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1.
Plant development from microspore-derived embryos in oilseed rape as affected by chilling, desiccation and cotyledon excision 总被引:1,自引:0,他引:1
The present study evaluated the effects of chilling, partial desiccation, cotyledon excision and successive subculture of
microspore-derived embryos on plant development in oilseed rape (Brassica napus L.). The results showed that out of the five media, all the genotypes showed the best response when the embryos were cultured
on the half-strength Murashige and Skoog medium with 2.0 mg dm−3 benzylaminopurine. A cold treatment for 3 or 5 d further increased frequencies of embryo germination (90.0 %) and plantlet
development (58.46 %). Desiccation for one day also increased the embryo germination and plantlet development in all genotypes
tested. Cutting the cotyledons of the embryos at late cotyledonary stage significantly increased the frequency of plantlet
development. The highest rate of plantlet development was obtained from cultures of embryos sampled with size of less than
4.0 mm. The successive subculture further improved the germination and development of plantlets from embryos. In the genotype
ZJU452, the rate of plantlet development reached 99.78 % after the second subculture of embryos. 相似文献
2.
J. Austin Burns Hazel Y. Wetzstein 《In vitro cellular & developmental biology. Plant》1995,31(2):72-78
Aggregates of globular and pre-globular stage somatic embryos from suspension cultures of pecan (Carya illinoensis Koch) were cultured on solidified media for embryo development. Embryo aggregates and pre-globular stage embryo masses were
given various treatments to further ontologic development. A 2- to 4-wk mild dehydration of the embryo aggregates suppressed
recurrent embryogenesis, promoted development of globular embryos into cotyledonary stage embryos, and enhanced plant development
beyond germination. Fine embryogenic tissue masses filtered from suspension formed cotyledonary-staged embryos when the collection
filters were plated on solified medium. The embryogenic capacity of preglobular stage embryo masses was compared between media
supplemented with varying concentrations of polyethylene glycol (molecular weight 8 000) vs. filter overlays. The filter paper
overlays were not necessary for embryo development. An inverse relationship was found between the number of embryos that developed
and the concentration of polyethylene glycol in the medium. However, this relationship was reversed for ability of embryos
to germinate and develop into a plant. 相似文献
3.
High frequency of plant regeneration in sunflower from cotyledons via somatic embryogenesis 总被引:7,自引:0,他引:7
Summary A plant regeneration methodvia somatic embryogenesis of severalHelianthus annuus L. genotypes was developed. Starting from cotyledonary explants high frequency embryo induction was obtained following several subcultures on defined media. An appropriate cotyledon developmental stage was identified. Etiolated explants and darkness treatment were necessary to obtain somatic embryos in all tested genotypes. After 20–25 days on somatic induction medium containing an auxin:cytokinin ratio of 1:1, the germination of embryos was induced by a reduction of the hormonal ratio (1:2). Shoots were excised from callus and transferred onto a medium containing various vitamins. The range of embryogenesis frequency was 33–72%, depending on the genotype. High frequency of rooting (49–82%) was obtained using a medium supplemented with 0.5 mg/L of ancymidol and by a reduction of photoperiod. A large percentage of somatic embryos developed into normal regenerated plants producing viable seeds.Abbreviations MS
Murashige and Skoog (1962)
- NAA
naphthaleneacetic acid
- BAP
benzylaminopurine
- GA3
gibberellic acid
- EIM
embryo induction medium
- GM
germination medium
- VM
vitamins medium
- RA2
ancymidol rooting medium
- EtOH
ethanol 相似文献
4.
Summary Factors influeneing the conversion of isolated mature zygotic embryos of Pinus radiata D. Don into plantlets were investigated. Nutritional factors were critical to this conversion. The optimum medium strength
was half-strength medium consisting of modified Quoirin and Le Poivre salts (von Arnold and Eriksson, 1981) and Schenk and
Hildebrandt (1972) vitamins. Sucrose (3%), glucose (2–3%) and fructose (2–5%) could serve as carbon sources for this conversion.
In general, a few significant benefits were found with the addition of organic nitrogen sources tested on the development
of isolated embryos into plantlets. Nearly all plant growth regulators tested were not beneficial for this conversion, and
some of them had a negative effect. Only gibberellic acid (GA3 at 0.58 μM) seemed to stimulate embryos to germinate a little bit earlier in comparison with the control. Submerging the cotyledons
of the isolated embryo into the agar-gelled medium led to better growth in comparison with the control. Embryos cultured in
liquid medium grew better but the germination percentage was apparently lower compared with agar-gelled medium. Liquid medium
with sponge support could increase the percentage of germinated isolated embryos, but the embryo growth was not comparable
to the liquid medium only. The addition of polyethylene glycol 6000 to the liquid medium seemed to increase the germination
percentage and had no adverse effect on the growth of isolated embryos. Light could influence embling growth in different
ways. 相似文献
5.
Hui Song Qun-Feng Lou Xiang-Dong Luo Joseph N. Wolukau Wei-Ping Diao Chun-Tao Qian Jin-Feng Chen 《Plant Cell, Tissue and Organ Culture》2007,90(3):245-254
Six experiments (including pretreatment, embryonic callus induction media, preculture conditions, embryo induction media,
embryo germination media, and genotypic effects) were conducted to develop an efficient cucumber (Cucumis sativus L., 2n = 2x = 14) anther culture protocol. Pretreatment and embryo induction were key factors for successful anther culture. Suitable
temperature stress depended on the ecotype, i.e., cucumbers from cold areas responded well to cold shock whereas those from
temperate areas responded well to heat treatment. The best medium for embryonic callus induction was MS medium supplemented
with 4.44 μM BA, 2.26 μM 2, 4-D, 4.64 μM KIN, 3% sucrose and 0.8% agar. For embryo induction, MS medium supplemented with
0.54 μM NAA, 13.32 μM BA, 3% sucrose and 0.8% agar was optimal, and for embryo germination MS medium containing 2.22 μM BA,
6% sucrose and 1.2% agar was best. Using this protocol, we produced callus from 16 genotypes and regenerated plants from three
of 20 evaluated. Three embryos per anther and 42 DH per 45 anthers (93% success) were obtained for cv. Ningjia No. 1, which
was an improved result over a previous report. The origin of regenerants from microspores was determined by cytological, morphological
and AFLP analyses. 相似文献
6.
Influence of external factors on secondary embryogenesis and germination in somatic embryos from leaves of Quercus suber 总被引:2,自引:0,他引:2
Bárbara Fernández-Guijarro Cristina Celestino Mariano Toribio 《Plant Cell, Tissue and Organ Culture》1995,41(2):99-106
Somatic embryogenesis was obtained in cultures of leaves from young seedlings of Quercus suber L. A two-stage process, in which benzyladenine and naphthaleneacetic acid were added first at high and then at low concentrations, was required to initiate the process. Somatic embryos arose when the explants were subsequently placed on medium lacking plant growth regulators. The embryogenic lines remained productive, by means of secondary embryogenesis, on medium without growth regulators. However, this repetitive induction was influenced by the macronutrient composition of the culture medium. Both low total nitrogen content and high reduced nitrogen concentration decreased the percentage of somatic embryos that showed secondary embryogenesis. Our results suggest that alternate culture on medium that increases embryo proliferation and a low salt medium prohibiting embryo formation will partially synchronize embryo development. Chilling slightly reduced secondary embryogenesis but gave a modest increase in germination. Maturation under light followed by storage at 4 °C for at least 30 days gave the best results in switching embryos from an embryogenic pathway to a germinative one. Under these conditions 15% of embryos showed coordinated root and shoot growth and 35% formed either shoots or mostly roots. These percentages were higher than those of embryos matured in darkness. This result indicates that a specific treatment is required after maturation and before chilling to activate the switch from secondary embryo formation to germination.Abbreviations BA
benzyladenine
- NAA
naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
indolebutyric acid
- MS
Murashige & Skoog (1962) medium
- SH
Schenk & Hildebrandt (1972) medium
- G
Gamborg (1966, PRL-4-C) medium (macronutrients in mg l–1: NaH2PO4·H2O, 90; Na2HPO4, 30; KCl, 300; (NH4)2SO4, 200; MgSO4·7H2O, 250; KNO3, 1000, CaCl2·2H2O, 150)
- PGR
plant growth regulator 相似文献
7.
A. H. McKently 《Plant Cell, Tissue and Organ Culture》1995,42(3):251-254
Genotypes representing the three botanical varieties of peanut (Arachis hypogaea L.) were assessed for somatic embryogenesis and subsequent plant conversion from mature zygotic embryo axes. Explants were initially cultured on Murashige and Skoog medium supplemented with 12.42 M 4-amino-3,5,6-trichloropicolinic acid. Individual somatic embryos wer isolated from explant tissue and used to initiate repetitive liquid cultures. There were significant differences among genotypes and varieties for somatic embryo formation and plant regeneration using a single media sequence. Botanical variety fastigiata had a lower embryogenic frequency and produced significantly fewer embryos than either hypogaea or vulgaris, which were similar in response.Abbreviations EA
zygotic embryo axes
- MS
Murashige and Skoog (1962) medium
- picloram
4-amino-3,5
- 6
trichloropicolinic acid 相似文献
8.
Marzena Warchoł Ilona Czyczyło-Mysza Izabela Marcińska Kinga Dziurka Angelika Noga Edyta Skrzypek 《Acta Physiologiae Plantarum》2018,40(5):93
Doubled haploid (DH) technology in oat has not reached the same stage as in other cereals leading to its application in plant breeding. The objective of this investigation was to increase the effectiveness of Avena sativa L. haploid embryo germination obtained by the distant crosses with maize. Developed embryos (obtained from 22 genotypes) were transferred on five germination media: MS (Murashige and Skoog, Physiol Plant 15:473–497, 1962) with 3% sucrose, pH 5.8 (control medium), and 190-2 supplemented with 6 and 9% maltose. The pH of 190-2 was adjusted to 5.5 and 6.0. Of all tested genotypes, 591 haploid embryos were obtained, almost half of them (279) germinated. The rate of haploid embryo germination induced on 190-2 was 6.92%, while in MS it was 3.25%. The sugar and its concentration significantly affected the germination of haploid embryos. The highest percentage of haploid embryo germination (9.11%) and DH lines production (1.64%) was achieved on 190-2 with 9% maltose and pH 6.0. All DH lines are incorporated to breeding programs for the development of new cultivars. 相似文献
9.
Fernando Pliego-Alfaro Richard E. Litz Pamela A. Moon Dennis J. Gray 《Plant Cell, Tissue and Organ Culture》1996,44(1):53-61
Development of cotyledonary-stage nucellar embryos of mango was arrestedin vitro by exposure to 750–1750 M ABA. The enlargement and germination of nucellar embryos was inhibited for as long as 4 weeks after subculture from ABA-containing medium. Mannitol at concentrations between 7.5 and 12.5% inhibited nucellar embryo development, presumably due to osmotic effects; however, there was no residual effect after subculture of somatic embryos onto medium without mannitol. Temperatures between 22.5 and 37.5°C stimulated embryo development, whereas lower temperatures (7.5 and 15°C) delayed germination. There was no germination 1 month after somatic embryos, pulsed for 8 weeks at 7.5°C, were transferred to 22.5°C; however, after 2 months, 86% of these somatic embryos germinated. These results indicate that it is possible to induce developmental arrest in recalcitrant mango embryos with high concentrations of ABA, mannitol or low temperature (7.5°C).Abbreviations ABA
Abscisic acid
- MM1
Mango maturation medium 相似文献
10.
Acidification of the Medium Associated with Normal and Fusicoccin-Induced Seed Germination 总被引:1,自引:0,他引:1
Fusicoccin, a toxin stimulating cell enlargement and inducing proton extrusion in various plant tissues, has been shown to replace kinetin, gibberellic acid and red light in breaking seed dormancy. It also removes the inhibitory effect of abscisic acid. The present data also show that the stimulating effect of fucicoccin on embryo growth of decoated radish (Raphanus sativus L.) and maize (Zea mays) seeds and on the development of maize embryos is accompanied by an early, significant acidification of the medium. Acidification of the medium is also observed when fusicoccin reverses the abscisic acid-induced inhibition of germination. These results support the hypothesis that the mode of action of fusicoccin in promoting germination involves, as in stimulation of cell enlargement, the activation at the cell membrane level of proton extrusion processes. The physiological significance of fusicoccin-induced release of protons at the onset of germination is discussed in comparison with the results concerning the mechanism of action of fusicoccin on cell enlargement in other plant materials. 相似文献
11.
Improved somatic embryogenesis in wheat by partial simulation of the in-ovulo oxygen,growth-regulator and desiccation environments 总被引:1,自引:0,他引:1
J. G. Carman 《Planta》1988,175(3):417-424
The effects of O2, growth-regulators and desiccation on callus growth and somatic embryo (embryoid) development were investigated in cultures of immature embryos of two lines of Triticum aestivum L. Callus and embryoid formation were induced on media that contained N6-furfurylamin-opurine (kinetin) and either 2,4-dichlorophenoxyacetic acid or 3,6-dichloro-o-anisic acid, either with or without abscisic acid (ABA). Cultures containing differentiated embryoids were then exposed to high concentrations of both ABA and indole-3-acetic acid, after which samples were desiccated to approx. 10% tissue moisture. Incubating cultures in 3.2 mmol·l-1 O2 (approx. 9%, low-O2) increased embryoid formation sixfold in one wheat line and nearly threefold in another. In the former line low-O2 caused the formation of mostly embryogenic callus. Low-O2 also decreased precocious germination of immature embryos, decreased callus growth, and improved development and viability of the resultant embryoids. Including 1.9 mol·l-1 ABA in the callus-induction medium reduced germination of immature embryos and reduced the incidence of embryoids with visible abnormalities. Despite the improved morphology, significantly fewer of the embryoids produced on ABA-containing medium germinated. Desiccation significantly enhanced germination of these embryoids as well as those produced on ABA-free medium.Abbreviations ABA
abscisic acid
- DPA
days post-anthesis
- dicamba
3,6-dichloro-o-anisic acid
- 2,4-D
2,4-dichlorophen-oxyacetic acid
- FW
fresh weight
- IAA
indole-3-acetic acid
- Kin
kinetin (N6-furfurylaminopurine)
- MS
Murashige and Skoog (1962) medium
Contribution of the Utah Agricultural Experiment Station, Utah State University, Logan, UT, Journal Paper No. 3565 相似文献
12.
Ganeshan Seedhabadee Båga Monica Harvey Bryan L. Rossnagel Brian G. Scoles Graham J. Chibbar Ravindra N. 《Plant Cell, Tissue and Organ Culture》2003,73(1):57-64
The in vitro plant regeneration frequencies for immature scutella, leaf-bases/apical meristems (LB/AM) and mature embryos of four commercially important barley genotypes were compared. Production of shoots from mature embryos or calluses of LB/AM incubated on media containing 1.0 or 2.0 mg l–1 6-benzylaminopurine (BA) were comparable to regeneration frequencies obtained for scutella-derived calluses of the same genotypes. Incubation of excised mature embryos and LB/AM on media containing the plant growth regulator, thidiazuron (TDZ), resulted in an increased shoot production. However, TDZ treatment did not stimulate plant regeneration from calluses derived from scutella or LB/AM. Shoots formed from TDZ-treated mature embryos and LB/AM were induced without a callus interphase and the in vitro culture system gave a three- to eight-fold higher regeneration frequency than recorded for scutella-derived calluses on BA medium. The simplicity and rapid development of shoots using the mature embryo system could potentially be used for the regeneration and genetic transformation of barley over alternative regeneration systems. 相似文献
13.
Androgenesis of wheat, rice and triticale was studied in isolated microspore culture. It is the first publication which studies microspore culture reaction of Hungarian rice varieties. The effect of different basic media, lack and absence of growth regulators in culture media were tested on important parameters of microspore culture. Direct embryogenesis was observed in microspore culture of wheat and triticale genotypes. In the case of rice, calli were induced in isolated rice microspore culture and haploid rice plantlets were regenerated via organogenesis.In wheat, the effect of basic media (W14, A2, CHB3, P4-m) was compared and among them the W14, and A2 had a superior effect on embryo production and albino and green plantlet regeneration. In rice the C, CHB3 and MSm media were tested in microspore culture and the significantly highest numbers of calli were achieved by using C and CHB3 media depending on the genotypes. The lack of exogenous growth regulators was observed in isolated microspore culture of triticale and rice. Growth regulator-free medium had a positive effect on embryo production and plant regeneration of triticale genotypes, whereas in rice microspore culture multicellular structures did not continue their division without growth regulators from the third week of microspore culture. Developing of microspore-origin calli was maintained by supplement of 2,4-D and Kinetin combination in the microspore culture medium. 相似文献
14.
The factors influencing somatic embryo maturation, high frequency somatic embryo germination, and plantlet formation were
studied in Terminalia chebula Retz. Maturation of somatic embryo were influenced by a number of factors such as in vitro culture passage, concentrations
of sucrose, levels of abscisic acid (ABA), basal media and media additive combinations. Maximum frequency of somatic embryo
maturation (57.22 ± 2.02), was obtained on MS medium supplemented with 50 g/l sucrose. Different factors such as strengths
of MS nutrients, plant growth regulators, media additives and their combinations controlling somatic embryo germination and
plantlet formation were studied. High frequency of germination and plantlet formation (58.80 ± 1.47) were achieved by subsequent
subculture of mature somatic embryos on MS medium containing 30 g/l sucrose and 0.5 mg/l benzyladenine (BA). However, although
duration of in vitro passage of the callus tissue was critical, contribution of the combinations of plant growth regulators
and media additives showed nugatory effect on somatic embryo maturation and germination as evident from variable responses. 相似文献
15.
Somatic embryogenesis in two Gossypium hirsutum genotypes on semi-solid versus liquid proliferation media 总被引:1,自引:0,他引:1
A comparison of semi-solid vs. liquid embryo proliferation media was made using two Gossypium hirsutum L. genotypes (Coker 312 and T25) and two callus initiation media. Sections of petioles from mature, flowering plants were cultured on two modified Murashige and Skoog media. Medium 1 included 4.0 mg l-1 NAA and 1.0 mg l-1 kinetin; medium 2 contained 0.1 mg l-1 2,4-D and 0.1 mg l-1 kinetin. After six weeks, callus was removed from each explant and divided in half. One callus portion was placed in liquid proliferation medium and the other on semi-solid (0.2% Gelrite) proliferation medium. Composition of proliferation medium was identical to that of initiation medium, except no growth regulators were added. Embryos were counted after eight weeks. The percentage of explants forming callus was influenced by genotype/initiation medium combination. Analysis of variance procedures revealed significant variability for callus initiation media, proliferation media (semi-solid or liquid), and an initiation medium x genotype interaction. Paired t-tests indicated that more embryos were produced in liquid proliferation medium (227.3 embryos/culture) than on semi-solid proliferation medium (134.6 embryos/culture).Abbreviations NAA
naphtaleneacetic acid
- 2,4-D
2,4-D dichlorophenoxyacetic acid 相似文献
16.
S. Hindson A. R. McElroy C. Portelance 《In vitro cellular & developmental biology. Plant》1998,34(3):181-184
Summary Various preconditioning treatments of alfalfa (Medicago sativa L.) somatic embryos to improve embryo quality and conversion were studied. Four different regenerating genotypes were compared.
Embryogenic cultures were established in liquid culture. Globular embryos were collected and plated on an embryo development
medium until they reached cotyledonary stage. They were then exposed to three treatments: a standard embryo development medium
(control), media supplementation with 1 μM abscisic acid (ABA), 50 mM glutamine and 5% sucrose (T), additional supplementation with 50 μM ABA (TT), and additional supplementation followed by desiccation (TTD). Treatments affected embryo conversion, but not uniformly
for all genotypes. Embryo conversion was increased (P<0.05) by pretreatment (T), while only one exhibited any response to additional ABA (T vs. TT). Desiccation decreased (P<0.05) conversion of pretreated embryos (TT vs. TTD) of all genotypes. The effect of treatments on plantlet weight was less
pronounced and inconsistent across genotypes. 相似文献
17.
Anna Nadolska-Orczyk 《Plant Cell, Tissue and Organ Culture》1992,28(1):19-25
Somatic embryos were obtained from immature cotyledons of Lupinus angustifolius, L. albus and L. mutabilis but not from L. luteus. Different kinds of basal media and plant growth regulators in primary and secondary culture were tested. The best induction media were based on B5 and were supplemented with 5 mg I-1 2,4-D alone or with 0.25 mg I-1 kinetin. Mature stage somatic embryos were obtained on media containing ABA (0.1–0.5 mg I-1) and a high NH4/NO3 ratio. Embryo germination and plantlet development occurred on MS media supplemented with glutamine or GA3. 相似文献
18.
Gloria Pinto Yill-Sung Park Sónia Silva Lucinda Neves Clara Araújo Conceição Santos 《Plant Cell, Tissue and Organ Culture》2008,95(1):69-78
The described protocol for repetitive somatic embryogenesis (SE) in Eucalyptus globulus produced more somatic embryos than the primary SE protocol. Primary somatic embryos (induced on MS3NAA) were transferred to the same medium, leading to new cycles of somatic embryos, for at least 2 years. The influence of medium
(MS and B5), plant growth regulators (auxins and cytokinins), and light on secondary SE was tested. The MS medium without
growth regulators (MSWH) was more efficient for cotyledonary embryo formation and germination than the B5 medium. Reducing auxin (NAA) levels increased
the proliferation of globular somatic embryos and allowed SE competence to be maintained on medium free of plant growth regulators.
The addition of two cytokinins (BAP and KIN) to the MS medium did not improve proliferation of globular secondary embryos,
but was crucial during later stages of the SE process (germination and conversion). Data also show that light may influence
the quality of the process, depending on its stage. Darkness should be maintained until the cotyledonary stage is reached,
after which exposure to light is recommended. 相似文献
19.
We describe a protocol for somatic embryogenesis of Protea cynaroides, with potential for high frequency production of this important horticultural species. Somatic embryos formed directly on
both P. cynaroides mature zygotic embryos and excised cotyledons cultured on MS medium without growth regulators. The addition of growth regulators
such as naphthalene acetic acid (NAA) (5; 13 and 27 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (5; 11 and 23 μM), in combination
with thidiazuron (TDZ) (1 μM), benzylaminopurine (BAP) (1 μM) or kinetin (1 μM) suppressed the formation of somatic embryos.
After eight weeks in culture, formation of somatic embryos was observed. Zygotic explants formed the most embryos when cultured
in a 12-h photoperiod in comparison to explants cultured in the dark. Up to 83% of these embryos germinated after transferal
to the germination medium containing 0.3 μM GA3. Significantly fewer embryos germinated in MS medium with no growth regulators, or supplemented with higher concentrations
of GA3, while low germination percentages were also observed in MS media containing casein hydrolysate and coconut water. The germination
of normal somatic embryos (two separate cotyledons and a single radicle) was observed only in media containing either no growth
regulators, 0.3 μM GA3 or 1 μM GA3. All embryos that germinated in high concentrations of GA3 were malformed. 相似文献
20.
Dong-Ho Shin Jin Sook Kim In Jeong Kim Jaemo Yang Soo Kyung Oh Gab Chae Chung Kyung-Hwan Han 《In vitro cellular & developmental biology. Plant》2000,36(4):273-278
Summary We describe a protocol, and several experiments that helped lead to its development, for sunflower regeneration. Important
factors for sunflower regeneration were explant age, cytokinin type and concentration, basal medium, and explant source. We
could not induce shoot regeneration from the explants derived from mature tissues including leaf, petiole, and stem. However,
use of juvenile explants such as embryo meristem and primordial leaf tissues allowed routine regeneration of 17 different
sunflower genotypes. High frequency of shoot regeneration was achieved with these explants taken from seedlings up to 5 d
after germination. Explant age was less critical for embryo meristem explants than for primordial leaf tissues. Of the four
basal media tested, MS and B5 media produced higher shoot-regeneration frequencies than did Anderson and woody plant media.
The highest shoot-regeneration frequency was obtained with MS medium supplemented with 2 μM BA and without auxin. Addition of 1 μM naphthalene-acetic acid to the medium significantly reduced both the percentage of explants producing shoots and average
number of shoots per explant. Regenerated shoots were grown to maturity in a greenhouse. 相似文献