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1.
The effect of streptomycin on morphogenic explants of Lycopersicon peruvianum Mill. was examined microscopically at both the light and ultrastructural level. Early stages in shoot regeneration from leaf explants were distinguished as meristematic tissue at both levels. Small starch grains were observed in the plastids in this tissue but not in plastids in regenerated shoots. In the presence of streptomycin, adventitious shoot regeneration from sensitive leaf strips was inhibited. Large layered bodies were observed within the plastids of sensitive leaf tissue, suggesting the disruption of thylakoid membrane formation. Streptomycin resistant L. peruvianum lines, as well as a chlorophyll-deficient line, were also examined microscopically. The chloroplasts of newly regenerated streptomycin resistant shoots contained well developed internal membranes and conspicuous starch grains. Cells containing a mixture of resistant and sensitive plastids were not observed. The plastids in chlorophyll-deficient tissue completely lacked thylakoid membranes, although small vesicles and intraplastid bodies were seen within the stroma.Abbreviations NMU
N-methyl-N-nitrosourea 相似文献
2.
Susanne Knapp George Coupland Helmut Uhrig Peter Starlinger Francesco Salamini 《Molecular & general genetics : MGG》1988,213(2-3):285-290
Summary The maize transposable element Ac has been introduced into potato via the T-DNA (transferred DNA) of Agrobacterium tumefaciens. Ac was inserted within the untranslated leader region of a neomycin phosphotransferase II (NPT-II) gene such that excision restored NPT-II activity. Two approaches to monitor Ac excision were used. (i) Using an Agrobacterium strain harbouring plasmid pGV3850::pKU3, leaf discs were selected on kanamycin (Km) after exposure to Agrobacterium. (ii) Using a strain containing plasmid pGV3850HPT::pKU3, the leaf discs were selected on hygromycin (Hm) and the resulting shoots were checked for NPT-II expression. Thirteen kanamycin resistant shoots transformed with pGV3850::pKU3 were isolated, suggesting that Ac had excised from the NPT-II gene. Out of 43 hygromycin resistant shoots transformed with pGV3850HPT::pKU3, 22 expressed the NPT-II gene, indicating that Ac had undergone excision in approximately 50% of the hygromycin resistant shoots. Southern analysis revealed that all kanamycin resistant plants contained the DNA restriction fragments expected when Ac excises from the NPT-II gene. The presence of Ac at new locations within the genomic DNA of several transformants was also detected. 相似文献
3.
A system has been developed for rapid selection of streptomycin resistant mutants, as adventitious shoots arising from explants of several Solanaceous species. Efficient mutagenesis was achieved by incubating shoot culture-derived leaf strips with 1 or 5 mM nitroso-methylurea, for 90 or 120 min. In Nicotiana tabacum and Lycopersicon peruvianum these treatments resulted in white or variegated adventitious shoots from up to 3.5% of explants placed on medium promoting shoot regeneration. Chlorophyll deficiencies were only observed very rarely in Solanum nigrum. Streptomycin resistant shoots were obtained from leaf explants placed on medium containing 500 mg l-1 streptomycin sulphate, under which conditions explants are bleached and adventitious shoot development suppressed. Green adventitious s shoots appeared at a frequency dependent both on the mutagenic treatment and on the species. The best response was with S. nigrum where >70% of the explants produced streptomycin resistant shoots, most of which retained their resistance on subsequent testing. Maternal inheritance of streptomycin resistance has been confirmed for several N. tabacum and S. nigrum mutants, and there is also evidence for paternal transmission in the latter species. The procedure has been successfully extended to other species, including N. sylvestris and N. plumbaginifolia, and also to obtain spectinomycin resistant mutants.Communicated by R. Hagemann 相似文献
4.
Streptomycin resistant shoots were regenerated from mutagenised (ethylemethane sulphonate treated) cotyledon explants ofCapsicum annuum Cv G4. Streptomycin resistant shoots developed from green (unbleached) sectors of the cotyledons. Reciprocal crosses betweeen streptomycin resistant and sensitive plants have shown a non-mendelian transmission. Such mutant plants should be useful in designing biochemical selection schemes to recover somatic hybrids and cybrids ofC. annuum.ABBREVIATIONS BAP
6-Benzylaminopurine
- IAA
Indoleacetic acid
- EMS
Ethyl methane sulphonate
- NMU
Nitrosomethyl urea 相似文献
5.
A high frequency of plastid-encoded antibiotic-resistant variants of Capsicum annuum were isolated on selective media following treatment by ethyl methanesulphonate (EMS) and nitrosomethylurea (NMU). Seeds
and explants were mutagenized with 0.1% EMS and 5 mm NMU separately. Non-mutagenized cotyledons (controls), mutagenized cotyledons from EMS-treated seedlings and NMU-treated
cotyledons were placed on regeneration medium supplemented with the antibiotics streptomycin or lincomycin. Resistant shoots
appeared at a high frequency in mutagenized cotyledons, whereas in controls morphogenesis was suppressed, accompanied by bleaching.
The stability of streptomycin and lincomycin resistance was confirmed by leaf assay. NMU-treated cotyledons gave a higher
frequency of variants than cotyledons from EMS-treated seedlings. The mutagenic effect of EMS was more pronounced using whole
seeds rather than cotyledons; in contrast, NMU was more effective in inducing variations in cotyledons than in seeds.
Received: 1 October 1996 / Revision received: 12 May 1997 / Accepted: 3 June 1997 相似文献
6.
D.J. Crook J. Cross A.N.E. Birch R.M. Brennan A.J. Mordue 《Entomologia Experimentalis et Applicata》2001,101(2):183-190
Monitoring of an unsprayed infested fieldsite using watertraps in S.E. Kent revealed four generations of Dasineura tetensi (Rubs) (Diptera: Cecidomyiidae) occurring between April and September 1996. Ribes nigrum L. cultivars 'Baldwin' (susceptible), 'Ben Alder' (susceptible) and 'Ben Connan' (resistant) were sampled for eggs in the field and assessed for midge damage throughout the season. Oviposition was indiscriminate, but plant damage varied significantly between cultivars. In laboratory choice experiments, mated female midges showed no preference between susceptible shoots of 'Ben Alder' and resistant shoots of 'Ben Connan' for oviposition. Olfactory responses of D. tetensi to leaf volatiles of 'Ben Alder' and 'Ben Connan' were also tested in a 4-way olfactometer. Mated females did not discriminate between volatiles of susceptible and resistant host plants. Larvae reared on cv. 'Ben Connan' shoots were significantly smaller than those reared on shoots of cv. 'Ben Alder'. Larval antibiosis and not female antixenosis appears to be the main mechanism for resistance to D. tetensi in 'Ben Connan'. 相似文献
7.
植物染色体的倍性维持和变化受环境因素影响,组培再生过程由于培养条件等因素往往导致染色体的结构和倍性变化。为探索组培条件下山茶种质的倍性变化,该研究利用山茶种质的愈伤组织诱导体系,通过流式细胞仪分析倍性变化情况,并结合秋水仙素处理对组培再生条件下倍性的稳定性和变化进行分析。结果表明:(1)10个山茶种质中6个为二倍体,2个为四倍体,1个为六倍体和1个为十倍体,在组培诱导愈伤及再生过程中不同倍性的种质材料能够保持稳定的倍性。(2)获得了秋水仙素处理的最适诱导条件,即培养基配方为秋水仙素浓度20 mg·L-1,愈伤增殖培养10 d。(3)对56个独立组织样品(含愈伤和芽)开展了倍性检测发现,有38个组织样品的倍性在1.5~2.5倍之间,11个组织样品产生了低于1.5倍性的特异现象。该研究结果进一步探索了不同山茶种质之间的倍性关系,为山茶属植物的倍性调控和多倍体诱导提供了理论基础。 相似文献
8.
The species Solanum surattense Burm.f. has importance in ayurvedic medicine and also as vegetable. Streptomycin-resistant plantlets were induced showing chloroplast encoded mutants in S. surattense from mutagenised (ethyl methane sulphonate and gamma-rays) cotyledon explants. Chloroplast encoded – streptomycin resistant – shoots were developed from green (unbleached) sectors of the cotyledons. The streptomycin-resistant plants were similar to parental plants in morphology and ploidy level (2n=2x=24). Reciprocal crosses between streptomycin-resistant and the original streptomycin sensitive plants have shown the non-Mendelian transmission under the control of chloroplast – DNA. These antibiotic resistant plants are useful in designing biochemical selection schemes aimed at somatic hybrid/cybrid recovery in S. surattense. 相似文献
9.
Summary Leaf discs of C. intybus were inoculated with an Agrobacterium tumefaciens strain harboring a neomycin phosphotransferase (neo) gene for kanamycin resistance and a mutant acetolactate synthase gene (csr1-1) from Arabidopsis thaliana conferring resistance to sulfonylurea herbicides. A regeneration medium was optimized which permitted an efficient shoot regeneration from leaf discs. Transgenic shoots were selected on rooting medium containing 100 mg/l kanamycin sulfate. Integration of the csr1-1 gene into genomic DNA of kanamycin resistant chicory plants was confirmed by Southern blot hybridizations. Analysis of the selfed progenies (S1 and S2) of two independent transformed clones showed that kanamycin and chlorsulfuron resistances were inherited as dominant Mendelian traits. The method described here for producing transformed plants will allow new opportunities for chicory breeding. 相似文献
10.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Pistacia vera L. cv. Siirt. The in vitro procedure involved four steps that included (1) induction of shoot initials from the regenerated mature leaf tissue, (2)
regeneration and elongation of shoots from the shoot initials, (3) rooting of the shoots, and (4) acclimatization of the plantlets.
The induction of shoot initials was achieved on an agarified Murashige and Skoog (MS) medium with Gamborg vitamins supplemented
in different concentrations of benzylaminopurine (BA) and indole-3-acetic acid (IAA). The best medium for shoot induction
was a MS medium with 1 mgl−1 IAA and 2 mgl−1 BA. Numerous shoot primordia developed within 2–3 wk on the leaf margin and the midrib region, without any callus phase.
In the second step, the shoot clumps were separated from the leaf explants and transferred to a MS medium supplemented with
1 mgl−1 BA, resulting in a differentiation of the shoot initials into well-developed shoots. The elongated shoots (>3 cm long) were
rooted on a full-strength MS basal medium supplemented with 2 mgl−1 of indole-3-butyric acid in the third stage. Finally, the rooted plants were transferred to soil with an 80% success rate.
This protocol was utilized for the in vitro clonal propagation of this important recalcitrant plant species. 相似文献
11.
Highly regenerative callus was isolated from the base of adventitious shoots on cotyledon explants of Dianthus hybrida Telstar Scarlet cultured on MS medium supplemented with 1 mg l−1 TDZ and 0.1 mg l−1 NAA. Flow cytometric analysis showed that cotyledon tissue is a mixture of diploid and tetraploid cells. Whereas the regenerative callus consisted of cells showing various ploidy levels of 2C to 16C, their regeneration ability was maintained as long as they were sub-cultured onto fresh media. More than 93% of regenerated shoots from the calluses were diploid. Only a few shoots were revealed as tetraploids and octoploids, suggesting that diploid cells had higher regeneration ability. 相似文献
12.
Regeneration of transgenic tamarillo plants 总被引:2,自引:0,他引:2
Media were developed to regenerate shoots from leaf pieces of tamarillo (Cyphomandra betacea (Cav.) Sendtner). Shoots were derived via organogenesis and could be easily rooted and transferred to the growth chamber. Transgenic tamarillo plants were produced using the binary vector pKIWI110 in the avirulent Agrobacterium strain LBA4404. All transgenic plants were kanamycin resistant and some plants expressed the D-glucuronidase (gusA) reporter gene and were chlorsulfuron resistant. Molecular evidence for transformation was obtained using PCR (polymerase chain reaction) and Southern hybridization. Inheritance of the transgenic phenotypes was demonstrated in seedling progeny. 相似文献
13.
Metinee Srivatanakul Sung Hun Park Maria G. Salas Roberta Hawkins Smith 《Journal of plant physiology》2001,158(2)
Kenaf(Hibiscus cannabinus)is a fast growing annual with tremendous potential as a source of fiber for ropes, textiles and paper. Kenaf is an environmentally friendly crop; however, commercial production of kenaf is hindered by weed competition at the seedling stage. Herbicide resistant kenaf cultivars would reduce seedling weed competion and make growing kenaf more profitable. Factors that are important in establishing a transformation system for kenaf were examined. The influence of Agrobacterium strain, temperature, host tissue wounding, acetosyringone, virG/virE genes and host cell division on T-DNA expression in the kenaf shoot apex were investigated. Three Agrobacterium strains were tested, and A. tumefaciens LBA4404 significantly (α=0.05) yielded a high number of shoots surviving on selection medium; no shoots survived with EHA101S or Z707S. There was no significant difference (α=0.05) in transient T-DNA expression between 28 °C and 25 °C; however, shoots did not survive 16 °C or 19 °C co-cultivation temperatures. Shoot apex survival was increased significantly (α=0.05) when virulence genes and a cytokinin, TDZ, were combined. Sonicated shoots showed an increase in transient expression and shoot survival. Optimal conditions for shoot apex T-DNA transfer and expression were sonication for 5 s, co-cultivation with LBA4404 containingvirG/virEat room temperature, and 200 μmol/L acetosyringone. 相似文献
14.
Photosynthesis,Transpiration, and Water Use Efficiency of Vegetative and Reproductive Shoots of Grassland Species from North-Eastern China 总被引:2,自引:0,他引:2
The differences in net photosynthetic rate (P
N), transpiration rate (E), and water use efficiency (WUE) between the vegetative and reproductive shoots of three native grass species from the grassland of northeastern China [grey-green and yellow green populations of Leymus chinensis (Trin.) Tzvel., Puccinellia tenuiflora (Griseb) Scrib & Merr, Puccinellia chinampoensis Ohwi] were compared. The two type shoots experienced similar habitats, but differed in leaf life-span and leaf area. The leaf P
N and WUE for the vegetative shoots were significantly higher than those for the reproductive shoots in the grasses, while their E were remarked lower in the dry season. Relative lower leaf P
N and WUE for the reproductive shoots of grassland grasses may explain the facts of lower seed production and the subordinate role of seed in the grassland renewal in north-eastern China. 相似文献
15.
P. F. McCabe L. J. Dunbar A. Guri K. C. Sink 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(2-3):377-382
Summary Asymmetric somatic hybrid plants were recovered after fusing irradiated mesophyll protoplasts of donor Lycopersicon esculentum × L. pennellii (EP) interspecific hybrid with callus-derived protoplasts of recipient Solanum lycopersicoides. EP plant A54 had been previously transformed by an agrobacterium vector, and the T-DNA insert mapped to the L. esculentum chromosome 12. The T-DNA insert conferred kanamycin resistance to EP that was subsequently used to select cell fusion products and recover asymmetric hybrid plants that retained tagged chromosome 12. Doses of 50- and 100-Gy irradiation promoted the elimination of only a few donor chromosomes. At 200 Gy, the regenerated plants had ploidy levels higher than tetraploid. However, the T-DNA tagged chromosome 12 was always retained in the asymmetric hybrid plants tested. Likewise, all plants from the 100-Gy series, with the exception of number 160, were mixoploid in the root-tip cells. Such mixoploid asymmetric somatic hybrids could be stabilized by inducing adventitious shoots on leaf strips cultured on shoot regeneration medium containing kanamycin. The asymmetric hybrid plants did not produce viable seed when self-pollinated or backcrossed to tomato or S. lycopersicoides.
Present address: Department of Biology, University College of London, Gower Street, London, UK 相似文献
16.
Hiroki Takagi Shintaro Sugawara Tomoka Saito Haruka Tasaki Lu Yuanxue Guan Kaiyun Dong-Sheng Han Toshinari Godo Masaru Nakano 《Plant biotechnology reports》2011,5(2):187-195
The gesneriaceous perennial plant Titanotrichum oldhamii has beautiful foliage and attractive bright yellow flowers. However, breeding of T. oldhamii by conventional sexual hybridization may be difficult because sexual reproduction of this species is very rare. In the present
study, plant regeneration systems via both direct and indirect formation of adventitious shoots from leaf explants were established
as the first step toward breeding T. oldhamii by using biotechnological techniques. Adventitious shoots were formed efficiently on medium containing 0.1 mg l−1 benzyladenine. Histological observation showed that shoot formation on this medium occurred directly from leaf epidermal
cells without callus formation. On the other hand, leaf explants formed calluses on medium containing 0.1 mg l−1 2,4-dichlorophenoxyacetic acid. The calluses could be maintained by monthly subculturing to fresh medium of the same composition.
When the calluses were transferred to plant growth regulator-free medium, they formed adventitious shoots. Directly and indirectly
formed shoots rooted well on medium containing 0.1 mg l−1 indole-3-butyric acid. Plantlets thus obtained were successfully acclimatized and grew vigorously in the greenhouse. Flow
cytometry analysis indicated that no variation in the ploidy level was observed in plants regenerated via direct shoot formation,
whereas chromosome doubling occurred in several plants regenerated via indirect shoot formation. Regenerated plants with the
same ploidy level as the mother plants showed almost the same phenotype as the mother plants, whereas chromosome-doubled plants
showed apparent morphological alterations: they had small and crispate flowers, and round and deep green leaves. 相似文献
17.
Public concern and metabolic drain were the main driving forces for the development of a selectable marker-free transformation
system. We demonstrated here the production of transgenic tobacco plants using a non-selection approach by Agrobacterium tumefaciens-mediated transformation. A. tumefaciens-infected leaf explants were allowed to produce shoots on a shoot induction medium (SIM) containing no selective compounds.
Up to 35.1% of the A. tumefaciens-infected leaf explants produced histochemically GUS+ shoots, 3.1% of regenerated shoots were GUS+, and 72% of the GUS+ shoots were stably transformed by producing GUS+ T1 seedlings. When polymerase chain reaction (PCR) was used to screen the regenerated shoots, 4% of the shoots were found
to be PCR+ for the transgene and 65% of the PCR+ shoots were stable transformants. Also, generation of PCR+ escapes decreased linearly as the number of subculture increased from one to three on SIM containing the antibiotic that
kills the Agrobacterium. Twenty-five to 75% of the transformants were able to transmit transgene activity to the T1 generation in a Mendelian 3:1
ratio, and a transformation efficiency of 2.2–2.8% was achieved for the most effective binary vector. These results indicated
that majority of the GUS+ or PCR+ shoots recovered under no selection were stable transformants, and only one-third of them were chimeric or escapes. Transgenes
in these transgenic plants were able to transmit the transgene into progeny in a similar fashion as those recovered under
selection. 相似文献
18.
Tomoko Igawa Yoichiro Hoshino Masahiro Mii 《In vitro cellular & developmental biology. Plant》2002,38(2):157-162
Summary A plant regeneration system from cell suspension cultures was established in an important ornamental crop, Limonium sinuatum Mill. cv. ‘Early Rose’. Friable callus was initially induced from leaf segments of in vitro-cultured seedlings on 0.25% gellan gum-solidified half-strength Murashige and Skoog [1/2MS] medium containing 1.0 mg l−1 (4.14 μM) picloram. These calluses were maintained as cell suspension cultures, which showed high proliferation ability with about
80 times increase in fresh weight during the 2-wk interval of subculture. Shoot regeneration from these cell cultures was
achieved by cytokinins, especially zeatin, which was the most effective in producing normal shoots with reduced hyperhydration
when used in combination with 0.5% gellan gum. Shoot regeneration ability was different among the cell lines originated from
each different seedling. Shoot formation was observed at different frequencies on four of five cell lines whereas one cell
line showed no shoot differentiation. Regenerated shoots detached from callus readily rooted 1 mo. after the transfer onto
0.5% gellan gum-solidified 1/2MS medium lacking plant growth regulators. The plantets were successfully transferred to the
greenhouse after acclimatization. No ploidy changes were observed in the callus induced or in the regenerated plantlets. The
regenerated plantlets that were transferred to the greenhouse after acclimatization grew normally and did not any morphological
signs of somaclonal variation. 相似文献
19.
Somatic chromosome number doubling of selected potato genotypes using callus culture or the colchicine treatment of shoot nodes in vitro 总被引:1,自引:0,他引:1
Experiments were carried out to double the somatic cell chromosome numbers of a monoploid and dihaploid of Solanum tuberosum and a genotype of S. circaeifolium subsp. quimense. Colchicine was used in vitro on shoot nodes from which the axillary meristems had been removed. Plants with doubled chromosome numbers were obtained from shoots grown from the tertiary, sub-axillary meristems of all three genotypes. The callus culture of stem and leaf explants was found to produce more shoots with doubled chromosome numbers than the colchicine treatment in the case of the dihaploid and quimense genotypes but no shoots were obtained from callus culture of the monoploid. Fifty-two % of the shoots from the dihaploid and 63% from the quimense clone were ploidy doubled in the case of the best callus culture system. Using a sub-lethal dose of colchicine, the dihaploid yielded 37% ploidy-doubled shoots whereas all the shoots produced from the monoploid were doubled and the quimense clone produced 27% doubled plants. Callus culture was highly dependent upon the type of growth medium and other, unknown, factors. The colchicine treatment, although yielding fewer products, was more reliable for achieving ploidy doubling in selected clones if the number of plants produced is not important. 相似文献
20.
Summary An efficient protocol has been developed for the regeneration of plantlets from leaf explants of witloof chicory (Cichorium intybus L.). Regeneration via callus was obtained on modified Murashige and Skoog semisolid medium (MS) containing 2.0 μM indole-3-acetic acid +5.0 μM 6-furfurylaminopurine (kinetin), and 1000 mgl−1 casein hydrolyzate. At least five or more shoots regenerated from each callus. The shoots were rooted on MS +0.2 μM indole-3-butyric acid. The plantlets thus obtained were successfully established in soil after bardening. Esculin accumulation
was recorded in plant tissues at different stages of differentiation in in vitro cultures and compared with in vivo-grown, plants. The esculin accumulation was higher in in vitro plants. 相似文献