共查询到20条相似文献,搜索用时 62 毫秒
1.
M. Q. Wang Z. Y. Peng M. C. Wang W. Guo J. S. Zhao D. Y. Zhi G. M. Xia 《Russian Journal of Plant Physiology》2011,58(4):615-621
Protoplasts of Bupleurum scorzonerifolium irradiated with 380 μW/cm2 UV for 5 min were fused by the PEG-mediated method with untreated protoplasts of Arabidopsis thaliana. The fusion products were cultured in the P5 liquid medium for single hybrid cell clone formation. As a total, 81 independent
putative hybrid clones (cell lines) were obtained, and seventeen of them were identified as somatic hybrids by chromosome
counting, GISH, RAPD, and SSR analyses. More than 80 B. scorzonerifolium-like green introgressed plants and leaves were regenerated from 49 somatic hybrid cell lines, which contained chromatin and
DNA characteristic of A. thaliana. To assess the UV tolerance of both parents with chromatin exclusion and introgression, their protoplasts were UV-irradiated
(380 μW/cm2 for 0 and 5 min), and the protoplasts of A. thaliana were more sensitive to UV than those of B. scorzonerifolium as judged by Single Cell Gel Electrophoresis analysis. The possible relationship between UV resistance of B. scorzonerifolium and A. thaliana chromosome elimination and the formation of somatic introgressed hybrid plants is discussed. 相似文献
2.
M. Wang Z. Peng L. Wang J. Zhao J. Che G. Xia 《Russian Journal of Plant Physiology》2011,58(1):133-141
The protoplasts of tall fescue (Festuca arundinacea Schreb.) were fused with those of Bupleurum scorzonerifolium Willd. The latter were irradiated with UV at an intensity of 380 μW/cm2 for 0 s (combination I), 30 s (combination II), and 60 s (combination III) before fusion. Putative hybrid calli, leaves,
and shoots were generated from the fusion products. They were recognized as somatic hybrids by a combined analysis of chromosome
numbers, isozyme, RAPD, and 5S rDNA spacer sequence. The hybrid calli with morphogenetic ability and leaves/shoots differentiation
had the B. scorzonerifolium phenotype, whether they were derived from symmetric fusion (UV 0 s) or asymmetric fusion (UV 30 s/60 s). Cytological tests
revealed that these hybrids contained the complete set (12) of B. scorzonerifolium chromosomes and 0–4 partner tall fescue chromosomes. The tall fescue chromosomes were rapidly eliminated in combinations
II and III, but gradually lost in combination I. It was noted that the green leaves and shoots were produced earlier, and
the differentiation frequency was higher in combinations II and III than in combination I, which corresponded to the speed
of elimination of the tall fescue chromosomes in the hybrids. Therefore, UV irradiation can indirectly promote elimination
of tall fescue chromosomes and hybrid differentiation. B. scorzonerifolium can repel partner chromosomes with mechanism that differs from UV. 相似文献
3.
In this paper, we describe how Bupleurum scorzonerifolium/Triticum aestivum asymmetric somatic hybrids can be exploited to study the wheat genome. Protoplasts of B. scorzonerifolium Willd were irradiated with ultraviolet light (UV) and fused with protoplasts of common wheat (T. aestivum L.). All cell clones were similar in appearance to those of B. scorzonerifolium, while the regenerated plantlets were either intermediate or B. scorzonerifolium-like. Genotypic screening using isozymes showed that 39.3% of cell clones formed were hybrid. Some of the hybrid cell clones grew vigorously, and differentiated green leaves, shoots or plantlets. DNA marker analysis of the hybrids demonstrated that wheat DNA was integrated into the nuclear genomes of B. scorzonerifolium and in situ karyotyping cells revealed that a few wheat chromosome fragments had been introgressed into B. scorzonerifolium. The average wheat SSR retention frequency of the RH panel was 20.50%, but was only 6.67% in fusions with a non-irradiated donor. B. scorzonerifolium chromosomes and wheat SSR fragments in most asymmetric hybrid cell lines remained stable over a period of 2.5–3.5 years. We suggest the UV-induced asymmetric somatic hybrids between B. scorzonerifolium Willd and T. aestivum L. have the potential for use in the construction of an RH map of the wheat genome. 相似文献
4.
Wang Minqin Zhao Junsheng Peng Zhenying Guo Wei Wang Yun Wang Le Xia Guangmin 《Plant Cell, Tissue and Organ Culture》2008,92(2):121-130
In order to investigate chromosome elimination in symmetric somatic hybridization between Bupleurum scorzonerifolium and Arabidopsis thaliana, protoplasts were isolated from suspension cultures of both A. thaliana and B. scorzonerifolium parents. Biparental protoplasts were mixed at a rate of 1.5:1 and fused with PEG-method. After protoplast fusion, the products
were cultured in the P5 liquid medium for microcallus formation. Single cell lines formed from microcalli after subculturing
on the MB1 (Xia and Chen, Plant Sci 120:197–203, 1996) solid medium. The putative somatic hybrid cell lines were identified
by cytological and molecular analysis. Of the 132 somatic cell lines generated, 16 were identified as somatic hybrids, with
the phenotypes resembled B. scorzonerifolium parent. These hybrids showed a complete set of B. scorzonerifolium chromosome and 0–2 small chromosome(s) of A. thaliana. A few of them showed nuclear and cytoplasmic SSR fragments of A. thaliana. These hybrid cell lines could differentiate to green spots, buds/leaves through complementation of regeneration ability.
The chromosomes elimination of A. thaliana was discussed.
Wang Minqin and Zhao Junsheng contributed equally to the work. 相似文献
5.
In our early experiments, a variety of Bupleurum scorzonerifolium-like somatic hybrid plants were obtained from protoplast fusion between Arabidopsis thaliana and UV-treated/untreated B. scorzonerifolium. To compare the effects of UV and γ-ray irradiation on the B. scorzonerifolium partner and obtain Arabidopsis-like hybrids, we designed a novel combination of somatic hybridization between A. thaliana and B. scorzonerifolium. Before protoplast isolation and fusion, the suspension cells of B. scorzonerifolium were irradiated by gamma ray (60Co, 50 Gy with 1.3 Gy min−1). Both parental protoplasts lost regeneration capacity, but over 100 somatic hybrids restored the capacity and developed
to Arabidopsis-like inflorescences and flowers with some characteristics of B. scorzonerifolium. Some hybrid flowers showed yellow sepal, petal, or carpel, whose color was similar to the petal of B. scorzonerifolium; the others had silique of Arabidopsis with angularity of B. scorzonerifolium, and their parts possessed five stamens, the same as B. scorzonerifolium. Cytological analysis showed that three hybrids had Arabidopsis-like karyotypes. Random Amplified Polymorphic DNA (RAPD) and Simple Sequence Repeats (SSR) profiles revealed that both parental
fragments were amplified from these hybrids. These results indicated chromatin introgression from B. scorzonerifolium to A. thaliana, which may be related to the complementation of hybrid inflorescence and flower generation. 相似文献
6.
T. Cardi E. D. Earle 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,94(2):204-212
New types of cytoplasmic male sterility (CMS) in Brassica oleracea would be useful for F1 hybrid seed production. The `Anand' cytoplasm derives from the wild species B. tournefortii. Rapid cycling stocks of B. rapa and B. oleracea were used in cybridization experiments as donor and recipient of `Anand' (=`tour') CMS, respectively. Prior to fusion with
PEG, donor protoplasts were inactivated with 30 krad γ-rays and recipient ones with 3 mM iodoacetate, respectively. No calli were obtained from the pre-treated protoplasts. The frequency of shoot regeneration was
21–43% in untreated B. oleracea controls, but only 0–0.5% in `Anand' B. rapa. Putative cybrids were regenerated from about 3% of the calli from fused protoplasts. Regenerated plants were analyzed for
nuclear DNA content, plant and flower morphology, pollen production, female fertility, cold tolerance, and organelle composition.
Eighty-one percent of the regenerated controls and 63% of fusion-derived plants were diploid. The rest showed DNA contents
corresponding to 2x–4x, 4x, or higher ploidy levels, presumably due to somatic doubling in vitro and/or fusions in which the donor nucleus was not completely
eliminated. Sixty-four percent of the cybrids had stamens and petals varying in size and shape and were male-sterile, with
indehiscent anthers. Their phenotype was otherwise similar to that of B. oleracea. The remaining plants had normal flowers and were male-fertile. Data from crosses with fertile pollinators indicated good
female fertility in some of the sterile lines, both after hand and insect pollinations in cages. Mitochondrial (mt) segregation
in the cybrids was slightly biased towards `Anand' mitochondria, and the presence of `Anand' mtDNA fragments was strongly
associated with male sterility. Evidence of mtDNA rearrangements was obtained in some cybrids. Segregation of chloroplasts
was slightly biased towards B. oleracea. The presence of `Anand' chloroplasts with a B. oleracea nucleus did not result in cold temperature chlorosis, as seen in `Ogura' CMS plants.
Received: 22 February 1996 / Accepted: 10 May 1996 相似文献
7.
The use of interesting characteristics from wild Helianthus species in sunflower breeding is limited by poor crossability or sterility of interspecific hybrids. To overcome this barrier,
mesophyll protoplasts of Sclerotinia sclerotiorum-resistant clones of Helianthus maximiliani, H. giganteus and H. nuttallii were fused with hypocotyl protoplasts of H. annuus in the presence of polyethyleneglycol and dimethylsulfoxide. Fusion products were embedded in agarose and subjected to a
regeneration protocol developed for sunflower protoplasts. Organogenic calli were transferred onto solid medium and emerging
shoots were elongated in the absence of plant growth regulators. Rooting of shoots was induced by a 1-naphthaleneacetic acid
treatment and putative hybrid plants from fusions between H. annuus + H. maximiliani and H. annuus + H. giganteus were transferred into the greenhouse. All of them exhibited a hybrid phenotype with a high percentage of rhizome producing
plants. Their hybrid origin was confirmed by random amplified polymorphic DNA analysis. Plants flowered after 3–4 months and
set seeds, of which 70–80% germinated.
Received: 18 February 1997 / Revision received: 25 March 1997 / Accepted: 15 May 1997 相似文献
8.
J. O. Rasmussen S. Waara O. S. Rasmussen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1997,95(1-2):41-49
Recipient protoplasts from three Solanum tuberosum genotypes, cv ‘Folva’ (2n=4x=48), cv ‘Matilda’ (4n) and ‘161 : 14’ (2n), were electrofused with X-ray-irradiated donor protoplasts
from two wild species S. spegazzinii (2n) or S. microdontum×S. vernei (2n). Prior to fusion, protoplasts were fluorescence-labelled with either fluorescein diacetate or scopoletin. Fusion products
were identified by dual fluorescence and selected by micromanipulation or fluorescence-activated cell sorting (FACS). All
putative hybrid plants were analysed by the random amplified polymorphic DNA (RAPD) technique. Our analysis demonstrates that
each asymmetric hybrid plant has an individual and stable profile of donor-specific RAPD bands. The irradiation of donor protoplasts
hampered the growth of selected heterofusion products in a dose-dependent way. Irradiation resulted in donor chromosome elimination,
but not in a dose dependent way, in the tested interval. In asymmetric hybrids with the S. spegazzinii donor 33–68% of the donor-specific RAPD bands were missing, indicating a similar level of chromosome elimination. In asymmetric
hybrid plants with the S. microdontum×S. vernei donor 74–95% of the donor RAPD bands were missing. Chromosome countings revealed that these hybrids had chromosome numbers
equal to or below the chromosome numbers found in the tetraploid recipients. This is the first time that highly asymmetric
hybrid plants between two tetraploid potato recipients and the donor S. microdontum×S. vernei have been obtained.
Received : 16 December 1996 / Accepted: 21 February 1997 相似文献
9.
Asymmetric somatic hybridization between wheat (Triticum aestivum) and Avena sativa L. 总被引:1,自引:0,他引:1
XIANG Fengning XIA Guangmin CHEN Huimin 《中国科学:生命科学英文版》2003,46(3):243-252
Protoplasts from cell suspensions of young-embryo-derived calli, which were nonregenerable for long-term subculture and protoplasts from embryogenic calli with the regeneration capacity of 75% of the same wheat Jinan 177, were mixed as recipient. Protoplasts from embryogenic calli of Avena sativa (with the regeneration capacity of less than 10%) irradiated with UV at an intensity of 300 μW/cm2 for 30 s, 1 min, 2 min, 3 min, 5 min were used as the donor. Protoplasts of the recipient and the donor were fused by PEG method. Many calli and normal green plants were regenerated at high frequency, and were verified as somatic hybrids by chromosome counting, isozyme, 5S rDNA spacer sequence analysis and GISH (genomic in situ hybridization). Fusion combination between protoplasts either from the cell suspensions or from the calli and UV-treated Avena sativa protoplasts could not regenerate green plants. 相似文献
10.
V. M. Samoylov K. C. Sink 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(7):850-857
Highly asymmetric somatic hybrid plants were obtained by PEG/DMSO fusion of gamma-irradiated mesophyll protoplasts of the kanamycin-resistant (KmR+) interspecific hybrid Lycopersicon esculentum x L. pennellii (EP) with mesophyll protoplasts of Solanum melongena (eggplant, E). Elimination of the EP chromosomes was obtained by irradiating the donor genome with different doses of gamma rays (100, 250, 500, 750 and 1000 Gy). The selection of somatic hybrid calli was based on kanamycin resistance; EP and E protoplasts did not divide due to the irradiation treatment and sensitivity to kanamycin, respectively. KmR+ calli were recovered following all irradiation doses of donor EP protoplasts. The hybrid nature of the recovered calli was confirmed by PCR amplification of the NptII gene, RAPD patterns and Southern hybridizations using potato ribosomal DNA and pTHG2 probes. Ploidy levels of calli confirmed as hybrid were further analyzed by flow cytometry. Such analyses revealed that the vast majority of hybrid calli that did not regenerate shoots were 5–9n polyploids. The three asymmetric somatic hybrid plants obtained were regenerated only from callus with a ploidy level close to 4n, and such calli occurred only when the donor EP had been exposed to 100 Gy. The amount of DNA in somatic hybrid calli, from 100-Gy exposure, was found by dot blot hybridization with the species-specific probe, pTHG2, to be equivalent with 3.1–25.8% of the tomato genome. Thus, DNA contained in 3.8–13.2 average-size tomato chromosomes was present in these hybrid calli. The asymmetric somatic hybrid plants had the eggplant morphology and were regenerated from one hybrid callus that contained an amount of tomato DNA equivalent to 6.29 average-size tomato chromosomes. 相似文献
11.
N. B. Jelodar N. W. Blackhall T. P. V. Hartman D. S. Brar G. Khush M. R. Davey E. C. Cocking J. B. Power 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1999,99(3-4):570-577
Somatic hybrid plants were obtained following the electrofusion of rice (Oryza sativa L. cv ’Taipei 309’, 2n = 2x = 24) cell suspension–derived protoplasts with non-dividing leaf protoplasts of Porteresia coarctata (2n = 4x = 48), a saline-tolerant wild species. Fusion-treated protoplasts were plated on the surface of cellulose nitrate
filter membranes, overlaying Lolium multiflorum nurse cells. The nurse cells were embedded in KPR medium containing 0.5 mg l−1 2,4–dichlorophenoxyacetic acid and semi-solidified with SeaPlaque agarose. Putative somatic hybrid cell colonies were selected
on the basis of their growth, whereby faster growing colonies were transferred preferentially to MS-based medium with 2.0
mg l−1 kinetin, 0.5 mg l−1α-naphthaleneacetic acid, 30 g l−1 sucrose and 4.0 g l−1 SeaKem agarose to induce shoot regeneration. One hundred and nineteen regenerated plants were micropropagated clonally on
MS-based medium containing 2.0 mg l−1 6–benzylaminopurine, 50 g l−1 sucrose and 4.0 g l−1 SeaKem agarose, prior to DNA extraction of plant samples. Putative somatic hybrids were initially identified by RAPD analysis,
and 8 plant lines were selected for further investigation by flow cytometric ploidy determination and cytology. Plants of
one line had an allohexaploid chromosome complement (2n = 6x = 72) and, following examination of its vegetative clones by
GISH, were confirmed as somatic hybrids containing full chromosome complements of both O. sativa and P. coarctata.
Received: 27 July 1998 / Accepted: 19 December 1998 相似文献
12.
K. J. Puite J. G. Schaart 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(2-3):237-244
Summary The production of asymmetric somatic hybrid calli after fusion between gamma-irradiated protoplasts from transgenic Solanum brevidens and protoplasts from S. tuberosum are reported. Transgenic (kanamycin-resistant, GUS-positive) S. brevidens plants and hairy root clones were obtained after transformation with Agrobacterium tumefaciens LBA 1060 (pRi1855) (pBI121) and LBA 4404 (pRAL4404) (pBI121), and A. rhizogenes LBA 9402 (pRi1855) (pBI121), respectively. Leaf protoplasts isolated from the transgenic plants or root protoplasts from the hairy root clones were fused with S. tuberosum leaf protoplasts, and several calli were selected on kanamycin-containing medium. The relative nuclear DNA content of the hybrid calli was measured by flow cytometry (FCM), and the percentages of DNA of the S. brevidens and S. tuberosum genomes in the calli were determined by dot blot analysis using species-specific DNA probes. Chromosome-specific restriction fragment length polymorphism (RFLP) markers were used to investigate the elimination of specific S. brevidens chromosomes in the hybrids. The combined data on FCM, dot blot and RFLP analysis revealed that 18–62% of the S. brevidens DNA was eliminated in the hybrid calli and that the RFLP marker for chromosome 7 was absent in seven out of ten calli. The absence of RFLP markers for chromosomes 5 and 11 hardly ever occurred. In most of the hybrids the ploidy level of the S. tuberosum genome had increased considerably. 相似文献
13.
S. Krasnyanski T. M. Ball K. C. Sink 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(5):683-687
Twenty eight somatic hybrid plants were identified following protoplast fusions between peppermint (Mentha piperita L. cv Black Mitcham), producing high-quality oil, and spearmint (Mentha spicata L. cv Native Spearmint), likewise producing high-quality oil and also possessing resistance to verticillium wilt. Prior to
fusion, peppermint protoplasts were subjected to iodoacetic acid to inhibit cell division. Protoplasts of peppermint and spearmint
were fused using polyethylene glycol plus DMSO. Fusion products were cultured according to an efficient protoplast-to-plant-cycle
protocol developed for peppermint. Using this protocol, iodoacetic acid-treated peppermint protoplasts were not able to divide,
whereas untreated spearmint protoplasts had the ability to produce callus but not shoots. Therefore, selection of somatic
hybrid calli was based on the presumed capability of hybrid cells to form calli and shoots. Shoots in vitro were initially
identified as hybrids using RAPD profiles. Subsequently, observations on morphology, chromosome counts, and Southern-hybridization
patterns confirmed their hybrid status. The results of verticillium tests revealed that 18 somatic hybrids were more susceptible
than Native Spearmint, while hybrid II-14 had a level of susceptibility intermediate between that of the fusion parents. Oil-analysis
of hybrid plants indicated that they all have a GC-profile typical of spearmint oil.
Received: 8 February 1997 / Accepted: 9 April 1997 相似文献
14.
Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2011,106(3):391-399
Somatic embryogenesis (SE) was induced in female flower buds from mature Schisandra chinensis cultivar ‘Hongzhenzhu’. Somatic embryo structures were induced at a low frequency from unopened female flower buds and excised
unopened on Murashige and Skoog (MS) agar medium containing 4.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Friable embryogenic calli were induced from somatic embryo structures after three
to four subcultures on initiation medium. The frequencies of mature somatic embryo germination and plantlet conversion were
low, but increased in the presence of gibberellic acid (GA3). Some germinated somatic embryos could form friable embryogenic calli on medium without plant growth regulators (PGRs).
The germination and conversion frequencies of somatic embryos from embryogenic calli induced using PGR-free medium were higher
than for somatic embryos from embryogenic calli induced on medium containing 2,4-D. Most somatic embryos from 2,4-D-induced
embryogenic calli had trumpet-shaped embryos, and most somatic embryos from PGR-free medium–induced embryogenic calli had
two or three cotyledons. Histological observation indicated that two- and three-cotyledon embryos had defined shoot primordia,
but most of the trumpet-shaped embryos yielded plantlets that lacked or had poorly developed meristem tissue. Cytological
and random amplification of polymorphic DNA (RAPD) analyses indicated no evidence of genetic variation in the plantlets of
somatic embryo origin. 相似文献
15.
Xi-yan Yang Xian-long Zhang Shuang-xia Jin Li-li Fu Ling-gang Wang 《Plant Cell, Tissue and Organ Culture》2007,89(2-3):225-235
Asymmetric somatic hybrids were obtained between Gossypium hirsutum Coker 201 and wild cotton G. klozschianum Anderss. An investigation on the effect of ultraviolet (UV) irradiation on donor protoplasts was carried out, and the lethal
dose was determined to be 38.7 J cm−2. We firstly screened the putative hybrids by the color of the calli produced, followed by morphological, cytological, and
molecular analysis of putative hybrid plants. Most regenerated plants derived from fused protoplasts displayed a recipient-like
morphology, while some showed an intermediate phenotype between Coker 201 and G. klozschianum. Chromosome numbers in these somatic hybrids ranged from 54 to 74. The hybrids were verified by random amplified polymorphic
DNA (RAPD) and simple sequence repeat (SSR). Absence or co-existence of parents’ genome DNA fragments was identified through
molecular analysis. The heredity of cytoplasm was investigated by cleaved amplified polymorphic sequence (CAPS) analysis using
mitochondrial and chloroplast universal primer pairs. The results indicated that recombination and rearrangements might have
occurred in some regions of mitochondria (mt) and chloroplast (cp) DNA. To our knowledge, this is the first report about asymmetric
protoplast fusion in cotton, and the hybrids obtained would be useful for breeding programs. 相似文献
16.
J. Buiteveld Y. Suo M. M. Lookeren Campagne J. Creemers-Molenaar 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1998,96(6-7):765-775
Results are reported on the production and characterization of somatic hybrids between Allium ampeloprasum and A. cepa. Both symmetric and asymmetric protoplast fusions were carried out using a polyethylene-based mass fusion protocol. Asymmetric
fusions were performed using gamma ray-treated donor protoplasts of A. cepa and iodoacetamide-treated A. ampeloprasum protoplasts. However, the use of gamma irradiation to eliminate or inactivate the donor DNA of A. cepa proved to be detrimental to the development of fusion calli, and thus it was not possible to obtain hybrids from asymmetric
fusions. The symmetric fusions yielded a high number of hybrid calli and regenerated plants. The analysis of the nuclear DNA
composition using interspecific variation of rDNA revealed that most of the regenerated plants were hybrids. Flow cytometric
analysis of nuclear DNA showed that these hybrid plants contained a lower DNA content than the sum of the DNA amounts of the
parental species, suggesting that they were aneuploid. A shortage of chromosomes in the hybrids was confirmed by genomic in
situ hybridization. Chromosome counts in metaphase cells of six hybrids revealed that these plants lacked 2–7 leek chromosomes.
One hybrid showed also the loss of onion chromosomes. The hybrids had an intermediate phenotype in leaf morphology. The application
of these somatic hybrids in breeding is discussed.
Received: 7 April 1997 / Accepted: 10 September 1997 相似文献
17.
The mesophyll protoplasts were isolated from the Solanum tuberosum (S. tbr) clones of different ploidy level (4x Bzura cv., 2x H-8105, and 2x ZEL-1136) as well as from the wild species: S. bulbocastanum (S. blb, 2x) and two accessions of S. nigrum (S. ngr, 6x). Additionally, the protoplasts were isolated from the cell suspensions of Bzura cv. and H-8105 clone. The conditions
of protoplast isolation as well as the media for their culturing and regeneration, were selected and optimized for the studied
genotypes. For mesophyll protoplasts, the shooting calli were produced by all the cultured protoclones except that of S. bulbocastanum. The shoots excised from the protoplast-derived calli developed into whole plants in all the studied potato clones but only
in one accession of S. nigrum, i.e. S. ngr var. gigantea. As for suspension-cell-derived protoplasts, only H-8105 clone produced the regenerative type of calli, though normal shoots
could not be obtained. The regenerative capacity of the protoplasts isolated from leaves and cell suspensions is compared
and discussed.
We regret to report the death of M. Sc. Maria Borkowska after the completion of this work. 相似文献
18.
G. W. Bates 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1990,80(4):481-487
Summary Genetically asymmetric hybrids were recovered by fusion of Nicotiana tabacum protoplasts with irradiated protoplasts of kanamycin-resistant, nopalineproducing plants of N. repanda. Hybrid calli were selected by culture on media containing kanamycin and were regenerated. These plants were morphologically similar to N. tabacum but produced nopaline, indicating they retained genes from N. repanda. Esterase isozyme profiles also indicated that the plants are somatic hybrids, but are more similar to N. tabacum than N. repanda. Chromosome counts showed most of the hybrids had 55–62 chromosomes, which is consistent with extensive, although incomplete elimination of N. repanda chromosomes. The hybrids were largely male sterile, but about half of them set seed when crossed with N. tabacum. Chromosome numbers of the progeny and the pattern of inheritance of kanamycin resistance indicated the continued elimination of N. repanda genetic material in these backcrosses. The N. repanda parent used in these fusions gave a hypersensitive response to TMV, whereas the N. tabacum parent was TMV sensitive. When inoculated with TMV, plants from two hybrid clones gave a hypersensitive response. Plants from the other clones became systemically infected with the virus. 相似文献
19.
Protoplasts isolated from cotyledon-derived callus of Actinidia chinensisPlanch. var. chinensis (2N=2x=58) were fused with mesophyll protoplasts of Actiniadia kolomikta(Maxim. et Rupr.) Maxim (2N=2x=58) using a PEG method. Plantlets were regenerated from the fusion product clone 11. RAPD analyses, chromosome numbers of root tip cells and fluorescence peak position of leaf nuclei confirmed that clone 11 was an interspecific somatic hybrid (2N=4x=116) between A. chinensis and A. kolomikta. The chilling tolerance of the somatic hybrid was tested with in vitro leaves at low temperatures. Based on data of leaf thickness, electroconductivity, proline levels, malondialdehyde content and activity of superoxide dismutase, dendrogram cluster analysis suggested that the interspecific somatic hybrid was similar to A. kolomikta, and might have a higher capacity of cold resistance than A. chinensis. 相似文献
20.
M. A. O'Connell M. R. Hanson 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1985,70(1):1-12
Summary Selection and screening methods were devised which resulted in the identification of a number of somatic hybrid callus clones following fusion of Lycopersicon esculentum protoplasts and L. pennellii suspension culture protoplasts. Visual selection for callus morphology combined with a high fusion frequency and irradiation of one parental protoplast type (137Cs source, 1.5 Krads) resulted in selection of a callus clone population containing a high proportion of somatic hybrids. Analysis of a dimeric isozyme for the presence of a heterodimeric form was found to be satisfactory for distinguishing parental-type calli, somatic hybrid calli, and mixed calli derived from both types of unfused parental cells. No somatic hybrid calli produced shoots, although the sexual hybrid between L. esculentum and L. pennellii regenerated well under the culture conditions employed. This result suggests that the non-regenerable growth habit of the L. pennellii suspension culture was dominant in the somatic hybrid. The culture conditions described here are suitable for obtaining regenerated plants from L. esculentum mesophyll protoplasts. L. esculentum protoplast calli from fusion cultures gave rise to shoots with L. esculentum phenotype at higher frequency than calli from control unfused L. esculentum mesophyll protoplast cultures. The use of probes for species-specific organelle DNA fragments allowed identification of organelle DNA restriction fragments in digests of total DNA from small samples of individual callus clones. The callus clones analyzed either carried predominantly one parental plastid DNA type or mixtures of both types. Use of a mitochondrial DNA (mtDNA) probe which distinguishes two parental mtDNA fragments revealed that the L. pennellii-specific fragment was present in all clones examined, but the L. esculentum fragment was absent or in low proportion. 相似文献