共查询到20条相似文献,搜索用时 15 毫秒
1.
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
2.
Using in vitro-grown needles of Sequoia sempervirens (D. Don.) Endl., direct shoot organogenesis was induced. The effects of three genotypes and two cytokinins, N6-benzyladenine (BA) and N-benzyl-9 (2-tetrahydropyranl) adenine (BPA), in combination with 2,4-D were investigated. Among tested cytokinins, BPA produced the highest frequency of shoot organogenesis from all three genotypes tested. Adventitious shoots were induced directly from explants without intervening callus within 5weeks following incubation. Shoots were elongated on a 1/2 Wolter and Skoog (WS) medium supplemented with activated charcoal but without growth regulators. Later, elongated shoots were transferred to a 1/4 WS medium, but without activated charcoal and free of plant growth regulators to promote continued shoot growth. These shoots rooted spontaneously. 相似文献
3.
Feng Liu Li–Li Huang Yang-Li Li Poula Reinhoud Maarten A. Jongsma Cai-Yun Wang 《Plant Cell, Tissue and Organ Culture》2011,104(1):111-117
For the first time, an in vitro regeneration protocol of Hydrangea macrophylla ‘Hyd1’ was developed. Effects of different plant growth regulators (PGRs) on shoot regeneration were investigated jointly
with selecting optimal basal media and cefotaxime concentrations. The highest frequency of shoot organogenesis (100%) and
mean number of shoots per explant (2.7) were found on Gamborg B5 basal medium supplemented with 2.25 mg/l 6-benzyladenine
(BA), 0.1 mg/l Indole-3-butyric acid (IBA), 100 mg/l cefotaxime and 30 g/l sucrose solidified by 7 g/l agar. Regenerated shoots
were rooted by culturing on perlite plus half strength liquid B5 basal medium with 0.5 mg/l NAA. Rooted plantlets were transplanted
to the greenhouse with 100% survival rate. Genetic stability of 32 plantlets (one mother plant and 31 regenerants) was assessed
by 44 ISSR markers. Out of 44 ISSR markers, ten markers produced clear, reproducible bands with a mean of 5.9 bands per marker.
The in vitro regeneration protocol is potentially useful for the genetic transformation of Hydrangea macrophylla ‘Hyd1’. 相似文献
4.
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
5.
Guohua Ma Jinfeng Lü Jaime A. Teixeira da Silva Xinhua Zhang Jietang Zhao 《Plant Cell, Tissue and Organ Culture》2011,104(2):157-162
Ochna integerrima is a medicinal and ornamental plant in Southeastern Asia. It has been listed as a rare and endangered species in China. Here
we studied the effects of plant growth regulators and their concentrations on the induction of somatic embryogenesis and shoot
organogenesis from leaf and shoot explants of O. integerrima for the first time. Cytokinins played a crucial role in somatic embryogenesis and shoot organogenesis. Among them, a higher
concentration of thidiazuron (10.0–15.0 μM TDZ) could induce both somatic embryogenesis and adventitious shoot formation whereas
low concentrations of TDZ (5.0 μM) could only induce adventitious shoots. However, 6-benzyladenine (BA at 5–15 μM) could only
induce adventitious shoots. Shoot explants induced more adventitious shoots and somatic embryos than leaf explants when cultured
on medium with the same concentration (5–15 μM) of TDZ or 15 μM BA. Medium containing 0.5 μM α-naphthaleneacetic acid and
8 μM indole-3-butyric acid and 0.1% activated charcoal could induce adventitious roots within 1 month. An efficient mass propagation
and regeneration system has been established. 相似文献
6.
Arzu Ucar Turker Esra Cansever Mutlu Arzu Birinci Yıldırım 《Acta Physiologiae Plantarum》2008,30(4):421-426
Epilobium angustifolium L. (fireweed) is a medicinal plant that has been used to treat diarrhea, mucous colitis, irritable-bowel syndrome, skin problems,
prostate problems, menstrual disorders, asthma, whooping cough, and hiccups. A highly efficient and rapid regeneration system
via multiple shoot formation was developed for fireweed. Explants (leaf, petiole, root, and stem segments) excised from sterile
seedlings were cultured on medium supplemented with different concentrations and combinations of various plant growth regulators.
Explant browning, a major problem for regeneration, was overcome by adding 100 mg/l ascorbic acid to all prepared media containing
growth regulator combinations. Root explants formed more shoots than other explants. Best shoot proliferation was obtained
from root explants cultured on media with 0.1 mg/l BA and 0.5 mg/l IAA. Regenerated shoots were transferred to rooting media
containing different concentrations of IAA, IBA, NAA or 2,4-D. Most shoots developed roots on medium with 0.5 mg/l IAA. Rooted
explants were transferred to vermiculate in Magenta containers for acclimatization and after 3 weeks they were planted in
to plastic pots containing potting soil and maintained in the plant growth room. 相似文献
7.
Guohua Ma Jaime A. Teixeira da Silva Jinfeng Lü Xinhua Zhang Jietang Zhao 《Plant Cell, Tissue and Organ Culture》2011,105(3):355-361
An efficient propagation and regeneration system via direct shoot organogenesis for an endangered species, Metabriggsia ovalifolia, was established. High activity cytokinins [6-benzyladeneine (BA) and thidiazuron (TDZ)] and low activity auxins [α-naphthaleneacetic
acid (NAA), indole-3-butyric acid (IBA) and indole-3-acetic acid (IAA)] could directly induce adventitious shoots from leaf
or petiole explants within 5 weeks. Cytokinins (TDZ or BA) combined with auxin (NAA) in the induction media induced more adventitious
shoots than when auxins or cytokinins were used alone. Adventitious shoots could be induced and also mass-propagated on media
containing 2.5–5.0 μM TDZ (or BA) and 0.25–0.5 μM NAA. Adventitious roots differentiated at the proximal end of shoots on
rooting media containing half-strength MS salts and 0.5 μM IBA, 0.5 μM NAA, 0.1% activated charcoal or no plant growth regulators.
Over 90% of plantlets survived following acclimatization and transfer to a potting mixture (1:1, sand:vermiculite) in basins. 相似文献
8.
Summary Some native species produce seeds with a low frequency of germination accompanied with a period of dormancy. These features
make it difficult to produce new phenotypes through sexual propagation. Maclura tinctoria has been considered an endangered species due to extensive use of its wood and low frequency of seed germination. The objective
of the present study is to establish an in vitro propagation system for this species. Organogenic friable callus formation from nodal segments has been obtained using woody
plant medium (WPM) supplemented with 10.74 μM 1-naphthaleneacetic acid (NAA)+4.43 μM 6-benzylaminopurine (BA). Results indicate that the highest frequency of shoot formation is observed when WPM supplemented
with 4.03 μM NAA+4.43 BA is used. For root formation, the use of WPM medium (pH adjusted to 7.0) supplemented with 23.62 μM indole-3-butyric acid (IBA) and 4.7gl−1 activated charcoal is recommended. For acelimatization, subjecting rooted plantlets to 70%, 50%, and 30% mesh screen, each
successively for a period of 7 d, has resulted in 97% plantlet survival. 相似文献
9.
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
10.
M. Sharma S. K. Rai D. K. Purshottam M. Jain D. Chakrabarty A. Awasthi K. N. Nair Ashok Kumar Sharma 《Acta Physiologiae Plantarum》2009,31(2):379-383
An in vitro process for rapid clonal propagation of Clerodendrum serratum (Linn.) Moon, a rare and threatened medicinal shrub, has been developed. Nodal stem segments having axillary bud, taken from
field-grown plant, showed bud-break within 15 days of culture on modified Murashige and Skoog (MS) (Physiol Plant 15:473–497,
1962) medium supplemented with 0.25 mg/l each of 6-benzylaminopurine and indole-3-acetic acid along with 15 mg/l adenine sulphate
(AdS). Regenerated shoots could be further multiplied on the same agarified morphogenetic medium in presence of 0.5 mg/l 2-chloroethyltrimethyl
ammonium chloride with increased concentration of AdS, i.e., 30 mg/l. A group of five shoots used as inoculum produced on
an average 4.98 new shoots per original shoot after 4 weeks of subculture. Shoots excised from cultures of proliferating shoots
were rooted in half-strength MS medium having 1 mg/l indole-3-propionic acid. In vitro rooted shoots—plantlets—grew luxuriantly
under field conditions and came to flowering after 10 months of transplantation. The genetic fidelity of in vitro-raised field-grown
plants and their mother plant was ascertained by random amplified polymorphic DNA markers. The protocol developed holds good
for in vitro cloning of C. serratum. 相似文献
11.
A rapid and efficient procedure is outlined for in vitro clonal propagation of an elite cultivar of jewel orchid (Anoectochilus formosanus). Multiple shoot proliferation was induced in shoot tip explants on Hyponex (H3) media supplemented with 1 mg dm–3 benzyladenine or 1 – 2 mg dm–3 thidiazuron (TDZ). Addition of activated charcoal (1 g dm–3) to the TDZ containing medium promoted multiple shoot formation (11.1 shoots per explant). However, the regenerated shoots had slow growth rate and failed to elongate. This problem was overcome by transferring the shoot clumps to a hormone free H3 medium supplemented with 2 % sucrose and 0.5 g dm–3 activated charcoal. Rooting was induced in 100 % of the regenerated shoots in the same media. The plantlets were acclimatized and established in greenhouse. 相似文献
12.
Songul Gurel Mehmet Cengiz Baloglu Ekrem Gurel Huseyin Avni Oktem Meral Yucel 《Plant Cell, Tissue and Organ Culture》2011,106(2):261-268
The effects of a two-stage pretreatment of seedlings on the subsequent shoot regeneration capacity were investigated. Pretreated
seedlings were obtained by germinating seeds on three different germination media and then further culturing on six different growth media. Lamina and petiole explants of two sugar beet (Beta
vulgaris L.) breeding lines were then excised from the pretreated seedlings and cultured on five different shoot regeneration media. In both breeding lines, petiole explants produced significantly more shoots than lamina explants with higher frequencies
of organogenic capacities; petiole explants of the lines M1195 and ELK345 produced a mean of 2.1 and 2.7 shoots per explant
while their lamina explants produced 1.5 and 2.2 shoots per explant, respectively. A genotypic variation was evident as the
line ELK345 was more productive for shoot development from both types of explants. In overall comparisons of different germination, growth and regeneration media, germination medium was most effective when supplemented with 0.5 mg/l 6-benzyladenine (BA) while both growth and regeneration
media were most productive when contained a combination of 0.25 mg/l BA and 0.10 mg/l indole-3-butyric acid (IBA). Of all
the treatments tested, the highest mean number of shoots per explant (8.3 shoots) and frequency of organogenic explants (75.6%)
were obtained on regeneration medium supplemented with 0.25 mg/l BA and 0.10 mg/l IBA when petiole explants of the line ELK345
were excised from the seedlings that had been germinated on medium containing 0.5 mg/l BA followed by further growth on medium
containing 0.25 mg/l BA and 0.10 mg/l IBA. 相似文献
13.
Sandra Gonçalves Laura Fernandes Anabela Romano 《Plant Cell, Tissue and Organ Culture》2010,101(3):359-363
A novel protocol suitable for the micropropagation of the endangered species Tuberaria major using seedlings as explants is reported. Using this protocol, we studied the effects of explant type (apical shoots and nodal
segments) and cytokinins [6-benzyladenine (BA), kinetin, and zeatin (ZEA)] on shoot proliferation. Explant type significantly
influenced the proliferation frequency and mean number of shoots, with nodal segments showing a higher proliferation capacity.
The mean number of shoots was significantly higher when the explants were cultured in half-strength (1/2) MS medium supplemented
with 0.2 mg l−1 BA (6.83 ± 0.77 shoots) or ZEA (6.55 ± 0.71 shoots). The shoots showed a great rooting capacity that was significantly influenced
by the concentration of MS macronutrients but not by the concentration of auxins. The highest rooting frequencies (97–100%)
were obtained in 1/2 MS medium with or without plant growth regulators. The plants obtained were easily acclimatized to ex
vitro conditions, with 97% surviving after 6 weeks. The micropropagated plants were successfully reintroduced into their natural
habitat and exhibited normal development. In conclusion, our culture protocol, with efficient seed germination, subsequent
multiplication of nodal explants using ZEA at 0.2 mg l−1, and successful ex vitro establishment of well-rooted plantlets on 1/2 MS medium, provides a simple and reliable methodology
for the large-scale propagation of T. major, thereby contributing to germplasm preservation of this endangered species. 相似文献
14.
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
15.
Semecarpus anacardium (Anacardiaceae), a deciduous forest tree, is a potent source of medicinal compounds. Poor seed viability of this species
limits the conventional propagation practice. Proliferation of shoots from axillary meristem was achieved in semisolid WPM
medium supplemented with BAP 4.44 μM and KN 4.64 μM. Factors including culture vessels, gelling agents and antioxidants were
identified and optimized for proliferation and growth of shoots in vitro. Cotton-plugged culture vessels were more favorable.
Phytagel 0.2% as gelling agent and activated charcoal 0.2% as antioxidant were superior to other agents and antioxidants tested.
All the shoots rooted in half-strength WPM liquid medium with IBA 2.46 μM. Rooted shoots survived (91%) in the soil–sand 1:1
mixture. Ex vitro rooting of shoots and hardening of plants were achieved in 80% of the explants in the soil–sand mixture.
Hardened plants were maintained in a greenhouse. This is the first report on in vitro regeneration of Semecarpus anacardium. 相似文献
16.
An efficient in vitro plant regeneration protocol for Swertia chirata Buch.-Ham. ex Wall (Gentianaceae), a critically endangered Himalayan medicinal herb, was developed using shoot tip explants
derived from in vitro grown seedlings. Media with 2% sucrose and various types of hormones markedly influenced in vitro propagation
of S. chirata. An in vitro shootlet production system using Murashige and Skoog (MS) medium with various hormones such as BAP, KN and TDZ
was established. BAP at 1.0 mg/l and KN, 0.1 mg/l induced highest number of multiple shoots (42.16 ± 1.05) per explant. Micro-proliferated
shoots were transferred to elongation medium amended with GA3 (0.1 mg/l) and hormone free basal medium, after which they were transferred to rooting medium. The highest frequency of rooting
(22.48 ± 1.08) was obtained in half-strength MS medium supplemented with NAA, 0.1 mg/l after testing with different auxins
at various concentrations within 4 weeks of transfer to the rooting medium. Hardening was successfully attained under controlled
conditions inside the plant tissue culture room. This method could effectively be applied for the conservation and clonal
propagation to meet the pharmaceutical demands. 相似文献
17.
A rapid and efficient in vitro plant regeneration method was developed for Matteuccia struthiopteris (L.) Todaro (Ostrich fern). Side shoots, originating in meristems of sectioned rhizomes, were used as explant material. A
very high rate of meristem multiplication was achieved by culturing the explants in half-strength MS liquid medium supplemented
with 2.0 mg/l N-(4-Pyridyl)-N′-phenylurea (4-PU) and 0.5 mg/l thidiazuron (TDZ). Multiplication of the shoot primordia was
faster in suspension culture than on solid medium. Rhizogenesis and growth of regenerants were best achieved on hormone-free
one-quarter-strength MS solid medium amended with 0.4% agar and 1.0% activated charcoal. Regenerated plantlets continued to
grow after transfer to soil in a phytotron.
Received: 19 March 1998 / Revision received: 17 July 1998 / Accepted: 3 August 1998 相似文献
18.
Zhen Xu Yeong-Cheol Um Chun-Hwan Kim Gang Lu De-Ping Guo Hai-Lin Liu Amadou A. Bah Aining Mao 《Acta Physiologiae Plantarum》2008,30(4):521-528
In vitro shoot proliferation from stem disc of Allium chinense, a vegetatively propagated plant, was investigated in this experiment. In the present study, shoots were formed directly
on stem discs on a medium containing 1 mg/l N6-benzyladenine (BA) and 0.5 mg/lα-naphthaleneacetic acid (NAA). These shoots were further cultured on MS media supplemented
with various levels of BA in combination with NAA, and new shoot clusters developed easily from the explants cultured despite
considerable differences in the induction of shoot clusters with different levels of BA and NAA. The most productive combination
of growth regulators proved to be 1.0 mg/l BA and 1.0 mg/l NAA, in which about 17 shoots were produced per cluster in 8 weeks
culture. Most of the formed shoots were rooted 15 days after being cultured on MS media supplemented with 0.1–1.0 mg/l NAA.
The survival rate of the plantlets under ex vitro conditions was 95% in pots filled with a peat: sand (2:1 v/v) mixture after
two weeks. In vitro bulblet formation were strongly promoted by the high temperature of 30°C compared to that at 25, 20 and
15°C, and 12% (w/v) sucrose appeared to be optimal for bulblet development. Results from this study demonstrated that A. chinense could be in vitro propagated by using stem discs and in vitro bulblet formation could be achieved. 相似文献
19.
Seied Ali Hosseini Tafreshi Mansour Shariati Mohammad Reza Mofid Mojtaba Khayam Nekui 《In vitro cellular & developmental biology. Plant》2011,47(5):561-568
A highly promising procedure to obtain seedlings of Taxus baccata L. has been developed, which involves a combination of in vitro embryo culture and growth under hydroponic conditions. Embryos isolated from freshly collected seeds were 100% sterile, even
though the seeds were not treated with acid or soaked in water prior to culturing. The embryo germination level of non-leached
seeds was slightly lower (85%) than those leached in running water for 7 d (100%). The leached embryos germinated with extended
roots while the non-leached embryos had abnormal shapes. The embryos cultured on media supplemented with an absorbent (PVP
or activated charcoal) had extended roots and shoots and were a larger size without any browning, as compared to those grown
without the supplement; activated charcoal gave better results. There were no significant differences in germination rates
of T. baccata embryos between the media with differing strengths of macronutrients; however, for further development of the shoot, it was
necessary to sub-culture the seedlings in MS in the light. To obtain seedlings with longer roots, they had to be maintained
in one-half strength MS in darkness. Approximately 90% of the plants survived when grown hydroponically for 2 mo. The surviving
plants showed well-extended roots and were a good starting material for genomic, proteomic, and conservational studies as
well as Taxol permeabilization investigations. 相似文献
20.
Immature embryos of Quercus acutissima were collected weekly beginning 5 weeks post-fertilization and cultured on modified MS(Murashige and Skoog) medium containing 1,000 mg/l glutamine and 5 mM proline with different combinations of IBA(0.5–10.0 mg/l) and BA(0 or 1.0 mg/l) in light. The highest percentage of embryogenic cultures occurred on the medium containing 0.5 mg/l IBA or 1.0 mg/l BA and 0.5 mg/l IBA. Four weeks after initiation, the embryogenic cultures were transferred to MS medium without plant growth regulators and cultured for 4 weeks. The somatic embryos were then transferred to germination medium. The best germination results were achieved from WPM(Woody Plant Medium) containing 0.1 mg/l BA. Plantlets from somatic embryos were incubated on WPM supplemented with 0.2 mg/l BA for 4 weeks and plantlets with well developed shoots and roots were transplanted to perlite and peat moss(11, v/v) mixtures and placed in a culture room. After being hardened off for 8 weeks, they were transferred outdoors where they grew.Abbreviation BA
N6-benzyladenine
- IBA
indole-3-butyric acid
- GA3
gibberellic acid
- ABA
abscisic acid
- MS
Murashige & Skoog Medium
- WPM
Woody Plant medium 相似文献