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1.
Chun-Miao Feng Rongda Qu Li-Li Zhou De-Yu Xie Qiu-Yun Xiang 《Plant Cell, Tissue and Organ Culture》2009,97(1):27-37
Dwarf dogwoods (or the bunchberries) are the only suffrutex in Cornaceae. They are attractive ground cover ornamentals with clusters of small flowers surrounded by petaloid bracts. Little has been
reported on plant regeneration of dogwoods. As a step toward unraveling the molecular basis of inflorescence evolution in
Cornus, we report an efficient regeneration system for a dwarf dogwood species C. canadensis through organogenesis from rejuvenated leaves, and characterize the development of the plantlets. We used the nodal stem
segments of vegetative branches as explants. Micropropogated shoots were quickly induced from axillary buds of nodes on an
induction medium consisting of basal MS medium supplemented with 4.44 μM BAP and 0.54 μM NAA. The new leaves of adventitious
shoots were used as explants to induce calli on the same induction medium. Nearly 65% of leaf explants produced calli, 80%
of which formed adventitious buds. Gibberellic acid (1.45 μM) added to the same induction medium efficiently promoted quick
elongation of most adventitious buds, and 0.49 μM IBA added to the basal MS medium promoted root formation from nearly 50%
of the elongated shoots. The growth of plantlets in pot soil was characterized by the development of functional woody rhizomes,
which continuously developed new aboveground vegetative branches, but not flowering branches, within the past 12 months. Potential
reasons causing the delay of flowering of the regenerated plants are discussed. The establishment of this regeneration system
facilitates developing a genetic transformation system to test candidate genes involved in the developmental divergence of
inflorescences in Cornus.
Electronic supplementary material The online version of this article (doi:) contains supplementary material, which is available to authorized users. 相似文献
2.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Pistacia vera L. cv. Siirt. The in vitro procedure involved four steps that included (1) induction of shoot initials from the regenerated mature leaf tissue, (2)
regeneration and elongation of shoots from the shoot initials, (3) rooting of the shoots, and (4) acclimatization of the plantlets.
The induction of shoot initials was achieved on an agarified Murashige and Skoog (MS) medium with Gamborg vitamins supplemented
in different concentrations of benzylaminopurine (BA) and indole-3-acetic acid (IAA). The best medium for shoot induction
was a MS medium with 1 mgl−1 IAA and 2 mgl−1 BA. Numerous shoot primordia developed within 2–3 wk on the leaf margin and the midrib region, without any callus phase.
In the second step, the shoot clumps were separated from the leaf explants and transferred to a MS medium supplemented with
1 mgl−1 BA, resulting in a differentiation of the shoot initials into well-developed shoots. The elongated shoots (>3 cm long) were
rooted on a full-strength MS basal medium supplemented with 2 mgl−1 of indole-3-butyric acid in the third stage. Finally, the rooted plants were transferred to soil with an 80% success rate.
This protocol was utilized for the in vitro clonal propagation of this important recalcitrant plant species. 相似文献
3.
Morphogenesis induction and organogenic nodule differentiation in <Emphasis Type="Italic">Populus euphratica</Emphasis> Oliv. leaf explants 总被引:1,自引:1,他引:0
Sílvia Ferreira Dora Batista Susana Serrazina Maria Salomé Pais 《Plant Cell, Tissue and Organ Culture》2009,96(1):35-43
Populus euphratica Oliv. is a deciduous poplar species, occurring mainly in riparian areas of China and Middle Eastern countries, and presenting
high tolerance to extreme temperatures and high soil salinity. In this study, an optimized protocol for development and propagation
of P. euphratica from leaf explants is reported, based on a morphogenic process that involves organogenic nodule differentiation. Adventitious
shoot regeneration of P. euphratica from organogenic nodules of leaf explants was achieved within a range of concentrations of α-naphtalenacetic acid and 6-benzylaminopurine,
at a fixed 2:1 ratio. Cambial cells started to divide 5 days after inoculation on culture medium and, after 12 days, several
organizing centres were already formed. Non-friable callus tissue, together with organization centres, formed structures that
evolved to nodules after about 40 days which were, then, able to regenerate new shoots after 50–60 days. The nodules did not
separate from the mother explants and were able to successfully give rise to new adventitious shoots. These were rescued and
successfully grown and rooted in different culture media, and fully developed plants were obtained. The regeneration system
here described for P. euphratica is innovative, reproducible and data from histological studies of the morphogenic process support the classification of the
regenerative structures as organogenic nodules. 相似文献
4.
B. Wang D. X. Peng Z. X. Sun N. Zhang S. M. Gao 《In vitro cellular & developmental biology. Plant》2008,44(2):105-111
Ramie [Boehmeria nivea (L.) Gaud] is one of the most important perennial fiber crops in China. In vitro tissue culture of ramie could serve as an important means for its improvement through genetic transformation. To improve
the regeneration capacity of ramie, the effects on plant regeneration of donor plant age, basal medium, plant growth regulators,
and culture conditions were evaluated using explants derived from the cotyledon, hypocotyl, leaf, petiole, and stem of ramie
seedlings. Cotyledons and hypocotyls excised from 4-d-old seedlings and leaves and petioles and stems from 15-d-old seedlings
were optimal explants. The highest regeneration efficiency was obtained on Murashige and Skoog salts with Gamborg’s B5 vitamins basal medium containing 2.27 μM thidiazuron (TDZ) and 0.054 μM naphthaleneacetic acid (NAA) for the five explant
types tested. A photoperiod of 16:8 h (light/dark) was found to be superior than continuous darkness for regeneration of ramie
using TDZ. The regenerated shoots were transferred to hormone-free medium for shoot elongation and successfully rooted on
half-strength Murashige and Skoog supplemented with 0.134 μM NAA. The rooted plantlets with four to five leaves were transplanted
to greenhouse for further growth. 相似文献
5.
A successful micropropagation system was developed for four different medicinal Maesa species. Multiple shoots were induced through both axillary bud formation and adventitious shoot regeneration from leaf explants.
The explants were cultured on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA), thidiazuron (TDZ) and/or
α-naphthalene acetic acid (NAA). The success of regeneration varied for different species and depended on the type and concentration
of plant growth regulators. Regenerated shoots spontaneously developed roots within 6 weeks on MS hormone-free medium. The
rooted shoots were transferred to the greenhouse with a 100% success rate. Furthermore, flow cytometry analysis indicated
that there were no changes in ploidy level of those regenerated shoots as compared with wild type adult plants. Thin layer
chromatography (TLC) analysis revealed that common and distinguishing spot of saponins were similarly observed in regenerated
shoots compared to the control plants. Therefore, the protocol also provides an effective means for the in vitro conservation
of Maesa spp. that produce pharmaceutically interesting saponins. 相似文献
6.
In vitro regeneration protocol for Anethum graveolens (Apiaceae) was developed using leaf explants. MS basal medium used in experiments was augmented with various hormones for
caulogenic and rhizogenic response. The optimum callus induction (100%) was obtained by leaf explants on MS media fortified
with BA (0.5 mg l−1) singly and in combination with NAA (0.1 and 0.2 mg l−1). BA at 0.5 mg l−1, KN at 1.0 mg l−1 and NAA at 0.1 mg l−1 induced highest number of multiple shoots (10.0 ± 0.25) per explant and they also showed in vitro flowering within 3 weeks
of culture. Influence of adenine sulfate on regeneration frequency of callus was evaluated. The highest frequency of rooting
(100%) with 6.0 ± 0.25 roots per explants was obtained in one-fourth strength MS medium supplemented with 1/4 MS + IBA 0.5 mg l−1 within 4 weeks of transfer to the rooting medium. In vitro flowering (35%) was obtained with MS fortified with BA alone and
also in combination with KN and NAA (5.3 ± 0.42 flowers per explants). In vitro flowering response was tested with different
carbohydrates (fructose, glucose, mannose and sorbitol) and optimized. Hardening was successfully attained under controlled
conditions inside the plant tissue culture room. The proposed method could effectively be applied for the conservation and
clonal propagation to meet the pharmaceutical demands of this medicinally important species. 相似文献
7.
Leafy spurge (Euphorbia esula L.) is a perennial, invasive weed used as a model to study invasive plant behavior, because molecular tools (such as a deep
expressed sequence tag database and deoxyribonucleic acid microarrays) have been developed for this species. However, the
lack of effective in vitro regeneration and genetic transformation systems has hampered molecular approaches to study leafy spurge. In this study, we
describe an efficient in vitro regeneration system. Three highly regenerative lines were selected by screening the in vitro regeneration capabilities of stem explants of 162 seedlings. The effects of various culture conditions on in vitro regeneration were then evaluated based on explant competence to form calluses and shoots. High rates of shoot regeneration
can be obtained using a growth medium containing 1× woody plant basal medium and 1× Murashige and Skoog (MS) basal salts,
1× MS vitamins, 1.11 μM 6-benzylaminopurine, 1.97 μM indole-3-butyric acid, and 3% sucrose, pH 5.6–5.8. After 30 d culture,
multiple shoots formed either directly from the stem or indirectly from the callus. This method is a requisite for the development
of genetic transformation systems for leafy spurge and may be used to develop in vitro regeneration techniques for other species in the Euphorbiaceae. 相似文献
8.
Adult plants are known for recalcitrance when it comes to adventitious organ formation and regeneration. Methods used for
regeneration in explants from seedlings of Campanula carpatica failed to work for explants from adult plants of the same species. The present investigation generated efficient regeneration
methods for mature specimens of four species of Campanula, C. carpatica, C. haylodgensis, C. portenschlagiana and C. poscharskyana. Petiole explants from dark-grown in vitro shoot cultures grown from nodal cuttings of adult plants regenerated successfully
(95%), while explants from light-grown in vitro shoot cultures and greenhouse-grown plants regenerated at 12% and zero percentage,
respectively. Dark-treatment, along with media manipulation with plant growth regulators, further enhanced regenerative capacity
of the explants. A MS-based medium containing 10mg l −1 TDZ and 0.25 mg l−1 NAA was the most efficient regeneration medium. Transgenic shoots from C. carpatica (3%) and C. haylodgensis (1%) and transgenic callus from all species were produced using Agrobacterium tumefaciens, and transformation was confirmed by histochemical and Southern blot analyses. Protocols developed in this study may be useful
for achieving efficient regeneration and transformation of recalcitrant adult plants. 相似文献
9.
In vitro micropropagation by direct organogenesis and somatic embryogenesis via callus was developed for Crambe tataria (Brassicaceae). C. tataria is an endemic species of the Pontic-Pannonic region, but it is also present in Italy, where it is localized in Friuli on
a characteristic grassland formation, called “magredi”. C. tataria is regarded as an endangered species. Leaf and root explants were subjected to plant regulator treatments, which invoked
different morphogenic responses. Leaf explants produced more callus than root explants and a higher amount of callus was obtained
with 1 mg l−1 2,4-D in combination with 2 mg l−1 Kin. Somatic embryogenesis was obtained in calli maintained in a delayed subculture regime on media containing BAP in combination
with NAA. Root explants cultured with BAP combined with NAA developed adventitious rosette shoots. Shoots rooted on half-strength
MS media, and the number of roots per plantlet and their length were heavily dependent on sucrose content. The in vitro regenerated
plantlets were acclimatized ex vitro and a mean of 50% of the plantlets survived and showed a true-to-type growth habit. This
study describes the development of two in vitro micropropagation protocols, via direct organogenesis and via embryogenesis
from callus, that are the basis for the application of in vitro tools for the establishment of basal collections with representative
genetic diversity and for the long-term storage of plant genetic material. 相似文献
10.
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
11.
An efficient transformation protocol was developed for Eucalyptus tereticornis Sm. using cotyledon and hypocotyl explants. Precultured cotyledon and hypocotyl explants were cocultured with Agrobacterium tumefaciens strain LBA 4404 harboring the binary vector pBI121 containing the uidA and neomycin phosphotransferase II genes for 2 d and transferred to selective regeneration medium containing 0.5 mg/l 6-benzylaminopurine
(BAP), 0.1 mg/l naphthalene acetic acid, 40 mg/l kanamycin, and 300 mg/l cefotaxime. After two passages in the selective regeneration
medium, the putatively transformed regenerants were transferred to Murashige and Skoog (MS) liquid medium containing 0.5 mg/l
BAP and 40 mg/l kanamycin on paper bridges for further development and elongation. The elongated kanamycin-resistant shoots
were subsequently rooted on the MS medium supplemented with 1.0 mg/l indole-3-butyric acid and 40 mg/l kanamycin. A strong
β-glucuronidase activity was detected in the transformed plants by histochemical assay. Integration of T-DNA into the nuclear
genome of transgenic plants was confirmed by polymerase chain reaction and southern hybridization. This protocol allows effective
transformation and direct regeneration of E. tereticornis Sm. 相似文献
12.
Houcheng Zhou Ming Li Xia Zhao Xiucai Fan Aiguang Guo 《Plant Cell, Tissue and Organ Culture》2010,101(1):79-87
A complete protocol for adventitious shoot regeneration was developed from the leaves of peach rootstock ‘Nemaguard’(Prunus persica × P. davidiana) grown in vitro. Shoot explants were cultured in vitro in Murashige and Skoog medium supplemented with 3.55 μM 6-benzyladenine
and 7.38 μM indole-3-butyric acid (IBA). Non-expanded leaves along with their petioles from 3-week-old in vitro-grown shoots
were used as explants. Regeneration percentage was influenced by plant growth regulators, basal medium, explant type, dark
period, and gelling agents. Optimal regeneration was observed with leaf explants wounded by transverse cuts twice along the
midrib and first incubated with abaxial surfaces facing upward in the dark for 3 weeks, and then transferred to the light
and cultured with the adaxial side in contact with regeneration medium, as seen on 1/2 MS, woody plant medium or Schenk and
Hildebrandt medium supplemented with 9.08 μM thidiazuron, 0.54 μM IBA and 0.25% agar. This produced the highest regeneration
percentage at 71.7% and a mean of 5.74 ± 3.24 shoots on 1/2 MS medium. Adventitious shoots were rooted (98.3–100%) and rooted
plantlets survived after acclimatization to the greenhouse. 相似文献
13.
An efficient system for shoot regeneration and Agrobacterium-mediated gene transfer into Brassica napus was developed through the modification of the culture conditions. Different concentrations of benzyladenine (1.5, 3.0 and 4.5 mg dm–3) and thidiazuron (0.0, 0.15 and 0.30 mg dm–3) were evaluated for shoot regeneration of 7, 14 and 21-d-old hypocotyl explants. Maximum shoot regeneration frequency was obtained in 21-d-old explants using 4.5 mg dm–3 benzyladenine and 0.3 mg dm–3 thidiazuron. Under above culture condition, the highest percentage of shoot regeneration frequency was 200 %. Agrobacterium-infected explants grown on the selection medium gave rise to transgenic shoots at a frequency of 11.8 %. Transformed shoots rooted when cultured on a medium supplemented with 2 mg dm–3 of indolebutyric acid and 10 mg dm–3 kanamycin. The rooted plantlets were successfully established in the soil and developed fertile flowers and viable seeds. Evidences for transformation were confirmed by GUS assay and PCR analysis. 相似文献
14.
Tantravahi Srinivasan Koppolu Raja Rajesh Kumar Pulugurtha Bharadwaja Kirti 《Plant Cell, Tissue and Organ Culture》2010,101(3):303-309
Though peanut tissue culture has advanced to a considerable extent using a number of explants with the subsequent production
of transgenic plants, wild Arachis species appeared to be recalcitrant to using similar explants. In this study, the use of cotyledonary nodes as explants prepared
from 7-day old seedlings resulted in the development of a simple and rapid regeneration protocol for five diploid wild species
including A. diogoi, A. stenosperma, A. duranensis, A. cardenasii and A. correntina belonging to the genus Arachis for producing multiple shoots. Shoot bud initiation was observed 10 to 15 days after culture initiation. Responding cotyledonary
nodes with shoot buds were subcultured to lower levels of cytokinin and finally to MS basal medium for further shoot development
and elongation. Production of multiple shoots was observed in all the five diploid species with a maximum of 9 to 16 shoots
were obtained per explant in the primary cultures. The number of shoot buds increased significantly with repeated explant
subculturing with recovery up to 45 shoots from responding explants. These shoots were rooted efficiently on MS medium supplemented
with 1 mg l−1 naphthalene acetic acid and the time taken from explanting to the transfer of shoots to potting mixture was about 12 weeks.
All rooted shoots were successfully established in soil in glass house and further transferred to field. These plants survived
to maturity and set seed. 相似文献
15.
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
16.
In this study, we established an in vitro regeneration system to maximize the recovery of leafy perilla (Perilla frutescens L. Britton) plantlets as part of developing a molecular biotechnology-based metabolic engineering program for this crop plant.
Hypocotyl segments including the apical buds were used as explants for the direct production of shoots without an interim
callus phase. The number of shoots produced from the apical buds peaked within 3–4 weeks, and the shoots were subsequently
cultured on Murashige and Skoog (MS) media supplemented with 2 mg l−1 benzylaminopurine (BA). Spontaneous rhizogenesis was observed after 7–10 days of culture on MS media without hormonal additives.
The rooted shoots developed into normal plants in soil after hardening on distilled water for 3–4 days. The average plantlet
regeneration frequency was higher for the apical buds (64.33%) than for the top (15.66%), middle (4%), and basal (1.33%) segments
of the hypocotyls. This regeneration system demonstrates a capacity for high-frequency plantlet recovery and thus should be
considered for use in the genetic manipulation of leafy perilla. 相似文献
17.
Angela Carra Maurizio Sajeva Loredana Abbate Mirko Siragusa Francesco Sottile Francesco Carimi 《Plant Cell, Tissue and Organ Culture》2012,109(2):373-381
A new technique to regenerate caper plants (Capparis spinosa L. subsp. rupestris) starting from flower explant is reported. In vitro plant regeneration was attempted using stigma, anthers and unfertilized
ovules of unopened flowers collected in the field. Plant regeneration was achieved from unfertilized ovules on MS medium supplemented
with 88 mM sucrose and 13 μM 6-benzyladenine (BA). New individuals obtained from unfertilized ovules were used as source material
for micropropagation and multiple shoots were obtained on MS medium supplemented with the adeninic cytokinin BA and the auxin
indole-3-butyric acid (IBA). Explants obtained in micropropagation step were used for rooting step under several treatments.
The best results (100% of rooted explants) were obtained when explants were dipped for 10 min in 50 μM IBA solution and successively
maintained in growth regulator free medium. New plants were vigorous, of good quality and presented phenotypic characters
similar to mother plants. Furthermore genetic stability of regenerants was verified through flow cytometric analysis and two
different DNA-based techniques. 相似文献
18.
In vitro propagation protocol for Lilium oxypetalum, a high altitude Himalayan endemic lily, has been developed. Effect of explant types (i.e., callus and in vitro bulblet scales)
and sucrose concentration [0–6.0% (w/v)] on in vitro bulblet regeneration of L. oxypetalum was tested in previously optimized Murashige and Skoog basal medium supplemented with 2.0 μM 6-benzyladenine and 0.1 μM α-naphthaleneacetic acid. Callus explants produced significantly (P < 0.01) higher number of bulblets per explant than bulblet scale explants. Of the different concentrations of sucrose tested,
4.5% (w/v) sucrose showed significantly (P < 0.01) higher percentage regeneration (i.e., 70.8 ± 4.2 and 79.2 ± 4.2% regeneration on callus and bulblet scale explants,
respectively), and produced higher number of bulblets per explant (i.e., 9.0 ± 0.4 and 5.4 ± 0.5 bulblets on callus and bulblet
scale explants, respectively). Regenerated bulblets developed 2–3 leaves when subcultured for 4 weeks and were subsequently
transferred ex vitro with a survival rate of 66.7% after 6 weeks. Leaves of the survived plantlets became dry after growing
ex vitro for 10 weeks, amongst which 86.4% re-sprouted after remaining dormant for 5–6 weeks and produced 1.5 bulblets per
explant. Findings of the present study hold promise for efficiently multiplying the target species in view of its potential
economic and conservation significance. 相似文献
19.
Boon Chin Tan Chiew Foan Chin Peter Alderson 《Plant Cell, Tissue and Organ Culture》2011,105(3):457-463
An indirect in vitro plant regeneration protocol for Vanilla planifolia has been established. Juvenile leaf and nodal segments from V. planifolia were used as explants to initiate callus. Nodal explants showed better callus initiation than juvenile leaf explants, with
35.0% of explants forming callus when cultured on Murashige and Skoog (MS) basal medium supplemented with 2.0 mg/l 1-naphthylacetic
acid (NAA) and 1.0 mg/l 6-benzyladenine (BA). Almost 10.0% of juvenile leaf explants were induced to form callus on the MS
basal medium containing 2.0 mg/l NAA and 2.0 mg/l BA, whereas no callus formed in the presence of any concentrations of 2,4-dichlorophenoxyacetic
acid (2,4-D) and BA. After 8 weeks, callus generated was transferred to MS basal medium containing 1.0 mg/l BA and 0.5 mg/l
NAA. A mean number of 4.2 shoots per callus was produced on this medium, with a mean length of 3.8 cm after 8 weeks of culture.
Roots formed on 88.3% of plantlets when they were cultured on MS medium supplemented with 1.0 mg/l NAA, with a mean length
of 4.4 cm after 4 weeks of culture. Of the rooted plantlets, 90.0% survived acclimatisation and were making new growth after
4 weeks. 相似文献
20.
Shyamkumar Barampuram Byung Yeoup Chung Seung Sik Lee Byung Chull An Eun Mi Lee Jae-Young Cho 《In vitro cellular & developmental biology. Plant》2009,45(2):155-161
The present study demonstrates the establishment of embryogenic tissue from seeds and (seedling-derived hypocotyls) shoot
base explants derived from seedlings of Eremochloa ophiuroides. The highest percentage of callus induction obtained from seed and young shoot base explants was 52.0% and 66.6% on Murashige
and Skoog (MS) basal media supplemented with 9.0 μM and 18.1 μM 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. The
type of callus obtained from both types of explants was off-white to yellow in color and non-friable and shiny in texture.
Excised callus from the explants was subcultured onto fresh media of the same recipe for further proliferation. After 10–12 d
of subculture, a yellow, globular, friable embryogenic callus was obtained from the initial callus. The highest percentage
of embryogenic calli obtained at 40.0% was observed on media containing 2.2 μM 2,4-D. The highest regeneration rate of 46.6%
was observed on MS media supplemented with 0.4 μM 2,4-D and 2.2 μM benzylaminopurine (BA). Regenerated shoots were rooted
in MS basal medium. Plants with well-developed roots were transferred to pots containing a soil mix and acclimatized in greenhouse
conditions. Four weeks post-transfer, acclimatized plants showed 100% survival and remained healthy and green. This is the
first report of a successful method for induction of somatic embryogenesis with subsequent plant regeneration in centipede
grass and demonstrates the establishment of embryogenic callus and efficient plant regeneration with potential application
in the development of genetic transformation systems for centipede grass. 相似文献