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1.
Immature zygotic embryos at different developmental stages were used for callus induction and regeneration studies. Immature embryos excised from fruits 77, 91, 100, 114, 128, 140 and 193 days after pollination and mature embryos were cultured on modified Y3 medium containing 500 mgl–1 cysteine, 0.5% (w/v) PVP-40, 500 M 2,4-d and 0.3% (w/v) charcoal. Compact embryogenic tissue began differentiating directly from embryo explants after 2 weeks of culture. The percentage of embryos forming compact embryogenic tissue ranged from 28.6% for 91-day-old embryos to 0% for 140-day-old and older embryos. Friable embryogenic tissue was observed in callus cultures derived from 100-day-old embryos. Although both compact and friable embryogenic tissues were successfully isolated, normal embryo and plantlet development was observed only from friable embryogenic tissue.Abbreviations ABA
abscisic acid
- 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- PVP
polyvinylpyrollidone 相似文献
2.
Somatic embryogenesis in peanut (Arachis hypogaea L.) using immature cotyledonary explants was induced on a wide range of 2,4-dichlorophenoxyacetic acid (2,4-D) (5 to 60mg l–1) and naphthaleneacetic acid (NAA) (20 to 50 mg l–1) levels. Percent embryogenesis ranged from 31 to 94%. As auxin level increased in induction medium, percent embryogenesis decreased and was associated with browning of explants. However, with higher 2,4-D induction levels (40 mg l–1 and over), embryogenic explants had dense masses of embryogenic areas and repetitive embryogenesis was enhanced. Higher auxin concentrations during induction decreased precocious germination of embryos, but had no marked effect on somatic embryo morphology. The use of 2,4-D compared to NAA in the induction medium resulted in greater per cent embryogenesis and mean number of embryos. Embryos induced on NAA were harder, less pliant, and less succulent; cultures exhibited more extensive root development and nonembryogenic callus proliferation.Abbreviations B5
Gamborg et al. (1968)
- BA
benzyladenine
- 2,4-D
dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- MS
Murashige & Skoog (1962)
- NAA
naphthaleneacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
3.
The objective of this study was to determine the types and concentrations of auxins and cytokinins that would result in effective culture initiation and plant regeneration of switchgrass (Panicum virgatum L.). Whole caryopses and young seedling explants were cultured on MS medium containing different concentrations of picloram or 2,4-dichlorophenoxyacetic acid (2,4-d) in combination with benzyladenine. Best results from mature caryopses were obtained with 11.3–45.0 M 2,4-d in combination with 15.0 or 45.0 M benzyladenine (BA). More than 300 plants were obtained per embryogenic callus from some treatments after the second transfer to regeneration medium (90 days after initiation of the cultures). Regeneration was obtained from young seedling explants with both auxins. However, picloram was more effective over a wider range of BA concentrations than 2,4-d. Protocols developed during this study were used to regenerate hundreds of plants which could easily be established in the field. 相似文献
4.
Zhao Yan-Xiu Yao Dun-Yi Philip J. C. Harris 《Plant Cell, Tissue and Organ Culture》1993,34(3):253-260
Root, hypocotyl and cotyledon explants of Sesbania bispinosa, Sesbania cannabina, Sesbania formosa, and Sesbania sesban were cultured on Murashige and Skoog medium with benzyladenine (BA; 2.22, 4.44, 8.88 M) in combination with 2,4-dichlorophenoxyacetic acid (2,4-d; 2.26, 4.52, 9.05 M), indolebutyric acid (IBA; 0.25, 0.49, 4.92 M) or naphthaleneacetic acid (NAA; 2.69, 5.37, 10.74 M). Although all explant types developed some callus, callus occurred earliest and continued to grow fastest with hypocotyls. Media including 2.4-d or NAA gave the fastest growing callus. Callus was subcultured up to 10 times at 20-day intervals and retained a rapid growth rate. Shoots regenerated readily from both hypocotyls or cotyledons but not from roots. Shoot organogenesis was most frequent with IBA (0.25–4.92 M) in combination with BA (4.44–8.88 M) and did not occur with 2,4-d. With each species at least one medium induced shoot differentiation from more than 50 percent of the callus pieces. With one exception, media containing IBA that induced shoot organogenesis on explants also did so in callus, but media containing NAA, even when effective with explants, did not cause differentiation of callus. Shoots that differentiated were excised and cultured on MS medium without growth regulators or with IBA (2.46, 4.92, 9.84 M). Roots developed after 3–8 days on an appropriate rooting medium, often without IBA. Rooted plantlets were transplanted to pots in a greenhouse and developed into normal plants. Suitable media and protocols for initiating and subculturing callus and regenerating whole plants in vitro from callus and explants have thus been established for four species of Sesbania. 相似文献
5.
Shoot tips of wheat as an alternative source for regenerable embryogenic callus cultures 总被引:5,自引:0,他引:5
Shoot tips of Triticum aestivum L. cvs. Turbo and Nandu, both summer wheat varieties, were excised from 4 and 10 day-old seedlings, and used for induction of embryogenic callus. A modified L3 medium, supplemented with 10 M 2,4-dichlorophenoxyacetic acid (2,4-d) for culture initiation, and 5 M 2,4-d for subculturing, was optimal; 90% of 4 day-old Turbo seedlings formed embryogenic callus. Optimal plant regeneration was achieved from callus incubated on a modified MS medium without 2,4-d, but supplemented with 2.22 M 6-benzylaminopurine and 0.27 M naphthaleneacetic acid. Plantlets formed via embryogenesis from all embryogenic Turbo calli initiated from 4 day-old explants, with a mean number of 8 regenerants per explant. Regeneration occured via embryogenesis only. Results obtained using Nandu were within the same range.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid 相似文献
6.
Induction of haploid callus and embryogenesis in in vitro cultured anthers of mulberry (Morus indica) 总被引:1,自引:0,他引:1
Anthers of Morus indica L., with microspores at the uninucleate stage were cultured; and the influence of temperature and kinetin pretreatment on induction of androgenic calluses was examined. The effects of various pretreatments revealed that 24 h cold pretreatment increased the percentage of cultures inducing callus. First microspore division was observed after 16 to 20 days of culture. Th anthers split and developed embryogenic calluses on MB medium supplemented with NAA (0.5 mg l–1 and BA (1.0 mg l–1)) using 8% sucrose. Rhizogenesis was induced on medium supplemented with NAA and BA (each 0.5 mg l–1) with reduced myo-inositol (75 mg l–1). Cytological study of induced roots confirmed the haploid nature of calluses. Different type of embryos were initiated upon transfer of calluses to medium supplemented with NAA, BA (each 0.5 mg l–1), 2,4-d (1.0 mg l–1) and PVP (600 mg l–1). These embryoids further developed roots on removal of 2,4-d from the medium and developed precociously without developing cotyledons and formed elongated shoots.Abbreviations BA
6 benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- FAA
formalin: Acetic acid: Alcohol
- GA3
gibberellic acid
- IBA
indole-3-butyric acid
- MB
modifed Bourgin (Qian et al., 1982)
- NAA
1-naphthalene acetic acid
- PVP
polyvinylpyrrolidone
- RFS-135
rainfed selection 135
- SE
standard error 相似文献
7.
Somatic embryos and embryogenic callus were initiated from immature zygotic embryos of ginseng (Panax ginseng C.A. Meyer). These somatic embryos were multiplied by adventitious (secondary and tertiary) embryogenesis and their growth and development were dependent on growth hormones in the medium. Auxins, 2,4-d, NAA, and IAA at 1.0 mg l-1 were effective in inducing secondary and tertiary somatic embryos, which proliferated directly from the apical or cotyledonary portions of the primary somatic embryos. Single somatic embryos or clusters or embryos developed from the explanted primary embryos. Cytokinin (Kn, BA) inhibited adventitious embryogenesis. Secondary somatic embryos developed to maturation and later regenerated into plantlets in two stage process; firstly elongation of the shoot axes on MS +1.0 mg l-1 Kn, secondly formation of root on 1.0 mg l-1 Kn+1.0 mg-1 GA3 medium.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- IAA
in-doleacetic acid
- Kn
kinetin
- BA
benzylaminopurine
- PSE
primary somatic embryo
- SSE
secondary somatic embryo
- TSE
tertiary somatic embryo 相似文献
8.
Different explants of Miscanthus x ogiformis Honda Giganteus were tested in order to develop an efficient tissue culture system. Shoot apices, leaf and root sections from in vitro-propagated plants, and leaf and immature inflorescence sections from 6-month-old greenhouse-grown plants were used. The explants were cultured on Murashige and Skoog medium supplemented with 4.5, 13.6, 22.6 or 31.7 M 2,4-dichlorophenoxyacetic acid. Three types of callus were formed but only one was embryogenic and regenerated plants. Callus induction and formation of embryogenic callus depended on the type and developmental stage of the explants. Shoot apices formed the highest percentage of embryogenic callus. There was a difference in the formation of embryogenic callus between leaf explants from in vitro-propagated shoots and greenhouse-grown plants. The best results were obtained from newly formed leaves of in vitro-propagated shoots and older leaves of greenhouse-grown plants. Immature inflorescences smaller than 2.5 cm produced a higher percentage of embryogenic callus than larger more mature inflorescences. Embryogenic callus derived from immature inflorescences had the highest regeneration capacity. Differences in 2,4-dichlorophenoxyacetic acid concentrations had no significant effect on callus induction, embryogenic callus formation and plant regeneration.Abbreviations MS
Murashige & Skoog
- 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzyladenine
- NAA
1-naphthaleneacetic acid
- PPFD
photosynthetic photon flux density 相似文献
9.
Archana Giri Paramvir Singh Ahuja P. V. Ajay Kumar 《Plant Cell, Tissue and Organ Culture》1993,32(2):213-218
Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall. Callus was induced on a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid (2,4-d 1 mg l-1) and kinetin (KN 0.5 mg l-1) with coconut water (CW 10% v/v) or naphthalene acetic acid (NAA 5 mg l-1) and benzylaminopurine (BAP 1 mg l-1). Somatic embryos appeared after 2–3 months or 2 subculture passages when 2,4-d or NAA induced source of the callus was transferred to a MS medium containing BAP (1 mg l-1) and NAA (0.1 mg l-1). For successful plantlet formation, the somatic embryos were transferred to a medium containing 1/4 strength MS nutrient with indole-3-butyric acid (IBA 1 mg l-1). Alternatively, the somatic embryos were dipped in a concentrated solution of IBA for 5 min and placed on a hormone free medium. Complete plantlets were formed after 4 weeks and were transferred successfully to soil.CIMAP Publication No. 1020. 相似文献
10.
D. H. Tejavathi M. D. Rajanna R. Sowmya K. Gayathramma 《In vitro cellular & developmental biology. Plant》2007,43(5):423-428
Somatic embryogenesis from cultures of shoot apices, cotyledon and young leaves of in vitro shoots of Agave vera-cruz Mill. was studied. Embryogenic callus was obtained when explants were cultured on Murashige and Skoog’s (MS) medium (1962)
supplemented with L2 vitamins, 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-d) or 5.37 μM ∝-naphthalene acetic acid (NAA). Somatic embryos differentiated from this embryogenic callus upon subculture
to maturation/conversion medium containing cytokinin either alone or with auxin and l-glutamine. The best combination of growth regulators for development of somatic embryos was found to be 5.37 μM naphthalene
acetic acid plus 0.91 μM zeatin and 40 g/l sucrose. The conversion frequency of somatic embryos to plantlets varied from 46–50%.
Rooted plantlets were transferred directly to pots containing a soil, sand, and manure mixture without any hardening phase
with 96–98% survival of the plantlets. Based on the histological observations, the potential origin of the somatic embryo
is discussed. 相似文献
11.
Konstantin V. Kiselev Anna V. Turlenko Yuri N. Zhuravlev 《Plant Cell, Tissue and Organ Culture》2009,99(2):141-149
A somatic embryogenesis protocol for plant regeneration of northern red oak (Quercus rubra) was established from immature cotyledon explants. Embryogenic callus cultures were induced on Murashige and Skoog medium
(MS) containing 3% sucrose, 0.24% Phytagel™, and various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-d) after 4 weeks of culture in darkness. A higher response (66%) of embryogenic callus was induced on 0.45 μM 2,4-d. Higher numbers of globular- (31), heart- (17), torpedo- (12), and cotyledon-stage (8) embryos per explant were obtained
by culturing embryogenic callus on MS with 3% sucrose, 0.24% Phytagel™, and devoid of growth regulators after 8 weeks culture
in darkness. Continuous sub-culturing of embryogenic callus on medium containing 2,4-d yielded only compact callus. Desiccation of embryos for 3 days in darkness at 25 ± 2°C followed by cold storage at 4°C in
darkness for 8 weeks favored embryo germination and development of plantlets. Cotyledon-stage embryos subjected to desiccation
and chilling treatment cultured on MS with 3% sucrose, 0.24 Phytagel™, 0.44 μM 6-benzylaminopurine (BA), and 0.29 μM gibberellic
acid germinated at a higher frequency (61%) than with 0.44 μM BA alone and control cultures. Germinated plantlets developed
a shoot and root, were acclimatized successfully, and maintained in a growth room for plantlet development. 相似文献
12.
L. Semeria B. Ruffoni M. Rabaglio A. Genga A. M. Vaira G. P. Accotto A. Allavena 《Plant Cell, Tissue and Organ Culture》1996,45(1):67-72
Callus cultures were initiated from micropropagated Artemisia absinthium plantlets on MS basal medium supplemented with different concentrations of BA, Kn, NAA, IAA and 2,4-d in combination or singly. Supplementing the medium with low doses of both BA in combination with NAA, and Kn in combination with NAA enhanced the growth rate of callus cultures. However, cultures grew slowly following the second subculture and the majority turned brown and died within the next month. Initiation of root and shoot primordia occured directly from leaf explants cultured on 1.81 M 2,4-d, while adventitious shoot formation from callus was observed occasionally when BA was added to the medium in combination with IAA. Furthermore, medium containing 2.22 M BA and 2.69 M NAA stimulated both callus growth and organogenesis on some callus cultures derived from leaves and stems of young stock material. The best results were obtained with leaf explants. Cytological analysis of root meristems revealed that all regenerants were diploid (2n=18), as expected.Abbreviations MS
Murashige & Skoog's salts and vitamins (1962)
- BA
6-benzyladenine
- NAA
alphanaphthaleneacetic acid
- Kn
Kinetin (6-furfurylaminopurine)
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indoleacetic acid
- FW
fresh weight
- Bi
biomass increase 相似文献
13.
Plant regeneration via somatic embryogenesis in ginger 总被引:5,自引:0,他引:5
A. Kackar S. R. Bhat K. P. S. Chandel S. K. Malik 《Plant Cell, Tissue and Organ Culture》1993,32(3):289-292
Embryogenic callus cultures of ginger were induced from young leaf segments taken from in vitro shoot cultures. Among the four auxins tested in Murashige & Skoog medium, dicamba at 2.7 M was most effective in inducing and maintaining embryogenic cultures. Efficient plant regeneration was achieved when embryogenic cultures were transferred to Murashige & Skoog medium containing 8.9 M benzyladenine. Histological studies revealed various stages of somatic embryogenesis characteristic of the monocot system. The in vitro-raised plants have been established in soil.Abbreviations BA
benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog
- NAA
naphthaleneacetic acid 相似文献
14.
Barbara Stefaniak 《Plant cell reports》1994,13(7):386-389
Summary Friable embryogenic callus and somatic embryos of 4 Gladiolus cultivars were obtained on Murashige and Skoog (MS) medium with various concentration of auxins from the following explants: corm slices, young leaf bases and whole, intact plantlets. Somatic embryos transferred on MS hormone-free medium regenerated into plantlets. All plantlets obtained through embryogenesis did not differ phenotypically from the parental clones. The embryogenic friable callus has been maintained for over 2 years in culture and has retained a very high regeneration capacity.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- KIN
kinetin
- NAA
naphthaleneacetic acid
- MS
Murashige and Skoog Medium (1962)
- E
embryogenic callus
- NE
non-embryogenic callus 相似文献
15.
Fertile regenerated plants were obtained from protoplasts via somatic embryogenesis in Coker 201 (Gossypium hirsutum L.). Protoplasts were isolated from six different explantsleaves, hypocotyls, young roots, embryogenic callus, immature somatic embryos and suspension cultures and cultured in liquid thin layer KM8P medium. Callus-forming percentage of 20–50% was obtained in protoplast cultures from embryogenic callus, immature embryos and suspension cultures, and visible callus formed within 2 months. Callus-forming percentage of 5–20% in protoplast cultures from young roots, hypocotyls and leaves, and visible callus formed in 3 months. NAA 5.371 μM/kinetin 0.929 μM was effective to stimulate protoplast division and callus formation from six explants. Percentage of callus formation in the medium with 2,4-D 0.452 μM/kinetin 0.465 μM was over 40% from suspension cultures and immature embryos, 25% from embryogenic callus and 10% from hypocotyls. Callus from protoplasts developed into plantlets via somatic embryogenesis. Over 100 plantlets were obtained from protoplasts derived from 6 explants. Ten plants have been transferred to the soil, where they all have set seeds. 相似文献
16.
Somatic embryogenesis from pea embryos and shoot apices 总被引:3,自引:0,他引:3
Conditions were defined for plant regeneration via somatic embryogenesis in pea, using explants from immature zygotic embryos or from shoot apices. For the induction of somatic embryos, an auxin (picloram or 2,4-dichlorophenoxyacetic acid) was required. Embryogenic callus originated from embryonic axis tissue of immature embryos and from the axillary-bud region and the plumula of shoot apices. A clear effect of embryo size on somatic embryogenesis was shown. There were differences in frequency of somatic embryogenesis among the five genotypes used in the study. Additions of BA to auxin-containing medium reduced embryo production. Histological examinations confirmed the embryogenic nature of the immature embryo cultures and revealed that somatic embryos originated from the meristematic areas near the callus surface.Abbreviations BA
benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- picloram
4-amino-3,5,6-trichloropicolinic acid 相似文献
17.
Plant regeneration from four genotypes of weeping lovegrass (Eragrostis curvula (Schrad.) Nees), is reported via three developmental pathways: embryogenesis, organogenesis and direct regeneration. Organogenic and embryogenic callus cultures were initiated from young inflorescence segments on Murashige and Skoog's medium supplemented with 2,4-d and BA at different concentrations. The most suitable concentrations of 2,4-d for callus growth and development were 9 and 18 M combined with a BA concentration of 0.044 M. Genotypical differences were observed in the morphogenetic capacity. Direct regeneration was observed under similar culture conditions (culture medium, temperature and photoperiod) but with high light intensity (66 mol m-2 s-1). Young plants were successfully transplanted to pots and grown to maturity in the greenhouse.Abbreviations BA
benzyladenine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog medium
- NAA
naphthaleneacetic acid 相似文献
18.
Charlene Chang Ben A. Moll Kathleen B. Evenson Mark J. Guiltinan 《Plant Cell, Tissue and Organ Culture》1996,45(1):61-66
In vitro plantlet regeneration systems for the seed geranium (Pelargonium x hortorum Bailey) using cotyledon, hypocotyl and root explants were optimized by studying the influence of seedling age, growth regulators and excision orientation on organogenesis. Indole-3-acetic acid combined with zeatin yielded the highest rate of shoot production on cotyledon explants (0.2–2 shoots per explant). More shoots were produced on explants cut from the most basal region of cotyledons from 2 to 4-day-old seedlings than from older seedlings or more distal cut sites. Hypocotyl explants produced the highest number of shoots, up to 40 shoots per explant, on indole-3-acetic acid (2.8–5.6 mM) + zeatin (4.6 mM) or thidiazuron (4.5 mM). Maximum shoot formation (0.3–1.4 shoots per explant) on root explants occurred when they were cultured on medium containing zeatin. Regenerated shoots rooted best on a basal medium containing no growth regulators. There were substantial differences among cultivars in shoot formation from each of the explant systems.Abbreviations BA
6-benzylaminopurine
- 2,4-d
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- NAA
naphthaleneacetic acid
- TDZ
thidiazuron 相似文献
19.
Somatic embryogenesis and plant regeneration in cultured immature inflorescences of Setaria italica 总被引:1,自引:0,他引:1
Young inflorescence explants of Setaria italica in culture showed high capacity for regenerating plantlets through somatic embryogenesis. Embryogenic callus formation was initiated from the explants cultured on Murashige and Skoog's medium with 2 mg/l 2,4-D and 0.2–0.5 mg/l KT or BAP, but it was better for the maintenance of embryogenic growth to subculture the calli on the medium with 2,4-D and KT/BAP and on the medium with 2 mg/l 2iPA and 0.2 mg/l NAA alternately. A number of plantlets were regenerated when embryogenic calli were transferred onto the same basic medium but with 2 mg/l BAP and 0.5 mg/l NAA. Plant regeneration capacity has been maintained in some embryogenic calli during fourteen months of subculture.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- IAA
3-indoleacetic acid
- 2iPA
N6-(2-isopentenyl) adenosine
- BAP
6-benzylaminopurine
- KT
kinetin
- CH
casein hydrolysate 相似文献
20.
Isabel Santos Isabel Guimarães Roberto Salema 《Plant Cell, Tissue and Organ Culture》1994,37(1):83-86
Leaf explants of Nerium oleander L. produced masses of callus when both an auxin and a cytokinin were included in the medium. Leaves cultured on the B5 medium of Gamborg et al. supplemented with 2,4-dichlorophenoxyacetic acid (2,4-d; 9.05 M) plus benzyladenine (BA; 4.4 M) produced callus and profuse rhizogenesis was observed from callus developed from older leaves. On Murashige & Skoog medium (MS) with the same concentration of 2,4-d and BA, explants from young and mature leaves produced callus, but only that from young leaves was embryogenically competent. Globular somatic embryos were obtained when embryogenic cells were cultured on MS medium without growth regulators. Both normal and anomalous development of embryos occurred in either liquid or gelled medium. Plantlets were produced faster when mature embryos were cultured on either solid medium or placed on Sorbarod plugs soaked with this same medium but with 1% sucrose. Plantlets with three nodes were transferred to pots and acclimatized in a growth chamber and afterwards transferred to garden beds. 相似文献