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1.
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
2.
Sanjaya Bagyalakshmi Muthan Thrilok Singh Rathore Vittal Ravishankar Rai 《Journal of Forest Research》2006,11(3):203-209
Santalum album is known as East Indian sandalwood. It is the most economically important tree harvested for heartwood oil, and India is
among the chief exporters of sandalwood and its products. Multiple shoots were induced from nodal shoot segments derived from
a 50- to 60-year-old candidate plus tree (CPT) on Murashige and Skoog (MS) medium supplemented with 0.53 μM α-naphthaleneacetic acid (NAA) and 11.09 μM 6-benzylaminopurine (BA). In vitro differentiated shoots were multiplied on MS medium with 0.53 μM NAA, 4.44 μM BA, and additives: 283.93 μM ascorbic acid, 118.10 μM citric acid, 104.04 μM cystine, 342.24 μM glutamine, and 10% (v/v) coconut milk. New shoots were harvested repeatedly for up to three subculture passages on fresh
medium at 4-week intervals. Microshoots treated with 98.4 μM indole-3-butyric acid (IBA) for 48 h produced roots on growth-regulator-free, quarter-strength MS basal salts medium with
vitamin B5 and 2% sucrose. In vitro root induction was achieved from microshoots pulsed with 1230 μM IBA for 30 min in soilrite rooting medium. The percentage of rooting in soilrite was higher than that for agar medium, and
in vitro raised plants were established in the field and showed normal growth. 相似文献
3.
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
4.
A.V. Raghu S.P. Geetha Gerald Martin Indira Balachandran P.N. Ravindran 《Journal of Forest Research》2006,11(1):57-60
An efficient system was developed for direct plant regeneration from in vitro-derived leaf explants of Embelia ribes Burm. f., a vulnerable medicinal woody climber of the Western Ghats of India. The in vitro procedure involved three steps
that included induction of shoot initials from leaf tissue, regeneration and elongation of shoots from the shoot initials,
and rooting of shoots. The induction of shoot initials was achieved on Murashige and Skoog (MS) solid medium supplemented
with different concentrations of thidiazuron (TDZ). The best medium for shoot induction was MS with 0.272 μM TDZ. Numerous shoot primordia developed within 2–3 weeks on the leaf margin as well as on the midrib region, without any
callus phase. In the second step, the shoot clumps separated from the leaf explant on transfer to MS basal medium, resulting
in the differentiation of 90% of the shoot initials into well-developed shoots. The 2- to 3-cm-long shoots rooted on half-strength
MS basal medium supplemented with 4.90 μM indole-3-butyric acid (IBA) and 3% (w/v) sucrose in the third stage. The rooted plants could be established in soil with
70% success. This protocol could be utilized for in vitro propagation and conservation of this important threatened medicinal
plant. 相似文献
5.
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
6.
《Journal of Sustainable Forestry》2013,32(2-3):103-114
Abstract Micropropagation protocols for Dendrocalamus asper using nodal shoots and seeds culture are described. Multiple shoots were induced through forced axillary branching. Ninety-five percent of the nodal shoot explants taken from juvenile primary and lateral branches, produced multiple shoots through axillary buds activation within 2 to 8 weeks on Murashige & Skoog's (MS) medium supplemented with 0.1-15 mg/l benzyladenine (BA). The cultured seeds also produced multiple shoots (1-20) within 6 weeks on this medium. The multiple shoot differentiation was influenced by the concentration of BA in the medium. The in vitro generated shoots were excised and subculture on MS + 3.0 mg/l BAP for further shoot multiplication. Fifteen to 20 fold rate of shoot multiplication was achieved by regular subculturing. These shoots were multiplied for more than 3 years without loss of vigor. Ninety-five percent of the shoots were rooted, when propagules (each consisting of cluster of 3 shoots) were transferred on to MS medium with 3.0 mg/l NAA or 10 mg/l indole-3-butyric acid (IBA). To date, 18,000 plants (through axillary bud initiated from nodal ex-plants) and 6,000 plants from seed culture have been hardened and acclimatized. 12,000 plants have been field transferred. 相似文献
7.
Gerald Martin S. P. Geetha Sudhakar S. Raja A. V. Raghu Indira Balachandran P. N. Ravindran 《Journal of Forest Research》2006,11(6):461-465
A micropropagation protocol was developed for Celastrus paniculatus, a vulnerable medicinal plant. Cultures were initiated from nodal explants collected from young shoots of a 12-year-old plant
in MS basal medium. An average of five shoots were produced in MS medium supplemented with 1.5 mg l−1 benzyl adenine (BA) and 0.1 mg l−1 naphthalene acetic acid (NAA) after two subculture cycles with a 30-day interval. Continuous subculture in the same medium
for three more cycles resulted in reduction of the number of multiple shoots (2 or 3 shoots), vitrification of the shoots,
and callus formation. Vitrification of cultures could be overcome by the use of MS medium supplemented with lower concentrations
of BA (0.05 mg l−1) and NAA (0.01 mg l−1). Among the various rooting trials, ex vitro rooting of shoots with simultaneous hardening was most efficient. The method
standardized in the present study is simple, as it eliminated separate steps for in vitro rooting and hardening. Qualitative
chemical similarity of the tissue culture regenerants with the mother plant was confirmed using high performance thin-layer
chromatographic (HPTLC) profiling. 相似文献
8.
This report describes a protocol for the in vitro shoot induction and plant regeneration from epicotyl explants of Cassia angustifolia on Murashige and Skoog (MS) medium supplemented with 6-benzyladenine (BA), Kinetin and 2-iP (0.5–10.0 μM). MS medium supplemented with BA (5.0 μM) was the most effective in inducing adventitious shoots and growth. The highest rate of shoot multiplication was achieved on MS medium supplemented with BA (5.0 μM) and Indole-3-acetic acid (IAA, 1.0 μM). The nodal segments excised from the shoots regenerated from BA (5.0 μM) and IAA (1.0 μM) were used as explants for next three round of micropropagation. The number of shoots significantly increased at successive round of micropropagation. For rooting, MS medium supplemented with 2.0 μM indole-3-butyric acid proved to be better than that supplemented with IAA or α-naphthalene acetic acid. The in vitro raised plantlets with well developed shoot and roots were successfully established in earthen pots containing garden soil and were grown in greenhouse. About 52 plants (85 %) survived out of 60 plants transferred in garden soil. 相似文献
9.
Nothofagus leoni has a restricted distribution in Chilean forests. This work determines suitable culture conditions forin vitro multiplication and rooting through shoots obtained from seedlings. Broadleaved Tree Medium was suitable for shoot multiplication.
A medium with a pulse of 0.55 μM BA in the first subculture and two subcultures on BA-free medium resulted in a multiplication
rate at day 63 of × 5.7, without callus growth or shoot neoformation. Rooted shoots of good quality (number and length of
roots without callus growth) were obtained with 1.23 μM IBA (91.4% of rooting). The first roots appeared at day 11, reaching
a higher speed of rooting at day 15. 相似文献
10.
A method for rapid in vitro propagation of Cassia siamea Lam. using cotyledonary node explants, excised from 14-day old aseptic seedlings, has been established. Murashige and Skoog
(MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn) and thidiazuron (TDZ) singly
or in combination with auxins was used for regeneration studies. Among the single treatment of three cytokinins BA at 1.0 μM
was found to be optimum for direct shoot regeneration as it induced an average of 8.20 ± 0.66 shoots per explant. The regeneration
frequency further enhanced with the application of auxin along with optimal BA concentration. The highest frequency for shoot
regeneration (90%), the maximum number of shoots per explant (12.20 ± 0.73) and the maximum shoot length (6.40 ± 0.07) cm
were obtained on the medium consisted of MS + 1.0 μM BA + 0.5 μM NAA. Successful in vitro rooting was induced from cut end
of the microshoots when placed on half-strength MS + IBA (2.5 μM). The regenerated shoots with well developed root system
were successfully acclimatized and established in pots containing sterilized garden soil and garden manure (1:1) and grown
under greenhouse conditions with 85% survival rate. 相似文献
11.
Mahendra Phulwaria Kheta Ram Harish Amit K. Gupta N. S. Shekhawat 《Journal of Forest Research》2012,17(2):202-207
We report an efficient in vitro propagation method for Terminalia catappa using nodal segments of a 15-year-old mature tree. The nodal segments were cultured on MS medium supplemented with 6-benzyladenine
(BA; 0.5–3.0 mg l−1) or Kinetin (Kn; 0.5–3.0 mg l−1) for bud breaking and multiple shoot induction. About 85% of the explant responded (2.8 ± 0.41 shoots per node with 2.7 ± 0.14 cm
length) within 15 days of inoculation in Murashige and Skoog medium fortified with 2.0 mg l−1 of BA. Further shoot multiplication was achieved by repeated transfer of mother explants and subculturing of in vitro-produced
shoots on medium supplemented with various concentrations of BA (0.25–1.5 mg l−1) or Kn (0.25–1.5 mg l−1) or on their combinations. Optimal number of shoots and shoot length were recorded on MS medium supplemented with 0.25 mg l−1 of BA and 0.25 mg l−1 of Kn. The multiplied shoots were used for ex vitro rooting after treatment for 4 min with indole-3-butyric acid (IBA; 50–500 mg l−1) or α-naphthalene acetic acid (NAA; 50–500 mg l−1). About 80% of the shoots treated with 200 mg l−1 of IBA produced ex vitro roots with an average of 2.8 roots per shoot. Nearly 75% of these plantlets could be acclimatized
within 5 weeks and successfully established in the field. This is the first report on micropropagation of T. catappa, which can be applied for further genetic transformation assays and pharmaceutical purposes. 相似文献
12.
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
13.
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
14.
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
15.
Bi-huạ Chen 《中国林学(英文版)》2009,11(3):174-178
In this study a reliable protocol was developed for the establishment of commercial in vitro cultures of Tripterygium wilfordii Hook f.. Juvenile shoots from one-year-old elite plants were used as the source of explants. New axillary shoots were obtained after 30 days of culture on a MS medium supplemented with BAP (2.0 mg·L–1) and NAA (0.1 mg·L–1). The optimal multiplication medium was a modified MS medium supplemented with BAP (1.0 mg·L–1) and NAA (0.1 mg·L–1). This yielded a multiplication rate of 2.4 for each subculture. Slightly more than 92% of shoots rooted when cultured on a modified MS medium containing IBA (0.2 mg·L–1) and acti-vated charcoal (0.5 mg·L–1). Activated charcoal promoted both a strong and a high rooting rate during the rooting phase. Plantlets were transferred to pots for a short acclimatization stage in a greenhouse where 95% of the plantlets survived. This highly reproduci-ble procedure can be adopted for large-scale propagation of T. wilfordii. 相似文献
16.
A protocol for micropropagation using nodal explants from mature Pinus massoniana trees has been developed. Time of explant collection is crucial for the initial success of aseptic culture. Explants collected in early March gave the highest percentage of explant survival (64.5%) and shoot-forming percentage (52.3%). Thidiazuron (TDZ) concentration significantly influenced shoot formation; 4 μM TDZ was optimum, with 4.8 shoots produced per explant with a mean length of 7.1 cm after 120 days of culture. Regenerated shoots rooted for 60 days in basic medium with 1 μM NAA were ready for growth in pots. This is the first report on plantlet regeneration in vitro from mature trees of P. massoniana that provides a reliable method for propagating selected elites. 相似文献
17.
Experiments were conducted to study plant regeneration through direct somatic embryogenesis using mature zygotic embryo and
cotyledonary explants from seeds of Melia volkensii stored for <3 and >12 months. Explants were cultured on Murashige and Skoog (MS) medium supplemented with BAP, NAA and 2,4-D
(0.5, 1.0 and 2.0 mg l−1) alone, and BAP (0.5, 1.0, 2.0 and 4.0 mg l−1) in combination with 2,4-D or NAA (0.2 and 0.5 mg l−1). After 4 weeks in culture, up to 60% of cotyledonary explants from the seeds stored for <3 months produced direct somatic
embryos on BAP (0.5–4.0 mg l−1) in combination with 2,4-D (0.2 mg l−1). The number of somatic embryos ranged from 5 to 14 per explant in BAP (0.5 mg l−1) and 2,4-D (0.2 mg l−1) combination. Only 20% of cotyledonary explants from seeds stored for >12 months produced somatic embryos. Mature zygotic
embryos failed to produce any somatic embryos. Subcultures of somatic embryos from cotyledonary explants of seeds stored for
<3 months formed clusters of shootlets on semi solid MS and 1/2 MS media. After 6 weeks of subculture on multiplication MS
media augmented with BAP (0.5 mg l−1) and IAA (0.2 mg l−1), 70% of the shoot tips formed 4–7 shoots per explant. Up to 33% of the multiplied shoots were rooted in MS medium supplemented
with 2.0 mg l−1 IBA. Plantlets developed normally into seedlings in the greenhouse. 相似文献
18.
An efficient method for cultivation of Vitex negundo L. through axillary shoot (collected from micropropagated plants) proliferation has been successfully developed, which can be employed at a commercial scale. Axillary shoot induction was most successful using nodal explants for propagation on Murashige and Skoog (MS) medium supplemented with various concentrations of single cytokinin or in various combination with auxins. We obtained the maximum percentage (97.6 ± 1.45) response with highest number (16.4 ± 0.60) of shoots per explant on MS medium supplemented with 6-benzyladenine (BA) and ??-naphthalene acetic acid (NAA) at concentrations of 5.0 and 0.5 ??M, respectively. Shoot regeneration frequency was optimized by manipulating pH and using various media. MS medium and pH 5.8 was found to be the optimum for maximum regeneration. Nodal explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. Shootlets with 4?C5 nodes were utilized for in vitro rooting, and best response was evaluated on MS medium supplemented with 1.0 ??M indole-3-butyric acid (IBA). The well-developed micropropagated plants were acclimatized (97%) successfully within 2 weeks in soilrite and planted ex vitro in normal garden soil, where they grew well without any morphological and genetic variations. The present regeneration process not only favoured the rapid multiplication but also expressed the regeneration capability of micropropagated plants. 相似文献
19.
用取自1、5和10年生母本上的杉木茎段作为原始外植体,接种于含有BAP 0.75mg/l和IBA 0.25mg/l的“GN”培养基中(NH_4~+:NO_3~-=1∶2.86)。经1a连续4代培养,结果表明:嫩梢及苗木产量因外植体母本的年龄不同有很大差异。如母本年龄为1a的外植体,每个平均可获嫩梢170.3—445.7根,其中有效嫩梢78.0—194.9根,包括Ⅰ级(好的,可供继代培养继续增殖)11.1—25.1根;Ⅱ级(中等,可供生根培养以成苗)36.1—87.9根;Ⅲ级(须继续培养)29.5—78.6根。随着原始外植体母本年龄的增加,以上各项数值都相应下降。 另一方面,由于组织培养对于成年母本的外植体具有不同程度的复壮效应,使得嫩梢增殖效果差异很大。如取自年龄为5、10年生母本上的外植体,平均年嫩梢产量为4.1—234.3根,如此大范围的变化为培养体的复壮选择提供了可能性。 此外,本文还对嫩梢增殖的生产效应作了初步分析。 相似文献
20.
Rooting of shoots derived from axillary buds was examined to establish an efficient shoot culture system of clonal micropropagation
in adult tree ofLarix leptolepis Gord. (Japanese larch). Nine out of ten shoots induced calli (90%) on their shoot bases, and the two of them formed root
primordia with a red pigment (20%) on the calli surface within 5 weeks after culturing on modified Murashige and Skoog (MS)
medium supplemented with 1.5 μM of indolebutyric acid (IBA) and 1.5 μM of naphthaleneacetic acid (NAA). However, the primordia
did not elongate actively. The addition of 10 mMl-phenylalanine in the MS medium with the auxins resulted in the formation of roots at high frequency, about 80%, and they
elongated actively. Although callus was formed in all the shoots cultured on the medium withl-phenylalanine, it appeared that the callus development was less as compared to the medium withoutl-phenylalanine. Consequently, the rooting might be associated with the suppression of the induced callus. 相似文献