共查询到20条相似文献，搜索用时 174 毫秒
1.
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
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.
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
4.
We developed procedures for the micropropagation of Juniperus oxycedrus L. using shoot apices or nodal segments from mature plants. Of the media and explants examined, best culture establishment was obtained with shoot apices cultured on modified Schenk and Hildebrandt medium (SH medium) without growth regulators; however, shoot multiplication was only achieved when shoot apices isolated from shoots grown on SH medium without growth regulators were subcultured on SH medium containing 0.5 micro M benzyladenine. None of the auxins and methods tested for root induction provided satisfactory results. 相似文献
5.
A comparative performance of two different media formulations (woody plant medium (WPM) and Murashige and Skoog??s (MS) medium) for their ability to inflict in vitro shoot development in nodal segments of Salix tetrasperma Roxb. has been carried out. Thidiazuron (TDZ) in various concentrations was used as a supplement to the basal media. Media types, TDZ concentrations, exposure duration and culture regimes played an important role in affecting multiple shoot production. WPM supplemented with 2.5???M TDZ for 4?weeks exposure was found to be the best for maximum (4.53?±?0.27) shoots production in vitro. Transfer to a secondary medium consisting of 6-benzyladenine (1.0???M) and ??-naphthalene acetic acid (0.5???M) enhanced the multiplication rate and by the end of 12?weeks, 20.33?±?0.33 shoots with shoot length, 4.70?±?0.26?cm were produced on WPM. Rooting of the regenerated shoots was achieved on half strength basal media (either WPM or MS) containing 0.5???M indole-3-butyric acid. In all the experiments, different growth parameters were scored and WPM was found to be superior to MS medium. The regenerated plantlets were successfully acclimatized in the field with about 81?% survival. 相似文献
6.
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
7.
以3个非洲菊品种的叶柄为材料,研究了不同激素及其浓度对非洲菊离体培养再生的影响。结果表明:3个品种的叶柄在含单一细胞分裂素6-BA的培养基上培养时都不能被诱导出愈伤组织;而在仅含生长素NAA的培养基上培养时均可被诱导出愈伤组织,并且在其愈伤组织发生部位都有不定根发生,但只有品种6267能从不定根发生部位直接分化出不定芽;当在含NAA的培养基上再附加6-BA时也可被诱导出愈伤组织,但无不定根发生。所产生的愈伤组织在分化培养基上培养时只有由品种Ⅱ叶柄在同时含有NAA和6-BA的诱导培养基上产生的愈伤组织才可以分化出不定芽。表明愈伤组织的诱导与分化、不定芽和不定根的发生与品种及培养基中的激素种类有关。 相似文献
8.
9.
A micropropagation protocol was established for a medicinal plant Vitex negundo. Genetic stability of micropropagated plants was investigated. Multiple shoots were induced from nodal explants cultured on Murashige and Skoog (MS) medium with 0.53 μM naphthalene acetic acid (NAA) and 11.0 μM benzyl aminopurine (BAP) along with additives (ascorbic acid, 283.9 μM; citric acid, 130.1 μM; and arginine, 143.6 μM). Shoots were further multiplied by repeated transfer of the mother explant. The shoots were further multiplied on MS medium + 0.57 μM indole-3-acetic acid (IAA) and 6.6 μM BAP. The micropropagated shoots were pulse treated with 122.5 μM indole-3-butyric acid (IBA), in liquid MS medium and then transferred to autoclaved soilrite. These rooted ex vitro. Shoots were also rooted in vitro on a half-strength MS medium + 2.45 μM IBA. The survival rate of in vitro rooted plantlets was poor during hardening compared to ex vitro rooted plantlets. About 95% of the ex vitro rooted, hardened plantlets survived in the field. Genetic stability of micropropagated plants was tested by using 25 random amplified polymorphic DNA primers. The cloned plants exhibited no variation in banding pattern in comparison with the mother plant. 相似文献
10.
Acacia sinuata is a valuable multipurpose tree in Southern India. The tree is over exploited, but its regeneration rate in natural habitat is low. Therefore, it is important to study if it can be regenerated through in vitro micro-propagation. Cotyledonary node and shoot-tip explants excised from 15 day-old in vitro grown seedlings were used to initiate cultures. Maximum number of shoots was induced from cotyledonary node explants on Murashige and Skoog's (MS) medium containing6.66 µM 6-benzylamino purine (BAP) and 4.65µM kinetin (Kn). Subculturing was done in the fresh medium of same composition. The number of shoots formed was comparatively greater in the first subculture. Maximum shoot elongation was achieved (5.5 cm)when subcultured on MS medium supplemented with 1.75 µMgibberellic acid (GA3). In vitro regenerated shoots produced roots when transferred to half strength MS medium supplemented with 7.36 µM indolebutyric acid (IBA). From each cotyledonarynode 30 shoots were obtained within 90 days after two subcultures. The success rate of establishing the rooted plantlets in the field was 55%.This revised version was published online in November 2005 with corrections to the Cover Date. 相似文献
11.
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
12.
Multiple shoots of high quality were produced in vitro from nodal expiants of Tectona grandis. An average of about 4 shoots/uninodal expiant was obtained within 4 weeks of culture on Murashige and Skoog’s (mMS) medium modified by 50% reduction in NH4NO3 concentration, supplemented with benzylaminopurine (1.5 mg L?1); indole-3-butyric acid (0.01 mg L?1) and gibberellic acid (0.1 mg L?1). The latter was applied both in the medium and by soaking the nodal segments for 10 s. in a gibberellic acid solution of 100 mg L?1. Hundred percent of shoots rooted cultured on modified MS medium containing IBA (0.5 mg L?1) and putrescine (160 mg L?1). Putrescine promoted both strong and highly ramified roots and fast growing shoots during the rooting phase, conditioning the plantlets for a good survival and quality. Plantlets were transferred to jiffy pots for a short acclimatization stage in greenhouse where they survived at 100%. This highly reproducible procedure can be adopted for large scale teak propagation. 相似文献
13.
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
14.
In vitro regeneration of complete plants from nodal single-bud segments of 2-year-old Australian Cedar (Toona ciliata) trees were obtained under defined nutritional and environmental conditions. Explants were dissected from plants obtained by germination of seeds and growth in pots in a greenhouse. The best medium for shoot regeneration was that of Murashige and Skoog at 1/4 strength with 3% sucrose (1/4 MS), supplemented with 0.1 mg/l IBA and 0.5 mg/l BAP. Rooting of regenerated shoots was observed in MS medium with 0.1 mg/l IBA. Using mature tree material was more difficult. Forced flushing was used to induce shoot development on branches of a 10-year-old tree. Nodal segments of these epicormic shoots formed shoots in vitro on 1/4 MS + 0.01 mg/l IBA + 5 mg/l BAP, but rooting was never observed. 相似文献
15.
《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
16.
Mangal S. Rathore S. Shekhawat G. Kaur R. P. Singh N. S. Shekhawat 《Journal of Sustainable Forestry》2013,32(8):727-746
Withania coagulans (Stocks) Dunal (Solanaceae), popularly called vegetable rennet, is a critically endangered and highly valued medicinal plant. Overexploitation and reproductive failure forced the plant species toward the verge of complete extinction. We describe here the development of a simple, rapid, and cost effective in vitro micropropagation system for W. coagulans for mass-scale production of true-to-type plantlets using nodal shoot segments. Exactly 95.5 ± 0.34% explants responded within 8–10 days (d) and produced multiple shoot buds (4.1 ± 0.10 shoots of 2.95 ± 0.15 cm length) on 0.8% agar-gelled Murashige and Skoog's (MS) basal medium supplemented with 8.88 μM 6-benzylaminopurine (BAP), 0.57 μM indole-3-acetic acid (IAA), and additives (100 mg L?1 L-ascorbic acid, 25 mg L?1 each citric acid, adenine sulphate, and L-arginine). The shoots in cultures were multiplied by repeated transfer on MS medium with 4.44 μM BAP, 0.57 μM IAA, and additives. Further cultures were multiplied on a large-scale through the subculturing of shoot clumps differentiated in vitro, on MS medium supplemented with 1.11 μM BAP, 0.57 μM IAA, and additives. Maximum number (19.1 ± 0.28) of healthy (6.15 ± 0.25 cm) and viable shoots differentiated on this medium. The microshoots were rooted both in vitro and ex vitro. Exactly 67.3 ± 1.01% microshoots rooted in vitro within 25–30 d on agar-gelled half-strength MS salts supplemented with 29.52 μM indole-3-butyric acid (IBA) and 200 mg L?1 of activated charcoal (AC). Alternatively, 73.8 ± 0.65% cloned shoots rooted on sterile soilrite (soilless compost and soil conditioner) under ex vitro conditions after pulse treatment with 2.46 mM IBA for 300 s. The clones of W. coagulans were hardened in a greenhouse within 40–45 d by slow and gradual exposure of plantlets from high relative humidity (RH; 70–80%) and low (26 ± 2°C) temperature to low RH (40–50%) and high (34 ± 2°C) temperature. The hardened plantlets were transferred to soil and stored in agro-net house with more than 90% survival rate. Replacement of pure and laboratory grade sucrose with commercial grade sugar, use of less expensive commercial grade agar-agar in culture medium, higher rate of shoot proliferation, single step ex vitro rooting, and hardening of plantlets in the greenhouse are advantageous features of the protocol. The micropropagation protocol defined here is reproducible, easy to follow, and would be helpful in large-scale restoration programs through true-to-type mass-multiplication of W. coagulans. 相似文献
17.
Callus cultures were established from internodal segments of shoot cultures from two mature black locust Robinia pseudoacacia L. trees. Callus of both trees produced shoots on Murashige and Skoog (MS) medium supplemented with 10 microM 6-benzylaminopurine alone or in combination with 1 microM naphthaleneacetic acid. Regenerated shoots were successfully multiplied on MS medium containing 0.32 microM 6-benzylaminopurine, and produced roots on 0.1 strength MS medium containing 1 microM indole-3-butyric acid. One clone consistently outperformed the other with respect to shoot proliferation and proportion of shoots that produced roots. This distinction had previously been observed in shoots produced from bud explants obtained from the mature trees. 相似文献
18.
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
19.
The objective of the study was to develop an in vitro shoot regeneration protocol by utilising shoot tips explant from Vitex trifolia L. Shoot tip explants obtained from a 3-year old plant was cultured on Murashige and Skoog (MS) medium supplemented with various concentrations (1.0, 2.5, 5.0, 7.5 or 10.0 µM) of thidiazuron (TDZ). The optimal level of TDZ supplementation to the culture medium was 5.0 μM for 15 d induction period. The highest number of shoots (22.2 ± 0.1) and shoot length (5.1 ± 0.1 cm) were achieved when TDZ-exposed explants were sub-cultured on MS medium containing 6-benzyladenine (1.0 µM) and 0.5 µM α-naphthalene acetic acid (NAA) after 8 weeks of culture. In vitro rooting of isolated shoots was achieved best in half-strength MS medium containing 0.5 µM NAA. During the acclimatization period, changes in activities of antioxidant enzymes were observed. Superoxide dismutase activity increased reaching maximum at 28th day after transplantation. Likewise, an upregulation of the catalase, ascorbate peroxidase and glutathione reductase enzyme activities were also observed. These observed changes reflected the ability of plants in developing an antioxidant enzymatic defence system aiding in survival against oxidative stress and in reducing release of free radicals. Plantlets were successfully hardened off and acclimatized in earthen pots containing garden soil with a survival rate of 90 %. 相似文献
20.
An in vitro regeneration system was developed using organogenic callus derived from in vitro grown cotyledonary explants of Gleditsia caspica Desf., an important leguminous tree. Murashige and Skoog (MS) basal medium augmented with 0.2 g L?1 myo-inositol and various concentrations of either 2,4-dichlorophenoxyacetic acid (2,4-D), naphthaleneacetic acid, or indole-3-butyric acid (IBA) alone as well as combined with cytokinins was used for callus induction. The highest frequency of organogenic yellowish-white and nodular callus (93 %) was obtained from explants grown on medium supplemented with 13.5 μM 2,4-D and 4.4 μM benzyladenine (BA). The yellowish-white and nodular callus when transferred to MS medium supplemented with BA (2.2–17.7 μM) or kinetin (KT; 2.3–18.8 μM) solely or in combination with 2.3 μM 2,4-D produced several microshoots after 5 weeks culture. The calli cultured on MS medium with 4.4 μM BA singly showed superior growth response and produced both maximum shoot regeneration (94 %) and the highest mean number (4.3) of microshoots per callus. Transfer of regenerated microshoots onto modified MS basal medium fortified with 5.8 μM gibberellic acid and 4.4 μM BA resulted in the maximum number of internodes per shoot and the highest shoot elongation after a period of 6 weeks. Optimum rooting of 90 %, an average 6.1 roots per shoot, and a mean root length of 3.6 cm was observed when half-strength MS medium was supplemented with 9.8 μM IBA and 0.92 μM KT. The regenerated healthy plants with well-developed shoots and roots showed a survival rate of 77 % after acclimatization and transplanting to garden soil for a 10-week hardening period under ex vitro conditions. 相似文献