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1.
A method to produce encapsulatable units for synthetic seeds was developed in Asparagus officinalis L. Encapsulatable units with high conversion ability in non-sterile soil were produced from somatic embryos by a pre-encapsulation culture. The synthetic seeds containing somatic embryos without the pre-encapsulation culture did not germinate in soil. When the pre-encapsulation culture medium did not contain growth regulators, the roots elongated too much to accomplish encapsulation. Several growth regulators were studied and indole-3-acetic acid was considered to be optimum at 28.5 M. The pre-encapsulation culture medium with indole-3-acetic acid inhibited the growth of roots during the pre-encapsulation culture and produced compact encapsulatable units. The growth of roots was promoted when plants were produced from the encapsulatable units. The percent conversion of the synthetic seeds with these encapsulatable units was 72% in non-sterile soil. This is the first report on synthetic seeds in Asparagus officinalis L. 相似文献
2.
We have developed an electroporation procedure for the transformation of carrot protoplasts with Ti-plasmid DNA from Agrobacterium tumefaciens. The uptake of pTiC58 into carrot protoplasts was mediated by high voltage electrical pulses at field strengths from 0.5 to 3.8 kV/cm. Protoplast regeneration, somatic embryogenesis and plantlet regeneration were unaffected by the electroporation conditions selected for DNA uptake. Uptake of plasmid pTiC58 resulted in hormone independent regeneration of carrot protoplasts. Transformed somatic embryos were detected in carrot cultures 45 days after electroporation. The transformed somatic embryos developed into teratomas which synthesized nopaline. Hybridization was obtained between a labeled T-DNA fragment from pTiC58 and DNA fragments from 4 month old teratomas regenerated from electro-transformed protoplasts. Based on the number of somatic embryos regenerated after electro-transformation, a frequency of 1.6×102 transformants/104 somatic embryos/g pTiC58 DNA was obtained.Abbreviations PEG
polyethylene glycol
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MES
morpholinoethane sulfonic acid
- DMSO
dimethyl sulfoxide
- HSV
Herpes Simplex virus
- TK
thymidine kinase 相似文献
3.
Summary Somatic embryos of carrot were encapsulated in calcium alginate beads to provide artificial carrot seeds. Alginate capsules with a hardness of 0.2 to 0.5 kg/cm2 were found to be suitable for germination of encapsulated somatic embryos. The germination frequencies were more than 95%, when grown aseptically on polyester fiber supports loaded with hormone-free Murashige-Skoog medium. 相似文献
4.
A novel technique for the encapsulation of plant material in calcium alginate hollow beads was tested. The technique involves
suspending plant material (i.e. plant cells, tissues, organs, shoot tips, somatic embryos) in a solution containing carboxymethylcellulose
and calcium chloride and then dripping it into a stirred sodium alginate solution. In initial experiments with Daucus carota (carrot), it was found that after 14 days of cultivation, 100 % of seeds encapsulated in calcium alginate hollow beads would
germinate in the liquid core and that 13% would burst the capsules. Embryogenic calli developed inside hollow beads and formed
somatic embryos while calli in conventional calcium alginate beads became detached from the beads early in development, and
no somatic embryogenesis occurred. With Solanum tuberosum (potato), development of calli was observed in 50% of hollow beads. Eighty-one percent of shoot tips encapsulated in hollow
beads sprouted and grew out of the capsules.
Received: 28 October 1999 / Revision received: 11 February 2000 / Accepted: 22 February 2000 相似文献
5.
胡萝卜人工种子基本制作流程的建立 总被引:6,自引:0,他引:6
李修庆 《Acta Botanica Sinica》1989,31(9):673-677
通过在胚状体的产生、包埋和发芽过程中进行优化试验,建立胡萝卜人工种子基本制作流程,使人工种子在无菌条件下成苗率达96%,移栽到温室后成活率达79%。 相似文献
6.
Jean-Paul Ducos Hélène Bollon Vincent Pettard 《Applied microbiology and biotechnology》1993,39(4-5):465-470
Somatic carrot embryos were grown as batch cultures in a stirred 10-l bioreactor. Embryo production in the bioreactor was comparable to that obtained in shake-flasks. A production of about 50·103 embryos/l per day was commonly achieved with an inoculum density of 0.1% volume of tissues/volume of medium. Regularly changing of the medium increased embryo viability. A filtering unit coupled to the bioreactor was developed in order to calibrate embryos. The characteristics of the population of harvested embryos are described.
Correspondence to: J.-P. Ducos 相似文献
7.
Enhancing carrot somatic embryos survival during slow dehydration, by encapsulation and control of dehydration 总被引:3,自引:0,他引:3
In order to obtain dry artificial seeds, carrot somatic embryos were encapsulated and dehydrated. Encapsulation in some hydrogels delayed the dehydration and preserved the water content of carrot somatic embryos. In particular, a matrix made of alginate with gellan gum was found to be the most efficient in maintaining a high water activity (aw) around somatic embryos. By delaying dehydration, and also rehydration, encapsulation seemed to protect somatic embryos against desiccation and imbibition damages, giving better germination and emergence of cotyledons. Matrices made of alginate mixed with kaolin or gellan gum were particularly adapted to protect the embryos during the dehydration. Apart from the matrix composition, the control of dehydration speed enhanced the survival and regeneration of encapsulated-dehydrated somatic embryos. Using a slow dehydration protocol (95-15% RH—relative humidity into the chamber—in 11.5 days), it was possible to exert different dehydration speeds. Slowing the dehydration between 70 and 45% RH stabilized the water activity (aw) of the encapsulation matrix, and enhanced the survival and regeneration frequencies of encapsulated-dehydrated embryos. In the absence of any maturing pretreatment, alginate-gellan gum encapsulated carrot somatic embryos, dehydrated to 15% RH, and rehydrated in moistured air (90% RH), germinated up to 72.9%. Therefore, encapsulation in alginate-gellan gum, combined with a slow dehydration, leads to enhance the somatic embryos' desiccation tolerance. 相似文献
8.
Lenka Andrýsková Vilém Reinöhl Marek Klemš Stanislav Procházka 《Acta Physiologiae Plantarum》2009,31(4):675-681
Cucumber (Cucumis sativus L.) cytokinin-independent embryogenic cell suspension cultures were derived and maintained for more than 3.5 years without
losing the embryogenic potential. The preparation and the characteristics of the cucumber embryogenic cell suspension possess
many similarities to that of carrot. The cultures were induced from hypocotyl explants of in vitro grown cucumber plants in
liquid MS media containing 2,4-dichlorophenoxyacetic acid as the sole growth regulator during 6 weeks and they contained a
heterogeneous array of several different types of single cells and cell clusters (PEMs). The established cell suspensions
were subcultured in 1-week interval, while the inoculation density was optimized to 2.0 × 105 cells ml−1 using cell viability as a marker. Somatic embryos were obtained after the transfer of the proembryogenic masses to a hormone-free
semisolid MS medium with a frequency of 388 ± 57 somatic embryos per 1 ml of packed cell volume of the established cucumber
embryogenic culture within 7 days. The frequency of normal somatic embryos with two cotyledons was found to be 78%. Such embryos
possessed the potential of spontaneous maturation and the embryo conversion rates were 87%. The yield of normally growing
plants was much higher compared with that previously described for cucumber systems. Somatic embryo-derived plants were successfully
transferred to the greenhouse where they flowered and fruited. 相似文献
9.
Several plant growth regulators BA, TDZ, 2,4- and Kn were tested alone or in combination for their capacity to induce indirect somatic embryogenesis from leaf and internode explants of Paulownia elongata. Calli were produced when leaf explants were cultured on Murashige and Skoog (MS) medium containing 3 % sucrose, 0.4 % phytagel, 4 mg l-1 TDZ and 0.1 mg l-1 Kn after 3 weeks and the initiation rate was 54.1%. After subculturing on the same medium, embryos at various developmental stages (globular, heart and torpedo shaped) were transferred for maturation onto MS medium supplemented with 3 % sucrose, 0.4 % phytagel, 0.1 mg l-1 TDZ, 1 mg l-1 Kn and 2 mM glutamine. An average of 50.7 somatic embryos were obtained from 100 mg of embryogenic callus after 4 weeks at high frequency (64.7 %). Afterward, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination (80 %) and development into plantlets. Plantlets were transferred to pots with a mixture of peat and perlite in a 3:1 ratio and showed a survival rate of 70–80 %. Plantlets regenerated by this procedure were morphologically identical to the donor material and developed normally in the greenhouse. 相似文献
10.
Plant regeneration from protoplasts of embryogenic cell suspensions of Coffea arabica L. cv. caturra
Coffee plants were regenerated from protoplasts isolated from embryogenic cell suspension cultures derived from somatic embryos of Coffea arabica L. cv. caturra. Yields of viable protoplasts ranged from 1×105 to 6×105 protoplast/g fresh weight. Protoplast preparations usually contained no contaminating cells, and when present, the number of cells never exceeded 0.1% of the total. Plating efficiencies of protoplast ranged from 1 to 10%. Embryogenic protocolonies obtained after several subcultures in a medium supplemented with 0.5 mg/l each of benzylaminopurine, 2,4-dichlorophenoxyacetic acid and naphtaleneacetic acid, were transferred to a medium lacking plant growth regulators. Well differentiated embryos were formed in selected protocolonies that contained many embryos-like structures. Approximately 70% of the somatic embryos developed into green rooted plantlets which were succesfully transferred to vessels containing sterilized scoria. Plants grown for two months in scoria were finally transferred to greenhouse.Abbreviations B5
medium according to Gamborg et. al.(1968)
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphtaleneacetic acid 相似文献
11.
Several plant growth regulators BA, TDZ, 2,4- and Kn were tested alone or in combination for their capacity to induce indirect somatic embryogenesis from leaf and internode explants of Paulownia elongata. Calli were produced when leaf explants were cultured on Murashige and Skoog (MS) medium containing 3 % sucrose, 0.4 % phytagel, 4 mg l-1 TDZ and 0.1 mg l-1 Kn after 3 weeks and the initiation rate was 54.1%. After subculturing on the same medium, embryos at various developmental stages (globular, heart and torpedo shaped) were transferred for maturation onto MS medium supplemented with 3 % sucrose, 0.4 % phytagel, 0.1 mg l-1 TDZ, 1 mg l-1 Kn and 2 mM glutamine. An average of 50.7 somatic embryos were obtained from 100 mg of embryogenic callus after 4 weeks at high frequency (64.7 %). Afterward, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination (80 %) and development into plantlets. Plantlets were transferred to pots with a mixture of peat and perlite in a 3:1 ratio and showed a survival rate of 70–80 %. Plantlets regenerated by this procedure were morphologically identical to the donor material and developed normally in the greenhouse. 相似文献
12.
William R. Krul 《Plant Cell, Tissue and Organ Culture》1993,32(3):271-276
Roots of nine cultivars of carrot (Daucus carota L.) were exposed to one, two or three months cold treatment and cells isolated from cold-treated and control roots were assayed for the production of somatic embryos. Cells obtained from the one- or two-months cold treatments formed embryos earlier, produced embryos over a longer period of time and produced more embryos per callus than the controls. In contrast, cells obtained from roots exposed to three months cold displayed a reduction in all parameters of embryogenic competence and for some cultivars this treatment resulted in production of cells with no embryogenic competence. The relationship of cold treatment of stock plants to the induction of metastable and stable patterns of gene expression and the induction of somatic embryos are discussed. 相似文献
13.
de Feria Manuel Jiménez Elio Barbón Raúl Capote Alina Chávez Maité Quiala Elisa 《Plant Cell, Tissue and Organ Culture》2003,72(1):1-6
The effect of two different dissolved oxygen (DO) concentrations (50 and 80%) on differentiation of somatic embryos (SE) from cell suspensions of coffee (Coffea arabica cv. Catimor 9722) was analyzed. Two bioreactors CMF-100 (CHEMAP AG) designed for the culture of cells, with 2-l glass vessels and a maximum work volume of 1.8 l were used. Each one was equipped with a gas blending unit (air, O2, N2, CO2) for the control of DO concentration. The inoculation density of embryogenic cells was 1.0 gram of fresh weight per liter (g FW l–1). The number of somatic embryos was greater (71 072 SE l–1) with 80% DO, but the major proportion were globular and heart shaped SE (66 399 SE l–1) and only 6.6% with regard to total was torpedo shaped SE. However, the 50% DO produced the higher number in the torpedo shaped SE (7389 SE l–1) what represented 20.0% with regard to total. Thus, higher concentrations of DO induced globular and heart shaped SE differentiation, but for production of torpedo shaped SE lower concentrations DO are needed. The somatic embryos obtained in the bioreactor with 50% DO showed similar behavior to the somatic embryos obtained in the rotary shaker. After 8 weeks of culture, 49.2% germination was obtained, which allowed a total of 1725 plantlet to be transferred to conditions ex vitro. After 6 months of culture, 89.2% of conversion was achieved and 1539 plants obtained were transferred to field conditions. 相似文献
14.
Summary A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l–1 kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l–1 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.Abbreviations MS
Murashige and Skoog (1962)
- 2,4-D
2,4 dichlorophenoxyacetic acid
- BA
6-benzyladenine 相似文献
15.
Summary An efficient culture system has been developed for repeated cycles of somatic embryogenesis in microspore-derived embryos of Brassica juncea without a callus phase. Haploid embryos produced through anther culture showed a high propensity for direct production of somatic embryos in response to 2 mgL–1 BA and 0.1 mgL–1 NAA. The embryogenic cultures which comprised the elongated embryonal axis of microspore-derived embryos when explanted and grown on the medium of same composition produced a large number of secondary embryos. These somatic embryos in turn underwent axis elongation and produced more somatic embryos when explanted and cultured. This cycle of repetitive somatic embryogenesis continued with undiminished vigour passage after passage and was monitored for more than a year. Somatic embryos from any passage when isolated at cotyledonary stage and grown on auxin-free medium for 5 days and then on a medium containing NAA (0.1 mgL–1), developed into complete plants with a profuse root system and were easily established in the soil. The cytology of the root tips of these plants confirmed their haploid nature. The total absence of callus phase makes the system ideal for continuous cloning of androgenic lines, Agrobacterium-mediated transformation and mutation induction studies. 相似文献
16.
A. Nordine C. R. Tlemcani A. El Meskaoui 《In vitro cellular & developmental biology. Plant》2014,50(1):19-25
A protocol for somatic embryogenesis was developed for Thymus hyemalis, a wild species in the Mediterranean region. First, the effects of explant type, plant growth regulators [kinetin (KIN) and 2,4-dichlorophenoxyacetic acid (2,4-D)], and genotype on callus induction were tested. For callus induction, the node was the best explant; Murashige and Skoog (MS) medium supplemented with 1.8 μM 2,4-D and 0.5 μM KIN was the best medium, and the genotype had a highly significant effect. To induce production of somatic embryos, the effects of KIN, 6-benzylaminopurine (BAP), and naphthalene acetic acid (NAA) were evaluated. After 5 wk of culture in the dark, MS medium supplemented with 4.44 μM BAP, 0.54 μM NAA, and 4.65 μM KIN gave the highest percentage (85%) of embryogenic callus and the highest number of somatic embryos (27.00) per 45 mg of callus. For germination and plant recovery, somatic embryos were transferred to MS medium without plant growth regulators and plantlet conversion from developed somatic embryos was 90%. In vitro plants with adequate growth and sufficient root systems were subsequently transplanted into a mixture of peat and vermiculite (2:1?v/v) under greenhouse conditions. The survival rate of the plantlets under ex vitro conditions was 80%. 相似文献
17.
Silvia Karina Vila Hebe Yolanda Rey Luis Amado Mroginski 《Journal of Plant Growth Regulation》2007,26(3):268-277
Factors affecting somatic embryogenesis induction and conversion in paradise tree (Melia azedarach) were evaluated. Somatic embryogenesis was influenced by plant growth regulators, explant stage, carbohydrate source and
concentration, gelling agents, light, and induction times. MS medium with 4.54 μM thidiazuron (TDZ) was optimal for the induction
of embryogenic tissue. Zygotic embryos that were 1-1.5 mm long (torpedo and early cotyledonal stage) had a greater embryogenic
response than smaller or larger embryos and better conversion of somatic embryos into plants. In general, embryos that formed
in medium containing 1% or 5% carbohydrate were hyperhydrics or fused, respectively, whereas those that formed in medium with
a carbohydrate concentration of 3% had better morphology. Raffinose at 3% yielded satisfactory somatic embryo induction with
good morphology and the best values of conversion into plants. Induction and conversion of somatic embryos were superior on
medium solidified with agar A-1296. The explants maintained under 160 μmol m−2 s−1 or 1 week in darkness and later 160 μmol m−2 s−1 produced a significantly higher embryogenic index. Only 4 days of treatment on induction medium, with either raffinose or
sucrose at 3% as a carbohydrate source, were required to induce somatic embryogenesis, but longer exposure, until 18 days,
increased the yield and improved the morphology of somatic embryos. 相似文献
18.
Summary
In vitro propagation of Quassia amara L. (Simaroubaceae) was attempted using mature and juvenile explants. Attempts to establish in vitro culture using leaf and internode explants from a plant more than 15yr old were unsuccessful due to severe phenolic exudation.
Plant regeneration through direct and indirect somatic embryogenesis was established from cotyledon explants. Murashige and
Skoog (MS) medium with 8.9 μM N6-benzyladenine (BA) and 11.7 μM silver nitrate induced the highest number (mean of 32.4 embryos per cotyledon) of somatic embryos. Direct somatic embryogenesis
as well as callus formation was observed on medium with BA (8.9–13.3 μM). Semi-mature pale green cotyledons were superior for the induction of somatic embryos. Embryos developed from the adaxial
side as well as from the point of excision of the embryonic axis. More embryos were developed on the proximal end compared
to mid and distal regions of the cotyledons. Subculture of callus (developed along with the somatic embryos on medium with
BA alone) onto medium containing 8.9 μM BA and 11.7 μM silver nitrate produced a mean of 17.1 somatic embryos. Primary somatic embryos cultured on MS medium with 8.9 μM BA and 11.7μM silver nitrate produced a mean of 9.4 secondary somatic embryos. Most of the embryos developed up to early cotyledonary stage.
Reduced concentration of BA (2.2 or 4.4 μM) improved maturation and conversion of embryos to plantlets. Ninety percent of the embryos converted to plantlets. The optimized
protocol facilitated recovery of 30 plantlets per cotyledon explant within 80d. Plantlets transferred to small cups were subsequently
transferred to field conditions with a survival rate of 90%. 相似文献
19.
Pieces of an embryogenic mass (EMS) induced in culture from immature fruits of pistachio, Pistacia vera L., were encapsulated into calcium alginate beads. Somatic embryos were also encapsulated individually into calcium alginate beads to produce synthetic seeds. The viability of the encapsulated EMS and somatic embryos was investigated immediately following encapsulation, and after storage for 60 days at 4°C. The encapsulated-stored EMS fragments recovered their original proliferative capacity after two months storage following two sub-cultures, but non-encapsulated-stored EMS failed to recover. The conversion frequency of synthetic seeds to seedling plants was 14% after storage for 60 days at 4°C, from which it may be concluded that encapsulation is a practical procedure for short-term storage of embryogenic pistachio tissue, and may be applicable to the preservation of desirable elite genotypes.Abbreviations BAP
Benzylaminopurine
- EMS(es)
Embryogenic mass(es)
- MS
Murashige and Skoog medium (Sigma M-0404)
- PGR(s)
Plant growth regulator(s) 相似文献
20.
Tomohiro Kiyosue Jiro Nakayama Shinobu Satoh Akira Isogai Akinori Suzuki Hiroshi Kamada Hiroshi Harada 《Planta》1992,186(3):337-342
ECP31, an embryogenic-cell protein from carrot (Daucus carota L.), was purified by sequential column-chromatographic steps and digested by V8 protease on a nitrocellulose membrane. The resultant peptides were separated by reverse-phased column chromatography and sequenced. The sequences obtained were 70–80% homologous to those of a late-embryogenesis-abundant protein (D34) from cotton (Baker et al, 1988, Plant Mol. Biol. 11, 227–291). The level of ECP31 in somatic embryos of carrot was increased by treatment of the embryos with 3.7 · 10–6 M abscisic acid (ABA) for 48 h, and there was no change in this enhanced level for up to 192 h in the presence of ABA. No similar enhancing effect of ABA was observed on the level of ECP31 in embryogenic callus or segments of carrot hypocotyls. In an immunohistochemical analysis, ECP31 was found in epidermal tissue and in the vascular system of ABA-treated somatic embryos.Abbreviations ABA
abscisic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- LEA protein
late-embryogenesis-abundant protein
To whom correspondence should be addressedThis work was supported in part by a grant-in-aid for Special Research in Priority Areas (Project No. 02242102) from the Ministry of Education, Science and Culture, Japan, and by Special Coordination Funds of the Science and Technology Agency of the Japanese Government. 相似文献