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1.
In vitro shoot regeneration from sterile leaf and petiole explants and from in-situ-collected inflorescence buds of Petasites hybridus was achieved by a simple two-step protocol. Murashige and Skoog (MS) nutrient medium was supplemented with 17.6 μm benzyladenine (BA)+0.54 μm naphthaleneacetic acid (NAA) to induce shoots. After 5 weeks of culture, 40% of the petiole and 27% of the leaf explants
produced shoots compared to 76% of the inflorescence buds. Single shoots were excised and subcultured on MS medium supplemented
with various cytokinins (N6-(Δ2-isopentenyl)adenine, BA, kinetin and thidiazuron). A concentration of 8.8 μm kinetin+0.54 μm NAA performed best in terms of shoot multiplication rate, average shoot length and spontaneous root induction.
Received: 20 August 1997 / Revision received: 29 December 1997 / Accepted: 5 February 1998 相似文献
2.
An in vitro method for obtaining plants of Acacia catechu has been developed using nodal explants from mature `elite' trees growing in the field. Maximum shoot bud development (eight
to ten) from a single explant was achieved on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP)
(4.0 mg/l) and α-naphthaleneacetic acid (0.5 mg/l). Addition of adenine sulphate (25.0 mg/l), ascorbic acid (20.0 mg/l) and glutamine (150.0
mg/l) to the medium was found beneficial for maximum shoot bud induction. The shoot buds developed into healthy and sturdy
shoots on MS medium containing BAP and kinetin at 1.0 mg/l. Excised shoots were rooted on 1/4-strength MS medium with indole-3-acetic
acid at 3.0 mg/l and 1.5% sucrose to obtain complete plants.
Received: 17 June 1997 / Revision received: 11 September 1997 / Accepted: 27 September 1997 相似文献
3.
Leaf explants of Jatropha curcas cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ; 0.90 μM) in combination with indole-3-butyric
acid (IBA; 0.98μM) produced adventitious shoot buds directly on the surface of the explants without formation of intervening
callus while shoot bud formation was accompanied with callus formation on medium supplemented with 6-benzylaminopurine (BAP;
13.3 μM) and IBA (2.46 μM). TDZ treatment resulted in more than twice higher rate of shoot bud induction than BAP. Shoot buds
were multiplied and elongated following repeated transfers to medium containing BAP (2.22 μM) and gibberellic acid (GA3; 1.44 μM). The effect of copper sulphate on differentiation of shoot buds from leaf segments was also investigated. Both
shoot induction and multiplication media were supplemented with different levels of CuSO4 (0–5 μM). Significant improvement in shoot bud induction was observed when the concentration of CuSO4 was increased to 10 times the normal MS level. Healthy elongated shoots were rooted on half strength MS medium supplemented
with IBA (2.46 μM). Rooted plantlets were transferred to field and survived. Histological analysis revealed direct formation
of shoot buds from leaf explants. 相似文献
4.
A simple, high-frequency and reproducible protocol for induction of adventitious shoot buds and plant regeneration from leaf-disc
cultures of Jatropha curcas L. has been developed. Adventitious shoot buds were induced from very young leaf explants of in vitro germinated seedlings
as well as mature field-grown plants cultured on Murashige and Skoog’s (MS) medium supplemented with thidiazuron (TDZ) (2.27 μM),
6-benzylaminopurine (BA) (2.22 μM) and indole-3-butyric acid (IBA) (0.49 μM). The presence of TDZ in the induction medium
has greater influence on the induction of adventitious shoot buds, whereas BA in the absence of TDZ promoted callus induction
rather than shoot buds. Induced shoot buds were multiplied and elongated into shoots following transfer to the MS medium supplemented
with BA (4.44 μM), kinetin (Kn) (2.33 μM), indole-3-acetic acid (IAA) (1.43 μM), and gibberellic acid (GA3) (0.72 μM). Well-developed shoots were rooted on MS medium supplemented with IBA (0.5 μM) after 30 days. Regenerated plants
after 2 months of acclimatization were successfully transferred to the field without visible morphological variation. This
protocol might find use in mass production of true-to-type plants and in production of transgenic plants through Agrobacterium/biolistic-mediated transformation. 相似文献
5.
The morphogenetic potential of node, internode and leaf explants of Brahmi [Bacopa monniera (L.) Wettst.] was investigated to develop reliable protocols for shoot regeneration and somatic embryogenesis. The explants
were excised from shoots raised from axillary buds of nodal explants cultured on Murashige and Skoog (MS) basal medium. Presence
of 6-benzylaminopurine (BA) or kinetin influenced the degree of callus formation, from which a large number of shoot buds
regenerated. Leaf explants gave the largest number of shoot buds followed by node and internode explants. BA was superior
to kinetin; BA at 1.5 – 2.0 mg/l appeared to be optimum for inducing the maximum number of shoot buds. MS + 0.1 mg/l BA +
0.2 mg/l indole-3-acetic acid was the most suitable for shoot elongation. Elongated shoots were rooted on full- or half-strength
MS medium with or without 0.5 – 1.0 mg/l indole-3-butyric acid or 0.5 – 1.0 mg/l α-naphthaleneacetic acid. The rooted plants were successfully established in soil. Calli derived from nodal explants cultured
on MS medium containing 0.5 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), when subcultured on MS medium containing 0.1 or 0.5
mg/l BA or 0.2 mg/l 2,4-D + 0.1 or 0.5 mg/l kinetin, developed somatic embryos. The somatic embryos germinated either on the
same media or on MS basal medium, and the resulting plantlets were successfully transplanted to soil.
Received: 25 September 1996 / Revision received: 23 October 1997 / Accepted: 12 November 1997 相似文献
6.
Summary A method for adventitious shoot induction from petiole explants of Heracleum candicans is reported. Shoot buds were induced on Murashige and Skoog (MS) medium with 4.4μM 6-benzylaminopurine (BA) and 1.1 μM 2,4-dichlorophen-oxyacetic acid (2,4-D). A wound response in the presence of BA and 2,4-D at the time of culture was necessary
for inducing shoot buds. The shoot bud regeneration was significantly influenced by size, type and orientation of explants
on the culture medium. These shoot buds developed into 4–5 cm shoots upon transfer to a medium containing 1.1μM BA and 0.5 μM α-naphthaleneacetic acid (NAA). The regenerated shoots formed rooted plantlets on MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA). About 15 plants were established in the field for further evaluation. 相似文献
7.
In vitro plant regeneration of fig (Ficus carica L. cv. gular) using apical buds from mature trees 总被引:3,自引:0,他引:3
A reliable procedure for multiple-shoot induction and plantlet regeneration was developed with apical buds collected from
7- to 8-year-old trees of Ficus carica L. using Murashige and Skoog's (MS) medium supplemented with 2.0 mg/l 6-benzylaminopurine and 0.2 mg/l 1-naphthaleneacetic
acid. The in-vitro-regenerated shoots were further multiplied on MS medium supplemented with 2.0 mg/l 6-benzylaminopurine
and 0.2 mg/l 1-naphthaleneacetic acid and an average multiplication rate of four per subculture was established with 90% success.
Excised shoots were rooted in liquid half strength MS medium supplemented with 2.0 mg/l indole-3-butyric acid and 0.2% activated
charcoal. Regenerated plantlets were successfully established in soil, with a success rate of 68%.
Received: 28 July 1997 / Revision received: 15 January 1998 / Accepted: 7 February 1998 相似文献
8.
Leaf, stem, hypocotyl, cotyledon, root, shoot tip and embryo explants of Capsicum annuum L. cv. mathania were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (BAP) or kinetin (Kin) alone or in
combination with 3-indoleacetic acid (IAA), 3-indolebutyric acid (IBA), α-naphthaleneacetic acid (NAA) or 2,4-dichlorophenoxyacetic
acid (2,4-D). BAP (5.0 mgl−1) in the medium was found to be the best growth regulator for shoot bud differentiation. Shoot buds cultured on 5.0 mgl−1 BAP increased in number but did not elongate. For obtaining complete plantlets, shoot buds were placed on a medium with IBA
or NAA (0.1 mgl−1). Histological evidence revealed direct differentiation of buds from cotyledons. Regenerated plants were normal diploids.
Unorganized callus could not be induced to differentiate shoot buds. 相似文献
9.
Induction of direct somatic organogenesis in onion (Allium cepa L.) using a two-step flower or ovary culture 总被引:4,自引:0,他引:4
A novel method for direct organogenesis in onion (Allium cepa L.) resulting in the formation of multiple shoot structures induced on mature flower buds or ovaries in a two-step culture
procedure is described. Flowers were cultured on an induction medium containing 2 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D)
and 2 mg/l 6-benzylaminopurine (BAP). After 6 days (superior to 3 or 12 days), flowers or extracted ovaries were transferred
to a differentiation medium containing 2 mg/l thidiazuron (TDZ). Medium solidification with gellan gum was superior to agar
or agar/gellan gum mixture. A similar regeneration frequency was achieved at high (100 g/l) and lower (50 g/l) sucrose content.
Regeneration was obtained from all 12 cultivars or inbred lines examined, although the efficiency and the occurrence of hyperhydricity
varied depending on genotype and procedure used. Studies of plant growth regulators revealed that in the induction medium,
the auxin 2,4-D was superior to 5 mg/l naphthaleneacetic acid or picloram, which partially or completely inhibited regeneration.
Omitting cytokinin in the induction medium or substitution of BAP with 2 mg/l 2iP lowered regeneration, while substitution
with 1 mg/l TDZ was equally effective. In the differentiation medium, lower concentrations of TDZ (1 and 0.5 mg/l) or substitution
of TDZ with 5 mg/l BAP were equally or less effective.
Received: 14 October 1998 / Revision received: 19 November 1998 / Accepted: 30 November 1998 相似文献
10.
An efficient regeneration system for Phaseolus vulgaris was developed from mature seeds germinated on Murashige and Skoog (MS) medium supplemented with thidiazuron or N6-benzylaminopurine (BA) for 6 d. Using cotyledonary nodes, multiple buds were induced on the MS medium supplemented with 5.0
mg dm−3 BA with the induction frequency 71.9 % after 4-week culture. The buds were then transferred onto shoot formation medium containing
1.0 mg dm−3 BA, 0.1 mg dm−3 gibberellic acid and 2.0 mg dm−3 silver nitrate. The addition of AgNO3 enhanced the frequency of the shoot formation from 61.3 to 87.6 %. Root induction medium was half-strength MS medium with
0.75 mg dm−3 indolebutyric acid and 0.02 mg dm−3 BA. The average root frequency was 84.3 %. The regenerated plantlets with healthy roots grew successfully when transferred
to soil. Using this system we obtained over 10 regenerated plantlets from one explant. 相似文献
11.
Phenylacetic acid improves bud elongation and in vitro plant regeneration efficiency in Capsicum annuum L. 总被引:1,自引:0,他引:1
A highly efficient three-stage protocol for the regeneration of chilli pepper (Capsicum annuum L.) from cotyledon explants was developed. This protocol used PAA in both the shoot-bud induction medium and the medium for
elongation of the shoot buds. A superior medium for the induction of buds from the cotyledons was MS medium supplemented with
BA (5 or 7 mg/l) + PAA (2 mg/l). Buds were elongated during the second stage on medium containing BA (2 or 5 mg/l) + PAA (2 mg/l).
On this medium most of the buds elongated, and their number also increased due to the formation of new buds; bud elongation
was achieved in 100% of the cultures provided the buds were induced in the primary stage on a medium supplemented with BA+PAA.
The shoots that elongated in the second-stage rooted at 100% frequency on a medium supplemented with NAA (1 mg/l). The complete
plantlets with well-developed root and shoot systems were transferred to field conditions where they grew to maturity, flowered
and fruited normally. While shoot-bud induction from the cultured cotyledons was also observed on media supplemented with
BA (5 or 7 mg/l) alone or in combination with IAA (0.2–2 mg/l), buds induced on these media were often distorted, with most
not developing into normal shoots in the second-stage subculturing; a rosette of buds was seen in the second stage subculturing.
On the other hand, PAA in combination with BA in the primary induction medium and second-stage medium promoted normal development
and the elongation of shoot buds.
Received: 28 July 1998 / Revision received: 22 December 1998 / Accepted: 19 February 1999 相似文献
12.
Jaroslav Ďurkovič 《Trees - Structure and Function》2008,22(4):597-602
Sprouting axillary buds sampled from a mature 27-year-old shrub of Cornus mas ‘Macrocarpa’ were used as starting material for in vitro culture establishment. Multiple shoot cultures, grown on basal woody
plant medium with the pH adjusted to 5.6–5.7 and supplemented with 6-benzylaminopurine in combination with 1-naphthaleneacetic
acid, were capable of continuous axillary and adventitious shoot proliferation up to 1 year. Later on, growth ceased, shoot
tip necrosis appeared and shoot cultures died. Transfer of living shoots onto modified woody plant medium with the pH adjusted
to 6.8–7.0 led to vigorous growth of multiple shoot cultures without any loss of multiplication rates or decreased vitality
for several years. The use of 6-benzylaminopurine in combination with 1-naphthaleneacetic acid proved superior to the application
of thidiazuron which induced a frequent formation of short and fasciated shoots. 1-naphthaleneacetic acid promoted in vitro
adventitious rooting frequency up to 73.3%, whereas indole-3-butyric acid was not effective. Ex vitro acclimatized plants
did not show any visually detectable morphological variation. 相似文献
13.
Tiwari Vaibhav Tiwari Kavindra Nath Singh Brahma Deo 《Plant Cell, Tissue and Organ Culture》2001,66(1):9-16
A mass in vitro propagation system for Bacopa monniera (L.) Wettst. (Scrophulariaceae), a medicinally important plant, has been developed. A range of cytokinins have been investigated
for multiple shoot induction with node, internode and leaf explants. Of the four cytokinins (6-benzyladenine, thidiazuron,
kinetin and 2-isopentenyladenine) tested thidiazuron (6.8 μM) and 6-benzyladenine (8.9 μM) proved superior to other treatments. Optimum adventitious shoot buds induction occurred at 6.8 μM thidiazuron where an average of 93 shoot buds were produced in leaf explants after 7 weeks of incubation. However, subculture
of leaf explants on medium containing 2.2 μM benzyladenine yielded a higher number (129.1) of adventitious shoot buds by the end of third subculture. The percentage
shoot multiplication (100%) as well as the number of shoots per explant remained the high during the first 3 subculture cycles,
facilitating their simultaneous harvest for rooting. In vitro derived shoots were elongated on growth regulator-free MS medium and exhibited better rooting response on medium containing
4.9 μM IBA. After a hardening phase of 3 weeks, there was an almost 100% transplantation success in the field.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
14.
Multiplication by adventitious shoot regeneration from root explants was found to be the most suitable method for the propagation
of Swertia chirata. A two-step system consisting of an initial 3-week cultivation on modified Murashige and Skoog medium supplemented with 3 μM 6-benzylaminopurine (BAP), followed by another period of 3 weeks on hormone-free medium was used. After rooting and acclimatization
micropropagated plants could be successfully cultivated in Nepal.
Received: 28 July 1997 / Revision received: 10 February 1998 / Accepted: 16 February 1999 相似文献
15.
A method for regenerating pigeonpea [Cajanus cajan (L.) Millsp.] plants has been developed using distal cotyledonary segments of mature seeds as explants. A large number of
shoot buds were induced directly from explants of genotypes T-15-15 and GAUT-82-90 when cultured on six different basal media
fortified with 22.2 μm N6-benzylaminopurine, 2.3 μm kinetin, and 271 μm adenine sulfate. The shoot buds developed into shoots when they were subcultured on the same medium but with one-tenth concentrations
of cytokinins and adenine sulfate. The shoots elongated by subculturing first two to three times on Murashige and Skoog (MS)
basal medium supplemented with 2.22 μm N6-benzylaminopurine and 0.54 μm
α-naphthaleneacetic acid or on half-strength MS medium containing 2.89 μm gibberellic acid, and then once on the same medium without growth regulators. Elongated shoots were rooted with 80–85% efficiency
on MS medium with 4.92 μm indole-3-butyric acid and the plantlets were transferred for hardening. Plants survival in pots was 70–75%. This method may
be useful for improving the crop through genetic manipulations.
Received: 11 August 1997 / Revision received: 12 January 1998 / Accepted: 30 January 1998 相似文献
16.
R. V. Sreedhar L. Venkatachalam R. Thimmaraju N. Bhagyalakshmi M. S. Narayan G. A. Ravishankar 《Biologia Plantarum》2008,52(2):355-360
Shoot buds were induced directly on either side of midrib from adaxial surface of immature leaf explants in Stevia rebaudiana Bertoni five weeks after culturing in Murashige and Skoog’s nutrient medium supplemented with 8.88 μM of N
6-benzylaminopurine and kinetin ranging from 4.65 to 6.98 μM. Immature leaves of 0.6 to 1 cm were found to produce best response
(93 %) with a highest number of 4.93 shoot buds per explant. For elongation of regenerated shoot buds, MS medium supplemented
with 30 g dm−3 sucrose and indole-3-butyric acid (IBA) ranging from 4.92 to 7.38 μM were found most suitable. The medium was further modified
to suit bioreactor cultivation of regenerated shoots wherein the use of two-fold MS salts and 60 g dm−3 sucrose resulted in a high biomass yield of 50.68 g dm−3 (m/v) accounting for about 590 micro-cuttings in three weeks. Best rooting of micro-cuttings occurred in half strength MS
medium supplemented with IBA ranging from 4.92 to 7.38 μM, 15 g dm−3 sucrose and gelled with 0.8 % agar. Rooted plants were successfully established in substrate containing sand, Vermicompost and garden soil in equal proportions and grown in greenhouse. This is the first report on direct shoot regeneration from
Stevia leaves. 相似文献
17.
Direct shoot organogenesis and plant regeneration in safflower 总被引:1,自引:0,他引:1
Summary Adventitious shoot buds were induced directly on the adaxial surface of the cotyledons of eight safflower cultivars after
14 d of culture initiation on Murashige and Skoog's (MS) medium supplemented with various levels of 6-benzylaminopurine (BA).
Maximum shoot organogenesis of 54.4% with 10.2 shoots per responding cotyledon was obtained with 8.87 μM BA in the cv. S-144. Regenerated shoots were classified into three groups on the basis of their morphological features and
were found to be correlated with the levels of BA. The highest number of normal shoots was obtained from 2.2 μM BA treatment. The regenerated shoots of group I (normal shoots) were rooted on half-strength MS medium supplemented with
5.3 μM α-naphthaleneacetic acid, 3% sucrose and 0.8% bacto-agar. Rooted plantlets were successfully transferred to soil and appeared
morphologically normal. Histological studies revealed that shoot buds originated adventitiously from subepidermal cells. 相似文献
18.
In vitro propagation of northern red oak (Quercus rubra) shoots was successful from cotyledonary node explants excised from 8-wk-old in vitro grown seedlings. Initially, four shoots per explant were obtained on Murashige and Skoog (MS) medium supplemented with 4.4 μM
6-benzylaminopurine (BA), 0.45 μM thidiazuron (TDZ), and 500 mg l−1 casein hydrolysate (CH) with a regeneration frequency of 64.7% after 3 wk. Subculturing explants (after harvesting shoots)
to fresh treatment medium significantly increased shoot bud regeneration (16.6 buds per explant), but the buds failed to develop
into shoots. A higher percentage (73.3%) of the explants regenerated four shoots per explant on woody plant medium (WPM) supplemented
with 4.4 μM BA, 0.29 μM gibberellic acid (GA3), and 500 mg l−1 CH after 3 wk. Explants subcultured to fresh treatment medium after harvesting shoots significantly increased shoot regeneration
(16 shoots per explant). Shoot elongation was achieved (4 cm) when shoots were excised and cultured on WPM supplemented with
0.44 μM BA and 0.29 μM GA3. In vitro regenerated shoots were rooted on WPM supplemented with 4.9 μM indole-3-butyric acid. A higher percentage regeneration response
and shoot numbers per explant were recorded on WPM supplemented with BA and GA3, than on MS medium containing BA and TDZ. Lower concentrations of BA and GA3 were required for shoot elongation and prevention of shoot tip necrosis. Each cotyledonary node yielded approximately 20
shoots within 12 wk. Rooted plantlets were successfully acclimatized. 相似文献
19.
Rapid clonal multiplication of Aegle marmelos (L.) Corr. (Rutaceae), a medicinal tree, was achieved by enhanced axillary bud proliferation in young single-node segments
of a 25-year-old tree cultured in Murashige and Skoog (MS) nutrient medium. Bud break was dependent on cytokinin supply, but
the synergistic combination of 2.5 mg l–1 6-benzylaminopurine (BAP) and 1.0 mg l–1 indole-3-acetic acid (IAA) induced the formation of 12.1 shoots of up to 5.2 cm length in 48% of the explants after 7 weeks
of culture. Explants of in-vitro-grown shoots – node, whole leaf, shoot tip and internode – were subcultured in the presence
of 0.05–2.5 mg l–1 BAP to produce 11.3, 18.4, 5.3 and 3.2 shoots and shoot buds at a 100%, 70%, 95% and 40% rate respectively, in 7 weeks. Different
shoot nodes and leaves were equally regenerative and adventitious organogenesis in the latter was confined to cut petiolar
ends. Nodal explants responded most favourably at low BAP (0.05–0.1 mg l–1) and produced uniform (3.8–5.3 cm) shoots facilitating their simultaneous harvest for rooting. Repeated subculturing through
five cycles of nodes and leaves of shoot cultures enabled continuous production of healthy callus-free shoots without any
sign of decline. Shoot cuttings (3.0–5.2 cm) were best rooted in half-strength MS medium with 0.5 mg l–1 IAA (70%) or 10.0 mg l–1 indole-3-butyric acid (90%). Eighty-eight percent of the rooted plants were established in polybags after hardening.
Received: 4 April 1996 / Revision received: 8 September 1997 / Accepted: 20 September 1997 相似文献
20.
A reproducible in vitro regeneration system for Nepalese kutki (Picrorhiza scrophulariiflora Pennell) was developed from in vitro leaf derived callus. Induction of more than seven shoot buds per explant was achieved on Woody plant medium (WPM) supplemented
with 0.53 μM α-napthaleneacetic acid (NAA) and 0.23 μM kinetin (KIN). The shoots were elongated on WPM supplemented with 0.44
μM 6-benzylaminopurine (BAP) and rooted on WPM supplemented with 5.3 μM NAA within 2 weeks. The random amplified polymorphic
DNA (RAPD) analysis indicated genetic uniformity of the micropropagated plants with its donor plants. 相似文献