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1.
Plant regeneration from somatic embryogenic suspension cultures of cotton (Gossypium hirsutum L.) 总被引:1,自引:0,他引:1
John J. Finer 《Plant cell reports》1988,7(6):399-402
Maintainable, highly embryogenic suspension cultures of cotton (Gossypium hirsutum L. cv. Coker 310) have been obtained. Callus cultures were initiated from cotyledonary tissues from aseptically-germinated seedlings. To establish the suspension cultures, callus tissue was placed in a liquid medium containing either 0.5 mg/l picloram or 0.1 mg/l 2,4-dichlorophenoxyacetic acid. For proliferation of the embryogenic suspension, 5 mg/l of 2,4-dichlorophenoxyacetic acid was used. Embryo development took place when the embryogenic tissue was transferred to an auxin-free liquid medium containing 15 mM glutamine. Early embryo development was fairly synchronous and large numbers of somatic embryos were produced. Regenerated plants were fertile and smaller than seed-derived plants.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- IAA
indole-3-acetic acid 相似文献
2.
High-frequency stable transformation of cotton (Gossypium hirsutum L.) by particle bombardment of embryogenic cell suspension cultures 总被引:6,自引:0,他引:6
K. Rajasekaran R. L. Hudspeth J. W. Cary D. M. Anderson T. E. Cleveland 《Plant cell reports》2000,19(6):539-545
Stable transformation of cotton (Gossypium hirsutum L.) at a high frequency has been obtained by particle bombardment of embryogenic cell suspension cultures. Transient and
stable expression of the β-glucuronidase (GUS) gene was monitored in cell suspension cultures. Transient expression, measured 48 h after bombardment,
was abundant, and stable expression was observed in over 4% of the transiently expressing cells. The high efficiency of stable
expression is due to the multiple bombardment of rapidly dividing cell suspension cultures and the selection for transformed
cells by gradually increasing the concentrations of the antibiotic Geneticin (G418). Southern analysis indicated a minimum
transgene copy number of one to four in randomly selected plants. Fertile plants were obtained from transformed cell cultures
less than 3 months old. However, transgenic and control plants from cell cultures older than 6 months produced plants with
abnormal morphology and a high degree of sterility.
Received: 20 January 1999 / Revision received: 1 October 1999 / Accepted: 11 October 1999 相似文献
3.
Tanoh Hilaire Kouakou Pierre Waffo Téguo Josep Valls Yatty Justin Kouadio Alain Decendit Jean-Michel Mérillon 《Plant Cell, Tissue and Organ Culture》2006,86(3):405-409
For the first time, trans-resveratrol, a stilbene, has been identified in cotton cell suspensions. Cell suspensions of Coker 312, a cultivar which produces embryogenic structures, acccumulate trans-resveratrol contrary to those of cultivar R405-2000, which do not. This stilbene may be a good phenolic marker for induction of somatic embryogenesis in cotton. 相似文献
4.
A heterotrophic cotton (Gossypium hirsutum L. cv. Stoneville 825) cell suspension culture was adapted to grow photoautotrophically. After two years in continuous photoautotrophic culture at 5% CO2 (balance air), the maximum growth rate of the photoautotrophic cell line was a 400% fresh weight increase in eight days. The Chl concentration was approximately 500 g per g fresh weight.Elevated CO2 (1%–5%) was required for culture growth, while the ambient air of the culture room (600 to 700 ul CO2 1–1) or darkness were lethal. The cell line had no net photosynthesis at 350 ul 1–1 CO2, 2% O2, and dark respiration ranged from 29 to 44 mol CO2 mg–1 Chl h–1. Photosynthesis was inhibited by O2. The approximate 1:1 ratio of ribulose 1,5-bisphosphate carboxylase (RuBPcase) to phosphoenolpyruvate carboxylase (PEPcase) (normally about 6:1 in mature leaves of C3 plants) was due to low RuBPcase activity relative to that of C3 leaves, not to high PEPcase activity. The PEPcase activity per unit Chl in the cell line was identical to that of spinach leaves, while the RuBPcase activity was only 15% of the spinach leaf RuBPcase activity. RuBPcase activity in the photoautotrophic cells was not limited by a lack of activation in vivo, since the enzyme in a rapidly prepared cell extract was 73% activated. No evidence of enzyme inactivation by secondary compounds in the cells was found as can be found with cotton leaves. Low RuBPcase activity and high respiration rates are most likely important factors in the low photosynthetic efficiency of the cells at ambient CO2.Abbreviations Chl
chlorophyll
- COT
heterotrophic cotton cell line
- COT-P
photoautotrophic cotton cell line
- DCMU
3-(3,4-dichlorophenyl)-1,1-dimethylurea
- HEPES
N-2-hydroxyethylpiperazine-N-2-ethanesulfonic acid
- Rubisco
ribulose 1,5-bisphosphate carboxylase/oxygenase
- RuBP
ribulose 1,5-bisphosphate
- RuBPcase
RuBP carboxylase
- PEP
phosphoenolpyruvate
- PEPcase
phosphoenolpyruvate carboxylase
- MX
Murashige and Skoog medium with 0.4 mg 1–1 2,4-D
- KT
photomixotrophic medium with 1% sucrose
- KTo
KT medium with no carbohydrate
- KTPo
KTo medium supplemented with 0.3 M Picloram
- CER
CO2 exchange rate
- PCER
CO2 exchange rate in the light 相似文献
5.
在自然衰老和诱导条件下棉花悬浮细胞程序性死亡的发生 总被引:2,自引:0,他引:2
Cotton suspension cells grew well in the MS medium supplemented with 0.1 mg/L 2,4 D and 0.1 mg/L KT. Senescence occurred when the cells were unsubcultured. The cells began to lose their viabilities on the 17th day, and on the 21th day oligonucleosomal sized DNA fragments ( DNA ladder) could be detected. Oligonucleosomal sized DNA fragments ( DNA ladder) was the hallmark of the programmed cell death. Programmed cell death of cotton suspension cells could be induced respectively by some stress factors which included heatshock (42+/-3 degrees C for 8 hours), 10 micromol/L camptothecin, 20 micromol/L fumonisin B1 and 50 mmol/L cycloheximide. The cotton suspension cells which grew in the MS medium supplemented with 0.1 mg/L 2,4 D and 0.1 mg/L KT differred physiologically from the cells in the MS medium supplemented with 0.1 mg/L IBA and 0.1 mg/L KT, and they responded differentially to the heatshock, 10 micromol/L camptothecin and 20 micromol/L fumonisin B1, while the same to 50 mmol/L cycloheximide. 相似文献
6.
U. K. Nadjimov M. S. Mirakhmedov B. U. Nasirullaev G. N. Fatkhullaeva I. M. Scott 《Journal of Plant Growth Regulation》1996,15(3):129-131
Fusicoccin (FC) was applied as a spray to shoots of intact field- and glasshouse-grown cotton plants. Distortions of shoot morphology resulted. Stems and petioles of FC-treated plants were irregular in diameter and twisted, whereas leaf laminae were curled and crinkled. Shoot elongation was inhibited by FC; the effect was dependent upon the concentration and timing of the applications.Abbreviation FC
fusicoccin 相似文献
7.
GhAGL15s,preferentially expressed during somatic embryogenesis,promote embryogenic callus formation in cotton (Gossypium hirsutum L.) 总被引:2,自引:0,他引:2
Zuoren Yang Changfeng Li Ye Wang Chaojun Zhang Zhixia Wu Xueyan Zhang Chuanliang Liu Fuguang Li 《Molecular genetics and genomics : MGG》2014,289(5):873-883
Somatic embryogenesis is a useful tool for gene transfer and propagation of plants. AGAMOUS-LIKE15 (AGL15) promotes somatic embryogenesis in many plant species. In this study, three homologous AGL15 genes were isolated from Gossypium hirsutum L., namely GhAGL15-1, GhAGL15-3, and GhAGL15-4. Their putative proteins contained a highly conserved MADS-box DNA-binding domain and a less conserved K domain. Phylogenetic analysis suggested that the three GhAGL15s clustered most closely with AGL15 proteins in other plants. Subcellular location analyses revealed that three GhAGL15s were localized in the nucleus. Furthermore, their expression levels increased following embryogenic callus induction, but sharply decreased during the embryoid stage. GhAGL15-1 and GhAGL15-3 were significantly induced by 2,4-D and kinetin, whereas GhAGL15-4 was only responsive to 2,4-D treatment. Over-expression of the three GhAGL15s in cotton callus improved callus quality and significantly increased the embryogenic callus formation rate, while GhAGL15-4 had the highest positive effect on the embryogenic callus formation rate (an increase from 38.1 to 65.2 %). These results suggest that over-expression of GhAGL15s enhances embryogenic potential of transgenic calli. Therefore, spatiotemporal manipulation of GhAGL15s expression may prove valuable in improving cotton transformation efficiency. 相似文献
8.
9.
John J. Finer Ann A. Reilley Roberta H. Smith 《In vitro cellular & developmental biology. Plant》1987,23(10):717-722
Summary Maintainable, highly embryogenic suspension cultures of a wild relative of cotton (Gossypium klotzschianum Anderss.) have been obtained. Callus with no apparent organization was used to establish the liquid culture. Callus growth
conditions as well as suspension medium composition were optimized. A visual selection scheme was beneficial for the maintenance
of the embryogenic suspension. These liquid cultures have been maintained for over 10 mo. with no loss in embryogenic capacity.
The somatic embryos developed after transfer of the embryogenic tissues to a hormone-free liquid medium.
Salaries and research support were provided by State and Federal funds appropriated to OSU-OARDC. This is journal article
No. 71-87. 相似文献
10.
Morphogenesis and regeneration from stomatal guard cell complexes of cotton (Gossypium hirsutum L.) 总被引:1,自引:0,他引:1
The development of a regeneration system from cotton stomatal guard cells directly on epidermal strips is described. The
most important factors affecting embryogenic callus initiation in both of the varieties tested (Coker 312 and 315) were the
source of the epidermal tissue, including plant age (4–5 months old), the developmental stage of the flower (opening flower
stage) from which bracts were obtained, the composition of the culture medium and light irradiance. The flower developmental
stage was critical for callus formation, which was observed only from bracts obtained from opening flowers. In addition, epidermal
strips excised from the bract basal region were more responsive in culture than those obtained from the top region. Improved
callus initiation was obtained on epidermal strips which had their cuticle in contact with the culture medium. Light irradiance
was a limiting factor for embryogenic callus formation, which was observed only in calluses cultured under the lower light
irradiance (15.8 μmol m–2 s–1). Somatic embryogenesis was observed on callus cultures subcultured consecutively to a culture medium containing naphthalene
acetic acid (10.7 μM) and isopentenyladenine (4.9 μM). Histodifferentiation of somatic embryos was improved on a medium containing naphthaleneacetic acid (8.1 μM)+isopentenyladenine (2.5 μM) and abscisic acid (0.19–0.38 μM). Somatic embryo germination and plantlet development were obtained using established protocols with few modifications. On
average, one fully developed plant was obtained from the culture of circa 100 epidermal strips in both cultivars.
Received: 19 May 2000 / Revision received: 25 August 2000 / Accepted: 29 August 2000 相似文献
11.
12.
John Ruyack Michael R. Downing Judy Su Chang Earl D. Mitchell Jr. 《In vitro cellular & developmental biology. Plant》1979,15(5):368-373
Summary In vitro conditions are defined for starting and maintaining callus and suspension, cells from two cotton (Gossypium hirsitum L.) varieties, Im 216 and Acala 44, which are resistant and susceptible, respectively, to the bacterial pathogenXanthomonas malvacearum (E. F. Sm.) Dows. A light, friable callus was easily obtained and has been maintained for over 4 years. Whether stems or
leaves, the explant source for callus initiation made no difference for growth of callus tissue. Acala 44 callus had a fresh-weight
doubling time of 4 to 5 days, and Im 216 callus had a fresh-weight doubling time of 4 to 9 days; however, in suspension culture
the fresh-weight doubling times for Im 216 and Acala 44 were 6 days. The pH of the suspension medium dropped to 4.7 during
the exponential growth phase and rose to 5.4 at the stationary phase. Attempts to induce root and shoot initiation from these
callus cells were unsuccessful; however, greening of the callus tissue did occur. Schenk and Hildebrandt medium was used for
both callus and suspension cultures. Inoculation of Im 216 and Acala 44 callus tissues with two races ofX. malvacearum resulted in a resistant and susceptible response, respectively.
This research was supported in part by C. S. R. S. Grant 315-16-96 and the Agricultural Experiment Station of Oklahoma State
University. 相似文献
13.
Embryogenic callus and suspension cultures of eastern white pine (Pinus strobus) have been obtained. The whole female gametophyte was plated on a medium containing 50 mg/l glutamine, 500 mg/l casein hydrolysate, 3% sucrose, 2 mg/1 2,4-D, 1 mg/1 BA and 0.2% Gelrite as a solidifying agent. Embryogenic calli could be seen as early as 5 days following culture. Histological studies indicate proliferation of pre-existing embryogenic tissue in the corrosion cavity followed by extrusion of embryogenic callus through the micropylar end of the gametophyte. Embryogenic suspension cultures were obtained by placing embryogenic callus into liquid medium. Embryogenic suspension cultures were subcultured weekly and proliferated as early-stage embryos with attached suspensors. Embryo development was obtained following transfer of the embryogenic tissue to an auxin-free medium containing 50 mM glutamine, 38 M abscisic acid, and 6% sucrose. Although embryo development could be consistently obtained, whole plants have not yet been recovered from these somatic embryos.Abbreviations 2,4-D
2,4-Dichlorophenoxyacetic acid
- ABA
Abscisic acid
- BA
6-Benzyladenine
Salaries and research support were provided by State and Federal funds appropriated to OSU/OARDC. Journal Article No. 62–89 相似文献
14.
Still P. E. Plata M. I. Campbell R. J. Bueno L. C. Chichester E. A. Niblett C. L. 《Plant Cell, Tissue and Organ Culture》1987,9(1):37-43
Highly morphogenic callus cultures were isolated from stamens of a wild peanut species, Arachis paraguariensis. These cultures were initiated on modified N6 medium containing 0.2 mg1l-1 4amino-3,5,6-trichloro-picolinic acid (picloram) and 0.5 mg l-1 6-benzylaminopurine (BAP) and were maintained on modified N6 medium with 0.008 mg l-1 picloram and 0.25 mg l-1 BAP. Buds formed on the calli growing on the maintenance medium developed into shoots when they were transferred to a MS salts based medium with no hormones. The cultures could also be maintained as a suspension culture in N6 liquid medium. When cell clumps larger tham 840 m were collected from the suspension culture and transferred to MS medium without hormones, they formed shoots in liquid culture. Root formation rarely occurred in agar or liquid cultures. Therefore, grafting to stems of rooted seedlings was used to obtain plants from regenerated shoots. Eight out of 50 field grown plants produced viable seed. 相似文献
15.
16.
Somatic embryogenesis from leaf and petiole callus cultures of Gossypium hirsutum L. 总被引:2,自引:0,他引:2
Leaf discs from four strains and petioles from six strains of Gossypium hirsutum were cultured on a variety of media. Callus formed from explants on all media, though embryogenesis was highly specific. Embryos formed from only three strain x media combinations. A small percentage of these embryos developed into plantlets. These results demonstrate that cotton plants can be obtained from leaf tissue explants.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- Kn
Kinetin
- NAA
1-naphthalene acetic acid
College of Agricultural Sciences Publication Number T-4-193; this research was partially funded by the USDA-ARS Plant Stress and Water Conservation Research Program 相似文献
17.
Kui Shin Voo Clayton L. Rugh Joe C. Kamalay 《In vitro cellular & developmental biology. Plant》1991,27(3):117-124
Summary We describe a tissue culture procedure for somatic embryogenesis and plantlet regeneration in cotton (Gossypium hirsutum L. cv. Coker 312). Callused explants or individual globular embryos were transferred to basal media to induce somatic embryogenesis.
To determine characteristic early indicators of successful germination and conversion, we identified six types of embryos
that developed on basal media. Two of the six embryo types, designated as tulip-shaped and trumpet-shaped, could undergo conversion
in preliminary tests, whereas the others had little or no developmental potential. Several media treatments designed to enhance
the maturation of globular somatic embryos failed to increase the fraction of embryos which matured to form recoverable types.
In efforts to improve plantlet recovery, tulip-shaped embryos were used in limited trials to contrast the effects of chemical
and physical desiccation treatments on germination and conversion. The selective use of tulip-shaped somatic embryos, coupled
with partial desiccation, seems to have augmented plant recovery. Growth habit, flowering, seed set, and lint production of
most of the regenerated plants were comparable to seed-derived plants grown under the same conditions.
Partial research support was provided by state and federal funds appropriated to the Ohio Agricultural Research and Development
Center, The Ohio State University. 相似文献
18.
Benjamin Steinitz Yedidya Gafni Yael Cohen Josefina Perea Diaz Yona Tabib Shlomit Levski Amos Navon 《In vitro cellular & developmental biology. Plant》2002,38(3):247-251
Summary The insecticidal effectiveness of a δ-endotoxin Cry protein from Bacillus thuringiensis in non-regenerable callus of a commercial Gossypium hirsutum L. variety was investigated. Two transgenic callus types were generated. The first callus type harbored the cry1A(c) gene and the hygromycin B phosphotransferase hpt selectable marker gene. The second callus type, the transgenic control, carried the marker genes β-glucuronidase (GUS) and
hpt. Growth and survival rates of three major cotton moth species, Pectinophora gossypiella, Helicoverpa armigera, and Spodoptera littoralis, were examined with aseptic neonates reared on callus. Normal larval development occurred in all species supplied with non-transgenic
callus, but insects died, or their growth was severely restricted, when reared on transgenic callus harvested from hygromycin
B-supplemented medium. Development of larvae on transgenic control and on non-transgenic callus became very much alike after
the transgenic control tissue had been subcultured on a hygromyein B-free medium for about 100 d prior to the insect-callus
bioassay. Accordingly, for detection of Bt toxin activity without the interference of the influence of hygromycin B on insects,
cry1A(c) callus was infested with insects after it had been propagated for more than 100 d on a medium free of the antibiotic. Under
these experimental conditions all P. gossypiella and H. armigera, and most S. littoralis neonates died, and the growth (e.g., weight increment) of S. littoralis survivors was markedly impeded by cry1A(c) callus. Three new findings emerge from this study: first, P. gossypiella, a pest feeding in the field on bolls only, can be grown in vitro on cotton callus; second, in a host which is recalcitrant in terms of plant regeneration, the biological potency of an insectdetrimental
transgene can nevertheless be evaluated by generating a transgenic host callus and conducting in vitro transgenic callus-insect assays; and third, our results suggest that hygromycin B is toxic to lepidopteran larvae. 相似文献
19.
Transformation of cotton (Gossypium hirsutum L.) by Agrobacterium tumefaciens and regeneration of transgenic plants 总被引:2,自引:0,他引:2
Ebrahim Firoozabady David L. DeBoer Donald J. Merlo Edward L. Halk Lorraine N. Amerson Kay E. Rashka Elizabeth E. Murray 《Plant molecular biology》1987,10(2):105-116
Cotton (Gossypium hirsutum L.) cotyledon tissues have been efficiently transformed and plants have been regenerated. Cotyledon pieces from 12-day-old aseptically germinated seedlings were inoculated with Agrobacterium tumefaciens strains containing avirulent Ti (tumor-inducing) plasmids with a chimeric gene encoding kanamycin resistance. After three days cocultivation, the cotyledon pieces were placed on a callus initiation medium containing kanamycin for selection. High frequencies of transformed kanamycin-resistant calli were produced, more than 80% of which were induced to form somatic embryos. Somatic embryos were germinated, and plants were regenerated and transferred to soil. Transformation was confirmed by opine production, kanamycin resistance, immunoassay, and DNA blot hybridization. This process for producing transgenic cotton plants facilitates transfer of genes of economic importance to cotton. 相似文献
20.
A rapid, clonal propagation procedure has been developed to regenerate mature cotton (Gossypium hirsutum L.) plants from pre-existing meristems that were excised from in-vitro-grown tissues. This plant regeneration procedure was
applicable to diverse cotton germplasms and required specific concentrations of 6-benzylaminopurine (BA) depending on the
origin of the meristems. All shoots regenerated directly without a callus phase. Screening BA concentrations (0.0–10.0 μm) demonstrated that shoot meristems (apices), secondary leaf nodes, primary leaf nodes, and cotyledonary nodes derived from
in-vitro-grown 28-day-old seedlings (Paymaster HS26) varied in their ability to form elongated shoots depending on the level
of BA. Indicative of a germplasm-independent procedure, a BA concentration screen (0.0, 0.3, 1.0 μm) demonstrated that explants with pre-existing meristems, excised from diverse germlines, were also able to form elongated
shoots at 0.3 μm BA. In most cases, elongated shoots derived from this procedure were rooted by a two-step process: an in-vitro maturation
step (Murashige and Skoog medium-activated charcoal) followed by planting into soil after basal application of Rootone. This
BA plant regeneration procedure was rapid, reproducible, and highly efficient for Stoneville 7A, Paymaster HS26, and other
high-fiber-yielding germlines. Regenerated plants were phenotypically normal and all of the mature plants regenerated to date
have initiated flowers and set viable R1 seeds.
Received: 15 March 1997 / Revision received: 28 August 1997 / Accepted: 5 September 1997 相似文献