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1.
In vitro propagation technique of Balanites aegyptiaca, a multipurpose woody tree was studied. Nodal segments including axillary bud from mature tree were used as an explant and
their morphogenetic potential was tested on MS media with various concentrations (2.5–15.0 μM) of 6-benzyladenine (BA), Kinetin,
and Thidiazuron alone or in combination with different concentrations (0.5–2.5 μM) of α-naphthalene acetic acid (NAA). Nodal
segments showed axillary bud proliferation in almost all media tried. MS medium containing 12.5 μM BA alone was effective
for inducing multiple shoots (5.0 ± 0.22) with an average shoot length (3.7 ± 0.26 cm) in 67% of cultures. A better shoot
differentiation and elongation was achieved in a combined treatment of BA (12.5 μM) and NAA (1.0 μM). Half strength MS medium
supplemented with Indole-3-butyric acid (IBA) gave the best result for rooting. The maximum frequency of root formation (68%),
number of roots (5.3 ± 0.32) and root length (4.1 ± 0.38 cm) was obtained on half strength MS medium containing 1.0 μM IBA.
The regenerated plantlets were potted and acclimatized successfully in a growth chamber and then moved to the greenhouse. 相似文献
2.
This report describes in vitro shoot induction and plant regeneration from nodal segments of Balanites aegyptiaca on Murashige and Skoog (MS) medium fortified with 6-benzyladenine (BA), thidiazuron (TDZ) and kinetin (Kin) (0.5–20.0 μM).
MS medium supplemented with BA (12.5 μM) was the most effective in inducing bud break and growth and also in initiating multiple
shoot proliferation. However, the optimal level of TDZ supplementation to the culture medium was 5.0 μM. Shoot cultures were
established by repeatedly subculturing the original nodal explants on the same medium. Highest number of shoots (11.5 ± 0.7)
and shoot length (5.0 ± 0.2 cm) were achieved when cultures were subcultured on MS medium supplemented with 12.5 μM BA and
1.0 μM α-naphthalene acetic acid (NAA). The shoots regenerated from TDZ supplemented medium when subcultured to hormone free
MS basal medium considerably increased the rate of shoot multiplication and shoot length by the end of fifth subculture. Rooting
of the shoots was achieved on MS medium augmented with 1.0 μM indole-3-butyric acid (IBA) plus 0.5% activated charcoal followed
by their transfer to half strength MS basal medium. The in vitro raised plantlets with well developed shoots and roots were
successfully established in earthen pots containing garden soil and were grown in greenhouse with 70% survival rate. The results
of this study provide the first successful report on in vitro direct plant regeneration of B. aegyptiaca. 相似文献
3.
Different nutrient media can affect in vitro culturing protocols, and experimentation under varied growth conditions is valuable in plants where in vitro methods are in preliminary stages. We carried out the first in vitro propagation studies for the endangered species Caragana fruticosa (Fabaceae). We evaluated various nutrient media for their impact on shoot elongation and axillary bud proliferation using
different concentrations of 6-benzylaminopurine (BA) and α-naphthaleneacetic acid (NAA). Shoot elongation was evaluated based
on adventitious shoot primary culture and subculture regeneration from Caragana seedlings. Our goal was to improve both micropropagation and regeneration in C. fruticosa. MS nutrient media was superior to 1/2MS macronutrients, DKW, QL, and WPM for shoot elongation and axillary shoot proliferation.
Shoots grown on 1/2MS and WPM exhibited some chlorosis, and shoots on QL produced larger leavers than plants growing on normal
medium. The shoot proliferation coefficient on MS media supplemented with 2.22 μM BA and 0.44 μM BA + 2.69 μM NAA was significantly
higher than that with other treatments in the primary culture. Shoots on 2.22 μM BA showed a higher proliferation coefficient
(3.17) than others in the subculture. Shoots were rooted on 1/2MS medium with the addition of different concentrations of
NAA. The optimal concentration for rooting was 0.27 μM NAA (74%). Roots exhibited many stout and long root hairs. Survivl
of established plantlets was 82% at 30 days after transfer to soil. Plants established in the green house showed normal growth
and displayed no apparent morphological differences compared to stock plants. 相似文献
4.
A micropropagation method for Jaal (Salvadora persica)—a tree of arid horticulture and forestry has been developed. Nodal segments of fresh shoot sprouts originated from axillary
buds obtained from a plant around 35–40 years old lopped plant were used as explants for establishment of in vitro cultures.
Surface-sterilized explants produced optimum number of shoots through activation of axillary buds on Murashige and Skoog’s
(MS) medium containing 8.88 μM BA (6-benzyladenine) + additives (25 mgl−1 each of adenine sulphate, arginine, citric acid, 50 mgl−1 ascorbic acid). The shoot multiplication was influenced by the successive transfer of the mother explants for 4–5 passages.
The maximum number (23.1 ± 0.73 shoots per explant) of shoots were regenerated on MS supplemented with 1.11 μM BA + 1.16 μM
Kn (Kinetin) + 0.54 μM NAA (α-naphthalene acetic acid). About 90% shoots pulse-treated with a combination of 2460.27 μM Indole-3-butyric
acid (IBA) + 494.56 μM NOA (2-naphthoxy acetic acid) were rooted ex vitro on soilrite within 15–18 days. Over 80% cloned plantlets
were hardened successfully in a green house and transferred to polybag/pots. 相似文献
5.
A method for efficient in vitro regeneration of Leucaena leucocephala cv. K636 was developed using immature zygotic embryos as the explant. Multiple shoot induction was achieved by culturing
the explants on half-strength Murashige and Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.03 to 1.5 μM). The maximum
number of shoots per explants was achieved in 0.26 μM TDZ-supplemented media. TDZ concentration significantly influenced the
induction of multiple shoots as well as the shoot-length. TDZ at 0.35 μM or higher concentrations resulted in abnormal stunted
shoots. Full-strength MS media devoid of any plant growth regulator were used for shoot elongation. In vitro root induction
was achieved in half-strength MS media supplemented either with 0.54 μM 1-naphthaleneacetic acid (NAA) or with 14.76 μM indole-3-butyric
acid (IBA) in combination with 0.23 μM kinetin (Kn). Media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn induced
twofold–threefold more roots than media supplemented with 0.54 μM NAA. However, the average root length was lower in 14.76 μM
IBA in combination with 0.23 μM Kn than in 0.54 μM NAA. Regenerated plants were maintained under normal condition after hardening.
Plants, which were rooted on media supplemented with 14.76 μM IBA in combination with 0.23 μM Kn showed higher survival rate
during hardening than those rooted on 0.54 μM NAA supplemented media. 相似文献
6.
In-Sun Park Momoko Koiso Satomi Morimoto Takafumi Kubo Hyun-O Jin Ryo Funada 《Journal of Wood Science》2012,58(1):64-68
We attempted to develop a method for the regeneration of plantlets from mature seeds of medically important Magnolia obovata via the induction of somatic embryogenesis in vitro. We initially cultured halves of mature seeds on either Murashige and
Skoog (MS) medium or B5 medium that contained 0, 1, 5 or 10 μM gibberellic acid (GA3) for 1 month and then transferred the half-seeds to half-strength MS basal medium or B5 basal medium for further culture in the absence of GA3. Proembryogenic masses (PEMs) were observed 1 month after the transfer of the halved mature seeds to the medium without GA3. The frequency of formation of PEMs was higher (28%) after initial culture in MS basal medium plus 1 μM GA3 than in other tested media (0 or 4%). Somatic embryos that had been developed from PEMs were cultured on half-strength MS
basal medium or B5 basal medium for completion of maturation and then transferred to fresh aliquots of the same medium for initiation of germination.
The frequency of germination, with the formation of normal primary leaves and roots, was above 80%. We transferred the somatic
embryo-derived plantlets to soil for acclimatization and the plantlets continued to thrive. 相似文献
7.
An efficient regeneration protocol for rapid mass propagation and uptake of heavy metals in Albizia lebbeck (L.), a fast growing, medicinally as well as economically important timber yielding tree was developed. Nodal segments derived
from a 20-year-old tree were cultured on MS (Murashige and Skoog) medium supplemented with 10 μM 6-Benzyladenine (BA) and
1 μM α-Naphthalene acetic acid (NAA) showed optimum shoot regeneration frequency (76.6%), number of shoots (23.2 ± 0.28) per
explant and shoot length (2.86 ± 0.08 cm) after 10 weeks of culture. After standardizing a reliable protocol for micropropagation,
effects of ZnSO4 (0.06–0.48 mM), CuSO4 (0.02–0.2 mM) and CdCl2 (0.0001–0.001 mM) on shoot morphogenesis were also assessed. The regenerated shoots maintained on maintenance medium (MS + 10.0 μM
BA + 1.0 μM NAA) containing ZnSO4 (0.06 mM) showed maximum response in terms of shoot number (24.5 ± 0.83) and length (5.9 ± 0.05 cm) after 10 weeks of culture.
Proline content showed an increasing trend while chlorophyll (a and b) content exhibited decreasing trend with an increased
metal concentrations compared to MM cultures, and maximum increase in proline and decrease in chlorophyll content was recorded
in cultures grown on Cd-enriched medium. Best rooting was accomplished on half strength MS medium with 2.0 μM IBA and ZnSO4 (0.06 mM). The plantlets thus obtained were successfully hardened and transferred to greenhouse with 75% survival rate and
exhibited normal morphological characteristics compared to donor plant. 相似文献
8.
An efficient and improved shoot regeneration technique for the micropropagation of Vitex negundo, an aromatic and medicinal shrub through in vitro culture of nodal segments with axillary buds, is described. Thidiazuron
(TDZ) used at 1.0 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation
at the rate of 25 microshoots per nodal explant with axillary buds, after 4 weeks of culture. By repeated subculturing of
nodal explants, a high-frequency multiplication rate was established. Optimum shoot multiplication and elongation was achieved
when TDZ exposed explants were subcultured on Murashige and Skoog (MS) media containing a combination of 1.0 μM 6-benzyladenine
(BA) and 0.5 μM α-naphthalene acetic acid (NAA). Efficient rooting was achieved directly in soilrite when basal portion of
the shoots were treated with 500 μM indole-3-butyric acid for 10 min which was the most effective in inducing roots, as 97%
of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to
ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered. No phenotypical differences for morphogenesis
were observed among the regenerants. 相似文献
9.
Hamako Sasamoto Yohichi Wakita Shinso Yokota Nobuo Yoshizawa 《Journal of Forest Research》2000,5(4):265-270
An improved method for selection of large electro-fused protoplasts ofPopulus alba by a micromanipulator was developed. The conditions for electric cell fusion treatment were optimized. For the best result,
protoplasts with a cell density of 5 × 105/mL were treated with an alternate current (1 MHz, 200 V/cm) and pulsed with a direct current (2 kV/cm) for 100μs in 2.5 mM CaCl2 and 0.55 M mannitol. The electo-fused protoplasts were cultured in NH4NO3-free Murashige and Skoog’s medium containing 0.6 M of mannitol, 0.09 M sucrose, 1μM of 2,4-dichlorophenoxyacetic acid and 0.1μM of benzyladenine, the same medium used for protoplast culture, but at a very low cell density of 5–10 × 102/mL in a well of a 96-well culture plate. When cell aggregates derived from individual fused protoplasts were transferred
to fresh medium with 0, 0.3 or 0.6 M mannitol, large colonies developed. In the shoot differentiation medium, the reaction
of the calluses derived from large fused protoplasts towards the growth regulators differed from the non-fused ones. In medium
containing 1μM each of naphthalene acetic acid andN-(2-chloro-4-pyridyl)-N′-phenylurea, growth of callus from electro-fused ones was not reduced by much compared to the control, but shoot differentiation
was inhibited. Gibberellic acid (0.1–10μM) was beneficial to shoot regeneration; however, irregularly shaped leaves appeared at high gibberellic acid concentrations.
Shoots regenerated were rooted in Murashige and Skoog’s medium containing 4μM of indole-3-butyric acid. Some plantlets obtained had a varied morphology. Based on the characteristics of growth, some
cells derived from electro-fused protoplasts appear to be physiologically different from the non-fused one. 相似文献
10.
Qingbin Jiang Yong Zhang Chonglu Zhong Bingshan Zeng Didier Bogusz Claudine Franche 《New Forests》2012,43(2):143-154
An in vitro plant regeneration protocol via indirect organogenesis from morphogenetic callus was established for Casuarina cunninghamiana Miq. Effects of plant growth regulator NAA (naphthaleneacetic acid) and BAP (6-benzylaminopurine), sucrose and AgNO3 on callus induction, adventitious bud differentiation and shoot development were examined. Explants used were epicotyl fragments
from 45-day-old seedlings. The largest callus (4.29 mm in diameter) was obtained after 1 month on a basic culture medium consisting
of Murashige and Skoog ? macro- and full strength micro- elements, Nitsch and Nitsch vitamins, supplemented with 0.54 μM NAA,
3.30 μM BAP, and 30 g L−1 sucrose. The calli were subcultured in the same medium above for 2 months. They were then cultured for another 2 months for
adventitious bud differentiation and shoot development. The highest mean adventitious bud differentiation, number of shoots
formed per callus and number of shoots ≥2 cm long per callus (47.50%, 27.38 and 4.75, respectively) were achieved on the above
medium modified with NAA at 0.27 μM and supplemented with AgNO3 1 mg L−1. Shoots were successfully rooted without plant growth regulator and the rooted plantlets survived and grew normally. This
protocol for in vitro plant regeneration provides a tool not only for vegetative propagation but also for plant genetic transformation
and gene function studies of C. cunninghamiana. 相似文献
11.
A highly reproducible and efficient in vitro shoot regeneration system was developed in a potential medicinal plant, Albizia lebbeck using root explants. Root explants from 15 day-old-aseptic seedlings were cultured on Murashige and Skoog (MS) medium supplemented
with different concentrations (0.5, 2.5, 5.0, 7.5 and 10.0 μM) of 6-Benzyladenine (BA), Kinetin (Kn), 2-Isopentenyl adenine
(2-iP) singly as well as in combination with α-Naphthalene acetic acid (NAA) (0.1, 0.5, 1.0, 1.5 and 2.0 μM). The highest
rate of shoot multiplication (16.0 ± 1.87 for the average shoot number and 5.16 ± 0.38 cm for shoot length) was achieved on
MS medium supplemented with 7.5 μM BA and 0.5 μM NAA. The effects of medium type, medium strength, pH and subculture on shoot
induction and proliferation were also tested. An average of 21.6±2.87 shoots per explants could be obtained following this
protocol. Rooting was achieved on microshoots using half strength MS medium with 2.0 μM Indole-3-butyric acid (IBA) after
four weeks of culture. The in vitro raised healthy plantlets were successfully established in earthen pots containing garden soil and grown in greenhouse with
>80% survival rate. 相似文献
12.
Crataeva magna (Lour.) DC (synonym C. nurvala) is a high-value Indian medicinal tree. The multiple uses of C. magna have resulted in its over-exploitation. Erratic seed germination combined with destructive harvesting and habitat destruction
in the form of deforestation has added to the enormity of the problem. Therefore, the need for conservation of this plant
is vital. Development of an efficient micropropagation protocol will play a significant role in meeting the requirement of
quality planting material for commercial cultivation thereby conserving the species in its natural habitat. In the present
study, shoot multiplication was achieved by culturing single node segments derived from a field grown tree on Murashige and
Skoog’s (MS) medium supplemented with 2.66 μM N6-benzyladenine, 1.39 μM Kinetin (Kn), 0.57 μM indole-3-acetic acid (IAA),
3% sucrose and 0.2% gelrite. 96% rooting was achieved within 22 days by culturing the in vitro formed shoots on half strength
MS medium with 11.42 μM IAA, 9.8 μM indole-3-butyric acid, 0.46 μM Kn and 198.25 μM phloroglucinol. Following a simple hardening
procedure involving sequential transfer of plants to a greenhouse, polyhouse, and shade net, the tissue-cultured plants were
transferred to the field where the survival rate was 100%. To this date 500 plants have been produced. Inter simple sequence
repeat analysis has confirmed genetic uniformity of the tissue-cultured plants. 相似文献
13.
Kavita Arora Meena Sharma Jyoti Srivastava S. A. Ranade A. K. Sharma 《Agroforestry Systems》2010,78(1):53-63
An efficient in vitro process for rapid clonal propagation of a 40-year-old tree of Azadirachta indica employing nodal stem segments was developed. Season of collection and maturity of explants showed direct influence on bud-break.
Nodal stem segments collected during the month of April gave best response. Maximum bud-break (78.6–81%) was obtained when
middle order nodes (3rd or 4th node from apex) were taken. Amongst different cytokinins used, 6-benzylaminopurine (BAP) at
the concentration of 1.11 μM was found most effective in inducing multiple shoots, whereas inorganic and organic constituents
of the medium influenced growth and general condition of proliferating shoots. On an average 3.1 shoots per explant were regenerated
in modified Murashige and Skoog medium supplemented with 1.11 μM BAP, 1.43 μM indole-3-acetic acid (IAA) and 81.43 μM adenine
hemisulphate. Isolated shoots were rooted in presence of 2.46 μM indole-3-butryic acid (IBA). Root induction took place in
8–10 days with 100% rooting. The in vitro-raised plantlets were successfully transplanted in potted soil and finally grown
under field conditions with 100% survival. The genetic fidelity of such in vitro-raised field-grown plants was ascertained
by random amplified polymorphic DNA (RAPD) markers. Furthermore, the azadirachtin content of in vitro-cloned plants was found
comparable to the mother tree proving their chemical stability also. The protocol developed holds good for in vitro cloning
of mature elite neem trees. 相似文献
14.
Thidiazuron (TDZ) induced somatic embryogenesis from immature zygotic embryos in Cinnamomum pauciflorum Nees while 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA) or picloram only induced callus and/or adventitious
buds. The highest induction frequency for somatic embryogenesis was achieved with MS medium (Murashige and Skoog in Physiol
Plant 15:473–497 1962) supplemented with 2.5 μM TDZ using torpedo-shaped embryos (3–5 mm in length) as explants. In addition, induction medium
was supplemented with 0.8 g l−1 casein, 0.4 g l−1 glutamine, and 10 g l−1 sucrose. Somatic embryos (SEs) initiated from root tips or hypocotyls without callus formation. SEs were maintained and multiplied
via secondary somatic embryogenesis. Embryo maintenance medium was similar to induction medium except that TDZ was reduced
to 0.5 μM. Secondary embryogenesis was enhanced by supplementation of 5 g l−1 activated charcoal in the culture. The best medium for embryo maturation was MS medium containing 30 g l−1 sucrose and 5 g l−1 Phytagel without plant growth regulators. A typical mature SE consisted of two large cotyledons and a short embryo proper.
Approximately 82% of selected mature SEs were able to germinate and 63% could convert into plantlets on germination medium
that was composed of half strength MS medium salts, 10 g l−1 sucrose, 3 g l−1 Phytagel, and 5 g l−1 activated charcoal. 相似文献
15.
An in vitro clonal propagation procedure for mature Tectona grandis (teak) trees is described. Multiple shoots were induced from nodal segments through axillary bud proliferation. A shoot multiplication
rate of 6.33 was achieved on Murashige and Skoog's (MS) medium supplemented with 10 μM 6-benzyladenine BA and 1 μM 1-naphthalene
acetic acid (NAA) during every subculture cycle of 4 weeks. In vitro raised shoots could be successfully rooted (66.66%) on
liquid MS medium supplemented with 15 μM NAA, with 1.60 roots per shoot, every 6 weeks of culture. In vitro hardening was
carried out in sand soaked with half-strength MS medium (organic free). The plantlets were acclimatized first in a mist chamber
and then in polybags in a mixture of soil, sand, and farmyard manure (1 : 1 : 1 v/v) in a shade house. 相似文献
16.
Using Agrobacterium-mediated gene transfer, we generated transgenic hybrid sweetgum (Liquidambar styraciflua × L. formosana) overexpressing two types of genes to enhance plant remediation of mercury-contaminated soil and water: bacterial γ-glutamylcysteine
synthetase gene (ECS), the first and most important enzyme in phytochelatin synthesis, or various genes encoding a mercuric ion reductase (merA9, merA18, merA77). Hybrid sweetgum proembryogenic masses (PEMs) constitutively overexpressing ECS were able to grow in the presence of 50 μM HgCl2, which inhibited wild-type PEMs, but plantlets regenerated from the PEMs had abnormal form and did not survive for more than
a few weeks following germination. In contrast, mature somatic embryos generated from PEMs constitutively overexpressing merA9 and merA18 converted to normal plantlets on germination medium containing 25 μM HgCl2, while control embryos were killed on 25 μM Hg(II)-medium. Transgenic merA plantlets displayed enhanced resistance to Hg(II) and released Hg(0) two to three times more efficiently than the wild-type
plantlets. 相似文献
17.
Ekambaranellore Prakash Patan Shaik Sha Valli Khan Thoguru Jonh Vivek Sreenivasa Rao Elu Singh Meru 《Journal of Forest Research》2006,11(5):329-335
In vitro clonal multiplication of Pterocarpus santalinus L. was achieved using mature nodal explants of a 10-year-old elite quality tree. Combinations of serial transfer technique
and incorporation of antioxidants (250 mg/l L-ascorbic acid and 50 mg/l citric acid) into the culture medium helped to minimize medium browning and improve explant survival
during shoot sprouting. About 70% of explants were sprouted on Murashige and Skoog (MS) liquid medium containing 4.4 μM 6-benzyladenine (BA). The explant harvest period also influenced the bud break and shoot sprouting in nodal explants. The
combination of 4.4 μM BA and 2.2 μM thidiazuron (TDZ) was found to be the most suitable growth regulator for obtaining the highest percentage of nodal segment
sprouting (74%–75%), the number of secondary shoots per primary shoot (two or three), the shoot length (5–6 cm), the number
of new nodal segments generated per active explant (four or five), and the multiplication coefficient (3.5) within 6 weeks.
Repeated subculturing of nodal explants obtained from shoot cultures enabled continuous production of healthy axillary shoots.
At the end of the sixth passage, about 90% of nodal explants produced five or six healthy green shoots, each being about 6.6 cm
long with six or seven nodes. Multiplication coefficient was also increased from the first subculture (5.4) to the sixth subculture
(8.3). The best rooting response was achieved on solidified half-strength MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA). About 70% of the micropropagated plantlets were established successfully in 20-cm pots containing
a mixture of soil and farmyard manure (4 : 1 ratio) and formed new leaflets. 相似文献
18.
Cottonhead windhairdaisy (Saussurea laniceps Hand.-Mazz.) is one of the most famous and important medicinal herbs in China. Illegal collection from wild populations is increasingly threatening the present environment of S. laniceps. Establishment of an efficient method for micropropagation is the best way to change its endangered situation. When mature seeds of S. laniceps were cultured on hormone-free MS medium, plantlets were formed from germinated seeds in 7–10 d. Then 0.5 cm × 0.5 cm leaf explants were transplanted to MS medium supplemented with 1-naphthalene-acetic acid (NAA)/2,4-D and benzyladenine (BA)/KT and callus was achieved 10 d after transfer. Shoot bud regeneration occurred from callus cultured on MS medium supplemented with different growth regulators 20 d after culturing. The regeneration percentages varied with the different components of plant growth regulators. The percent regeneration from callus pretreated at low temperature of 5°C increased significantly compared with those incubated at 23/20°C directly. Optimal regeneration was observed with explants on media supplemented with 1.5 mg8226;L–1 BA plus 0.2 mg8226;L-1 NAA. In the presence of 0.2 mg8226;L–1 NAA in half-strength MS, 78% of the shoots formed roots. Plantlets from explants showed 63% survival after acclimatization. 相似文献
19.
A rapid and efficient method for the regeneration of plantlets from root explants ofRobinia pseudoacacia L. by suspension culture was established. The roots taken from aseptically grown 15-day-old seedlings were used as explants.
It was determined that photoperiodicity was necessary for root proliferation, and that the promotive effect of IAA (3-indoleacetic
acid) on root proliferation was better than that of IBA (3-indolebutyric acid). The roots cultured in 1/2 MS liquid medium
containing 3 μM IAA and 1% sucrose at 25°C under 16-hour photoperiod with 50 μmol m−2s−1 PPFD (photosynthetic photon flux density) shaking at 100 times/min reciprocally showed high efficiency for root proliferation.
BAP (6-benzylaminopurine) was found to be essential to induce adventitious shoots from the roots, and the roots cultured in
the medium supplied with 3 μM BAP combined with 1–6 μM IAA for 3 weeks under the same conditions as in the root proliferation
period were most suitable for adventitious shoot inducement. 相似文献
20.
T. D. Salla C. dos S. Silva K. L. de G. Machado L. V. Astarita E. R. Santarém 《林业研究》2018,29(3):623-629
Eucalyptus is very recalcitrant to in vitro culture. In this research, an efficient shoot organogenesis system was developed using 60-day-old plants of Eucalyptus globulus grown in vitro and non-aerated liquid medium to improve shoot proliferation. Cultures were initiated with hypocotyls and leaf segments from plantlets cultivated on semisolid ½ MS modified medium supplemented with 4.44 µM 6-Benzyladenine (BA) and 16.1 µM 1-Naphthaleneacetic acid (NAA). Calli were transferred to shoot induction medium, with either 0.5 or 2.7 µM NAA. Shoot multiplication was carried out on 4.44 µM BA + 0.5 µM NAA medium, and semisolid and non-aerated liquid systems were compared for improving shoot proliferation. Rooting of adventitious shoots was evaluated on medium containing NAA or Indole-3-butyric acid -IBA (5 and 16 µM). Callogenesis was obtained from both types of explants, although shoot formation was only obtained from leaf-derived calli. Shoot proliferation on 4.44 µM BA + 0.5 µM NAA resulted in the most shoots/callus. Non-aerated liquid medium was more efficient in promoting shoot multiplication (53.5 shoots/callus) than was semisolid medium (28.5 shoots/callus). Levels of phenolic compounds were significantly reduced in the shoots cultivated in liquid medium. Efficient rooting (76%) was obtained using 16 µM IBA. 相似文献