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1.
Summary A somatic embryogenesis protocol was developed for Eschscholzia californica Chan. (California poppy) using embryogenic cell suspensions and optimized media conditions. Rapidly-growing, finely-dispersed
embryogenic cell suspension cultures were established from embryogenic callus and maintained in B5 liquid media supplemented
with 0.5 mg 1−1 (2.26 μM) 2,4-dichlorophenoxyacetic acid. Culture conditions were optimized by investigating the effect of basal media composition,
gyratory shaker speed, various carbon sources, different cytokinins, and AgNO3 on the efficiency of somatic embryogenesis. After 40 d in culture, the somatic embryos that formed were counted and their
overall growth expressed as pecked cell volume. The selected media consisted of either Gamborg (B5) or Murashige and Skoog
(MS) salts and vitamins supplemented with 40 g 1−1 (117 mM) sucrose, 0.05 mg 1−1 (0.22 μM) 6-benzylaminopurine, and 10 mg l−1 (58.8 μM) AgNO3. Somatic embryo production was substantially reduced at shaker speeds above 40 rpm. Glucose and snerose were the most effective
carbon sources, whereas fructose, galactose, and maltose resulted in a reduced yield and growth of somatic embryos. The development
of somatic embryos was promoted by AgNO3 at concentrations below 10 mg l−1 (58.8 μM). A semi-solid medium containing 1.5 g l−1 Gel-rite produced the highest frequency of somatic embryo conversion, and promoted the efficient growth of plantlets. Using
the reported protocol, over 500 viable somatic embryos were produced per 25 ml of embryogenic cell suspension culture. 相似文献
2.
In the present study an efficient somatic embryogenesis method has been developed in Catharanthus roseus. Friable embryogenic callus was induced from hypocotyl of in
vitro germinated seeds on Murashige and Skoog basal nutrient media supplemented with various auxins particularly 2,4-D (1.0 mg l−1). However, only NAA (1.0 mg l−1) produced somatic embryos in cultures. Embryo proliferation was even high on the same medium added with BAP. Cotyledonary
somatic embryo germinated and converted into plantlets in BAP (0.5 mg l−1) added medium following a treatment with gibberellic acid (1.0 mg l−1) for maturation. Carbon sources and concentrations had a marked influence on maturation process. Plantlet conversion was
better achieved when embryos were matured on 3% fructose or 3–6% maltose. The result discussed in this paper indicates that
somatic embryos were produced in numbers and converted plantlets can be used as raw material, genetic modification to embryo
precursor cell may improve alkaloid yield further. 相似文献
3.
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
4.
Summary Sodium chloride-tolerant plantlets of Dendrocalamus strictus were regenerated successfully from NaCl-tolerant embryogenic callus via somatic embryogenesis. The selection of embryogenic
callus tolerant to 100 mM NaCl was made by exposing the callus to increasing (0–200 mM) concentrations of NaCl in Murashige and Skoog medium having 3% (w/v) sucrose, 0.8% (w/v) agar, 3.0 mg l−1 (13.6 μM) 2,4-dichlorophenoxyacetic acid (2,4-D), and 0.5mg l−1 (2.3μM) kinetin (callus initiation medium). The tolerance of the selected embryogenic callus to 100 mM NaCl was stable through three successive transfers on NaCl-free callus initiation medium. The tolerant embryogenic callus
had high levels of Na+, sugar, free amino acids, and proline but a slight decline was recorded in K+ level. The stable 100 mM NaCl-tolerant embryogenic callus differentiated somatic embryos on maintenance medium [MS medium +3% sucrose +0.8% agar +2.0
mg l−1 (9.0 μM) 2,4-D+0.5 mg l−1 (2.3 μM) kinetin] supplemented with different (0–200 mM) concentrations of NaCl. About 39% of mature somatic embryos tolerant to 100 mM NaCl germinated and converted into plantlets in germination medium [half-strength MS+2% sucrose+0.02 mg l−1 (0.1 μM) α-naphthaleneacetic acid +0.1 mg l−1 (0.49 μM) indole-3-butyric acid] containing 100 mM NaCl. Of these plantlets about 31% established well on transplantation into a garden soil and sand (1:1) mixture containing
0.2% (w/w) NaCl. 相似文献
5.
Jing Li Yang Bo Zhao Eun Soo Seong Myong Jo Kim Won Hee Kang Na Young Kim Chang Yeon Yu Cheng Hao Li 《Plant biotechnology reports》2010,4(4):261-267
We describe culture conditions for a high-efficiency in vitro regeneration system of Papaver
nudicaule through somatic embryogenesis and secondary somatic embryogenesis. The embryogenic callus induction rate was highest when
petiole explants were cultured on Murashige and Skoog (MS) medium containing 1.0 mg l−1 α-naphthaleneacetic acid (NAA) and 0.1 mg l−1 6-benzyladenine (BA) (36.7%). When transferred to plant growth regulator (PGR)-free medium, 430 somatic embryos formed asynchronously
from 90 mg of embryogenic callus in each 100-ml flask. Early-stage somatic embryos were transferred to MS medium containing
1.0 mg l−1 BA and 1.0 mg l−1 NAA to germinate at high frequency (97.6%). One-third-strength MS medium with 1.0% sucrose and 1.0 mg l−1 GA3 had the highest frequency of plantlet conversion from somatic embryos (91.2%). Over 90% of regenerated plantlets were successfully
acclimated in the greenhouse. Secondary somatic embryos were frequently induced directly when the excised hypocotyls of the
primary somatic embryos were cultured on MS medium without PGRs. Sucrose concentration significantly affected the induction
of secondary embryos. The highest induction rate (89.5) and number of secondary somatic embryos per explant (9.3) were obtained
by 1% sucrose. Most secondary embryos (87.2–94.3%) developed into the cotyledonary stage on induction medium. All cotyledonary
secondary embryos were converted into plantlets both in liquid and on semisolid 1/3-strength MS medium with 1.0% sucrose. 相似文献
6.
A simple and efficient protocol is described for regeneration of wild sorghum (Sorghum dimidiatum) from cell suspension cultures. Fast-growing cell suspensions were established from shoot-meristem-derived callus. Plating
of the suspension on Murashige and Skoog agar medium supplemented with 2.5 mg l–1 2,4-dichlorophenoxyacetic acid (2,4-D) resulted in the formation of embryogenic calli. High-frequency (80%) somatic embryogenesis
from small cell clusters (300–400 μm) was observed when the cultures were initially maintained in liquid medium with reduced
levels of 2,4-D (0.25 mg l–1), followed by transfer to regeneration medium. Direct plating of these small clusters on regeneration medium or transfer
to liquid regeneration medium containing kinetin and 6-benzylaminopurine resulted in the development of mature somatic embryos
and plantlets. The regenerants developed to maturity and were all phenotypically and cytologically normal.
Received: 20 May 1998 / Revision received: 1 September 1998 / Accepted: 23 September 1998 相似文献
7.
An efficient somatic embryogenesis system has been established in Catharanthus roseus (L.) G. Don in which primary and secondary embryogenic calluses were developed from hypocotyls and primary cotyledonary somatic
embryos (PCSEs), respectively. Two types of calluses were different in morphology and growth behaviour. Hypocotyl-derived
embryogenic callus (HEC) was friable and fast-growing, while secondary callus derived from PCSE was compact and slow-growing.
HEC differentiated into somatic embryos which proliferated quickly on medium supplemented with NAA (1.0 mg l−1) and BA (1.5 mg l−1). Although differentiation and proliferation of somatic embryos were faster in primary HEC, maturation and germination efficiency
were better in somatic embryos developed from primary cotyledonary somatic embryo-derived secondary embryogenic callus (PCSEC).
At the biochemical level, two somatic embryogenesis systems were different. Both primary and secondary/adventive somatic embryogenesis
and the role of plant growth regulators in two modes of somatic embryo formation have been discussed. 相似文献
8.
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
9.
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
10.
High-frequency plant regeneration via somatic embryogenesis and organogenesis and in vitro flowering of regenerated plantlets in Panax ginseng 总被引:4,自引:0,他引:4
W. Tang 《Plant cell reports》2000,19(7):727-732
The morphogenesis ability of light yellowish globular callus derived from cotyledons of mature zygotic embryos of Panax ginseng was investigated. The optimal media for somatic embryogenesis and shoot organogenesis were MS medium containing 0.5 mg l–1 2,4-dichlorophenoxyacetic acid, 0.1 mg l–1 6-benzyladenine (BA), and 500 mg l–1 lactoalbumin hydrolysate, and SH medium supplemented with 0.5 mg l–1 α-naphthaleneacetic acid, 0.1 mg l–1 BA, and 500 mg l–1casein hydrolysate. The influences of glucose, mannose, fructose, and sorbose in the media on somatic embryogenesis and shoot
organogenesis were revealed as differences in the numbers of somatic embryos and adventitious shoots per gram of morphogenic
callus. The best regeneration of somatic embryos was obtained on medium containing glucose, with a mean of 8.7 somatic embryos
per gram of callus. The best regeneration of shoots was observed on medium containing fructose, with an average of 12.2 adventitious
shoots per gram of callus. Of the somatic embryos 95% were converted into regenerated plantlets, and 100% of adventitious
shoots rooted to form regenerated plantlets. Regenerated plants were successfully established in soil. Flowering was observed
in 5.7% of the regenerated plants derived from shoot organogenesis and in 1.4% of the regenerated plants derived from somatic
embryogenesis.
Received: 1 December 1998 / Revision received: 13 September 1999 / Accepted: 20 September 1999 相似文献
11.
Junaid Aslam Saeed Ahmad Khan Abdul Jaleel Cheruth Abdul Mujib Maheshwar Pershad Sharma Prem Shanker Srivastava 《Saudi Journal of Biological Sciences》2011,18(4):369-380
An efficient somatic embryogenesis system has been established in six date palm (Phoenix dactylifera L.) cultivars (Barhee, Zardai, Khalasah, Muzati, Shishi and Zart). Somatic embryogenesis (SE) was growth regulators and cultivars dependent. Friable embryogenic callus was induced from excised shoot tips on MS medium supplemented with various auxins particularly 2,4-dichlorophenoxyacetic acid (2,4-D, 1.5 mg 1−l). Suspension culture increased embryogenesis potentiality. Only a-naphthaleneacetic acid (NAA, 0.5 mg 1−1) produced somatic embryos in culture. Somatic embryos germinated and converted into plantlets in N6-benzyladenine (BAP, 0.75 mg 1−l) added medium following a treatment with thidiazuron (TDZ, 1.0 mg 1−l) for maturation. Scanning electron microscopy showed early stages of somatic embryo particularly, globular types, and was in masses. Different developing stages of embryogenesis (heart, torpedo and cotyledonary) were observed under histological preparation of embryogenic callus. Biochemical screening at various stages of somatic embryogenesis (embryogenic callus, somatic embryos, matured, germinated embryos and converted plantlets) of date palm cultivars has been conducted and discussed in detail. The result discussed in this paper indicates that somatic embryos were produced in numbers and converted plantlets can be used as a good source of alternative propagation. Genetic modification to the embryo precursor cell may improve the fruit quality and yield further. 相似文献
12.
Embryogenic cultures were initiated and established for the first time in 3 different genotypes of Pinus kesiya using mature zygotic embryos and triacontanol. Mature zygotic embryos produced white-mucilaginous embryogenic callus when
cultured on half strength MSG (Becwar et al. 1990) basal medium supplemented with 90 mM maltose, 2.0 g l−1 Gellan gum, 9.0 M 2, 4-D and 10 g l−1triacontanol. On subculture of such embryogenic callus on the maintenance medium (II) containing 2.0 M 2,4-D and 2.0 g l−1 triacontanol induced cleavage polyembryogenesis with proembryos. The percentage of somatic embryogenesis was not similar
in all the three genotypes. The highest percentage of somatic embryogenesis (88.5 %) was recorded in PK04 genotype. Somatic
embryos were successfully germinated on half strength MSG basal medium without growth regulators. Somatic seedlings showed
fast growth and a survival rate of 95%. This work for the first time reveals that triacontanol can be used as an effective
growth regulator for inducing somatic embryogenesis in conifers. 相似文献
13.
Wei Tang Zhongchen Guo Fan Ouyang 《In vitro cellular & developmental biology. Plant》2001,37(5):558-563
Summary Mature zygotic embryos of eight (open-pollinated) families of loblolly pine (Pinus taeda L.) were cultured on eight different basal salt formulations, each supplemented with 36.2 μM 2,4-dichlorophenoxyacetic acid, 17.8 μM 6-benzyladenine, 18.6 μM kinetin, 500 mg l−1 casein hydrolysate, and 500 mg l−1
l-glutamine for 9 wk; embryogenic tissue was formed on cotyledons, hypocotyls, and radieles of mature zygotic embryos. Callus
was subcultured on the callus proliferation medium, the same as the induction medium but with one-fifth concentration of auxin
and cytokinin for 9 wk. On this medium a white to translucent, glossy, mucilaginous embryogenic callus containing embryogenic
suspensor masses (ESMs) was obtained. The highest frequency of explants forming embryogenic tissue, 17%, occurred on a modified
Murashige and Skoog salts basal medium containing the concentration of KNO3, Ca(NO3)2·4H2O, NH4NO3, KCl, ZnSO4·7H2O, and MnSO4·H2O, 720, 1900, 400, 250, 25.8, and 25.35 mg l−1, respectively. Embryogenic suspension cultures were established by culturing embryogenic callus in liquid callus proliferation
medium. Liquid cultures containing ESMs were transferred to medium containing abscisic acid, polyethylene glycols, and activated
charcoal for stimulating the production of cotyledonary somatic embryos. Mature somatic embryos germinated for 4–12 wk on
medium containing indole-butyric acid, gibberellic acid, 6-benzyladenine, activated charcoal, and reduced sucrose concentration
(15 g l−1). Two hundred and ninety-one regenerated plantlets were transferred to a perlite:peatmoss:vermiculite (1∶1∶1) mixture, then
the plants were transplanted to soil in the earth, and 73 plantlets survived in the field. 相似文献
14.
Summary A procedure for the regeneration of complete plantlets of Tylophora indica from cultured leaf callus via somatic embryogenesis is described. Callus induction from leaf explants was on Murashige and
Skoog (MS) medium with different concentrations of 2,4-dichlorophenoxyacetic acid (2.4-D; 0.03–3 mg l−1; 0.0–13.56 μM) and kinetin (Kn; 0.01 mg l−1; 0.05 μM). The best response for callus induction was obtained on MS medium containing 2 mg l−1 (9.04 μM) 2.4-D and 0.01 mg l−1 (0.05 μM) Kn. After two subeultures on the same medium the embryogenic callus was transferred to MS medium with different concentrations
of the cytokinin, 6-benzyladenine (0.5–3 mg l−1; 2.22–13.32 μM) and 2-isopentenyladenine (2ip; 0.53 mg l−1; 2.46–14.76 μM) along with 0.01 mg l−1 (0.05 μM) indole-3-butyric acid (IBA) for somatic embryo development and maturation. MS medium with 2 mg l−1 (9.84 μM) 2ip produced the maximum number of mature somatic embryos. The mature embryos were bipolar and on transfer to MS basal medium
produced complete plantlets. After hardening the regenerants were planted in the Gudalur forests of Western Ghats. Total DNA
was extracted from 14 regenerants and the mother plant. Random amplified polymorphic, DNA (RAPD) analysis was carried out
using 20 arbitrary oligonucleotides. The amplification products were monomorphic among all the plants revealing the genetic
homogeneity and true-to-type nature of the regenerants. 相似文献
15.
The object of this study was to evaluate different strategies for the production of secondary somatic embryos of cassava on
picloram-supplemented media. Embryogenically competent calli maintained on double-strength Murashige and Skoog (1962) (MS)
medium supplemented with 1 mg l−1 picloram were used as starting material. Secondary embryogenesis from this callus was tested using various basal salt media
in either the solid or the liquid state and containing two different concentrations of picloram. Some of the factors effecting
the conversion of the embryos into plantlets were also studied. A liquid Schenck and Hildebrand (1972) medium containing 60
g l−1 sucrose and 12 mg l−1 picloram favoured the continual production of a highly embryogenic nodular callus. The normal development of somatic embryos
from this tissue was dependant on the use of a picloram-free MS basal salt medium. The embryos were desiccated over a saturated
salt solution of K2SO4 (RH 97.5% at 25 °C) and allowed to develop into plantlets on a MS medium containing 0.1 mg l−1 BA. This procedure allowed for the normal elongation of the embryonic hypocotyl and formation of vigorous and viable shoots
and roots.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
Culture conditions for high frequency plant regeneration via somatic embryogenesis from cell suspension cultures of Ranunculus kazusensis are described. Zygotic embryos formed white nodular structures and pale-yellow calluses at a frequency of 84.9% when cultured
on half-strength Schenk and Hildebrandt (SH) medium supplemented with 0.1 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). However, the frequency of white nodular structure and off-white callus formation
decreased with an increasing concentration of 2,4-D up to 10 mg l−1, when the frequency reached 25%. Cell suspension cultures were established from zygotic embryo-derived pale-yellow calluses
using half-strength SH medium supplemented with 0.1 mg l−1 of 2,4-D. Upon plating onto half-strength SH basal medium, over 90% of cell aggregates gave rise to numerous somatic embryos
and developed into plantlets. Regenerated plantlets were successfully transplanted to potting soil and grown to maturity at
a survival rate of over 90% in a growth chamber. The plant regeneration system established in this study can be applied to
mass propagation and conservation of this species. 相似文献
17.
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
18.
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
19.
A. Othmani C. Bayoudh N. Drira M. Marrakchi M. Trifi 《Plant Cell, Tissue and Organ Culture》2009,97(1):71-79
Plant regeneration through somatic embryogenesis from young leaf explants (5–10 mm long) adjacent to the apex of 5–6 year
old offshoots of Tunisian date palm (Phœnix dactylifera L.), cultivar Boufeggous was successfully achieved. Factors affecting embryogenic callus initiation, including plant growth
regulators and explant size, were investigated. The highest induction frequencies of embryogenic calli occurred after 6–7 months
on MS medium supplemented with 10 mg l−1 2,4-D and 0.3 mg l−1 activated charcoal. The subculture of these calli onto maintenance medium resulted in the formation of proembryos. Fine chopping
and partial desiccation (6 and 12 h) of embryogenic calli with proembryos prior to transfer to MS medium supplemented with
1 mg l−1 ABA stimulated the rapid maturation of somatic embryos. Maturated somatic embryo yield per 0.5 g FW of embryogenic callus
was 51 embryos with an average maturation time of 55 days. This was increased to 422 with finely chopped callus, and 124 and
306 embryos following 6 and 12 h desiccation treatments, respectively. The average time to maturation for these 3 treatments
was 35, 43 and 38 days, respectively. Subsequent substitution of ABA in MS medium with 1 mg l−1 NAA resulted in the germination and conversion of 81% of the somatic embryos into plantlets with normal roots and shoots.
The growth of regenerated somatic plants was also monitored in the field. 相似文献
20.
M. Ramakrishnan S. Antony Ceasar V. Duraipandiyan Melvin A. Daniel S. Ignacimuthu 《In vitro cellular & developmental biology. Plant》2013,49(3):285-293
An efficient somatic embryogenesis and regeneration system was developed for the first time in onion using shoot apex explants. These explants were used to initiate callus in Murashige and Skoog (MS) medium supplemented with 4.0 mg l?1 2,4-dichlorophenoxyacetic acid. The induction frequency of primary callus in this medium was 85.3%. The primary calli were then transferred onto medium supplemented with 2.0 mg l?1 2,4-dichlorophenoxyacetic acid. Following two biweekly subcultures, embryogenic callus formed. Inclusion of a low concentration of 6-benzylaminopurine in the subculture medium promoted the formation of embryogenic callus. The addition of 2.0 mg l?1 glycine, 690 mg l?1 proline, and 1.0 g l?1 casein hydrolysate also increased the frequency of callus induction and embryogenic callus formation. The highest frequency of embryogenic callus (86.9%) and greatest number of somatic embryos (26.3 per callus) were obtained by the further addition of 8.0 mg l?1 silver nitrate. Somatic embryos formed plantlets on regeneration medium supplemented with 1.5 mg l?1 6-benzylaminopurine; addition of 2.0 mg l?1 glycine to the regeneration medium promoted a high frequency of regeneration (78.1%) and plantlet formation (28.7 plants per callus). The regenerated plantlets were transferred to half-strength MS medium supplemented with 1.5 mg l?1 indole-3-butyric acid for root development; the maximum frequency of root formation was 87.7% and the average number of roots was 7.6 per shoot. The regenerated plantlets were successfully grown to maturity after hardening in the soil. This is the first report of somatic embryogenesis and regeneration from shoot apex explants of onion. 相似文献