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1.
Guohua Ma Jaime A. Teixeira da Silva Guojiang Wu 《Journal of Plant Growth Regulation》2011,30(1):114-116
The induction of adventitious buds from apical shoot explants of Euphorbia tirucalli was studied. On average, 10.5 adventitious buds were efficiently induced in a ring on the segment from one apical explant
on MS (Murashige and Skoog) medium supplemented with 0.5 mg l−1 thidiazuron and 0.5 mg l−1 benzylaminopurine. The adventitious buds could develop into adventitious shoots during subsequent cultures on hormone-free
MS medium. For rooting, shoot clumps were cultured on half-strength MS medium containing 0.2 mg l−1 α-naphthaleneacetic acid or indole-3-butyric acid. All the rooted plants survived establishment in soil within 2 months. 相似文献
2.
Eline Kirk Mørk Karin Henriksen Henrik Brinch-Pedersen Kell Kristiansen Karen Koefoed Petersen 《Plant Cell, Tissue and Organ Culture》2012,108(3):501-512
A protocol for adventitious shoot formation in Symphyotrichum novi-belgii was developed after investigating the effects of cultivar and hormone combinations. A Murashige and Skoog medium with 1.0 mg l−1 6-benzyladenine induced adventitious shoot formation in 15 out of 19 cultivars. Addition of 0.1 mg l−1 indole-3-acetic acid or naphthaleneacetic acid increased the total number of shoots per explant, but not the number of shoots
longer than 1 cm. Addition of dichlorophenoxyacetic acid (2,4-D) promoted callus formation, but inhibited shoot elongation.
A transformation system for the two cultivars Victoria Fanny and Victoria Jane was developed by co-cultivation of leaf explants
with Agrobacterium tumefaciens. Three bacterial strains (LBA 4404, A281 and C58) all carrying the binary vector, p35S-GUS-INT, and harbouring the uidA gene coding for β-glucuronidase (GUS) were used. Regeneration of transgenic plants after co-cultivation with A281 was independent
of cultivar, and all explants produced callus followed by indirect shoot formation. In ‘Victoria Fanny’ shoots were formed
faster and without a callus phase after co-cultivation with LBA 4404 or C58. The highest number of potentially transformed
shoots was regenerated after co-cultivation of ‘Victoria Fanny’ leaf explants with LBA 4404. Integration of the transgenes
in the plant genome was confirmed using PCR and Southern blot hybridisation. To verify that the transgenes could be transferred
to offspring, crosses were conducted between three transgenic lines of ‘Victoria Fanny’ and two wild type pollen donors. It
was demonstrated that viable seeds were produced and that the uidA gene was inherited. 相似文献
3.
A. V. Raghu Kuzhiyumparambil Unnikrishnan S. P. Geetha Gerald Martin Indira Balachandran 《In vitro cellular & developmental biology. Plant》2011,47(4):506-515
Embelia ribes, an important vulnerable medicinal liana, was regenerated through organogenesis and embryogenesis using leaf explants. Leaf
explants produced organogenic calluses on MS medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzylaminopurine. Shoot regeneration was obtained from organogenic calluses on MS medium containing different concentrations
of thidiazuron (TDZ) and indole-3-acetic acid (IAA). The frequency of shoot bud organogenesis was highest (23.9 shoots/explant)
in MS medium containing 0.5 mg l−1 TDZ and 0.1 mg l−1 IAA. The best result for induction of embryogenic callus was noticed in the combination of 2.0 mg l−1 TDZ and 0.5 mg l−1 2,4-D. This callus, maintained in the same medium, showed the highest differentiation of embryos (56.5%) after 6 wk of culture.
Embryos were transferred to MS medium supplemented with different concentrations of TDZ, and this facilitated conversion of
embryos into plants. After 6 wk of subculture, MS medium with 0.05 mg l−1 TDZ favored the highest percentage (52.2%) embryo conversion. As per the present protocol, 52.2% of the embryos underwent
conversion, and a mean number of 29.5 shoots per culture was obtained. Shoots developed from both types of calluses were rooted
on half-strength MS basal medium supplemented with 1.0 mg l−1 indole-3-butyric acid. HPLC-UV assay demonstrated the highest embelin content (5.33% w/w) in the embryogenic callus cultures. Embelin was isolated from embryogenic callus and was identified using IR and 1H NMR studies. 相似文献
4.
Guang-Zhe Lin Xiao-Mei Zhao Soon-Kwan Hong Yu-Ji Lian 《Plant Cell, Tissue and Organ Culture》2011,106(1):93-103
We have developed a system for the in vitro regeneration of pasqueflowers (Pulsatilla koreana Nakai). The system was based on somatic embryogenesis and shoot organogenesis. Over a growth period of 6 weeks, multiple
shoots were initiated from leaf, petiole, and pedicel explants on Murashige and Skoog (MS) medium containing 0.5 mg l−1 indole-3-acetic acid (IAA) and zeatin (Zn), kinetin (Kin), or 6-benzyladenine (BA). We achieved 100% of adventitious shoot
induced when petiole and pedicel explants were cultured on MS, 0.5–2.0 mg l−1 Zn, and 0.5 mg l−1 IAA. Somatic embryos developed from the explants and generated shoots on MS medium containing 0.25 mg l−1 Zn and 0.5 mg l−1 IAA. Globular and heart-shaped stages of somatic embryos were observed. Histological studies have revealed the stages of
development of somatic embryos. For propagation and growth, the regenerated shoots from organogenic or embryogenic calluses
were transferred to MS medium containing either (1) 1.5 mg l−1 Zn and 0.05 mg l−1 IAA or (2) 1.0 mg l−1 BA and 0.05 mg l−1 IAA. After the length of the shoots reached 3 cm, the shoots initiated by organogenesis as well as those initiated by somatic
embryogenesis were transferred to the root induction medium. After 2 months of culture in half-strength MS with 1.5 mg l−1 α-naphthalene acetic acid (NAA), the rooting ratio was 93%. Finally, the rooted plantlets were acclimatized in a mixture
of mountain soil and perlite. 相似文献
5.
This study describes culture conditions for a plant regeneration system via a combined pathway of somatic embryogenesis and
organogenesis in root explant cultures of the commercial rose cultivar 'Charming'. Root explants formed white calluses at
a frequency of 30% after 6 weeks of culture on Schenk and Hildebrandt (SH) medium supplemented with 11 mg l−1 2,4-dichlorophenoxyacetic acid. After 6 weeks of transfer to SH medium without growth regulators, initial white calluses
gave rise to globular somatic embryos at a frequency of 2.8%, which were subsequently dedifferentiated to embryonic tissues.
Somatic embryos or embryonic tissues initially derived from root explants did not undergo development beyond cotyledonary
stage. To produce adventitious shoots, embryonic tissues were sliced and cultured on SH medium with 0.5 mg l−1 6-benzyladenine. After 4 weeks of culture, 28% of embryonic tissue explants formed adventitious shoots. Regenerated shoots
were rooted on half strength SH medium with 0.1 mg l−1 α-naphthalaneacetic acid and subsequently grown to maturity. Root-derived embryonic tissues were proliferated by subculture,
while retaining the capacity for shoot production for a few years. 相似文献
6.
Nisar Ahmad Hina Fazal Bilal Haider Abbasi Muhammad Rashid Tariq Mahmood Nighat Fatima 《Plant Cell, Tissue and Organ Culture》2010,102(1):129-134
The organogenic potential and antioxidant potential (1, 1-diphenyl-2-picrylhydrazyl-scavenging activity) of the medicinal
plant Piper nigrum L. (black pepper) were investigated. Callus induction and shoot regeneration were induced from leaf explants of potted plants
cultured on MS medium supplemented with different plant growth regulators. The best callogenic response was observed on explants
cultured for 30 days on MS medium supplemented with either 0.5 or 1.5 mg l−1 6-benzyladenine (BA) + 1.0 mg l−1 α-naphthaleneacetic acid. Subsequent transfer of the callogenic explants onto MS medium supplemented with 1.5 mg l−1 BA + 1.0 mg l−1 gibberellic acid (GA3) achieved 85% shoot organogenesis after 30 days of culture. The maximum number (7.2) of shoots/explant was recorded for explants
cultured in MS medium supplemented with 1.0 mg l−1 BA. Following the transfer of shoots to an elongation medium, the longest shoots (5.4 cm) were observed on MS medium supplemented
with 1.0 mg l−1 BA + 1.0 mg l−1 GA3. The elongated shoots were rooted on MS medium supplemented with different concentrations of indole butyric acid. An assay
of the antioxidant potential of the in vitro-grown tissues revealed that the antioxidant activity of the regenerated shoots
was significantly higher than that of callus and the regenerated plantlets. 相似文献
7.
Bilal Haider Abbasi Mubarak Ali Khan Tariq Mahmood Mushtaq Ahmad Muhammad Fayyaz Chaudhary Mir Ajab Khan 《Plant Cell, Tissue and Organ Culture》2010,101(3):371-376
The morphogenic potential and free-radical scavenging activity of the medicinal plant, Silybum marianum L. (milk thistle) were investigated. Callus development and shoot organogenesis were induced from leaf explants of wild-grown
plants incubated on media supplemented with different plant growth regulators (PGRs). The highest frequency of callus induction
was observed on explants incubated on Murashige and Skoog (MS) medium supplemented with 5.0 mg l−1 6-benzyladenine (BA) after 20 days of culture. Subsequent transfer of callogenic explants onto MS medium supplemented with
2.0 mg l−1 gibberellic acid (GA3) and 1.0 mg l−1 α-naphthaleneacetic acid (NAA) resulted in 25.5 ± 2.0 shoots per culture flask after 30 days following culture. Moreover,
when shoots were transferred to an elongation medium, the longest shoots were observed on MS medium supplemented with 0.5 mg l−1 BA and 1.0 mg l−1 NAA, and these shoots were rooted on a PGR-free MS basal medium. Assay of antioxidant activity of in vitro and in vivo grown
tissues revealed that significantly higher antioxidant activity was observed in callus than all other regenerated tissues
and wild-grown plants. 相似文献
8.
Junli Wang Jue Wang Kun Liu Xuan Xiao Weizhen Gong Yuan Lu Mingfei Liu Dongting Xu 《In vitro cellular & developmental biology. Plant》2010,46(5):445-450
An efficient micropropagation system for Hylotelephium tatarinowii (Maxim.) H. Ohba, a rare medicinal plant, has been developed. Callus induced from leaf explants placed onto Murashige and
Skoog (MS) medium with supplementation of plant growth regulators. When the concentration of 2,4-dicholorophenoxy acetic acid
was as high as 2.0 mg l−1 in combination with 0.5 mg l−1 6-benzylaminopurine (6-BAP), the callus induction rate reached 92.1%. Adventitious shoots were observed on callus exposed
to 1.0 mg l−1 6-BAP, with 81.5% frequency of shoot regeneration after 30 d. Flower buds appeared after subculture. Regenerated shoots could
flower normally in vitro. Up to 100% of the regenerated shoots formed complete plantlets on half-strength MS medium without any growth regulator, with
an average of 5.9 roots per shoot explant. Quantitative analysis of flavonoids and rutin showed that the phytochemical profile
of callus and regenerated plants was similar to that of wild plants. 相似文献
9.
Ehsan Ullah Khan Xing-Zheng Fu Ji-Hong Liu 《Plant Cell, Tissue and Organ Culture》2012,109(2):383-390
In this study, attempts were made to develop a protocol for regeneration of transgenic plants via Agrobacterium tumefaciens-mediated transformation of leaf segments from ‘Valencia’ sweet orange (Citrus sinensis L. Osbeck) using gfp (green fluorescence protein) as a vital marker. Sensitivity of the leaf segments regeneration to kanamycin was evaluated,
which showed that 50 mg l−1 was the best among the tested concentrations. In addition, factors affecting the frequency of transient gfp expression were optimized, including leaf age, Agrobacterium concentration, infection time, and co-cultivation period. Adventitious shoots regenerated on medium containing Murashige
and Tucker basal medium plus 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.5 mg l−1 6-benzyladenine (BA) and 0.5 mg l−1 kinetin (KT). The leaf segments from 3-month-old in vitro seedlings, Agrobacterium concentration at OD600 of 0.6, 10-min immersion, and co-cultivation for 3 days yielded the highest frequency of transient gfp expression, shoots regeneration response and transformation efficiency. By applying these optimized parameters we recovered
independent transformed plants at the transformation efficiency of 23.33% on selection medium (MT salts augmented with 0.5 mg l−1 BA, 0.5 mg l−1 KT, 0.1 mg l−1 NAA, 50 mg l−1 kanamycin and 250 mg l−1 cefotaxime). Expression of gfp in the leaf segments and regenerated shoots was confirmed using fluorescence microscope. Polymerase chain reaction (PCR)
analysis using gfp and nptII gene-specific primers further confirmed the integration of the transgene in the independent transgenic plants. The transformation
methodology described here may pave the way for generating transgenic plants using leaf segments as explants. 相似文献
10.
Liquidambar styraciflua L. has great potential not only as an ornamental, but also for its commercial importance in pulp and paper production or
biomass energy. This study was designed to evaluate the influence of plant growth regulators on adventitious shoot multiplication
from shoot tips, and in vitro adventitious rooting. The morphogenic capacity of intact leaves grown in vitro was also assayed
for adventitious shoot formation and aerial root development. The highest shoot multiplication rate of 5.9 shoots per explant
was achieved with 0.7 mg l−1 6-benzylaminopurine plus 0.01 mg l−1 indole-3-butyric acid. Thidiazuron, alone or in combination with 6-benzylaminopurine, did not significantly support higher
shoot multiplication rates. The organogenic ability of the in vitro grown leaves was significantly lower and slower in comparison
with shoot tips. Microshoots rooted readily after transfer to a half-strength woody plant medium supplemented with 0.5–0.7 mg l−1 1-naphthaleneacetic acid, and were then successfully acclimatised to an ex vitro environment. A novel pattern of adventitious
rooting was observed from the aerial parts of microshoots which were not in contact with the medium, including the parenchyma
cells of the leaf blades as well as stem nodes and internodes. The regenerated plants established in soil did not show any
detectable morphological variation. 相似文献
11.
A simple protocol for direct shoot organogenesis and plant regeneration in Lessertia frutescens using hypocotyl and cotyledon segments is reported. l-canavanine content in the derived shoots is also quantified. Media containing different concentrations and combinations of
the cytokinins kinetin (K) and benzyladenine (BA) were tested for shoot induction potential. The best shoot regeneration rate
(83%) was obtained from hypocotyl segments cultured in Murashige and Skoog (MS) medium supplemented with 1 mg l−1 K; these hypocotyls also produced the largest number of shoots per explant (3.5) and the longest shoots per explant (13.3
mm). The best shoot regeneration rate (46%) using cotyledons as explant material was obtained in MS medium supplemented with
1 mg l−1 K and 1 mg l−1 BA or with 5 mg l−1 K and 0.5 mg l−1 BA. The highest number of cotyledon-derived shoots (1.5) was obtained in MS medium containing 2 mg l−1 K and 0.5 mg l−1 BA, and the longest cotyledon-derived shoots (6.1 mm) were obtained in MS medium containing 1 mg l−1 K and 0.5 mg l−1 BA. Shoots derived from hypocotyls cultured on media containing 1 mg l−1 K contained the highest quantity of l-canavanine (1.42 mg g−1) relative to the control (0.52 mg g−1). Shoots derived from cotyledons cultured on media containing 2 mg l−1 K contained the highest quantity of l-canavanine (2.07 mg g−1) compared to the control. Scanning electron microscopy revealed that shoots regenerated directly from the wounded epidermal
tissue, although callus formation was observed in most cultures. Young shoot clusters proliferated into healthy adventitious
shoots that were subsequently transferred directly onto rooting medium (MS medium containing 4 mg l−1 indole-3-butyric acid), eliminating the need for an additional multiplication or elongation phase. The in vitro plants were
successfully acclimatized in a growth chamber, achieving an 85% survival rate. 相似文献
12.
A. M. Vieitez E. Corredoira A. Ballester F. Muñoz J. Durán M. Ibarra 《Plant Cell, Tissue and Organ Culture》2009,98(2):135-145
North American oak species, with their characteristic strong episodic seasonal shoot growth, are highly problematic for clonal
micropropagation, resulting in the inability to achieve a stabilized shoot multiplication stage. The potential for initiating
and proliferating shoot cultures derived from Quercus alba, Q. bicolor and Q. rubra explants was investigated, and a micropropagation method for these species was developed. Branch segments from 6 to 7-year-old
trees were forced-flushed and the forced shoots were used as source of explants for culture initiation. A consistent shoot
multiplication stage was achieved, in 13 of the 15 genotypes established in vitro, although marked differences occurred in
explants from different genotypes/species. The control of efficient shoot multiplication involved the culture of decapitated
shoots in a stressful horizontal position on cytokinin-containing medium with a sequence of transfers within a 6-week subculture
cycle, which was beneficial to overcoming the episodic character of shoot growth. During each subculture cycle, the horizontally
placed explants were cultured on media containing 0.2 mg l−1 benzyladenine (BA) for 2 weeks with two successive transfers (2 weeks each) to fresh medium with 0.1 mg l−1 BA, giving a 6-week subculture cycle. The general appearance and vigor of Q. alba and Q. bicolor shoot cultures were improved by the inclusion of both 0.1 mg l−1 BA and 0.5 mg l−1 zeatin in the medium used for the second transfer within the 6-week subculture cycle. Addition of AgNO3 (3 mg l−1) to the shoot proliferation medium of Q. rubra had a significant positive effect on shoot development pattern by reducing deleterious symptoms, including shoot tip necrosis
and early senescence of leaves. The three species showed acceptable in vitro rooting rates by culturing microcuttings in medium
containing 25 mg l−1 indolebutyric acid for 48 h with subsequent transfer to auxin-free medium supplemented with 0.4% activated charcoal. Although
an initial 5-day dark period generally improved the rooting response, it was detrimental to the quality of regenerated plantlets.
However, activated charcoal stimulated not only the rooting frequencies, but it also enhanced plant quality, as evidenced
by root, shoot and leaf growth. 相似文献
13.
N. Praveen P. M. Naik S. H. Manohar A. Nayeem H. N. Murthy 《Acta Physiologiae Plantarum》2009,31(4):723-728
The major objectives of this study were to investigate an efficient rapid protocol for mass propagation of adventitious shoots
of brahmi using semisolid and liquid cultures; and to assess the amount of bacoside A accumulated in the regenerated shoots.
Leaf explants were grown in vitro on Murashige and Skoog semisolid medium supplemented with 0.5, 1.0, 1.5, 2.0 and 2.5 mg l−1 6-benzyladenine or kinetin (KN) or thidiazuron (TDZ) for 4 weeks. Adventitious shoots developed from leaf explants on all
cytokinin supplemented media. After 4 weeks of incubation, leaf explants were split into two batches and one set was subcultured
on semisolid medium and another set in liquid medium containing same concentration of cytokinins where they have come from.
Highest rate of shoot regeneration was observed for explants cultured on medium with 2 mg l−1 KN. The fresh and dry weight of shoots was also highest with this treatment. Liquid cultures were found suitable for proliferation
of shoots (155.6 shoots per explant) and they also favored highest biomass accumulation (8.60 g fresh and 0.35 g dry biomass).
The bacoside A contents were determined in shoots using HPLC. Analysis revealed that, the contents were highest with shoots
regenerated on medium supplemented with 2 mg l−1 KN. The amount of bacoside A was highest in the shoots regenerated in liquid medium (11.92 mg g−1 DW) and it was 2.2-fold higher compared to shoots grown on semisolid cultures. 相似文献
14.
Shi He-Ping Long Yong-Yue Sun Tie-Shan Tsang Po Keung Eric 《Plant Cell, Tissue and Organ Culture》2011,107(2):251-260
An efficient transformation system for the medicinal and aromatic plant, Pogostemon cablin Benth was developed by using agropine-type Agrobacterium rhizogenes ATCC15834. Hairy roots formed directly from the cut edges of leaf explants or via callus stage 8 days after inoculation with
the bacterium. The highest frequency of leaf explant transformation by Agrobacterium rhizogenes ATCC15834 was about 80% after infection for 25 days. Hairy roots grew rapidly on plant growth regulators (PGRs)-free Murashige
and Skoog (MS) or 6,7-V medium and had characteristics of transformed roots such as fast growth and high lateral branching.
The PCR amplification showed that rol genes of Ri plasmid of A. rhizogenes were integrated and expressed into the genome of transformed hairy roots. The hairy root line, PL6, grew very slowly in the
first 8 days, then grew very quickly between day 8 and day 24. The optimum medium for callus induction of hairy roots consisted
of 2.0 mg l−1 benzyladenine (BA) and 0.1 mg/l α-naphthaleneacetic acid (NAA); while optimum medium for adventitious shoot regeneration
from these cultures consisted of 0.1 mg l−1 BA and 0.1 mg l−1 NAA. Adventitious shoots could be rooted on 1/2MS. Southern blot analysis confirmed that rol genes of TL-DNA of Ri plasmid was integrated with at least three copies into the genome of hairy roots- regenerated P. cablin plants. The results presented provide a solid foundation for production of patchouli essential oil from hairy roots or its
regenerated plants and also provide possibilities for utilization of artifical polyploidization or chemical mutation of hairy
roots for improving germplasm and breeding of a new cultivar of P. cablin. 相似文献
15.
Su-Juan Zhao Zhong-Chun Zhang Xiang Gao Gulsum Tohsun Bao-Sheng Qiu 《Plant Cell, Tissue and Organ Culture》2009,99(1):9-16
An efficient micropropagation system for mining ecotype Sedum alfredii Hance, a newly identified Zn/Cd hyperaccumulator, was developed. Frequency of callus induction reached up to 70% from leaves
incubated on Murashige and Skoog (MS) medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxy acetic acid (2,4-D) and 0.5 mg l−1 6-benzyladenine (BA), and 83% from internodal stem segments grown on MS medium with 0.1 mg l−1 2,4-D and 0.1 mg l−1 BA. Callus proliferated rapidly on MS medium containing 0.2 mg l−1 2,4-D and 0.05 mg l−1 thidiazuron. The highest number of adventitious buds per callus (17.3) and frequency of shoot regeneration (93%) were obtained
when calli were grown on MS medium supplemented with 2.0 mg l−1 BA and 0.3 mg l−1 α-naphthalene acetic acid (NAA). Elongation of shoots was achieved when these were incubated on MS medium containing 3.0 mg l−1 gibberellic acid. Induction of roots was highest (21.4 roots per shoot) when shoots were transferred to MS medium containing
2.0 mg l−1 indole 3-butyric acid rather than either indole 3-acetic acid or NAA. When these in vitro plants were acclimatized and transferred
to the greenhouse, and grown in hydroponic solutions containing 200 μM cadmium (Cd), they exhibited high efficiency of Cd
transport, from roots to shoots, and hyperaccumulation of Cd. 相似文献
16.
An efficient plant regeneration system has been developed for figleaf gourd (Cucurbita ficifolia Bouché), which is exclusively used as a rootstock for cucumber. The protocol is based on results obtained from a series of
culture experiments involving different parts of the cotyledons and various media. The culture of cotyledon explants was critical
for the enhancement of shoot regeneration frequency. The lower parts of the cotyledon excised at the plumule base were found
to display a markedly enhanced production of adventitious shoots compared to other cotyledon regions. Culture in silver nitrate-supplemented
Murashige and Skoog (MS) medium was not beneficial for shoot regeneration and suppressed root regeneration. Efficient shoot
regeneration was obtained on MS medium containing 1.0 mg l−1 zeatin and 0.1 mg l−1 indole-3-acetic acid. Regenerated shoots successfully elongated and rooted in medium containing 0.1 mg l−1 1-naphthaleneacetic acid after 10–15 days of subculturing. The plantlets were satisfactorily acclimatized in a greenhouse
and grew into normal plants without any morphological alterations. 相似文献
17.
Meiru Li Hongqing Li Xiaoying Hu Xiaoping Pan Guojiang Wu 《Plant Cell, Tissue and Organ Culture》2011,106(3):363-371
Saussurea involucrata is a valuable traditional Chinese medicinal herb. This is the first report of a successful genetic transformation protocol
for S. involucrata using Agrobacterium tumefaciens. Leaf explants were incubated with A. tumefaciens strain EHA105 harboring the binary vector pCAMBIA 1301, which contains the hpt gene as a selectable marker for hygromycin resistance and an intron-containing β-glucuronidase gene as a reporter gene. Following
co-cultivation, about 23.7% of the explants produced hygromycin-resistant calli on MS basal medium (Murashige and Skoog in
Physiol Plant 15: 473–497, 1962) supplemented with 1 mg l−1 benzyladenine (BA), 0.1 mg l−1 α-naphthaleneacetic acid (NAA), 0.1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D), 20 mg l−1 hygromycin, and 500 mg l−1 cefotaxime. Shoots were regenerated following transfer of the resistant calli to shoot induction medium containing 1.5 mg l−1 BA, 0.1 mg l−1 NAA, 0.25 mg l−1 gibberellic acid (GA3), 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime, and about 67.5% of the resistant calli differentiated into shoots. Finally, 80% of the hygromycin-resistant shoots
rooted on MS media supplemented with 0.2 mg l−1 NAA, 20 mg l−1 hygromycin, and 250 mg l−1 cefotaxime. The transgenic nature of the transformants was demonstrated by detection of β-glucuronidase activity in the primary
transformants and by Southern blot hybridization analysis. About 16% of the total inoculated leaf explants produced transgenic
plants after approximately 5 months. Using this optimized transformation system, a rice ortholog of the Arabidopsis FLOWERING LOCUS T gene, Hd3a, was transferred into S. involucrata. Introduction of this gene caused an early-flowering phenotype in S. involucrata. 相似文献
18.
Bimal Kumar Ghimire Chang Yeon Yu Ill-Min Chung 《Plant Cell, Tissue and Organ Culture》2012,108(3):455-464
A simple and efficient procedure was developed for in vitro propagation of Solanum aculeatissimum Jacq. using leaf and petiole explants cultured on Murashige and Skoog (MS) medium supplemented with α-naphthalene acetic
acid (NAA) and 6-benzyladenine (BA). Effects of various plant growth regulators, explant types, carbohydrates, and basal salts
on induction of adventitious shoots were also studied. Leaf explants appeared to have better regeneration capacity than petiole
explants in the tested media. The highest regeneration frequency (79.33 ± 3.60%) and shoot number (11.33 ± 2.21 shoots per
explant) were obtained in leaf explants in MS medium containing 3% sucrose and 0.8% agar, supplemented with 0.1 mg/l NAA and
2.0 mg/l BA, whereas petiole explants were more responsive to 0.1 mg/l NAA and 1.0 mg/l thiadiazuron. Developed shoots rooted
best on MS medium with 1.0 mg/l indole acetic acid (IAA), producing 18.33 ± 2.51 roots per shoot. Histological investigation
showed that the shoot buds originated mainly from epidermal cells of wounded tissues, without callus formation. The regenerated
plantlets were successfully acclimatized in a greenhouse, where over 90% developed into morphologically normal and fertile
plants. Results of flow cytometry analysis on S. aculeatissimum indicated no variation in the ploidy levels of plants regenerated via direct shoot formation and showed almost the same phenotype
as that of mother plants. This adventitious shoot regeneration method may be used for large-scale shoot propagation and genetic
engineering studies of S. aculeatissimum. 相似文献
19.
Guirong Qiao Jing Zhou Jing Jiang Yuehua Sun Luanyin Pan Honggai Song Jingmin Jiang Renying Zhuo Xiaojuan Wang Zongxiu Sun 《Plant Cell, Tissue and Organ Culture》2010,102(2):163-170
Malaxis acuminata is a terrestrial orchid that grows in shady areas of semi-evergreen to shrubby forests. It is highly valued for its medicinal
properties as dried pseudo-bulbs are important ingredients of several Ayurvedic preparations. In this study, adventitious
shoot buds were induced from internodal explants of M. acuminata grown on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn),
and thidiazuron (TDZ). Of the three cytokinins used, TDZ at 3 mg l−1 induced the highest frequency (82%) of organogenic explants. However, all responding explants produced only a single adventitious
shoot irrespective of the type and concentration of the cytokinin. Adding 0.5 mg l−1 α naphthaleneacetic acid (NAA) to the medium enhanced adventitious shoot formation. In the presence of 3 mg l−1 TDZ and 0.5 mg l−1 NAA, frequency of organogenesis was 96% with a mean number of 6.1 shoots per explant. Prolonged culture or subculture on
the same medium did not promote further shoot production. However, transfer of these cultures to MS medium supplemented with
3 mg l−1 TDZ and 0.5 mg l−1 NAA and various concentrations of different polyamines (PAs), including spermine, spermidine, and putrescine, significantly
increased mean shoot number per explant. The highest frequency of shoot induction (100%) and mean shoot number per explant
(14.6) was observed on MS medium with 3 mg l−1 TDZ, 0.5 mg l−1 NAA, and 0.4 mM spermidine. Regenerated shoots were excised and subcultured on an elongation medium consisting of MS medium
with 3 mg l−1 BA. Moreover, the highest frequency of rooting (96%) and mean number of roots per shoot (3.3) was observed on MS medium with
4 mg l−1 indole-3-butyric acid (IBA) and 1.5 mg l−1 activated charcoal (AC). Almost 90% of rooted shoots were successfully acclimatized and established ex vitro. 相似文献
20.
Li-Hua Zhu Xiao-Qin Wu Hong-Ye Qu Jing Ji Jian-ren Ye 《Plant Cell, Tissue and Organ Culture》2010,102(1):121-128
A protocol was developed for the micropropagation of Pinus massoniana and mycorrhiza formation on rooted microshoots. Seedling explants were first cultured on Gresshoff and Doy (GD) medium supplemented
with 6-benzyladenine (BA) alone or in combination with α-napthaleneacetic acid (NAA) to stimulate the formation of intercotyledonary
axillary buds. The frequency of axillary bud induction was up to 97% on medium supplemented with 4.0 mg l−1 BA and 0. 2 mg l−1 NAA, and the average number of buds per explant reached up to 5.5 on medium with 4.0 mg l−1 BA and 0.1 mg l−1 NAA. Axillary buds elongated rapidly after being transferred to half-strength GD medium containing activated charcoal (0.1%
w/v). Shoot proliferation was achieved by cutting elongated shoots into stem segments and subculturing on GD medium containing
2 mg l−1 BA and 0.2 mg l−1 NAA. Root primordia were induced in 82% of shoots when transferred to half-strength GD medium containing 0.2 mg l−1 NAA. Root elongation was achieved in a hormone-free GD agar medium or a perlite substrate. Rooted plantlets were inoculated
with the mycelium of ectomycorrhizal fungus Pisolithus tinctorius and the formation of ectomycorrhiza-like structures was achieved in vitro. 相似文献