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1.
Murugesan Dhandapani Doo Hwan Kim Seung-Beom Hong 《In vitro cellular & developmental biology. Plant》2008,44(1):18-25
High-frequency plant regeneration of C. roseus cv. ‘little bright eye’ via somatic embryogenesis and organogenesis from five out of six explants was standardized. Two factors
were found to be important for regeneration: (1) the type of explants, and (2) the combination and concentrations of plant
growth regulators. The highest regeneration percentage through somatic embryogenesis was obtained from mature zygotic embryo
in MS medium supplemented with 7.5 μM of thidiazuron (TDZ). The mature embryo also regenerated efficiently via organogenesis
in MS medium supplemented with either 2.5 μM TDZ or 5.3 μM α-naphthalene acetic acid (NAA) and 2.2 μM 6-benzylaminopurine
(BA). Hypocotyl and cotyledon did not induce somatic embryogenesis and organogenesis in TDZ-containing medium but gave a maximum
percentage of shoots in MS medium supplemented with 5.3 μM NAA and 2.2 μM BA. Stem nodes and meristem tips showed better regeneration
via organogenesis in the medium supplemented with NAA and BA and in lower concentrations of TDZ. 相似文献
2.
Summary Tennessee coneflower [Echinacea tennesseensis (Beadle) Small] was regenerated from flower stalks, leaf sections from flowering plants, and hypocotyls and cotyledons from
seedlings. Murashige and Skoog medium (MS) supplemented with naphthaleneacetic acid (NAA) at 0.54 μM and thidiazuron (TDZ) at 22.7 μM yielded the most shoots per leaf explant. NAA and 6-benzylaminopurine concentrations for optimal shoot regeneration from
leaf, flower stalk, cotyledon and hypocotyl explants in MS media were 0.54 and 24.6μM, respectively. All explant types generated shoots; however, those derived from leaves and flower stalks produced the highest
number of shoots per explant and highest percentage of explants with shoots. Explants cultured on media containing high levels
of NAA (5.4–27 μM) formed calluses but no adventitious shoot. Leaf explants responded to a wider range of NAA concentrations than the other
explant types but shoots generated from flower stalks grew the fastest. While all cytokinins tested increased the number of
shoots per explant, the number of shoots in media containing TDZ was increased by nearly threefold. Regenerated shoots from
all explant types cultured on MS medium supplemented with 0.25 μM indole-3-butyric acid initiated roots within 4 wk; NAA was not effective for root induction. All vernalized plantlets developed
into plants that were morphologically identical to the source material. 相似文献
3.
Summary A procedure for plant regeneration, flower and plant formation from petiolar and inflorescence nodal explants of culantro
is discribed. Leaf petioles were excised from young leaves of non-flowering plants while nodal explants were excised from
the inflorescence. Explants were cultured in Murashige and Skoog (MS) medium alone or supplemented with 0.5μM naphthaleneacetic acid (NAA) and 0.9, 1.8, 4.5, or 9 μM thidiazuron (TDZ). All explants produced multiple shoots. In addition, nodal explants formed flowers. Shoot number, flower
number and shoot length were influenced by TDZ and NAA. Rooted shoots from both types of explants were transferred to soil
where plants were successfully established. 相似文献
4.
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
5.
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
6.
Summary An efficient in vitro propagation system was developed for Arnebia euchroma, an important Chinese traditional medicinal plant. The present study utilized thidiazuron (TDZ) for the induction of shoot
organogenesis on cotyledon and hypocotyl explants. The maximal number of shoots was obtained on the modified Linsmaier and
Skoog (LS) medium supplemented with 1.0 mgl−1 (4.5 μM) TDZ for 12d on cotyledon explants (8.6 shoots per cotyledon explant). Other cytokinins (kinetin and 6-benzyladenine) and
auxin (α-naphthaleneacetic acid) were not efficient in inducing regeneration on cotyledon explants. Browning of the basal
portion of the subcultured shoots could be significantly reduced when they were cultured on the modified LS medium supplemented
with 100 mgl−1 (33.3 μM) polyvinylpyrrolidone. Well-developed shoots formed roots on the same medium containing 1.0 mgl−1 (4.9 μM) indole-3-butyric acid. The efficient regeneration protocol reported here provides an important means of micropropagation
of this plant. Furthermore, this protocol is essential to future genetic improvement of plants via transformation protocols. 相似文献
7.
A high frequency adventitious shoot regeneration protocol was developed for henbane (Hyoscyamus niger L.) using thidiazuron (TDZ). Hypocotyl, cotyledon and stem explants were cultured on Murashige and Skoog (MS) medium supplemented with different concentrations of N6-benzylaminopurine and TDZ. MS medium supplemented with 16 μM TDZ was the most effective for providing 100 % regeneration frequency associated with a 19.53 shoots per hypocotyl explant. Plantlets were rooted on MS medium supplemented with different concentrations of indole-3-butyric acid (IBA) and α-naphthaleneacetic acid. High rooting and survival was achieved using half strength MS medium supplemented with 8 μM IBA.This study was supported by The State Planning Commission of Turkey (DPT) and University of Ankara (Project Nos.: 98K120640 and 2001K120240). 相似文献
8.
A rapid and efficient plant regeneration protocol for a wide range of alfalfa genotypes was developed via direct organogenesis. Through a successive excision of the newly developed apical and axillary shoots, a lot of adventitious
buds were directly induced from the cotyledonary nodes when hypocotyl of explants were vertically inserted into modified Murashige
and Skoog (MS) medium supplemented with 0.025 mg dm−3 thidiazuron (TDZ) and 3 mg dm−3 AgNO3. When the lower part of shoots excised from explants were immersed into the liquid medium with 1.0 mg dm−3 α-naphthaleneacetic acid (NAA) for 2 min, and then transferred to hormone free half-strength MS medium, over 83.3 % of the
shoots developed roots, and all plantlets could acclimatize and establish in soil. The protocol has been successfully applied
to eight genotypes, with regeneration frequencies ranging from 63.8 to 82.5 %. 相似文献
9.
Summary An in vitro culture procedure was established for repetitive embryogenesis and plant regeneration from seed-derived protocorms of Phalaenopsis amabilis var. formosa Shimadzu (Orchidaceae). Seed-derived protocorms were cultured on modified half-strength Murashige and Skoog (1962) basal medium (1/2MS) devoid
of plant growth regulators. After 45 d, 28.1% of protocorms formed embryos from their posterior regions. 1-Phenyl-3-(1,2,3-thiadiazol-5-yl)-urea
(TDZ; 0.45, 4.54, and 13.62 μM) promoted direct embryo formation. The best response was at 13.62 μM TDZ, and 100% of the protocorms formed a mean number of 13.5 embryos after 45d of culture. By contrast, naphthaleneacetic
acid (NAA) at 0.54 and 5.37 μM inhibited direct embryo formation. On basal medium devoid of plant growth regulators, 18.8% of primary proliferating embryos
could form more embryos. TDZ (0.45, 4.54, and 13.62 μM) also promoted this process. Proliferating embryos/protocorms were transferred to basal medium devoid of plant growth regulators
for plantlet formation. Plantlets were successfully obtained from the embryos after 4–6 wk. Following subculture every 6 wk
for three passages, the plantlets were transferred to sphagnum moss in a container for acclimatization in the greenhouse.
The survival rate was 100%. 相似文献
10.
Summary A method has been developed for the induction of adventitious shoots from leaf tissue of Echinacea pallida with subsequent whole-plant regeneration. Proliferating callus and shoot cultures were derived from leaf tissue explants
placed on Murashige and Skoog medium supplemented with 6-benzylaminopurine and naphthaleneacetic acid combinations. The optimum
shoot regeneration frequency (63%) and number of shoots per explant (2.3 shoots per explant) was achieved using media supplemented
with 26.6 μM 6-benzylaminopurine and 0.11 μM naphthaleneacetic acid. Rooting of regenerated shoot explants was successful on Murashige and Skoog medium, both with and
without the addition of indole-3-butyric acid. All plantlets survived acclimatization, producing phenotypically normal plants
in the greenhouse. This study demonstrates that leaf tissue of E. pallida is competent for adventitious shoot regeneration and establishes a useful method for the micropropagation of this important
medicinal plant. 相似文献
11.
The genus Tribulus is the source of a number of steroidal saponins and other bioactive compounds which are of medicinal and pharmaceutical importance and plant regeneration of Tribulus terrestris has been reported. The objective of this study was to evaluate the potential of immature zygotic embryos of Tribulus terrestris as an explant for plant regeneration. Embryos were cultured on MS medium supplemented with 1-naphthaleneacetic acid (NAA), 2,4-dichlorophenoxyacetic acid (2,4-D) and thidiazuron (TDZ), alone or in combination and callus and shoot or embryo formation evaluated. With 2.5 mg/l NAA or 2,4-D, callus formation frequency was 100% but 57% with 2.5 mg/l TDZ. The combination of 2.5 mg/l TDZ and NAA or 2,4-D also elicited callus formation frequency of 100%. The callus formation frequency was lower with lower levels of these growth regulators. On a medium with 0.5 mg/l TDZ, 17.4% of the 2,4-D-derived callus (2.5 mg/l), developed embryo-like structures and this increased to 37.3 and 41.4% respectively, when TDZ was combined with 0.5 mg/l indole-3-butyric acid (IBA) or 2,4-D. Both shoot formation and embryo-like structures developed in cultures with 2.5 mg/l TDZ, alone or in combination with 0.5 mg/l IBA or 2,4-D. The optimum sucrose level for morphogenetic response of embryo-derived callus was between 5.0 and 7.5%. Embryo-like structures were also observed when the 2,4-D-derived callus was cultured in a liquid containing benzyladenine (BA) and IBA. Plants were regenerated from both embryo-like structures and shoot buds on solid MS medium containing 0.2 mg/l IBA and rooted plantlets were transferred to soil. 相似文献
12.
Somatic embryos regenerated in high-frequency (up to 100 %) on immature cotyledons of Azadirachta indica at a low concentration of thidiazuron (TDZ; 0.5 M). Regeneration occurred exclusively from abaxial surface. Frequency of regeneration declined with the age of the cotyledons: on semimature cotyledons regeneration occurred only from some regions of the abaxial surface and on mature cotyledons it was confined to corners. However, an increase in concentration of TDZ to 1.0 M improved the regeneration response. Higher regenerative capacity of immature cotyledons was due to presence of milk in immature fruits because regeneration response of immature cotyledons declined on washing of cotyledons for 24 h in liquid medium, and milk from immature fruits augmented the regeneration response of mature cotyledons. Somatic embryos regenerated readily on hormone-free medium and plantlets derived were able to survive after transfer to soil. 相似文献
13.
P. Baskaran P. Velayutham N. Jayabalan 《In vitro cellular & developmental biology. Plant》2009,45(4):407-413
An effective protocol was developed for in vitro regeneration of the Melothria maderaspatana via indirect organogenesis in liquid and solid culture systems. Organogenesis was achieved from liquid culture calluses derived from leaf and petiole explants of mature plants. Organogenic calluses (98.2?±?0.36 and 94.8?±?0.71%) were induced from both leaf and petiole explants on Murashige and Skoog (MS) liquid medium containing 6.0 µM 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.5 µM thidiazuron (TDZ); and 6.0 µM 2,4-D and 1.0 µM benzyladenine (BA) combinations, respectively. Adventitious shoot regeneration (68.2?±?0.06 shoots per explant) was achieved on MS medium supplemented with 2.0 µM BA, 4.0 µM TDZ, 10% v/v coconut water and 0.06 mM glutamine from leaf-derived calluses. Petiole-derived calluses produced adventitious shoots (45.4?±?0.09 shoots per explant) on MS medium fortified with 2.0 µM BA, 4.0 µM TDZ, 10% v/v coconut water, and 0.08 mM glutamine. Elongation of shoots occurred in MS medium with 2.0 µM gibberellic acid (GA3). Regenerated shoots (2–3 cm in length) rooted (74.2?±?0.38%) and hardened (85?±?1.24%) when they were transferred to 1/2-MS medium supplemented with 3.0 µM indole-3-butyric acid (IBA) followed by garden soil, vermiculate, and sand (2:1:1 ratio) mixture. The elongated shoots (4–5 cm in length) were exposed simultaneously for rooting as well as hardening (100%) in moistened [(1/8-MS basal salt solution with 5 µM IBA and 100 mg l?1 Bavistin® (BVN)] garden soil, vermiculate, and sand (2:1:1 ratio) mixture. Subsequently, the plants were successfully established in the field. The survival percentage differed with seasonal variations. 相似文献
14.
Gunaratnam Thirukkumaran Valentine Otang Ntui Raham Sher Khan Masahiro Mii 《Plant Cell, Tissue and Organ Culture》2009,99(1):109-115
Efficient shoot regeneration and Agrobacterium-mediated genetic transformation systems were developed for Petunia hybrida cv. Mitchell. Leaf explants of petunia were cultured on Murashige and Skoog (MS) medium with different concentrations of thidiazuron (TDZ) without auxin. The highest frequency of shoot regeneration (52.1%) and mean number of shoots per explant (4.1) were obtained on medium containing 2 mg l?1 TDZ. Leaf explants inoculated with Agrobacterium tumefaciens strain EHA101/pIG121Hm harboring ß-glucuronidase (uidA) and hygromycin resistance genes developed putative transformant shoots. The highest frequency of shoot regeneration (22.5%) and mean number of transformant shoots per explant (2.4) were obtained on a selection medium consisting of the above described regeneration medium and containing 25 mg l?1 hygromycin as the selection agent. Approximately 95% of putative transformant shoots expressed the uidA gene following histochemical ß-glucuronidase (GUS) assay. These were confirmed to be transgenic by PCR analysis and Southern blot hybridization. 相似文献
15.
Hai Ping Hong Hongyi Zhang Paula Olhoft Steve Hill Hunt Wiley Effie Toren Helke Hillebrand Todd Jones Ming Cheng 《In vitro cellular & developmental biology. Plant》2007,43(6):558-568
A regeneration and transformation system has been developed using organogenic calluses derived from soybean axillary nodes
as the starting explants. Leaf-node or cotyledonary-node explants were prepared from 7 to 8-d-old seedlings. Callus was induced
on medium containing either Murashige and Skoog (MS) salts or modified Finer and Nagasawa (FNL) salts and B5 vitamins with
various concentrations of benzylamino purine (BA) and thidiazuron (TDZ). The combination of BA and TDZ had a synergistic effect
on callus induction. Shoot differentiation from the callus occurred once the callus was transferred to medium containing a
low concentration of BA. Subsequently, shoots were elongated on medium containing indole-3-acetic acid (IAA), zeatin riboside,
and gibberellic acid (GA). Plant regeneration from callus occurred 90 ∼ 120 d after the callus was cultured on shoot induction
medium. Both the primary callus and the proliferated callus were used as explants for Agrobacterium-mediated transformation. The calluses were inoculated with A. tumefaciens harboring a binary vector with the bar gene as the selectable marker gene and the gusINT gene for GUS expression. Usually 60–100% of the callus showed transient GUS expression 5 d after inoculation. Infected
calluses were then selected on media amended with various concentrations of glufosinate. Transgenic soybean plants have been
regenerated and established in the greenhouse. GUS expression was exhibited in various tissues and plant organs, including
leaf, stem, and roots. Southern and T1 plant segregation analysis of transgenic events showed that transgenes were integrated into the soybean genome with a copy
number ranging from 1–5 copies. 相似文献
16.
The organogenic competence of leaf explants of eleven Carthamus species including C. tinctorius on Murashige and Skoog (MS) medium supplemented with different concentrations of thidiazuron (TDZ) + α-naphthaleneacetic
acid (NAA) and 6-benzyladenine (BA) + NAA was investigated. Highly prolific adventitious shoot regeneration was observed in
C. tinctorius and C. arborescens on both growth regulator combinations and the shoot regeneration frequency was higher on medium supplemented with TDZ + NAA.
Nodal culture of nine Carthamus species on media supplemented with BA and kinetin (KIN) individually revealed the superiority of media supplemented with
BA over that of KIN in facilitating a higher shoot proliferation index. Proliferating shoots from axillary buds and leaf explants
were transferred to medium supplemented with 1.0 mg dm−3 KIN or 0.5 mg dm−3 BA for shoot elongation. Elongated shoots were rooted on half-strength MS medium supplemented with 1.0 mg dm−3 each of indole-butyric acid (IBA) and phloroglucinol. The plantlets thus obtained were hardened and transferred to soil. 相似文献
17.
A system for in vitro regeneration of Aloe arborescens was developed using young inflorescences as explants. Different phytohormone combinations of N-phenyl-N′-1,2,3-thiadiazol-5-yl urea (TDZ), benzyladenine (BA), 6-(γ,γ-dimethylallyl-amino)purine riboside (2iPR), zeatin ribozide (ZR), N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU) and kinetin (K), with or without ancymidol, were examined in order to induce plant regeneration. Efficient shoot regeneration was initiated on Murashige and Skoog (MS) medium supplemented with BA or TDZ. MS medium enriched with 19.6, 22.2 μM BA and 3.92 μM ancymidol (MSBA5/1 medium), promoted organogenesis enabling 87.3% of the explants to regenerate 6.04 ± 1.79 shoots/explant. Subsequent shoot elongation and plant regeneration were strongly affected by the medium composition used for shoot induction. Optimal elongation (three to four shoots per explant) was obtained when shoots, initiated on MSBA5/1 medium, were subsequently transferred onto MS containing only 4.4 μM BA. Rooting was performed on MS media lacking growth regulators. Histological analysis revealed that the initiated shoots originated from the receptacle tissue surrounding the residual vascular tissue of the flower buds. 相似文献
18.
Direct shoot regeneration was achieved from immature inflorescence explants of Chlorophytum arundinaceum and C. borivilianum on half-strength Murashige & Skoog (MS) medium supplemented with 3.0 mg L−1 BA, 150 mg L−1 Ads, 0.1 mg L−1 NAA and 3% (w/v) sucrose under a 16-h photoperiod. The shoot buds developed within 2–3 weeks of culture. High frequency of
shoot bud regeneration was achieved when cultured on similar medium in subsequent subcultures. The apex portion (Type I) of
the inflorescence produced more shoot buds as compared to the middle ones (type II). More than 75% of the terminal segment
explants produced shoot buds within 4-week of culture. Response of basal portion (Type III) was negative for shoot bud initiation.
Shoots rooted on half-strength basal MS medium supplemented with half-strength MS medium, 0.1 mg L−1 IAA and 2% (w/v) sucrose. Micropropagated plantlets were hardened in the green house and successfully established in the
soil where 90% of the plants survived. This protocol would be useful for commercial micropropagation and genetic improvement
prograrmme. 相似文献
19.
The attempts of this investigation were to develop a system for plant regeneration from explants of adult plants and its use for genetic transformation of important commercial Pelargonium zonale hybrid and P. peltatum hybrid cultivars. To this aim, leaf blade and petiole explants of eight cultivars were cultured on modified MS (Murashige and Skoog, 1962) medium with two concentrations of TDZ, BA, and zeatin (5 and 20 M). Petiole explants showed a higher regeneration response than leaf blade explants and TDZ was the most effective cytokinin. Petioles of 16 cultivars were incubated on medium containing 5, 10, 15, and 20 M TDZ, respectively, in order to identify the most effective TDZ concentration. For the majority of genotypes 10 M TDZ resulted in the best regeneration response. Cefotaxim at 500 mg l –1 had no effect on shoot regeneration and did not show interaction with glufosinate. For selection of transgenic cells, a concentration of 2.5 M glufosinate was shown to be appropriate. LBA4404 and EHA101 Agrobacterium strains carrying pIBGUS vector with pat gene as selectable marker gene and GUS gene as reporter gene were compared in transformation studies. With regard to GUS expression in petiole explants 16 days after coculture, better results were obtained with EHA 101 than with LBA 4404. 相似文献
20.
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