共查询到20条相似文献,搜索用时 478 毫秒
1.
Veena Agrawal Pratima Rani Sardar 《In vitro cellular & developmental biology. Plant》2007,43(6):585-592
In vitro regeneration through somatic embryogenesis as well as organogenesis using cotyledon of a woody medicinal legume, Cassia angustifolia is reported. The cotyledons dissected from semi-mature seeds, if inoculated on Murashige and Skoog’s medium (MS) supplemented
with auxin alone or in combination with cytokinin, produced direct and indirect somatic embryos. A maximum of 14.36 ± 2.26
somatic embryos per 20 mg of explants including callus were produced in 70% cultures on MS medium with 2.5 μM benzyladenine
(BA) + 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Although the percentage of embryogenic cultures was higher (83.33%) at
10 μM 2,4-D + 1 μM BA, the average number of somatic embryos was much less (7.6 ± 0.85) at this level, whereas at 2.5 μM BA
and 5 μM 2,4-D, there was a simultaneous formation of both somatic embryos and shoots. The somatic embryos, although started
germinating on the same medium, developed into full plantlets only if transferred to MS basal with 2% sucrose. Cytokinins
alone did not induce somatic embryogenesis, but formed multiple shoots. Five micromolar BA proved optimum for recurrently
inducing shoots in the competent callus with a maximum average of 12.04 ± 2.10 shoots and shoot length of 2.26 ± 0.03 cm.
Nearly 91.6% shoots (2–2.5 cm in size) organized an average of 5.12 ± 0.58 roots on half strength MS + 10 μM indole-3-butyric
acid. All the plantlets have been transferred successfully to soil. Types of auxin and its interaction with cytokinin significantly
influenced somatic embryogenesis. 相似文献
2.
Micropropagation of the endangered <Emphasis Type="Italic">Kniphofia leucocephala</Emphasis> Baijnath 总被引:3,自引:0,他引:3
Summary The species, Kniphofia leucocephala is extant at only one location, Langepan, KwaZulu-Natal in South Africa, where the population is threatened by afforestation
and possibly grazing. Consequently, a continuous culture system was established as part of a program for the propagation and
re-introduction of plants into the wild. The efficiency of the system in terms of shoot multiplication and, particularly,
the frequency and rate of root initiation was strongly influenced by the concentration of benzyladenine in the shoot multiplication
medium. The optimum shoot multiplication medium for subsequent root initiation contained 2 mgl−1 (8.9 μM) benzyladenine alone. The shoots were successfully rooted and acclimatized. Approximately 200 shoots can be produced from
one shoot after five 4-wk cycles. Thus, large numbers of plantlets can be propagated in this continuous culture system, serving
conservation interests. 相似文献
3.
Summary Efficient and highly reproducible induction of somatic embryogenesis was obtained in four out of seven selected clones of
neem, Azadirachta indica A. Juss. This was achieved either directly from root and nodal explants or indirectly from callus cultures initiated from
leaf explants excised from 1-yr-old axenic plants. Direct induction of somatic embryogenesis was achieved both from nodal
and root segments within 8 wk of culture on MS1 medium without growth regulators. However, the addition of 2.3–4.5 μM thidiazuron and 0.5 μM 2,4-dichlorophenoxyacetic acid into the medium were necessary to induce somatic embryogenesis via callus phase from leaf
explants. Repetitive embryogenesis was observed within 3–4 wk following transfer of somatic embryos to a plant growth regulator-free
medium. When somatic embryos of nodal and root segments were left on the induction medium without subculturing, approximately
15% of the somatic embryos developed into whole plantlets after passing through a series of developmental stages. Plantlets
thus produced were hardy, lush green, and acclimatized casily under greenhouse conditions. However, somatic embryos derived
from leaf explants showed low conversion rates (<5%). HPLC analysis revealed no detectable levels of azadirachtin in somatic
embryos. 相似文献
4.
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
5.
Summary An efficient procedure for the in vitro propagation and cryogenic conservation of Syzygium francissi was developed. The maximum number of shoots per explant was obtained on a Murashige and Skoog (MS) medium supplemented with
4.5 μM benzyladenine and 0.5 μM indole-3-butyric acid (IBA). The in vitro-propagated shoots produced roots when transferred to MS medium containing IBA, indold-3-acetic acid, or naphthaleneacetic
acid at various concentrations. Rooted microshoots were transferred to a coco-peat, perlite, and vermiculite (1∶1∶1) mixture,
and hardened off under greenhouse conditions. Ninety-five percent of rooted shoots successfully acclimatized in the greenhouse.
Shoot tips excised from in vitro-grown plants were successfully cryostoraged at −196°C by the encapsulation-dehydration method. A preculture of formed beads
on MS medium containing 0.75 M sucrose for 1 d, followed by 6 h dehydration (20% moisture content) led to the highest survival rate after cryostorage for
1h. This method is a promising technique for in vitro propagation and cryopreservation of shoot tips from in vitro-grown plantlets of S. francissi germplasm. 相似文献
6.
Summary Somatic embryo (bipolar) or shoot (monopolar) morphogenesis in mesophyll cells of Euphorbia nivulia Buch.-Ham in vitro was dependent on the type of auxin supplementing Murashige and Skoog (MS) medium containing benzyladenine. Direct in vitro morphogenesis, i.e., organogenesis, and somatic embryogenesis were significantly influenced by seasonal growth of the donor
plant, explant position (proximal, mid, and distal), and light. Explants collected in march/April were superior to July/August
material. Proximal explants underwent morphogenesis more readily than mid- and tip-derived explants. Incubation in the light
favored morphogenesis while darkness was inhibitory. Kinetin (Kn) was also inhibitory to morphogenesis. MS medium enriched
with different levels of N6-benzyladenine (BA) alone, or in combination with α-naphthaleneacetic acid (NAA) or indole-3-acetic acid (IAA), induced adventitious
shoots directly. Explants collected in March/April cultured on medium with 13.3 μM BA and 2.69 μM NAA developed the highest number of shoots, a mean of 15.2 shoots per proximal explant. Developed shoots rooted the best
on half-strength MS medium with 2.46 μM indole-3-butyric acid, which developed a mean of 5.2 roots per shoot. Rooted healthy shoots could be transplanted to small
pots, with an 80% survival rate. Addition of 2,4-dichlorophenoxyacetic acid (2.4-D) to BA-supplemented medium was obligatory
to develop somatic embryos. MS medium containing 2.26 μM 2,4-D and 4.44 μM BA induced a mean of 44.8 somatic embryos per proximal explant. The embryos passed through distinct stages of embryogenesis,
namely globular, heart, torpedo, and early cotyledonary. The embryos (88%) underwent maturation on half-strength MS medium
with 2.89 μM gibberellic acid (GA3), and its subsequent transfer on half-strength MS basal medium in light conditions facilitated 80% conversion of embryos
to plantlets. Direct shoots or embryos were originated from the mesophyll cells. Somatic embryo development was concurrent
with the independent origin of vasculature in the bulbous basal portion. The survival rate of embryo-derived plants was 90%. 相似文献
7.
Elif Aylin Ozudogru Ergun Kaya Emrah Kirdok Saliha Issever-Ozturk 《In vitro cellular & developmental biology. Plant》2011,47(2):309-320
An efficient in vitro propagation protocol, applicable both to young and mature explants of two Thymus spp., results in genetically stable plantlets. In vitro-grown shoot tips of Thymus vulgaris L. were exposed to cytokinins (6-benzyladenine, kinetin, and thidiazuron) alone or in combination with auxins, gibberellic
acid (GA3) and/or silver nitrate in order to optimize in vitro shoot proliferation. Optimum shoot proliferation (97% regeneration rate, with 8.6 shoots produced per explant) was obtained
when semi-solid Murashige and Skoog (MS) medium was supplemented with 1 mg L−1 kinetin and 0.3 mg L−1 GA3. Rooting of the shoots was easily obtained on semi-solid MS medium that was either hormone-free or supplemented with auxins.
However, the best root apparatus (92.5% rooting rate, with 19 adventitious roots per shoot) developed on MS medium supplemented
with 0.05 mg L−1 2,4-dichlorophenoxyacetic acid. Genetic stability was confirmed in the in vitro-germinated mother plant as well as the shoots that underwent two, four, six, eight, or ten cycles of in vitro subculturing by random amplified polymorphic DNA (RAPD) analysis. When applied to the micropropagation of mature shoot tips
of T. longicaulis C. Presl subsp. longicaulis var. subisophyllus (Borbás) Jalas, the optimized in vitro propagation protocol resulted in a 97.5% shoot regeneration rate, with five shoots formed per explant, and 100% rooting.
Rooted plantlets of both species were transferred to 250-mL plastic pots and successfully acclimatized by gradually reducing
the relative humidity. 相似文献
8.
Summary Efficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth
regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and
Skoog (MS) medium supplemented with 4.52μM2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior
and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52μM 2,4-D to suspension cultures of half- or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 μM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous
with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength
MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets
acclimatized under field conditions with 90% survival. 相似文献
9.
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
10.
Branka Vinterhalter Dijana Krstić Milošević Teodora Janković Jelena Milojević Dragan Vinterhalter 《Central European Journal of Biology》2012,7(4):690-697
Gentiana dinarica Beck, rare and endangered species of Balkan Dinaric alps, was in vitro propagated (micropropagated) from axillary buds of plants collected at Mt. Tara, Serbia. G. dinarica preferred MS to WPM medium, with optimal shoot multiplication on MS medium with 3% sucrose, 1.0 mg l−1 BA and 0.1 mg l−1 NAA. Rooting was not clearly separated from shoot multiplication since BA did not completely inhibit root initiation. Spontaneous
rooting on plant growth regulator-free medium occurred in some 30% of shoot explants. Rooting was stimulated mostly by decreased
mineral salt nutrition and a medium with 0.5 MS salts, 2% sucrose and 0.5–1.0 mg l−1 IBA was considered to be optimal for rooting. Rooted plantlets were successfully acclimated and further cultured in peat-based
substrate. 相似文献
11.
Veronique Troch Stefaan Werbrouck Danny Geelen Marie-Christine Van Labeke 《Plant Cell, Tissue and Organ Culture》2009,98(1):115-123
Factors affecting conversion of horse chestnut (A. hippocastanum L.) somatic embryos into plantlets were evaluated. Anther filament derived embryogenic tissue developed bipolar structures
with two cotyledons and a well-developed shoot and root apical meristem upon auxin omittance from the culturing medium. The
impact of carbohydrate type (glucose, fructose, sucrose and maltose) and concentration (3 and 6%) on somatic embryo maturation
and conversion were evaluated. Although conversion frequencies were high for all treatments, overall quality of regenerated
plantlets was poor. Increasing the carbohydrate concentration in the maturation medium did not increase conversion of somatic
embryos or quality of regenerated plantlets in terms of shoot height. On the contrary, addition of PEG (polyethylene glycol)
in maturation media had a beneficial effect on shoot quality of regenerated plantlets. Sucrose was a superior carbon source
when PEG was included in the maturation medium, in terms of conversion rate (65.7%) as well as of shoot quality of plantlets
(43.8% of plantlets had shoots >2 cm). Clonal fidelity of the different development stages of somatic embryogenesis and of
converted plantlets was assessed by flow cytometry and no major ploidy changes were found. 相似文献
12.
Laura Y. Solís-Ramos Sara Nahuath-Dzib Antonio Andrade-Torres Felipe Barredo-Pool Tomas González-Estrada Enrique Castaño de la Serna 《Biologia》2010,65(3):504-511
Capsicum chinense is recalcitrant in in vitro morphogenesis. No efficient, reproducible somatic embryogenesis regeneration system exists for this species, impeding regeneration
from transformed cells. An indirect somatic embryogenesis protocol is developed using mature C. chinense zygotic embryo segments (ZES). The ZES cultured in semi-solid Murashige-Skoog (MS) medium supplemented with 8.9 μM naphthaleneacetic
acid, 11.4 μM indoleacetic acid and 8.9 μM 6-benzylaminopurine, developed an embryogenic callus and 8% of the calli developed
somatic embryos. Torpedo-stage somatic embryos were detached from the callus and subcultured in semi-solid MS medium without
growth regulators, producing a 75% conversion rate to plantlets with well-formed root tissue. Histological analysis showed
the developed structures to have no vascular connection with the callus and to be bipolar, confirming that this protocol induced
formation of viable somatic embryos from mature C. chinense ZES. All acclimated plantlets survived under greenhouse conditions. This protocol will facilitate regeneration of genetically
transformed plants using either biolistics or Agrobacterium tumefaciens approach. 相似文献
13.
In vitro culture is an important aid for ex situ conservation of rare, endemic or threatened plants. In this work, we establish an efficient method for the seed germination,
seedling development, and axillary shoot propagation of Centaurea zeybekii Wagenitz. The seeds, collected from a wild population, were surface sterilised and cultured on various in vitro germination media. The effects of photoperiod and temperature on seed germination were also investigated. Germinations were
obtained after 6 weeks in culture and the radicle emergence was evaluated as a main indicator. A high frequency of germination
was obtained on distilled water supplemented with vitamines and 1 mg/L GA3. Although the seed germination frequencies were not affected by photoperiod, the highest germination frequency was obtained
at 24 ± 2°C. A high frequency of axillary shoot proliferation was produced on MS medium supplemented with 1 mg/L BA. Then,
the axillary shoots were separated and transferred to MS medium with or without plant growth regulators for rooting. Rhizogenezis
was promoted after 6 weeks only in MS and 1/2 MS media containing 0.5 mg/L IBA. The rooting process was very slow and the
percentage of shoot rooting was also very low (15%).
The present study not only enables reinforcement of wild plant populations using ex situ growth of individuals, but it also helps to large number of aseptic seedling to use it in clonaly micropropagation studies. 相似文献
14.
P. Giridhar E. P. Indu K. Vinod A. Chandrashekar G. A. Ravishankar 《Acta Physiologiae Plantarum》2004,26(3):299-305
For the first time direct somatic embryogenesis from hypocotyl explants of in vitro regenerated plantlets of C. arabica and C. canephora was achieved on modified MS medium containing 10 – 70 μM silver nitrate supplemented with 1.1 μM N6 benzyladenine and 2.85 μM indole-3-acetic acid. A maximum of 144.1±7.3 and 68.7±3.3 embryos per explant were produced at
40 μM silver nitrate in C. canephora and C. arabica respectively. Only yellow friable embryogenic callus obtained from the cut edges of most of leaf explants of both C. arabica and C. canephora at all concentrations of silver nitrate were tried in this experiment. Formation of secondary embryos from stage I primary
embryos (small yellow, round, globular embryos) was more (28.23±1.3) at 60 μM silver nitrate in C. canephora, while 40 μM silver nitrate supported more of secondary embryo formation in C. arabica (40.5±1.2). When stage II (green globular round matured embryos) and stage III primary embryos (tubular stage embryos) were
used, secondary embryo formation was very small and many of these embryos developed into plantlets and some of them even rooted.
By using these protocols within 45 – 60 days it is possible to get secondary embryos from primary embryos and direct somatic
embryos from hypocotyls of in vitro plantlets in both these Coffea species. 相似文献
15.
Filipendula ulmaria (L.) Maxim (meadowsweet) is a medicinal plant that is claimed to have several biological activities, including anti-tumor,
anti-carcinogenic, anti-oxidant, anti-coagulant, anti-ulcerogenic, anti-microbial, anti-arthritic, and immunomodulatory properties.
This report describes, for the first time, an efficient plant regeneration system for F. ulmaria via adventitious shoot development from leaf, petiole, and root explants cultured on Murashige and Skoog’s minimal organics
medium containing different concentrations of thidiazuron (TDZ), benzyladenine, and kinetin either alone or in combination
with different auxins. Relatively extensive/prolific shoot regeneration was observed in all three explant types with TDZ in
combination with indole-3-acetic acid (IAA). Gibberellic acid (GA3), TDZ, and IAA combinations were also tested. The best shoot proliferation was observed among root explants cultured on media
supplemented with 0.45 μM TDZ + 2.85 μM IAA + 1.44 μM GA3. Regenerated shoots were transferred to rooting media containing different concentrations of either IAA, indole-3-butyric
acid (IBA), naphthalene acetic acid, or 2,4-dichlorophenoxyacetic acid. Most shoots developed roots on medium with 2.46 μM
IBA. Rooted explants were transferred to vermiculite in Magenta containers for a 2-wk acclimatization period and then finally
to plastic pots containing potting soil. The plantlets in soil were kept in growth chambers for 2 wk before transferring to
greenhouse conditions. 相似文献
16.
Semiha Erisen Mustafa Yorgancilar Emine Atalay Mehmet Babaoglu 《Plant Cell, Tissue and Organ Culture》2010,100(2):229-233
Prolific shoot regeneration via organogenesis was induced from leaf and leaf petiole explants of the endemic Astragalus cariensis species on Murashige and Skoog (MS) medium with α-naphthaleneacetic acid (NAA) and benzyladenine (BA) within 8 week. The
highest number of shoots (23/explants) was obtained from leaf explants cultured on MS with 0.5 mg/l NAA and 4 mg/l BA. Elongated
shoots were successfully rooted in MS medium with 0.5 mg/l indole-3-butyric acid. Rooted plantlets were acclimatized in pots
containing 1:1 mixture of peat and perlite. 相似文献
17.
Summary Plantlets of Capsicum annuum L. ev. Sweet Banana regenerated via somatic embryogenesis from immature zygotic embryos were capable of producing flower,
fruit, and seed when cultured in small tissue culture containers. In vitro floral buds were first formed on plantlets that grew on plantlet development medium [agar-gelled Murashige and Skoog (MS)
basal medium containing 1 mgl−1 (5.3 μM) α-naphthaleneacetic acid (NAA)] in a growth room at 22°C and continuous illumination. However, floral buds rarely developed
further into mature flowers. This problem was overcome using the vented autoclavable plant tissue culture containers. In vitro fruit formation and ripening was observed when liquid half-strength MS basal medium supplemented with 5 μg ml−1 silver thiosulfate, 1 mg l−1 (5.3 μM) NAA, and 3% sucrose was added to the surface of the plantlet development medium. Hand-pollination improved fruit set. Further
research in needed to determine why the pepper seeds formed in vitro failed to germinate. 相似文献
18.
Summary
In vitro propagation of Pelecyphora aselliformis, a Mexican cactus which is considered rare and is highly valued in the commercial market, was initiated using seeds as explants.
The longitudinal explants from seedlings germinated in vitro were cultivated on Murashige and Skoog medium containing 8.8 μM benzyladenine (BA) or 4.6 μM kinetin at pH 7.0. After 120 d, each explant gave rise to five shoots and this number of shoots increased 20–25% after subculture.
The hyperhydricity was similar in both media, but callus formation was lower on the medium with BA. The shoot development,
in terms of epicotyl length, and fresh and dry weight after 6 wk, was also recorded. The epicotyl length was similar on shoot-forming
media but the quality of shoots was better on media containing BA. In about 1 yr, 500–600 well-defined shoots were obtained.
The rooting of shoots was very slow and a vigorous radical system was observed after 1 yr of culture. 相似文献
19.
Summary
Kalopanax pictus (Thunb.) Nakai is a tall tree, and its wood has been used in making furniture, while its stem bark is used for medicinal
purposes. Here, we report on the micropropagation of Kalopanax pictus via somatic embryogenesis. Embryogenic callus was induced from immature zygotic embryos. The frequency embryogenic callus
induction is influenced by days of seed harvest. Callus formation was primarily observed along the radicle tips of zygotic
embryos incubated on Murashige and Skoog (MS) medium with 4.4 μM 2,4-dichlorophenoxyacctic acid (2,4-D). Somatic embryogenesis was observed following transfer of embryogenic callus to MS
medium lacking 2,4-D. Somatic embryos at the cotyledonary stage were obtained after 6 wk following culture. Frequency of conversion
of somatic embryos into plantlets was low (35%) on a hormone-free MS basal medium, but it increased to 61% when the medium
was supplemented with 0.05% charcoal. Gibberellic acid (GA3) treatment markedly enhanced the germination frequency of embryos up to 83%. All plantlets obtained showed 98% survival on
moist peat soil (TKS2) artificial soil matrix. About 30 000 Kalopanax pictus plants were propagated via somatic embryogenesis and grown to 3-yr-old plants. These results indicate that production of
woody medicinal Kalopanax pictus plantlets through somatic embryogenesis can be practically applicable for propagation. 相似文献
20.
Summary
Acmella radicans var. radicans propagation was established in vitro. This plant belongs to the Asteraceae from which some species are known for their insecticide, fungicide and antibacterial
activity. The complete Murashige and Skoog (MS) medium was the best in assisting seed germination. In order to obtain shoots
in vitro, a complete MS medium and half-strength MS medium were assayed with explants from leaves, nodes, and internodes. The best
medium for shoot production was the half-strength MS medium with no addition of plant growth regulators, and the highest shoot
propagation was from single-node explants. Regeneration of roots on shoot explants in the medium was obtained without the
addition of growth regulators. Of the plantlets that were acclimated, 90% of them were obtained from rooted shoots with completely
expanded leaves. The alkamide content was evaluated for each tissue and the higher concentration was observed in flower heads.
The main alkamides present in the leaves and the flower heads were N-(2-phenylethyl)-2Z,4E-octadienamide and 3-phenyl-N-(2-phenylethyl)-2-propenamide. This study describes the methodology for the establishment and propagation of Acmella radicans in vitro and the evaluation of different tissue alkamide contents in vitro and in the field. 相似文献