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1.
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
2.
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
3.
Naomita V. Dalal Ravishankar V. Rai 《In vitro cellular & developmental biology. Plant》2001,37(6):820-823
Summary Biotechnology has offered a nonconventional method of plant propagation and has been intensively applied as a conservation
strategy for sustaining biodiversity for rare plants. In vitro conservation through micropropagation of Ochreinauclea missionis, a rare, endemic and medicinal tree species of Western Ghats in Karnataka region of India is reported. Multiple shoots were
initiated from nodal explants on Murashige and Skoog (MS) medium supplemented with 8.8 μM 6-benzylaminopurine (BA) and 0.3% (w/v) activated charcoal. Shoots were elongated in MS medium with a combination of 2.2
μM BA and 5.3 μM α-naphthaleneacetic acid (NAA) or growth regulator-free medium. Individual shoots with a minimum of one node were excised
and rooted in vitro on MS medium with 0.3% activated charcoal or ex vitro rooted by treatment with 49 μM indole-3-butyric acid (IBA) for 30 min. Regenerants acclimated in Soil-rite exhibited 65% survival in the greenhouse. 相似文献
4.
Sancak Cengiz Mirici Semra Özcan Sebahattin 《Plant Cell, Tissue and Organ Culture》2000,61(3):231-235
An in vitro propagation protocol has been developed from mature trees of Pittosporum napaulensis. The best bud proliferation (83.1%), shoot number (21 axillary shoots/ explant) and shoot length (5.5 cm) was achieved in
Murashige and Skoog (MS) medium supplemented with 5.0 μM N−6 benzyladenine and 0.1 μM α- naphthalene acetic acid. Of the three cytokinins tested (N−6 benzyladenine, kinetin and thidiazuron), N−6 benzyladenine proved to be the best for shoot induction. Shoot regeneration potential varied among genotypes. Regenerated
shoots rooted after 48 hours treatment on half-strength MS liquid medium supplemented with 20 μM indole-3-butyric acid. Rooted
shoots transferred to 120 g (w/v) soilrite + sand + soil (1:1:1) mixture showed 70% survival. Twenty-one plantlets are growing
well in green house conditions.
This revised version was published online in August 2006 with corrections to the Cover Date. 相似文献
5.
C. L. Huang M. T. Hsieh W. C. Hsieh A. P. Sagare H. S. Tsay 《In vitro cellular & developmental biology. Plant》2000,36(3):220-224
Summary Rapid in vitro propagation of Limonium wrightii (Hance) Ktze. (Plumbaginaceae), an endangered medicinal plant, was achieved by culturing the shoot-tip (primary and lateral),
leaf- and influorescence-node explants. MS (Murashige and Skoog, 1962) medium supplemented with 8.87 μMN6-benyladenine (BA) and 1.07 μM α-naphthaleneacetic acid (NAA) supported induction of adventitious shoots from the shoot-tip, inflorescence-node and middle
and basal parts of leaf explants after 60 d of culture. Adventitious shoots were multiplied by subculturing on MS medium supplemented
with BA (2,21–17.75 μM) in combination with NAA (1.07 μM). The percentage of explants forming shoots and the average number of adventitious shoot buds produced per explant were stimulated
by increasing the strength (1/4x, 1/2x, 1x, 2x) of the MS medium. Shoots were rooted on MS basal medium with 4.92 μM indole-3-butyric acid. Plantlets with a morphologically normal appearance produced from adventitious shoots were transferred
to soil and acclimated in the growth chamber for 1 mo. 相似文献
6.
Summary
In vitro methods were applied to the only remaining plant of the Meelup Mallee (Eucalyptus phylacis), a critically endangered species from the southwest of Western Australia. Shoot explants were initiated into culture using
a 1/2 MS [Murashige and Skoog basal medium (BM) for all experiments] liquid medium supplemented with 1% (w/v) activated charcoal,
which was replenished twice daily, followed by transfer of explants to agar medium supplemented with 0.5 μM zeatin. Explants were cultured under low intensity lighting (PPFD of 5–10 μmol m−2s−1) to minimize blackening of tissues, and some explants were induced to produce nodular green calluses in response to BM supplemented
with 5 μM thidiazuron. Nodular green calluses were induced to form adventitious shoots following transfer to medium supplemented with
0.5 μM zeatin and 1 μM gibberellic acid, A4 isomer (GA4). Development of shoots was completed on 1 μM zeatin + 0.1 μM 6-benzylaminopurine (BA) in vented culture tubes. Regenerated shoots were sequentially cultured on medium containing 0.5
μM zeatin + 0.2 μM indoleacetic acid (IAA) followed by either 0.5 μM zeatin + 1μM GA4 for shoot elongation or 1 μM zeatin + 0.5 μM IAA to optimize shoot growth. Rooted microshoots were produced after 4 weeks on 5 μM indolebutyric acid (IBA) and survived acclimatization and transfer to potting mixture. 相似文献
7.
Summary An efficient protocol was established for in vitro shoot multiplication from nodal explants of Clitoria ternatea on semisolid Murashige and Skoog (MS) basal medium supplemented with 8.9μM 6-benzylaminopurine (BA). Inclusion of 1-naphthaleneacetic acid (NAA) in the culture medium along with BA promoted higher
rates of shoot multiplication than BA alone. The rate of shoot multiplication was maximum (5.21) after 4 wk of culture on
MS basal medium supplemented with 8.9μM BA and 1.34μM NAA. The elongated shoots rooted within 7–8d in half-strength MS basal salts supplemented with 1.34μM NAA and 2% (w/v) sucrose. About 85% of the rooted plantlets were acclimatized and transferred to the greenhouse. 相似文献
8.
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
9.
Summary A protocol has been developed for in vitro plant regeneration from cotyledonary nodes of Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes derived from 20-d-old axenic seedlings grown on Murashige and
Skoog (MS) medium containing 2.22–13.32 μM benzyladenine (BA) or 2.32–13.93 μM kinetin alone or in combination with 0.26 μM α-naphthaleneacetic acid (NAA). The highest frequency of responding explants (85%) and maximum number of shoots per explant
(9.5) were obtained on MS medium supplemented with 4.44 μM BA and 0.26 μM NAA after 15 wk of culture. A proliferating shoot culture was established by repeatedly subculturing the orginal cotyledonary
nodal explant on fresh medium after each harvest of the newly formed shoots. Nearly 30% of the shoots formed roots after being
transferred to half-strength MS medium containing 9.84 μM indole-3-butyric acid after 25 d of culture. Fifty percent of shoots were also directly rooted as microcuttings on peat moss,
soil, and compost mixture (1∶1∶1). About 52% plantlets rooted under ex vitro conditions were successfully acclimatized and established in pots. 相似文献
10.
A. V. Raghu S. P. Geetha Gerald Martin Indira Balachandran P. N. Ravindran 《In vitro cellular & developmental biology. Plant》2006,42(6):584-588
Summary A protocol was developed for rapid clonal propagation of the important medicinal climber, Tinospora cordifolia, through in vitro culture of mature nodal explants. Shoots were initiated on both Murashige and Skoog (MS) medium and woody plant medium (WPM)
supplemented with 2.32 μM kinetin (KIN). Of the two basal media tested, WPM was found to be superior to MS medium for the induction of multiple shoots.
Among the cytokinins tested, N6-benzyladenine (BA) was more effective than KIN for axillary shoot proliferation. KIN was superior to BA in terms of shoot
elongation. An average multiplication rate of 6.3 shoots per explant was obtained with WPM supplemented with 8.87 μM BA. Shoot clumps harvested from this medium were transferred to WPM supplemented with 2.22 μM BA and 4.65 μM KIN for shoot elongation. Elongated shoots were rooted in half-strength MS medium supplemented with 2.85 μM indole-3-acetic acid (IAA). Rooted plantlets were successfully transferred to sand and established with 80% survival. 相似文献
11.
V. Ravishankar Rai 《In vitro cellular & developmental biology. Plant》2002,38(4):347-351
Summary
Nothapodytes foetida (Wight) is a small evergreen tree and the extract from this tree is used to make the antileukaemia and antitumoral compound
camptothecin. Due to exploitation of this resource, efficient methods for rapid propagation of N. foetida are highly desirable. Multiple shoots were induced on hypocotyl segments of 20–25-d-old seedlings of N. foetida cultured on Murashige and Skoog (MS) medium supplemented with different concentrations and combinations of cytokinins. The
highest shoot multiplication was achieved on MS medium containing thidiazuron (TDZ) at the concentration of 2.2 μM. Inhibition of shoot elongation by TDZ was overcome by transferring shoot cultures to medium containing 2.2 μM benzylaminopurine which produced healthy shoots after three additional subcultures. The production of shoots was further
promoted by repeated subculturing of original explants on fresh multiplication medium after each harvesting of the newly formed
shoots. In vitro rooting was best induced (87%) in shoots excised from proliferated shoot cultures on one-fourth MS medium augmented with
5.7 μM indole-3-acetic acid and 2.4 μM indolebutryic acid (IBA). In vitro-developed shoots were also rooted ex vitro by dipping in 49 μM IBA for 10 min. In vitro- and ex vitro-rooted plants were successfully acclimatized and established in greenhouse conditions. 相似文献
12.
Summary Axillary and terminal buds from suckers of Ananas comosus cv. Phuket were established on Murashige and Tucker-based (MT) medium with 2.0 mgl−1 (9.8 μM) indolebutyric acid, 2.0 mgl−1 (10.74 μM) naphthaleneacetic acid, and 2.0 mgl−1 (9.29 μM) kinetin, followed by multiplication on Murashige and Skoog-based (MS) medium containing 2.0 mgl−1 (8.87 μM) benzyladenine (BA) to provide a continuous supply of axenic shoots. Leaves, excised from such cultured shoots, produced
adventitious shoots from their bases when these explants were cultured on MS medium containing 0.5 mgl−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid (2,4-D) and 2.0 mgl−1 (8.87 μM) BA. Embryogenic callus was produced when leaf explants were cultured on MS medium with 3.0 mgl−1 (12.42 μM) 4-amino-3,5,6-trichloropicolinic acid (picloram). Somatic embryos developed into shoots following transfer of embryogenic
tissues to MS medium with 1.0 mgl−1 (4.44 μM) BA. Cell suspensions, initiated by transfer of embryogenic callus to liquid MS medium with 1.0 mgl−1 (4.14 μM) picloram or 1.0 mgl−1 (4.52 μM) 2,4-D, also regenerated shoots by somatic embryogenesis, on transfer of cells to semisolid MS medium with 1.0 mgl−1 (4.44 μM) BA. All regenerated shoots rooted on growth regulator-free MS medium, prior to ex vitro acclimation and transfer to the glasshouse. These studies provide a baseline for propagation, conservation, and genetic manipulation
of elite pineapple germplasms. 相似文献
13.
Summary An efficient in vitro plant regeneration system from cotyledons was established in tetraploid Isatis indigotica Fort. Factors influencing shoot regeneration from cotyledons, including culture medium type, combinations of plant growth
regulators, and sucrose concentrations in the medium, as well as illumination were investigated. Murashige and Skoog's (MS)
medium was found to be best for promoting shoot regeneration, followed by Gamborg's B5 and White's medium. The highest shoot regeneration frequency was achieved from cotyledons cultured on MS medium supplemented
with 2.0 mgl−1 (8.9 μM) 6-benzyladenine and 1.0 mgl−1 (5.4 μM) α-naphthaleneacetic acid (NAA), with 97.9% regeneration, associated with a high number of multiple shoots developed per
explant (8.6 shoots per explant). A sucrose concentration of 3% present in the medium and light conditions were beneficial
for shoot regeneration. The shoots developed were rooted in a half-strength MS medium supplemented with 1.0 mgl−1 (5.4 μM) NAA and successfully transplanted in soil in pots with over 85% survival. The establishment of an efficient plant regeneration
procedure from cotyledons provides a basis for the rapid in vitro multiplication of tetraploid Isatis indigotica Fort., one of the most extensively used medicinal plants in China currently under great shortage. 相似文献
14.
Cao Dinh Hung Krystyna Johnson Fraser Torpy 《In vitro cellular & developmental biology. Plant》2006,42(6):548-552
Summary An efficient protocol for in vitro propagation of the valuable medicinal plant, Wasabia japonica (Miq.) Matsumura is described through shoot tip proliferation and direct regeneration. Multiple shoots were induced from
shoort tips cultured on Murashige and Skoog (MS) semi-solid medium containing various concentrations (0.5–50 μM) of N6-benzyladenine (BA), thidiazuron, kinetin, and zeatin. A comparison was made on shoot multiplication between semi-solid and
liquid culture media. Well-developed shoots were obtained using full-strength MS semi-solid medium containing 5.0 μM BA. However, the greatest shoot proliferation was achieved on either full- or half-strength MS liquid media supplemented
with 5.0 μM BA for 4 wk (15.3±0.9 and 15.0±0.7 shoots per explant, respectively), and on half-strength MS liquid medium for 6 wk (25.8±1.3
shoots per explant) in culture. In contrast, the maximum number of shoots per explant on full-strength MS semi-solid medium
was achieved with either 5.0 μM BA (10.4±0.6 shoots per explant) or 10.0 μM kinetin (10.9±0.8 shoots per explant). Fresh weight of explants and length of shoots derived from full-strength MS liquid
medium (1055±77 mg and 34.2±1.0 mm, respectively) were significantly higher than those derived from full-strength MS semisolid
medium (437.6±17.3 mg and 15.4±0.7 mm, respectively). Quarter-strength MS liquid medium had no significant difference in shoot
proliferation when compared to quarter-strength MS semi-solid medium. Elongated shoots were separated and rooted on half-strength
MS semi-solid media fortified with 1-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), or indole-3-acetic acid (IAA)
ranging from 0.1 to 10.0 μM. Root formation was greatest with IBA when compared with IAA and NAA. One hundred percent of shoots were rooted on half-strength
MS medium with 5.0 μM IBA, while vigorous roots were obtained with 10.0 μM IBA. Micropropagated plantlets were successfully established in soil with 95% survival rate after heardening. 相似文献
15.
An improved micropropagation protocol has been developed for teak (Tectona grandis). Nodal explants placed on MS medium supplemented with 22.2 M benzylaminopurine and then serially transferred to fresh medium after 12, 24, 48 and 72 h gave maximum culture establishment (76.8%). Establishment was reduced when explants were retained in the initial culture medium longer than 12 h. Explants collected in May showed maximum (76.8%) response. Placement of the explants on MS medium supplemented with 22.2 M benzylaminopurine and 0.57 M indole-3-acetic acid resulted in the maximum average number of shoots. In vitro raised micro shoots were rooted ex vitro by dipping in indole-3-butyric acid (9.8 mM) for 2 min followed by planting in polyethylene pots containing a soil:vermiculite (1:1 v/v) mixture. This treatment resulted in 77.9% survival of the plantlets. They were weaned in a glasshouse and finally moved to an agro-net shade house. 相似文献
16.
G.-Y. Kai L.-M. Dai X.-Y. Mei J.-G. Zheng W. Wang Y. Lu Z.-Y. Qian G.-Y. Zhou 《Biologia Plantarum》2008,52(3):557-560
An efficient in vitro plant regeneration system from leaves of Ophiorrhiza japonica Blume was established for the first time. Callus formation rate was more than 90.4 % from leaf segments on Murashige and
Skoog (MS) supplemented with either α-naphthaleneacetic acid (NAA) alone or in combination with 6-benzyladenine (BA). The
highest shoot regeneration (78.9 %) was achieved on MS medium containing 2.0 mg dm−3 BA and 0.2 mg dm−3 NAA, with an average of 9.4 shoots developed per leaf segment. Shoot regeneration was also improved when the leaf explants
were cultured in MS basal medium supplemented with 0.5 % (m/v) polyvinylpyrrolidone (PVP). The leaf explants from seedlings
with age of about 18–27 d showed the highest shoot regeneration. The regenerated shoots were rooted on half-strength basal
MS medium supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA), which averagely produced 24.8 roots per shoot. The plantlets were transferred to soil, where
100 % survived after 1 month of acclimatization. 相似文献
17.
Summary Plant regeneration systems from mesophyll- and cell suspension-derived protoplasts were established in Dianthus acicularis (2n=90), a species with resistance to Burkholderia caryophylli (Psedomonas caryophylli). Protoplasts were isolated from both leaves of in vitro-grown plants and cell suspension cultures established from the calluses originated from leaves of in vitro-grown plants. Protoplasts isolated from both sources showed about the same response to the type and concentration of cytokinins,
and gave the highest frequencies of cell division and colony formation in 0.1% (w/v) Gelrite?-solidified Murashige and Skoog (MS) medium supplemented with 0.5M glucose, 1.0mgl−1 (4.53 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5 mg l−1 (2.28 μM) zeatin. Numerous plantlets were regenerated after transfer of the colonies to 0.8% (w/v) agar-solidified half-strength MS
medium supplemented with 0.5 mgl−1 (2.28 μM) zeatin. Most plantlets exhibited normal phenotypes, but some showed variations, such as abnormal morphology with reduced
chromosome number, precocious flowering, and vigorous growth with a tetraploid chromosome number. Possible mechanisms responsible
for the observed somaclonal variation are discussed. 相似文献
18.
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Pistacia vera L. cv. Siirt. The in vitro procedure involved four steps that included (1) induction of shoot initials from the regenerated mature leaf tissue, (2)
regeneration and elongation of shoots from the shoot initials, (3) rooting of the shoots, and (4) acclimatization of the plantlets.
The induction of shoot initials was achieved on an agarified Murashige and Skoog (MS) medium with Gamborg vitamins supplemented
in different concentrations of benzylaminopurine (BA) and indole-3-acetic acid (IAA). The best medium for shoot induction
was a MS medium with 1 mgl−1 IAA and 2 mgl−1 BA. Numerous shoot primordia developed within 2–3 wk on the leaf margin and the midrib region, without any callus phase.
In the second step, the shoot clumps were separated from the leaf explants and transferred to a MS medium supplemented with
1 mgl−1 BA, resulting in a differentiation of the shoot initials into well-developed shoots. The elongated shoots (>3 cm long) were
rooted on a full-strength MS basal medium supplemented with 2 mgl−1 of indole-3-butyric acid in the third stage. Finally, the rooted plants were transferred to soil with an 80% success rate.
This protocol was utilized for the in vitro clonal propagation of this important recalcitrant plant species. 相似文献
19.
María Laura Vidoz Pablo Klusacek Hebe Yolanda Rey Luis Amado Mroginski 《Plant Cell, Tissue and Organ Culture》2006,86(1):111-115
In vitro protocols for plant regeneration of Arachis correntina through both somatic embryogenesis and organogenesis were developed using immature leaves as explants. Morphologically normal somatic embryos were obtained on culture media composed of 20.70 or 41.41 μM picloram (PIC) with the addition of 0.044 μM 6-benzylaminopurine (BA), resulting in a 33 and 24% of conversion into plants, respectively. The source of explants and the developmental stage of the leaves had a marked effect on somatic embryogenesis. The second folded immature leaves from in vitro growing plants were the most responsive producing up to 30% embryogenesis in MS+41.41 μM PIC. Embryos converted into plants after transfer to MS medium devoid of growth regulators and these plants were successfully acclimatised. Adventitious shoots were obtained on culture media supplemented with 2,4-dichlorophenoxyacetic acid (2,4-D) or naphthaleneacetic acid (NAA) with or without 0.044 μM BA, achieving plant regeneration in the induction media. The highest percentage of bud formation was obtained on culture medium composed of␣MS+10.74 μM NAA+0.044 μM BA (12.5%). Roots were formed on all culture media tested. Regenerated plants were transferred to pots and grew well under greenhouse conditions. 相似文献
20.
Harsh Pal Bais Julie B. Green Travis S. Walker Paul O. Okemo Jorge M. Vivanco 《In vitro cellular & developmental biology. Plant》2002,38(6):598-601
Summary
Spilanthes mauritiana DC., (Compositae), a East African medicinal herb containing pharmaceutically promising secondary metabolites, has successfully
been raised in vitro. We have developed a clonal propagation protocol that uses juvenile plants as starting material. The addition of benzylaminopurine
(BA) (1.0 μM) and naphthaleneacetic acid (NAA) (0.1 μM) to the culture medium resulted in maximum shooting response with minimal callusing. Shoots rooted best in vitro in MS medium supplemented with indole-3-acetic acid (IAA; 0.2 μM), and plants that had already developed roots showed better growth, with maximum survival rate, in the greenhouse after an
initial hardening. 相似文献