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1.
This paper describes multiple shoot regeneration from leaf and nodal segments of a medicinally important herb Centella asiatica L. on Murashige and Skoog’s (MS) medium supplemented with a range of growth regulators. The highest number of multiple shoots
was observed on MS augmented with 3.0 mg dm−3 N6-benzylaminopurine (BAP) and 0.05 mg dm−3 α-naphthaleneacetic acid (NAA). Leaf explant showed maximum percentage of cultures regenerating shoots (81.6 %), with the
highest shoot number (8.3 shoots per explant) and the shoot length (2.1 cm) whereas, nodal explant showed less number of shoots
with callus formation at the base cut end. Successive shoot cultures were established by repeatedly sub-culturing the original
explant on a fresh medium. Rooting of in vitro raised shoots was best induced on half strength MS supplemented with 0.5 mg dm−3 indole-3-butyric acid (IBA) with highest percentage of shoot regenerating roots (76.8 %) with 3–4 roots per shoot. Plantlets
were acclimated in Vermi-compost and eventually established in soil. Contents of chlorophyll, total sugars, reducing sugars and proteins were estimated in
leaf tissue from both in vivo and in vitro raised plants. Chlorophyll content was higher in in vivo plants, whereas other three components were higher in in vitro plants. 相似文献
2.
Mingjia Yang Xiangming Xie Caixia Zheng Fangqiu Zhang Xiaoqing He Zhiru Li 《Plant Cell, Tissue and Organ Culture》2008,95(2):141-147
A protocol was developed for Agroacterium-mediated genetic transformation of Acacia crassicarpa via organogenesis by using in vitro phyllode (leaf) as the explant. Phyllode (leaf) explants were co-cultured with Agrobacterium tumefaciens strain LBA4404 harbouring binary vector pBI101 (harboring antisense Pt4CL1 with respect to the Pt4CL1P promoter). The selection for transgenic shoots was performed through two consecutive steps on
Murashige and Skoog (MS) medium supplemented with different concentrations of plant growth regulators and antibiotics in the
following order: 0.5 mg/l thidiazuron (TDZ), 0.5 mg/l α-naphthaleneacetic acid (NAA), 300 mg/l carbenicillin (Car) and 20 mg/l
kanamycin (Km) for 10 days; 0.1 mg/l TDZ, 200 mg/l Car and 20 mg/l Km for 60 days; 0.5 mg/l indole-3-butyric acid (IBA), 100 mg/l
Car and 20 mg/l Km 50 days. 21.7% of nodules produced multiple adventitious shoot buds, of which 27.7% survived in initial
selection. The shoot buds were subjected to repeated selection on MS medium supplemented with 0.1 mg/l TDZ, 200 mg/l Car and
20 mg/l Km for 60 days. Transgenic plants were obtained after rooting on half-strength MS medium supplemented with 0.5 mg/l
IBA, 100 mg/l Car 20 mg/l Km 50 days. Genomic PCR analysis confirmed the incorporation of the antisense Pt4CL1 with respect to the Pt4CL1P promoter fragment into the host genome. 相似文献
3.
Semiha Erisen Mustafa Yorgancilar Emine Atalay Mehmet Babaoglu 《Plant Cell, Tissue and Organ Culture》2010,100(2):229-233
Prolific shoot regeneration via organogenesis was induced from leaf and leaf petiole explants of the endemic Astragalus cariensis species on Murashige and Skoog (MS) medium with α-naphthaleneacetic acid (NAA) and benzyladenine (BA) within 8 week. The
highest number of shoots (23/explants) was obtained from leaf explants cultured on MS with 0.5 mg/l NAA and 4 mg/l BA. Elongated
shoots were successfully rooted in MS medium with 0.5 mg/l indole-3-butyric acid. Rooted plantlets were acclimatized in pots
containing 1:1 mixture of peat and perlite. 相似文献
4.
In vitro propagation of northern red oak (Quercus rubra) shoots was successful from cotyledonary node explants excised from 8-wk-old in vitro grown seedlings. Initially, four shoots per explant were obtained on Murashige and Skoog (MS) medium supplemented with 4.4 μM
6-benzylaminopurine (BA), 0.45 μM thidiazuron (TDZ), and 500 mg l−1 casein hydrolysate (CH) with a regeneration frequency of 64.7% after 3 wk. Subculturing explants (after harvesting shoots)
to fresh treatment medium significantly increased shoot bud regeneration (16.6 buds per explant), but the buds failed to develop
into shoots. A higher percentage (73.3%) of the explants regenerated four shoots per explant on woody plant medium (WPM) supplemented
with 4.4 μM BA, 0.29 μM gibberellic acid (GA3), and 500 mg l−1 CH after 3 wk. Explants subcultured to fresh treatment medium after harvesting shoots significantly increased shoot regeneration
(16 shoots per explant). Shoot elongation was achieved (4 cm) when shoots were excised and cultured on WPM supplemented with
0.44 μM BA and 0.29 μM GA3. In vitro regenerated shoots were rooted on WPM supplemented with 4.9 μM indole-3-butyric acid. A higher percentage regeneration response
and shoot numbers per explant were recorded on WPM supplemented with BA and GA3, than on MS medium containing BA and TDZ. Lower concentrations of BA and GA3 were required for shoot elongation and prevention of shoot tip necrosis. Each cotyledonary node yielded approximately 20
shoots within 12 wk. Rooted plantlets were successfully acclimatized. 相似文献
5.
Summary Tennessee coneflower [Echinacea tennesseensis (Beadle) Small] was regenerated from flower stalks, leaf sections from flowering plants, and hypocotyls and cotyledons from
seedlings. Murashige and Skoog medium (MS) supplemented with naphthaleneacetic acid (NAA) at 0.54 μM and thidiazuron (TDZ) at 22.7 μM yielded the most shoots per leaf explant. NAA and 6-benzylaminopurine concentrations for optimal shoot regeneration from
leaf, flower stalk, cotyledon and hypocotyl explants in MS media were 0.54 and 24.6μM, respectively. All explant types generated shoots; however, those derived from leaves and flower stalks produced the highest
number of shoots per explant and highest percentage of explants with shoots. Explants cultured on media containing high levels
of NAA (5.4–27 μM) formed calluses but no adventitious shoot. Leaf explants responded to a wider range of NAA concentrations than the other
explant types but shoots generated from flower stalks grew the fastest. While all cytokinins tested increased the number of
shoots per explant, the number of shoots in media containing TDZ was increased by nearly threefold. Regenerated shoots from
all explant types cultured on MS medium supplemented with 0.25 μM indole-3-butyric acid initiated roots within 4 wk; NAA was not effective for root induction. All vernalized plantlets developed
into plants that were morphologically identical to the source material. 相似文献
6.
Esther E. Uchendu Gopinadhan Paliyath Dan C. W. Brown Praveen K. Saxena 《In vitro cellular & developmental biology. Plant》2011,47(6):710-718
North American ginseng (NAG) (Panax quinquefolius L.) is a medicinally important plant with multiple uses in the natural health product industry. As seed propagation is time-consuming
because of the slow growth cycle of the plant, in vitro propagation using a bioreactor system was evaluated as an effective approach to accelerate plant production. An efficient
method was developed to multiply nodal explants of NAG using liquid-culture medium and a simple temporary immersion culture
vessel. The effects of plant growth regulators, phenolics, and chemical additives (activated charcoal, melatonin, polyvinylpolypyrrolidone,
and ascorbic acid) were evaluated on in vitro-grown NAG plants. The highest number (12) of shoots per single node was induced in half-strength Schenk and Hildebrandt basal
medium containing 2.5 mg/l kinetin, in which 81% of the cultured nodes responded. In a culture medium with 0.5 mg/l α-naphthalene
acetic acid (NAA), roots were induced in 78% of the explants compared to 50% with a medium containing indole-3-acetic acid.
All of the resulting plants appeared phenotypically normal, and 93% of the rooted plants were established in the greenhouse.
Phenolic production increased significantly (P < 0.05) over a 4-wk culture period with a negative impact on growth and proliferation. Activated charcoal (AC; 50 mg/l) significantly
reduced total phenolic content and was the most effective treatment for increasing shoot proliferation. Shoot production increased
as the phenolic content of the cultures decreased. The most effective treatment for NAG development from cultured nodal explants
in the bioreactor was 2.5 mg/l kinetin, 0.5 mg/l NAA, and 50 mg/l AC in liquid culture medium. This protocol may be useful
in providing NAG tissues or plants for a range of ginseng-based natural health products. 相似文献
7.
Guirong Qiao Jing Zhou Jing Jiang Yuehua Sun Luanyin Pan Honggai Song Jingmin Jiang Renying Zhuo Xiaojuan Wang Zongxiu Sun 《Plant Cell, Tissue and Organ Culture》2010,102(2):163-170
Malaxis acuminata is a terrestrial orchid that grows in shady areas of semi-evergreen to shrubby forests. It is highly valued for its medicinal
properties as dried pseudo-bulbs are important ingredients of several Ayurvedic preparations. In this study, adventitious
shoot buds were induced from internodal explants of M. acuminata grown on Murashige and Skoog (MS) medium supplemented with different concentrations of 6-benzyladenine (BA), kinetin (Kn),
and thidiazuron (TDZ). Of the three cytokinins used, TDZ at 3 mg l−1 induced the highest frequency (82%) of organogenic explants. However, all responding explants produced only a single adventitious
shoot irrespective of the type and concentration of the cytokinin. Adding 0.5 mg l−1 α naphthaleneacetic acid (NAA) to the medium enhanced adventitious shoot formation. In the presence of 3 mg l−1 TDZ and 0.5 mg l−1 NAA, frequency of organogenesis was 96% with a mean number of 6.1 shoots per explant. Prolonged culture or subculture on
the same medium did not promote further shoot production. However, transfer of these cultures to MS medium supplemented with
3 mg l−1 TDZ and 0.5 mg l−1 NAA and various concentrations of different polyamines (PAs), including spermine, spermidine, and putrescine, significantly
increased mean shoot number per explant. The highest frequency of shoot induction (100%) and mean shoot number per explant
(14.6) was observed on MS medium with 3 mg l−1 TDZ, 0.5 mg l−1 NAA, and 0.4 mM spermidine. Regenerated shoots were excised and subcultured on an elongation medium consisting of MS medium
with 3 mg l−1 BA. Moreover, the highest frequency of rooting (96%) and mean number of roots per shoot (3.3) was observed on MS medium with
4 mg l−1 indole-3-butyric acid (IBA) and 1.5 mg l−1 activated charcoal (AC). Almost 90% of rooted shoots were successfully acclimatized and established ex vitro. 相似文献
8.
B. K. Ghimire E. S. Seong E. J. Goh N. Y. Kim W. H. Kang E. H. Kim C. Y. Yu I. M. Chung 《Plant Cell, Tissue and Organ Culture》2010,100(2):209-217
An efficient and reproducible procedure is described for direct shoot regeneration in Drymaria cordata Willd. using leaf explants cultured on Murashige and Skoog (MS) medium supplemented with α-naphthalene acetic acid (NAA) and 6-benzyladenine. The regeneration frequency varied with the
plant growth regulator concentrations, orientation of the explants, and the carbon source and basal salts present in the regeneration
medium. The highest mean number of shoots per explant (10.65 ± 1.03) was recorded on MS plates containing 3% sucrose and 0.8%
agar supplemented with 0.1 mg/l NAA and 1.0 mg/l BAP. Shoot buds were induced in the basal parts of the leaf explants. Concentrations
of NAA exceeding 1 mg/l suppressed shoot regeneration. Explants bearing the entire lamina and petiole were much more responsive
than those having only the lamina. The plantlets that regenerated from the leaf explants were rooted successively on MS medium
alone or in combination with indole butyric acid (IBA). The highest mean number of root organogenesis, with 25.67 ± 3.68 roots
per leaf segment, was obtained in the presence of 1 mg/l IBA. Histological investigations of the regenerating shoots showed
that the shoot buds had emerged from epidermal cells without callus formation. More than 90% of the in vitro-propagated plants
survived when transferred to a greenhouse for acclimatization. Thus, this optimized regeneration system may be used for rapid
shoot proliferation and genetic transformation. 相似文献
9.
S. A. Webster S. A. Mitchell W. A. Gallimore L. A. D. Williams M. H. Ahmad 《In vitro cellular & developmental biology. Plant》2008,44(2):112-118
A procedure for producing somatic embryos enriched with dibenzyl trisulfide (DTS) using a hormone-dependent culture system
is reported for Petiveria alliacea L. (Guinea hen weed). Leaf explants were cultured on a Murashige and Skoog medium supplemented with a range of naphthaleneacetic
acid (NAA) concentrations and a fixed concentration of benzyladenine (BAP) at 11.0 μM and sucrose or glucose at 30 g l−1. Leaf explants cultured on all media types started to form callus at the cut surfaces of the discs 10–14 d after initiation.
The type of sugar used influenced average fresh weight, the propensity to form roots, as well as the embryogenic response.
The highest mean fresh weight (337.7 ± 26.18 mg) and mean root number (23.7 ± 1.69) was produced on media enriched with sucrose
and supplemented with 53.7 μM NAA and 11.0 μM BAP. An ethanol extract of rhizogenic/embryogenic callus or somatic embryos
was subjected to high-performance liquid chromatography analysis, which revealed the presence of DTS in both extracts. UV
spectral analysis and the use of standard quantitation procedures showed that the quantity of DTS in the somatic embryo extract,
at 0.16% (w/v), was approximately 30-fold higher than in rhizogenic/embryogenic callus (0.0055% w/v) of similar fresh weight. These results indicate that it is possible to biosynthesize approximately 6 mg of natural DTS from
3,808 mg of fresh somatic embryos within 10 wk from less than three leaf explants. 相似文献
10.
R. Thaniarasu T. Senthil Kumar M. V. Rao 《Physiology and Molecular Biology of Plants》2016,22(1):143-151
The present study describes the plant propagation via indirect organogenesis from in vitro derived leaf and internode explants of Plectranthus bourneae, an endemic plant to south India. Leaf and internodal explants successfully callused on Murashige and Skoog medium (MS) supplemented with different concentrations of auxins [2,4-D (2,4-dichlorophenoxyacetic acid), NAA (α-naphthalene acetic acid), IAA (indole-3 acetic acid), IBA (indole-3-butyric acid) and PIC (Picloram); 0.1–2.0 mg/l] in combination with BA (6-benzyladenine) (0.5 mg/l). Maximum callus induction (98 %) was achieved from leaf explant followed by internodal explant (89 %) at 1.0 mg/l NAA, 0.5 mg/l BA. Leaf derived callus showed better shoot regeneration (29.71 shoots) on MS medium containing 1.0 mg/l KN (kinetin), 0.7 mg/l NAA, and 50 mg/l CH (casein hydrolysate) followed by internodal callus (19.71). A maximum of 19.14 roots/shoot was observed at 1.0 mg/l IBA. The rooted plantlets were successfully hardened and transferred to greenhouse condition with 80 % survival. This system could be utilized for large-scale multiplication of P. bourneae by tissue culture. 相似文献
11.
Growth of Streptococcus zooepidemicus in a 10 l bioreactor with 50 g sucrose/l and 10 g casein hydrolysate/l gave 5–6 g hyaluronic acid/l after 24–28 h. Purification
of hyaluronic acid gave a recovery of 65% with the final material having an Mr of ∼4 × 106 Da with less than 0.1% protein. 相似文献
12.
Bimal Kumar Ghimire Chang Yeon Yu Ill-Min Chung 《Plant Cell, Tissue and Organ Culture》2012,108(3):455-464
A simple and efficient procedure was developed for in vitro propagation of Solanum aculeatissimum Jacq. using leaf and petiole explants cultured on Murashige and Skoog (MS) medium supplemented with α-naphthalene acetic
acid (NAA) and 6-benzyladenine (BA). Effects of various plant growth regulators, explant types, carbohydrates, and basal salts
on induction of adventitious shoots were also studied. Leaf explants appeared to have better regeneration capacity than petiole
explants in the tested media. The highest regeneration frequency (79.33 ± 3.60%) and shoot number (11.33 ± 2.21 shoots per
explant) were obtained in leaf explants in MS medium containing 3% sucrose and 0.8% agar, supplemented with 0.1 mg/l NAA and
2.0 mg/l BA, whereas petiole explants were more responsive to 0.1 mg/l NAA and 1.0 mg/l thiadiazuron. Developed shoots rooted
best on MS medium with 1.0 mg/l indole acetic acid (IAA), producing 18.33 ± 2.51 roots per shoot. Histological investigation
showed that the shoot buds originated mainly from epidermal cells of wounded tissues, without callus formation. The regenerated
plantlets were successfully acclimatized in a greenhouse, where over 90% developed into morphologically normal and fertile
plants. Results of flow cytometry analysis on S. aculeatissimum indicated no variation in the ploidy levels of plants regenerated via direct shoot formation and showed almost the same phenotype
as that of mother plants. This adventitious shoot regeneration method may be used for large-scale shoot propagation and genetic
engineering studies of S. aculeatissimum. 相似文献
13.
An efficient, cyclic, two-step protocol for micropropagation of medicinal tree, Crataeva nurvala has been successfully developed, which can be employed at a commercial scale. Nodal explants from 30-year-old tree when cultured
on MS medium supplemented with 2.22 μM BAP produced multiple shoots, which elongated satisfactorily on the same medium. Nodal
and leaf explants from in vitro regenerated microshoots too developed shoots, thus making the process recurrent. In 6-month
duration, owing to the recurring nature of the protocol, over 5400 shoots could be produced from a single nodal explant from
the adult tree. Addition of casein hydrolysate significantly increased the average number of shoots per explant. Maximum number
of shoots regenerated on medium supplemented with 100 mg l−1 casein hydrolysate. Shoots could be rooted on 1/2 MS supplemented with 0.11 and 0.54 μM NAA. Regenerated plantlets were acclimatized
and successfully transplanted to soil. 相似文献
14.
Yu-Ji Lian Guang-Zhe Lin Xiao-Mei Zhao Hak-Tae Lim 《In vitro cellular & developmental biology. Plant》2011,47(2):289-296
Inter-specific somatic hybrids of leaf mustard (Brassica juncea) and broccoli (Brassica oleracea) were established to introduce cytoplasmic male sterility (CMS) and Verticillium dahliae Kleb. resistance from broccoli to leaf mustard. Protoplasts isolated from hypocotyls and cotyledons of inbred lines of leaf
mustard and broccoli were fused using 40% (w/v) polyethelene glycol and then cultured in modified k8p medium supplemented with 0.2 mg/l 2,4-dichlorophenoxi-acetic acid,
0.5 mg/l 6-benzyladenine (BA), 0.1 mg/l naphthaleneacetic acid (NAA), and 0.1 mg/l kinetin (Kin), and 0.4 M mannitol as osmoticum.
At the eight- to ten-cell stage, divided cells were transferred to Kao’s basal medium supplemented with 0.3 M sucrose as carbon
source and 0.1% agarose, 2 mg/l BA, 2 mg/l zeatin (ZEA), 1 mg/l NAA, and 0.5 mg/l Kin for callus proliferation. After 35 d,
when small calli reached 2–3 mm in diameter, calli were transferred to regeneration medium containing 5 mg/l ZEA and 2 mg/l
indole-3-acetic acid. The chromosome numbers of putative somatic hybrids varied from 46 to 54. A total of ten plants showed
a 0.5-kb, CMS-specific band, while two regenerated plants showed a missing band similar to that of leaf mustard by polymerase
chain reaction amplification using Ogura CMS-specific primers. Hybrid state was confirmed by random amplified polymorphic
DNA analysis. Regenerated plants had normal petals and stamens, but only two plants produced pollen and set seed. 相似文献
15.
Micropropagation has been achieved in a promising larvicidal asteraceous taxon Spilanthes acmella L. using seedling leaf explants. The explants were reared on a variety of growth regulators, namely 2,4-dichlorophenoxyacetic
acid, 1-naphthalene acetic acid, Indole-3-butyric acid, N6-benzyladenine, and kinetin either alone or in combination on Murashige and Skoog’s (MS) medium. The best green and compact
callus was obtained on 1 μM NAA and 10 μM benzyladenine (BA) in 15 d. The callus on subculture to the same but fresh medium
after every 30 d differentiated an average of 12.90 ± 0.32 shoot buds in 50% cultures. Elongation in shoot buds occurred only
if they were transferred to NAA lacking MS+BA medium. An average number of 4.22 ± 0.83 shoots and 15 ± 0.84 shoot buds per
explant were obtained in 70.3% cultures on MS + 10 μM BA in 30 d. One hundred percent excised shoots rooted in MS(1/2) + 0.1 μM
IBA within 2 wk. The plants were gradually hardened and established in soil where they flowered and set viable seeds. The
regenerated plants were morphologically similar to the field grown plants and showed 100% larvicidal activity against malaria
and filarial vectors. 相似文献
16.
Vanilla planifolia is a tropical orchid mainly known for the aromatic flavor of its cured pods. Callus cultures were initiated from leaf and
nodal explants of V. planifolia. Leaf explants showed better callus initiation than the nodal explants with callus biomass production maximal when cultured
on Murashige and Skoog (MS) basal medium containing 4.52 mM 2,4-dichlorophenoxy acetic acid and 2.22 mM benzyladenine (BAP).
Callus transferred to MS basal medium supplemented with 13.32 μM BAP, and 13.43 μM naphthaleneacetic acid (NAA) showed superior
growth response and produced 14.0 ± 1.0 shoots with an average length of 3.6 ± 0.1 cm after 40 d. Subsequent transfer of the
proliferated shootlets to MS basal medium supplemented with 8.88 μM BAP and 8.08 μM NAA produced 12.3 ± 0.14 plantlets with
an average height of 3.6 cm ± 0.10 cm. All plantlets produced profuse rooting within 35–40 d after transfer to half-strength
MS basal medium supplemented with NAA in combination with indole-3-acetic acid. Rooted plantlets were transferred for hardening,
with 80% of the plantlets becoming successfully established in the field. Potentially, more than 100,000 plantlets could be
produced within eight subcultures from callus obtained from leaf explant through the methods described here. 相似文献
17.
Bilal Haider Abbasi Murad Khan Bin Guo Saleem Ahmed Bokhari Mir Ajab Khan 《Plant Cell, Tissue and Organ Culture》2011,105(3):337-344
The regeneration potential and antioxidative enzyme activities of economically important Brassica rapa var. turnip were evaluated. Calli were induced from leaf explants of seed-derived plantlets on Murashige and Skoog (MS) medium incorporated
with different concentrations of various plant growth regulators (PGRs). The highest leaf explant response (83%) was recorded
for 2.0 mg l−1 benzyladenine (BA) and 1.0 mg l−1 α-naphthaleneacetic acid (NAA). Subsequent subculturing of callus after 3 weeks of culture, on medium with similar compositions
of PGRs, induced shoot organogenesis. The highest shoot induction response (83%) was recorded for 5.0 mg l−1 BA after 5 weeks of transfer. However, 7.8 shoots/explant were recorded for 2.0 mg l−1 BA. The transferring of shoots to elongation medium resulted in 5.1-cm-long shoots on 10 mg l−1 of gibberellic acid (GA3). Rooted plantlets were obtained on MS medium containing different concentrations of indole butyric acid (IBA). The determination
of activities of antioxidative enzymes (superoxide dismutase [SOD], ascorbate peroxidase [APX], catalase [CAT], glutathione
peroxidase [GPX], and peroxidase [POD]) revealed involvement of these enzymes in callus formation and differentiation. All
of the activities were interlinked with each other and played significant roles in the scavenging of toxic free radicals.
This study will help in the advancement of a regeneration protocol for B. rapa var. turnip and the understanding of the functions of antioxidative enzymes in plant differentiation. 相似文献
18.
Yohana de Oliveira Fernanda Pinto André Luís Lopes da Silva Ivan Guedes Luiz Antonio Biasi Marguerite Quoirin 《In vitro cellular & developmental biology. Plant》2010,46(2):192-197
Melaleuca alternifolia is cultivated for the production of an essential oil useful in the cosmetic and pharmaceutical industries. Despite the economic
importance of this species, there is little knowledge about its in vitro propagation. The aim of this study was to establish an efficient protocol for micropropagation of M. alternifolia. With the goal of in vitro multiplication by axillary shoot proliferation, both solid and liquid MS and WPM media were tested with supplementation with
BA at 0, 0.55, 1.11, 2.22, 3.33, and 4.44 μM. The best result for shoot multiplication was obtained when either 0.55 μM BA
was added into solid MS medium or 1.11 μM BA was added into liquid MS medium, with 5.6 and 11.8 shoots per explant generated,
respectively. On solid or liquid WPM medium supplemented with 0.55 μM BA, the proliferation rates were 5.5 and 4.7, respectively.
Three auxins (NAA, IAA, and IBA) were tested at 0.53 and 2.64 μM during the rooting stage. Several sucrose concentrations
(15, 30, and 45 g L−1) were compared to a sucrose-free medium. Rooting performances on four culture media were then compared: MS, half-strength
MS (MS/2), MS + activated charcoal (AC), and MS/2 + AC. The results showed that auxin addition to culture medium is not necessary
for in vitro rooting. Rooted microcuttings from different culture media were acclimatized in a greenhouse, and the survival percentage was evaluated.
All shoots cultured in an auxin-free MS medium supplemented with sucrose (30 g L−1) produced roots, and all plants survived during acclimatization. Activated charcoal added in rooting medium reduced rooting
rates. 相似文献
19.
The genus Dierama comprises plants with a potential to be developed as ornamentals. D. erectum seeds were decontaminated and germinated on 1/10th strength Murashige and Skoog (Physiol Plant 15:473–497, 1962) (MS) media
without plant growth regulators or sucrose. In an experiment investigating the effects of 6-benzyladenine (BA), meta-Topolin (mT), kinetin (KIN) and zeatin (Z) with or without α-naphthaleneacetic acid (NAA), the highest shoot number per hypocotyl (4.20 ± 0.51)
was obtained from MS medium supplemented with 1.0 μM Z after 8 weeks. This was followed by a combination of 2.0 μM KIN and
2.0 μM NAA with 3.67 ± 0.81 shoots per explant. BA treatments produced 3.20 ± 0.22 shoots per hypocotyl explant when 2.0 μM
was combined with 1.0 μM NAA, while mT gave 3.09 ± 0.99 shoots per explant when 2.0 μM mT was combined with 2.0 μM NAA. Adventitious shoot regeneration was optimised when shoots were grown under a 16-h photoperiod
at 100 μmol m−2 s−1 on MS medium supplemented with 1.0 μM BA. This resulted in an average of 12.73 ± 1.03 shoots per hypocotyl explant. Various
concentrations of ancymidol, activated charcoal and sucrose did not promote in vitro corm formation of this species. Plants
rooted successfully after 8 weeks on MS medium supplemented with 1.0 μM indole-3-butyric acid (IBA) and had an average root
number of 2.73 ± 0.40. After 2 months of acclimatisation, plants had formed corms. The largest corms (of diameter 0.45 ± 0.03 cm)
were produced in plants pre-treated with 0.5 μM IBA. The highest plant survival percentage of 73% was also associated with
this treatment. 相似文献
20.
An efficient protocol was developed for micropropagation of an economically important timber-yielding multipurpose tree, Pterocarpus marsupium Roxb. Multiple shoots were induced from cotyledonary nodes (CNs) derived from 18-d-old axenic seedlings on Murashige and
Skoog (MS) medium supplemented with thidiazuron (TDZ) (0.1–10 μM). The highest shoot regeneration frequency (90%) and maximum
number (15.2 ± 0.20) of shoots per explant was recorded on MS medium amended with 0.4 μM TDZ. Continuous presence of TDZ inhibited
shoot elongation. In the primary medium, TDZ-initiated cultures were transferred to the secondary medium supplemented with
another cytokinin, 6-benzyladenine (BA), for shoot growth and elongation. Maximum (90%) shoot elongation with an average shoot
length of 5.4 ± 0.06 cm was observed at 5 μM BA. To further enhance the number of shoots per explant, mother tissue was repeatedly
subcultured on fresh shoot induction medium after each harvest of newly formed shoots. Thus, by adopting this strategy, an
average of 44 shoots per explant could be obtained. About 65% of in vitro regenerated shoots produced a maximum number (4.4 ± 0.2) of roots per shoot by a two-step culture procedure employing pulse
treatment and subsequent transfer of treated shoots to a low concentration of 0.2 μM indole-3-butyric acid along with phloroglucinol
(3.96 μM). The in vitro-raised plantlets were successfully acclimatized first under culture room conditions, then to greenhouse with 70% survival
rate. 相似文献