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1.
Summary Methods of plant regeneration from callus and protoplasts of Helianthus giganteus L. are described. Embryogenic callus was obtained from leaf explants and plants were regenerated from these calli on MS media with different combinations of benzyladenine and naphtaleneacetic acid. Leaf protoplasts isolated from in vitro grown plants formed somatic embryos when cultured in agarose solidified droplets of V-KM medium containing benzyladenine and naphtaleneacetic acid. Embryos developed into plantlets on media with reduced auxin contents. Regenerated plants were successfully planted in soil.Abbreviations BA
benzyladenine
- IAA
indoleacetic acid
- MS
Murashige and Skoog medium
- NAA
naphtaleneacetic acid
- V-KM
protoplast culture medium of Binding and Nehls 相似文献
2.
A sunflower genotype (Helianthus annuus L. cv. Florom-328) able to regenerate plants from in vitro cultures was identified by screening hybrids and inbred lines. Protoplasts of this genotype were isolated from dark grown hypocotyls and were cultured in droplets of agarose-solidified V-KM medium covered by liquid V-KM supplemented with naphthaleneacetic acid (NAA) and benzylaminopurine (BAP). One week later colonies were subjected to 2,4-dichlorophenoxyaceticacid for a one week period. Further culture in V-KM with reduced concentrations of NAA and BAP resulted in the appearence of somatic embryos. Maturation of embryos was achieved by culture on MS medium supplemented with NAA, BAP, gibberellic acid A3 and the ethylene inhibitor AgNO3. Embryos were then transferred onto hormone free MS medium for germination. The frequency of shoot formation in the best case was 9.6 percent of viable colonies (1.3 percent of protoplasts plated). Some of the shoots with roots could be transplanted into soil, others were grafted on hypocotyls of in vivo germinated seedlings. Eighty percent of grafted shoots and over 95 percent of rooted shoots survived. The plants flowered and produced 5 to 10 seeds each. Factors affecting the frequency of embryo formation and plant regeneration are discussed.Abbreviations BAP
6-benzylaminopurine
- GA3
gibberellic acid
- MES
morpholinoethanesulfonic acid
- MS
Murashige and Skoog medium
- NAA
naphthaleneacetic acid
- V-KM
protoplast culture medium of Binding and Nehls
- 2,4D
2,4-dichlorophenoxyacetic acid 相似文献
3.
A simple and efficient protocol for plant regeneration from mesophyll protoplasts ofNierembergia repens is described. The protoplasts divided in modified half-strength MS (/12 MS) medium containing benzylaminopurine (BA) and a-naphthaleneacetic acid (NAA) and formed visible colonies after 2 weeks which produced single adventitious shoots 4 weeks later. Plating efficiency (11.2%), percent colony formation (0.84%), and the number of shoot-forming colonies (368/dish) were highest in /12 MS containing 0.1 mg/l BA and 0.05 mg/l NAA. However, the percentage of colonies with shoot formation was highest (31.8%) in /12 containing 0.05 mg/l BA and 0.01 mg/l NAA. Almost all of the remaining colonies (97.5%) also regenerated shoots upon transfer onto MS medium containing 0.05 mg/l BA. The shoots with 2–3 leaves readily rooted 3–5 days after insertion in /12MS lacking plant growth regulators. Regenerated plantlets were easily established in soil 50 days after protoplast isolation. All the regenerants were normal and possessed diploid chromosome numbers.Abbreviations
BA
Benzylaminopurine
-
FDA
fluorescein diacetate
-
MES
2-(Morpholino)ethanesulfonic acid
- MS
Murashige and Skoog (1962) medium
- /12MS
half strength MS
- NAA
-naphthaleneacetic acid
-
PGR
plant growth regulator 相似文献
4.
Muhammad Akbar Anjum 《Acta Physiologiae Plantarum》1998,20(2):129-133
Freshly isolated mesophyll and suspensions-cell protoplasts of S. tuberosum cvs. Desiree and Maris Piper were cultured in different media i.e. modified MS, V-KM and MS-KM. Protoplast plating efficiencies
were higher in MS-KM medium. Resulting protoplast-derived calluses were transferred either onto the medium of Bokelmann and
Roest (1983) or that of Lam (1977) for shoot regeneration. Calluses derived from mesophyll cell protoplasts differentiated
about 2 weeks earlier than calluses derived from suspension-cell protoplasts. Shoot initiation was also about 2 weeks earlier
from calluses subcultured onto the former medium as compared to the latter. 相似文献
5.
Plant regeneration from mesophyll protoplasts of lisianthus (Eustoma grandiflorum) by adding activated charcoal into protoplast culture medium 总被引:1,自引:0,他引:1
Hisato Kunitake Toshiki Nakashima Kinya Mori Masanobu Tanaka Masahiro Mii 《Plant Cell, Tissue and Organ Culture》1995,43(1):59-65
Plant regeneration from isolated protoplasts of 8 cultivars of lisianthus, Eustoma grandiflorum (Griseb.) Schinners, has been established by using activated charcoal. Protoplasts were isolated from lisianthus leaves grown in vitro and started to divide within 3–4 days of culture, but successful colony formation was only achieved by adding gellan gum blocks containing 1% (w/v) activated charcoal immediately after culture. Colonies consisting of as many as 50–100 cells formed after 30 days of culture and were transferred to fresh medium for callus proliferation and shoot regeneration, respectively. These shoots rooted on MS medium containing 0.5 mg l–1 indolebutyric acid(IBA) and the plantlets were finally transplanted to pots. Morphological characteristics, growth habit and pollen fertility of protoplast-derived plants of one cultivar were not different from those of seed-grown plants as control.Abbreviations BA
6-benzylaminopurine
- NAA
1-naphthaleneacetic acid
- MS
Murashige & Skoog (1962) medium
- IBA
indolebutyric acid
- MES
2-N-morpholinoethane sulfonic acid 相似文献
6.
Zhao Yan-Xiu Philip J. C. Harris Yao Dun-Yi 《Plant Cell, Tissue and Organ Culture》1995,40(2):119-123
Protoplasts were isolated from cotyledons of Sesbania bispinosa (Jacq.) W.F. Wight. In a liquid-over-agar culture system with Murashige and Skoog (MS) medium supplemented with 1 mg l-1 2,4-dichlorophenoxyacetic acid (2,4-d, 2 mg l-1 benzyladenine (BA), 1 mg l-1 glutamine and 0.5 and formed callus. The first division occurred after 3–4 days. Callus formed from the protoplasts differentiated shoots by organogenesis on MS medium with 1 mg l-1 indolebutyric acid (IBA) and 1 mg l-1 BA. These shoots developed into complete plantlets when excised and cultured on MS medium with 0.5 mg l-1 IBA. 相似文献
7.
Kong-Nan Zhao Dennis J. Bittisnich Gerald M. Halloran Malcolm I. Whitecross 《Plant Cell, Tissue and Organ Culture》1995,40(1):73-84
Cotyledons from twelve cultivars of Brassica; B. napus (Westar, Eureka, Global, Pivot and Narc 82); B. campestris: (Arlo, Sonja, Bunyip and Wonk Bok) and B. oleracea (Phenomenal Early, Sugar Loaf and Earliball) were used for protoplast isolation and culture in a comparative study of cell colony and callus formation, and plant regeneration. The formation of cell colonies and callus from protoplast cultures were significantly influenced by the light conditions of seed germination. All twelve cultivars showed callus formation from protoplast cultures derived from cotyledons of seedlings grown in dark for 3 days followed by 1 day dim light (dark/dim light-grown). Callus was obtained in all five liquid media used: modified K8P(1), modified K8P(2), modified MS, modified B and modified NN. In contrast, only six cultivars exhibited callus formation from the protoplasts isolated from cotyledons of seedlings germinated under light conditions for 7 days (light-grown) and in only three media: modified K8P(1), modified MS, modified B.Callus, derived from protoplast cultures isolated from dark/dim light-grown cotyledons and grown on K3 or MS series solid media for about 1 month, could develop shoots when further transferred onto MS series regeneration media. All five cultivars of B. napus, three of the four cultivars of B. campestris (Arlo, Sonja and Bunyip) and one of the three cultivars of B. oleracea (Sugar Loaf) exhibited shoot regeneration from protoplast cultures within 2–3 months after protoplast isolation. The frequency of shoot regeneration ranged among 1–22.5%. A high degree of reproducibility was observed in cultivars Westar, Eureka, Global, Arlo, Bunyip and Sugar Loaf. In contrast, among the six cultivars that formed callus in protoplast culture derived from light-grown cotyledons, only three cultivars from B. napus (Westar, Eureka, Global) exhibited shoot regeneration 5.5 months after protoplast isolation. Regenerated shoots from cultivars Westar, Eureka and Bunyip and Sugar Loaf, which derived from protoplasts of dark/dim light germinated seedling and were induced to root on rooting media, survived in soil and grew to produce silique and set seeds.Abbreviations 2,4-d
2,4-dichlorophenoxyacetic acid
- BA
benzylaminopurine
- EDTA
ethylenediaminetetraacetic acid
- IAA
indole-3-acetic acid
- IBA
indole-3-butyric acid
- KT
kinetin
- GA3
gibberellic acid
- MS
Murashige and Skoog medium
- NAA
-naphthaleneacetic acid
- PAR
photosynthetically active radiation 相似文献
8.
S. J. Ochatt 《Plant Cell, Tissue and Organ Culture》1991,25(2):161-167
Leaf protoplasts of axenic shoot cultures of Lonicera nitida cv Maigrun underwent sustained division to give multicellular colonies (microcalli) on a modified, ammonium-free MS (Murashige & Skoog) medium containing 0.5 mg l-1 NAA (1-naphthaleneacetic acid), 1.0 mg l-1 BAP (6-benzylaminopurine) and 150 mg l-1 casein enzymatic hydrolysate. Callus was produced upon transfer of cell colonies to MS medium with 2.0 mg l-1 NAA and 0.2 mg l-1 BAP. About 110 days from isolation protoplast-derived shoots were regenerated on a half-strength MS medium with 0.01 mg l-1 NAA, 5.0 mg l-1 BAP, 0.5 mg l-1 zeatin and a complex mixture of group B vitamins. The replacement of such mixture by 250 mg l-1 casein enzymatic hydrolysate promoted rhizogenesis in calli, with shoot buds being subsequently regenerated from the protoplast-derived roots. Micropropagation of protoplast-derived shoots (of either origin) was difficult, due to a strong apical dominance, but could be accomplished by transferring single-node explants to half-strength MS medium with 1.5 mg l-1 BAP. Such shoots were, in turn, successfully rooted and transferred to the glasshouse where they completed acclimatization.Abbreviations BAP
6-benzylaminopurine
- CPW
Power et al. (1989) medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- FDA
fluorescein diacetate
- F.P.E.
final plating efficiency
- f.wt.
fresh weight
- IAA
4-indole-3yl-acetic acid
- IBA
4-indole-3yl-butyric acid
- I.P.E.
initial plating efficiency
- MES
2-N-morpholinoethane sulfonic acid
- M.P.E.
intermediate plating efficiency
- MS
Murashige & Skoog (1962) medium
- NAA
1-naphthaleneacetic acid
- PVP-10
polyvinylpirrolidone
- Av MW 10,000, TIBA
2,3,5-tri-iodobenzoic acid
- Z
zeatin 相似文献
9.
Callus cultures of Prosopis tamarugo Phil (Leguminosae, Sub family-Mimosoideae) were established from hypocotyls and cotyledons on MS medium supplemented with NAA (2.0 mg l-1) and BAP (0.2 mg l-1). Regeneration through various juvenile explants was obtained on hormone-free and high cytokinin containing Murashige and Skoog's medium. Multiple shoot buds formation was observed from the embryonic axis on MS medium incorporated with BAP (5.0 mg l-1)). Elongation of shoot buds was observed on subsequent transfer to MS medium with BAP (1.0–2.5 mg l-1) or without BAP. Explants containing apical meristem showed higher number of shoot formation at an early period. De novo shoot buds formation through callus morphogenesis was observed at the base of differentiated shoots on high cytokinin containing medium. All the manipulations of salt strength of MS, nitrogen, carbon, ascorbic acid and polyamines failed to induce organogenesis in isolated callus. In vitro produced shoots were rooted on MS medium supplemented with IBA or NAA singly or in combination.Abbreviations HC
high cytokinin (BAP 5.0 mg l-1)
- BAP
6-benzyl amino purine
- IBA
indole-3-butyric acid
- HF
hormone free
- NAA
I-naphthalene acetic acid
- MS
Murashige & Skoog 相似文献
10.
A. S. Kochevenko Y. I. Ratushnyak Y. Y. Gleba 《Plant Cell, Tissue and Organ Culture》1996,44(2):103-110
Mesophyll protoplasts of species of series Juglandifolia (Solanum rickii, S. lycopersicoides, S. ochranthum and S. juglandifolium) were isolated and cultured in liquid nutrient media TM-2 or KM8P. The cell colonies formed were transferred onto agar-solidified media TM-3 or GM, and 10 to 15 days later onto TM-4, PRM, MS3ZG, KK or C regeneration media. Formation of the shoots for S. rickii and S. lycopersicoides was observed after 30 to 35 days on regeneration medium. The regenerated shoots were rooted on hormone-free MS medium. Morphological and cytogenetic analyses have shown that somaclonal variants might arise in the course of plant regeneration from protoplasts of these species.Abbreviations BAP
6-benzylaminopurine
- 2, 4-D
2, 4 dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- GA3
gibberellic acid
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog medium
- B5
Gamborg medium
- TRIS
tris(hydroxymethyl)aminomethane 相似文献
11.
A tissue culture method is described for clonal multiplication of Leucaena leucocephala K67 using single lateral bud explants from 2–3 m tall greenhouse grown trees. N-6 benzyladenine (BA: 3.0 mg.1-1) and napthaleneacetic acid (NAA: 0.05 mg.1-1) in Murashige & Skoog's (MS) medium were found to be best suited for multiple shoot differentiation in 4–5 week old cultures.
Analysis of variance of the main treatment effects of BA and NAA on shoot parameters showed that BA significantly (P=0.001) affected shoot development while NAA did not. A shoot multiplication rate of 22±3.63 shoots per bud explant was obtained
in 150 days on 1/2 strength MS medium with 3.0 mg.1-1 BA and 0.05 mg.1-1 NAA. Shoots developed adventitious roots within 15 days in 1/2 strength MS medium containing indole-3-butyric acid (IBA:
3.0 mg.1-1) and Kinetin (0.05 mg.1-1). Eighty percent of the transplanted plantlets are being grown in greenhouse conditions. 相似文献
12.
Protoplasts isolated from both 7-day-old light-grown and 4-day-old dark/dim light-grown cotyledons of four Brassica campestris varieties (Arlo, Sonja, Bunyip and Wonk Bok) were cultured in three liquid media: modified K8P, modified MS and modified Pelletier's B to compare the capacities for cell division and plant regeneration. Following cell wall regeneration the cultured protoplasts from dark/dim light-grown cotyledons of four varieties showed rapid division and high frequency of cell division compared with those isolated from light-grown cotyledons. The frequencies of cell division were significantly influenced by varieties and culture media but only in cultured protoplasts isolated from dark/dim light-grown cotyledons. The interaction between varieties and media was also significant. Cell colonies formed within 7–14 days in protoplast cultures from dark/dim light-grown cotyledons, and calli subsequently grown on a solid medium developed shoots when transferred onto a regeneration medium. Three of four tested varieties (Arlo, Sonja and Bunyip) showed shoot regeneration within 2–3 months after protoplast isolation, with a high degree of reproducibility in Arlo and Bunyip. Regenerated shoots, which were induced to root on half-strength MS medium with 0.1 mg.l–1 IBA, survived in soil and grew to produce siliques and set viable seeds in the greenhouse. The present report is the first to document the production of regenerated plants that set seeds in Brassica campestris from cotyledonary protoplasts.Abbreviations BAP
benzylaminopurine
- CPW
Composition of Protoplast Washing-solution
- 2,4-D
2,4-dichlorophenoxyacetic acid
- EDTA
ethylenediamine-tetraacetic acid
- GA3
gibberellic acid
- IBA
indole-3-butyric acid
- IAA
indole-3-acetic acid
- MS
Murashige and Skoog medium
- NAA
-naphthaleneacetic acid
- KT
kinetin
- FDA
fluorescein diacetate
- SDS
sodium dodecyl sulfate 相似文献
13.
Cao Dinh Hung Krystyna Johnson Fraser Torpy 《In vitro cellular & developmental biology. Plant》2006,42(6):548-552
Summary An efficient protocol for in vitro propagation of the valuable medicinal plant, Wasabia japonica (Miq.) Matsumura is described through shoot tip proliferation and direct regeneration. Multiple shoots were induced from
shoort tips cultured on Murashige and Skoog (MS) semi-solid medium containing various concentrations (0.5–50 μM) of N6-benzyladenine (BA), thidiazuron, kinetin, and zeatin. A comparison was made on shoot multiplication between semi-solid and
liquid culture media. Well-developed shoots were obtained using full-strength MS semi-solid medium containing 5.0 μM BA. However, the greatest shoot proliferation was achieved on either full- or half-strength MS liquid media supplemented
with 5.0 μM BA for 4 wk (15.3±0.9 and 15.0±0.7 shoots per explant, respectively), and on half-strength MS liquid medium for 6 wk (25.8±1.3
shoots per explant) in culture. In contrast, the maximum number of shoots per explant on full-strength MS semi-solid medium
was achieved with either 5.0 μM BA (10.4±0.6 shoots per explant) or 10.0 μM kinetin (10.9±0.8 shoots per explant). Fresh weight of explants and length of shoots derived from full-strength MS liquid
medium (1055±77 mg and 34.2±1.0 mm, respectively) were significantly higher than those derived from full-strength MS semisolid
medium (437.6±17.3 mg and 15.4±0.7 mm, respectively). Quarter-strength MS liquid medium had no significant difference in shoot
proliferation when compared to quarter-strength MS semi-solid medium. Elongated shoots were separated and rooted on half-strength
MS semi-solid media fortified with 1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), or indole-3-acetic acid (IAA)
ranging from 0.1 to 10.0 μM. Root formation was greatest with IBA when compared with IAA and NAA. One hundred percent of shoots were rooted on half-strength
MS medium with 5.0 μM IBA, while vigorous roots were obtained with 10.0 μM IBA. Micropropagated plantlets were successfully established in soil with 95% survival rate after heardening. 相似文献
14.
Johannes Siemens María Torres Marion Morgner María Dolores Sacristán 《Plant cell reports》1993,12(10):569-572
A protocol for obtaining regenerated fertile plants from mesophyll protoplasts of four ecotypes (Col C24, Per-1, Bur-0, Landsberg erecta) and two marker lines (M4 and M10) of Ardbidopsis thaliana is described. The different lines showed plating efficiencies between 1.0 and 3.9% using Nitsch medium or this medium supplemented with coconut water. For the differentiation of callus into normal shoots a single shoot regeneration medium was applicable to all ecotypes, but depending on the line other regeneration media showed to be more suitable. The results indicated that the protoplast culture procedure is applicable, with minor modifications, to all tested genotypes but the most suitable shoot regeneration medium should be established for each A. thaliana line.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzyl-aminopurine
- IAA
indole-3-acetic acid
- IPA
isopentenyladenine
- IPAR
isopentenyladenosine
- MES
2-[N Morpholino]ethanesulfonic acid
- MS
Murashige and Skoog
- NAA
naphthaleneacetic acid 相似文献
15.
High-frequency bud break and multiple shoots were induced in apical shoot buds and nodal explants ofMorus cathayana, M. lhou andM. serrata on Murashige and Skoog (MS) medium containing 0.5–1.0 mg/l 6-benzylaminopurine (BAP). Addition of gibberellic acid (0.4 mg/l) along with BAP induced faster bud break both in apical shoot buds and nodal explants and also enhanced the frequency of bud break in all three species. Shoot culture initiation was greatly influenced by explant type, explant age and explanting season. The shoots were successfully rooted on half-strength MS medium containing a combination of indole-3-acetic acid, indole-3-butyric acid and indole-3-propionic acid, each at 1.0 mg/l. The plantlets were successfully acclimated and eventually established in soil.Abbreviations
BAP
6-Benzylaminopurine
-
GA
3
Gibberellic acid
-
IAA
Indole-3-acetic acid
-
IBA
Indole-3-butyric acid
-
IPA
Indole-3-propionic acid
-
Kn
Kinetin
-
MS
Murashige and Skoog (1962) medium
-
NAA
1-Naphthalene acetic acid 相似文献
16.
Summary Protoplasts of Helianthus giganteus and Helianthus annuus were fused using polyethylene glycol. Before fusion H.giganteus protoplasts were subjected to iodoacetic acid treatment to render them unable to divide. Fused protoplasts were cultured in V-KM medium containing benzylaminopurine and naphtaleneacetic acid. Hybrid calli were identified on the basis of their ability of embryogenic development contributed by the Helianthus giganteus parent. Fifty embryogenic calli were cultured on MS based medium without growth regulators to induce further development of somatic embryos. Elongated shoots were removed, rooted and transferred into growth chambers. Overall morphology of the plants was intermediate between the two parents. Their hybrid nature was confirmed by chromosome counting and by the analysis of esterase isozymes. The plants flowered within two to three months and later died. Thus the perennial nature of H.giganteus is a recessive trait in this interspecific hybrid. Seeds were obtained from two of the regenerated plants. From these seeds normal fertile F2 plants could be grown.Abbreviations PEG
polyethylene glycol
- NAA
naphtaleneacetic acid
- BA
benzyladenine
- MS
Murashige and Skoog medium
- V-KM
protoplast culture medium of Binding and Nehls 相似文献
17.
In vitro induction of multiple shoots and plant regeneration in cotton (Gossypium hirsutum L.) 总被引:5,自引:0,他引:5
D. C. Agrawal A. K. Banerjee R. R. Kolala A. B. Dhage A. V. Kulkarni S. M. Nalawade S. Hazra K. V. Krishnamurthy 《Plant cell reports》1997,16(9):647-652
Induction of multiple shoots in cotton (Gossypium hirsutum L. cv. Anjali-LRK 516) has been achieved with cotyledonary nodes devoid of cotyledons and apical meristems. Explants from 35-day-old seedlings yielded the maximum number of shoots (4.7 shoots/explant) using Murashige and Skoog (MS) basal medium supplemented with 6-benzylaminopurine and kinetin (2.5 mg/1 each). Explants from 35-day-old seedlings raised in glass bottles produced a higher number of multiple shoots (8.3 shoots/explant) than those grown in glass tubes and cultured on the same shoot induction medium. Elongation of multiple shoots was obtained on liquid or agar MS basal medium without phytohormones. In vitro shoots were rooted on half-strength agar-solidified MS basal medium or with 0.05 or 0.1 mg/1 naphthaleneacetic acid. Hardening and survival of tissue culture plantlets was 95% under greenhouse conditions.Abbreviations
BAP
6-Benzylaminopurine
- GA3
Gibberellic acid
- MS
Murashige and Skoog medium
-
NAA
-Napthaleneacetic acid 相似文献
18.
C. L. Webb M. R. Davey J. A. Lucas J. B. Power 《Plant Cell, Tissue and Organ Culture》1994,38(1):77-79
Cultured protoplasts of young, unexpanded leaves of the wild lettuce, Lactuca perennis, divided to produce cell colonies in an agarose-solidified, modified MS medium with reduced levels of inorganic salts, together with 2,4-d, NAA and zeatin at 0.2, 0.1 and 0.5 mg 1-1 respectively. Organogenesis followed the initial transfer of protoplastderived colonies to modified MS medium with 2,4-d, NAA and zeatin (0.1, 1.0 and 0.2 mg 1-1 respectively) and then to full-strength MS medium with 6-BA and NAA (0.4 and 0.05 mg 1-1). Shoots were rooted on agar-solidified MS medium lacking growth regulators. Regenerated shoots were established ex vitro, 21 weeks after protoplast isolation.Abbreviations 6-BA
6-benzyladenine
- BSA
bovine serum albumin
- d
days
- 2.4-d
2,4-dichlorophenoxyacetic acid
- f. wt.
fresh weight
- IAA
indoleacetic acid
- MES
2 [N-morpholino]ethane sulphonic acid
- MS
Murashige & Skoog (1962) 相似文献
19.
N. S. Shekhawat T. S. Rathore R. P. Singh N. S. Deora S. R. Rao 《Plant Growth Regulation》1993,12(3):273-280
Genotype, age of tree, nature of explant and size (length and diameter), season of explant collection, explant position on medium, plant growth regulators and certain additives (ascorbic and citric acids, adenine sulphate, L-arginine, glutamine and ammonium citrate), incubation conditions, and subculturing period greatly influenced the in vitro clonal propagation of P. cineraria. The maximum number of 10–12 shoots were induced from the nodal shoot segment from pruned thorny adult trees on Murashige and Skoog's (MS) medium containing 0.1 mgl-1 indole- 3-acetic acid (IAA)+2.5 mgl-1 benzylaminopurine (BAP)+additives. Higher temperature (31+-2°C) and mixed (fluorescent and incandescent) light of 50 mol m-2 s-1 photon flux density for 12 h per day photoperiod favoured shoot induction and subsequent growth. Explants from thornless trees produced 6–8 shoots per explant on MS medium containing 0.1 mgl-1 IAA+5.0 mgl-1 BAP + additives. Nodal shoot segments obtained from root and stump sprouts produced multiple shoots. Root segments differentiated into multiple shoots on MS medium containing 0.5 mgl-1 indolebutyric acid (IBA)+2.5 mgl-1 BAP.Differentiated shoots multiplied best on MS medium containing 0.1 mgl-1 naphthalene acetic acid (NAA)+1.0 mgl-1 BAP + additives. To yield multiple shoots the original explant was transferred 6 times on fresh medium after harvesting the differentiated shoots. Shoots were rooted by pulsing with 100 mgl-1 IBA for 4 h and then culturing on hormone-free half strength MS medium. Initial dark incubation for 5 days at high temperature (33±2°C) was found essential for root induction from shoots which was 63% within two weeks. The rooted plantlets contained a consistent number of chromosomes (2n=28). It is suggested that the protocol developed could be useful for cloning of mature and tested trees of P. cineraria. 相似文献
20.
Sarowar S. Oh H.Y. Hyung N.I. Min B.W. Harn C.H. Yang S.K. Ok S.H. Shin J.S. 《Plant Cell, Tissue and Organ Culture》2003,75(2):179-182
An efficient in vitro micropropagation protocol was developed for direct shoot growth of interspecific Cucurbita hybrid variety using shoot–tips of 5-day-old explants. The excised shoot–tips were cultured on Murashige and Skoog (MS) medium containing two plant growth regulators (6-benzyladenine and naphthaleneacetic acid) with various combinations and concentrations for the study of shoot induction. The best condition for shoot growth was with 3 mg l–1 6-benzyladenine (BA) in MS medium. The shooting frequency was 84% and five shoots were obtained from each explant after 30 days of culture. Shoots (11.5 cm length) were rooted most effectively in 1 mg l–1 indole-3 butyric acid (IBA)-supplemented MS medium. The highest root formation rate was 93% and all rooted shoots were transplanted into soil. 相似文献