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1.
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
2.
Vegetative propagation techniques are recognized as indispensable tools for mass multiplication of important multipurpose
trees adopted in different agroforestry systems. Albizia procera, one among important species, is difficult to propagate commercially either by stem / root cuttings or layering. A study
was undertaken to develop procedure for its in vitro regeneration through organogenesis. Explants collected from 15±2 yr-old mature plus trees and from 15 days old juvenile seedlings
were regenerated with exogenous application of different hormones. Epicotyl and hypocotyl explants excised from juvenile seedlings
showed higher callusing than axillary bud and shoot tip explants derived from mature trees. Benzylaminopurine (BA) at 3 μg/l
was most effective, which induced hundred percent callusing in epicotyl and hypocotyl explants in 1/2 Murashige and Skoog
(MS) medium. Callus originated from axillary buds and apical shoot tips of mature trees failed to form organs, however callus
derived from epicotyl and hypocotyl explants proliferated and formed de novo shoots and leaflets. A concentration of 3 μg/l of BA was found effective for shoot proliferation. Shoots grew vigorously
in 2 μg/l gibberellic acid (GA3) treatment and rooted in 1/2 MS medium supplemented with indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA). Rooting
was most successful on medium supplemented with 6 μg/l IBA alone on which 93.3% of the shoots formed roots. Sand or vermiculite
supplemented with 4 ml of yoshida solution proved as best hardening media, which recorded 70-80% survival of plantlets. One
year old tissue culture raised plants had comparatively more height, collar diameter, biomass, and root shoot ratio than plants
raised from cuttings and seeds of the same age. The procedures enumerated provide a basis for the development of in vitro techniques for rapid multiplication of A. procera.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
3.
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
4.
M. S. Shekhawat N. Kannan M. Manokari M. P. Ramanujam 《Journal of Sustainable Forestry》2013,32(4):327-336
A highly efficient, stable, and cost-effective micropropagation protocol for the conservation of a medicinal plant Turnera ulmifolia L. was established from nodal tissues via multiple axillary shoot proliferation on using Murashige and Skoog’s (MS) liquid nutrient medium. To begin with, nodal explants were placed on agar gelled medium amended with 2.0 mg L?1 6-benzylaminopurine (BAP) and 0.1 mg L?1 indole-3 acetic acid (IAA) for shoot induction. Subsequently, elongation of regenerated shoots could be possible on liquid MS medium supplemented with 0.5 mg L?1 BAP and Kin (kinetin) each along with 0.1 mg L?1 IAA where high frequency of regeneration in terms of number of shoots (47.2 shoots/explant) was achieved. Furthermore, long and healthy shoots (4?5 cm in length) were rooted on agar gelled half-strength of MS medium supplemented with 2.0 mg L?1 indole-3 butyric acid (IBA). Finally, in vitro regenerated plantlets were gradually acclimatized in the greenhouse and transferred to the field successfully. 相似文献
5.
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
6.
Mangal Singh M. S. Rathore D. Panwar J. S. Rathore H. R. Dagla 《Journal of Sustainable Forestry》2013,32(8):935-950
Aloe vera Linn. (Syn. Aloe barbadensis Mill; Gwar-patha in Hindi) belongs to family Liliaceae. The plant, for its medicinal properties, has commercial value. Some of the genotypes of Aloe vera are consumed as a vegetable and processed to make curry and other edible products. We report here on the development of an efficient method for rapid clonal propagation by shoot proliferation from axillary meristem(s) of selected germplasm of Aloe vera. Explants were pretreated with 0.1% aqueous solution of both streptomycin and bavistin separately, each for 15 min. These were surface sterilized with 0.1% aqueous solution of mercuric chloride (HgCl2) for 4–5 min and washed several times with autoclaved water. These were kept in a chilled, sterile antioxidant (200.0 mg L?1 of ascorbic acid, 50.0 mg L?1 of citric acid, and 25.0 mg L?1 of polyvinylpyrrolidone; PVP) solution and cultured on semi-solid Murashige and Skoog's (MS) medium. The bud explants produced multiple (10.3 ± 0.675/explant) shoots on MS medium containing 13.32 μM of 6-Benzylaminopurine (BAP) and 100.0 mg L?1 of ascorbic acid, 50.0 mg L?1 each of citric acid and PVP, with 25.0 mg L?1 each of arginine and adenine sulphate as additives. The shoots were further multiplied by (a) repeated transfer to fresh MS medium with additives + 13.32 μM BAP, and (b) subculturing on MS medium with a lower (4.44 μM) concentration of BAP. On MS medium containing 4.44 μM of BAP and additives, a maximum number (27.8 ± 0.63) of shoots were produced. In liquid MS medium with 4.44 μM of BAP, the rate of shoot multiplication increased and the vigor of the shoots improved. One hundred percent of the cloned shoots rooted under in vitro conditions on hormone-free half-strength MS salts containing 200.0 mg L?1 of activated charcoal at 32 ± 2°C. The cloned shoots treated with 2.46 mM of indole-3-butyric acid (IBA) or 2.473 mM of β-naphthoxyacetic acid (NOA) for 5 min rooted under ex vitro conditions in the greenhouse. The rooted plants were hardened in the greenhouse and stored under an agro-net house. The cloned plants were transferred under different field conditions at various sites in Western Rajasthan. These plants grew normally. The higher rate of shoot multiplication and easier approach of direct rooting and hardening make this method superior to the methods previously reported on cloning/tissue culture of Aloe species. From a single shoot bud, approximately 5000 plants can be produced within 180 days. 相似文献
7.
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
8.
An efficient protocol has been developed for in vitro propagation of Enicostema axillare using shoot tip explants. The shoot tip explants were cultured on MS medium supplemented with various combinations of (BAP, KIN) and (NAA/IAA & IBA) in different concentrations between 0.5 and 2.0 mg/l for multiple shoot bud induction. The highest percent of (98.51 %) was observed at 1.0 mg/l BAP in combination with 0.2 mg/l KIN while maximum number of shoot buds (8.41 shoots/explant) was noticed on MS medium containing 1.0 mg/l BAP and 0.2 mg/l KIN combination. The highest frequency (90.82 %) of multiple shoot bud regeneration was observed at 1.0 mg/l BAP and 0.5 mg/l IBA with 15.12 ± 2.12 shoots/explants. The regenerated multiple shoots were transferred to half-strength MS medium augmented with different concentration of 0.5–2.5 mg/l IBA for rooting. Among the different concentrations of IBA tested, maximum percentage of rooting (100 %) was observed in MS medium augmented with 1.5 mg/l IBA. The rooted plantlets were successfully transferred into plastic cups containing soil and sand in the ratio of 1:1. Subsequently established in the field conditions with 90 % of survival rate. The protocol developed can be utilized for both large scale plant production and conservation of germplasm of this species. The described method can be successfully employed for large-scale multiplication and in vitro conservation as well as production of secondary metabolites of E. axillare. 相似文献
9.
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
10.
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
11.
几种观赏竹种组织培养研究 总被引：15，自引：1，他引：15
以金镶玉竹等 11种观赏竹为试验材料 ,采用新萌发的嫩芽 ,在 MS BA 1.0 mg/ L (单位下同 ) IBA0 .1;MS BA1.0 KT0 .1 IBA0 .0 1;MS KT1.0 TDZ0 .0 0 1;MS BA5 .0 KT0 .1 IBA0 .0 1;MS BA1.0 KT0 .5培养基上成功获得 7种观赏竹的再生植株 ,丛生芽每2 0～ 4 0天继代 1次 ,不定芽增殖达 3～ 6倍。无菌苗移载成活率达 90 %以上。 相似文献
12.
《林业研究》2017,(1)
We developed a shoot multiplication protocol for Syringa reticulata Blume var. mandshurica Hara from in vitro cultured seedlings that derived from in vitro germinated seeds. The shoots could be induced on Murashige and Skoog(MS) medium with proper plant growth regulator combinations of 6-benzylaminopurine(BA) and indole-3-butyric acid(IBA). The better medium for shoot multiplication and growth was MS + 5 mg L~(-1)BA +0.5 mg L~(-1)IBA + 20 g L~(-1)sucrose +7 g L~(-1)agar, and the corresponding shoot induction rate was 75 %. The plantlets grew well after rooting on 1/2MS medium(macro-elements of MS medium are at half-strength) supplemented with 1 mg L~(-1)IBA, and the survival percentage was 80 % at 16 weeks after transplanting. 相似文献
13.
14.
《Journal of Sustainable Forestry》2013,32(1-2):159-170
Abstract Multiple shoots and plantlets were developed in vitro from cotyledonary nodal segments of in vitro raised seedlings of Anogeissus rotundifolia (syn. A. sericea var. nummularia)-a rare and endemic tree species of the Thar Desert. About 15-20 shoots differentiated from a single cotyledonary node within four weeks on Mu-rashige and Skoog's (MS) basal medium containing 0.1 mg l?1 indole-3-acetic acid (IAA) + 2.0 mg l?1 6-benzylaminopurine (BAP) + additives (25 mg l?1 each of adenine sulphate, L-arginine, and citric acid, and 50 mg l?1 of ascorbic acid) at 26 ± 2°C temperature and 36 μmol m?2 s?1 photon flux density with a 12 h/day photoperiod. The shoots produced in vitro were further multiplied by subculturing on fresh medium. The original cotyledonary nodal segment was repeatedly transferred (5 to 6 times) onto fresh medium containing 1.0 mg l ?1 BAP + 0.1 mg I ?1 IAA + additives to yield fresh crops of multiple shoots. These shoots were rooted on half-strength MS medium supplemented with 0.1 mg l?1 indole-3-butyric acid (IBA). Plantlets were transferred to pots containing sand-dune soil and ver-miculite it the ratio of 4:1 (v/v) and hardened in a growth chamber for two weeks and finally transferred to a greenhouse. From a single cotyledonary node about 1500 plantlets could be developed within four months. The method developed is useful for mass multiplication and for the conservation of germplasm of Anogeissus rotundifolia. 相似文献
15.
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
16.
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
17.
红掌的组织培养与快速繁育 总被引：3，自引：0，他引：3
选生长健壮的红掌盆苗,研究其组培快繁技术,结果表明：MS＋BA 1 mg/L＋NAA 0.2 mg/L＋葡萄糖30 g/L诱导率最高;1/2MS＋BA 2 mg/L＋2,4-D 0.5 mg/L＋椰汁100 mL/L＋蔗糖30 g/L培养基愈伤组织及芽苗增殖率最高;壮苗最佳培养基MS＋BA 0.5 mg/L＋IBA 0.05 mg/L＋椰汁100 mL/L＋蔗糖30 g/L;最佳生根培养基1/2MS＋NAA 0.3 mg/L＋蔗糖20 g/L,生根率达95%以上,当生根苗有3条-4条时即可移栽。 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.
Three superior clones of Eucalyptus grandis hybrids were micropropagated through several steps. Five-year-old trees were girdled to induce juvenile sprouts. Cultures were attempted from mature branches and sprouts. Branches from mature trees were 100% contaminated while sprouts were only 40% contaminated. Pre-initiation hormone free medium and dark environment were used to screen for contaminants and to reduce production of phenolic compounds. Initiation of auxillary buds was achieved with modified MS plus 0.05 mg/l NAA and 0.5 mg/l BAR High multiplication rates were obtained on auxin-free medium with 0.6 mg/1 BAR Elongation of shoots was best on media with high auxin (2.5 mg/l of IBA) and cytokinin (1–1.5 mg/l of zeatin). Continual subculture on the multiplication medium improved rooting significantly. Up to 98% rooting was achieved on 1/4 MS with 2 mg/l IBA. Rooted propagules were successfully transferred to a mist greenhouse with 82% survival, and then to greenhouse conditions. 相似文献