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1.
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
2.
In vitro regeneration of Acacia mangium via organogenesis 总被引:1,自引:0,他引:1
Plant regeneration of Acacia mangium was achieved through organogenesis in callus cultures. Calli were induced from five types of explants (embryo axes and cotyledons
of mature zygotic embryos as well as leaflets, petioles and stems of seedlings) of A. mangium on MS (Murashige and Skoog, 1962) basal medium containing 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 13.95 μM kinetin (KT). Green or green purple compact nodules containing clusters of meristematic centers were induced
in these calli after transfer to MS basal medium containing 1.14–22.75 μM thidiazuron (TDZ) and 1.43–2.86 μM indole-3-acetic acid (IAA). A combination of 4.55 μM TDZ and 1.43 μM IAA promoted the
highest percentage of calli to form nodules, in 8–11% of calli derived from cotyledons, embryo axes, leaflets or petiole and
in 4% of calli derived from stems. Twenty-two percent of the nodules formed adventitious shoots on MS basal medium containing
0.045 μM TDZ. Shoots were elongated on MS medium containing 0.045 μM TDZ supplemented with 7.22 μM gibberellic acid. The medium
containing 10.75 μM NAA and 2.33 μM KT promoted rooting of 10% of the elongated shoots. Plantlets grew up well in the green house.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
3.
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
4.
Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant 总被引:1,自引:0,他引:1
T. Dennis Thomas 《Acta Physiologiae Plantarum》2007,29(5):455-461
Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented
with various concentrations of BAP (2–8 μM) or TDZ (2–8 μM) alone or in combination with NAA (0.5 and 1.0 μM) produced low
shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15,
25 or 50 μM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After
pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 μM), TDZ (6 μM),
BAP (4 μM) + NAA (1.0 μM), TDZ (6 μM) + NAA (0.5 μM). Control explants were incubated directly on the medium without any pretreatment.
The pretreatment of explants with 15 μM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum
response was observed on MS medium supplemented with 6 μM TDZ. In this medium, 96 % cultures responded with an average number
of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots
per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ
alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present
investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides. 相似文献
5.
Bambusa balcooa is one of the most commercially important bamboo species. Regeneration of this species by sexual means is impossible because
no seeds are set after flowering. Vegetative propagation is hindered due to bulky propagules, low rooting ability of the culm
and branch cuttings, and seasonal specificity. This makes in vitro-based methods of regeneration important. This paper describes an efficient micropropagation protocol for multiplication of
B. balcooa from nodal explants. Nodal segments were surface sterilized with 0.1% mercuric chloride for 10 min, and cultured on Murashige
and Skoog (MS) medium supplemented with 4.4 μM 6-benzylaminopurine (BAP), 2.32 μM kinetin (Kn), and gelled with 0.2% w/v gelrite. Eighty-five percent of explants could be established in vitro with 90% of these achieving bud break. In vitro-formed shoots were successfully multiplied in MS liquid medium supplemented with 6.6 μM BAP, 2.32 μM Kn, 2.5% v/v coconut water, and 100 mg l−1
myo-inositol. Subculturing shoots every 3 wk yielded a consistent proliferation rate of 4.11-fold without decline in vigor. Shoot
clusters, containing 5 to 8 shoots, were rooted with 87.5% success in 1/2 MS supplemented with 5.71 μM indole-3-acetic acid
(IAA), 4.9 μM indole-3-butyric acid (IBA), and 5.37 μM naphthaleneacetic acid (NAA) within 3 wk. Plants regenerated in this
manner were acclimatized in the greenhouse and under a shade net with 88% success. 相似文献
6.
Yaser Hassan Dewir Nisha Singh Shakira Shaik Ashley Nicholas 《In vitro cellular & developmental biology. Plant》2010,46(1):41-46
The present study reports a simple protocol for indirect shoot organogenesis and plant regeneration of Sutherlandia using rachis and stem segments. Different concentrations (0.0–68.08 μmol l−1) of thidiazuron (TDZ) were used for callus induction and shoot organogenesis. The highest percentage of callus formation
(97.5%) and the highest percentage of explants forming shoots (88.8%) were obtained from rachis explants cultured onto Murashige
and Skoog (MS) medium (Murashige and Skoog, Physiol. Plant. 15:473–495, 1962) supplemented with 45.41 μmol l−1 TDZ. Scanning electron microscopy demonstrated the early development of adventitious shoots derived from callus cultures.
Shoot clusters were further developed and grown in MS hormone-free medium. The presence of l-canavanine was determined by thin-layer chromatography and confirmed after column fractionation using silica gel and nuclear
magnetic resonance spectroscopy. Individual shoots were rooted on different concentrations and combinations of MS salt strength
and IBA. Half-strength MS salt medium supplemented with 24.6 μmol l−1 IBA was optimal for root induction in which 78% of shoots were rooted. The in vitro plants were successfully acclimatized in a growth chamber with a 90% survival rate. 相似文献
7.
A simple, high frequency, and reproducible method for plant regeneration through direct organogenesis from cotyledonary leaf
explants of Jatropha curcas was developed using Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) or 6-benzyl
aminopurine (BAP). Medium containing TDZ has greater influence on regeneration as compared to BAP. The induced shoot buds
were transferred to MS medium containing 10 μM kinetin (Kn), 4.5 μM BAP, and 5.5 μM α-naphthaleneacetic acid (NAA) for shoot
proliferation. The proliferated shoots could be elongated on MS medium supplemented with different concentrations and combinations
of BAP, indole-3-acetic acid (IAA), NAA, and indole-3-butyric acid (IBA). MS medium with 2.25 μM BAP and 8.5 μM IAA was found
to be the best combination for shoot elongation. However, significant differences in plant regeneration and shoot elongation
were observed among the genotypes studied. Rooting was achieved when the basal cut end of elongated shoots were dipped in
half strength MS liquid medium containing different concentrations and combinations of IBA, IAA, and NAA for 4 days, followed
by transfer to growth regulators free half strength MS medium supplemented 0.25 mg l−1 activated charcoal. Elongated shoot treated with 15 μM IBA, 5.7 μM IAA, and 11 μM NAA resulted in highest percent rooting.
The rooted plants could be established in soil with more than 90% survival rate. The method developed may be useful in improvement
of J. curcas through genetic modification. 相似文献
8.
An efficient protocol was developed for micropropagation of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes (CNs) derived from 18-d-old axenic seedlings on Murashige and
Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.1–10 μM). The highest shoot regeneration frequency (90%) and maximum
number (15.2 ± 0.20) of shoots per explant was recorded on MS medium amended with 0.4 μM TDZ. Continuous presence of TDZ inhibited
shoot elongation. In the primary medium, TDZ-initiated cultures were transferred to the secondary medium supplemented with
another cytokinin, 6-benzyladenine (BA), for shoot growth and elongation. Maximum (90%) shoot elongation with an average shoot
length of 5.4 ± 0.06 cm was observed at 5 μM BA. To further enhance the number of shoots per explant, mother tissue was repeatedly
subcultured on fresh shoot induction medium after each harvest of newly formed shoots. Thus, by adopting this strategy, an
average of 44 shoots per explant could be obtained. About 65% of in vitro regenerated shoots produced a maximum number (4.4 ± 0.2) of roots per shoot by a two-step culture procedure employing pulse
treatment and subsequent transfer of treated shoots to a low concentration of 0.2 μM indole-3-butyric acid along with phloroglucinol
(3.96 μM). The in vitro-raised plantlets were successfully acclimatized first under culture room conditions, then to greenhouse with 70% survival
rate. 相似文献
9.
Seabuckthorn (Hippophae rhamnoides) is a multipurpose small tree with unique berries of high nutritional and pharmaceutical values. A clonally propagated plant
originating from a 20-year-old tree of H. r. rhamnoides × mongolica hybrid cultivar Julia and seedling offspring of this cultivar were investigated regarding induction of shoot organogenesis
in leaf explants and in roots of intact seedlings, and induction of direct somatic embryogenesis in explants from shoot tissue.
The highest percentage of leaf explants showing shoot organogenesis was achieved (juvenile explants, 65%; adult explants,
75%) when incubated in Murashige and Skoog (MS) medium supplemented with either 4.5 μM of the phenylurea cytokinin thidiazuron
(TDZ) or 2.25 μM TDZ plus 2.2 μM 6-benzyladenine (BA), for juvenile and adult explants, respectively, both supplemented with
0.53 μM α-naphthaleneacetic acid (NAA). Juvenile explants developed on average 18 shoots per explant in the MS medium supplemented
with 4.5 μM TDZ, a four fold increase over those incubated on the medium supplemented with 2.25 μM TDZ and 2.2 μM BA. Adult
leaf explants grown on medium containing 2.25 μM TDZ and 2.2 μM BA medium produced 12 shoots per explant, while those grown
on medium containing 4.5 μM TDZ produced 5 shoots per explant. Shoot organogenesis was observed in roots of intact seedlings
pre-cultured on plain medium lacking nutrients (PM) or woody plant medium (WPM) salts and then grown on WPM salts supplemented
with 4.4 μM BA, 0.29 μM gibberrelic acid (GA3), and 57.0 μM indoleacetic acid (IAA). The number of shoots formed on each seedling
root system was ten fold higher when the pre-culture was in WPM medium indicating a promoting effect of mineral nutrients
in the pre-culture medium. Somatic embryogenesis was induced in both juvenile and adult leaf explants in 65 and 78% of the
explants, respectively, in MS-based medium supplemented with 2.0 μM N-(2-Chloro-4-pyridyl)-N
1-phenylurea (CPPU), 0.53 μM NAA and varying concentrations of BA. There was an interaction effect between MS salt strength
and BA concentration. The most effective medium for inducing somatic embryogenesis in juvenile explants contained half strength
MS salts and 2.2 μM BA and full strength MS salts and 13.2 μM BA for adult explants. 相似文献
10.
Dormant buds from a mature tree of Populus tremula ‘Erecta’ were incubated on a Murashige and Skoog (MS) medium supplemented with 1.0 μM thidiazuron (TDZ). Induced shoots were
then proliferated on medium of MS or Woody Plant Medium (WPM), or Driver and Kuniyuki Walnut (DKW) supplemented with varying
levels of benzyladenine (BA). Overall, shoots grown on MS medium supplemented with 1.25–2.5 μM BA exhibited the highest frequency
of shoot proliferation (>95%) and more than 60% of responding explants produced more than five shoots per explant. Shoot organogenesis
was induced from both leaf and petiole explants incubated on WPM medium containing BA, or TDZ, or zeatin. Among the different
cytokinins tested, zeatin induced the highest frequency (average 72.1%) of shoot organogenesis. None of explants survived
on media containing no cytokinins within 6–8 weeks following culture. Overall, a higher frequency of shoot regeneration was
obtained from petioles than from leaf explants. The highest frequency of regeneration was achieved when petioles were incubated
on WPM containing 10–20 μM zeatin. Addition of naphthaleneacetic acid (NAA) did not have a significant effect on shoot regeneration
in all treatments. Shoot organogenesis was directly induced from petiole explants without intervening callus. Regenerated
shoots were easily rooted on all tested media supplemented with 0.5 μM NAA. Rooted plants were transferred to potting mix
and grown in the greenhouse. 相似文献
11.
A novel protocol for callus-mediated shoot regeneration was established for an important medicinal and ornamental plant native
to South China, Curcuma kwangsiensis, using shoot base sections excised from seedlings in vitro as explant sources. The frequency of callus formation reached
91% for explants cultured on MS medium containing 1.4 μM TDZ, 4.4 μM BA and 2.3 μM 2,4-D. 8.2 shoots per callus was achieved
on MS medium supplemented with 1.4 μM TDZ, 17.8 μM BA and 2.7 μM NAA. Single shoots transferred into MS medium free of plant
growth regulator rooted well. Regenerated plants acclimatized ex vitro at 100%, and grew vigorously under shaded greenhouse
conditions. 相似文献
12.
Pumpkin ash (Fraxinus profunda (Bush) Bush) is at risk for extirpation by an exotic insect, the emerald ash borer (EAB). Pumpkin ash is limited to wetland
areas of the Eastern United States, and has been listed as an endangered species because of EAB activity. Pumpkin ash provides
many benefits to the ecosystem, and its wood is used in the manufacturing industry. In vitro regeneration provides an integral
tool for the mass propagation and genetic transformation of pumpkin ash to combat EAB. Therefore, a plant regeneration protocol
was developed for pumpkin ash. Aseptically extracted hypocotyls formed adventitious shoots following 4 weeks on Murashige
and Skoog (MS) medium supplemented with 0–22.2 μM 6-benzyladenine (BA) and 0–6.8 μM thidiazuron (TDZ) then transferred for
an additional 4 weeks on MS medium with Gamborg B5 vitamins plus 0.2 g L−1 glycine (B5G) containing 6.7 μM BA, 1 μM indole-3-butryic acid (IBA), and 0.29 μM gibberellic acid (GA3). As adventitious shoots developed, these were transferred to a MSB5G medium with 13.3 μM BA, 1 μM IBA, and 0.29 μM GA3 for shoot elongation. Elongated shoots were successfully micropropagated using MSB5 medium with 10 μM BA and 10 μM TDZ. Adventitious
root formation was as high as 94% using woody plant medium supplemented with 4.9 μM IBA with shoots cultured for 10 days in
the dark followed by culture under a 16-h photoperiod. Acclimatization to the greenhouse was successful and normal plant growth
was observed. This protocol will provide a means for genetic transformation for EAB resistance and mass propagation for conservation. 相似文献
13.
Somatic embryogenesis and whole plant regeneration was achieved in callus cultures derived from immature zygotic embryos of
Prosopis laevigata (Humb. & Bonpl. ex Willd.) M.C. Johnst., recently identified as chromium (Cr), cadmium (Cd), lead (Pb) and nickel (Ni) accumulator. Embryogenic calli were induced on Murashige and Skoog (MS) medium added with a mixture of organic components plus N-6 benzyladenine
(BA) (6.62 μM) and 2,4-dichlorophenoxyacetic acid (2,4-D) (2.26 μM) or thidiazuron (4.54–9.08 μM) and indole-3-acetic acid
(1.42 μM). Embryogenic calli transferred onto half-strength MS medium without plant growth regulators developed globular
embryos, of which 20% matured when treated with 3.75% (w/v) polyethylene glycol (PEG), and of these 50% fully differentiated into plantlet embryo. Regenerated plants were successfully
acclimatized (90%), while in vitro seedlings transferred to MS medium containing 0.5 mM Cd, Cr, Ni or Pb, exhibited high heavy
metals accumulation (627 mg Cr kg−1, 5,688 mg Cd kg−1, 1,148 mg Ni kg−1, and 3,037 mg Pb kg−1 dry weight) and efficient roots to shoots translocation (42–73%). 相似文献
14.
Muthu Thiruvengadam K. T. Rekha Chang-Hsien Yang Narayanasamypillai Jayabalan Ill-Min Chung 《Plant biotechnology reports》2010,4(4):321-328
An efficient protocol for in vitro organogenesis was achieved from callus-derived immature and mature leaf explants of Momordica charantia, a very important vegetable and medicinal plant. Calluses were induced from immature leaf explants excised from in vitro
(15-day-old seedlings) mature leaf explants of vivo plants (45 days old). The explants were grown on Murashige and Skoog (MS)
medium with Gamborg (B5) vitamins containing 30 g l−1 sucrose, 2.2 g l−1 Gelrite, and 7.7 μM naphthalene acetic acid (NAA) with 2.2 μM thidiazuron (TDZ). Regeneration of adventitious shoots from
callus (30–40 shoots per explant) was achieved on MS medium containing 5.5 μM TDZ, 2.2 μM NAA, and 3.3 μM silver nitrate (AgNO3). The shoots (1.0 cm length) were excised from callus and elongated in MS medium fortified with 3.5 μM gibberellic acid (GA3). The elongated shoots were rooted in MS medium supplemented with 4.0 μM indole 3-butyric acid (IBA). Rooted plants were
acclimatized in the greenhouse and subsequently established in soil with a survival rate of 90%. This protocol yielded an
average of 40 plants per leaf explant with a culture period of 98 days. 相似文献
15.
Xingyu Yang Jinfeng Lü Jaime A. Teixeira da Silva Guohua Ma 《Plant Cell, Tissue and Organ Culture》2012,109(2):213-221
Primulina tabacum is a rare and endangered species that is endemic to China. Establishing an efficient regeneration system is necessary for
its conservation and reintroduction. In this study, when leaf explants collected from plants grown in four ecotypes in China
are incubated on Murashige and Skoog (MS) medium containing 5.0 μM thidiazuron (TDZ) for 30 days, then transferred to medium
containing 5.0 μM 6-benzyladenine (BA), adventitious shoots are then observed. Conversely, when leaf explants are incubated
on medium containing 5.0 μM BA for 30 days, then transferred to medium containing 5.0 μM TDZ, somatic embryogenesis is induced.
This indicates that somatic embryogenesis and shoot organogenesis could be switched simply by changing the order of two cytokinins
supplemented in the culture medium. Histological investigation has revealed that embryogenic cells are induced within 30 days
following incubation of explants in medium containing TDZ. Only if embryogenic cells were induced, TDZ could enhance somatic
embryogenesis and BA could stimulate shoot organogenesis. When comparing explants from different ecotypes, leaf explants
from Zixiadong in Hunan Province could induce low numbers (1–2) of either somatic embryos or adventitious shoots on medium
containing either 5.0 μM TDZ or 5.0 μM BA, respectively. Whereas, leaf explants from plants collected from the other three
ecological habitats could induce 50–70 somatic embryos/adventitious shoots per explant. Moreover, somatic embryos could induce
secondary somatic embryogenesis and adventitious shoots on different media. All regenerated shoots developed adventitious
roots when these are transferred to rooting medium, and over 95% of plantlets have survived following acclimatization and
transfer to a potting mixture (1:1, sand:vermiculite). 相似文献
16.
Murugesan Dhandapani Doo Hwan Kim Seung-Beom Hong 《In vitro cellular & developmental biology. Plant》2008,44(1):18-25
High-frequency plant regeneration of C. roseus cv. ‘little bright eye’ via somatic embryogenesis and organogenesis from five out of six explants was standardized. Two factors
were found to be important for regeneration: (1) the type of explants, and (2) the combination and concentrations of plant
growth regulators. The highest regeneration percentage through somatic embryogenesis was obtained from mature zygotic embryo
in MS medium supplemented with 7.5 μM of thidiazuron (TDZ). The mature embryo also regenerated efficiently via organogenesis
in MS medium supplemented with either 2.5 μM TDZ or 5.3 μM α-naphthalene acetic acid (NAA) and 2.2 μM 6-benzylaminopurine
(BA). Hypocotyl and cotyledon did not induce somatic embryogenesis and organogenesis in TDZ-containing medium but gave a maximum
percentage of shoots in MS medium supplemented with 5.3 μM NAA and 2.2 μM BA. Stem nodes and meristem tips showed better regeneration
via organogenesis in the medium supplemented with NAA and BA and in lower concentrations of TDZ. 相似文献
17.
A simple and rapid method for multiple shoot formation in vitro from immature embryo axis explants of Carica papaya L. cvs. Honey Dew, Washington and Co2 is described. Multiple shoot regeneration was achieved by culture of the explants on
modified Murashige and Skoog (MS) medium supplemented either with thidiazuron (TDZ; 0.45–22.7 μM) or a combination of benzylaminopurine
(BAP; 0.2 – 8.84 μM) and naphthalene acetic acid (NAA; 0.5 – 2.64 μM). Highest frequency of shoot regeneration occurred on
medium supplemented either with 2.25 μM TDZ or a combination of BAP (4.4 μM) and NAA (0.5 μM). Composition of the basal media
influenced the frequency of multiple shoot initiation. Stunted shoots regenerated at 4.5 μM and higher concentrations of TDZ.
Such shoots could, however, be elongated by transfer to medium containing 5.7 μM GA3. Rooting of the regenerated shoots was achieved in presence of indolebutyric acid (IBA; 4.92 – 19.68 μM), however, least
response was in presence of 14.7 μM IBA. Rooted plants were hardened and transferred to pots.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
18.
A protocol for plant regeneration from mesophyll and callus protoplasts of Robinia pseudoacacia L. was developed. For leaves from in vitro raised shoots, an enzyme combination of 2.0% cellulose and 0.3% macerozyme for
a digestion period of 20 h resulted in the best yield of protoplasts (9.45 × 105 protoplast/g fresh weight). Mesophyll-derived protoplasts started cell wall regeneration within 24 h of being embedded in
Nagata and Takebe (NT) medium supplemented with 5 μM NAA and 1 μM BAP followed by the first cell division on day three of
culture and micro-colony (32 cells) formation within day 7–10 in the same medium. However, using callus as the starting material,
a combination of 2.0% cellulose and 1.0% macerozyme for a digestion period of 24 h gave the highest protoplast yield (3.2 × 105 protoplast/g fresh weight). Cell wall regeneration in callus-derived protoplasts started within 24 h followed by the first
cell division on the day three (96 h) and the appearance of microcolonies of more than 32 cells by the end of first week (144 h)
of culture on solid WPM medium supplemented with 5 μM NAA and 1 μM BAP. Microcalli were visible to the naked eye after 45 days
on solid WPM medium. Proliferation of macro-calli was successfully accomplished on solid Murashige and Skoog (MS) medium with
5 μM NAA and 5 μM BAP. Both mesophyll and callus protoplast-derived calli produced shoots on MS medium with 0.5 μM NAA and
1 μM BAP within 25–30 days and multiplied on MS medium with 1.25 μM BAP. Excised microshoots were dipped in 1–2 ml of 2.0 μM
IBA for 24 h under dark aseptic conditions and transferred to double sterilized sand for rooting. The flasks containing sand
were inoculated with Rhizobium for in vitro nodulation. Forty-five plants transferred to pots in the glasshouse established well. 相似文献
19.
M. Arshad J. Silvestre G. Merlina C. Dumat E. Pinelli J. Kallerhoff 《Plant Cell, Tissue and Organ Culture》2012,108(2):315-322
Shoot organogenesis from mature leaf tissues of two scented Pelargonium capitatum cultivars, ‘Attar of Roses’ and ‘Atomic Snowflake’, grown in the greenhouse, were optimized in the presence of thidiazuron
(TDZ). The protocol involved preculture of leaf sections on basal Murashige and Skoog (MS) medium supplemented with 10 μM
TDZ, 4.4 μM of 6-benzyladenine (BA) and 5.4 μM α-naphtaleneacetic acid (NAA) for a period of 2 weeks and followed by subculture
of explants to a fresh medium containing 4.4 μM BA and 5.4 μM NAA. Frequency of regeneration reached approximately 93% for
both cultivars, with the induction of more than 100 shoots per explant. Regenerated plantlets were rooted on half-strength
MS medium supplemented with 4.4 mM sucrose and 8.6 μM of Indole-3-acetic acid (IAA). All regenerated shoots from both cultivars
developed roots when transferred to organic soil mix, acclimatized, and successfully transferred to greenhouse conditions.
When regenerated shoots were transferred to hydroponic conditions, frequency of survival was 76.2 and 61.9% for ‘Attar of
Roses’ and ‘Atomic Snowflake’, respectively. 相似文献
20.
Xiuli Shen Michael E. Kane Jianjun Chen 《In vitro cellular & developmental biology. Plant》2008,44(4):282-288
The capacity for indirect shoot organogenesis of leaf and root explants of four Dieffenbachia cultivars were examined on a modified Murashige and Skoog (MS; Physiol Plant 15:473–495, 1962) medium supplemented with different plant growth regulators in 112 combinations. Callus formation was only observed from
leaf explants on MS supplemented with 1–10 μM thidiazuron (TDZ) and 0.5–1.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D) regardless
of cultivars. The combination of 5 μM TDZ and 1 μM 2,4-D resulted in the greatest callus formation frequency among the four
cultivars tested. Significant differences in callus and shoot formation from leaf explants were also observed among cultivars.
Cultivars Camouflage, Camille, Octopus, and Star Bright produced green nodular, brown nodular, yellow friable, and green compact
calli with corresponding maximum callus formation frequencies of 96%, 62%, 54%, and 52%, respectively. A maximum of 6.7 shoots/callus
was observed in cv. Camouflage, followed by cvs. Camille and Star Bright at 3.7 and 3.5, respectively. Calli of cv. Octopus
displayed no capacity for shoot organogenesis. Regardless of cultivar, callus formation was not observed on root explants.
Regenerated shoots were successfully acclimatized in a shaded greenhouse condition with 100% survival. 相似文献