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1.
Callus was induced on the wounded immature seeds and mature zygotic embryos of Dysosma pleiantha (Hance) Woodson (Berberidaceae) on a medium based on Murashige and Skoog's (1962) formula supplemented with 1 mg/l 2,4-dichlorophenoxy-acetic acid (2,4-D). Spontaneous embryoid formation occurred on the media containing low concentrations of 2,4-D (0.1–0.5 mg/l). These embryoids germinated in either MS or B5 medium containing 1 mg/l N6-benzyladenine and 1 mg/l gibberellic acid. The regenerated plantlets were successfully transferred to soil.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
N6-benzyladenine
- GA3
gibberellic acid
- MS medium
Murashige and Skoog's (1962) medium 相似文献
2.
Plantlet regeneration through indirect somatic embryogenesis was attempted from rhizome derived callus of Cymbopogon winterianus Jowitt (cv. Jorlab2). Optimum callus was induced on Murashige and Skoog (MS) basal medium supplemented with 4 mg dm−3 2,4-dichlorophenoxyacetic acid (2,4-D). Initially the callus was friable, shiny white and watery in nature. After subculturing
on MS medium containing 2,4-D and kinetin (Kn), callus was transferred onto the MS medium supplemented with 2,4 -D, Kn and
coconut water to induce somatic embryogenesis. Optimum somatic embryogenesis (78.33 %) was achieved on MS medium containing
3.0 mg dm−3 2,4-D and 0.5 mg dm−3 Kn. High frequency (65 %) plantlet conversion from embryos was achieved in MS medium supplemented with 2 mg dm−3 N6-benzyladenine (BA), 0.5 mg dm−3 Kn, 0.2 mg dm−3 calcium pantothenate and 0.2 mg dm−3 biotin. 相似文献
3.
Plant regeneration from protoplast culture of Crocus cancellatus was investigated using regenerable embryogenic calli obtained from shoot meristem culture on LS (Linsmaier and Skoog, 1965)
medium containing 4 mg l−1 kinetin and 1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Protoplasts were isolated directly from embryogenic calli. The best protoplast growth
was found on those embedded in Ca-alginate beads and cultured with nurse cells in MS (Murashige and Skoog, 1962) medium supplemented
with 2 mg l−1 kinetin, 1 mg l−1 2,4-D and 100 mg l−1 ascorbic acid at 25 °C in darkness. After 4–5 weeks of culture, microcalli appeared on the surface of the Ca-alginate beads,
but the protoplasts without immobilization in Ca-alginate beads showed very low cell division. Growth of the microcalli in
the medium with nurse cells was much better than in the medium without nurse cells. Transferring beads onto half strength
MS medium supplemented with 0.2 mg l−1 kinetin and 0.1 mg l−1 2,4-D, increased the growth of embryogenic calli. Somatic embryo development was observed either on half strength MS medium
growth regulator free or with 1 mg l−1 abscisic acid. Matured embryos germinated on half strength MS medium containing 25 mg l−1 of gibberelic acid. Plantlet formation was obtained on half strength MS medium containing 1 mg l−1 6-benzyladenine and 1 mg l−1 α-naphthaleneacetic acid at 20 °C in a 16/8 h light/dark cycle.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
4.
Two plant regeneration methods applicable to Leucaenaleucocephala were developed. In the first method, involvingorganogenesis via callus formation, cotyledon, hypocotyl and root segments wereinitiated on MS medium containing different concentrations ofN6-benzyladenine (BA), 2,4-dichlorophenoxyacetic acid (2,4-D), andnaphthaleneacetic acid (NAA). Both compact (type I) and friable (type II) calliwere obtained from the cotyledon and hypocotyl explants treated with differentconcentrations of the growth regulators. Shoots were generated only from thefriable calli formed from the cotyledon explants. The calli formed from thehypocotyl explants did not generate shoots and the root explants died withoutforming callus. Cotyledon explants from 3–4 day old seedlings showedmaximum callus induction compared to those from older seedlings. In a secondmethod involving direct organogenesis, excised cotyledons were cultured on 1/2MS medium containing 10–35 mg l–1N6-benzyladenine (BA) for 7–14 days. Transfer of thecotyledonsto regeneration medium containing low BA resulted in callus formation andsubsequent shoot regeneration from the base of the excised cotyledon explants,with up to 100% frequency. Regenerated shoots rooted best on a basal mediumcontaining no growth regulators. 相似文献
5.
Leaf mesophyll protoplasts ofDianthus superbus were cultured at a density of 5 × 104 protoplasts/ml and divided at about 18% plating efficiency in MS liquid medium supplemented with 0.5 mg/L BAP, 2.0 mg/L NAA and 9% mannitol after 2 weeks. Protocolonies formed after 3 to 4 weeks of culture in the dark at 27°C. These colonies were transferred to continuous illumination (21.5 E m–2 sec–1) for 2 weeks where most of the colonies divided to form microcalli, about 2 mm in diameter. Subsequently, green microcalli were transferred to MS solidified medium with 2.0 mg/L 2,4-D that induced shoot-forming calli after 4 weeks. These calli were transferred onto N6-2 medium containing 0.1 mg/L 2,4-D, 0.1 mg/L NAA, 2.0 mg/L kinetin and 2.0 g/L casein hydrolysate and were cultured under light. After 5 weeks the calli gave rise to multiple shoots (10 to 15 per callus). Upon transfer to MS medium containing 2.0 mg/L NAA, individual shoots were rooted in 4 weeks. The regenerants were successfully transplanted into potting soil.Abbreviations MS
Murashige and Skoog
- BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthaleneacetic acid
- N6
Chu basal salt mixture
- MES
2-N-morpholinoethanesulfonic acid 相似文献
6.
The study was carried out to establish in vitro culture conditions for plant regeneration of tef, Eragrostis tef (Zucc.) Trotter. Mature seeds of two Ethiopian varieties, DZ-01-354 and DZ-01-196, were used to initiate callus cultures
on Murashige and Skoog (MS) medium with different auxins. Four- and 8-week-old calli induced on a medium with 2.0 mg/l 2,4-dichlorophenoxyacetic
acid (2,4-D) were subcultured onto various media to induce somatic embryogenesis. Compact, nodulated, embryogenic callus was
observed after transfer onto MS-callus proliferating (CP) medium. Embryogenic tissue appeared on soft and amorphous callus
and developed into somatic embryos during a subsequent subculture to MS embryo-promoting (EP) media. Various growth regulator
combinations were tested in CP and EP media to obtain a high efficiency of somatic embryo formation. The highest frequency
of calli forming somatic embryos (56.1–68.3%) was observed when CP media with 2.0 or 4.0 mg/l 2,3,5-triiodobenzoic acid were
employed and then cultures were transferred to EP media with 0.5 mg/l 2,4-D and 0.5 mg/l kinetin followed by 0.5 mg/l indole-3-acetic
acid and 0.5 mg/l N6-benzyladenine. Plant development from somatic embryos was obtained on MS medium supplemented with 1.0 mg/l gibberellic acid.
On average, 71.2% of calli displaying somatic embryos converted into plants. Regenerated plants were successfully transferred
to soil. Neither chlorophyll-deficient plants nor morphological variants were found among regenerants. All regenerated plants
were fertile.
Received: 9 May 1997 / Revision received: 25 September 1997 / Accepted: 3 January 1998 相似文献
7.
Leaves of Solanum virginianum plants were used for protoplast isolation. To support cell wall formation and cell division, protoplasts were cultured in
thin alginate layers floated in liquid medium. When protoplasts were plated at a density of 1.0 × 106/ml in Kao and Michyaluk (KMp8) medium supplemented with 0.5 mg/l zeatin, 1.0 mg/l 2,4-dichlorophenoxyacetic acid, and 1.0 mg/l
α-naphthaleneacetic acid, 42.3% of the dividing cells developed microcalli in 3–4 weeks. Shoot formation via organogenesis
of protoplast-derived calli was achieved for 28% of calli transferred to solidified KMp8 medium supplemented with 2.0 g/l
zeatin and 0.1 mg/l 3-indol acetic acid in about 2 weeks. Further shoot development was observed in Murashige and Skoog (MS)
medium without growth regulators and roots were induced after transfer to MS medium containing 1.0 mg/l 3-indol butyric acid.
Regenerated plants have normal morphology. 相似文献
8.
Anca-Livia Butiuc-Keul Laurian Vlase Cornelia Cr?ciuna? 《In vitro cellular & developmental biology. Plant》2012,48(2):249-258
In vitro tissue culture protocols were tested for propagation of Echinacea purpurea, Echinacea pallida and Echinacea angustifolia in order to obtain biomass for the production of cichoric acid, which is the major active compound in the Echinacea extracts. The in vitro culture process was initiated by seed germination on half-strength Murashige and Skoog (MS) medium. Multiplication was achieved
on MS medium supplemented with naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), 2-iso-pentenyladenine (2iP), and
N6-benzyladenine (BA) in different concentrations. Shoot explants produced the highest number of shootlets on MS medium, which
was supplemented with 0.1 mg/l 2iP and 0.1 mg/l IBA. RAPD markers revealed genetic polymorphism in some instances between
in vitro generated plantlets such as for E. purpurea plantlets analyzed with the OPO-8 primer. RAPD markers generated with the primer 4A-29 revealed low levels of genetic variation
between in vitro plantlets for all three species of Echinacea, while remaining RAPD markers revealed no variation. Content of cichoric acid in leaves, shoots, and callus was analyzed
by high-performance liquid chromatography/MS and was identified in all studied samples, independent of species or tissue type.
Highest levels (0.39–0.73 mg/g dw) were observed in shoots and leaves. 相似文献
9.
Boon Chin Tan Chiew Foan Chin Peter Alderson 《Plant Cell, Tissue and Organ Culture》2011,105(3):457-463
An indirect in vitro plant regeneration protocol for Vanilla planifolia has been established. Juvenile leaf and nodal segments from V. planifolia were used as explants to initiate callus. Nodal explants showed better callus initiation than juvenile leaf explants, with
35.0% of explants forming callus when cultured on Murashige and Skoog (MS) basal medium supplemented with 2.0 mg/l 1-naphthylacetic
acid (NAA) and 1.0 mg/l 6-benzyladenine (BA). Almost 10.0% of juvenile leaf explants were induced to form callus on the MS
basal medium containing 2.0 mg/l NAA and 2.0 mg/l BA, whereas no callus formed in the presence of any concentrations of 2,4-dichlorophenoxyacetic
acid (2,4-D) and BA. After 8 weeks, callus generated was transferred to MS basal medium containing 1.0 mg/l BA and 0.5 mg/l
NAA. A mean number of 4.2 shoots per callus was produced on this medium, with a mean length of 3.8 cm after 8 weeks of culture.
Roots formed on 88.3% of plantlets when they were cultured on MS medium supplemented with 1.0 mg/l NAA, with a mean length
of 4.4 cm after 4 weeks of culture. Of the rooted plantlets, 90.0% survived acclimatisation and were making new growth after
4 weeks. 相似文献
10.
Somatic embryogenesis was achieved in callus cultures derived from immature cotyledonary explants ofHardwickia binata Roxb., a multipurpose leguminous tree, on semisolid modified Murashige and Skoog's (mMS) medium containing 2900 mg/l potassium nitrate (KNO3) supplemented with 4.64 µM kinetin (Kn) and 5.37µM a-naphthaleneacetic acid (NAA). Somatic embryos proliferated rapidly after transfer to MS basal medium supplemented with 2052.6 µM L-glutamine and 0.084 µM gibberellic acid (GA3). Maturation of somatic embryos was achieved on half-strength MS basal medium supplemented with 1.23 µM IBA and 2% (w/v) sucrose. Histological studies confirmed different developmental stages of somatic embryogenesis inHardwickia binata.
Abbreviations BA
N6-benzyladenine
- Kn
kinetin
- NAA
a-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- IBA
indole-3-butyric acid
- GA3
gibberellic acid
- MS
Murashige and Skoog (1962) medium
- mMS
modified Murashige and Skoog (1962) medium 相似文献
11.
Summary Callus induction was observed from hypocotyl, root, and cotyledonary leaf segments, grown on Murashige and Skoog (MS) medium
supplemented with various concentrations and combinations of 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin (KN). Maximum
callusing (100%) was obtained from root and cotyledonary leaf segments grown on MS medium supplemented with a combination
of 2 mg l−1 (9.1 μM) 2,4-D and 0.2 mg l−1 (0.9 μM) KN. The calluses, when subcultured in the same medium, showed profuse callusing. However, these calluses remained recalcitrant
to regenerate regardless of the quality and combinations of plant growth regulators in the nutrient pool. When hypocotyl segments
were used as explants, callus induction was noticed in 91% of cultures which showed shoot regeneration on MS medium supplemented
with 2 mg l−1 2,4-D and 0.2 mg l−1 KN. These shoots were transferred to fresh medium containing various concentrations and combinations of 6-benzyladenine (BA)
and N6-(2-isopentenyl)adenosine (2-iP). Maximum shoot multiplication was observed after 60 d of the second subculture on MS medium
containing 2 mg l−1 (8.9 μM) BA. These shoots were rooted best (87%) on MS medium containing 2 mg l−1 (9.9 μM) indole-3-butyric acid (IBA). The plantlets were transferred to the field after acclimatization and showed 60% survival. 相似文献
12.
Summary Young leaf segments from plants growing both in vivo and in vitro were cultured on Murashige and Skoog (MS) medium supplemented with auxins [naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic
acid (2,4-D)], cytokinins [kinetin (KN) and N6-benzyladenine (BA)] and coconut liquid endosperm (CW). The explants from mature leaves did not show any growth and turned
necrotic, while those obtained from juvenile leaves growing in vitro developed protocorm-like bodies (PLBs) at their cut surfaces within 4–8 wk depending on the growth medium. An optimum of
18 PLBs developed from leaf explants on medium supplemented with 2.0 mg l−1 (8.87 μM) BA. Upon subculture in basal MS medium, the PLBs differentiated into plantlets within 6–8 wk. The resulting plantlets were
successfully transferred to vermiculite initially and subsequently to potting mixture; 84% of the plantlets survived after
3 mo. of transplantation. 相似文献
13.
Summary Protoplasts were isolated from immature cotyledons of Vigna sinensis and cultured in a modified MS Liquid medium containing 0. 2 mg/l 2, 4-dichlorophenoxyacetic acid (2, 4-D), 1 mg/l naphthaleneacetic acid (NAA) and 0. 5 mg/l 6-benzylaminopurine (BAP) in the dark at a density of 1 × 105/ml. The protoplasts began to divide in 3–5 days. Sustained cell division resulted in formation of cell clusters and small calli, with the cell division frequency and plating efficiency of cell colonies reaching 27. 7% and 1. 7% respectively. When calli of 2 mm in size were transferred onto MSB medium (MS salts and B5 vitamins) containing 500mg/l NaCl, 500 mg/ 1 casein hydrolysate (CH), 2 mg/l 2,4-D and 0. 5 mg/l BAP for further growth, approximately 5% of the calli developed embryogenically. The embryogenic calli were selected and subcultured on the same composition of MSB medium and were able to maintain somatic embryogenesis capacity in subculture for a long time. When the calli were moved to MSB medium with 0. 1 mg/l indole-3-acetic acid (IAA), 0. 5mg/l kinetin(KT), 3–5% mannitol and 2% sucrose in the light, many somatic embryos formed from the calli. Only part of the embryoids developed further to the cotyledonary stage, and the others died at the globular, heart-shaped or torpedo stages. Finally, some cotyledonary embryoids germinated and developed into plants or shoots. The shoots were readily rooted on 1/2 strength MS medium with 0. 1–0.3 mg/l indole-3-butyric acid (IBA). The plants grew well in soil and were fertile.Abbreviations 2, 4-D
dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- KT
kinetin
- IBA
indole-3-butyric acid
- CH
casein hydrolysate
- CM
coconut milk
- ZT
zeatin 相似文献
14.
A rapid protocol for somatic embryogenesis from immature leaflets of groundnut (Arachis hypogaea L.)
P. Venkatachalam P. B. Kavi Kishor N. Geetha M. Thangavelu N. Jayabalan 《In vitro cellular & developmental biology. Plant》1999,35(5):409-412
Summary Plant regeneration via somatic embryogenesis was developed in two groundnut varieties. Somatic embryogenesis was induced from
immature leaflets on MS medium with different concentrations of the auxins 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic
acid (NAA) in combination with 0.5 mg/l of the cytokinin BA. The highest frequency of somatic embryo formation occurred on
MS medium fortified with 20 mg 2,4-D per l. Of the two auxins tested individually 2,4-D was more effective for induction of
embryogenesis as well as production of embryos. Embryo development and maturation was achieved on MS medium supplemented with
N6-benzyladenine (BA) (0.5–2.0 mg/l) and 2,4-D (0.5 mg/l). Plant conversion frequency from somatic embryos was highest in presence
of 2.0 mg BA per l and 0.5 mg NAA per l. The frequency of embryogenesis and plant regeneration was higher in the VRI-2 cultivar
than in the other cultivar tested. Regenerated plants were transferred to soil, grown to maturity, and produced viable seeds. 相似文献
15.
Nasser J. Y. Sholi Anjana Chaurasia Anuradha Agrawal Neera Bhalla Sarin 《Plant Cell, Tissue and Organ Culture》2009,99(2):133-140
Creamy friable calli were induced from meristems (scalps) of proliferating shoots of plantain (Musa sp.) cv. Spambia (genome AAB) incubated on a semi-solid modified Murashige and Skoog (MS) medium supplemented with 4.5 μM
2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 μM zeatin. About 25% of shoot-tip explants formed scalps, and about 98% of
scalps developed embryogenic calli. Small dense aggregates of cells, were obtained when these calli were transferred to liquid
MS medium supplemented with 4.5 μM 2,4-D and 1.0 μM zeatin. Upon transfer to semi-solid MS medium of the same composition
as described above, aggregates of cells formed somatic embryos. In the presence of 2.5 μM abscisic acid (ABA), maturation
of somatic embryos was 2.6-fold higher than that of control (lacking ABA), and regardless of the type of cytokinin used in
the medium. Upon transfer to MS medium supplemented with 1.25 μM 6-benzyladenine (BA), 80% of germinated embryos developed
into plantlets. 相似文献
16.
Parthasarathi Bhattacharya Satyahari Dey Nilanjana Das Bimal Chandra Bhattacharyya 《Plant cell reports》1990,9(8):439-442
A procedure for rapid multiplication of Chrysanthemum morifolium RAMAT cv. Birbal Sahni using leaf callus and stem (nodal/internodal) callus as well as node and apical shoots has been developed. Murashige and Skoog's medium (1962) supplemented with 2mg/1 2,4-D yielded good green calli from both leaf and stem segments within 2 weeks. About 1 cm × 1 cm callus regenerated 2–3 shoots after 3 weeks on MS solid medium supplemented with 0.1 mg/l IAA and 0.2 mg/l BAP. Each of the regenerated shoots when transferred to the same shooting medium without agar yielded about 150 new shoots, which in turn regenerated roots after another week in MS half strength or modified White's media (Rangaswamy, 1961). It has been estimated that about 1014 plantlets could be produced in a year from one expiant following the proposed protocol.Abbreviations BAP
6-benzylaminopurine
- IAA
indole-3-acetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog's (1962) medium 相似文献
17.
Hypocotyl segments of 2- to 3-week-old radish (Raphanus sativus L. cv. F1 Handsome Fall) seedlings produced yellowish compact calli when cultured on Murashige and Skoog's (MS) medium supplemented with 1 mgl-1 2,4-dichlorophenoxyacetic acid (2,4-D). Upon transfer onto medium containing 6-benzyladenine and -naphthaleneacetic acid, up to 5.3% of the calli gave rise to a few somatic embryos. When subcultured for 3 to 6 months, 7% of the yellowish, compact calli produced white, compact calli which formed numerous embryos. These calli maintained their embryogenic capacity for over 18 months. When cultured on medium containing 0.1 to 3 mgl-1 2,4-D, up to 90% of longitudinally sliced somatic embryo halves produced calli with numerous secondary embryos. Embryos were transferred onto medium containing 0.1 mgl-1 2,4-D and 1 mgl-1 abscisic acid where they developed into the cotyledonary stage. Upon transfer onto half-strength MS basal medium, approximately 90% of the embryos developed into plantlets. These plantlets were successfully transplanted in potting soil and after cold treatment they were grown to maturity in a phytotron.Abbreviation 2,4-D
2,4-dichlorophenoxyacetic acid
- BA
6-benzyladenine
- GA3
gibberellin A3
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog
- NAA
-naphthaleneacetic acid 相似文献
18.
Slavica Ninković Tatjana Djordjević Branka Vinterhalter Branka Uzelac Aleksandar Cingel Jelena Savić Svetlana Radović 《Plant Cell, Tissue and Organ Culture》2010,103(1):81-91
Agrobacterium rhizogenes A4M70GUS-mediated transformation of two local breeding lines of sugar beet was obtained using 4-week-old seedlings. Root
formation efficiency was 61.54% for SBa genotype and 36.36% for SBb genotype. Five highly proliferated hairy root lines have
been established in liquid hormone-free MS medium. Transgenic nature of the hairy root clones was evaluated by GUS assay,
PCR and RT-PCR analyses. Hairy root-derived calli were induced using different plant growth regulators (PGRs): auxin, auxin/cytokinin
and cytokinin. The best callus induction response was achieved on MS medium containing both 1 mg/l 2,4-dichlorophenoxyacetic
acid (2,4-D) and 1 mg/l thidiazuron (TDZ). Globular embryo-like structures were observed in friable callus after its prolonged
cultivation on MS medium supplemented with TDZ and giberellic acid (GA3) at 1 mg/l each, followed by growth on MS medium containing 1% glucose and 0.5 mg/l 2,3,5-triiodobenzoic acid (TIBA). Histological
analysis revealed somatic embryos at different stages of development in hairy root-derived callus of sugar beet. 相似文献
19.
Lucia Martinelli Paola Bragagna Valentino Poletti Attilio Scienza 《Plant cell reports》1993,12(4):207-210
Somatic embryogenesis from leaf- and petiole-derived calli of Vitis rupestris was obtained with an efficiency of 3.2% and 4.2% of plated explants, respectively on two combinations of 6-benzyladenine and 2,4-dichlorophenoxyacetic acid (1/0.1 and 1/1 mgl–1) added to MS medium. Embryogenic callus, embryo subcultures and somatic embryogenesis from somatic embryos were obtained either in the presence of 1 mgl–1 indole-3-acetic acid or 0.1 mgl–1 indole-3-butyric acid added to MS or NN media. Within a 4-month culture, embryo germination occurred at a frequency of 13% of explanted embryos when chilling at 4°C was provided for two weeks and a combination of 6-benzyladenine (1 mgl–1) with indole-3-butyric acid (0.1 mgl–1) was added to NN medium supplemented with casein hydrolysate (250 mgl–1). A higher frequency (51%) was obtained in a longer culture time (9 months) when only indole-3-butyric acid was present in the medium and in absence of chilling.Abbreviations BA
6-benzyladenine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- MS
Murashige and Skoog (1962)
- NN
Nitsch and Nitsch (1969)
- NOA
2-naphthoxyacetic acid 相似文献
20.
A highly efficient tissue culture system and Agrobacterium-mediated transformation protocol for Chinese upland rice cultivar Handao297 has been established with mature embryos as explants.
Up to 81.2% of mature embryos were induced to regenerate good-quality calli on NB medium (a medium combining N6 macronutrient
components and B5 micronutrient and organic components) containing 3 mg/l 2,4-dichlorophenoxyacetic acid in 10 days. More
than 80% of the calli were morphogenic within 1 week and regenerated green plantlets within 1 month on Murashige and Skoog
medium supplemented with 0.5 mg/l 6-benzyladenine, 0.5 mg/l kinetin, 1 mg/l zeatin, 0.5 mg/l thidizazuron (TDZ), 0.5 mg/l
naphthaleneacetic acid, 0.15 mg/l indoleacetic acid, and 0.15 mg/l indolebutyric acid. This tissue culture system was suitable
for Agrobacterium-mediated transformation of upland rice Handao297. Furthermore, some important factors affecting transformation frequency
were investigated with Agrobacterium strain AGL1 containing the plasmid pCAMBIA1381. The addition of 30 mg/l hygromycin B followed by 60 mg/l hygromycin B to
the selection induction medium facilitated the revival of calli from selection and reduced false positive calli. Hygromycin
B at 10 mg/l was most effective in suppressing non-transgenic callus growth in the differentiation medium. The addition of
TDZ to the differentiation medium promoted the morphogenesis of calli and facilitated the generation of adventitious shoots
by five to tenfold in comparison to medium without TDZ. 相似文献