共查询到20条相似文献,搜索用时 31 毫秒
1.
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
2.
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
3.
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
4.
Surya Prakash Johannes Van Staden 《In vitro cellular & developmental biology. Plant》2008,44(4):338-341
A method for in vitro regeneration of Searsia dentata from nodal and shoot tip explants derived from mature trees is outlined. Nodal explants produced multiple shoots from the
axis when cultured on Murashige and Skoog (MS) medium containing 3% sucrose supplemented with 0, 5, 7.5, 10, or 12.5 μM N
6-benzyladenine (BA). An average of 5.3 shoots was obtained from nodal explants on 10 μM BA. For shoot tip explants, however,
supplementation of α-naphthaleneacetic acid (NAA) with BA favored a caulogenic response. A maximum of 6.1 shoots were produced
per shoot tip explant on MS containing 7.5 μM BA plus 5.0 μM NAA. The in vitro-regenerated shoots produced roots when transferred
to full-strength MS medium containing 3% sucrose and 10 μM indole-3-butyric acid (IBA). The developed plantlets were transferred
initially to a mist house. After an initial acclimatization period of 3–4 mo, plantlets were shifted to the greenhouse where
they thrived for 9 mo. The standardized protocol for mass propagation of S. dentata should eliminate the dependence on natural stands of plants for traditional medicinal purposes, and will also serve as a
means of conservation as the species is heavily overexploited. 相似文献
5.
Claudia Simões Norma Albarello Cátia Henriques Callado Tatiana Carvalho de Castro Elisabeth Mansur 《Plant Cell, Tissue and Organ Culture》2009,98(1):79-86
Alternative methods for in vitro shoot culture of Cleome rosea, a Brazilian herbaceous species with ornamental value and medicinal potential, were evaluated. A protocol for rapid in vitro
multiplication of roots, a valuable source of medicinal compounds, was also developed. Stem explants were cultured in liquid
media (continuous immersion and paper bridge), while root explants were cultivated in continuous immersion and on solidified
media. The highest numbers of shoots, 20 ± 4.6 shoots/explant, were obtained from stem explants incubated in a continuous
immersion system in a liquid medium supplemented with 2.2 μM BA. Root explants cultivated in liquid media produced only hyperhydrous
adventitious shoots. However, these explants generated 5.8 ± 0.8 shoots/explant by indirect organogenesis when cultivated
on solidified medium supplemented with 2.2 μM BA. In addition, root multiplication was achieved in liquid medium in the presence
of α-naphthaleneacetic acid. Adventitious shoots developed on newly formed roots when inoculated on solidified medium supplemented
with 2.2 μM BA. Shoot microcuttings developed roots when transferred onto solidified MS medium without growth regulators.
Rooted microcuttings were efficiently acclimatized when transferred ex vitro. 相似文献
6.
When cotyledonary explants, excised from in vitro germinated seedlings, of pomegranate (Punica granatum L.) were incubated on solid Murashige and Skoog (1962) medium supplemented with 21 μM naptheleneacetic acid (NAA) and 9 μM 6-benzyladenine (BA), 80% of explants developed callus.
A high frequency of shoot organogensis was obtained when explants were incubated on MS medium supplemented with 8 μM BA, 6 μM
NAA, and 6 μM giberrellic acid (GA3). However, adding 24 μM silver nitrate (AgNO3) to this medium markedly enhanced shoot regeneration frequency (63%) and mean number of shoots per explant (11.26) and length
of shoots (2.22 cm). Highest frequency of in vitro rooting, mean number of roots/shoot (4.32), and mean root length (2.71 cm)
were obtained when regenerated shoots were transferred to half-strength MS medium supplemented with 0.02% activated charcoal.
Well-rooted plantlets were acclimatized, and then transferred to soil medium. Moreover, when zygotic embryos of P. granatum, excised from seeds collected at 16 weeks following full bloom, were incubated on MS medium containing 30 g l−1 sucrose, 15% coconut water, 21 μM NAA, and 9 μM BA, they developed the highest frequency of embryogenic callus, clumps with
globular embryos, and mean number of both globular and heart-shaped embryos per callus clump. Subjecting zygotic embryo explants
to six-week dark incubation period was essential for embryogenic callus induction, and these were subsequently transferred
to 16 h photoperiod for further growth and development of somatic embryos. Germination of somatic embryos was observed when
these were transferred to MS medium was supplemented with 60 g l−1 sucrose. 相似文献
7.
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
8.
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
9.
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
10.
Multiple shoot induction and callus regeneration in Sarcostemma brevistigma Wight & Arnott, a rare medicinal plant 总被引:1,自引:0,他引:1
An efficient micropropagation protocol based on multiple shoot induction and callus regeneration has been standardized in
Sarcostemma brevistigma, a rare medicinal plant. The nodal cuttings were cultured on MS medium supplemented with BA (0.5–8 μM) or Kn (0.5–8 μM) alone
or in combination with NAA (0.5–1.5 μM). Maximum multiple shoot induction was observed on MS medium supplemented with 4 μM
BA. On this medium, 100% cultures responded with an average number of 11.3 shoots per explant. However, the average shoot
length was limited to only 0.9 cm on this medium. The addition of 1 μM NAA along with 4 μM BA gave rise to an average number
of 10.9 shoots with an average shoot length of 1.8 cm. Luxuriantly growing callus was obtained on MS medium supplemented with
BA (5 μM) and 2,4-D (2 μM). The callus was subcultured on MS medium supplemented with BA (2–15 μM) or Kn (2–15 μM) alone or
in combination with NAA (0.5–2 μM) for shoot organogenesis. Optimum callus regeneration was obtained on MS medium supplemented
with 10 μM BA and 1 μM NAA. On this medium, 100% cultures responded with an average number of 13.4 shoots per culture. The
shoots obtained via multiple shoot induction and organogenesis were rooted on half-strength MS medium supplemented with NAA
(1–7 μM) or IBA (1–7 μM). IBA was better than NAA in terms of both the percentage of cultures that responded and the average
number of roots per explant. The rooted shoots were successfully transplanted to soil with 86% success. This standardized
protocol will help to conserve this rare medicinal plant. 相似文献
11.
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
12.
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
13.
The genus Dierama comprises plants with a potential to be developed as ornamentals. D. erectum seeds were decontaminated and germinated on 1/10th strength Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) media
without plant growth regulators or sucrose. In an experiment investigating the effects of 6-benzyladenine (BA), meta-Topolin (mT), kinetin (KIN) and zeatin (Z) with or without α-naphthaleneacetic acid (NAA), the highest shoot number per hypocotyl (4.20 ± 0.51)
was obtained from MS medium supplemented with 1.0 μM Z after 8 weeks. This was followed by a combination of 2.0 μM KIN and
2.0 μM NAA with 3.67 ± 0.81 shoots per explant. BA treatments produced 3.20 ± 0.22 shoots per hypocotyl explant when 2.0 μM
was combined with 1.0 μM NAA, while mT gave 3.09 ± 0.99 shoots per explant when 2.0 μM mT was combined with 2.0 μM NAA. Adventitious shoot regeneration was optimised when shoots were grown under a 16-h photoperiod
at 100 μmol m−2 s−1 on MS medium supplemented with 1.0 μM BA. This resulted in an average of 12.73 ± 1.03 shoots per hypocotyl explant. Various
concentrations of ancymidol, activated charcoal and sucrose did not promote in vitro corm formation of this species. Plants
rooted successfully after 8 weeks on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) and had an average root
number of 2.73 ± 0.40. After 2 months of acclimatisation, plants had formed corms. The largest corms (of diameter 0.45 ± 0.03 cm)
were produced in plants pre-treated with 0.5 μM IBA. The highest plant survival percentage of 73% was also associated with
this treatment. 相似文献
14.
Cultures of seven Daphne species: Daphne caucasica, D. cneorum, D. giraldii, D. retusa, D. jasminea, D. laureola and D. tangutica were established in vitro on MS/WPM based media. Five of the species responded best on MS-based media (D. tangutica, D. laureola, D. caucasica, D. retusa and D. giraldii), while the remaining two species performed best on WPM-based media (D. cneorum, and D. jasminea). Shoot proliferation was achieved from both apical and nodal explants. Shoots were sub-cultured from stock cultures, cut
into nodal explants 3–5 cm long and place vertically on basal media supplemented with different concentrations and combinations
of cytokinins and auxins. Individual species displayed different responses to the various cytokinins and auxins. Among species,
D. jasminea produced the greatest proliferation rate with an average of 7.84 + 0.6 shoots per explant on WPM supplemented with 2.32 μM
BA + 0.0045 μM TDZ + 0.054 μM NAA, while the best multiplication rate for the same species grown on the same media supplemented
with a single cytokinin (BA) and no auxin was 2.60 + 1.3 shoots per explant. Following multiplication, new shoots transferred
to the elongation trails and then 50–100 mm Shoots used for rooting experiments. Increased rooting efficiencies were observed
on in vitro-generated shoots with the two-layer medium or dipping methods over when PGRs were uniformly incorporated into
the medium. Maximum rooting frequencies (average) ranged from 59% in D. tangutica to 85% in D. jasminea. Following in vitro rooting, rooted shoots immersed in 0.01% solution of humates and planted into a standard horticultural
substrate composed and watered weekly with a solution containing half-strength MS salts. 相似文献
15.
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
16.
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
17.
Epicotyl, petiole, and cotyledon explants derived from 14-d-old seedlings of Albizia odoratissima were cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of either 6-benzylaminopurine
(BAP) solely or in combination with 0.5 μM naphthalene-3-acetic acid (NAA). The percentage of shoot regeneration and the number
of shoots regenerated varied significantly depending on the type of explants used, the concentration of plant growth regulators,
and the orientation of explants on the culture medium. The best response in terms of the percentage of shoot regeneration
was obtained from epicotyls cultured horizontally on MS medium supplemented with 5 μM BAP, whereas the highest number of shoots
per responding explant was recorded on medium containing 2.5 μM BAP and 0.5 μM NAA. Successful rooting was achieved by placing
the microshoots onto MS medium containing 25 μM indole-3-butyric acid (IBA) for 24 h first, then transferring to the same
medium without IBA. Of the various substrates tested, vermiculite was the best for plant acclimatization, as 75% of the plants
survived and became established. 相似文献
18.
In vitro regeneration of Parkia timoriana (DC.) Merr. has been achieved using cotyledonary node explants. The ability to produce multiple shoots has been evaluated
using semi-solid Murashige and Skoog (MS) basal medium and Gamborg’s B-5 basal medium supplemented with various concentrations
of α-naphthalene acetic acid (NAA) and 6-benzylaminopurine (BA) either in single or in combinations. The explants cultured
in MS medium supplemented with combinations of 2.7 μM NAA and 11 μM BA showed the maximum frequency of multiple shoots (96.66%)
formation and number of shoots per explants (6.60), respectively. For rooting, full and half strength MS medium supplemented
with various concentrations of indole-3-butyric acid (IBA) and NAA were studied and the highest number of root formation was
observed in full-strength MS supplemented with 9.8 μM IBA. Using Agrobacterium tumefaciens strain EHA105 pCAMBIA2301 various optimum conditions for efficient transformation were determined by recording the percentage
of GUS+ explants. Following the optimized conditions, the co-cultured explants were cultured on semi-solid shoot regeneration medium
containing MS medium + 2.7 μM NAA + 11 μM BA + 100 mg/l kanamycin + 500 mg/l cefotaxime. After 8 weeks of culture, the regenerated
shoots were rooted in rooting medium (RM) containing MS medium + 9.8 μM indole-3-butyric acid (IBA), 3% sucrose, 7.5 mg/l
kanamycin and 500 mg/l cefotaxime. Successful transformation was confirmed by histochemical GUS activity of the regenerated
shoots, nptII gene PCR analyses of the regenerated kanamycin resistant plantlets and Southern analysis of putative transgenic PCR+ plants. 相似文献
19.
Mehmet Nuri Nas Leyla Gokbunar Nevzat Sevgin Murat Aydemir Merve Dagli Zahide Susluoglu 《Plant Growth Regulation》2012,67(1):57-63
The effects of culture media and cytokinin types on micropropagation of mature Crataegus aronia L. were investigated. Using single-axillary bud explants, the growth of cultures on MS, WPM, DKW and NRM containing 4.44 μM
benzyladenine (BA) plus 0.05 μM indole-3-butyric acid (IBA), and on NRM containing thidiazuron, meta-Topolin (mT) or BA at
1.25, 2.5, 5.0 or 7.5 μM plus 0.05 μM IBA were compared. The culture medium had significant effects on shoot number and length.
In comparison with MS, DKW and WPM, shoot production was greater on NRM (5.7 shoots per explant). Shoot production on MS,
DKW and WPM (4.2, 4.2 and 4.1, respectively) were statistically similar to each other. Thidiazuron was detrimental to shoot
formation and caused formation of rosette shoots and/or large callus to form on explants. In the presence of mT, only some
of the explants developed into shoots. Benzyladenine was the only cytokinin that promoted both shoot proliferation and shoot
elongation. Higher shoot numbers were obtained at 5.0 and 7.5 μM BA compared to lower concentrations of BA. Over 80% of microshoots
rooted and rooted shoots were successfully acclimatized to ex vitro conditions. 相似文献
20.
Mehdi Bakhshaie Mesbah Babalar Masoud Mirmasoumi Ahmad Khalighi 《Plant Cell, Tissue and Organ Culture》2010,101(2):229-235
A somatic embryogenesis (SE) protocol was established for the regeneration of Lilium ledebourii (Baker) Boiss. whole plants using new vegetative bulblet microscales and transverse thin cell layers (tTCLs) of young bulblet
roots as the explant sources. Bulblets were induced from bulb scale explants cultured for at least 3 months in the dark on
Murashige and Skoog (MS) medium containing 3% sucrose, 0.8% agar, and different concentrations of α-naphthaleneacetic acid
(NAA), 6-benzyladenine (BA), and thidiazuron. Embryo-like structures were obtained from tTCL explants of 3-month-old bulblets
(excised from bulb scale explants) following culture on solid MS medium containing 3% sucrose and various concentrations of
NAA and BA for 3 months in the dark. Both the explant source and the type of plant growth regulators affected the differentiation
of somatic embryos. The highest percentage (65.55%) of embryogenesis was obtained from bulblet microscale tTCLs cultured on
solid MS medium containing 0.54 μM NAA and 0.44 μM BA. Plants with normal shoots and roots were obtained following a 3-month
culture of embryos on growth regulator-free MS medium at 25 ± 1°C under a 16/8-h light/dark photoperiod (light intensity 40 μmol m−2 s−1, cool-white fluorescent light). The plants were successfully acclimatized in the growth chamber. 相似文献