, but was on average better than that of the fusion parents. Resistance of hybrids to bacterial stem rot (Erwinia carotovora subsp. atroseptica (van Hall) Dye) was not superior to that of commercial potato cultivars. 相似文献
6.
Seed dormancy mechanisms in warm season grass species 总被引：6，自引：0，他引：6
Embryogenic protoplasts of Dancy tangerine (Citrus reticulata Blanco) were X-ray irradiated at three doses and electrofused with iodoacetic acid-treated embryogenic protoplasts of Page
tangelo (C. reticulata Blanco × C. paradisi Macf.). Shoots could regenerate only from the fusion combination with the lowest irradiation dose, but were recalcitrant
to rooting. In vitro grafting was applied to obtain complete plants. Chromosome examination showed that the plants contained mainly diploid and
aneuploid cells, together with few tetraploid cells, indicating that they were mixoploids. Random amplified polymorphic DNA
analyses with 10-mer arbitrary primers confirmed the plants as true somatic hybrids. This is the first report on regeneration
of mixoploid hybrid plants via protoplast asymmetric fusion in Citrus. Negative effects of ionizing irradiation on regeneration of embryoids and plantlets and possible agronomic interest of the
mixoploid plants are also discussed.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
7.
Somatic hybridization has been an effective and successful technique for plant improvement. In this paper, embryogenic callus protoplasts of red tangerine (Citrus reticulata Blanco) were electrically fused with mesophyll protoplasts from citrange (C. sinensis × P. trifoliata, a Chinese local strain) in an effort to produce complementary tetraploid citrus rootstocks. Regenerated embryoids grew slowly and were vulnerable to browning. Twelve plants were finally regenerated, rooted and transplanted into a greenhouse. Root‐tip chromosome counting of five randomly‐selected plants revealed most cells were tetraploid (2n = 4x = 36), but aneuploid cells also existed. Flow cytometry analysis further confirmed their tetraploid nature. Nuclear simple sequence repeat (SSR) analysis verified their hybridity. Further mitochondrial genome analysis by restriction fragment length polymorphism and cleaved amplified polymorphic sequence revealed their mtDNA banding pattern was identical to that of red tangerine, the embryogenic callus parent; while their chloroplast DNA inheritance was random as revealed by chloroplast SSR analysis, in addition to cpDNA co‐existence detected in one plant. Cytological and molecular analysis indicated that somatic hybrid plants between red tangerine and citrange had been successfully obtained. 相似文献
8.
The present investigation discusses the scope for transferring of resistance to leaf spot disease incited by Alternaria helianthi from two hexaploid wild species (H. tuberosus and H. resinosus) to diploid cultivated sunflower. Interspecific hybrids produced between sunflower and these two hexaploid species were partially fertile with tetraploid chromosome status. Backcrosses of these interspecific hybrids with cultivated sunflower resulted in the formation of sterile triploid plants. To overcome the problem of sterility and facilitate backcrosses with cultivated sunflower, anther culture of the tetraploid interspecific hybrids was carried out to bring down their chromosome number to diploid status. Anthers from both interspecific hybrids were cultured on basal Murashige and Skoog media supplemented with varying concentrations of organics and the growth regulators benzyladenine and naphthaleneacetic acid. Anthers of interspecific hybrids involving H. resinosus responded well and regenerated through an embryogenic route at a frequency of 98.7%. But in interspecific hybrids with H. tuberosus, anthers formed callus and subsequently regenerated shoots through an organogenic pathway. DNA ploidy analysis of anther culture plants of interspecific hybrids derived from H. tuberosus crosses was carried out to identify plants with desired diploid status. In vitro screening of parents, interspecific hybrids and anther culture plantlets against A. helianthi showed resistance in 68.5% of the anther culture plants of interspecific hybrids from H. tuberosus and in 24.3% of the plants derived from interspecific hybrids involving H. resinosus. 相似文献
9.
Of 3272 plants regenerated from protoplasts of 10 Saintpaulia ionantha genotypes, 98.4% survived transfer to the greenhouse. The frequency of regenerants with chlorophyll deficiencies, i.e. variegated leaves or albinos, was low (1.5%). There was a higher number of polyploid, in most cases tetraploid plants, regenerated from protoplasts (16%) which were identified by their altered morphology. Measurements of stomatal length and counting the number of chloroplasts per guard cell also allowed a clear differentiation between diploid and polyploid plants. The classification was confirmed by DNA content determination using flow cytometry. Mechanisms leading to polyploidization included spontaneous protoplast fusion as well as chromosome doubling during callus growth and shoot regeneration. Two genotypes with instabilities in flower colour showed completely altered flower colours in plants regenerated from protoplasts as well as in plants regenerated on leaf explants in vitro. 相似文献
10.
Plant regeneration from protoplasts of Iris germanica L. 总被引：1，自引：0，他引：1
Summary Protoplasts were isolated enzymatically from suspension cultures derived from embryogenic calli induced by leaf base culture of Iris germanica. In protoplast culture, the effects of glucose concentration, different sugars and combinations of 2,4-D and KIN on protoplast division and colony formation were examined. N6 medium supplemented with 0.1–1 mg/l 2,4-D, 1 mg/l KIN, 200mg/l casein hydrolysate, 250 mg/l proline, 0.2 M glucose and 20 g/l agarose was suitable for protoplast division and colony formation. When colonies formed were transferred onto hormone-free MS medium, many plantlets were regenerated through somatic embryogenesis. Thus, we could establish a plant regeneration system from protoplasts of I. germanica.Contribution from the Laboratory of Plant Breeding, Faculty of Agriculture, Miyazaki University, Japan, No. 95. 相似文献
11.
M. Koornneef J. A. M. van Diepen C. J. Hanhart A. C. Kieboom-de Waart L. Martinelli H. C. H. Schoenmakers J. Wijbrandi 《Euphytica》1989,43(1-2):179-186
Summary The effect of the tissue culture system, the genotype and the ploidy level of the plant material used as explant source on the stability of the ploidy level of plants regenerated fromcell and tissue cultures of tomato was investigated. In addition the use of the chloroplast number in guard cells as a measure for ploidy level was evaluated. Haploids of tomato were very instable, which instability was observed already in somatic root tip and leaf cells. The number of regenerated plants that retained the original ploidy level differed significantly between the tested haploids. The plants that were regenerated from leaf explants of diploids were predominantly diploid in contrast to the plants regenerated from established callus cultures and protoplast where the majority was tetraploid. 相似文献
12.
Summary Interspecific somatic hybrid plants were regenerated after electrofusion of mesophyll protoplasts with the objective of transferring resistance to Verticillium dahliae from Solanum torvum into potato. Early selection of the putative hybrids was based on differences in cultural behaviour of the parental and hybrid calli (particularly the ability of the latter to regenerate early) in combination with morphological markers. Four putative hybrids were recovered from hundreds of calli, probably resulting from complementation of the two parental genomes. The regenerates were tetraploids (2n=4×=48 chromosomes) and exhibited intermediate traits including leaf form, plant morphology and the presence of anthocyanin. The hybrid nature of the four selected plants was confirmed by examining isoenzyme patterns for isocitrate dehydrogenase (Idh), malate dehydrogenase (Mdh), phosphoglucoisomerase (Pgi) and 6-phosphogluconate dehydrogenase (6-Pgd). While the hybrid plants rooted readily and grew vigorously under in vitro conditions, in the greenhouse their development and growth were retarded by difficulties in rooting. When grafted on potato or S. torvum rootstocks, the hybrid plants recovered normal development and growth. Again, they exhibited intermediate morphological traits. Tests for resistance realized in vitro with medium containing 50% Verticillium wilt filtrate showed that all the somatic hybrids were resistant to the fungus filtrate. 相似文献
13.
The induction of tetraploidy in chicory (Cichorium intybus L. var. Magdebourg) by protoplast fusion 总被引：2，自引：0，他引：2
Summary Fusions of chicory leaf protoplasts were realized in order to obtain a higher number of tetraploid plants compared to the traditional doubling techniques with colchicine. Thanks to the fusion technique, using a mixture of PEG, DMSO and a solution of CaCl2 (pH 10.5), 25% tetraploid (2n=36) plants were obtained. Among the 167 regenerated plants, only one aneuploid of a near tetraploid level (2n=33) was identified. 相似文献
14.
Summary This article reports the culture and plant regeneration of Tripsacum dactyloides. Mature embryos of Tripsacum dactyloides dactyloides were used to obtain embryogenic callus cultures. Currently, 180 normal plants have been regenerated from these cultures. Callus was initiated on MS medium supplemented with dicamba (10 mol or 20 mol) and sucrose (3% or 6%), and plants were regenerated on hormone free MS medium containing 2% sucrose. No significant differences were found in callus initiation frequency or in embryogenic response of cultures on the four combinations of sucrose and dicamba tested. The embryogenic cultures have been maintained for 9 months (12 subcultures) and have retained regeneration capacity. Plants regenerated from tissue culture of maize-by-Tripsacum hybrids could be useful in maize improvement. 相似文献
15.
Summary Somaclonal variation, which is a welcome source of genetic variation for crop breeding, is unwanted when direct regenerants have to be used in tissue culture mass propagation (eg. in many forest trees), or in the regeneration of genetically transformed plants. Random amplified polymorphic DNA (RAPD) was used to analyse somatic embryos and plants regenerated from embryogenic cell lines in Norway spruce, Picea abies (L.) Karst. RAPD facilitated the identification of clones, as material from the same cell lines shared identical patterns of amplified fragments, whereas regenerants from different cell lines were easily distinguishable by their respective patterns. For comparisons with explant donor genotypes, cell lines were initiated from cotyledons. Some of the seedlings that had parts of their cotyledons removed were grown on as control plants. Somatic embryos regenerated from cotyledon cell lines showed no aberrations in RAPD banding patterns with respect to donor plants. We conclude that gross somaclonal variation is absent in our plant regeneration system.Abbreviations ESM
embryogenic suspensor mass
- RAPD
random amplified polymorphic DNA
- RFLP
restriction fragment length polymorphism
- (2,4-dichlorophenoxy)acetic acid
2,4-D
- 1-naphthaleneacetic acid
NAA 相似文献
16.
Summary Experimental conditions were established that resulted in high yields and good viability of the protoplasts obtained from leaves of Vitis vinifera, cv. Chardonnay regenerated in vitro by somatic embryogenesis. The effect of factors of the culture medium and various environmental conditions upon the frequency of cell division has been examined, and a method of culture is described by which protoplasts were induced to begin division. Most protoplasts obtained in this way regenerated cell walls within the first few days and cell division occurred after 10 days of culture in a liquid medium. Some cells have divided two or even three times. Nevertheless, the cells did not continue dividing beyond this stage. 相似文献
17.
Somatic hybridization can be used to induce genetic variability in plastidial and mitochondrial genomes, and transfer multiple
uncloned genes across sexual barriers. Somatic hybrids were produced between a dihaploid clone of the common potato, S. tuberosum subsp. tuberosum, and the wild sexually incongruent diploid species S. commersonii. Fusion products were selected on the basis of callus growth and regeneration in vitro. Genome composition of putative somatic hybrids was determined by flow cytometric analysis of nuclear DNA content, RAPD analysis,
and Southern analysis with probes specific to organellar DNA. All regenerated fusion products proved to be hybrids based on
RAPD analysis. Seventy per cent of somatic hybrids were (near) tetraploids, 22% (near) hexaploids and 8%(near) octoploids.
A high correlation was found between the nuclear DNA content determined by flow cytometry and the number of chloroplasts in
stomata guard cell pairs. Somatic hybrids inherited the parental plastids in a random manner. On the contrary, they preferentially
inherited the mitochondrial DNA fragments of S. tuberosum. The majority of them had a rearranged mitochondrial genome with fragments from both parents. Hybrids were highly vigorous
and morphologically more similar to the cultivated than to the wild parent, produced tubers on long stolons under long photoperiod
conditions, showed a high degree of flowering, but did not produce pollen. In addition, somatic hybrids were generally more
resistant to frost and Verticillium wilt than the cultivated parent, indicating the introgression of relevant resistance genes
from the wild species into the genetic background of S. tuberosum.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
Growth and yield characteristics of two different clones of banana plants (Musa AAA cv. Grande naine) originating from four months old embryogenic cell suspensions were studied. These characteristics were
compared with those plants produced by the conventional in vitro budding multiplication method. Two types of variants were observed during the acclimatization phase among 500 embryogenic
cell suspension derived plants. The first type related to banana plants with `variegated or deformed leaves' were also observed
in in vitro budding derived plants. The second type concerned `fasciated-leafed' plants. During the field growth, these two variant types
produced plants morphologically similar to the other plants. Thus, none of the cell suspension derived plants exhibited off-type
traits in the field. A Fisher block model was used to compare the field performances of the two clones produced through the
two in vitro propagation techniques. The analysis of variance showed that there were no significant differences between the plants produced
by either micropropagation techniques for the plant height and circumference, the length of the reference leaf, the number
of nodal clusters of the inflorescence and of fruits, the bunch weight, the period of time between planting and flowering,
and between planting and harvesting. This study showed that banana plants with an agronomical behaviour similar to those produced
by the conventional in vitro budding method could be regenerated from embryogenic cell suspension.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
19.
María Valeria Romagnoli Juan Pablo A. Ortiz Gerardo D. Cervigni Cintia Heisterborg Rubén H. Vallejos 《Euphytica》1996,90(1):89-93
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
20.
芥菜型油菜和黑芥与诸葛菜属间杂种的获得及其特性 总被引：2，自引：0，他引：2
属间远缘杂交是创造新种质、解决育种材料短缺的一种有效方法。为了使诸葛菜的优良基因转入芥菜型油菜和黑芥中, 采用子房培养技术, 进行芥菜型油菜和黑芥与诸葛菜的正反属间杂交。在6种不同的培养基中培养子房。结果表明, 不同的杂交组合、培养基和母本的属间杂交可交配性不同。通过子房培养, 获得了10株属间杂种, 其中9株在形态上近似母本, 或介于父母本之间, 尤嫩斯与诸葛菜杂交组合的1株杂种表现出完全的雄性不育, 并在分枝类型和叶型方面表现出明显的变异; 形态学和SSR分子生物学鉴定表明10株杂种都是真正的属间杂种。 相似文献