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1.
Summary Primary callus of barley (Hordeum vulgare L.) derived from scutella (cv. Dissa) and anthers (cv. Igri) was used for protoplast isolation and plant regeneration. The protoplasts were embedded in agarose and cultured with nurse cells. The plating efficiency varied from 0.1% to 0.7%. Shoots regenerated from the developing callus. Plantlets were transferred to soil and cultivated in the greenhouse three to five months after protoplast isolation. All plants were normal in morphology, and most of them flowered and set seeds. 相似文献
2.
Wen-Ze Li Zi-Hong Song Bao-Tai Guo Li-Juan Xu 《In vitro cellular & developmental biology. Plant》2001,37(5):605-608
Summary The effects of DNA hypomethylating drugs (azacytidine and ethionine) on induction of microspore-derived calluses and embryos
were studied in barley (Hordeum vulgare L.) ev. Igri. The results were as follows: (1) Yield of calluses and embryos pretreated with the different concentrations
of azacytidine for 3 d was several-fold higher than that of the control. The highest yield of calluses and embryos in all
treatments appeared at a concentration of 3 mg l−1, which reached 11.03 per anther. It was 110-fold higher than the control. (2) There was a significant difference in yield
of calluses and embryos between the different days of pretreatment. The highest yield was obtained at a 3-d pretreatment.
If the period of pretreatment was shorter or longer than 3 d, yield of calluses and embryos was reduced sharply, and was similar
to that of the control. (3) The data obtained with ethionine pretreatment were very similar to those obtained with azacytidine.
(4) Tests on the different methods of pretreatment showed that yield of calluses and embryos pretreated with distilled H2O, mannitol, azacytidine, and ethionine was much higher than other pretreatments and the control, and reached 6.53–11.39 per
anther. The yield of calluses and embryos pretreated with DNA hypomethylating drugs was higher than with mannitol. However,
pretreatment with hypomethylation drugs supplemented with induction medium was not effective. 相似文献
3.
Summary A highly regenerable target tissue and a high-frequency DNA delivery system are required for the routine production of transgenic
barley. This project separately optimized tissue culture and particle bombardment parameters. Immature zygotic embryos (0.7
to 1.2 mm) were excised and culture on B5L solid medium. Klages and H930-36 cultivars regenerated significantly more green
plants than Sabarlis and Bruce. The regeneration pathway shifted from organogenesis to somatic embryogenesis when maltose
was used as the medium carbohydrate source instead of sucrose. More somatic embryos were induced on 5 mg/liter 2,4-dichlorophenoxyacetic
acid than 2 mg/liter. Gene delivery was optimized using anthocyanin regulatory genes as a transient marker. A 3-mm rupture
disc-to-macrocarrier gap distance, a 1-day prebombardment embryo culture period, and a maltose carbohydrate source were each
significantly better than other treatments. Double bombardments per plate, a 6-mm macrocarrier fly distance, and 650-psi rupture
discs each had the highest number of transiently expressing cells in individual experiments, although the results were not
statistically significant compared to the other treatments. Using the optimized parameters, over 200 cells routinely expressed
anthocyanin in a bombarded immature embryo. In tissue culture experiments, 350 to 400 green plants regenerated per 100 immature
embryos. The improvement of green plant regeneration and gene delivery forms a strong basis to develop a practical barley
transformation system. 相似文献
4.
R. Lührs H. Lörz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1987,75(1):16-25
Summary Immature embryos of 41 lines of barley were screened in vitro for callus induction and somatic embryogenesis on different media to establish totipotent cultures. The use of modified MS and CC media, both supplemented with 1 g/l casein hydrolysate, and the substitution of agarose for agar resulted in the highest frequencies of somatic embryo induction. Embryogenic callus was induced and plants regenerated from 23 of the lines tested. The auxins 2,4-D, dicamba, picloram and 2,4,5-T were suitable for embryogenic callus induction. High frequencies of somatic embryo germination occurred on CC medium supplemented with 1 mg/l IAA and 0.05 mg/l zeatin. A strong genotypic effect on the capacity and frequency of embryogenic callus formation was found. Cultivar Golden Promise always gave the best results. Experiments with field grown material in 3 consecutive years showed that environmental factors also strongly influenced the induction of somatic embryogenesis and plant regeneration.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid
- dicamba
3,6-dichloro-o-anisic acid
- picloram
4-amino-3,6,6-trichloropicolinic acid
- NAA
naphtaleneacetic acid
- IAA
indole-3-acetic acid
- ABA
abscisic acid
- BAP
6-benzyl amino purine
- 2iP
6-(3-methyl-2 butenyl 1-amino)purine
- GA3
gibberellic acid 相似文献
5.
P. R. Mark Shannon Anne E. Nicholson Jim M. Dunwell D. Roy Davies 《Plant Cell, Tissue and Organ Culture》1985,4(3):271-280
Barley anthers from cold pretreated spikes produced no or few calluses when plated with both loculi in contact with the medium (flat). When anthers were plated with only one loculus in contact with the medium (up), a high proportion of the anthers produced calluses. The top loculus of the up anthers was most productive. Flat anthers, when compared with up anthers, were not only slower to produce multicellular pollen grains (MCPs) and microcalluses, but also produced fewer of them and ceased production earlier. The MCPs and microcalluses in flat anthers grew more slowly and few developed beyond the 30 cell stage. These results establish the importance of anther orientation for barley anther culture. 相似文献
6.
Callus cultures were initiated from apical meristem explants of one to four-week-old aseptically-grown barley (Hordeum vulgare L. cv. Atlas 57) plants. Embryogenic callus and plants were produced in three separate experiments; the cultures have retained regenerative capacity for three years after initiation. Our results demonstrate that explants other than immature embryos are embryogenically competent in barley and that regeneration occurs by both somatic embryogenesis and organogenesis. 相似文献
7.
The radiation (32P) induced dwarf mutant of spring barley,Hordeum vulgare L., Mut. Dornburg 576, was genetically studied by crosses with the mother variety and characterized by seedling assays and investigations on development and yield formation. In comparison to the normal mother variety (Hordeum vulgare L., cv. Saale) mutant plants exhibit drastically reduced culm length, intensive tillering, a prolonged life cycle and a smaller biomass and grain yield formation. These characters are controlled by one gene in a pleiotropic way. The mutant responds with normal growth and development to the application of gibberellins. 相似文献
8.
X.-H. Wang P. A. Lazzeri H. Lörz 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1992,85(2-3):181-185
Summary Numerical and structural chromosome variation was analysed in dividing protoplasts isolated from suspension cells of barley. Five cell lines exhibited distribution patterns in chromosome number with different peaks and ranges. Embryogenic/morphogenic cell lines showed a peak at 2n = 14 (ca. 50%) after 6–7 months in culture, while older non-embryogenic cell lines had peaks at aneuploid or polyploid chromosome numbers. Culture duration had a clear effect on numerical and structural chromosome variation in embryogenic cell lines. With ageing of the cultures chromosome variation accumulated and the proportion of 2n = 14 cells decreased. The effect of protoplast isolation and culture on chromosome variation was examined; more cells with normal chromosome sets (12%) were maintained in protoplast-derived colonies than in source suspension cells (4%) of the same culture age.Abbreviations
DC
Dicentric
-
F
fragment
-
T
telocentric 相似文献
9.
The effects of water deficit on relative water content (RWC), on the activity superoxide dismutase (SOD) and peroxidase (POX)
from leaves of two drought-resistant barley strains (Hordeum vulgare L.) varieties (TOKAK-157/37 and 56000/MISC-233) and one sensitive (ERGINEL-90) were studied.
In 21 day old seedlings, drought stress was initiated by withholding water and lasted for 12 days. Activity of SOD increased
by the effect of drought treatments in the leaves of drought-resistant varieties TOKAK-157/37 and 56000/MISC-233 as compared
to sensitive variety ERGINEL-90. The drought treatment resulted in a 418 % and 59 % increase in SOD activity in resistant
varieties at the end of the 12th day of experimental period. However, an increase in activity of SOD was not accompanied by an increase in activity of POX
in drought-resistant TOKAK-157/37 and 56000/MISC-233 except on the 6th day of drought treatment in 56000/MISC-233. In drought-sensitive variety, ERGINEL-90, POX activity did not change throughout
drought period. 相似文献
10.
J. Schondelmaier R. Martin A. Jahoor A. Houben A. Graner H. -U. Koop R. G. Herrmann C. Jung 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1993,86(5):629-636
We have applied a refined microdissection procedure to create a plasmid library of the barley (Hordeum vulgare L.) chromosome arm 1HS. The technical improvements involved include synchronization of meristematic root tissue, a metaphase drop-spread technique, paraffin protection of the collection drop to avoid evaporation, and a motorized and programmable microscope stage. Thirteen readily-discernible telocentric chromosomes have been excised from metaphases of synchronized root-tip mitoses. After lysis in a collection drop (2 nl), the DNA was purified, restricted withRsaI, ligated into a vector containing universal sequencing primers, and amplified by the polymerase chain reaction. Finally, the amplified DNA was cloned into a standard plasmid vector. The size of the library was estimated to be approximately 44,000 recombinant plasmids, of which approximately 13% can be utilized for RFLP analysis. Tandem repetitive probes could be rapidly excluded from further analysis after colony hybridization with labelled total barley DNA. Analysis of 552 recombinant plasmids established that: (1) the insert sizes ranged between 70 and 1150 bp with a mean of 250 bp, (2) approximately 60% of the clones contained highly repetitive sequences, and (3) all single- or low-copy probes tested originate from chromosome 1HS. Four probes were genetically mapped, using an interspecificH. vulgare xH. spontaneum F2 population. One of these probes was found to be closely linked to theMla locus conferring mildew resistance. 相似文献
11.
R. B. Jørgensen B. Andersen 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1989,77(3):343-351
Summary The karyotype of 82 regenerated plants from callus cultures of interspecific hybrids between cultivated barley (Hordeum vulgare L.) and seven polyploid wild barley species was examined by C-banding or Feulgen staining. The karyotypic changes observed in 46 plants included aneuploidy, double haploidy, amphidiploidy, deletions, inversions, extra C-bands, and extra euchromatic segments. Apparently, chromosome 5, 6, and 7 of H. vulgare were more frequently exposed to elimination or structural change than the other chromosomes of this species. Irradiation of calli seemed to enhance the occurrence of karyotypic variants. 相似文献
12.
K. I. Gecheff 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1996,92(6):777-781
A total of 52 reciprocal translocations and 9 pericentric inversions were induced and identified in both standard and cytologically marked barley karyotypes using gamma-rays as the clastogenic agent. An analysis based upon Giemsa N-banding patterns and arm length measurements of the reconstructed chromosomes enabled a rather precise cytological localization of intra- and interchange breakpoints. This analysis was significantly facilitated and improved, especially for the identification of pericentric inversions, when the reconstructed karyotype T-1586 was used as starting material. The majority, if not all, of the aberration breakpoints proved to be localized in interband regions or in medial and terminal parts of the chromosomes, i.e., in regions which are deficient in constitutive heterochromatin. A great number of the structural mutations produced in this study contain specific cytological markers covering nearly all of the chromosomes of barley karyotype. This material might be of considerable interest in solving various problems of barley cytogenetics and chromosome engineering and especially in constructing a physical map of barley genome. 相似文献
13.
The influence of genotype and temperature pre-treatment on anther culture response in barley (Hordeum vulgare L.) 总被引:1,自引:0,他引:1
W. Powell 《Plant Cell, Tissue and Organ Culture》1988,12(3):291-297
The influence of temperature stress pre-treatment on anther culture response has been examined in eight commercially desirable barley cultivars. Spikes were pre-treated in darkness at 4°C for periods of 0, 7, 14, 21 and 28 days. Overall, the optimum pre-treatment period was 21 days, although there were large genotype by pre-treatment interactions. The most responsive cultivar was Igri, with a mean of 38% anthers responding, and relatively little effect of pre-treatment. The greatest effect of pre-treatment was in cv. Heriot, which had 3% response with no pre-treatment and 52% response from 14 days pre-treatment. 相似文献
14.
J. Zhang V. K. Tiwari T. J. Golds N. W. Blackhall E. C. Cocking B. J. Mulligan J. B. Power M. R. Davey 《Plant Cell, Tissue and Organ Culture》1995,41(2):125-138
Cat gene expression has been investigated following PEG-mediated plasmid uptake into barley protoplasts. The uptake conditions optimised for transient expression were employed for stable transformation. Transformed protoplast-derived calli of the cvs. Dissa and Igri, were selected on medium containing G418 at 40 g ml–1 or kanamycin sulphate at 250 g ml–1. Absolute transformation frequencies of 28.9×10–5 and 21.3×10–5 were recorded for Dissa with kanamycin sulphate and G418 selection, respectively. The frequency for Igri was 11.5×10–5 with G418 selection. Antibiotic resistant protoplast-derived colonies expressed NPTII activity; Southern hybridisation confirmed integration of the nptII gene into barley genomic DNA.Abbreviations ABA
abscisic acid
- AC-CAP
acetylated chloramphenicol
- BAP
6-benzylaminopurine
-
cat
chloramphenicol acetyltransferase gene
- CAT
chloramphenicol acetyltransferase activity
- CaMV
cauliflower mosaic virus
- CAP
chloramphenicol, 2,4-d-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- G418
Geneticin
-
gus
-glucuronidase gene
- HEPES
(N[2-hydroxyethyl] piperazine-N-[2-ethanesulphonic acid])
- IAA
indole acetic acid
- MES
2-N-morpholinoethane sulphonic acid
- NAA
-naphthaleneacetic acid
-
npt II
neomycin phosphotransferase gene
- NPTH
neomycin phosphotransferase activity
- PEG
polyethylene glycol
- SCV
settled cell volume 相似文献
15.
Anu Suoniemi Kesara Anamthawat-Jónsson Tiina Arna Alan H. Schulman 《Plant molecular biology》1996,30(6):1321-1329
The barley BARE-1 is a transcribed, copia-like retroelement with well-conserved functional domains, an active promoter, and a copy number of at least 3 × 104. We examined its chromosomal localization by in situ hybridization. The long terminal repeat (LTR) probe displayed a uniform hybridization pattern over the whole of all chromosomes, excepting paracentromeric regions, telomeres, and nucleolar organizer (NOR) regions. The integrase probe showed a similar pattern. The 5-untranslated leader (UTL) probe, expected to be the most rapidly evolving component, labeled chromosomes in a dispersed and non-uniform manner, concentrated in the distal regions, possibly indicating a targe site preference. 相似文献
16.
Cell-suspension cultures were initiated from embryogenic calli of various barley cultivars. Seven fast-growing suspension lines were obtained from four different cultivars (cvs. Dissa, Emir, Golden Promise and Igri). Two of these cell suspensions showed morphogenic capacity. From a cell suspension of cv. Dissa, albino plantlets were regenerated when aggregates were cultured on solid medium. Aggregates of cv. Igri usually stopped differentiation at the globular stage, but occasionally formed scutellum-like structures. Five suspension lines were used for protoplast isolation and culture. Dividing protoplasts were obtained from all lines, but with cv. Igri a few divisions only and no further development were observed. Protoplasts from the various lines differed in the time of first division (2–14 d), division frequency (0.09–70.9%) and efficiency of colony formation (0.09–7.3%). Protoplasts isolated from the morphogenic cell suspension of cv. Dissa developed compact calli which sporadically regenerated albino plantlets.Abbreviations CC, MS, N6, SH, Kao8p
culture media; see Material and methods
- cv
chltivar
- dicamba
3,6-dichloro-o-anisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
17.
A. K. Gaponenko T. F. Petrova A. R. Iskakov A. A. Sozinov 《TAG. Theoretical and applied genetics. Theoretische und angewandte Genetik》1988,75(6):905-911
Summary Cytogenetic analysis of immature embryoderived calli and regenerated plants of barley has demonstrated high heterogeneity of callus cultures and significant differences in cytogenetic processes between different callus lines. Regenerated plants usually have a normal chromosome complement (2n=14). Tetraploid plaints occur with a frequency of 1%. No chromosome aberrations have been detected by Feulgen staining. The phenomenon of chromosome stickiness recorded from the 2nd day of culture was discovered in a majority of callus lines as well as the phenomena of chromatin hypercondensation and chromosome supercoiling. A possible contribution of cytogenetic and molecular processes to somaclonal variation is discussed. 相似文献
18.
Ravindra N. Chibbar Kutty K. Kartha Raju S. S. Datla Nick Leung Karen Caswell Clifford S. Mallard Lee Steinhauer 《Plant cell reports》1993,12(9):506-509
The cauliflower mosaic virus 35S (35S) and the enhanced 35S (E35S) promoters fused with maize alcohol dehydrogenase (Adh1) intron1 or maize shrunken locus (sh1) intronl along with maize Adh1 and rice actin (Act1) promoters fused to their respective first introns were tested for transient expression of the E.coli -glucuronidase (gus) reporter gene in cultured barley (Hordeum vulgare L) cells. The plasmids, carrying the respective promoterintron combinations to drive the gus fused to nopaline synthase (nos) terminator, were introduced into cultured barley cells using a particle gun. The rice Act1 promoter with its first intron gave the highest expression of all promoter intron combinations studied. This was followed by the E35S promoter and no significant differences were observed between the other two promoters tested. The rice actin promoter is now being used to drive selectable marker genes to obtain stably transformed cereal cells.NRCC No. 36482 相似文献
19.
Tocotrienols are lipophilic antioxidants belonging to the tocochromanols, better known as vitamin E. Although present in cereal grains in high quantities not much is known about their function in plants. In a detailed study the temporal and spatial accumulation of tocotrienols and tocopherols during grain development in two barley cultivars was analyzed. Tocochromanols and lipids accumulated in parallel until 80% of the final dry weight of the kernels was reached. Later on the tocochromanol content did not change while the lipid content decreased. Generally, only about 13% of the tocochromanols were found in the germ fraction, whereas the pericarp fraction contained about 50% and the endosperm fraction about 37% of the tocochromanols. Altogether, about 85% of the tocochromanols were tocotrienols in both cultivars. In case of the tocopherols about 80% were found in the germ fraction and the remaining 20% in the pericarp fraction. Tocotrienols were almost equally present in the pericarp and the endosperm fraction. Individual forms of tocopherols and tocotrienols accumulated with different kinetics during barley grain development. The differences in distribution and accumulation indicate different functions of the individual tocochromanols during grain development. 相似文献
20.
Summary Variation in the nitrogen content of seed of six barley cultivars was brought about by growing parent plants at four nitrogen levels. Shoot dry weight of plants grown for 23 days from these seeds was generally enhanced by an increase in seed nitrogen content. The most responsive cultivar was a primitive type of barley from Ethiopia. Cultivars with a longer breeding history were less responsive. Risø 1508 apparently had physiological and biochemical limitations in responding to extra seed nitrogen. In the barley cultivars studied extra seed nitrogen seems to supplement, rather than substitute for, nitrogen fertilizer in the seed bed. 相似文献