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1.
Iyyakkannu Sivanesan Mi Young Lim Byoung Ryong Jeong 《Plant Cell, Tissue and Organ Culture》2011,107(2):365-369
A simple and efficient protocol was developed for somatic embryogenesis from leaf and petiole explants of Campanula punctata Lam. var. rubriflora Makino. Somatic embryos (SE) were obtained with greater frequency from petiole explants than from leaf explants when cultured
on Murashige and Skoog (MS) medium supplemented with 2.0 mg L−1 2,4-dichlorophenoxyacetic acid (2,4-D) and 1.0 mg L−1 6-benzyladenine (BA). On this medium, a mean number of 19.5 and 31.2 SE were developed per leaf and petiole explants, respectively.
Embryos were induced both light and dark conditions but culturing the explants 2 weeks in the dark followed by 3 weeks under
light resulted in high frequency of embryo formation. Globular embryos germinated best on MS medium supplemented with 0.3%
(w/v) activated charcoal (AC) and 1.0 mg L−1 GA3. The germinated plantlets grew further on MS medium containing 0.3% AC. Plantlets were successfully acclimatized in the greenhouse
with 94% survival rate. This is the first report on induction of somatic embryogenesis in this genus and also has implications
for genetic transformation, and mass clonal propagation. 相似文献
2.
Summary
In vitro regeneration of plants via somatic embryogenesis through cell suspension culture was achieved in horsegram. Embryogenic calluses
were induced on leaf segments on solid Murashige and Skoog (MS) medium with 9.0 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Differentiation of somatic embryos occurred when the embryogenic calluses were transferred
to liquid MS medium containing 2,4-D. Maximum frequency (33.2%) of somatic embryos was observed on MS medium supplemented
with 7.9 μM 2,4-D. Cotyledonary-torpedo-shaped embryos were transferred to liquid MS medium without growth regulators for maturation
and germination. About 5% of the embryos germinated into plants, which grew further on solid MS medium. The plants were hardened
and established in soil. Effects of various auxins, cytokinins, carbohydrates, amino acids, and other additives on induction
and germination of somatic embryos were also studied. A medium supplemented with 7.9 μM 2,4-D, 3.0% sucrose, 40 mg l−1
L-glutamine, and 1.0 μM abscisic acid was effective to achieve a high frequency of somatic embryo induction, maturation, and further development. 相似文献
3.
Ai Hua Chen Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,102(3):357-364
We developed a new protocol for highly efficient somatic embryogenesis and plantlet conversion of Schisandra chinensis. Friable embryogenic callus was induced from cotyledonary leaves and hypocotyls of germinated zygotic embryos on Murashige
and Skoog (MS) agar medium containing 2,4-dichlorophenoxyacetic acid (2,4-D). Preculture of zygotic embryos on 2,4-D-containing
medium increased embryogenic callus induction efficiency. The highest embryogenic callus induction frequency of 56.7% was
obtained from shoot apical meristem-containing hypocotyl explants from 1-week-old germinated embryos on MS medium containing
4.0 mg l−1 2,4-D. Embryogenic callus proliferation, somatic embryo (SE) formation, and subsequent plantlet conversion occurred under
optimal culture conditions. The effects of MS medium strength, sucrose, gibberellic acid (GA3), and 6-benzyladenine (BA) on SE formation and plantlet conversion were evaluated. Low MS medium strength (1/4 to 1/2) was
necessary for SE formation, and the optimal sucrose concentration was 2.0%. Supplementing medium with GA3 negatively impacted SE formation and subsequent development. BA significantly increased the number of SEs and the plantlet
conversion capacity. One-third-strength MS medium with 1.0% sucrose and 0.5 mg l−1 BA produced the highest number of SEs (309 embryos from 9 mg embryogenic callus) and the highest frequency of plantlet conversion
from germinated SEs (52.6%). When transplanted to soil, 90% of the regenerated plants developed into normal plants. 相似文献
4.
Five genotypes of chickpea (Cicer arietinum L.) PG1, PG5, PG12, N59 and C235 were evaluated for induction of somatic embryogenesis. Somatic embryogenesis was induced from immature cotyledons of genotypes PG12 and C235 and immature embryo axes of genotypes PG5, PG12 and C235. Genotypes N59 and PG1 showed no response. The maximum frequency of globular embryo formation occurred in cotyledonary segments on MS medium with 3.0 mg/l 2,4,5-trichlorophenoxyacetic acid (2,4,5-T). Further embryo development was achieved only in somatic embryos derived from cotyledonary segments of genotype PG12. Globular-stage embryos derived from immature embryo axes of PG5, C235, PG12, and cotyledonary segments of C235 dedifferentiated and formed callus. The cotyledonary stage embryos of genotype PG12 germinated on half-strength MS medium supplemented with 1 mg/l zeatin. The regenerated plants were transferred to soil and grown to maturity.Abbreviations ABA
abscisic acid
- BAP
6-benzylamino purine
- CM
coconut milk
- Dicamba
3,6-Dichloro-2-methoxybenzoic acid
- 2,4-D
2,4-dichlorophenoxy-acetic acid
- GA3
gibberellic acid
- IAA
indoleacetic acid
- MS
Murashige and Skoog medium (1962)
- NAA
1-napthaleneacetic acid
- Picloram
4 amino-3,5,6-trichloropicolinic acid
- 2,4,5-T
2,4,5-trichlorophenoxy-acetic acid
- zeatin
(6-[4-Hydroxy-3-methyl-2-butenylamino] purine) 相似文献
5.
Somatic embryogenesis of <Emphasis Type="Italic">Myrciaria aureana</Emphasis> (Brazilian grape tree)
Sergio Yoshimitsu Motoike Edson Santana Saraiva Marilia Contin Ventrella Crislene Viana Silva Luiz Carlos Chamhum Salomão 《Plant Cell, Tissue and Organ Culture》2007,89(1):75-81
The aim of this research was to establish a long-term somatic embryogenic cultures that could be used for cryopreservation.
For the induction of somatic embryogenesis, different levels of 2,4-D as well as the combination of 2,4-D and indole-3-acetyl-l-aspartic acid (IASP) were tested on cotyledons of zygotic embryos. The somatic embryogenic cultures were established and
maintained up to 2 years through frequent subculturing on a medium containing 2,4D + IASP. Light, activated charcoal, and
polyethylene glycol (PEG) were tested for the regeneration and maturation of somatic embryos, and the mature embryos were
germinated in JADS medium. The combination of light and PEG provided the highest number of mature embryos. The somatic embryos
obtained were smaller than zygotic embryos and lacked starch. There was an interaction between 2,4-D and IASP on the induction
and regeneration of somatic embryo in Myrciaria aureana. The combination of light and PEG increased the number of mature embryos; however, charcoal was detrimental to the process. 相似文献
6.
Immature and mature zygotic embryos of Paspalum scrobiculatum L. cv. PSC 1 cultured on MS or N6 nutrient medium supplemented with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D), formed embryogenic callus. Induction of embryogenic callus and subsequent somatic embryogenesis was possible at a lower concentration of 2,4-D on N6 than MS medium. Immature embryos were highly totipotent, forming somatic embryos at a higher frequency than mature embryos. Addition of amino acids (L-proline or L-tryptophan) to 2,4-D medium resulted in significant enhancement of embryogenesis on culture of mature embryos. Silver nitrate also supported an increased frequency of embryogenesis. Thus it is possible to have high frequency of somatic embryogenesis on culture of mature embryos, which are available in abundance and with ease than immature embryos. The somatic embryos readily germinated and formed plantlets on hormone-free regeneration medium. The regenerated plantlets were successful on transfer to soil and set seed. 相似文献
7.
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and petiole expiants has been developed in sweetpotato [Ipomoea batatas L. (Lam.)]. The optimal somatic embryogenic response was obtained in the genotype PI 318846-3 with a two-step protocol: (1) stage I-incubation of expiants in the dark for 2 weeks on Murashige Skoog (MS) medium containing 2,4-dichlorophenoxyacetic acid (2,4-D) (2.5 mg/l) and 6-benzylaminopurine (0.25 mg/l) and, (2) stage II-culture in the light on MS medium with abscisic acid (ABA) (2.5 mg/l). The addition of ABA was critical for enhanced production of somatic embryos. Secondary somatic embryos were produced from the primary embryos cultured on MS medium with 2,4-D at 0.2 mg/l. The somatic embryos were converted into normal plantlets when cultured on basal MS medium. Upon transfer to soil, plants grew well and appeared normal with no mortality. The system of somatic embryogenesis described here will facilitate tissue culture, germplasm conservation and gene transfer research of sweetpotato due to its rapidity (6 to 10 weeks), prolific plant production by direct embryogenesis, ease of secondary somatic embryo production and reproducibility.Abbreviations ABA
abscisic acid
- BAP
6-benzylaminopurine, 2,4-D-2,4-dichlorophenoxyacetic acid
- GA3
gibberellic acid
- KIN
kinetin
- MS
medium of Murashige and Skoog (1962)
- NAA
1-naph-thaleneacetic acid
- PIC
picolinic acid
- TDZ
thidiazuron 相似文献
8.
Summary Four auxins (2,4-dichlorophenoxyacetic acid [2,4-D], indole-3-acetic acid [IAA], indole-3-butyric acid [IBA], and naphthaleneacetic
acid [NAA]), and five cytokinins (N
6-[2-isopentenyl]-adenine [2iP], N
6-benzyladenine [BA], 6-furfurylaminopurine [kinetin], 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea [TDZ], and 6-[4-hydroxy-3-methylbut-2-enylamino]purine
[zeatin]) were examined for their effects on direct embryo induction from leaf explants of Dendrobium cv. Chiengmai Pink cultured on 1/2 Murashige and Skoog (MS) medium. Whether in light or darkness, explants easily became
necrotic and no embryos were obtained on growth regulator-free or auxin-containing media after 60 d of culture. By contrast,
five cytokinins tested induced direct embryo formation from leaf explants, and explants cultured in light had a higher embryogenic
response compared with those cultured in darkness. The best condition for direct embryo induction was at 18.16 μM TDZ cultured in light for 60 d, where 33% of explants formed a mean number of 33.6 embryos per explant. During subculture
on growth regulator-free 1/2 MS medium, embryos gradually developed into plantlets. Secondary embryogenesis was occasionally
found on sheath leaves of embryos. Regenerated plantlets were successfully transplanted and grown in a greenhouse environment. 相似文献
9.
The effect of auxins, time exposure to auxin and genotypes on somatic embryogenesis from mature embryos of wheat 总被引:3,自引:0,他引:3
Mikhail Filippov Dmitry Miroshnichenko Darya Vernikovskaya Sergey Dolgov 《Plant Cell, Tissue and Organ Culture》2006,84(2):100192-100201
The effects of different factors on the embryogenesis and plant regeneration from mature embryos of Russian spring and winter
genotypes were studied. Embryogenic callus induction was achieved on MS medium supplemented with different concentrations
of 2,4-D (2,4-dichlorophenoxyacetic acid), 2,4,5-T (2,4,5-trichlorophenoxyacetic acid) or Dicamba (3,6-dichloro-o-anisic acid). Although all auxins were able to induce callus from explants with high frequency (98–100%), Dicamba was more
effective for the induction of embryogenic callus (21.8–38.3%). Maximum embryogenic callus formation and high number of regenerated
plants were observed at 12 mg l−1 of Dicamba. The time exposure to Dicamba (7, 14, 21 and 28 days) had a significant effect on efficiency of somatic embryogenesis.
When contact of explants with callus induction medium was increased from 7 to 21 days the rate of somatic embryogenesis and
number of regenerated plants per embryogenic callus gradually increased from 13.0 to 38.4% and 3.6 to 8.0%, respectively.
Supplement of additional auxins (indoleacetic acid (IAA), indolebutyric acid (IBA), and naphthaleneacetic acid (NAA)) to callus
induction medium with Dicamba had a positive effect on the rate of embryogenic callus formation, while the average number
of regenerated shoots was not affected. The best rate of somatic embryogenesis was observed at the addition of 0.5 mg l−1 IAA with Dicamba (61.0%). The optimum combination of Dicamba and IAA increased the efficiency of somatic embryogenesis and
plant regeneration from seven spring and winter wheat genotypes, thought overall morphogenic capacity was still genotype dependent. 相似文献
10.
Summary We report a protocol for somatic embryogenesis and plantlet regeneration of Buchanania lanzan Spreng (Family—Anacardiaceae), which is a tropical fruit tree widely distributed in the dry forests of India. Calluses were initiated from immature zygotic
embryos cultured on Murashige and Skoog (MS) medium supplemented with various combinations of 2,4-dichlorophenoxyacetic acid
(2,4-D), 6-benzyladenine (BA) and/or 1-naphthaleneacetic acid (NAA). The highest frequency (60%) of somatic embryo induction
was obtained in cultures grown on MS medium fortified with 4.53 μM 2,4-D, 5.32 μM NAA and 4.48 μM BA. The medium supplemented with 15 μM abscisic acid (ABA) was most effective for maturation and germination of somatic embryos. This is the first report on somatic
embryogenesis in B. lanzan, which may be helpful for in vitro propagation, ex situ conservation and genetic manipulation of this species. 相似文献
11.
Ling Yang Jianan Wang Lei Bian Yuhua Li Hailong Shen 《Plant Cell, Tissue and Organ Culture》2012,111(2):173-182
An efficient protocol for secondary somatic embryogenesis in mountain ash is reported. Secondary somatic embryos (SSEs), initially obtained from primary embryos, were proliferated and maintained for more than 2?years via cyclic secondary somatic embryogenesis. SSEs were produced on the surfaces of cotyledons and radicles of maternal somatic embryos. Histological observations of the various stages of SSE development revealed four typical stages: globular, heart-shaped, torpedo, and cotyledon. Addition of a low concentration of naphthaleneacetic acid (NAA) to Murashige and Skoog (MS) medium resulted in the induction of SSEs, but addition of 2,4-dichlorophenoxyacetic acid (2,4-d) to MS medium decreased SSE formation. Addition of casein hydrolysate (CH) to MS medium promoted induction of SSEs. Cotyledonary SSEs were cultured on MS medium with 20?C60?g?L?1 sucrose under light for 1?month until maturation. After transferred to MS medium containing either 0.06???M NAA or 0.15???M indole-3-butyric acid in the light, over 50?% of the mature SSEs developed into plantlets. Addition of 1.0?g?L?1 activated charcoal was beneficial for SSE germination (over 60?%). At 18?°C, over 90?% of the germinated SSEs converted to plantlets on ? MS (half-strength of MS macroelements) with 1.8???M NAA under light. Plantlets acclimatized successfully to ex vitro conditions and field plants developed with normal phenotypes. 相似文献
12.
Somatic embryos were induced on peanut (Arachis hypogaea) leaflets from aseptically germinated embryo axes. Leaflet size influenced percent somatic embryogenesis; 5–8 mm long cut leaflets were superior to 2–3 mm long uncut leaflets. Maximum embryogenesis of 14.6% was obtained after a 15 d incubation on induction medium (modified MS with B5 vitamins, 30 g/l sucrose, 4 g/l Gel-Gro, 40 mg/l 2,4-D +0.2 mg/l kinetin) followed by transfer to a secondary medium with 5 mg/l 2,4-D+0.2 mg/l kinetin. Primary somatic embryos were fused along the axes with no distinct cotyledons, but secondary embryos had single axes with two cotyledons. Other treatments had lower percent embryogenesis, no secondary embryogenesis, and embryos with single axes with two cotyledons. Some somatic embryos converted into normal plants capable of greenhouse survival.Abbreviations MS
Murashige and Skoog (1962) medium
- B5
Gamborg et al. (1968) B5 medium
- 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6benzylaminopurine
- NAA
1-naphthaleneacetic acid 相似文献
13.
J. L. Yang E. S. Seong M. J. Kim B. K. Ghimire W. H. Kang Chang Yeon Yu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2010,100(1):49-58
Cotyledon, hypocotyl or root explants of 7-day-old broccoli seedlings were cultured on Murashige and Skoog (MS) agar or liquid
medium supplemented with 1.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). The frequency of direct somatic embryo formation was 100% when root explants were
cultured in liquid medium. Histological analysis indicated that somatic embryos were initiated directly from the pericycle
cell layers of root explants as early as 1 day after liquid culture. Genotype did not affect the frequency of somatic embryo
formation or the number of somatic embryos per explant. All broccoli genotypes examined had 100% somatic embryo induction
efficiency, and the number of somatic embryos per 0.8 mm root segment ranged from 22.9 in ‘Luhui’ to 26.0 in ‘Haizi’. The
number of normally developed somatic embryos in culture increased with increasing 2,4-D concentration. Plantlet regeneration
frequency was the highest (73.3%) when germinated plantlets were transferred to 1/2 strength MS agar medium containing 1.0 mg l−1 6-benzyladenine (BA). When regenerated plantlets were transferred to a greenhouse, approximately 75% survived and there were
no morphological differences between regenerated plants and seed-derived controls. The protocols established in this study
will benefit large-scale vegetative propagation and transformation-based genetic improvement of broccoli. 相似文献
14.
Jing Li Yang Yu Da Niu Chuan Ping Yang Gui Feng Liu Cheng Hao Li 《Plant Cell, Tissue and Organ Culture》2011,106(3):391-399
Somatic embryogenesis (SE) was induced in female flower buds from mature Schisandra chinensis cultivar ‘Hongzhenzhu’. Somatic embryo structures were induced at a low frequency from unopened female flower buds and excised
unopened on Murashige and Skoog (MS) agar medium containing 4.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D). Friable embryogenic calli were induced from somatic embryo structures after three
to four subcultures on initiation medium. The frequencies of mature somatic embryo germination and plantlet conversion were
low, but increased in the presence of gibberellic acid (GA3). Some germinated somatic embryos could form friable embryogenic calli on medium without plant growth regulators (PGRs).
The germination and conversion frequencies of somatic embryos from embryogenic calli induced using PGR-free medium were higher
than for somatic embryos from embryogenic calli induced on medium containing 2,4-D. Most somatic embryos from 2,4-D-induced
embryogenic calli had trumpet-shaped embryos, and most somatic embryos from PGR-free medium–induced embryogenic calli had
two or three cotyledons. Histological observation indicated that two- and three-cotyledon embryos had defined shoot primordia,
but most of the trumpet-shaped embryos yielded plantlets that lacked or had poorly developed meristem tissue. Cytological
and random amplification of polymorphic DNA (RAPD) analyses indicated no evidence of genetic variation in the plantlets of
somatic embryo origin. 相似文献
15.
Rapid propagation of Eleutherococcus senticosus via direct somatic embryogenesis from explants of seedlings 总被引:5,自引:0,他引:5
Explants from three different parts (cotyledon, hypocotyl or root) of one week-old seedlings of Eleutherococcus senticosus were cultured on Murashige and Skoog (MS) medium with 1.0 mg l-1 2,4-D. Somatic embryos were formed directly from the surfaces of explants. The frequency of direct somatic embryo formation
was the highest in the hypocotyl segments (75%) as compared to cotyledon (56%) or root segments (12%). When hypocotyl explants
from 3 different stages of seedlings (zero, one or three week-old) were cultured on MS medium with 1.0 mg l-1 2,4-D, the frequency of somatic embryo formation rapidly declined as the zygotic embryos germinated. However most somatic
embryos (93%) from explants of zygotic embryos developed as fused state (multiple embryo), whereas somatic embryos (over 89%)
from more developed seedlings developed into single state (single embryo). Single embryos germinated and regenerated into
plantlets with both shoots and roots, while multiple embryos only regenerated into only multiple shoots. Plantlets that regenerated
from single embryos of E. senticosus were acclimatized in a greenhouse.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
16.
Plant regeneration via somatic embryogenesis was achieved in callus cultures derived from nodal explants of in vitro grown seedlings and excised mature zygotic embryos of three bamboo species on Murashige and Skoog's (MS) basal medium supplemented with 0.5 mg/l kinetin (Kn), 2.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D), 10 mg/l adenine sulphate (Ads) and 3% (w/v) sucrose incubated in the light or in the dark. Somatic embryos germinated (95–98%) into normal plants and were transferred to soil with 95% success. In vitro flowering was induced on shoots developed from nodal explants taken from somatic embryo regenerated plants of Bambusa vulgaris, Dendrocalamus giganteus and Dendrocalamus strictus on half-strength MS basal medium supplemented with 0.25 mg/l indole-3-butyric acid (IBA), 0.5 mg/l Ads, 0.5 mg/l gibberellic acid (GA3) and 3% sucrose.Abbreviations BAP
6-benzylaminopurine
- Kn
kinetin
- Ads
adenine sulphate
- IBA
indole-3-butyric acid
- NAA
1-naphthaleneacetic acid
- 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) basal medium
- GA3
gibberellic acid 相似文献
17.
C. P. Kaviraj G. Kiran R. B. Venugopal P. B. Kavi Kishor Srinath Rao 《In vitro cellular & developmental biology. Plant》2006,42(2):134-138
Summary This study reports a protocol for plant regeneration from cultured explants of green gram [Vigna radiata (L.) Wilezek] via somatic embryogenesis. Somatic embryos were induced from nature cotyledons of var., TAP-7 and Pusa Baisaki
when cultured on Murashige and Skoog (MS) medium fortified with different concentrations of 2,4-dichlorophenoxyacetic acid,
α-naphthaleneacetic acid (NAA) and 2,4,5-trichlorophenoxyacetic acid singly or in combination with 2.22–8.88 μM N
6-benzylaminopurine (BAP) or 2.32–9.38 μM kinetin. The type and concentration of auxin and plant genotype influenced the frequency of somatic embryogenesis. NAA was
the most effective auxin for somatic embryo induction. The well-developed, cotyledonary shaped embryos of var. TAP-7 germinated
into plantlets at a frequency of 56.6% on MS medium supplemented with 1.88 μM abscisic acid and 6.66 μM BAP. Regenerated plants were transferred to soil and grown to maturity with 90% survival. The protocol described here offers
a good potential for genetic improvement using gene transfer techniques and the production of synthetic seeds of V. radiata. 相似文献
18.
Summary
In vitro propagation of Andrographis paniculata (Burm. f.) Wallich ex Nees through somatic embryogenesis, and influence of 2,4-dichlorophenoxyacetic acid (2,4-1) on induction, maturation, and
conversion of somatic embryos were investigated. The concentration of 2,4-D in callus induction medium determined the induction,
efficacy of somatic embryogenesis, embryo maturation, and conversion. Friable callus initiated from leaf and internode explants
grown on Murashige and Skoog (MS) medium supplemented with 2.26, 4.52, 6.78, and 9.05μM 2,4-D started to form embryos at 135, 105, 150, and 185d, respectively, after explant establishment. Callus initiated at
13.56μM 2,4-D did not induce embryos even after 240 d, whereas those initiated on MS medium with 4.52μM 2,4-D was most favorable for the formation and maturation of somatic embryos. Callus subcultured on the medium with reduced
concentration of 2,4-D (2.26μM) became embryogenic. This embryogenic callus gave rise to the highest number of embryos (mean of 312 embryos) after being
transferred to half-strength MS basal liquid medium. The embryos were grown only up to the torpedo stage. A higher frequency
of embryos developed from callus initiated on 2.26 or 4.52 μM 2,4-D underwent maturation compared to that initiated on higher concentrations of 2.4-D. The addition of 11.7μM silver nitrate to half-strength MS liquid medium resulted in 71% of embryos undergoing maturation, while 83% of embryos developed
into plantlets after being transferred to agar inedium with 0.44 μMN6-benzyladenine and 1.44 μM gibberellic acid. Most plantlets (88%) survived under field conditions and were morphologically identical to the parent plant. 相似文献
19.
Somatic embryogenesis was obtained from cotyledon and mature zygotic embryo callus cultures of Terminalia chebula Retz. Callus cultures of cotyledon and mature zygotic embryo were initiated on induction medium containing Murashige and Skoog (MS) nutrients with 1.0 mg/l 2,4-dichlorophenoxyacetic acid (2,4-D) either 0.01 or 0.1 mg/l Kinetin and 30 g/l sucrose. Induction of somatic embryogenesis, proliferation and development was obtained through different culture passages. Embryogenic cotyledon callus with globular somatic embryos was obtained on MS basal medium supplemented with 50 g/l sucrose. Globular somatic embryos were observed from mature zygotic embryo callus on induction medium. Different stages of somatic embryo development from cotyledon and mature zygotic embryo calluses were observed on MS basal medium supplemented with 50 g/l sucrose after 4 weeks of culture. Histological studies have revealed the developmental stages of somatic embryos. A maximum of 40.3±1.45 cotyledonary somatic embryos/callus was obtained from mature zygotic embryo compared to 7.70±0.37 cotyledonary somatic embryos/callus initiated from cotyledons. Germination of somatic embryos and conversion to plants were achieved. Highest frequency of germination (46.66±0.88) of somatic embryos was obtained on MS basal medium containing benzyladenine (0.5 mg/l) with 30 g/l sucrose. 相似文献
20.
Jin Cui Jianjun Chen Richard J. Henny 《In vitro cellular & developmental biology. Plant》2009,45(1):34-43
Plant regeneration through direct somatic embryogenesis in Aeschynanthus radicans ‘Mona Lisa’ was achieved in this study. Globular somatic embryos were formed directly from cut edges of leaf explants and
cut ends or on the surface of stem explants 4 wk after culture on Murashige and Skoog (MS) medium supplemented with N-phenyl-N′-1, 2, 3-thiadiazol-5-ylurea (TDZ) with α-naphthalene acetic acid (NAA), TDZ with 2,4-dichlorophenoxyacetic acid (2,4-D),
or 6-benzylaminopurine (BA) or kintin (KN) with 2,4-D. MS medium containing 9.08 μM TDZ and 2.68 μM 2,4-D resulted in 71%
of stem explants producing somatic embryos. In contrast, 40% of leaf explants produced somatic embryos when induced in medium
containing 6.81 μM TDZ and 2.68 μM 2,4-D. Somatic embryos matured, and some germinated into small plants on the initial induction
medium. Up to 64% of stem explants cultured on medium supplemented with 9.08 μM TDZ + 2.68 μM 2,4-D, 36% of leaf explants
cultured on medium containing 6.81 μM TDZ and 2.68 μM 2,4-D had somatic embryo germination before or after transferring onto
MS medium containing 8.88 μM BA and 1.07 μM NAA. Shoots elongated better and roots developed well on MS medium without growth
regulators. Approximately 30–50 plantlets were regenerated from each stem or leaf explant. The regenerated plants grew vigorously
after transplanting to a soil-less substrate in a shaded greenhouse with more than a 98% survival rate. Three months after
their establishment in the shaded greenhouse, 500 plants regenerated from stem explants were morphologically evaluated, from
which five types of variants that had large, orbicular, elliptic, small, and lanceolate leaves were identified. Flow cytometry
analysis of the variants along with the parent showed that they all had one identical peak, indicating that the variant lines,
like the parent, were diploid. The mean nuclear DNA contents of the variant lines and their parent ranged from 4.90 to 4.99 pg
2C−1, which were not significantly different statistically. The results suggest that the regenerated plants have a stable ploidy
level, and the regeneration method established in this study can be used for rapid propagation of ploidy-stable Aeschynanthus radicans. 相似文献