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1.
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
2.
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
3.
Marie-Claire D'Souza Madhuri Sharon 《In vitro cellular & developmental biology. Plant》2001,37(2):168-172
Summary A protocol for in vitro propagation of Bixa orellana is described. Plants were regenerated from shoot apex and nodal explants on B5 medium supplemented with 4.9 μM 2-isopentenyl adenine. The multiplication factor of shoot apex explants was higher (nine shoots per explant) than that of
the nodal explants (five shoots per explant). Regardless of the position of the nodes, all the nodal explants gave similar
responses. However, the size of the nodal explant was an important factor in producing multiple shoots: 0.5 cm nodal explants
produced the maximum multiple shoots. Regenerated shoots from shoot apex explants rooted best on MS medium supplemented with
0.05 μM α-naphthalene acetic acid (NAA). whereas shoots regenerated from nodal explants needed 2.7 μM NAA for rooting. Eighty per cent survival of in vivo transferred plants occurred on the best potting substrate, coco peat. Since the multiplication factor was nine per explant,
this protocol can be use for commercial microprogation. However, the regeneration capacity declined after 10 subcultures.
Approximately, 3350 rooted plants could be generated in 10 mo. after eight subcultures, from one shoot with a shoot apex and
four nodes. 相似文献
4.
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
5.
Summary An efficient and reproducible procedure for the large-scale propagation of Pseudoxytenanthera stocksii is described. High-frequency multiple shoot induction was achieved from nodal shoot segments collected from superior/elite
genotypes on Murashige and Skoog (MS) liquid medium supplemented with 1-naphthaleneacetic acid (NAA; 2.68 μM) and 6-benzylaminopurine (BA; 4.40 μM) at 28±1°C and 60 μmol m−2 s−1 light intensity under 12h photoperiod. In vitro-differentiated shoots were multiplied on MS liquid medium fortified with NAA (2.68 μM), BA (2.21 μM) and additives: ascorbic acid (283.93 μM), citric acid (118.10 μM), cysteine (104.04 μM), and glutamine (342.24 μM). Subculturing was carried out every 2wk on fresh shoot multiplication medium. About 125–150 shoots per culture flask were
harvested within 45–50d. In vitro-differentiated shoot clumps (three or four shoots) were successfully rooted on half-strength MS basal liquid medium with
indole-3-butyric acid (4.90 μM), BA (0.44 μM), and additives. This is the first report where in vitro- and in vivo-(through tillers) raised clonal plants were acclimatized and established in the field, where they exhibited normal growth. 相似文献
6.
Summary We have developed a highly efficient two-stage protocol for induction of multiple shoots from single node in vitro shoot tip explants of Decalepis hamiltonii. It was found that phenylacetic acid (PAA) had a synergistic effect on shoot multiplication when treated with N6-benzyladenine (BA). This protocol used PAA for both multiple shoot induction from nodal explants, elongation of primary shoots,
and initiation of adventitious shoot formation from primary shoots. Murashige and Skoog medium containing BA (2.22–31.08 μM) and α-naphthaleneacetic acid (0.27–10.74 μM) or PAA (7.34–36.71 μM) was used to initiate shoot formation from nodal explants. The maximum number of shoots per culture was produced on a medium
containing 31.08 μM BA and 14.68 μM PAA, while the longest shoot length and nodes were obtained on medium containing 22.2 μM BA and 14.68 μM PAA. Shoots subcultured on MS medium containing 22.2 μM BA and 14.68 μM PAA elongated along with secondary shoot formation. The shoots were rooted on medium containing 9.7 μM indole-3-butyric acid. The plantlets were acclimatized in soil with an 80–90% survival rate under field conditions. 相似文献
7.
Giovanni Iapichino 《In vitro cellular & developmental biology. Plant》1996,32(4):249-252
Summary Side shoots excised from underground dormant buds ofCynara scolymus L. were used as primary explants to establishin vitro cultures. A 3×3 factorial experiment with all possible combinations of three concentrations (0.5, 1.0, 2.0 mg/liter or 2.22,
4.44, 8.88 μM) ofN
6-benzyladenine (BA) and three concentrations (0, 0.1, 0.2 mg/liter or 0, 0.54, 1.07 μM) of 1-naphthaleneacetic acid (NAA) was used to determine the optimum growth regulator combination for shoot multiplication.
The highest rate of axillary shoots was induced on Murashige and Skoog agar medium supplemented with 0 mg NAA/liter and 1.0
mg BA/liter (4.44 μM). Other cytokinins tested (kinetin, zeatin, and 2-isopentenyl-adenine were less effective than BA in inducing axillary shoot
growth. Up to 60% of elongated microshoots rooted after 5 weeks on 1/2 MS agar medium supplemented with 2 mg/liter (11.42
μM) indole-3-acetic acid (IAA). Seventy percent of rooted plantlets were transferred successfully into soil. Plants are under
evaluation for their genetic uniformity and clonal fidelity. 相似文献
8.
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 for shoot regeneration (85%) and maximum number of shoots per explant
(9.5) were obtained on the 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 original 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 (IBA) after 25 d of culture. Fifty percent of shoots were also directly rooted as microtuttings on
a peat moss, soil, and compost mixture (1∶1∶1). About 52% of plantlets were successfully acclimatized and established in pots. 相似文献
9.
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
10.
Summary Shoots by direct and indirect organogenesis and somatic embryos were induced from tubercles excised from Ariocarpus kotschoubeyanus seeds germinated in vitro. Shoot formation was greatest (6.3 per explant) when explants were cultured on Murashige and Skoog (MS) medium supplemented
with 6-benzylaminopurine (BA; 13.3 μM) and α-naphthaleneacetic acid (NAA; 5.4 μM). Individualized shoots were rooted in half-strength MS; the addition of activated charcoal (1 g l−1) and the use of sun cap closures to seal containers improved the rooting of shoots. Nearly 20% of the explants produced somatic
embryos on media containing combinations of BA (8.9–22.2 μM) and NAA (0.5–5.4 μM). The establishment of plantlets in soil presented no significant problems. In vitro culture is a useful option for mass propagation of A. kotschoubeyanus and contributes to its conservation. 相似文献
11.
C. S. Lin M. C. Tseng P. I. Hong W. C. Chang 《In vitro cellular & developmental biology. Plant》2006,42(4):331-335
Summary Inflorescence proliferation is a plant tissue culture technique that, can be used to obtain in vitro inflorescences year-round without the intervening development of vegetative organs. In this study, we used albino mutant
inflorescences of Dendrocalamus latiflorus as the original explant material to investigate, the effect of plant growth regulators on long-term inflorescence proliferation.
The albino inflorescences proliferated on solidified Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ),
and the optimal concentration for successful long-term inflorescence proliferation was 0.45 μM TDZ. A combination of α-naphthaleneacetic acid (NAA) with 0.45 μM TDZ inhibited the inflorescence proliferation. Inflorescences cultured on a TDZ-free medium supplemented with 26.82 μM NAA rooted in 21 d, vegetative shoots formed by 42 d and, in one case, flowering occurred after 63 d. The auxins 2,4-dichlorophenoxyacetic
acid (2,4-D, 4.52 μM) and pieloram (4.14 μM) induced shoot formation. The protocol described can be used to produce large numbers of mutant inflorescences within a relatively
short period of time. 相似文献
12.
Rakhi Chaturvedi S. P. Bhatnagar 《In vitro cellular & developmental biology. Plant》2001,37(2):255-258
Summary A protocol for in vitro shoot regeneration from cotyledon explants of Citrullus lanatus (Thunb.) Matsum. & Nakai cv. Sugar Baby is described. The cotyledons excised from 7-d-old aseptic seedlings showed the highest
percentage of shoots on Murashige and Skoog (MS) + N6-benzyladenine (BA; 3.0 μM) + N6-[2-isopentenyl] adenine (2iP; 3.0 μM) and MS + BA (3.0 μM) + indole-3-acetic acid (IAA; 3.0 μM). Whereas the latter medium induced shoot regeneration after the callusing of the explant, the former stimulated direct shoot
formation. The regenerated shoots were rooted and the resulting plants were established in earthen pots with 55% success. 相似文献
13.
Summary A procedure for micropropagation of pitaya using thidiazuron (TDZ) and naphthaleneacetic acid (NAA) is described. Explants
were excised from young joints of mature plants and cultured on Murashige and Skoog medium (MS) containing 0.5 μM NAA and 0.5 μM TDZ. Shoots produced from these first explants were cut up to produce secondary explants, either by decapitation or by longitudinal
division into three parts. The decapitated and longitudinal explants were cultured on MS supplemented with 0.5 μM NAA and either 0.01, 0.09, 0.5, or 0.9 μM TDZ. Decapitated explants produced more shoots at higher frequency that the longitudinal explants. For both types of secondary
explants, most shoots were developed from the distal parts. Shoots produced from secondary explants were rooted in MS and
then transferred to soil where they produced normal plants. 相似文献
14.
Summary Success has been achieved in developing a complete protocol for mass propagation of Anogeissus pendula and A. latifolia, two important forest species found in India. Seeds cultured on plant growth regulator-free, semisolid Murashige and Skoog
(MS) medium germinated within 5–6 wk and formed 4–6-cm long shoots. The shoots multiplied on MS+4.4 μM benzyladenine (BA)+5.7 μM indoleacetic acid (IAA) + casein hydrolysate (100 mgl−1) + ascorbic acid (50 mgl−1) + sucrose (3%) + agar (0.8%). A majority of the genotypes rooted with more than 90% efficiency when 5–6 cm individual shoots
were cultured on 1/2MS (only major salts reduced to half strength)+2.3 μM IAA+2.5 μM indolebutyric acid (IBA) + sucrose (3%)+agar (0.8%) for 15 d. Those 10% (approx.) genotypes that did not root well on the
above medium could be rooted with ease by increasing the concentration of IAA in the rooting media from 2.3 to 5.7 μM. The in vitro-raised plants were successfully transferred to the soil with a success rate of over 85%. Using this protocol, over 560 000
tissue-cultured plants of these two species have been produced and dispatched to various state forest departments for field
trials and routine plantations. 相似文献
15.
S. Pattnaik C. Pradhan S. K. Naik P. K. Chand 《In vitro cellular & developmental biology. Plant》2000,36(5):407-411
Summary A complete and efficient protocol is presented for plant regeneration from cell-suspension cultures of Dalbergia sissoo Roxb., an economically important leguminous tree. Factors influencing callus initiation, establishment of cell-suspension
culture, callus formation from embredded microcolonies, and shoot organogenesis from suspension-derived callus were identified.
Of the two different auxins tested, callus induction was better on a medium containing naphthalene acetic acid (NAA). The
percentage of callus induction increased considerably when NAA at 2.0 mg l−1 (10.8 μM) was added in conjunction with 0.5 mg l−1 (2.2 μM) N6-benzyladenine (BA). Of the three different explants evaluated for callus induction, hypocotyl segments were most responsive.
Friable hypocotyl-derived callus from the second subculture passage was used to initiate the cell-suspension culture. Optimum
growth of the cell suspension was observed in MS medium supplemented with the same growth regulators as described above for
callus induction, with an initial inoculum cell density of 1%. The plating efficiency of the microcolonies was greatly influenced
by harvesting time and the gelling agent used for plating. Efficiency was highest (93%) with cells harvested at their exponential
growth phase and plated in 1.2 g l−1 Phytagel. Shoot organogenesis from callus cultures was higher on a medium supplemented with a combination of BA and NAA than
on BA alone. Seventy-one per cent of cultures exhibited shoot-bud differentiation on a medium containing 3.0 mg l−1 (13.3 μM) BA and 0.5 mg l−1 (2.7 μM) NAA. Regenerated shoots were rooted on half-strength MS medium containing 1 mg l−1 each of indole-3-acetic acid (5.7 μM), indole-3-butyric acid (4.9 μM) and indole-3-propionic acid (5.3 μM). Plantlets were acclimated and established in soil. 相似文献
16.
Marián López José Pacheco Roberto Rodríguez Ricardo J. Ordás 《In vitro cellular & developmental biology. Plant》1996,32(2):109-114
Summary Plantlet regeneration of salgare?o pine (Pinus nigra Arn. ssp.salzmannii (Dunal) Franco) was achieved from cotyledons. The data showed that the best differentiation response occurred when cotyledons,
from 1-d-old embryos germinated in darkness, were cultured on Murashige and Skoog medium (half-strength macroelements) with
22 μM N6-benzyladenine (BA) and 0.05 μM α-naphthaleneacetic acid (NAA) for 2–3 wk. Shoot development was obtained by subculturing treated explants on the same medium
without growth regulators. Shoots were successfully micropropagated by sequential subculturing them on medium containing growth
regulators (5 μM BA and 0.005 μM NAA) and on hormone-free medium for 2 and 12 wk, respectively. To induce adventitious roots, shoots were treated with 3 μM NAA, and 8 μM indole-3-butyric acid for 2 wk, followed by transfer to Murashige and Skoog medium (1/4-strength macroelements, 20 g·L−1 sucrose) without growth regulators. After 3–4 wk, almost all the rooted shoots (65%) could be successfully transplanted and
acclimatizated in the greenhouse, where the plants exhibited normal growth habit. 相似文献
17.
L. Burdyn C. Luna J. Tarracó P. Sansberro N. Dudit A. Conzález L. Mrocinski 《In vitro cellular & developmental biology. Plant》2006,42(3):235-239
Summary Adventitious bud regeneration from leaf and internode explants of Aloysia polystachya was achieved. Shoots from nodal segments grown in vitro were cut into pieces and used as sources of explants. Organogenesis was induced from both explants cultured on quarter-strength
Murashige and Skoog (MS) semisolid medium (plus sucrose 5 g l−1) containing different combinations of 6-benzyladenine (BA) and α-naphthaleneacetic acid (NAA) under 116 μmol m−2 s−1 photosynthetic photon flux density (PPFD), 14-h photoperiod, and at a temperature of 27±2°C. The type of explant markedly
influenced organogenesis and growth of the regenerated shoots. The regeneration frequencies were higher with leaf explants,
while the number of shoots formed per responsive explant was greater with internode explants. However, the growth of regenerated
shoots from internodes was seriously affected by vitrification. The number of shoots produced per responsive leaf explant
increased from one to seven as the percentage of leaf explants producing shoots increased from 20 to more than 80%. NAA at
0.05 μM in combination with BA at 0.5μM induced the highest regeneration rate (87±8.8%) after 20 d of culture, yielding 5.9±0.8 shoots per responsive leaf explant.
Histological examination confirmed the occurrence of direct organogenesis. The regenerated shoots from the best induction
treatment were transferred to a fresh medium without plant growth regulators for 30 d. Finally, the elongated shoots were
rooted by pre-treatment in an aqueous solution of NAA at 500 μM for 2 h and transferred to 1/4 MS. All plantlets raised in vitro were phenotypically normal and successfully hardened to ex vitro conditions. An experimental field plot with 2-yr-old in vitro-regenerated plants was established. 相似文献
18.
Summary We have developed efficient methods for plant regeneration, via both embryogenesis and organogenesis, of Smooth Cayenne pineapple,
Ananas comosus (L.) Merr. Leaf bases and core (stem) sections of in vitro shoots, produced from culture of crown tip meristem, were used as explants for plant regeneration as follows: (1) Leaf base
and core section explants cultured on Murashige and Skoog (MS) medium containing 41 μM 4-amino-3,5,6-trichloropicolinic acid (picloram, P) or thidiazuron (T)/P combinations produced embryogenic tissues. Different
types of embryogenic tissues (friable emryogenic tissue, embryogenic cell cluster, and chunky embryogenic tissue) have been
developed with varying properties in terms of growth rate and state of development. The embryogenic tissues regenerated shoots
upon culture on MS medium containing 13 μM 6-benzylaminopurine (BA) and 1μM α-naphthaleneacetic acid (NAA) followed by culture on MS medium containing 4 μM BA. (2) Crown tip meristems cultured on MS medium containing 13 μM BA followed by leaf explants cultured on MS medium with 27 μM NAA and 1 μM BA produced shoots via direct organogenesis. (3) Explants cultured on MS medium containing 5 μM T and 0.5 μM indole-3-butyric acid (IBA) produced nodular globular structures, which produced shoots upon culture on MS medium containing
1 μM BA and 1 μM gibberellic acid. Shoots obtained from all of the above methods were rooted in half-strength MS medium containing 3 μM NAA and 2.5 μM IBA. Plants were transferred to the greenhouse or shipped to Costa Rica for field trials. Somatic embryo-derived plants exhibited
21 % spininess, and organogenic-derived plants exhibited 5% spininess in the field trials. 相似文献
19.
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
20.
Summary Shoot tips and leaves excised from in vitro shoot cultures of Salvia nemorosa were evaluated for their organogenic capacity under in vitro conditions. The best shoot proliferation from shoot tips was obtained on Murashige and Skoog (MS) medium supplemented with
8.9 μM 6-benzylaminopurine (BA) and 2.9 μM indole-3-acetic acid (IAA). Leaf lamina and petiole explants formed shoots through organogenesis via callus stage and/or
directly from explant tissue. The highest values for shoot regeneration were obtained with 0.9 μM BA and 2.9 μM IAA for lamina explants. No shoot organogenesis was obtained on leaf explants cultured on MS medium supplemented with α-naphthaleneacetic
acid (NAA). The regenerated shoots rooted the best on MS medium containing 0.6 μM IAA or 0.5 μM NAA. In vitro-propagated plants were transferred to soil with a survival rate of 85% after 3 mo. 相似文献