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1.
Factors affecting successful establishment in vitro, rapid proliferation and rooting of apricot cultivar ‘Bebecou’ were studied. Ethanol and NaOCl were applied in several combinations for disinfection; chilling, plant growth regulators BA, IAA and GA3, antibiotics, different culture vessels and systems of subculture were evaluated for the optimization of shoot proliferation and the auxins NAA and IBA were assessed for root induction. The highest number of new microshoots/explant (18.7) was obtained in a culture medium supplemented with 2.2 μM BA+0.57 μM IAA after 300 h of chilling. The effect of GA3 (11.4 μM) on shoot proliferation was positive in combination with 4.4 or 8.9 μM BA. Shoot length and productivity were highest at 2.2 μM BA+11.4 μM GA3+0.57 μM IAA and at 2.2 μM BA+0.57 μM IAA, respectively and decreased as cytokinin concentration increased. The antibiotic ‘Na-cefotaxime’ had a minimal impact on shoot growth when used at the lowest concentration (250 mg l−1). Subculture every 2 weeks in a medium supplemented with 2.2 μM BA and 0.57 μM IAA was more efficient for shoot induction than alternation of 20 days culture in a propagation medium supplemented with 2.2 μM BA and 10 days culture in an elongation medium supplemented with 1.1 μM BA and 5.71 μM IAA. The highest number of roots/shoot (8.1) was recorded at 19.6 μM IBA. 相似文献
2.
Protocols for in vitro propagation of non-toxic variety of J. curcas through axillary bud proliferation and direct adventitious shoot bud regeneration from leaf segments have been established.
Shoot bud proliferation from axillaries was assessed on an initial basal Murashige and Skoog (MS) salt medium supplemented
with different concentrations of benzyladenine (BA), kinetin and thidiazuron (TDZ) followed by subculture to medium with 4.4-8.9
μM BA. Regardless of the concentration of BA in the subculture medium, shoot multiplication rate was optimum (10–12.3) with
primary culture on medium supplemented with 2.3–4.5 μM TDZ. Efficient adventitious shoot regeneration from leaf tissues was
achieved with culture on medium with 8.9–44.4 μM BA + 4.9 μM indole-3-butyric acid (IBA) followed by transfer to medium supplemented
with 8.9 μM BA + 2.5 μM IBA. Similarity index between toxic Indian variety and the non-toxic variety based on 435 RAPD markers
was 96.3%. Crossing studies followed by phorbol ester quantitation revealed that outcrosses with toxic J. curcas do not affect the phorbol ester content of seeds borne on the non-toxic variety. 相似文献
3.
Kottackal Poulose Martin A. K. Pradeep Joseph Madassery 《Acta Physiologiae Plantarum》2011,33(4):1141-1148
Trichopus zeylanicus subsp. travancoricus (known as Arogyapacha), an endangered ethnomedicinal plant of the Western Ghats of South India, serves as the major source
of the commercial drug Jeevani. The present study established a long-term high frequency in vitro propagation protocol for Arogyapacha. Callus obtained from
the branch–petiole explants cultured on Murashige and Skoog (MS) medium with 4.5 μM 2,4-dichlorophenoxyacetic acid upon subculture
to medium with different concentrations of 6-benzyladenine (BA) either alone or in combination with an auxin favoured shoot
morphogenesis. Medium with 13.3 μM BA alone facilitated high frequency shoot bud (mean of 93.2) formation. Medium with lower
concentrations of BA (4.4, 6.6 and 8.8 μM) alone or in combination with lower concentration of α-naphthaleneacetic acid (NAA)
or indole-3-butyric acid (IBA) favoured better shoot growth than 13.3 μM BA containing medium, but with reduced number of
shoot buds. Subsequent cultures on medium with lower concentrations of BA and also on MS basal media facilitated shoot formation
as well as growth of shoots. The shoot regeneration potential showed no decline up to 5 years. Culture of the in vitro-derived
whole branch–leaf explants on MS basal medium developed shoots directly from the node. On medium with 19.6 μM IBA, the whole
branch–leaf explants induced nodular callus from the node, which developed shoots later. Subsequent cultures on medium with
BA exhibited high frequency shoot formation. The transfer of shoots after 10–15 days culture on half-strength MS medium containing
2.7 μM NAA to half-strength basal medium induced a mean of 11.3 roots. Field survival of plantlets relied on the soil mix:
a 1:4 ratio of sand and red-soil exhibited the highest plantlets survival (86.6%). RAPD profile of the source plant and plants
regenerated from calli after 4 years showed no polymorphism. The established plantlets with morpho-floral features similar
to that of the source plants flowered normally and set fruits. 相似文献
4.
S. Y. Park Y. W. Kim H. K. Moon H. N. Murthy Y. H. Choi H. M. Cho 《Plant Cell, Tissue and Organ Culture》2008,93(3):341-346
The effect of phytohormones on the breaking of dormancy of axillary buds in Salix pseudolasiogyne and their subsequent proliferation from nodal explants were examined. Nodal explants obtained from a 20–year-old S. pseudolasiogyne tree were cultured either on woody plant basal medium (WPM) or WPM supplemented with benzyladenine (BA, 2.2/4.4 μM), zeatin
(1.1/2.2 μM), gibberillic acid (GA3, 2.9 and 14.5 μM), and GA3 + BA (2.9 + 4.4 μM). Although axillary shoots developed in all the media, a higher percentage bud break occurred on BA supplemented
media. To corroborate the results, endogenous levels of cytokinins [Cks, N
6-isopentenyladenine (iP), zeatin riboside (t-ZR), dihydrozeatinriboside (DHZR)] and abscisic acid (ABA) were determined. On BA supplemented media, the levels of zeatin
type (Z-type) of Cks were higher than those of isopentenyladenine type of Ck in the explants, while the ABA level was low.
Axillary shoots did not grow well and became necrotic upon subculture to fresh basal WPM. In order to improve shoot growth,
they were subcultured twice at a 4-week interval on to WPM supplemented with BA (2.2/4.4 μM), GA3 (1.4 μM), or GA3 + BA (1.4 + 4.4/2.9 + 4.4 μM). Maximal shoot growth (93%) was achieved on WPM supplemented with 2.2 μM BA. Comparative analyses
of endogenous Cks revealed that higher Cks (Z-type Cks) were present in actively growing shoots. Rooting was readily achieved
when the shoots were subcultured to WPM without phytohormones. The rooted plants were acclimatized well upon transplantation. 相似文献
5.
A rapid and efficient micropropagation system was developed for Psoralea corylifolia, an endangered, valuable medicinal plant. Multiple shoot buds were obtained in half-strength liquid Phillips–Collins (L2)
medium supplemented with 5 μM benzylaminopurine (BA) and 5 μM thidiazuron (TDZ) from apical bud explants of 1-week-old cultures.
The shoot buds were subcultured on enriched solid L2 medium supplemented with different concentrations and combinations of
BA, kinetin (KIN), 2-isopentenyladenine (2iP), TDZ, bavistin (BVN) and trimethoprim (TMP). Enriched solid L2 medium supplemented
with 2 μM BA, 1 μM TDZ and 100 mg l−1 BVN were more effective in producing greater number of shoots per explant (85.2 ± 0.9 shoots/explant) after 4 weeks of culture.
The regenerated shoots (40–50 mm in length) rooted and accompanied by hardening upon transfer to 50 μM indole-3-butyric acid
(IBA) for 15 min and followed by planting in sterile soil mixture and vermiculate (3:1 v/v), with 50 ml of one-eight strength
L2 basal salt solution devoid of sucrose and inositol, supplemented with 5 μM IBA and 100 mg l−1 BVN. The plants achieved 100% rooting with hardening. Subsequently the rooted plants were successfully established in the
field. The survival percentage differed with seasonal variations. The concentration of psoralen was evaluated in different
tissues of ex vitro and in vivo grown plants by high-performance liquid chromatography (HPLC). Psoralen content was increased
in leaves (2.97%), roots (2.38%), stems (5.40%) and seeds (1.63%) of ex vitro plants than the in vivo plants. This system
facilitates for commercial and rapid propagation of P. corylifolia for conservation strategies and phytomedicine production. 相似文献
6.
Hannibal T. Musarurwa Johannes van Staden Nokwanda P. Makunga 《Plant Growth Regulation》2010,61(3):287-295
The aromatic medicinal plant Salvia stenophylla contains α-bisabolol, making this plant an important contributor to the aromatherapy and cosmetic industries in South Africa.
Due to its commercial importance, the cultivation of this plant using an in vitro system was considered. Firstly, seedlings
were raised in vitro after breaking dormancy with light, smoke-water or chemical scarification treatments. Germination improved
when seeds were smoke-treated or soaked in 70% (v/v) H2SO4. Vigorous plantlet regeneration was achieved when seedling explants were cultured on Murashige and Skoog (1962) medium with 5.7 μM IAA and 8.9 μM BA. The potential regeneration capacity for this protocol was estimated and over 1,000
plantlets can be produced from a single shoot (6.67 cm with 4–6 nodes) over a period of 3 months. Plants rooted easily regardless
of their growth medium. This was followed by their successful rapid establishment and normal growth out of culture (75%).
Finally, the volatile compounds in in vitro plants were compared to ex vitro plants via headspace solid phase microextraction
linked to gas chromatography–mass spectrometry. The chemical complexity of microplants was similar to wild plants with in
vitro plants continuing to produce α-bisabolol (21%) at high levels. 相似文献
7.
Gerbera jamesonii H. Bolus ex Hook (Family: Asteraceae) has been successfully acclimatized from temperate to subtropical North Indian plains
of Lucknow through in vitro propagation. Flower heads were collected from greenhouse, segmented into 4–16 pieces and cultured
in Murashige and Skoog’s medium (MS) (Physiol Plant 15:472–497, 1962) supplemented with 2.87 μM indole-3-acetic acid (IAA)
and 8.88 μM N6-benzyladenine (BA) for shoot regeneration. Shoots were subcultured on growth regulator free MS medium. Apical shoot meristems
from in vitro plantlets of gerbera were tested in MS medium with different combination of cytokinins [BA, kinetin, and thidiazuron
(TDZ)] alongwith 2.68 μM 1-naphthaleneacetic acid (NAA) for shoot multiplication. The optimum results were obtained with 8.88 μM
BA. Regenerated plants with well-established root system were transferred to pots containing soil and sand (1:1 v/v) and were
kept in humidity chamber with 80–90% relative humidity for 0, 5, 10, 15, 20, and 25 days before they were transferred to field
(during October, 2005 to February, 2006). Survival percentage was higher when regenerated plantlets were kept under humidity
chamber for 15 days. An attempt was made to obtain basic information on different biochemical changes during acclimatization
process of in vitro raised plantlets. Increased lipid peroxidation and high H2O2 content in early stages of acclimatization process reflected a similar process of oxidative stress. Our work suggests that
tissue-cultured plants develop antioxidant enzymatic protective system which determine the ability to survive in oxidative
stress and up regulation of these enzymes would help to reduce the built up of reactive oxygen species (ROS). 相似文献
8.
Pterocarpus marsupium (Bijasal) is a valuable multipurpose forest tree. The regeneration rate in natural habitat is low and the tree is overexploited.
An in vitro propagation protocol has been developed from nodal explants obtained from in vitro raised 18-day-old axenic seedlings.
The highest shoot regeneration frequency (85%), maximum number of multiple shoots (8.6) as well as length (4.8 cm) were induced
from nodal explants on Murashige and Skoog (MS) medium amended with 4.0 μM 6-benzyladenine (BA), 0.5 μM indole-3-acetic acid
(IAA) and 20 μM adenine sulphate (AdS). The percentage of shoot multiplication as well as the number of shoots per node remained
the same during the first two subculture, afterwards a decline was recorded. Rooting was best induced in microshoots excised
from proliferated shoot cultures on semisolid hormone-free half-strength MS medium, after a pulse (dip) treatment for 7 days
in half-strength MS liquid medium containing 100 μM indole-3-butyric acid (IBA) and 15.84 μM phloroglucinol (PG). The in vitro-raised
plantlets were potted and acclimatized under culture room conditions for 4 weeks before their transfer to a greenhouse, where
the established plants showed 75% survival. 相似文献
9.
This paper describes an efficient in vitro micropropagation of Artemisia vulgaris using shoot tip and nodal explants. Among the various growth regulators tested, MS medium and B5 vitamins supplemented with BA (4.44 μM) and KN (2.32 μM) combination was found to yield a better response than BA (4.44–13.32 μM)
or KN (0.46–13.92 μM) alone in the medium. BA and KN combinations produced a maximum of 23.3 shoots per explant with 99.8%
shooting frequency. Multiple shoots raised were elongated on MS medium containing 0.44 μM BA and 1.44 μM GA3. Rooting was highest (98.2%) on MS medium containing 8.56 μM IAA. Rooted plantlets were successfully transferred to plastic
cups containing autoclaved garden soil, farmyard soil and sand (2:1:1) for hardening. After 65 days, the plantlets were transferred
to Botanical Evaluation Garden and maintained. The survival rate of plantlets varied under acclimatization. Plants looked
healthy with no visually detectable phenotypic variations. This is the first report on plant regeneration via organogenesis
of A. vulgaris. 相似文献
10.
Phillip A. Wadl Adam J. Dattilo Lisa M. Vito Robert N. Trigiano 《Plant Cell, Tissue and Organ Culture》2011,106(3):513-516
Pityopsis ruthii is an endangered herbaceous perennial species from the United States. In vitro multiplication of this species can be valuable
for germplasm conservation. Flower receptacles of P. ruthii were cultured on Murashige and Skoog medium (MS) supplemented with 11.4 μM indole-3-acetic acid (IAA) in combination with
2.2, 4.4 or 8.8 μM 6-benzyladenine (BA). Shoots were visible within 14–28 days and three plants were successfully rooted on
MS medium supplemented with 5.7 μM IAA. A two tailed t-test for paired-variates revealed that shoot regeneration on MS medium
amended with 11.4 μM IAA and 2.2 μM BA was significantly higher (P < 0.05) than on other treatments. Leaf explants were also cultured on MS not supplemented with growth regulators or supplemented
with 11.4 μM IAA in combination with 0, 2.2, 4.4 or 8.8 μM BA. Shoots were visible within 21–35 days and one plant was successfully
rooted on MS medium supplemented with 5.4 μM NAA. Shoot regeneration on MS medium augmented with 11.4 μM IAA and 2.2 μM BA
was significantly higher (P < 0.05) than the other treatments according to analysis of variance (ANOVA) with a rank transformation. Hyperhydricity and
rooting of shoots was problematic for explants derived from flower receptacles and leaf tissue, but viable plants were regenerated
using both explants sources indicating the potential role for micropropagation in the ex situ conservation of the species. 相似文献
11.
Vinod Kumar Ashwani Sharma Bellur Chayapathy Narasimha Prasad Harishchandra Bhaskar Gururaj Parvatam Giridhar Gokare Aswathanarayana Ravishankar 《Acta Physiologiae Plantarum》2007,29(1):11-18
Direct shoot bud induction and plant regeneration was achieved in Capsicum frutescens var. KTOC. Aseptically grown seedling explants devoid of roots, apical meristem and cotyledons were inoculated in an inverted
position in medium comprising of Murashige and Skoog (Physiol Plant 15:472–497, 1962) basal medium supplemented with 2-(N-morpholine) ethanesulphonic acid buffer along with 2.28 μM indole-3-acetic acid, 10 μM silver nitrate and either of 13.31–89.77 μM
benzyl adenine (BA), 9.29–23.23 μM kinetin, 0.91–9.12 μM zeatin, 2.46–9.84 μM 2-isopentenyl adenine. Profuse shoot bud induction
was observed only in explants grown on a media supplemented with BA (26.63 μM) as a cytokinin source and 19.4 ± 4.2 shoot
buds per explant was obtained in inverted mode under continuous light. Incorporation of polyamine inhibitors in the culture
medium completely inhibited shoothoot bud induction. Incorporation of exogenous polyamines improved the induction of shoot
buds under 24 h photoperiod. These buds were elongated in MS medium containing 2.8 μM gibberellic acid. Transfer of these
shoots to hormone-free MS medium resulted in rooting and rooted plants were transferred to fields. This protocol can be efficiently
used for mass propagation and presumably also for regeneration of genetically transformed C. frutescens. 相似文献
12.
An efficient and rapid method for in vitro clonal propagation of Huernia hystrix was developed, resulting in shoot regeneration within 3 weeks of culture. This endangered medicinal and ornamental succulent
is in high demand. Multiple shoots were regenerated from stem explants (10 mm length) cultured on Murashige and Skoog (MS)
medium containing 3% sucrose and supplemented with a range of NAA (0.00–8.06 μM) and BA (4.44–22.19 μM) concentrations. A
100% shoot response with a multiplication rate of four shoots per explant was obtained on MS medium containing 5.37 μM NAA
and 22.19 μM BA. Callus produced at the base of the explant on the same medium showed root organogenic potential. The in vitro
regenerated shoots produced roots when transferred to half strength MS medium with or without auxin. The micropropagated plants
were easily acclimatized within 2 months under greenhouse conditions when potted in a soil and sand mixture (1:1; v/v) treated
with a fungicide (Benlate, 0.01%). More than 95% survival with no observable morphological variations was obtained. The developed
protocol provides a simple, cost-effective means for the conservation of endangered H. hystrix by clonal propagation within a short time. 相似文献
13.
An efficient, rapid and large scale propagation of a multipurpose herb, Ocimum basilicum through in vitro culture of nodal segments with axillary buds from mature plants has been accomplished. Among the cytokinins,
6-benzyladenine (BA), thidiazuron (TDZ), kinetin (Kin) and 2-isopentenyl adenine (2-iP) tested as supplements to Murashige
and Skoog (MS) medium, 5.0 μM BA was optimum in inducing bud break. The highest rate of shoot multiplication was achieved
on half-strength MS medium supplemented with 2.5 μM BA and 0.5 μM indole-3-acetic acid (IAA) combination. The shoots regenerated
from TDZ supplemented medium when subcultured to hormone-free MS medium considerably increased the rate of shoot multiplication
and shoot length by the end of third subculture. For rooting, MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA)
proved to be better than that supplemented with IAA or α-naphthalene acetic acid (NAA). The in vitro raised plantlets with
well developed shoots and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse
with 90% survival rate. Chlorophyll a and b, carotenoids and net photosynthetic rate were measured in leaves during ex vitro
acclimatization at 0, 7, 14, 21 and 28 days. Firstly these parameters showed a decreasing trend but subsequently increased
after 7 days of acclimatization. These findings indicate that the adaptation of micropropagated plants to ex vitro conditions
is more extended in time than generally accepted. 相似文献
14.
A protocol for in vitro propagation of Isodon wightii (Bentham) H. Hara from nodal segments was developed. Multiple shoots were successfully established on half strength MS medium
supplemented with 4.4 μM BA. Enhancement of shoot multiplication and elongation was achieved on half strength MS medium supplemented
with 4.4 μM BA and 1.4 μM GA3. The regenerated shoots were rooted successfully on half strength MS medium supplemented with 4.9 μM IBA. Acclimatization
of in vitro rooted shoots was successful. The in vitro regenerated plants grew well in the greenhouse without any phenotypic
changes. 相似文献
15.
Sylwia Zielińska Ewelina Pi?tczak Danuta Kalemba Adam Matkowski 《Plant Cell, Tissue and Organ Culture》2011,107(1):161-167
The composition of volatile organic compounds emitted by in vitro shoots of Agastache rugosa (Fischer & C.A. Meyer) O. Kuntze (Lamiaceae) was studied using headspace solid-phase microextraction–gas chromatography–mass spectrometry and compared to the those emitted
by adult plants and in vitro-germinated seedlings. Shoot-tip explants were cultured on a solid MS medium supplemented with
either 4.4 μM 6-benzyladenine (BA), 9.3 μM kinetin, or 0.45 μM thidiazuron and with either 0.57 μM indole-3-acetic acid (IAA)
or 0.41 μM picloram. Shoot proliferation was observed in all these treatments. The presence of these plant growth regulators
in the culture medium significantly influenced the composition of volatiles as well as morphogenetic responses observed. The
number and quality of regenerating shoots and frequency of axillary bud break were highest in medium containing the BA + IAA
combination. Sixty-five compounds were identified in the headspace of the in vitro-produced material and plants cultivated
in the field. The in vitro shoots emitted both hydrocarbon (limonene, α-pinene) and oxidized (menthone, isomenthone, pulegone)
monoterpenes. The composition of monoterpenes differed depending on the type of auxin—rather than cytokinin—in the medium.
The emission of phenylallyl compounds, such as estragole, a major compound in field-grown plants, was markedly lower in shoot
cultures. 相似文献
16.
Landi Sun Suiwen Hou Dali Wu Yingcong Zhang 《In vitro cellular & developmental biology. Plant》2008,44(5):396-400
This study describes a reliable protocol for callus induction and rapid mass propagation of the ecologically important plant,
Zygophyllum xanthoxylon (Bunge) Maxim. The optimum callus induction medium was Murashige and Skoog (MS) supplemented with 4.4 μM 6-benzylaminopurine
(BAP) and 2.7 μM α-naphthalene–acetic acid (NAA), on which the callus induction frequencies from different seedling explants
were all 100%. However, seedling-derived callus did not form regenerated shoots. In order to achieve shoot multiplication,
shoots were developed from cultured plumules, at an average of 3.1 shoots per explant, and the regenerated shoot tips were
further multiplied by subculture. The best shoot multiplication from shoot tips was achieved on MS supplemented with 5.4 μM
NAA and 22.2 μM BAP after 40 d of culture. Seventy-three percent of regenerated shoots formed roots when cultured on MS supplemented
with 8.6 μM IAA after 4 wk of culture. The plants that acclimatized successfully in sand flourished the following year, with
normal morphology and growth characteristics. 相似文献
17.
Nodular callus was induced at a high frequency on young purple red, 5–15 mm long laminae taken from in vitro grown plants of mangosteen. The optimal medium was composed of Murashige and Skoog (MS) nutrients supplemented with 2.22
μM benzyladenine (BA), 2.25 μM thidiazuron (TDZ), 500 mg l-1 polyvinylpyrrolidone (PVP 360 000) and 3% sucrose. A multiplication rate of two–three was obtained by subculture of the nodular
callus at 3–4-week intervals. Plantlet regeneration from the nodules was achieved by transfer to woody plant medium (WPM)
with 500 mg l-1 PVP, 0.4 μM BA and 3% sucrose and overlaying with half strength liquid MS containing 0.32 μM naphthaleneacetic acid (NAA),
0.13 μM BA and 3% sucrose. Elongated shoots were rooted to 100% when wounded at the base of shoot, dipped in 4.4 mM indolebutyric
acid (IBA) solution in the dark for 15 min and cultured on WPM supplemented with 1.11 μM BA, 0.25% activated charcoal, 34.5
μM phloroglucinol (PG) and 3% sucrose.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
18.
U. A. Jo H. N. Murthy E. J. Hahn K. Y. Paek 《In vitro cellular & developmental biology. Plant》2008,44(1):26-32
An efficient, simple micropropagation method was developed for Alocasia amazonica using corms in semisolid and liquid cultures. Explants were cultured onto Murashige and Skoog (MS) medium (Murashige and
Skoog, Physiol. Plant. 15:473–497, 1962) supplemented with different cytokinins (Benzyladenine [BA, 2.22–13.32 μM], kinetin [2.32–13.95 μM], Thidiazuron [TDZ, 0.45–4.54 μM])
and cytokinin in combination with auxins [naphthalene acetic acid (NAA, 0.54–5.37 μM)/indole acetic acid (IAA, 0.57–5.71 μM)/indole
butyric acid (IBA, 0.49–4.9 μM)]. All supplementary-induced shoot proliferation and the optimal results was on the medium
supplemented with 2.27 μM TDZ, which induced 5.1 shoots per explant. Among the different concentrations of sucrose (0–120 g
l−1) tested for shoot proliferation, 30 g l−1 was found suitable for corm cultures of Alocasia amazonica. The optimal shoot proliferation and biomass values were with the plantlets grown at 30 μmol m−2 s−1 photosynthetic photon flux (PPF) and 25°C. Liquid cultures found suitable for shoot proliferation and biomass accumulation
was compared to semisolid cultures. Comparative studies of bioreactor systems [continuous immersion (with or without net)
and temporary immersion in liquid media using ebb and flood] revealed that shoot multiplication and growth were greatest with
the raft bioreactor system. Plantlets (cormlets) from the bioreactor were hydroponically cultured for 30 days, and 100% of
plants were acclimatized successfully. The simple efficient method of production of plantlets (cormlets) is useful for large-scale
multiplication of this important ornamental plant.
An erratum to this article can be found at 相似文献
19.
An efficient regeneration protocol for rapid multiplication of Melia azedarach, an economically as well as medicinally important timber-yielding tree, was developed. Nearly 90% of the culture exhibited
axillary bud sprouting and multiple shoot formation from nodal segments derived from 20-year-old candidate plus tree on Murashige
and Skoog (MS) medium supplemented with 5 μM 6-benzyladenine (BA). The highest shoot regeneration frequency (92%), maximum
number of multiple shoots (19.7 ± 0.31) as well as shoot length (4.9 ± 0.08 cm) was induced from nodal explants on MS medium
amended with 5.0 μM BA, 0.5 μM indole-3-acetic acid (IAA) and 30 μM adenine sulfate (AdS). Addition of 250 mg l−1 ammonium sulphate, (NH4)2SO4, and 100 mg l−1 K2SO4, prevented defoliation and tip burning without affecting the number of shoots. The explant harvest period also influenced
the bud break and shoot sprouting from nodal segments. Repeated subculturing of nodal explants on fresh MS medium containing
lower concentration of BA (2.5 μM) along with IAA (0.5 μM), AdS (30 μM) and additives was found most suitable growth regulator
regime for achieving 1.2-fold increase in shoot multiplication rate. The percentage of shoot multiplication as well as the
number of shoots per node remained the same during first three subculture passages, afterwards a decline was recorded. About
90% of the in vitro regenerated shoots were successfully rooted ex vitro by giving a pulse treatment of 250 μM indole-3-butyric
acid for 15 min, followed by their transfer to thermocol cups containing soilrite. The raised plantlets were successfully
acclimatized first under culture room conditions, then to green house with 85% survival rate. 相似文献
20.
Kaitlin J. Palla Paula M. Pijut 《In vitro cellular & developmental biology. Plant》2011,47(2):250-256
A plant regeneration protocol was developed for white ash (Fraxinus americana L.). Hypocotyls and cotyledons excised from embryos were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine
(BA) plus thidiazuron (TDZ), and compared for organogenic potential. Sixty-six percent of hypocotyl segments and 10.4% of
cotyledon segments produced adventitious shoots, with a mean number of adventitious shoots per explant of 3.5 ± 0.9 and 2.5 ± 1.5,
respectively. The best regeneration medium (52% shoot formation; 47% shoot elongation) for hypocotyls was MS basal medium
containing 22.2 μM BA plus 0.5 μM TDZ, producing a mean of 3.9 ± 0.4 adventitious shoots. Adventitious shoots were established
as proliferating shoot cultures following transfer to MS medium with Gamborg B5 vitamins supplemented with 10 μM BA plus 10 μM
TDZ. For in vitro rooting, woody plant medium with indole-3-acetic acid (IAA) at 0, 2.9, 5.7, or 8.6 μM in combination with 4.9 μM indole-3-butyric
acid (IBA) was tested for a 5- or 10-d dark culture period, followed by culture under a 16-h photoperiod. The best rooting
(78% to 81%) of in vitro shoots was obtained with a 5 d dark culture treatment on medium containing 2.9 or 5.7 μM IAA plus 4.9 μM IBA, with an average
of 2.6 ± 0.4 roots per shoot. Rooted plants were successfully acclimatized to the greenhouse. This adventitious shoot regeneration
and rooting protocol will be used as the basis for experimental studies to produce transgenic white ash with resistance to
the emerald ash borer. 相似文献