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A. Śliwińska O. Olszowska M. Furmanowa A. Nosov 《In vitro cellular & developmental biology. Plant》2008,44(2):69-77
Efficient plant regeneration through somatic embryogenesis was achieved in Polyscias filicifolia. Embryogenic calluses were induced on Murashige and Skoog (MS) basal medium supplemented with 0.5 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg l−1 benzylaminopurine (BAP; type I callus) and on MS medium with 2.0 mg l−1 2,4-D and 0.01 mg l−1 kinetin (type II callus) from leaf explants of a 2-yr-old plant. Primary somatic embryos (PSEs) developed after four passages
of suspension culture established from embryogenic callus when cultured in liquid half-strength MS medium (1/2 MS) without
growth regulators. PSEs in the cotyledonary stage were multiplied by adventitious embryogenesis. Single secondary somatic
embryos (SSEs) or their clusters developed at the base of PSE hypocotyls and regenerated into plantlets in a one-step process
on plant growth regulator-free 1/2 MS medium. Low sucrose concentration of 15 g l−1 promoted development of normal SSEs. All SSEs regenerated into single, well-rooted plantlets on a Nitsch and Nitsch medium
supplemented with 0.5 mg l−1 kinetin, 0.1 mg l−1 indole-3-butyric acid, and 10 mg l−1 adenine sulfate. Subsequent two subculture cycles on the same medium were necessary to obtain plantlets sufficiency developed
to allow successful transfer to the soil. Rooted plantlets were established in a peat mixture with 90% survival, with the
plants showing normal morphological characteristics. 相似文献
3.
Plant regeneration through indirect somatic embryogenesis has been established on Holostemma ada-kodien Schult. Type of auxin significantly influenced somatic embryogenesis. Friable callus, developed from leaf, internode and root explants on Murashige and Skoog (MS) medium supplemented with 2,4-D (1.0 mg l–1), was most effective for the induction of somatic embryos. Subculture of the friable callus developed on 2,4-D (1.0 mg l–1) onto solid or liquid 1/2 MS medium with 0.1 or 0.5 mg l 2,4-D turned the callus embryogenic. Suspension cultures were superior to static cultures (solid medium) for the induction of somatic embryos. Transfer of embryogenic callus to liquid 1/2 or 1/4 MS medium with lower levels of 2,4-D (0.05–0.1 mg l–1) induced the highest number of somatic embryos. An average of 40 embryos were obtained from 10 mg callus. Fifty per cent embryos exhibited maturation and conversion upon transfer to 1/10 MS basal solid medium. Plantlets were established in field conditions and 90 per cent survived. 相似文献
4.
Yongxue Yang Guofeng Liu Manzhu Bao 《In vitro cellular & developmental biology. Plant》2006,42(6):520-524
Summary A protocol of somatic embryogenesis and plant regeneration from petiole segments of Parthenocissus tricuspidata Planch. has been developed. Embryogenic tissue was induced on B5 (Gamborg) basal medium supplemented with 2.25–9.0 μM 2,4-dichlorophenoxyacetic acid, 500 mg l−1 casein hydrolysate (CH), and 0.1 gl−1 activated charcoal. Somatic embryos were induced on B5 medium containing various concentrations of benzyladenine (BA) (4.44,
6.66, and 8.88 μM) and α-naphthaleneacetic acid (NAA) (0, 0.54, and 1.61 μM) plus 500 mg l−1 CH. Ninety percent of normal somatic embryos were converted into plantlets directly on Murashige and Skoog (MS) medium free
of plant growth regulators. Shoots could be induced from abnormal somatic embryos on MS medium containing 4.44 μM BA, 0.05 μM NAA, and 500 mg l−1 CH. Genotypic differences were found in the process of somatic embryogenesis and plant regeneration. Histological analysis
confirmed the process of somatic embryogenesis. Regenerated plantlets with well-developed roots were successfully acclimatized
in greenhouse and all plants showed normal morphological characteristics. 相似文献
5.
A system for rapid plant regeneration through somatic embryogenesis from shoot tip explants of sorghum [Sorghum bicolor (L.) Moench] is described. Somatic embryogenesis was observed after incubation of explants in dark for 6–7 weeks through
a friable embryogenic callus phase. Linsmaier and Skoog medium supplemented with 2,4-dichlorophenoxyacetic acid (2 mg l−1) and kinetin (0.1 mg l −1) was used for induction of friable embryogenic calli and somatic embryos. Germination of somatic embryos was achieved about
5 weeks after transfer onto Murashige and Skoog (MS) medium supplemented with 6-benzylaminopurine (2 mg l−1) and indole-3-acetic acid (0.5 mg l −1) under light. Seeds from in vitro-regenerated plants produced a normal crop in a field trial, and were comparable to the crop grown with the seeds of the mother
plant used to initiate tissue culture. The simplicity of the protocol and possible advantages of the system for transformation
over other protocols using different explants are discussed.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
6.
Direct somatic embryogenesis of Frittilaria meleagris L. was induced using leaf base explants excised from in vitro grown shoots. Somatic embryos occurred at the basal part of leaf explants 4 weeks after culture on a Murashige and Skoog
(MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) or kinetin (KIN). The highest
number of somatic embryos (SEs) were formed (9.74) from leaf explant on MS medium supplemented with 0.1 mg dm−3 2,4-D after 4 weeks of culture initiation. An initial exposure to a low concentration of KIN in the medium also enhanced
SEs induction. Our observations by light and scanning electron microscopy revealed that SEs originate directly from the epidermal
and subepidermal layers of leaf explant. The developmental stages of somatic embryogenesis from the first unequal cell division
through the meristematic clusters, multi-cellular globular somatic embryos to the fully formed cotyledonary embryos were determined.
After 4 weeks on MS medium without plant growth regulators, SEs developed into bulblets. 相似文献
7.
Plant regeneration was achieved through direct and indirect somatic embryogenesis in Eucalyptus camaldulensis. Callus was induced from mature zygotic embryos and from cotyledon explants collected from 10, 15, 25, and 30-day-old seedlings
cultured on Murashige and Skoog (MS) basal medium supplemented with different concentrations of naphthaleneacetic acid (NAA).
Maximum callus induction from mature zygotic embryos was obtained on MS basal medium containing 1 mg l−1 NAA. The frequency of callus development varied based on the age of the cotyledon explants 10-day-old explants giving highest
percentage on MS basal medium supplemented with 1 mg l−1 NAA. Callus obtained from mature zygotic embryos gave highest frequency of somatic embryogenesis on MS basal medium containing
0.5 mg l−1 benzyladenine (BA) and 0.1 mg l−1 NAA. Separate age wise culture of the calli, obtained from cotyledons of different ages cultured separately, revealed high
somatic embryogenic potential on callus from 10-day-old cotyledons. Direct somatic embryogenesis too was obtained from hypocotyl
explants without an intervening callus phase on MS basal medium containing 0.5 mg l−1 BA. The effects of abscisic acid (ABA), sucrose, and different strengths of MS medium on somatic embryo maturation and germination
were also investigated. Number of mature somatic embryos increased with lower concentrations (0–1 mg l−1) of ABA while no significant differences were observed at higher concentrations (2–5 mg l−1) of ABA. Compared to basal medium containing lower concentrations of sucrose (1%), the MS medium supplemented with higher
levels of sucrose (4%) showed significantly lower frequency of mature somatic embryos. Basal medium without any dilution gave
the highest number of immature embryos. However, the number of mature embryos was high at higher medium dilutions. 相似文献
8.
Several plant growth regulators BA, TDZ, 2,4- and Kn were tested alone or in combination for their capacity to induce indirect somatic embryogenesis from leaf and internode explants of Paulownia elongata. Calli were produced when leaf explants were cultured on Murashige and Skoog (MS) medium containing 3 % sucrose, 0.4 % phytagel, 4 mg l-1 TDZ and 0.1 mg l-1 Kn after 3 weeks and the initiation rate was 54.1%. After subculturing on the same medium, embryos at various developmental stages (globular, heart and torpedo shaped) were transferred for maturation onto MS medium supplemented with 3 % sucrose, 0.4 % phytagel, 0.1 mg l-1 TDZ, 1 mg l-1 Kn and 2 mM glutamine. An average of 50.7 somatic embryos were obtained from 100 mg of embryogenic callus after 4 weeks at high frequency (64.7 %). Afterward, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination (80 %) and development into plantlets. Plantlets were transferred to pots with a mixture of peat and perlite in a 3:1 ratio and showed a survival rate of 70–80 %. Plantlets regenerated by this procedure were morphologically identical to the donor material and developed normally in the greenhouse. 相似文献
9.
Archana Giri Paramvir Singh Ahuja P. V. Ajay Kumar 《Plant Cell, Tissue and Organ Culture》1993,32(2):213-218
Plants were obtained via somatic embryogenesis in callus derived from in vitro raised leaf and petiole explants of Aconitum heterophyllum Wall. Callus was induced on a Murashige-Skoog medium supplemented with either 2,4-dichlorophenoxy acetic acid (2,4-d 1 mg l-1) and kinetin (KN 0.5 mg l-1) with coconut water (CW 10% v/v) or naphthalene acetic acid (NAA 5 mg l-1) and benzylaminopurine (BAP 1 mg l-1). Somatic embryos appeared after 2–3 months or 2 subculture passages when 2,4-d or NAA induced source of the callus was transferred to a MS medium containing BAP (1 mg l-1) and NAA (0.1 mg l-1). For successful plantlet formation, the somatic embryos were transferred to a medium containing 1/4 strength MS nutrient with indole-3-butyric acid (IBA 1 mg l-1). Alternatively, the somatic embryos were dipped in a concentrated solution of IBA for 5 min and placed on a hormone free medium. Complete plantlets were formed after 4 weeks and were transferred successfully to soil.CIMAP Publication No. 1020. 相似文献
10.
A three-stage procedure for embryogenesis in Trachyspermum ammi was developed from cotyledon and cotyledonary node explants cultured in Murashige and Skoog (MS) liquid medium supplemented
with 0.2 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Globular somatic embryos without intervening callus phase developed in 4 wk. The
development of embryos to heart and torpedo stages required second-stage subculture of the explants (along with developing
embryos) in liquid medium with lower concentrations of 2,4-D. Further development of embryos required a third-stage subculture
in hormone-free liquid medium supplemented with 100 mg l−1 casein hydrolysate. Regeneration of complete plantlets occurred after the fully developed somatic embryos were transferred
to solidified half-strength MS medium supplemented with 1 mg l−1 gibberellic acid. 相似文献
11.
Myung Jin Oh Hye Ryun Na Hong-Keun Choi Jang Ryol Liu Suk Weon Kim 《Plant biotechnology reports》2008,2(1):87-92
An improved protocol for high frequency plant regeneration via somatic embryogenesis from zygotic embryo-derived cell suspension
cultures of watershield (Brasenia schreberi) was developed. Zygotic embryos formed pale-yellow globular structures and white friable callus at a frequency of 80% when
cultured on half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. However, the frequency of formation of pale-yellow globular structures and white friable callus decreased slightly
with increasing concentrations of 2,4-D up to 3 mg l−1, where the frequency reached ~50% of the control. Cell suspension cultures from zygotic embryo-derived white friable callus
were established using half-strength MS medium supplemented with 0.3 mg l−1 2,4-D. Upon plating of cell aggregates on half-strength MS basal medium, approximately 8.3% gave rise to somatic embryos
and developed into plantlets. However, the frequency of plantlet development from cell aggregates was sharply increased (by
up to 55%) when activated charcoal and zeatin were applied. Regenerated plantlets were successfully transplanted to potting
soil and grown to normal plants in a growth chamber. The distinctive feature of this study is the establishment of a high
frequency plant regeneration system via somatic embryogenesis from zygotic embryo-derived cell suspension cultures of watershield,
which has not been previously reported. The protocol for plant regeneration of watershield through somatic embryogenesis could
be useful for the mass propagation and transformation of selected elite lines. 相似文献
12.
Plant regeneration through somatic embryogenesis from mature leaf explants of Eryngium foetidum,a condiment 总被引:1,自引:0,他引:1
Ignacimuthu S. Arockiasamy S. Antonysamy M. Ravichandran P. 《Plant Cell, Tissue and Organ Culture》1999,56(2):131-137
Eryngium foetidum L. is an important plant cultivated as a leafy vegetable and for its essential oil, which are of high economic
value in international trade market. Plants were regenerated through somatic embryogenesis from mature leaf explants of field
grown plants. Leaf explants produced dark brown, compact callus on Linsmaier and Skoog (LS) medium with the combination of
1.0 mg l-1 2,4-dichlorophenoxy acetic acid (2,4-D) and 1.0 mg l-1 benzylaminopurine (BAP). Somatic embryos were induced from embryo-forming callus cultures on Murashige and Skoog (MS) medium
supplemented with 0.1 mg l-1 2,4-D, 2.0 mg l-1 BAP and 1.0 mg l-1 gibberellic acid (GA3). Subsequently, conversion of these somatic embryos into plantlets occurred on MS medium supplemented with 1.0 mg l-1 GA3 and/or 0.1 mg l-1 BAP. The regenerated shoots were rooted and elongated on MS medium supplemented with 0.1 mg l-1 IAA and 1.0 mg l-1 GA3. These plantlets were hardened and transferred to the soil.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
13.
Yu-Shi Luan Juan Zhang Xiao-Rong Gao Li-Jia An 《Plant Cell, Tissue and Organ Culture》2007,88(1):77-81
Salt tolerant cultivars of sweet potato (Ipomoea batatas L.) can be obtained from induced mutation. The objective of the present study was to induce mutation for salt tolerance using
ethylmethanesulphonate (EMS) in calli of sweet potato, followed by cell line selection and subsequent plant regeneration.
Calli initiated from leaf explants were treated with 0.5% EMS for 0, 1, 1.5, 2, 2.5 and 3 h, followed by rinsing with sterile
distilled water for four times. Preliminary experiments showed that 200 mM NaCl could be used as selection pressure. Salt
tolerant calli were sub-cultured on medium supplemented with 200 mM NaCl for selection of mutant cell lines and this process
repeated 5 times (20 days each). The selected calli were transferred onto somatic embryo formation medium, which was Murashige
and Skoog (MS) medium supplemented with 4 mg l−1 abscisic acid (ABA), 10 mg l−1 gibberellic acid (GA). After 15 days, somatic embryos were transferred onto MS medium supplemented with 0.05 mg l−1 ABA, 0.2 mg l−1 zeatin (ZT) for regeneration. Plants designated as ML1, ML2 and ML3 were regenerated from the somatic embryos formed by calli
treated with 0.5% EMS for 2 and 2.5 h. After propagation, salt tolerance of these mutants was investigated. Data suggested
the mutants were more salt tolerant than control plants. 相似文献
14.
Summary A method for the production of somatic embryos and subsequent plant regeneration for Anthurium andraeanum Linden ex André (Monocotyledonae) hybrids is described. Whole leaf blade explants, derived from plantlets grown in vitro, formed translucent embryogénic calli at their basal ends within one month of culture in the dark. Secondary somatic embryos formed frequently and without an intervening callus on surfaces of primary embryos. Embryogenesis was induced with three genotypes using a modified half-strength Murashige and Skoog (MS) medium supplemented with 1.0 to 4.0 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) and 0.33 to 1.0 mg l–1 kinetin. A combination of 2% sucrose with 1% glucose in the medium favored embryogenesis over 3% sucrose alone. Whole leaf blades on medium solidified with 0.18% Gelrite produced more somatic embryos than leaves on medium with 0.7% Bacto-agar. Within two to three months after culture initiation, embryos were transferred to modified MS medium containing 0.2 mg l–1 6-benzyladenine (BA) and 2% sucrose and placed in the light for conversion into plantlets. Rooted plantlets were recovered and transferred into pots with tree fern fiber medium and grown in the greenhouse.Abbreviations MS
Murashige and Skoog (1962)
- 2,4-D
2,4 dichlorophenoxyacetic acid
- BA
6-benzyladenine 相似文献
15.
Michael E. Compton Jennifer M. Koch 《In vitro cellular & developmental biology. Plant》2001,37(2):259-261
Summary The influence of PPMTM on somatic embryogenesis in melon, adventitious shoot organogenesis in petunia, and androgenesis in tobacco was studied by
culturing explants in regeneration media supplemented with 0, 2, 5 or 10 ml l−1 PPM for 8–12 wk. The percentage of melon cotyledon explants that produced callus and somatic embryos and the number of embryos
per explant were reduced when incubated in embryo initiation and embryo development media containing more than 5 ml l−1 PPM. Less PPM was required to inhibit petunia shoot organogenesis. The number of shoots and number of buds per Petri dish
were reduced 3–6.9-fold when leaf explants were incubated in shoot regeneration medium containing more than 2 ml l−1 PPM. In contrast, the addition of up to 10 ml l−1 PPM to tobacco anther culture medium had no effect on androgenesis. Our results suggest that the influence of PPM on plant
regeneration depends on the plant species. We recommend that experimenters examine a range of PPM concentrations when using
it for the first time on an untested plant species. 相似文献
16.
P. Giridhar Vinod Kumar G. A. Ravishankar 《In vitro cellular & developmental biology. Plant》2004,40(6):567-571
Summary A novel protocol has been developed for inducing somatic embryogenesis from leaf cultures of Decalepis hamiltonii. Callus was obtained from leaf sections in Murashige and Skoog (MS) medium supplemented with α-naphthaleneacetic acid (NAA)+N6-benzyladenine (BA) or 2,4-dichlorophenoxyacetic acid (2,4-D)+BA. Nodular embryogenic callus developed from the cut end of
explants on media containing 2,4-D and BA, whereas compact callus developed on media containing NAA and BA. Upon subsequent
transfer of explants with primary callus onto MS media containing zeatin and/or gibberellic acid (GA3) and BA, treatment with zeatin (13.68μM) and BA (10.65 μM) resulted in the induction of the highest number of somatic embryos directly from nodular tissue. The maturation of embryos
took place along with the induction on the same medium. Embryogenic calluses with somatic embryos were subcultured onto MS
basal medium supplemented with 4.56μM zeatin+10.65 μM BA. After 4wk, more extensive differentiation of somatic embryos was observed. The mature embryos developed into complete
plantlets on growth regulator-free MS medium. A distinct feature of this study is the induction of somatic embryogenesis from
leaf explants of Decalepis hamiltonii, which has not been reported previously. By using this protocol, complete plantlets could be regenerated through indirect
somatic embryogenesis or organogenesis from leaf explants in 12–16 wk. 相似文献
17.
Zhong Hu Yi-Rui Wu Wei Li Huan-Huan Gao 《In vitro cellular & developmental biology. Plant》2006,42(5):461-466
Summary Using the system for genetic transformation and transgenic plant regeneration via somatic embryogenesis (SE) of Lycium barbarum established in this laboratory, this study reports the optimization of the factors affecting the efficiency of transformation,
including pre-culture period, leaf explant source, use of acetosyringone, strains and density of Agrobacterium, and temperature of co-cultivation. The optimized transformation protocol for L. barbarum included preculture of leaf explants from 3-wk-old seedlings for 3 d on the medium for callus induction followed by inoculation
with Agrobacterium strain EHA101 (pIG121 Hm), co-cultivation for 3d at 24°C, and transfer to the selection regeneration medium with 50
mg l−1 kanamycin (Kan). Using this protocol, 65% L. barbarum explants gave rise to Kan-resistant and GUS-positive calli. In addition, the expression of introduced transgene (npt II) in clonal progeny was verified by formation of calli and somatic embryos from leaf segments of nine transgenic plants
grown on the Kan-containing medium. All explants formed calli at 50 mg l−1 Kan and seven out of nine transgenic plants were found to possess callus-forming capacity even at 100 mg l−1 Kan. These calli also possessed higher SE potential on SE medium supplemented with 25 mg l−1 Kan. 相似文献
18.
Several plant growth regulators BA, TDZ, 2,4- and Kn were tested alone or in combination for their capacity to induce indirect somatic embryogenesis from leaf and internode explants of Paulownia elongata. Calli were produced when leaf explants were cultured on Murashige and Skoog (MS) medium containing 3 % sucrose, 0.4 % phytagel, 4 mg l-1 TDZ and 0.1 mg l-1 Kn after 3 weeks and the initiation rate was 54.1%. After subculturing on the same medium, embryos at various developmental stages (globular, heart and torpedo shaped) were transferred for maturation onto MS medium supplemented with 3 % sucrose, 0.4 % phytagel, 0.1 mg l-1 TDZ, 1 mg l-1 Kn and 2 mM glutamine. An average of 50.7 somatic embryos were obtained from 100 mg of embryogenic callus after 4 weeks at high frequency (64.7 %). Afterward, mature somatic embryos were isolated and cultured on hormone-free MS medium for germination (80 %) and development into plantlets. Plantlets were transferred to pots with a mixture of peat and perlite in a 3:1 ratio and showed a survival rate of 70–80 %. Plantlets regenerated by this procedure were morphologically identical to the donor material and developed normally in the greenhouse. 相似文献
19.
Direct somatic embryogenesis and synthetic seed production from<Emphasis Type="Italic"> Paulownia elongata</Emphasis> 总被引:8,自引:0,他引:8
We have developed a reproducible system for efficient direct somatic embryogenesis from leaf and internodal explants of Paulownia elongata. The somatic embryos obtained were subsequently encapsulated as single embryos to produce synthetic seeds. Several plant growth regulators [6-benzylaminopurine, indole-3-acetic acid, -naphthaleneacetic acid, kinetin and thidiazuron (TDZ)] alone or in combination were tested for their capacity to induce somatic embryogenesis. The highest induction frequencies of somatic embryos were obtained on Murashige and Skoog (MS) medium supplemented with 3% sucrose, 0.6% Phytagel, 500 mg l-1 casein hydrolysate and 10 mg l-1 TDZ (medium MS10). Somatic embryos were induced from leaf (69.8%) and internode (58.5%) explants on MS10 medium after 7 days. Subsequent withdrawal of TDZ from the induction medium resulted in the maturation and growth of the embryos into plantlets on MS basal media. The maturation frequency of somatic embryos from leaf and internodal explants was 50.8% and 45.8%, respectively. Subculturing of mature embryos led to their germination on the same medium with a germination frequency of 50.1% and 29.8% from leaf and internode explants, respectively. Somatic embryos obtained directly on leaf explants were used for encapsulation in liquid MS medium containing different concentrations of sodium alginate with a 30-min exposure to 50 mM CaCl2. A 3% sodium alginate concentration provided a uniform encapsulation of the embryos with survival and germination frequencies of 73.7% and 53.3%, respectively. Storage at 4°C for 30 days or 60 days significantly reduced the survival and complete germination frequencies of both encapsulated and non-encapsulated embryos relative to those of non-stored somatic embryos. However, the survival and germination rates of encapsulated embryos increased following storage at 4°C. After 30 days or 60 days of storage, the survival rates of encapsulated embryos were 67.8% and 53.5% and the germination frequencies were 43.2% and 32.4%, respectively. These systems could be useful for the rapid clonal propagation and dissemination of synthetic seed material of Paulownia elongata.Abbreviations BAP 6-Benzylaminopurine - IAA Indole-3-acetic acid - NAA -Naphthaleneacetic acid - TDZ ThidiazuronCommunicated by H. Lörz 相似文献
20.
Root explants excised from carnation plants maintained in vitro formed off-white, friable calluses after three weeks of culture
on Murashige and Skoog (MS) medium supplemented with 1 mg l−1 thidiazuron (TDZ) and 1 mg l−1 α-naphthalaneacetic acid (NAA). These calluses were subsequently transferred to MS basal medium where, after an additional
four weeks of culture, approximately 50% of the calluses formed somatic embryos. However, calluses formed on root explants
that had been cultured on MS medium supplemented with 2,4-dichlorophenoxyacetic acid did not produce somatic embryos upon
transfer to MS basal medium. Somatic embryos developed into plantlets and subsequently were grown to maturity. These results
indicate that root explants have a high competence for somatic embryogenesis in carnation.
J. Seo and S.W. Kim contributed equally to this work. 相似文献