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1.
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
2.
Katharina Hristoforoglu Josef Schmidt Harald Bolhar-Nordenkampf 《Plant Cell, Tissue and Organ Culture》1995,40(3):277-284
Mature zygotic embryos of Abies alba mull were placed on a modified MCM medium (basal medium-BM) with 2.2 M benzyladenine and 2.3 M kinetin to induce embryogenic suspensor masses (ESM). These ESM proliferated on induction medium supplemented with 0.2 M 2,4-dichlorophenoxyacetic acid. From 61 ESM lines induced, 36 are still in culture after 2 years, of which 18 show embryogenic potential indicated by spontaneous formation of globular somatic embryos on the proliferation medium supplemented with 500–1000 mg l-1 casein hydrolysate and 500 mg l-1
l-glutamine. ESMs from cell line 2/56 were conditioned 1 week on BM with 58 mM sucrose and 10 g l-1 activated charcoal for maturation of somatic embryos. Maturation was achieved on BM containing 20 M (±)cis-trans-abscisic acid in combination with 111 mM maltose. Organic nitrogen supplements improved the proliferation rate of cell line 2/56 as well as the maturation and vitality of the somatic embryos. Partial drying was necessary for subsequent root development. Plantlets with a root, primary needles and a terminal bud developed on BM when a combination of 30 mM sucrose and 50 mM maltose was provided as carbon source.Abbreviations BM
basal medium
- BA
benzyladenine
- ESM
embryogenic suspensor mass
- 2,4-d
2,4-dichlorophenoxyacetic acid
- CH
casein hydrolysate
-
l-gln
l-glutamine
- ABA
(±) cis-trans-abscisic acid 相似文献
3.
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
4.
Three pumpkin embryogenic lines were initiated on wounded zygotic embryos cultured on medium with or without 2,4-dichlorophenoxyacetic acid (2,4-D). Somatic embryo development was controlled by the availability of various compounds in the medium: presence/absence of 2,4-D, nitrogen sources. The highest rate of DNA methylation was in the early embryo stages, predominantly on MSC medium with 2,4-D and on auxin-free medium supplemented with 1.0 mM NH4Cl. DNA methylation was correlated with early embryo development in a manner that was not exclusively dependent on the presence/absence of exogenous auxin. DNA methylation decreased during embryo maturation on auxin-free MSC medium and on auxin-free MSC supplemented with 12.3 M 5-azacytidine (5-azaC). The embryogenic features of the pumpkin tissue were preserved, even after a 2-month treatment with 5-azaC.Abbreviations 5-azaC
5-Azacytidine
- CRED-RA
Coupled restriction enzyme digestion and random amplification
- 2,4-D
2,4-Dichlorophenoxyacetic acid
- DNMRT
Duncans new multiple range test
- IAA
Indole-3-acetic acid
- 5-mC
5-Methylcytosine 相似文献
5.
Summary Efficient in vitro propagation of Ceropegia candelabrum L. (Asclepidaceae) through somatic embryogenesis was established. Somatic embryogenesis depended on the type of plant growth
regulators in the callus-inducing medium. Friable callus, developed from leaf and internode explants grown on Murashige and
Skoog (MS) medium supplemented with 4.52μM2,4-dichlorophenoxyacetic acid (2,4-D), underwent somatic embryogenesis. Compared to solid media, suspension culture was superior
and gave rise to a higher number of somatic embryos. Transfer of the friable callus developed on MS medium containing 4.52μM 2,4-D to suspension cultures of half- or quarter-strength MS medium with lower levels of 2,4-D (0.23 or 0.45 μM) induced the highest number of somatic embryos, which developed up to the torpedo stage. Somatic embryogenesis was asynchronous
with the dominance of globular embryos. About 100 mg of callus induced more than 500 embryos. Upon transfer to quarter-strength
MS agar medium without growth regulators, 50% of the somatic embryos underwent maturation and developed into plantlets. Plantlets
acclimatized under field conditions with 90% survival. 相似文献
6.
Summary Regeneration of plants via somatic embryogenesis was achieved from zygotic embryo explants isolated from mature seeds of Schisandra chinensis. Merkle and Sommer's medium, fortified with 2,4-dichlorophenoxyacetic acid (2,4-D; 9.04 μM) and zeatin (0.09 μM), was effective for induction of embryogenic callus. The development of a proembryogenic mass and somatic embryos occurred
on Murashige and Skoog medium (MS) free of plant growth regulators. The embryogenic callus induced on Merkle and Sommer's
medium supplemented with 2,4-D (9.04 μM) and zeatin (0.09 μM) showed development of the maximum number of somatic embryos when transferred to MS medium free of plant growth regulators.
The maximum maturation and germination of cotyledonary somatic embryos (46.3%) occurred on MS medium supplemented with 2,4-D
(0.45 μM) and N6-benzyladenine (1.11 μM). The somatic embryo-derived plants were successfully hardned, with a survival rate of approximately 67%, and established
in the field. 相似文献
7.
Veena Agrawal Pratima Rani Sardar 《In vitro cellular & developmental biology. Plant》2007,43(6):585-592
In vitro regeneration through somatic embryogenesis as well as organogenesis using cotyledon of a woody medicinal legume, Cassia angustifolia is reported. The cotyledons dissected from semi-mature seeds, if inoculated on Murashige and Skoog’s medium (MS) supplemented
with auxin alone or in combination with cytokinin, produced direct and indirect somatic embryos. A maximum of 14.36 ± 2.26
somatic embryos per 20 mg of explants including callus were produced in 70% cultures on MS medium with 2.5 μM benzyladenine
(BA) + 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Although the percentage of embryogenic cultures was higher (83.33%) at
10 μM 2,4-D + 1 μM BA, the average number of somatic embryos was much less (7.6 ± 0.85) at this level, whereas at 2.5 μM BA
and 5 μM 2,4-D, there was a simultaneous formation of both somatic embryos and shoots. The somatic embryos, although started
germinating on the same medium, developed into full plantlets only if transferred to MS basal with 2% sucrose. Cytokinins
alone did not induce somatic embryogenesis, but formed multiple shoots. Five micromolar BA proved optimum for recurrently
inducing shoots in the competent callus with a maximum average of 12.04 ± 2.10 shoots and shoot length of 2.26 ± 0.03 cm.
Nearly 91.6% shoots (2–2.5 cm in size) organized an average of 5.12 ± 0.58 roots on half strength MS + 10 μM indole-3-butyric
acid. All the plantlets have been transferred successfully to soil. Types of auxin and its interaction with cytokinin significantly
influenced somatic embryogenesis. 相似文献
8.
Efficient plant regeneration via somatic embryogenesis has been developed in chickpea cultivar C235. Leaf explants, on MS medium supplemented with 1.25 mg/l 2,4-D and 0.25 mg/l kinetin, yielded somatic embryos with high efficiency during dark incubation. MS medium supplemented with B5 vitamins, 0.125 mg/l IBA and 2 mg/l BAP was found suitable for embryo maturation. The well formed embryos germinated into plantlets on basal B5 medium supplemented with 0.25 mg/l BAP. Further development into healthy plantlets was obtained on basal B5 medium. Hardened plantlets produced normal, fertile plants upon transfer to soil.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-Benzyl-aminopurine
- IAA
IndoIe-3-acetic acid
- IBA
Indole-3-butyric acid
- NAA
1-Naphthalene acetic acid
- Kinetin
6-furfuryl aminopurine
- Zeatin
6-(4-hydroxy-3-methylbut-2-enylamino)-purine 相似文献
9.
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
10.
Somatic embryogenesis (SE) was successfully induced from mature zygotic embryos of seven families of Picea likiangensis (Franch.) Pritz after 20 weeks culture on initiation medium. Three basal media (one-half strength LM medium, one-half strength
LP medium and improved LP medium) with different concentrations of 2,4-dichlorophenoxyacetic acid (2,4-D) and 6-benzyladenine
(6-BA) were tested but only one-half strength LM medium supplemented with 2,4-D and 6-BA was successful for the embryogenic
cultures (EC) initiation. The initiation frequencies of EC varied greatly from different families when culturing on the same
initiation medium. The highest frequency (41.3%) was induced from one of the families on one-half strength LM medium supplemented
with 3 mg L−1 2,4-D and 1.5 mg L−1 6-BA and 16.83% on average for seven families. EC were subcultured and proliferated on the same medium as the initiation
one every 10 days. 3 lines of EC induced from the same family were applied in maturation experiment. Cotyledonary somatic
embryos were observed after EC were transferred to maturation media of one-half strength LM medium containing 20-80 mg L−1 abscisic acid and 7.5% polyethylene glycol (PEG-4000). However, one-half strength LM medium supplemented with 40 mg L−1 or 60 mg L−1 ABA and 7.5% PEG gave the best maturation and the 3 lines showed different ability in maturation. Over 80% cotyledonary somatic
embryos germinated normally on DCR medium containing 0.2% activated carbon. The success on SE induction of the species has
provided an effective clonal propagation method for this important tree’s genetic improvement. 相似文献
11.
Somatic embryos of Cyclamen persicum Mill. could be produced through a callus phase from juvenile explant material including anthers, ovaries and zygotic embryos. The auxin 2,4-D (1.0–1.5 mg l-1) and coconut milk (10% v/v) in MS medium were important factors for the induction of somatic embryogenesis. Somatic embryos germinated into plantlets in MS medium without growth regulators. The plants grew well in the greenhouse and flowered normally. The plants were phenotypically identical to the mother plants with a few exceptions.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
1-naphthylacetic acid
- IAA
3-indoleacetic acid
- BA
6-benzyladenine
- ABA
abscisic acid
- CM
coconut milk 相似文献
12.
Thomas L. Reynolds 《Plant cell reports》1986,5(4):273-275
The direct differentiation of bicellular pollen grains of Solanum carolinense L. (Horse-nettle; Solanaceae) into embryoids and plantlets was induced by culturing whole anthers on Murashige and Skoog's medium supplemented with IAA. The highest frequency of embryogenic induction occurred at 10 mg/l IAA. Developmentally, both the generative and vegetative cells of the pollen grain contributed to embryoid formation whose pattern of development was similar to that of zygotic embryos. In a previous study, it was show that 2,4-D promoted callus formation by pollen grains in cultured anthers of S. carolinense. It appears then that there are two distinct pathways of androgenesis in this species that are determined by the type of auxin present in the medium.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- IAA
indole-3-acetic acid
- BA
benzyladenine
- KIN
kinetin
- MS
Murashige and Skoog 相似文献
13.
Zygotic embryos at different developmental stages were tested for their potential in the initiation of embryogenic suspensor
mass (ESM) lines using immature seeds of Pinus rigida × P. taeda. The highest frequency (1.1%) of ESM was obtained with explants from cones collected on July 1. All excised embryos of the
July 1 collection were at the early proembryo stage. Two different culture media were compared. Forty-eight ESM lines were
initiated on Pinus taeda basal medium (P6) (0.97%) with 13.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.4 μM benzyladenine (BA). However, only
four ESM were obtained on a modified Murashige and Skoog medium (MSG; 0.55%). Most of the ESM arose from the seeds that were
at the stages ranging from late cleavage polyembryony to the early staged proembryo. Out of 52 lines (0.46%) that were produced
from 11,388 explants, only two viable lines (0.018%) (PRT11 and PRT28) survived. As for somatic embryo maturation, the highest
number (224/g−1 FW) of matured cotyledonary somatic embryos (line PRT 28) was obtained on a medium containing 100 μM abscisic acid (ABA),
0.2 M maltose, and 1.2% gellan gum. For germination of the somatic embryos, the cotyledonary somatic embryos derived from
maturation medium were transferred on half-strength Litvay medium (LM) plus 0.4% gellan gum. The germination rates were high
(71.4–96.3%) regardless of the concentrations of either ABA or gellan gum in the maturation medium. Approximately 500 somatic
plants were recovered from the germination medium and transferred to the green house; finally most of them were transplanted
successfully to the experimental field. 相似文献
14.
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
15.
Summary Media components used for three stages of development: (1) callus maintenance, (2) maturation of embryos, and (3) conversion
of embryos to plants were shown to affect regeneration of plants for the commercially important red rose cultivar Kardinal.
Embryogenic callus was maintained for 5yr on either Schenk and Hildebrandt’s basal salts medium (SH) supplemented with 13.6
μM 2,4-dichlorophenoxyacetic acid (2,4-D) or Murashige and Skoog’s basal salts medium (MS) supplemented with 18.1 μM dicamba and 0.46 μM kinetin. Maturation of embryos was three times higher using callus maintained on the SH medium supplemented with 2,4-D while
conversion of cotyledonary-stage embryos to plants was significantly higher (10 times) using callus that had been maintained
on MS medium with dicamba and kinetin. Maximum maturation (13.5%), and conversion (15.2%), occurred when callus was cultured
on MS maturation medium without hormones. Cotyledonary-stage embryos cultured on MS conversion medium supplemented with abscisic
acid (5–20 μM) produced plants that survived at a significantly higher rate (two times) in the greenhouse than when embryos were cultured
without abscisic acid. The highest rate of plant regeneration occurred when embryogenic callus of ‘Kardinal’ was maintained
on MS medium supplemented with dicamba and kinetin, maturation of embryos occurred on MS maturation medium without hormones,
and conversion of cotyledonary-stage embryos occurred on MS conversion medium supplemented with abscisic acid. 相似文献
16.
17.
Various tissues of seeds and seedlings of melon were cultured in vitro to study the effects of auxin concentration on organogenesis and embryogenesis. Adventitious shoots and somatic embryos were formed from explants of cotyledons of mature seeds, hypocotyls of seedlings, and leaves and petioles of young plantlets. Expanded cotyledons of seedlings formed only adventitious shoots. All tissues responded similarly to the 2,4-D concentration in the media, that is, adventitious shoots were formed at low concentration, callus proliferated without differentiation at intermediate concentration and somatic embryos were induced at high concentration. Cotyledons of mature seeds formed both adventitious shoots and somatic embryos more efficiently than any other tissues cultured.Effects of three auxins, 2,4-D, NAA and IAA, on organogenesis and embryogenesis were compared using cotyledons of mature seeds. Adventitious shoots were formed at low level of auxins (0 to 0.01 mg/l 2,4-D; 0 to 0.1 mg/l NAA; 0 to 1.0 mg/l IAA), and embryos were formed at high level of auxins (1.0 to 2.0 mg/l 2,4-D; 3.0 to 10.0 mg/l NAA; 20.0 to 100.0 mg/l IAA). IAA gave more efficient shoot formation and embryogenesis than the other auxins.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- IAA
3indoleacetic acid
- BA
6-benzylaminopurine
- MS
Murashige and Skoog 相似文献
18.
A method for the induction of somatic embryos in soybean tissue cultures is described. Cotyledons from immature embryos were utilized as explant source. Supplementing the culture medium with auxins (2,4-D, MCPA, 2,4,5-T, NAA, IAA, IBA) caused formation of meristematic tissue on cotyledon explants. The extent of meristematic tissue formed depended on the kind and concentration of auxin in the culture medium. With 2,4-D and MCPA, embryoids originated from meristematic tissue. Embryoid formation rates were influenced by the developmental stage of the embryos serving as explant source and auxin concentration. Addition of cytokinins to the medium containing 2,4-D or supplementing it with high sugar concentrations inhibited the formation of meristematic tissue and of embryoids on cotyledon explants.Abbreviations BA
6-benzyladenin
- 2,4-D
2,4-dichlorphenoxyacetic acid
- IAA
indoleacetic acid
- IBA
indolebutyric acid
- MCPA
2-methyl-4-chlorophenoxyacetic acid
- NAA
-naphthaleneacetic acid
- 2,4,5-T
2,4,5-trichlorophenoxyacetic acid
- L2
Phillips and Collins (1979) medium
Present and correspondence address: Akademie der Landwirtschaftswissenschaften der DDR, Institut für Pflanzenernährung, DDR-6909, Jena 相似文献
19.
Liberato Portillo Fernando Santacruz-Ruvalcaba Antonia Gutiérrez-Mora Benjamín Rodríguez-Garay 《In vitro cellular & developmental biology. Plant》2007,43(6):569-575
Somatic embryogenesis was achieved from leaves of Agave tequilana Weber cultivar azul utilizing MS medium supplemented with L2 vitamins and the addition of cytokinins: 6-benzylaminopurine
(BA), 1-phenyl-3(1,2,3-thiadiazol-5-yl)urea (TDZ), 6-(γ-γ-dimethylamino)purine (2ip) and 6-furfurylaminopurine (KIN), combined
with the auxin 2,4-dichlorophenoxyacetic acid (2,4-D). Differences among the six genotypes studied with regard to their embryogenic
response in culture were found. Embryos produced by genotype S3 under a hormone regime of high cytokinin (44.4 to 66.6 μM
BA) compared to auxin (4.5 μM 2,4-D) contained chlorophyll, whereas those produced when auxin was high compared to cytokinin
(9.0 and 13.6 μM 2,4-D and 1.3 and 4.0 μM BA, respectively) were whitish and morphologically similar to their zygotic counterparts.
Somatic embryos matured and germinated after transferring the embryogenic calli to maturation and germination medium without
growth regulators and enriched with organic nitrogen. Microscopic observations demonstrated a unicellular origin for production
of indirect somatic embryos. 相似文献
20.
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