共查询到20条相似文献，搜索用时 31 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.
Cassiana de Oliveira Juliana Degenhardt-Goldbach Gisela Manuela de França Bettencourt Erika Amano Luziane Franciscon Marguerite Quoirin 《林业研究》2017,28(1):29-39
Genetic transformation systems require protocols that allow regenerating transgenic plants from transformed tissues. This study aimed to establish a protocol for indirect organogenesis in leaf explants of a Eucalyptus grandis × E. urophylla AEC 224 clone. During callogenesis stage, several concentrations of NAA and then NAA or 2,4-D combined with TDZ were tested in JADS culture medium for 30 days, followed by subculture of the explants in the regeneration medium, containing 5.0 µM BA and 0.5 µM NAA for another 30 days. In these media, the explant oxidation rate was high (95 %). Thus, in order to reduce oxidation, different culture media were compared: WPM, MS, JADS and modified QL, followed by explant transfer onto regeneration medium. The highest percentage of regeneration and the lowest oxidation rate were achieved on WPM medium. Then, NAA and 2,4-D were tested in combination with TDZ and also TDZ and BA combined with NAA in WPM medium. The most efficient culture media in terms of shoot regeneration were WPM supplemented with 0.25 µM TDZ and 0.1 µM NAA during 30 days for callus induction and then with 5.0 µM BA and 0.5 µM NAA for another 30 days. This protocol yielded a regeneration rate of 43 %, with a low oxidation of tissues. A rooting experiment was conducted using half strength MS medium and comparing three concentrations of IBA (2.46, 4.90 and 7.35 µM). The highest rooting percentage (35 %) was obtained on medium containing 2.46 µM IBA. Once the shoots were rooted, acclimatization in a greenhouse was not challenging and plant survival reached 100 %. 相似文献
3.
《林业研究》2016,(3)
The in vitro adventitious shoot differentiation in leaflet explants of an adult tree differed from that of leaflet explants of seedlings of Albizia procera(Roxb.)Benth. reported previously elsewhere. The leaflet explants from an adult tree passed through an initial callus phase for30 days on MS medium supplemented with 3 % sucrose,2.5 l M 2,4-D followed by a subsequent adventitious shoot differentiation phase for another 30 days on half MS medium supplemented with 0.25 l M each of BA and IBA.The regeneration rate of in vitro adventitious shoots in explants from the adult tree, i.e.1.66 shoots/callus, was lower than that from seedlings, i.e. [10 shoots/callus,which was reported elsewhere. Correspondingly, the activities of nitrate reductase and peroxidase, and endogenous phenol content remained very low during in vitro adventitious shoot differentiation in leaflet explants of an adult tree possibly due to lower availability of competent stem(juvenile) cells for the process. 相似文献
4.
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
5.
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
6.
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
7.
《林业研究》2019,(6)
This research reports on an efficient shoot proliferation and callus regeneration system for bamboo.Young, semi-lignified branches with one lateral bud from Drepanostachyum luodianense(Yi et R. S. Wang) Keng f.were used as explants. Disinfection with 0.1% HgCl_2 for 8 min was the optimum treatment and the best medium for bud initiation was Murashige and Skoog(MS) medium containing 3.0 mg L~(-1)6-benzyladenine(BA). Multiple shoots were induced from nodal shoot segments on MS medium containing 5.0 mg L~(-1) BA, 0.5 mg L~(-1) kinetin(Kin), and 1.0 mg L~(-1) naphthaleneacetic acid(NAA). The highest frequency of callus formation(65.6%) was on MS medium containing 4.0 mg L~(-1)2,4-dichlorophenoxyacetic acid(2, 4-D), 0.5 mg L~(-1) NAA, and 0.1 mg L~(-1) thidiazuron(TDZ). The optimum medium for callus proliferation was MS medium with 4 mg L~(-1)2,4-D, 0.5 mg L~(-1) TDZ and 0.5 mg L~(-1) NAA, and the optimum hormone combination was 4 mg L~(-1) BA ? 0.5 mg L~(-1) NAA for callus redifferentiation(up to 85.6%). A 100% rooting was achieved on MS medium supplemented with 2.0 mg L~(-1) NAA and 0.5 mg L~(-1)3-indole butyric acid(IBA). Rooted plantlets were acclimatized in a greenhouse in humus soil ? perlite(1:1) substrate. These micropropagated callus induction and regeneration systems for bamboo will be useful for genetic engineering and multiplication. 相似文献
8.
9.
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
10.
11.
唐巍 《中国林学(英文版)》2001,3(2)
三个基固型的火炬松成熟合子胚被培养在附加 8mg·L-12 ,4 D ,4mg·L-1BA ,4mg·L-1KT ,5 0 0mg·L-1水解酪蛋白和 5 0 0mg·L-1谷氨酰胺的愈伤组织诱导培养基上诱导愈伤组织 .来自于子叶、胚轴和胚根的愈伤组织在附加 1 6mg·L-12 ,4 D ,0 8mg·L-1BA和 0 8mg·L-1KT的愈伤组织增殖培养基上培养 9周后 ,可获得 16 9%的胚性愈伤组织 .通过建立胚性细胞悬浮系和研究ABA、PEG和活性炭对体细胞胚成熟的促进作用 ,优化的体细胞胚胎发生体系被建立 .71棵再生小苗被用于移栽试验 ,2 3棵小苗在田间移栽成活 相似文献
12.
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
13.
In vitro somatic embryogenesis in callus cultures of Azadirachta indica A. Juss.—a multipurpose tree
Gyana Ranjan Rout 《Journal of Forest Research》2005,10(4):263-267
Plant regeneration via somatic embryogenesis was achieved in embryogenic callus cultures derived from immature zygotic embryos 40 days after anthesis of Azadirachta indica A. Juss. on semisolid basal Murashige and Skoog (MS) salts and vitamins supplemented with 1.11 µM 6-benzylaminopurine (BA) and 4.52–6.78 µM 2,4-dichlorophenoxyacetic acid (2,4-D). The globular-stage embryos were induced when the callus was transferred to medium with 1.11 µM BA and 0.45 µM 2,4-D. The highest average number of somatic embryos per 200 mg of callus was 152.8 after 8 weeks of culture on the medium. Maturation and germination of the somatic embryos were achieved on half-strength MS salts and vitamins supplemented 0.38–0.94 µM abscisic acid (ABA) and 2% (w/v) sucrose. The maximum percentage (64.2%) of germination was obtained with 0.94 µM ABA within 2 weeks of culture. Somatic embryo-derived plantlets were acclimatized in a greenhouse and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones on a mass scale and will also be useful for genetic transformation study. 相似文献
14.
以MS为基本培养基，添加不同浓度的2，4-D、NAA、KT或其组合，研究库拉索芦荟愈伤组织的诱导及生长情况。结果表明，两种生长素均可诱导出愈伤组织，但较高浓度的2，4-D易导致褐化，而高浓度的NAA诱导出的愈伤组织含水量较高，以两种生长素配合使用再辅以低浓度的KT效果较好，其中15mg/L的KT 4mg/L的2，4-D 0．1mg/L的KT的组合，愈伤组织诱导率达到90％，且愈伤组织生长较旺盛。 相似文献
15.
16.
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 mg•L–1 BA plus 0.2 mg•L-1 NAA. In the presence of 0.2 mg•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
17.
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
18.
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
19.
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
20.
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