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1.
A reproducible protocol has been developed for high frequency plant regeneration from immature embryos of Argyrolobium roseum Jaub & Spach, an important medicinal legume. Green nodular calli were initiated from immature embryos excised from 10-d-old
pods in 70 % of cultures within 3 weeks when grown on Murashige and Skoog (MS) medium supplemented with 0.5 mg dm−3 benzylaminopurine (BAP) + 0.25 mg dm−3 indole-3-acetic acid (IAA). Subsequent transfer of 5 mm2 callus pieces to MS medium supplemented with BAP (0.5 mg dm−3) alone or in combination with IAA (0.25 mg dm−3) facilitated regeneration of multiple shoots. Organogenic calli bearing multiple shoots when transferred to MS medium supplemented
with BAP (0.5 mg dm−3) + IAA (0.25 mg dm−3) supported rapid shoot elongation. Shoot propagules subcultured to Gamborg's medium (B5) with 0.5 mg dm−3 indole-3-butyric acid (IBA) rooted with 80 % frequency and developed into phenotypically normal plants. Plantlets were successfully
acclimatized in a sterile mixture of sand and garden soil (1:1) under greenhouse and thereafter transferred to field beds. 相似文献
2.
Zephyr lily (Zephyranthes grandiflora), an important ornamental plant has been micropropagated in vitro after controlling microbial contamination by a pretreatment with 0.2 % Bavistin and 0.1 % Pantomycin for 4 h before final sterilization with 0.1 % mercuric chloride. In 67 % of the sterile cultures, 11 shoots on average were
regenerated directly from basal half of bulb scales in Murashige and Skoog (MS) medium containing 3 % sucrose and 2 mg dm−3 benzylaminopurine (BAP). Shoots emerged in bunches on a basal achlorophyllous bulbous part. Combination of 2 mg dm−3 BAP with 1 mg dm−3 gibberellic acid (GA3) enhanced shoot growth. Stout roots (maximum of 5–6 per shoot) were developed in presence of 1 mg dm−3 indole-3-butyric acid (IBA). Micro-bulbs showed potential of regeneration and could be used as secondary explants. The morphologically
identical plants derived by in vitro propagation were genetically identical as shown by PCR based ISSR marker analysis of genomic DNA. 相似文献
3.
An efficient and rapid plant regeneration system was established for zonal and scented geraniums using leaf discs as explants. Several explants, medium and culture conditions were studied to optimize shoot induction. Leaf discs taken from 4–5 weeks old in vitro grown plants, whatever the genotype, were more effective for shoot regeneration than those taken from greenhouse grown plants. Darkness proved to be a stimulating factor for shoot regeneration and the combination between NAA and two cytokinins gave the best results. Direct shoot regeneration (100%) was obtained from leaf discs of P. capitatum on half-strength MS medium supplemented with 0.5 mg l−1 NAA in combination with 1 mg l−1 of BAP and zeatin in darkness (11.4 shoots per explant). In the same medium and culture conditions, all P. graveolens leaf discs also exhibited direct shoot regeneration (7.3 shoots per explant). For P. x hortorum, 100% of leaf discs underwent shoot regeneration on a MS medium supplemented with 0.2 mg l−1 NAA in combination with 0.5 mg l−1 of BAP and zeatin in darkness (8.8 shoots per explant) or under low light conditions with 0.2 mg l−1 NAA and 1 mg l−1 of BAP and zeatin (7.5 shoots per explant). For this species, the best results for shoot elongation were obtained on half-strength MS medium gelled with Phytagel 0.3% (v/v). Whatever the genotype, all shoots rooted readily when transferred to diluted MS medium (MS/2) containing 1 mg l−1 IAA. Acclimatized plants grew normally and flowered in greenhouse conditions. Flow cytometry analysis made on leaves of acclimatized plants revealed that all the scented geranium plants are similar to mother plants while 71% of P. x hortorum plants which showed strong growth were tetraploid. 相似文献
4.
The object of study was the regeneration of Pharbitis nil by direct and indirect organogenesis. From fragments of roots, cotyledons, hypocotyls and epicotyls on Murashige and Skoog
nutrient solution (MS) supplemented with naphtalenacetic acid (NAA) or indolylacetic acid (IAA; both at 0.1 mg·dm−3 concentration) in the presence of benzylaminopurine (BAP), zeatin or kinetin (all at 5 mg·dm−3 concentration) only root organogenesis was obtained. Likewise, when using the two-step method (2 or 5 days exposure to NAA
or IAA at 2 mg·dm−3 concentration followed by exposure to BAP or zeatin at 1 or 2 mg·dm−3 concentration) root organogenesis was observed in all types of explants. Moreover, shoot buds were formed on fragments of
epicotyl exposed vertically in relation to the medium. However, attempts at regenerating complete plants from them failed,
so did the regeneration of P. nil from callus. The roots were formed in callus cultures only. 相似文献
5.
Plants of two cytotypes (2n=2x=20, and 2n=3x=30) of pinto peanut (Arachis pintoi Krapov. & W.C. Gregory) were regenerated through somatic embryogenesis. Embryogenic calli were induced from shoot tips or
immature leaves dissected from in vitro growing plants. In the case of the diploid peanut the best somatic embryogenesis was achieved when shoot tips were cultured
on Murashige and Skoog (MS) medium supplemented with 10 mg dm−3
Picloram (PIC) and 0.1 mg dm−3 6-benzylaminopurine (BAP) or when explants from immature leaves were cultured on MS + 10 mg dm−3 PIC + 0.01 mg dm−3 BAP. In the case of triploid peanut the highest number of somatic embryos was obtained when shoot tips were cultured on MS
+ 10 mg dm−3 PIC + 0.01 mg dm−3 BAP or when immature leaves were cultured on MS + 20 mg dm−3 PIC + 0.01 mg dm−3 BAP. Somatic embryos were converted into plants by culture on MS + 0.01 mg dm−3 naphthaleneacetic acid + 0.01 mg dm−3 BAP. Plants were successfully transferred to pots in greenhouse. 相似文献
6.
Vinayak Singh Namita Singh Chauhan Mohit Singh Asif Idris Raju Madanala Veena Pande Chandra Sekhar Mohanty 《Plant signaling & behavior》2014,9(10)
An in vitro method of multiple shoot induction and plant regeneration in Psophocarpus tetragonolobus (L.) DC was developed. Cotyledons, hypocotyls, epicotyls, internodal and young seedling leaves were used as explants. MS media supplemented with various concentrations of either thidiazuron (TDZ) or N6-benzylaminopurine (BAP) along with NAA or IAA combinations were used to determine their influence on multiple shoot induction. MS media supplemented with TDZ induced direct shoot regeneration when epicotyls and internodal segments were used as explants. TDZ at 3 mg L−1 induced highest rate (89.2 ± 3.28%) of regeneration with (13.4 ± 2.04) shoots per explant. MS media supplemented with BAP in combination with NAA or IAA induced callus mediated regeneration when cotyledons and hypocotyls were used as explants. BAP (2.5 mg L−1) and IAA (0.2 mg L−1) induced highest rate (100 ± 2.66%) of regeneration with (23.2 ± 2.66) shoots per explant. Mature plants produced from regenerated shoots were transferred successfully to the greenhouse. In a comparative study, the phenolics contents of various parts of greenhouse-grown plants with that of in vitro-raised plants showed significant variations. 相似文献
7.
Rapid shoot multiplication of Nyctanthes arbor-tristis L. was achieved from axillary meristems on Murashige and Skoog (MS) basal medium supplemented with 1.0–1.5 mg dm−3 6-benzylaminopurine (BA), 50 mg dm−3 adenine sulfate (Ads) and 3 % (m/v) sucrose. Inclusion of indole-3-acetic acid (IAA) in the culture medium along with BA
+ Ads promoted a higher rate of shoot multiplication. Maximum mean number of microshoots per explant (6.65) was achieved on
the MS medium supplemented with 1.5 mg dm−3 BA, 50 mg dm−3 Ads and 0.1 mg dm−3 IAA after 4 weeks of culture. The elongated shoots rooted within 13 to 14 d on half-strength MS medium supplemented with
either indole-3-butyric acid (IBA), IAA or 1-naphthaleneacetic acid (NAA) with 2 % sucrose. Maximum percentage of rooting
was obtained on medium having 0.25 mg dm−3 IBA and 0.1 mg dm−3 IAA. About 70 % of the rooted plantlets survived in the greenhouse. The in vitro raised plants were grown normally in the field. 相似文献
8.
This paper describes multiple shoot regeneration from leaf and nodal segments of a medicinally important herb Centella asiatica L. on Murashige and Skoog’s (MS) medium supplemented with a range of growth regulators. The highest number of multiple shoots
was observed on MS augmented with 3.0 mg dm−3 N6-benzylaminopurine (BAP) and 0.05 mg dm−3 α-naphthaleneacetic acid (NAA). Leaf explant showed maximum percentage of cultures regenerating shoots (81.6 %), with the
highest shoot number (8.3 shoots per explant) and the shoot length (2.1 cm) whereas, nodal explant showed less number of shoots
with callus formation at the base cut end. Successive shoot cultures were established by repeatedly sub-culturing the original
explant on a fresh medium. Rooting of in vitro raised shoots was best induced on half strength MS supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA) with highest percentage of shoot regenerating roots (76.8 %) with 3–4 roots per shoot. Plantlets
were acclimated in Vermi-compost and eventually established in soil. Contents of chlorophyll, total sugars, reducing sugars and proteins were estimated in
leaf tissue from both in vivo and in vitro raised plants. Chlorophyll content was higher in in vivo plants, whereas other three components were higher in in vitro plants. 相似文献
9.
High frequency of shoot formation was achieved from Solanum nigrum L. leaves on Murashige and Skoog (MS) medium without any callusing stage. Shoot forming ability was more pronounced on leaves
positioned dorsally. For shoot induction, 2.0 mg dm−3 benzylaminopurine and 1.5 mg dm−3 kinetin were observed to be the most effective plant growth regulators (PGRs). The present paper also describes first successful
induction of in vitro flowering in S. nigrum. The leaf derived shoots were excised and treated with various root promoting PGRs and 0.25 mg dm−3 indole-3-butyric acid produced maximum number of roots (15.2 per plant). Plants were later transplanted in field with 100
% survival. Solasodine content was higher in in vitro raised shoots and leaf derived callus, compared to ex vitro grown shoots. 相似文献
10.
The effect of auxin, GA and BAP on potato shoot growth and tuberization was investigated under in vitro condition. The shoot length of potato explants increased with the increasing of concentrations (0.5 – 10 mg dm−3) of IAA treatment especially with the addition of GA3 (0.5 mg dm−3), but was inhibited by BAP (5 mg dm−3). The root number and root fresh weight of potato explants increased with the increasing of IAA levels either in the presence
of GA3 (treatment IAA+GA) or not (IAA alone). However, no root was observed in the treatment IAA+BAP, instead there were brown swollen
calli formed around the basal cut surface of the explants. The addition of GA3 remarkably increased the fresh weight and diameter of calli. Microtubers were formed in the treatments of IAA+BAP and IAA
+ GA + BAP but not observed in the treatments of IAA alone or IAA + GA. IAA of higher concentrations (2.5 – 10 mg dm−3) was helpful to form sessile tubers. With the increasing of IAA levels, the fresh weight and diameter of microtubers increased
progressively. At 10 mg/L IAA, the fresh weight and diameter of microtubers in the treatment of IAA + GA + BAP were 409.6
% and 184.4 % of that in the treatment of IAA + BAP respectively, indicating the interaction effect of GA and IAA in potato
microtuberization. 相似文献
11.
Direct shoot regeneration from leaf explants of <Emphasis Type="Italic">Spilanthes acmella</Emphasis> 总被引:2,自引:0,他引:2
Multiple shoots of Spilanthes acmella Murr. were induced from nodal buds of in vivo and in vitro seedlings on Murashige and Skoog (MS) medium containing 1.0 mg dm−3 6-benzyladenine (BA) and 0.1 mg dm−3 α-naphthalene-acetic acid (NAA). Adventitious shoots were successfully regenerated from the leaf explants derived from the
above mentioned multiple shoots. The efficiency of shoot regeneration was tested in the MS medium containing BA, kinetin,
or 2-isopentenyl adenine in combination with NAA, indole-3-acetic acid (IAA), or indole-3-butyric acid (IBA) and gibberellic
acid. Maximum number of shoots per explant (20 ± 0.47) was recorded with 3.0 mg dm−3 BA and 1.0 mg dm−3 IAA. An anatomical study confirmed shoot regeneration via direct organogenesis. About 95 % of the in vitro shoots developed roots after transfer to half strength MS medium containing 1.0 mg dm−3 IBA. 95 % of the plantlets were successfully acclimatized and established in soil. The transplanted plantlets showed normal
flowering without any morphological variation. 相似文献
12.
A protocol was developed for plant regeneration of Melia azedarach L. by in vitro culture of apical meristem (0.5 mm in length). The influence of six clones was investigated. The culture procedure comprised
two sequential steps: 1) Induction of shoots by in vitro culture of axillary buds from adult trees (10–15 years old) by culture on Murashige and Skoog (1962) medium (MS) supplemented
with 0.5 mg·dm−3 BAP (6-benzylaminopurine), 0.1 mg·dm−3 IBA (indolebutyric acid), and 0.1 mg·dm−3 GA3 (gibberellic acid). The Multiplication of the regenerated shoots was achieved in MS + 0.5 mg·dm−3 BAP + 0.1 mg·dm−3 GA3. 2) In vitro culture of the apical meristems from the regenerated shoots in MS medium (0.7 %) supplemented with various combinations of
BAP and IBA.
Maximum shoot proliferation was obtained on MS medium supplemented with 0.5 mg·dm−3 BAP and 0.1 mg·dm−3 IBA.
Regenerated shoots were rooted on MS + 3.5 mg·dm−3 IBA (4 days) followed by subculture on MS lacking growth regulators (30 days). Complete plants were transferred to soil. 相似文献
13.
This report describes an efficient plant regeneration system for the medicinal plant Lythrum salicaria via direct adventitious shoot development from leaf and stem explants. Leaf explants were much more responsive to regeneration
than stem segments. Of the hormonal combinations tested, those involving thidiazuron (TDZ; 0.1, 0.3 or 0.5 mg dm−3) were more effective than the combinations of other hormones and 0.1 mg dm-3 TDZ combined with either indole-3-acetic acid
(IAA) or indole-3-butyric acid (IBA) was the most productive. Rooting was readily achieved when multiple shoots were singled
out and cultured on medium containing different auxins. IAA was the most effective on root development in terms of both the
number of roots per shoot and the frequency of rooted shoots. More than 90 % of the regenerants survived after hardening for
four weeks at gradually decreased air humidity. 相似文献
14.
The effect of various hormonal combinations on regeneration of shoots and roots from meristem-derived callus of Crocus sativus L. and activities of antioxidant enzymes have been studied. The most efficient regeneration occurred with 1.0 mg dm−3 1-naphthaleneacetic acid (NAA) + 1.0 mg dm−3 thidiazuron and 1.0 mg dm−3 NAA + 2.0 mg dm−3 kinetin. For sprouting, regenerated shoot were subcultured on Murashige and Skoog medium containing 1.0 mg dm−3 NAA + 1.0 mg dm−3 benzylaminopurine (BAP). Protein content and superoxide dismutase activity decreased in regenerated shoots and roots and
increased in sprouting shoots, while catalase (CAT), peroxidase (POX) and polyphenol oxidase (PPO) activities increased during
organogenesis and decreased in sprouting shoots. High CAT and PPO activities were detected in regenerated roots, whereas high
POX activity was observed in regenerated shoot. 相似文献
15.
Rapid micropropagation was achieved in Chlorophytum borivilianum Santapau and Fernandes using shoot base as explants. Multiple shoots were induced on Murashige and Skoog’s (MS) medium supplemented
with 3.0 mg dm−3 6-benzylaminopurine, 0.1 mg dm−3 1-naphthaleneacetic acid, 150 mg dm−3 adenine sulphates and 3 % saccharose. Rooting was readily achieved upon transferring the shoots onto half strength MS medium
supplemented with 0.1 mg dm−3 indolebutyric acid and 2 % saccharose. Micropropagated plantlets were hardened in the greenhouse and successfully established
in soil. Random amplified polymorphic DNA (RAPD) markers were used to evaluate the genetic stability of the micropropagated
plants. Thirty one arbitrary decamers were used to amplify genomic DNA from in vitro and in vivo plant material to assess the genetic stability. All RAPD profile analysis from micropropagated plants was genetically similar
to mother plants. 相似文献
16.
E. C. R. Tavano L. C. L. Stipp F. R. Muniz F. A. A. Mour?o Filho B. M. J. Mendes 《Biologia Plantarum》2009,53(2):395-399
In vitro organogenesis of Citrus volkameriana and C. aurantium was studied considering three explant types: epicotyl segment, internodal segment, and hypocotyl segment with attached cotyledon
fragment. The explants were cultured in medium according to Grosser and Gmitter (EME) supplemented with 0, 0.5, 1.0, 1.5,
and 2.0 mg dm− 3 6-benzyl-aminopurine (BAP), incubated firstly in darkness for 4 weeks, and then transferred to 16-h photoperiod for 2 weeks.
Comparing epicotyl and internodal segments, a higher percentage of responsive explants and a higher number of shoots per explant
were obtained with epicotyl segments, regardless of the BAP concentration. For C. volkameriana the highest percentage of responsive epicotyl segments (42 %) was obtained in EME with 1.0 mg dm−3 BAP, while for C. aurantium (59 %) in EME with 0.5 mg dm−3 BAP. The organogenesis efficiency was the best with the use of the hypocotyl segment with attached cotyledon fragment (77
% for C. volkameriana and to 75 % for C. aurantium). With this explant the morphogenesis occurred only in the hypocotyl region. The in vitro organogenesis was characterized by histological analyses showing that the morphogenic process started in the cambium region
near the explant cut end. 相似文献
17.
A rapid in vitro propagation of Holarrhena antidysenterica has been developed. Seedling cotyledonary nodes on Murashige and Skoog medium (MS) containing 2 mg dm−3 N6-benzyladenine (BA) produced highest number of multiple shoots. The shoot numbers were increased further upon subculture on
MS medium supplemented with 0.5 mg dm−3 BA. By repeated subculture of derived shoots, a high multiplication rate was established. The excised shoots were rooted
on MS basal medium without growth regulators. The in vitro formed shoots were also rooted ex vitro by dipping them in 2 mg dm−3 of indole-3-butyric acid (IBA) solution for 2 min before transferring them onto the hardening medium. Successful hardening
and further establishment (survival 90 %) of micropropagated plants under natural conditions was observed. 相似文献
18.
In the present study, in vitro regeneration system for a recalcitrant woody tree legume, Leucaena leucocephala (cvs. K-8, K-29, K-68 and K-850) from mature tree derived nodal explants as well as seedling derived cotyledonary node explants
was developed. Best shoot initiation and elongation was found on full-strength Murashige and Skoog (MS) medium supplemented
with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 100 mg dm−3 glutamine, 20.9 μM N
6-benzylamino-purine (BAP) and 5.37 μM 1-naphthalene acetic acid (NAA). Rooting was induced in half-strength MS medium containing
2 % (m/v) sucrose, 100 mg dm−3 myoinositol, 14.76 μM indole-3-butyric acid (IBA) and 0.23 μM kinetin. The cultivar K-29 gave the best response under in vitro conditions. Rooted plantlets were subjected to hardening and successfully transferred to greenhouse. Further, somatic embryogenesis
from nodal explants of cv. K-29 via an intermittent callus phase was also established. Pronounced callusing was observed on full-strength MS medium containing
3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 40.28 μM NAA and 12.24 μM BAP. These calli were transferred to induction medium and maximum number of globular
shaped somatic embryos was achieved in full-strength MS medium fortified with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 15.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 5.0 μM BAP and 1.0 mM proline. Moreover, an increase in endogenous
proline content up to 28th day of culture in induction medium was observed. These globular shaped somatic embryos matured in full-strength MS medium
with 3 % (m/v) sucrose, 100 mg dm−3 myoinositol, 10.0 μM BAP, 2.5 to 5.0 μM IBA and 0.5 mM spermidine. 相似文献
19.
A protocol for in vitro multiplication of Capparis decidua (Forsk.) Edgew. has been developed from cultured leaves procured from multiplying axillary shoots on the cultured nodal explants.
The highest efficiency of shoot formation was observed on Murashige and Skoog (MS) medium containing 2 mg dm−3 benzyladenine (BA) and 0.5 mg dm−3 1-naphthaleneacetic acid. The regenerated shoots were transferred to MS medium containing 3 mg dm−3 BA for growth and proliferation. Shoots above 2 cm in length were transferred to MS medium supplemented with 1 mg dm-3 indole-3-butyric
acid plus 0.5 mg dm−3 indole-3-acetic acid for root induction. No variation was detected among the micropropagated plants by randomly amplified
polymorphic DNA (RAPD) markers. 相似文献
20.
Rhododendron shoot regeneration was accomplished using either flower explants (each consisting of ovary with pedicel) of Rhododendron cvs. Nova Zembla and Irina or leaves isolated from in vitro grown Rhododendron catawbiense Michx. Multiple shoot tip clumps were obtained on Anderson's medium containing 0.5 to 1.5 mg dm−3 thidiazuron (TDZ) in combination with 12 to 15 mg dm−3 N6-[2-isopentenyl]adenine (2iP) and 1 to 3 mg dm−3 indole-3-butyric acid (IBA). After 16 weeks on the regeneration media, explants with shoot tip clumps were transferred for
shoot elongation to Anderson's medium with 3 mg dm−3 2iP. Two months later, the shoots have reached 5 to 40 mm in length and were fit for subcultivation.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献