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1.
M. S. Brar J. M. Al-Khayri C. E. Shamblin R. W. McNew T. E. Morelock E. J. Anderson 《In vitro cellular & developmental biology. Plant》1997,33(2):114-118
Summary Shoot multiplication was induced in cowpea, cv. Georgia-21, from shoot tip explants. Shoot tips, 5 mm long, were isolated
from in vitro-grown seedlings and cultured on MS medium containing N6-benzyladenine (BA) at 1, 2.5, or 5 mg/liter (4.4, 11.1, or 22.2 μM) or 6-furfurylaminopurine (kinetin) at 1, 2.5, or 5 mg/liter (4.6, 11.6, or 23.2 μM) combined with 2,4-dichlorophenoxyacetic acid (2,4-D) at 0.01, 0.1, or 0.5 mg/liter (0.05, 0.5, or 2.3 μM) or naphthaleneacetic acid (NAA) at 0.01, 0.1, or 0.5 mg/liter (0.05, 0.5, or 2.7 μM). Cultures were maintained at a 12-h photoperiod (40 μmol·m−2·s−1) and 23 ± 2° C. Treatments with BA induced greater shoot proliferation than those with kinetin. The highest number of shoots
was produced on 5 mg (22.2 μM) BA per liter in combination with NAA or 2,4-D at 0.01 mg/liter (0.05 μM). Callus proliferated from the basal ends of shoot pieces in all treatments. The cultures also formed roots in the presence
of kinetin, but not on BA-containing medium. To produce whole plants, the shoots were separated and rooted on 0.1 mg (0.5
μM) NAA per liter. Resulting plants grew normally under greenhouse conditions. Shoot tips provide an excellent explant source
for cowpea micropropagation and can be used for callus induction. 相似文献
2.
This report deals with micropropagation of the critically endangered and endemic Turkish shrub, Thermopsis turcica using callus, root and cotyledonary explants. Callus cultures were initiated from root and cotyledon explants on MS medium
supplemented with 0.5–20 μM NAA or 2,4-D. The root explants were found to be better in terms of quick responding and callusing
percentages as compared to the cotyledons. Organogenic callus production with adventitious roots and shoots were obtained
on MS medium with only NAA. The calli obtained with NAA, root and cotyledonary explants were cultured with BA and kinetin
(2–8 μM) alone or in combination with a low level (0.5 μM) of 2,4-D or NAA. The best regeneration of shoots from root explants
was observed on hormone-free MS medium. NAA with BA or kinetin in the medium improved shoot induction from the calli obtained
with NAA. Maximum percentage of shoots (93.3%), maximum number of shoots (6.2) and maximun length of shoots (8.22 cm) were
achieved from cotyledonary explants at 4 μM BA and 0.5 μM NAA. The presence of 0.5 μM or higher levels of 2,4-D in shoot induction
medium inhibited the regeneration in T. turcica explants. 83% of in vitro rooting was attained on pulsed-IBA treated shoots. The regenerated plants with well developed shoots and roots were successfully
acclimatized. Application of this study’s results has the potential to conserve T. turcica from extinction. 相似文献
3.
Luis Pedro Barrueto Cid Rolf Dieter Illg Aquiles E. Piedrabuena 《In vitro cellular & developmental biology. Plant》1994,30(3):150-155
Summary Aiming at the genetic improvement of garlic cultivars, a cell suspension protocol was established which includes the induction
of friable callus, establishment of cells in liquid medium, plating, regeneration, and bulb formation. Calluses of various
textures from compact to friable and from green to yellowish were obtained by culturing explants excised from inner leaves
of garlic bulbs on Marashig-Shoog (MS) medium with 2,4 dichlorophenoxy acetic acid (2,4-D), (1.1 mg/liter [5.0 μM]), picloram
(1.2 mg/liter [5.0 μM]), and kinetin (2.1 mg/liter [10 μM]). Friable callus occurred on MS-A contained 2,4-D alone (1.0 mg/liter
[4.52 μM]) and this callus was used to develop cell suspension cultures, which were maintained in liquid MS-B medium with a 2,4-D/benzyl
adenine (BA) (0.5 mg/liter [2.25 μM]: 0.5 mg/liter [2.22 μM]) ratio. High plating efficiency was obtained on MS-C medium with different naphthalene acetic acid/BA combinations. Regeneration
occurred after transfer of the caulogenic mass to MS-C medium containing 10 mg/liter (74.02 μM) and 20 mg/liter (148.04 μM) adenine for 60 days, followed by transfer to adenine-free medium. Plantlets transplanted to soil showed normal phenology.
Shoots grown on modified MS medium supplemented with indolylbutryic acid (3.0 mg/liter [14.7 μM]) stimulated bulb formation by 30 days in culture. 相似文献
4.
Tissue culture and plant regeneration of blue grama grass, Bouteloua gracilis (H.B.K.) Lag. Ex Steud
Gerardo Armando Aguado-Santacruz José Luis Cabrera-Ponce Víctor Olalde-Portugal M. A. Rosario Sánchez-González Judith Márouez-Guzmán Luis Herrera-Estrella 《In vitro cellular & developmental biology. Plant》2001,37(2):182-189
Summary As a first step towards applying biotechnology to blue grama, Bouteloua gracilis (H. B. K.) Lag. ex Steud., we have developed a regenerable tissue culture system for this grass. Shoot apices were isolated
from 3-d-old seedlings and cultured in 15 different growth regulator formulations combining 2,4-dichlorophenoxyacetic acid
(2,4-D), Picloram (4-amino-3, 5,6-trichloropicolinic acid), N6-benzyladenine (BA) or adenine (6-aminopurine). The highest induction of organogenic callus was obtained with formulations
containing 1 mg l−1 (4.52 μM) 2,4-D plus 0.5 mg l−1 (2.22 μM) BA. and 2 mg l−1 (8.88 μM) BA plus 1 mg l−1 (4.14 μM) Picloram with or without 40 mg l−1 (296.08 μM) adenine. Lower frequencies of induction were obtained for embryogenic as compared to organogenic callus. The most efficient
treatments for induction of embryogenic callus contained 2 mg l−1 (9.05 μM) 2,4-D combined with 0.25 (1.11 μM) or 0.50 mg l−1 (2.22 μM) BA, or 1 mg l−1 (4.52 μM) 2,4-D with 0.50 mg l−1 (2.22 μM) BA. Regeneration was achieved in hormonefree Murashige anmd Skoog (MS) medium, half-strength MS medium or MS medium plus
1 mg l−1 (1.44 μM) gibberellic acid. The number of plantlets regenerated per 500 mg callus fresh weight on MS medium ranged from 9 for 2 mg
l−1 (9.05 μM) 2,4-D to 62.2 for induction medium containing 2 mg l−1 (8,28 μM) Picloram, 1 mg l−1 (4.44 μM) BA and 40 mg l−1 (296.08 μM) adenine. Regnerated plants grown in soil under greenhouse conditions reached maturity and produced seeds. 相似文献
5.
Piedad Gallego Oscar Hita Nieves Villalobos Ana Dorado Luisa Martin Hilario Guerra 《In vitro cellular & developmental biology. Plant》2001,37(2):199-203
Summary An efficient plant regeneration system employing cotyledons, hypocotyls, petioles and leaves as explants and characterized
by continuous and prolific production of somatic embryos, has been developed with Medicago arborea ssp. arborea. The optimal somatic embryogenic response was obtained using a two-step protocol, where explants were incubated under a 16
h photoperiod for 2 mo. on Murashige and Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D; 9 μM) and kinetin (9 μM), and followed by transfer to kinetin-free MS medium with 2,4-D (2.25 μM). Removal of the cytokinin and a reduction in the concentration of auxin (2.25 μM) in the second step of culture were critical for enhanced production of somatic embryos. The best explants proved to be cotyledons
and petioles (i.e. a mean of 18.0±0.70 somatic embryos at 3 mo. for petiole culture). Somatic embryos were converted into
normal plantlets (8.0±0.89%) when cultured on basal MS medium with 5 μM indolebutyric acid. No somatic embryos were obtained when thidiazuron was used in the culture media. Using petioles as explants
and N6-benzyladenine (BA), embryogenesis was induced in the second step of culture when BA was removed from the medium and the concentration
of 2,4-D was decreased to 2.25 μM. 相似文献
6.
The role played by plant growth regulators in algae is poorly known. In order to increase the knowledge about the function of auxins and cytokinins in seaweeds, explants such as apical and intercalary segments and callus-like structures (CLS) of Grateloupia dichotoma were cultured in semi-solid or liquid artificial media ASP 12-NTA. Two auxins, indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), and one cytokinin, 6-benzylaminopurine (BA), at concentrations of 0.5 and 5.0 mg l–1 were tested. Moreover, IAA and BA were tested together at concentrations of 1:5 and 5:1 mg l–1. All treatments promoted the growth of CLS in intercalary segments; CLS from apical segments were significantly higher in treatments with 2,4-D or IAA:BA (1:5 mg 1–1). The morphogenetic responses for auxins and BA were opposite, auxins inhibited while BA promoted the formation of lateral branches; however, auxins promoted the elongation of such branches. The process of plant regeneration observed on CLS was stimulated significantly by treatment with high concentration of BA or IAA:BA (1:5 mg 1–1) in semi-solid and liquid media. The growth of upright axes was stimulated significantly by treatment with 2,4-D in semi-solid medium, and IAA:BA (1:5 mg l–1) in liquid medium. These results show the importance that plant growth regulators could have in the control of growth, morphogenetic processes and micropropagation in red algae.This paper is part of the PhD thesis of NSY. 相似文献
7.
Summary High-frequency somatic embryogenesis and plant regeneration was achieved on callus derived from leaf (petiole and lamina)
and internode explants of Centella asiatica L. Growth regulators significantly influenced the frequency of somatic embryogenesis and plant regeneration. Calluses developed
on Murashige and Skoog (MS) medium fortified with 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D) or 5.37 μM α-naphthaleneacetic acid (NAA), both with 2.32 μM kinetin (Kn), were superior for somatic embryogenesis. Callus developed on NAA and Kn-supplemented medium favored induction
and maturation of embryos earlier compared to that on 2,4-D and Kn. Embryogenic callus transferred from NAA and Kn-supplemented
medium to suspension cultures of half-strength MS medium with NAA (2.69 μM) and Kn (1.16 μM) developed a mean of 204.3 somatic embryos per 100 mg of callus. Embryogenic callus transferred from 2,4-D and Kn subsequently
to suspension cultures of half-strength MS medium with 2,4-D (0.45 μM) and Kn (1.16 μM) developed a mean of 303.1 embryos per 100 mg of callus. Eighty-eight percent of the embryos underwent maturation and conversion
to plantlets upon transfer to half-strength MS semisolid medium having 0.054 μM NAA with either 0.044 μM BA or 0.046 μM Kn. Embryo-derived plantlets established in field conditions displayed morphological characters identical to those of the
parent plant. 相似文献
8.
Myoung K. Kim Harry E. Sommer Jeffrey F. D. Dean Scott A. Merkle 《In vitro cellular & developmental biology. Plant》1999,35(1):37-42
Summary A sweetgum (Liquidambar styraciflua) nodule culture system was developed and integrated with genetic transformation by microprojectile bombardment. Nodule cultures
were established from seedling hypocotyls and proliferated in liquid medium containing 0.1 mg (0.45 μM) thidiazuron (TDZ) per 1 and 0.01 mg (0.045 μM) 2,4-dichlorophenoxyacetic acid (2,4-D) per 1. Shoots differentiated from the nodules in liquid media containing (per 1)
1 mg (4.4 μM) benzyladenine (BA), 0.5 mg (2.2 μM) BA, and 0.01 mg (0.054 μM) naphthaleneacetic acid (NAA), or 0.5 mg BA, 0.01 mg NAA, and 0.05 mg (0.23 μM) TDZ under the light. Differentiating shoots required 4 wk of dark treatment for further development on semisolid medium
containing 1 mg BA per 1. Elongated shoots were harvested and the basal ends were soaked in a solution containing 10 mg (49.2
μM) indole-3-butyric acid (IBA) per 1 before being planted in potting mix for ex vitro rooting. Roots formed and leaves expanded
in 2 wk. Sweetgum nodules were stably transformed by microprojectile bombardment with a 7.4-kb plasmid, pTRA 140, harboring
CaMV 35S-HPH and CaMV 35S-GUS. Evidence that nodules growing in the presence of hygromycin B were stably transformed was provided
by polymerase chain reaction analysis and β-glucuronidase activity. Sweetgum shoots differentiated in liquid medium in the
presence of hygromycin B. Shoots transferred to solid medium lacking hygromycin B elongated and displayed β-glucuronidase
activity in their expanding leaves and stems. Southern analysis confirmed the presence of the GUS gene in nodules and shoots.
Transgenic shoots initiated roots and showed leaf expansion 2 wk after being planted in potting mix. 相似文献
9.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
10.
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
11.
Eugenio Pérez Molphe Balch Martha E. Pérez Reyes Enrique Villalobos Amador Ernestina Meza Rangel Leticia del Rocío Morones Ruiz Hugo J. Lizalde Viramontes 《In vitro cellular & developmental biology. Plant》1998,34(2):131-135
Summary We have developed micropropagation systems for 21 species of Mexican cacti using explants from seedlings germinatedin vitro or shoot segments of juvenile 2–3-yr-old greenhouse plants. The species propagated belong to the generaAstrophytum, Cephalocereus, Coryphantha, Echinocactus, Echinocereus, Echinofossulocactus, Ferocactus, Mammillaria, Nyctocereus, andStenocactus. Multiple shoot formation from areoles was achieved in Murashige and Skoog (MS) medium supplemented with either 1 or 2 mg
N6-benzyladenine (BA) per 1 (4.44 or 8.87 μM) or BA at 1 or 2 mg/l plus naphthaleneacetic acid at 0.1 or 1 mg/l (0.54 or 5.37 μM). The requirements of growth regulators for optimal shoot proliferation, the velocity of the response, and the number of
buds produced by explant were different among the genera and species studied. Rooting of the shoots generatedin vitro was achieved in MS medium supplemented with indoleacetic acid at 0.5–1 mg/l (2.85–5.71 μM) or indolebutyric acid at 0.5–1 mg/l (2.46–4.90 μM). Finally, 70–95% of the rooted plants transferred to potting medium survived. 相似文献
12.
Summary Mature embryo axes of the Ohio buckeye were germinated on a medium containing 1 mg gibberellic acid (GA) per 1. Three wk following
germination, stem, petiole, and leaf blade tissues were excised and placed on media containing either 1 mg (4.5 μM) 2,4-dichlorophenoxy acetic acid (2,4-D) per 1, 1 mg (4.7 μM) kinetin per 1, 1 mg of both 2,4-D (4.5 μM) and kinetin (4.7 μM per 1, or 2 mg of both 2,4-D (9.1 μM) and kinetin (9.3 μM) per 1. Embryogenic tissue was formed only from stem segments after 2–3 mo. of culture on media containing both 2,4-D and
kinetin. Embryogenic tissue could be either maintained on solid medium for proliferation of embryogenic callus or placed in
liquid medium for proliferation of embryogenic suspension cultures. For transformation of suspension cultures, tissues were
inoculated with Agrobacterium EHA105 containing the binary plasmid Vec035, briefly sonicated, and cultured in the presence of 100 μM acetosyringone for 2 d. To eliminate Agrobacterium, tissues were washed and placed in liquid proliferation medium containing either 500 mg Cefotaxime per 1 or 400 mg TimentinŖ
per 1. Selection on 20 mg hygromycin per 1 was initiated 2 wk after inoculation, and after an additional 10 wk, hygromycin-resistant
tissue was isolated and separately cultured. Although some hygromycinresistant clones were recovered with no sonication treatment,
four to five times more clones were obtained following sonication. Putative transformed clones were confirmed to be transgenic
via both histochemical β-glucuronidase (GUS) assay and southern hybridization analyses. Development of transgenic embryos
occurred on a growth regulator-free medium containing 3% sucrose. After 2 mo. of embryo development, the embryos were transferred
to fresh medium for germination. 相似文献
13.
Callus-mediated and direct protocorm-like body formation of Bletilla striata and assessment of clonal fidelity using ISSR markers 总被引:3,自引:0,他引:3
Two efficient morphogenetic pathways for micropropagation of Bletilla striata (Thunb.) Reichb. f. have been established through the callus-mediated and direct formation of protocorm-like bodies (PLBs) from protocorms and shoot tips. Green calli were induced from the basal surface of protocorms and the cut-end of shoot tips on Vacin and Went (VW) medium supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or α-naphthalene acetic acid (NAA) after 3–5 weeks, with the highest frequency of explants forming callus (48.0 %) from protocorms at 1.0 mg l?1 2,4-D. The calli obtained from all plant growth regulator (PGR) treatments could proliferate and differentiate PLBs on the PGR-free medium. NAA and 2,4-D significantly enhanced the growth of callus. The fastest growth rate of callus was achieved at the combination of 1.0 mg l?1 2,4-D and 1.0 mg l?1 TDZ with 46.2-fold within 3 months. The regeneration of PLBs from callus was significantly improved by 6-benzyladenine (BA), and a mean number of 48.4 PLBs was produced from 100 mg calli at 1.0 mg l?1 BA within 3 months. BA and thidiazuron (TDZ) promoted the direct formation of PLBs from explants. The highest frequency of direct PLBs formation (76.0 %) and the highest mean number of PLBs per explant (30.2) were observed in protocorms cultured with 0.5 mg l?1 BA. Assessment of clonal fidelity by inter-simple sequence repeat (ISSR) markers revealed similarity ranges of 99.8–100.0 % between the regenerants and their mother plants and 99.5–100.0 % among the regenerants, which suggested the micropropagation protocols were genetically stable. 相似文献
14.
The micropropagation of adult Cleistanthus collinus was accomplished. The nodal segments from terminal twigs of a 15-year-old tree and basal sprouts of a comparable chronological
age were used for initiating shoot bud cultures. Washing explants with sterile mixtures of citric acid (520.5 μM) and PVP 40 (3.75 μM) three to four times controlled the leaching of brown inhibitory substances into the establishment medium. Axillary shoots
proliferated best on MS medium containing citric acid (104.1 μM), and PVP 40 (12.5 or 25 μM) supplemented with 0.44 μM BA. The number of new shoots from nodal segments of explants placed on MS medium supplemented with 0.44 μM BA increased when the remaining lengths of nodal segments were transferred to fresh medium after the longer microshoots were
harvested. The microshoots derived from basal sprouts rooted best (50%) when treated with 11.4 mM IAA for 2 min, whereas only 40% of the microshoots derived from terminal twigs produced roots after a 2-min exposure to 28.5
mM IAA. The placement of BA-soaked agar cubes on the apex-decapitated shoots controlled shoot-tip necrosis considerably. In
general, explants from basal sprouts were more suitable than terminal twig explants for the micropropagation of adult trees
of C. collinus.
Received: 26 February 1997 / Revision received: 13 September 1997 / Accepted: 29 September 1997 相似文献
15.
Summary Direct somatic embryo formation and plantlet regeneration was achieved from immature leaflets of chickpea (Cicer arietinum L.). Optimal somatic embryogenesis was obtained when immature leaflets were exposed to media supplemented with 15 μM 2,4-dichlorophenoxyacetic acid (2,4-D) for 7 d, to 2000 μM 2,4-D for 3 d, and to 50 μM 2,4-D for 10 d, followed by transfer onto Murashige and Skoog (MS) basal medium. Exposure of explants to high 2,4-D levels
(200–2000 μM) for 3 d produced bottle-shaped embryos, while exposure to low 2,4-D levels (<50 μM) and 50–2000 μM for 10 d produced spherical-shaped embryos. Two percent of embryos converted into plants upon culture on MS medium containing
15 μM gibberellic acid and 1 μM 3-indolebutyric acid. All regenerated plants were phenotypically normal. 相似文献
16.
Callus induction and micropropagation improved by colchicine and phytoregulators in Kappaphycus alvarezii (Rhodophyta, Solieriaceae) 总被引:1,自引:0,他引:1
Leila Hayashi Nair S. Yokoya Daniela M. Kikuchi Eurico C. Oliveira 《Journal of applied phycology》2008,20(5):653-659
Tissue culture techniques were applied for micropropagation of the red alga Kappaphycus alvarezii in order to select the best strain and experimental system for in vitro culture. Five strains were tested: brown (BR), green (GR) and red (RD) tetrasporophytes, brown female gametophyte (BFG),
and a strain originating from tetraspore germination (“Edison de Paula”, EP). The effects of three culture media were tested
on callus formation, regeneration from explants and from callus in the three tetrasporophytic and EP strains: seawater enriched
with half-strength of von Stosch’s (VS 50) and Guillard & Ryther’s (F/2 50) solutions, plus synthetic ASP 12-NTA medium, with
or without gelling agent. Explants of the EP strain were treated with glycerol and the phytoregulators indole-3-acetic acid
(IAA); 2,4-diclorophenoxyacetic acid (2,4-D); and benzylaminopurine (BA), alone or in combination. The effects of colchicine
(0.01%) during 24, 48, 72 hours and 14 days were analyzed in the BFG and EP strains. The EP strain showed the highest percentage
of explants forming callus and regeneration from explants in VS 50, indicating its high potential for micropropagation in
comparison to the other strains. Regeneration from callus was very rare. Treatments with glycerol and IAA:BA (5:1 mg L−1) stimulated the regeneration from explants. Significant differences were observed in the percentages of regeneration of EP
strain explants treated with colchicine for 14 days. Our results indicate that IAA and BA stimulated the regeneration process,
and that colchicine produced explants with high potential for regeneration, being useful for improving the micropropagation
of K. alvarezii. 相似文献
17.
An efficient tissue culture system for high frequency of plant regeneration from hypocotyl explants of Brassica carinata was developed via manipulation of culture medium and selection of explants. Explants grown on medium containing combinations of 2 mg l-1 BA and 0.01 mg l-1 NAA or 4 mg l-1 kinetin and 0.01 mg l-1 2,4-D regenerated shoots at 100% frequency. High frequency shoot regeneration occurred only from explants originating from 6 to 7-day-old but not younger or older seedlings. Explants showed higher regeneration capacity at the distal end than the proximal end, and the upper segment was more regenerative than the lower segment of hypocotyl. Regenerants were rooted on half-strength growth regulator-free medium, acclimatized and developed into normal, fertile plants.Abbreviations BA
benzyladenine
- 2-4-D
2,4-dichlorophenoxyacetic acid
- NAA
naphthaleneacetic acid
- MS
Murashige & Skoog 相似文献
18.
A. Sieboldianus (5-leaf aralia) is recalcitrant for micropropagation, but has very good landscaping potential. This research was conducted
with the following objectives: (1) to study effects of BA, TDZ, CPPU, 2iP, kinetin and zeatin in woody plant medium on the
performance of softwood shoot nodal explants produced by field grown 5-leaf aralia plants; (2) to investigate influences of
BA or TDZ in the forcing solution on subsequentin vitro shoot initiation of nodal explants taken from forced softwood growth. Shoot initiation of softwood nodal explants from field-grown
plants was promoted by adding BA, TDZ or CPPU to the culture medium. Kinetin, zeatin and 2iP were ineffective for micropropagation
ofA. Sieboldianus. The forced softwood growth for use as explants was “primed” by forcing dormant stems in solution containing 200 mg 8-HQC
per liter plus 2% sucrose, 44.4, 222, or 444 μM BA, or 45.4, 227, or 454 μM TDZ. BA and TDZ in the forcing solution enhanced
subsequentin vitro axillary shoot initiation of nodal explants taken from forced stems by doubling the number of shoots produced per explant
to 3.3 from 1.65 shoots per explant taken from field grown plants. This forcing solution technique also reduced the time needed
from culture initiation to potted plants to half of the time needed for the conventional micropropagation method (12 to 14
vs. 25 to 27 weeks), thus expediting the micropropagation ofA. Sieboldianus. 相似文献
19.
K. P. Martin A. Shahanaz Beegum C.-L. Zhang A. Slater P. V. Madhusoodanan 《Biologia Plantarum》2007,51(4):769-772
In vitro propagation of an anticancerous drug synthesizing plant, Ophiorrhiza prostrata D. Don, was established through indirect somatic embryogenesis. Friable embryogenic calluses were initiated from O. prostrata leaf and internode explants on Murashige and Skoog (MS) media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) either
alone or in combination with N6-benzyladenine (BA) or kinetin (KIN). Somatic embryos were developed after subculture of the friable calluses onto half strength
MS media containing 0.45 or 2.26 μM 2,4-D alone or in combination with BA or KIN. Medium supplemented with 2.26 μM 2,4-D and
2.22 μM BA was optimal, supporting the production of a mean of 5.8 globular embryos. Subculture of globular embryo-bearing
calluses on half strength MS medium without growth regulators produced the highest embryo frequency, and the majority of them
developing to early torpedo stage. Somatic embryos underwent maturation and converted to plantlets at high frequency (90 %)
on half strength MS medium supplemented with 0.44 μM BA. Somatic embryo-derived plantlets with well-developed roots were established
in field conditions with a 90 % survival rate. 相似文献
20.
Summary A protocol for micropropagation of plants via axillary bud proliferation from nodal explants of Terminalia bellirica Roxb. seedlings has been established. Explants were cultured on Murashige and Skoog medium with different concentrations
of 6-benzyladenine (BA; 4.4, 8.9, 13.3, 17.8, or 22.2 μM) or kinetin (Kn; 4.6, 9.3. 14.0, 18.6, or 23.2 μM). Within the range evaluated, the medium containing 13.3 μM BA showed the highest shoot length (1.9=0.2 cm) in the primary culture. When separated and transferred to fresh subculture
medium with lower levels of BA (2.2. 4.4, 6.6, or 8.9 μM) or Kn (2.3, 4.6, 6.9, or 9.3 μM), the nodal segments from individual regenerants (obtained initially from seedling nodes) showed efficient shoot induction
at 4.4 μM BA. Rooting of the shoots was achieved under in vitro conditions on two media tested, i.e., modified Gamborg's (B5) medium or Woody Plant Medium, both supplemented with 4.9 μM indole-3-butyric acid. Regenerated plants were established in the greenhouse. 相似文献