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1.
Pretreatment in thidiazuron improves the in vitro shoot induction from leaves in Curculigo orchioides Gaertn., an endangered medicinal plant 总被引:1,自引:0,他引:1
T. Dennis Thomas 《Acta Physiologiae Plantarum》2007,29(5):455-461
Leaf regeneration via direct induction of adventitious shoots obtained from an endangered medicinal plant, Curculigo orchioides Gaertn. by pretreating with thidiazuron. C. orchioides is an endangered medicinal herb belonging to the family Hypoxidaceae. Direct inoculation of leaf pieces on MS medium supplemented
with various concentrations of BAP (2–8 μM) or TDZ (2–8 μM) alone or in combination with NAA (0.5 and 1.0 μM) produced low
shoot induction both in terms of % response and number of shoots per explant. Hence, leaf explants were pretreated with 15,
25 or 50 μM thidiazuron (TDZ), for 6, 24 or 48 h with the aim of improving shoot regeneration from cultured explants. After
pretreatment, explants were transferred to an agar solidified MS medium that was supplemented with BAP (4 μM), TDZ (6 μM),
BAP (4 μM) + NAA (1.0 μM), TDZ (6 μM) + NAA (0.5 μM). Control explants were incubated directly on the medium without any pretreatment.
The pretreatment of explants with 15 μM TDZ for 24 h significantly promoted the formation of adventitious shoots and the maximum
response was observed on MS medium supplemented with 6 μM TDZ. In this medium, 96 % cultures responded with an average number
of 16.2 adventitious shoots per explant. The percentage of leaf explants producing shoots and the average number of shoots
per explant were significantly improved when TDZ pretreated leaves were cultured onto MS medium supplemented with BAP or TDZ
alone or in combination with NAA. The rooted plantlets were successfully transplanted to soil with 90% success. The present
investigation indicated the stimulatory role of TDZ pretreatment in regulating shoot regeneration from leaf explants of C. orchioides. 相似文献
2.
Factors influencing reliable shoot regeneration from leaf explants of rapeseed (Brassica napus L.) were examined. Addition of AgNO3 to callus induction medium was significantly effective for shoot regeneration in all three genotypes initially tested. When
48 genotypes subsequently were surveyed, a large variation of shoot regenerability was observed, ranging from 100 to 0% in
frequency of bud formation and from 7.5 to 0 in the number of buds per explant. A significant correlation (r=0.84) was observed between the frequency of bud formation and the number of buds per explant. The shoot regenerability from
leaf explants was not related to that from cotyledonary explants (r=0.28). Histological observations showed that an organized structure developed from calluses produced at vascular bundle tissues
after 7 days of culture on callus induction medium, and they developed shoot apical meristems one week after transfer onto
shoot induction medium. Regenerated plantlets were obtained 2 months after the initiation of culture and they normally flowered
and set seeds. No alterations of morphology or DNA contents were observed in regenerated plants and their S1 progenies. 相似文献
3.
Maurizio Lambardi Kiran K. Sharma Trevor A. Thorpe 《In vitro cellular & developmental biology. Plant》1993,29(4):189-199
Summary Studies were undertaken to optimize tissue culture conditions for micropropagation of Aleppo pine (Pinus halepensis Mill.) from mature embryos and various explants of the embryo. Over 90% of the embryo explants gave rise to adventitious
buds within 4 wk. Intact embryos were the most suitable explants for shoot bud induction. Both isolated cotyledons and hypocotyls
produced adventitious buds, but these developed slowly and failed to elongate. N6-Benzyladenine (BA) alone at 5.0μM was the most effective cytokinin when added to gelled to gelled von Arnold and Eriksson’s (AE) medium containing 3% sucrose.
Adventitious bud development was achieved on hormone-free AE medium, and shoot elongation was optimum on three quarter-strength
Bornman’s MCM medium, with 0.1% conifer-derived activated charcoal. Shoots were multiplied on three-quarter strength MCM medium,
containing 5μM BA. To induce adventitious roots on the elongated shoots, pulse treatment with 1 mM IBA for 6 h, followed by the transfer of the shoots to sterile peat:vermiculite (1:1) mixture, was beneficial. After acclimatization
for 3 to 4 wk under mist, almost all the rooted shoots could be transplanted successfully to the greenhouse, where the plants
exhibited normal growth habit. Histologic studies on the ontogeny of adventitious shoot formation from mature embryo explants
revealed temporal structural changes in different parts of the explant. Induction of mitotic divisions on the shoot-forming
medium resulted in the formation of meristemoids in the epidermal and subepidermal layers of the explant, located initially
at both the tips of the cotyledons and the axils of adjacent cotyledons. Shoot buds arising in the axils of adjacent cotyledons
were due to new cell division and not to any preexisting meristem. 相似文献
4.
An efficient in vitro propagation system has been developed for rapid micropropagation of Soapnut (Sapindus trifoliatus Linn.), a medicinally and economically important tree from nodal (axillary bud) segments of seedlings. The frequency of shoot
regeneration from seedling node explant was influenced by the age of the seedlings, growth regulators and successive transfer
of the mother explant. Explants from 4-week-old seedlings yielded the maximum shoot regeneration frequency (97.22%) on full-strength
MS medium supplemented with 1.0 mg l−1 of 6-benzylaminopurine (BAP). After harvesting the newly formed shoots, the mother explants transferred to same medium subsequently
produced a maximum of 5.16 shoots per explant after third passage. Further improvement in the morphogenic response occurred
when the nodal explants excised from in vitro regenerated shoots were employed, and 6.89 shoots per explant were obtained
on the same medium after the third subculture. Optimal rooting (91.67%) was obtained by placing the micro-shoots in liquid
MS medium with 1.0 mg l−1 IBA for 24 h and then transferring to the agar solidified MS medium devoid of IBA. The micropropagated shoots with well-developed
roots were acclimatized and successfully transplanted to soil with 90% survival rate. Genetic stability of the regenerated
plants was assessed using random amplified polymorphic DNA (RAPD). The amplification products were monomorphic in micropropagated
plants and similar to those of mother plant. No polymorphism was detected revealing the genetic integrity of micropropagated
plants. This is the first report of an efficient protocol for regeneration of S. trifoliatus through organogenesis, which can be applied for further genetic transformation assays and pharmaceutical purposes. 相似文献
5.
The nodal and internodal explants excised from the orthotropic shoots of Sesbania sesban var. bicolor elicited the development of shoots directly from the explants as well as via an intervening callus phase on Nitsch (N) medium. On benzyladenine (BA) supplemented media, the adventitious shoot buds developed
involving a callus phase. An average of 8.9 ± 4.1 shoots developed per nodal explant on N medium containing 0.5 mg dm−3 BA in 95 % cultures, whereas 65 % cultures of internodal explants developed shoots with an average of 5.9 ± 3.6 shoots per
explant on N medium supplemented with 1.0 mg dm−3 BA. On kinetin (Kn) supplemented medium shoots developed directly from the surface of both the explants at all the concentrations
tried. Nodal explants on N medium supplemented with 1.5 mg dm−3 Kn developed an average of 12.5 ± 7.9 shoots per explant in 100 % cultures, while internodal explants induced an average
of 11.6 ± 7.4 shoots per explant in 75 % explants at 0.5 mg dm−3 Kn. The in vitro regenerated shoots developed roots when implanted on N medium supplemented with 2 mg dm−3 indole-3-butyric acid (IBA), after 30 d of inoculation. The in vitro developed plantlets were initially acclimatized under controlled conditions for four months, prior to their transfer to the
field.
This revised version was published online in July 2006 with corrections to the Cover Date. 相似文献
6.
In Pinus peuce zygotic embryo culture grown on Gresshoff and Doy (1972; GD) basal medium, 2.22 μM benzyladenine (BA) was superior in promoting
adventitious bud induction during 4 weeks comparing to kinetin or BA + kinetin. Shoot elongation was achieved on half-strength
GD medium devoid of plant growth regulators and containing activated charcoal. Pulse treatment with 1 mM indole-3-butyric
acid (IBA) for 2 h, followed by transfer to half-strength GD medium, produced the most efficient rooting. Rooted shoots were
transplanted to the greenhouse and plantlets continued to grow and developed into phenotypically normal plants. Up to 10 plants
per explant can be obtained within 36 weeks from culture initiation. 相似文献
7.
Hu Zhong Guo Guang-Qin Zhao Dong-Li Li Li-Hua Zheng Guo-Chang 《Russian Journal of Plant Physiology》2001,48(4):453-458
A method for fast plant regeneration via organogenesis directly from Lycium barbarumleaf explants has been developed. The key factor for shoot regeneration was the presence of benzyladenine (BA) in the medium. NAA could only induce root formation and explant callusing. Murashige and Skoog (MS) medium supplemented with 2 mg/l BA and 0.5 mg/l NAA is the most efficient condition for shoot formation, with up to 92.6% shoot regeneration and no callus formation. All adventitious shoots cultured on MS medium supplemented with 1 mg/l IAA formed an extensive root system. Regenerated plants were morphologically normal and were also proved to be diploid (2n = 24). Using the optimized regeneration system, the genetic transformation of L. barbarumwas carried out mediated by Agrobacterium tumefaciensEHA101(pIG121Hm). 11.8% leaf explants produced kanamycin-resistant shoots after infection by A. tumefaciens.The putative transgenic nature of plants was confirmed by GUS assay and PCR analysis. Expression of the nptIIgene in the regenerated plants was also detected by observing the callus formation by leaf pieces on MS medium containing 0.2 mg/l 2,4-D and 0–100 mg/l kanamycin. 相似文献
8.
S. Amutha M. Muruganantham A. Ganapathi 《In vitro cellular & developmental biology. Plant》2006,42(1):26-30
Summary Prolific shoot regeneration was achieved in mungbean Vigna radiata (L.) Wilczek from 3-d-old in vitro cotyledonary node and hypocotyl explants from seedlings derived from mature seeds on Murashige and Skoog (MS) medium supplemented
with thidiazuron (TDZ) (0.9 μM). An initial exposure to TDZ for 20 d and three successive transfers to fresh medium with reduced thidiazuron levels (0.09
μM) resulted in the regeneration of 104 shoots/explant from the cotyledon and 30 shoots/explant from the hypocotyl. Thidiazuron-associated
abnormalities such as short compact shoots, fasciation and leaf growth in the form of rosettes were observed in shoots regenerated
from hypocotyl explants. Both axillary and adventitious shoot formation from the explants were confirmed by histology. Through
repectitive cycles of regeneration in the presence of TDZ, the number of shoots that could be obtained from the two explant
classes within 80 d was significantly higher than with previous reports in mungbean 相似文献
9.
Many South African medicinal plants are over-collected for use in traditional medicines. This necessitates developing methods for increasing production. Micropropagation can be used as an alternative to conventional propagation methods. Twin-scales, cut from large parent bulbs, were cultured on MS medium (Murashige and Skoog, 1962) supplemented with 25 plant growth regulator combinations. Bulblets formed on twin-scales in 24 of the treatments. All explants formed bulblets on plant growth regulator-free medium. The effect of plant growth regulators, activated charcoal, explant orientation, explant origin and photoperiod on bulblet production was investigated. Bulblet formation was greatest when twin-scales were excised from the middle of the parent bulb, placed adaxial side down on plant growth regulator-free medium and kept in a 16 h photoperiod. 相似文献
10.
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
11.
An efficient protocol was developed for rapid clonal propagation of the important medicinal plant, Centella asiatica (L.) Urban, using shoot tip culture. High frequency bud break (88 %) and multiple shoot formation (16.8 shoots/shoot tip) were induced from a shoot tip segment, which was cultured on MS medium supplemented with 6‐benzylaminopurine (BAP) (17.76 μM) and gibberillic acid (GA3) (1.44 μM). Half‐strength Murashige and Skoog (MS) medium supplemented with naphthalen acetic acid (NAA) (10.74 μM) induced the maximum (27.66) number of roots. Plantlets with 3–4 fully expanded leaves and well‐developed roots were successfully transferred to potted soil which exhibited a 95 % survival. The protocol enables the harvest of more than 25,000 plantlets within 160 days starting from a single shoot tip explant. 相似文献
12.
Li Wei Gao Huan-Huan Lu Rong Guo Guang-Qing Zheng Guo-Chang 《Plant Cell, Tissue and Organ Culture》2002,71(3):259-262
Development of an efficient in vitro plantlet regeneration system from tubers of Stachys sieboldii (Miq.), a traditional Chinese medicinal plant and vegetable, is described. Adventitious shoots with an average of 9.1 (after 4 weeks of incubation) shoots per explant were obtained from 60.4% of tuber explants cultured on Murashige and Skoog (MS) medium containing 1.0 mg l–1 thidiazuron (TDZ), and after another 4 weeks of incubation on growth regulator free MS medium, 33.5 shoots per explant were achieved. Subcultures showed results that were similar to those of the initial culture. Excised shoots were rooted both on MS and on half strength MS medium with a frequency of about 95%. Excised shoots were also rooted ex vitro by directly planting them into cups containing sand. Regenerated plants with well developed shoots and roots were successfully transplanted to soil, after which they developed free of abnormalities. 相似文献
13.
N. S. Shekhawat T. S. Rathore R. P. Singh N. S. Deora S. R. Rao 《Plant Growth Regulation》1993,12(3):273-280
Genotype, age of tree, nature of explant and size (length and diameter), season of explant collection, explant position on medium, plant growth regulators and certain additives (ascorbic and citric acids, adenine sulphate, L-arginine, glutamine and ammonium citrate), incubation conditions, and subculturing period greatly influenced the in vitro clonal propagation of P. cineraria. The maximum number of 10–12 shoots were induced from the nodal shoot segment from pruned thorny adult trees on Murashige and Skoog's (MS) medium containing 0.1 mgl-1 indole- 3-acetic acid (IAA)+2.5 mgl-1 benzylaminopurine (BAP)+additives. Higher temperature (31+-2°C) and mixed (fluorescent and incandescent) light of 50 mol m-2 s-1 photon flux density for 12 h per day photoperiod favoured shoot induction and subsequent growth. Explants from thornless trees produced 6–8 shoots per explant on MS medium containing 0.1 mgl-1 IAA+5.0 mgl-1 BAP + additives. Nodal shoot segments obtained from root and stump sprouts produced multiple shoots. Root segments differentiated into multiple shoots on MS medium containing 0.5 mgl-1 indolebutyric acid (IBA)+2.5 mgl-1 BAP.Differentiated shoots multiplied best on MS medium containing 0.1 mgl-1 naphthalene acetic acid (NAA)+1.0 mgl-1 BAP + additives. To yield multiple shoots the original explant was transferred 6 times on fresh medium after harvesting the differentiated shoots. Shoots were rooted by pulsing with 100 mgl-1 IBA for 4 h and then culturing on hormone-free half strength MS medium. Initial dark incubation for 5 days at high temperature (33±2°C) was found essential for root induction from shoots which was 63% within two weeks. The rooted plantlets contained a consistent number of chromosomes (2n=28). It is suggested that the protocol developed could be useful for cloning of mature and tested trees of P. cineraria. 相似文献
14.
Summary The effect of indole-3-butyric acid (IBA) and 1-naphthaleneacetic acid (NAA) on lateral root formation was investigated in
adventitious root culture of Panax ginseng. Lateral root formation was affected by IBA (24.6 μM) or NAA (9.8 μM). Lateral root primordia emerged from the explant root pericycle after about 7 d of culture when the roots were cultured
on Schenk and Hildebrandt (SH) medium supplemented with 24.6 μM IBA or 9.8 μM NAA. However, no changes were observed in the explant root pericycle on auxin-free medium. The IBA treatment was more effective
for lateral root induction and root growth compared to NAA. In morphological and histological aspects, the lateral roots formed
under IBA treatment developed normally, while NAA-treated roots exhibited abnormal growth. The accumulation of total saponin
was greater in roots treated with IBA than with NAA. 相似文献
15.
Bodhipadma Kitti Noichinda Sompoch Padyencheun Winan Khunthacharoen Theerapong Chikhunthod Utorn Leung David W. M. 《Plant Cell, Tissue and Organ Culture》2011,105(3):465-469
The shoots developed from both the shoot tip and nodal explants of feathered amaranth (Celosia argentea var. plumosa—feathered cockscomb or plumed cockscomb) after 8 weeks of culture in the presence of either paclobutrazol or benzyladenine
(BA) were shorter than those developed on basal Murashige and Skoog (MS) medium (Physiol Plant, 15:473–497, 1962) alone. However, this retarding effect was more pronounced in the nodal explant culture. Shoot tip explants from 2-week-old
seedlings were more adversely affected by 0.85 or 1.7 μM paclobutrazol than those from older seedlings. In contrast, regardless
of preculture duration investigated nodal explants did not exhibit different response to three different concentrations of
paclobutazol. The response to 2.2 or 4.4 μM BA appeared to be largely independent of the age of the shoot tip explants or
preculture treatment of nodal explants. Shoots developed from nodal explants produced a higher number of terminal inflorescence
than those from shoot tip explants. Moreover, only lateral shoots from nodal explant culture formed inflorescence. Increased
preculture duration on basal MS medium could generally lessen the inhibitory effect of lower concentrations of paclobutazol
or BA on terminal or lateral inflorescence formation in nodal explant culture. 相似文献
16.
L. V. Hiregoudar H. N. Murthy J. G. Bhat A. Nayeem B. P. Hema E. J. Hahn K. Y. Paek 《Biologia Plantarum》2006,50(2):291-294
This report describes in vitro shoot induction and plant regeneration from mature nodal explants of Vitex trifolia L. on Murashige and Skoog (MS) medium fortified with benzylaminopurine (BAP), kinetin (KN), thidiazuron (TDZ), adenine (ADE),
and 2-isopentenyladenine (2-iP) (0.25 – 10.0 μM). Multiple shoots differentiated directly without callus mediation within
3 weeks when explants were cultured on medium supplemented with cytokinins. The maximum number of shoots (9 shoots per explant)
was developed on a medium supplemented with 5.0 μM BAP. Shoot cultures was established repeatedly subculturing the original
nodal explant on the same medium. Rooting of shoots was achieved on half strength MS medium supplemented with 0.5 μM naphthaleneacetic
acid (NAA). Rooted plantlets transferred to pots containing autoclaved soil and vermiculite mixture (1:1) showed 90 % survival
when transferred to outdoor. 相似文献
17.
In vitro culture of hypocotyl explants from Kandelia candel, a common mangrove species, on hormone-free Murashige and Skoog (MS) medium resulted in shoot formation. Since the hypocotyls showed good potential for in vitro shoot multiplication, the process of bud primordium formation was analyzed from a histological viewpoint. A wound periderm first appeared at the top, exposed cut surface of the explants. The wound-induced meristem continued to divide giving rise to suberized cells oriented towards the cut surface. After formation of the suberized cell layers, the meristem and its inner derivatives differentiated into multilayered, uniformly packed parenchyma cells. Bud primordia differentiated from the dense cytoplasmic cells of the wound-induced meristem just beneath the suberized layer near the severed vascular bundles. Each explant produced several visible shoot buds. Furthermore, histological sections revealed that additional bud primordia were present within the explant just underneath the suberized cells and that these bud primordia appeared to be arrested in their development. The fact that additional bud primordia were present within the explant suggests that further manipulation of the explant is helpful to maximize the potential of this system. 相似文献
18.
Summary A method of micropropagation through multiple shoot formation from axillary buds of mature tree and rootstock growths of Crataeva adansonii (DC.) Prodr. (a multipurpose tree) has been developed. Factors affecting multiplication rate included season, age of explant
source, explant type, type of bud, position of bud on the foliage twig, type of medium, various additives, and explant implantation
on the medium. The maximum number of buds was produced from the sixth to 10th axillary buds taken from foliage twigs of 40–50-d-old
rootstock growth in the months of October to December. At this time of the year the contamination was minimum. Optimum response
was recorded on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA; 3 mgl−1, 13.3μM) and 1-naphthaleneacetic acid (NAA; 0.05 mgl−1, 0.27 μM) after 21 d of culture. Shoot buds were further multiplied and maintained on BA (1–1.5 mgl−1, 4.4–6.7 μM) while shoots elongated on BA (0.1–0.5 mgl−1, 0.44–2.2 μM) supplemented medium. The number of shoots was further multiplied by using nodal segments of in vitro-regenerated shoots as microcuttings and repeated subculturing of stumps after excising the microshoots. In vitro rooting on growth regulator-free MS medium was possible with 70% of microshoots after 4 wk. From one nodal segment 150 plantlets
were produced within 14 wk. 相似文献
19.
White, filamentous, callus-like tissue was produced from the temperate marine brown alga Laminaria setchellii Silva under non-photosynthetic conditions. Explant disks from the medullary core of the stipe section were plated on solid medium (1.5 wt% agar in seawater base, pH 8) at 8 ± 2°C in the dark, and the formation of callus-like tissue was followed as a function of time up to 8 weeks. Although 30% of the total explants plated formed filamentous, callus-like growths, only 3 to 6% of the total explants produced relatively large amounts of filamentous and rounded callus-like cells. The fresh weight yield of this callus-like tissue was 3.74 ± 0.93 mg per 10 mm explant disk (0.053 g callus per g explant). Attempts at establishing a growing, cell suspension of the dispersed callus tissue in PES liquid medium at 24 μmol photon m?2 s?1 were not successful. 相似文献
20.
Somatic embryogenesis of Cyclamen persicum in liquid medium 总被引:1,自引:0,他引:1
Marc Kreuger Erik Postma Yvon Brouwer Gerrit-Jan van Holst 《Physiologia plantarum》1995,94(4):605-612
A method is described for the production of somatic embryos of Cyclamen persicum Mill. in liquid medium. Five steps are involved; initiation of embryogenic cell lines, proliferation of pro-embryogenic masses (PEMs) on auxin-containing medium, development of somatic embryos on hormone-free medium with high osmolarity, germination and subsequent plantlet formation. Cell lines were initiated by culturing the explant, the seedling tuber, directly in liquid medium. Three parameters were important for obtaining embryogenic cell lines; explant density, hormone concentrations and subculture regime. The rate of uptake of the hormones 2,4-dichlorophenoxyacetic acid (2,4-D) and kinetin influenced the formation of PEMs. Highly embryogenic cell lines were obtained only when PEMs had formed within 5–7 weeks. PEMs were proliferated for at least 24 months and could be isolated from each subculture for the production of somatic embryos. A high sucrose content (175 m M ) in the development medium without hormones ensured efficient embryo development from PEMs. A subsequent subculture in low sucrose concentration (58 m M ) induced the formation of a tuber, thus promoting germination. Arabinogalactan-proteins (AGPs) from carrot seeds and AGPs bound by the monoclonal antibody ZUM 18 increased the number of PEMs in a culture, showing that the activity of AGPs is not species specific. 相似文献