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1.
A short-term regeneration system from leaf-base-derived callus of wheat (Triticum aestivum L.) was developed. Embryogenic callus formation and shoot regeneration were achieved from the first basal segments of 3–4-day-old seedlings. Callus formation frequency as well as plantlet regeneration frequency was dependent on the composition of basal medium and the concentration of 2,4-dichlorophenoxyacetic acid (2,4-D). MS medium with 2,4-D 4.5–9.0 mol l–1 was optimal for the culture of wheat leaf base. Effects of different combinations of plant growth regulators, which were added in either callus induction medium or shoot regeneration medium, were tested. Adding of BAP in callus induction medium shortened the time of shoot emergence but could not improve the producing of embryogenic calli and green plantlets. Optimal ratio of 2,4-D, BAP and NAA gave similar regeneration frequency to control. Existence of cytokinins in regeneration medium had no effect on increasing the regeneration frequency. The regenerants could grow to normal, fertile plants after they were transferred into soil. 相似文献
2.
Yuexia Wang Bridget A. Ruemmele Joel M. Chandlee W. Michael Sullivan Jane E. Knapp Albert P. Kausch 《In vitro cellular & developmental biology. Plant》2002,38(5):460-467
Summary Embryogenic callus induction and plant regeneration systems have long been established for creeping bentgrass (Agrostis palustris Huds.), but little research has been reported on optimal medium for embryogenic callus induction and plant regeneration in
velvet bentgrass (Agrostis canina L.), colonial bentgrass (Agrostis capillaries L.), and annual bluegrass (Poa annua L.). The present study compared 14 callus induction media and eight regeneration media for their efficacies on embryogenic
callus induction and plant regeneration in these four species. The embryogenic callus initiation media contained the Murashige
and Skoog inorganic salts and vitamins supplemented with 2,4-dichlorophenoxyacetic acid or 3,6-dichloro-anisic acid and 6-benzyladenine.
l-Proline or casein hydrolyzate was included in some media to stimulate embryogenic callus formation and plant regeneration.
The frequencies of embryogenic callus formation ranged from 0% to 38% and exhibited medium differences within each of the
four species. Callus induction media, plant regeneration media, and genotypes affected plant regeneration rates, which varied
between 0% and 100%. The embryogenic callus induced on Murashige and Skoog medium supplemented with 500 mgl−1 casein hydrolyzate, 6.63 mg l−1 (30 μM) 3,6-dichloro-anisic acid and 0.5–2.0 mg l−1 (2–9 μM) 6-benzyladenine had much higher regeneration rates than those formed on other callus induction media. Embryogenic callus
of annual bluegrass had higher regeneration rates than those of bentgrass species. MSA2D, a media containing 2 mgl−1 (8 μM) 2,4-dichlorophenoxyacetic acid, 100 mgl−1
myo-inositol, and 150 mgl−1 asparagine, was effective in promoting embryogenic callus formation in creeping bentgrass but not in colonial and velvet
bentgrasses and annual bluegrass. 相似文献
3.
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
4.
Optimization of the conditions for an efficient induction of somatic embryogenic calli and regeneration of plants from mature seeds of japonica rice cultivars was attempted. The number, color, size, shape, and appearance time of the induced embryogenic calli varied among the rice cultivars depending on the type of basal medium (LS, MS, N6). Presence of adequate amount of sucrose in the medium was an absolute requirement for embryogenic callus formation and shoot induction. Induction of the embryogenic calli, whose overall rates ranged from 30 to 56%, was most efficient in N6 medium supplemented with 3.0 mg l–1 of 2,4-D and 30 g l–1 of sucrose. Agar concentration in the regeneration medium was also critical for the shoot induction. Kinetin was found to be more effective for shoot regeneration compared with BA, while the highest shoot regeneration frequencies were observed when either cytokinin was combined with high concentration (2.0 mg l–1) of NAA. The optimal concentration of kinetin for the highest shoot regeneration frequency (6777%) was different among the cultivars tested. The embryogenic calli-derived shoots rooted on a plant growth regulator-free MS medium were successfully established in soil, producing fertile seeds. 相似文献
5.
Joseph C. V. Vu Randall P. Niedz George Yelenosky 《Plant Cell, Tissue and Organ Culture》1993,35(1):75-80
Anthers of niger (Guizotia abyssinica. Cass) were inoculated onto five different media differing mainly in their inorganic and organic constituents and plant growth regulators to study their influence on callus induction (embryogenic/non-embryogenic) and plant regeneration. LS medium supplemented with 2 mg 1-1 2,4-d, and 0.3 mg 1-1 KN favoured the production of EC, whereas 2 mg 1-1 BAP and 0.5 mg 1-1 KN promoted the NEC from anthers. Different types of embryos were initiated upon transfer of EC to Chaleff's R-2 medium containing 2 mg 1-1 NAA and 0.3 mg 1-1 KN and/or 5 mg 1-1 ABA. NEC when transferred onto the medium supplemented with 1 mg 1-1 BAP and 0.1 mg 1-1 NAA produced on an average 8–12 shoots/callus mass. Embryoids developed from the EC and shoots differentiated from NEC when cultured onto the Chaleff's R-2 and MS media respectively lacking growth regulators, they transformed into whole plantlets. The plantlets thus obtained were successfully hardened and grown to maturity for analysis of various plant characters.Abbreviations EC
embryogenic callus
- NEC
non-embryogenic callus
- 2,4-d
2,4-dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- ABA
abscisic acid
- BAP
6-benzylaminopurine
- KN
kinetin
- MS
Murashige and Skoog's medium
- LS
Linsmaier and Skoog's medium 相似文献
6.
Ivan Rodrigo Artunduaga Charles M. Taliaferro Becky L. Johnson 《Plant Cell, Tissue and Organ Culture》1988,12(1):13-19
Objectives of this research were to test the effects of plant genotypes and auxin 2,4-D (2,4-dichlorophenoxyacetic acid) medium concentrations on embryogenic (E) callus production of two grass species. Two Old World bluestem,Bothriochloa ischaemum, accessions (A-8793 and A-8911c) and three bermudagrass,Cynodon dactylon (L.) Pers., accessions (A-10978b, A12164, and Brazos) supplied the explant material. Immature inflorescences 9 mm in length were placed on modified Murashige-Skoog (MS) agar medium containing 0, 1, 3, or 5 mg L-1 of 2,4-D. Explants of all genotypes produced callus by the end of a 4-week dark incubation period at 25°C. When subcultured onto fresh media and maintained at 25°C with a 16 hr photoperiod, calli became embryogenic within 8 weeks of inoculation. Three mg L-1 of 2,4-D in the media maximized E callus production in both bluestem genotypes and in A-10978b and A-12164 bermudagrass genotypes. Maximum E callus production from Brazos bermudagrass resulted from the 1 mg L-1 treatment. Somatic embryos developed after subculture under light. Embryos showed scutellum-like structures and coleoptile-coleorhiza bipolar organization. Plantlets were regenerated from all genotypes except Brazos, whose embryoids failed to germinate. All callus from Brazos eventually senesced. Light and scanning electron microscopy confirmed regeneration through somatic embryogenesis. 相似文献
7.
Callus was induced from hypocotyl and primary leaf explants of cumin (Cuminum cyminum L.) seedlings on a medium with 4 M 2,4-D alone or plus 2 or 4 M kinetin. An embryogenic callus developed within 2 weeks after transferring the callus to medium lacking plant growth regulators (PGR). The presence of kinetin in the callus induction medium with 2,4-D enhanced both the callus proliferation and the subsequent differentiation of the embryoids on the PGR-free medium. Plumules with or without simultaneously developed roots were observed 3–4 weeks after subculturing the embryogenic callus on medium containing 0.5 or 1.0 M kinetin. Subsequently, they were transferred onto half-strength medium supplemented with 1 M indole-3-butyric acid (IBA) and 2% polyethylene glycol (PEG, 6000) for root induction and/or proliferation, and in vitro hardening of the regenerated plants. The survival rate ex vitro was 70%. No plants developed from the embryogenic callus continuously incubated on medium lacking kinetin. We concluded that kinetin is crucial for plant regeneration from the induced embryoids of cumin. 相似文献
8.
Q. C. Liu H. Zhai Y. Wang D. P. Zhang 《In vitro cellular & developmental biology. Plant》2001,37(5):564-567
Summary Using 15 Chinese and Japanese cultivars of sweetpotato, Ipomoea batatas (L.) Lam., we succeeded in developing an efficient plant regeneration system from embryogenic suspension cultures. The embryogenic
callus derived from shoot apices of the 15 cultivars was used to initiate embryogenic suspension cultures in Murashige and
Skoog (MS) medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Rapidly proliferating and well-dispersed embryogenic suspension cultures were established.
Cell aggregates 0.7–1.1 mm in size from embryogenic suspension cultures were transferred to solid MS medium supplemented with
9.05 μM of 2,4-D and formed embryogenic callus with somatic embryos. The embryogenic callus with somatic embryos was further transferred
to MS medium supplemented with 3.78 μM of abscisic acid, resulting in the germination of somatic embryos. Within 20 wk after the initiation, the frequencies of
cell aggregates forming plantlets reached approximately 100% for the 15 tested cultivars. These plantlets, when transferred
to soil, showed 100% survival. No morphological variations were observed. 相似文献
9.
K. Haliloglu 《Biologia Plantarum》2006,50(3):326-330
Efficient plant regeneration system from leaf base segments of wheat (Triticum aestivum L.) was developed. The factors affecting the callus formation and regeneration capacity of leaf segments of two genotypes;
Bobwhite and Pavon 76, were investigated. The highest number of somatic embryos (SE) was obtained on Murashige and Skoog medium
supplemented with 2 mg dm−3 2,4-dichlorophenoxyacetic acid + 1 mg dm−3 naphthalenacetic acid (14.7 SE per segment). Highest frequency of embryogenic callus (96 %) and somatic embryo formation
(24.3 SE per segment) were achieved in the first segments. The highest plantlet regeneration was obtained after transfer of
embryogenic calli to regeneration medium supplemented with 1 mg dm−3 kinetin (6.3 plantlets per segment). 相似文献
10.
Summary The plant regeneration ability of callus obtained from zygotic embryos of the monocot Alstroemeria spp. was studied. The best explants for somatic embryogenesis were immature zygotic embryos in half-ovules when the endosperm was still soft and white. For 2 genotypes embryogenic callus was induced on callus induction medium with a success rate of 54%. The best callus induction period was 10 weeks. The morphology of embryogenic callus was nodular. Somatic embryos were formed after transfer of the callus to regeneration medium. These somatic embryos revealed later on the typical features of zygotic Alstroemeria embryos. The total duration of the plant regeneration protocol, from inoculation till rooted plantlets ready for transfer to the greenhouse, was 28 weeks.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962)
- NAA
-naphthaleneacetic acid 相似文献
11.
High frequency somatic embryogenesis and plant regeneration from zygotic embryo-derived callus cultures of three Allium species 总被引:1,自引:0,他引:1
P. van der Valk O. E. Scholten F. Verstappen R. C. Jansen J. J. M. Dons 《Plant Cell, Tissue and Organ Culture》1992,30(3):181-191
The plant regeneration ability of zygotic embryo-derived callus cultures was studied for 12 A. cepa varieties and accessions, two A. fistulosum varieties, one A. fistulosum x A. cepa interspecific hybrid and two A. porrum varieties. Compact embryogenic callus was induced on Murashige and Skoog (MS) medium supplemented with 2,4-dichlorophenoxyacetic acid. The embryogenic calluses of all three Allium species were similar in appearance. For all accessions tested plants could be regenerated at a high frequency from this compact callus through somatic embryogenesis, when using kinetin supplemented MS medium (regeneration medium). Addition of abscisic acid to the regeneration medium stimulated the formation of both somatic embryos and shoots for a number of varieties. Concerning shoot regeneration from callus cultures, significant differences existed between genotypes of all accessions except one.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- VDH
Van Der Have Seed company 相似文献
12.
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
13.
J. B. Teixeira M. R. Söndahl T. Nakamura E. G. Kirby 《Plant Cell, Tissue and Organ Culture》1995,40(2):105-111
Primary globular callus from immature zygotic embryos and friable embryogenic tissue derived from mature zygotic embryos were used to establish suspension cultures. Callus cultures were established either on modified Y3 or MS medium containing 475–500 M 2,4-D or 250 M picloram and 0.3% (w/v) activated charcoal. Suspension cultures of both cell lines were established in modified Y3 medium containing 10 M 2,4-D. The establishment of cell suspensions from friable embryogenic tissue took only 2 months, in contrast with suspensions from primary globular callus which took 3–5 months to establish. Embryo differentiation was observed only in cell suspensions derived from the friable embryogenic tissue after plating aliquots on regeneration medium. Germinated embryos were recovered and plantlets were successfully established under greenhouse conditions.Abbreviations CET
compact embryogenic tissue
- FET
friable embryogenic tissue
- CIM
callus induction medium
- PGC
primary globular callus
- 2,3-D
2,4-dichlorphenoxyacetic acid Y3-Eeuwens' medium
- MS
Murashige & Skoog medium
- PVP-40
polyvinylpyrrolidone
- KM
Kao & Michayluk vitamins
- ABA
abscisic acid 相似文献
14.
Control of direct and indirect somatic embryogenesis by exogenous growth regulators in immature zygotic embryo cultures of rose 总被引:3,自引:0,他引:3
Immature zygotic embryos of rose (Rosa hybrida L.; cv. Sumpath) did not form somatic embryos or embryogenic calluses when cultured on half-strength Murashige and Skoog's medium supplemented with various con-centrations of 2,4-dichlorophenoxyacetic acid (2,4-D) as the sole growth regulator. However, the zygotic embryos produced somatic embryos without an intervening callus phase at a frequency of 27.3% on medium with 4.44 M 6-benzyladenine (BA) alone. Immature zygotic embryos formed embryogenic calluses at a frequency of 25% on medium with a combination of 1.36 M 2,4-D and 4.44 M BA. Upon transfer to medium without growth regulators, embryogenic calluses produced numerous somatic embryos that subsequently developed into plantlets. Somatic embryos were induced directly from immature zygotic embryos, or indirectly via an intervening callus phase, by manipulating the exogenous growth regulators. Plantlets were successfully transplanted to potting soil and grown to maturity in a greenhouse. 相似文献
15.
An improved procedure for the induction, proliferation and regeneration of embryogenic callus from coffee leaf explants has been developed. The optimal culture conditions for callus induction and somatic embryogenesis yielded so-called high frequency embryogenic callus ofCoffea canephora P. ex Fr., Arabusta and Congusta, more rapidly and abundantly than other published procedures.Coffea arabica L. genotypes, however, were less responsive to the procedure. The highest multiplication rate of embryogenic callus in liquid culture, which avoided the differentiation of embryos, was obtained by culture at an inoculum density of 10 g callus 1-1 in a modified MS medium containing 4.5 M 2,4-dichlorophenoxyacetic acid, under 3 mol m-2 s-1 illumination, and subcultured every 7–10 days. The best long-term maintenance of embryogenic potential was obtained by culture of aggregates (250–1000 m in diameter) at an inoculum density of 5 g 1-1, with medium renewed every 3–4 weeks. Under these conditions, embryogenic potential ofC. canephora callus was maintained for over 2 years. Analysis of nutrients absorbed by the callus cultures demonstrated that half strength MS macro- and micro-salts were not depleted during at least 3 weeks of sustained culture. The highest regeneration of embryogenic callus required the omission of 2,4-D and a reduced culture density of 1 g 1-1. Under these conditions of culture, 1 g ofC. canephora or Arabusta callus produced 1.2 and 0.9×105 somatic embryos, respectively, after 8–10 weeks in liquid regeneration medium. This was an overall reduction of 4–6 months from explant to regenerant, when compared with other procedures.Abbreviations BA
N6-benzyladenine
- HFSE
high frequency somatic embryogenesis
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- rpm
rotations per minute
- LFSE
low frequency somatic embryogenesis
- MS
Murashige & Skoog medium
- PPF
photosynthetic photon flux
- 2,4-D
2,4-dichlorophenoxyacetic acid
- 2-iP
2-isopentenyladenine 相似文献
16.
High frequency plant regeneration via somatic embryogenesis has been induced from in vitro shoot-base cultures of seedlings of garden leek (Allium porrum L.). Four main steps are involved in the procedure using BDS medium: - shoot multiplication with 17.6 mM benzyladenine; - induction of nodular callus from the in vitro shoot base with 9 mM 2,4-dichlorophenoxyacetic acid; - initiation of embryogenic callus from nodular callus with 9 mM 2,4-dichlorophenoxyacetic acid +7.6 mM abscisic acid; - plant regeneration from embryogenic callus with 9.8 mM N6-(2-isopentenyl)adenine. The presence of 2,4-dichlorophenoxyacetic acid in the medium and light conditions were shown to be essential for nodular callus induction and somatic embryogenesis. Abscisic acid was not a prerequiste for somatic embryogenesis, but it significantly increased the frequency. 相似文献
17.
High-frequency plant regeneration through callus initiation from mature embryos of maize (<Emphasis Type="Italic">Zea Mays</Emphasis> L.) 总被引:10,自引:0,他引:10
An efficient maize regeneration system was developed using mature embryos. Embryos were removed from surface-sterilized mature seeds and sliced into halves. They were used as explants to initiate callus on induction medium supplemented with 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D). The induction frequency of primary calli was over 90% for all inbred lines tested. The primary calli were then transferred onto subculture medium supplemented with 2.0 mg l–1 2,4-D. Following two biweekly subcultures, embryogenic calli were formed. Inclusion of a low concentration (0.2 mg l–1) of 6-benzylaminopurine (BA) in the subculture medium significantly promoted the formation of embryogenic callus. The addition of silver nitrate (10 mg l–1) also supported an increased frequency of embryogenesis. The embryogenic callus readily formed plantlets on regeneration medium supplemented with 0.5 mg l–1 BA. The regenerated plantlets were transferred to half-strength Murashige and Skoog medium supplemented with 0.6 mg l–1 indole-3-butyric acid to develop healthy roots. The regenerated plantlets were successful on transfer to soil and set seed. Using this system, plantlets were regenerated from seven elite maize inbred lines. The frequency of forming green shoots ranged from 19.8% to 32.4%. This efficient regeneration system provides a solid basis for genetic transformation of maize.Abbreviations BA 6-Benzylaminopurine - 2,4-D 2,4-Dichlorophenoxyacetic acid - IBA Indole-3-butyric acid - KT KinetinCommunicated by M.C. Jordan 相似文献
18.
Shyamkumar Barampuram Byung Yeoup Chung Seung Sik Lee Byung Chull An Eun Mi Lee Jae-Young Cho 《In vitro cellular & developmental biology. Plant》2009,45(2):155-161
The present study demonstrates the establishment of embryogenic tissue from seeds and (seedling-derived hypocotyls) shoot
base explants derived from seedlings of Eremochloa ophiuroides. The highest percentage of callus induction obtained from seed and young shoot base explants was 52.0% and 66.6% on Murashige
and Skoog (MS) basal media supplemented with 9.0 μM and 18.1 μM 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. The
type of callus obtained from both types of explants was off-white to yellow in color and non-friable and shiny in texture.
Excised callus from the explants was subcultured onto fresh media of the same recipe for further proliferation. After 10–12 d
of subculture, a yellow, globular, friable embryogenic callus was obtained from the initial callus. The highest percentage
of embryogenic calli obtained at 40.0% was observed on media containing 2.2 μM 2,4-D. The highest regeneration rate of 46.6%
was observed on MS media supplemented with 0.4 μM 2,4-D and 2.2 μM benzylaminopurine (BA). Regenerated shoots were rooted
in MS basal medium. Plants with well-developed roots were transferred to pots containing a soil mix and acclimatized in greenhouse
conditions. Four weeks post-transfer, acclimatized plants showed 100% survival and remained healthy and green. This is the
first report of a successful method for induction of somatic embryogenesis with subsequent plant regeneration in centipede
grass and demonstrates the establishment of embryogenic callus and efficient plant regeneration with potential application
in the development of genetic transformation systems for centipede grass. 相似文献
19.
A. S. T. Immonen 《Plant Cell, Tissue and Organ Culture》1996,44(1):45-52
Basal media and plant growth regulators were tested for the promotion of somatic embryogenesis from immature wheat-rye hybrid embryos. Influence of growth regulators and chilling on plant regeneration were tested on two media. A medium containing four amino acids-glutamine, arginine, glycine and aspartic acid-as the nitrogen source, promoted the production of, on average, twice as much embryogenic callus as the other media, and somatic embryos developed well. The growth regulator dicamba was significantly better than 2,4-dichlorophenoxyacetic acid in promoting somatic embryogenesis and subsequent plant regeneration. Germination of somatic embryos on both regeneration media was enhanced by cold treatment. Supplementing 190-2 plant regeneration medium with a combination of -naphthaleneacetic acid + benzyladenine, indole-3-acetic acid + kinetin or indole-3-acetic acid + zeatin resulted in equally high germination rates.Abbreviations 190-2
Plant regeneration medium of Chuang & Jia
- 2,4-d
2,4d Dichlorophenoxyacetic acid
- Dicamba
3,6-Dichloro-o-anisic acid
- AA
Amino acid medium of Müller & Grafe
- IAA
Indole-3-acetic acid
- BA
Benzyladenine
- NAA
-Naphthaleneacetic acid 相似文献