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1.
Jun Ma Yehua He Chenghou Wu Heping Liu Zhongyi Hu Guangming Sun 《Plant Molecular Biology Reporter》2012,30(1):195-203
A somatic embryogenesis receptor-like kinase (SERK) gene, designated as AcSERK1, was isolated from pineapple (Ananas comosus cv. Shenwan). AcSERK1 shared all the characteristic domains of the SERK family, including five leucine-rich repeats, one proline-rich region motif,
transmembrane domain, and kinase domains. Somatic embryogenic cultures of pineapple were established following transfer of
callus cultures to Murashige and Skoog (1962) medium containing 2,4-dichlorophenoxyacetic acid. The role of AcSERK1 during establishment of somatic embryogenesis in culture was investigated. The AcSERK1 was highly expressed during embryogenic competence acquisition and global embryo formation in culture. These findings were
obtained along with morphological changes in callus cultures exhibiting embryogenic potential. Overall, levels of expression
of AcSERK1 were lower in nonembryogenic tissues and organs than in embryogenic callus. In situ hybridization analysis revealed that
AcSERK1 expression was detected in embryogenic tissues, including single competent cells, meristematic centers wherein embryogenic
structures are formed, and global embryos. These results suggested that AcSERK1 expression was associated with induction of somatic embryogenesis and that it could be used as a potential marker gene to
monitor the transition of pineapple callus tissues into competent and embryogenic cells and tissues. 相似文献
2.
M. C. San-José E. Corredoira M. T. Martínez N. Vidal S. Valladares R. Mallón A. M. Vieitez 《Plant cell reports》2010,29(6):661-671
A procedure for inducing somatic embryos in shoot apex explants (2 mm) excised from shoot proliferation cultures established
from adult oak trees (Quercus robur) was investigated. Embryogenesis was induced in shoot tip as well as leaf explants in three out of the five genotypes evaluated.
Somatic embryos were formed by culture in induction medium supplemented with 21.48 μM naphthalene acetic acid and 2.22 μM
benzyladenine for 8 weeks, and successive transfer of explants to expression media with a low concentration of growth regulators
and without them. Both types of explants formed callus tissue from which somatic embryos developed, indicating indirect embryogenesis.
Although the embryogenic frequencies were lower than 12%, it did not prevent the establishment of clonal embryogenic lines
maintained by repetitive embryogenesis. Histological study confirmed an indirect somatic embryogenesis process from shoot
tip explants, in which leaf primordia and the corresponding axial zones were involved in generating callus, whereas the apical
meristem itself did not proliferate. The origin of embryogenic cells appeared to be associated with dedifferentiation of certain
parenchymal cells in callus regions after transfer of explants to expression media without auxin. Division of embryogenic
cells gave rise to proembryo aggregates of unicellular origin, although a multicellular origin from bulging embryogenic areas
would also seem possible. Further development led to the formation of cotyledonary-stage somatic embryos and nodular embryogenic
structures that may be considered as anomalous embryos with no clear bipolarity. Inducement of somatic embryos from explants
isolated from shoot cultures ensures plant material all year round, thus providing a significant advantage over the use of
leaf explants from field-grown trees. 相似文献
3.
4.
Heung Kyu Moon So Young Park Yong Wook Kim Sea Hyun Kim 《In vitro cellular & developmental biology. Plant》2008,44(2):119-127
An effective micropropagation technique via somatic embryogenesis has been developed using tissue from serially grafted shoots
generated from a mature Kalopanax septemlobus tree (~40 y old). Callus was induced from leaf segments obtained from the grafts by culturing the explants in Murashige and
Skoog (MS) medium supplemented with 2,4-D and 3% w/v sucrose under darkness. The effects of sucrose, coconut water, and polyethylene glycol (PEG-3350) were evaluated as factors
to promote development of somatic embryos (SEs) from embryogenic callus. More than 90% of explants formed callus; however,
only 2.5%, or 20 leaf segments out of 800 explants, formed embryogenic callus after 8 wk of culture. High sucrose concentrations
(3% and 5% w/v) were effective in inducing SEs. Treatment with 2–10% v/v coconut water also had a positive effect on embryo induction. A synergistic effect on SE induction was obtained using sucrose
and PEG, with presence of the latter compound resulting in smaller, more uniform SEs. Embryo germination and conversion to
plantlets were significantly influenced by the gelling agents. In general, gelrite-gelled medium was superior to agar-gelled
medium. In gelrite-gelled medium, gibberillic acid (GA3) enhanced embryo germination. Converted plantlets in an artificial soil mixture showed a 91% survival rate and displayed
no distinct morphological variations. Our results indicate that reliable somatic embryogenesis and plant production can be
achieved with rejuvenated tissues after repeated grafting of shoots derived from a mature Kalopanax septemlobus tree. 相似文献
5.
Background and Aims
Understanding the fate and dynamics of cells during callus formation is essential to understanding totipotency and the mechanisms of somatic embryogenesis. Here, the fate of leaf explant cells during the development of embryogenic callus was investigated in the model legume Medicago truncatula.Methods
Callus development was examined from cultured leaf explants of the highly regenerable genotype Jemalong 2HA (2HA) and from mesophyll protoplasts of 2HA and wild-type Jemalong. Callus development was studied by histology, manipulation of the culture system, detection of early production of reactive oxygen species and visualization of SERK1 (SOMATIC EMBRYO RECEPTOR KINASE1) gene expression.Key Results
Callus formation in leaf explants initiates at the cut surface and within veins of the explant. The ontogeny of callus development is dominated by the division and differentiation of cells derived from pluripotent procambial cells and from dedifferentiated mesophyll cells. Procambium-derived cells differentiated into vascular tissue and rarely formed somatic embryos, whereas dedifferentiated mesophyll cells were competent to form somatic embryos. Interestingly, explants incubated adaxial-side down had substantially less cell proliferation associated with veins yet produced similar numbers of somatic embryos to explants incubated abaxial-side down. Somatic embryos mostly formed on the explant surface originally in contact with the medium, while in protoplast microcalli, somatic embryos only fully developed once at the surface of the callus. Mesophyll protoplasts of 2HA formed embryogenic callus while Jemalong mesophyll protoplasts produced callus rich in vasculature.Conclusions
The ontogeny of embryogenic callus in M. truncatula relates to explant orientation and is driven by the dynamics of pluripotent procambial cells, which proliferate and differentiate into vasculature. The ontogeny is also related to de-differentiated mesophyll cells that acquire totipotency and form the majority of embryos. This contrasts with other species where totipotent embryo-forming initials mostly originate from procambial cells.Key words: Callus, dedifferentiation, leaf veins, Medicago truncatula, pluripotency, procambium, protoplasts, reactive oxygen species, SERK, somatic embryogenesis, stem cells, totipotency 相似文献6.
Embryogenic cells in Dactylis glomerata L. (Poaceae) explants identified by cell tracking and by SERK expression 总被引:6,自引:0,他引:6
Single mesophyll cells in leaf explants of Dactylis glomerata L. (Dactylis) that were competent to form somatic embryos directly or through callus were identified by semi-automatic cell tracking.
These competent cells were a subpopulation of small, isodiametric, cytoplasm-rich cells located close to the vascular bundles.
Using whole mount in situ hybridization, we showed that a similar subpopulation of cells expressed the Somatic Embryogenesis
Receptor-like Kinase (SERK) gene during the induction of embryogenic cell formation. In both leaf explants and suspension
cultures, a transient pattern of SERK gene expression was found during early embryo development, up to the globular stage.
In later embryo stages, SERK mRNA was present in the shoot apical meristem, scutellum, coleoptile and coleorhiza.
Received: 14 May 1999 / Revision received: 27 August 1999 / Accepted: 8 September 1999 相似文献
7.
Jos Van Boxtel Albertus Eskes Marc Berthouly 《In vitro cellular & developmental biology. Plant》1997,33(1):6-12
Summary To improve selection of transgenic Coffea spp. tissue after transformation treatments, the effects of the selective agents chlorsulfuron, glufosinate, glyphosate,
hygromycin, and kanamycin were studied on callus development from leaf explants (from greenhouse-grown plants and somaplants)
and in embryogenic suspension cultures. Studied genotypes were from C. arabica, C. canephora, and the interspecific hybrids Arabusta and Congusta. A culture system based on “direct” somatic embryogenesis from C. canephora leaf explants proved to be more sensitive to selective agents than high frequency somatic embryogenesis from C. arabica or Arabusta leaf explants. With respect to the selective effect, chlorsulfuron and hygromycin provoked strong inhibition
and severe necrosis, whereas glyphosate and kanamycin showed variable inhibition. Glufosinate appeared to efficiently inhibit
growth of both leaf callus and callus suspensions of all genotypes tested without inducing necrosis. These properties may
make the use of glufosinate advantageous in a selective growth system for detection of transformed coffee tissues. 相似文献
8.
Xin Hu Chun-Rong Zhang Hui Xie Xia Huang Yun-Feng Chen Xue-Lin Huang 《Acta Physiologiae Plantarum》2012,34(3):1067-1074
Homeodomain leucine zipper (HD-Zip) proteins play important roles in plant development. In this study, we not only identified
and characterized a new HD-Zip II gene, designated as MSHB1 (HM114227), from alfalfa (Medicago sativa L. cv. Jinnan) callus treated with thidiazuron (TDZ) which reduced the embryogenic competence of the callus, but also presented
the first evidence that MSHB1 is involved in the inhibitory effect of TDZ on somatic embryogenic competence in alfalfa callus. The full-length cDNA was
1,578 bp with an open reading frame of 1,023 bp, encoding a predicted protein of 340 amino acid residues, plus three introns.
MSHB1 was strongly expressed in the callus treated with TDZ, but was only slightly detected in the leaf and petiole. TDZ treatment
significantly decreased the frequency of somatic embryogenesis in the callus, but up-regulated MSHB1 expression during callus induction, callus maintenance and somatic embryo induction. These results suggest that the inhibitory
effect of TDZ on embryogenic competence of alfalfa callus might be mediated by the regulation of MSHB1 expression. 相似文献
9.
Thiago E. R. Silva Luciana C. Cidade Fátima C. Alvim Júlio C. M. Cascardo Marcio G. C. Costa 《Plant Cell, Tissue and Organ Culture》2009,99(3):287-298
In order to develop a more efficient genetic transformation system for cacao somatic embryos, the effects of polyamines and
β-lactam antibiotics on somatic embryogenesis, hygromycin as selective agent, and different factors affecting uidA gene transfer have been evaluated. The polyamines putrescine, spermidine, and spermine significantly improved secondary somatic
embryogenesis in cacao. Spermine at 1,000 μM provided the best responses, increasing 6.7× the percentage of embryogenic callus
and 2.5× the average number of embryos per embryogenic callus. The β-lactam antibiotics timentin and meropenem, used for Agrobacterium tumefaciens counter-selection, had a non-detrimental effect on secondary somatic embryogenesis, depending on their concentration, whereas
the commonly used β-lactam cefotaxime inhibited it, irrespective of the tested concentration. Hygromycin showed a strong inhibitory
effect on secondary somatic embryogenesis of cacao, impairing completely the embryo production at 20 mg l−1. Following the criterion of GUS activity, the best conditions for T-DNA transfer into cotyledon explants from primary somatic
embryos of cacao were a sonication of the explants for 100 s, a 20-min incubation period in Agrobacterium solution, an Agrobacterium concentration of 1.0 (OD600), and cocultivation of the explants on tobacco feeder layers. These findings will have important implications for studies
on functional genomics of cacao. 相似文献
10.
A simple and efficient system was developed for rapid somatic embryogenesis from leaf explants of Merwilla plumbea, a traditional but threatened medicinal plant in South Africa. Friable embryogenic callus (FEC) was obtained from leaf explants
on embryogenic callus induction medium containing agar-solidified Murashige and Skoog (MS) salts and vitamins, 8.3 μM picloram,
2.3 μM thidiazuron (TDZ) and 20 μM glutamine. FEC was subsequently incubated in embryogenic callus proliferation medium containing
4.5 μM 2,4-dichlorophenoxyacetic acid (2,4-D) and 4.1 μM picloram for 7 days before it was transferred to liquid somatic embryo
medium (SEML) containing MS medium supplemented with 0.4 μM picloram and 0.9 μM TDZ. In SEML supplemented with 150 mg L−1 haemoglobin, 5.4–35.6 somatic embryos per settled cell volume of 500 mg FEC were obtained. These embryos were at globular
to cotyledonary developmental stages. Embryo maturation, germination and plant formation rate was 94.4% following transfer
of SEs to half-strength MS medium supplemented with 1.4 μM gibberellic acid. Plantlets transferred into soil acclimatized
in the misthouse and established successfully in the greenhouse (100%). This is the first report on induction of Merwilla plumbea somatic embryogenesis. The protocol developed offers controlled vegetative propagation by alleviating extinction threats,
ensures germplasm conservation and provides a system for physiological, biochemical, molecular and cellular studies of embryo
development. 相似文献
11.
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
12.
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
13.
In vitro somatic embryogenesis in two major rattan species: Calamus merrillii and Calamus subinermis
D. K. S. Goh M. -C. Bon F. Aliotti J. Escoute N. Ferrière O. Monteuuis 《In vitro cellular & developmental biology. Plant》2001,37(3):375-381
Summary Occurrence of somatic embryogenesis in in vitro cultures of Calamus merrillii and Calamus subinermis, two major largecaned rattan species, was scientifically demonstrated for the first time. Tissue responsiveness varied markedly
according to the species and the type of primary explants used when initiated on 10.4–31.2 μM picloram-enriched Murashige and Skoog callus induction media. In C. merrillii, within 6 wk after inoculation, 84% of the leaf and 90% of the zygotic embryo explants produced friable embryogenic calluses,
by contrast with those formed by 74% of the root explants. In C. subinermis, callogenesis was observed only 6 mo. after inoculation in 68% of root and 48% of zygotic explants. Leaf explants did not
respond at all. Only root-derived calluses developed into nodular embryogenic structures. Irrespective of these initial differences,
the further steps of the somatic embryogenesis developmental pattern was similar for both species. Histological analyses established
that callus formation took place in the perivascular zones, and could give rise to embryogenic isolated cells from which the
proembryos were derived. Reducing the picloram concentration stimulated the maturation process resulting ultimately in the
germination of somatic embryos that exhibited bipolar development, despite an apparent lack of starch and protein reserves.
The somatic embryo-derived plantlets of C. merrillii, overall more prone to somatic embryogenesis than C. subinermis in the given conditions, were successfully acclimatized to outdoor conditions. 相似文献
14.
The induction of somatic embryogenesis from shoot apices and leaf explants of shoot cultures derived from 6- to 7-year-old
white oak (Quercus alba L.) trees is reported in this study. Embryogenic response was obtained in two out of the three genotypes evaluated with embryo
induction frequencies up to 50.7% for WOQ-1 and 3.4% for WOQ-5 genotypes. The embryogenic explants formed translucent nodular
structures and cotyledonary-stage somatic embryos, which developed from callus tissue, indicating an indirect embryogenesis
process. An efficient procedure was developed for WOQ-1 material on the basis of the most appropriate leaf developmental stage.
Growing leaves excised from two nodes below the shoot apex showed the highest embryogenic induction index. These leaves contain
cells in an undifferentiated state, as shown by the presence of precursor cells of stomata, absence of intercellular spaces
and low starch content in the mesophyll cells. Nodular structures and/or somatic embryos began to arise 7–8 weeks after culture
initiation, although most emerged after 9–12 weeks in culture. The sequence of application of media for somatic embryo induction
was optimized with a two-step procedure consisting of culturing the explants in medium supplemented with 21.48 μM NAA and
2.22 μM BA for 8 weeks and transfer of explants into plant growth regulator-free medium for another 12 weeks. Clonal embryogenic
lines were established and maintained by secondary embryogenesis. Embryo germination (30%) and plantlet conversion (16.6%)
were achieved after cold storage for 2 months. 相似文献
15.
B. Cuenca M. C. San-José M. T. Martínez A. Ballester A. M. Vieitez 《Plant cell reports》1999,18(7-8):538-543
Internodal and leaf segments from pedunculate oak (Quercus robur L.) seedlings were used as explant source to induce somatic embryogenesis. Auxin treatment influenced embryogenic response, which
only occurred in explants initially cultured on media containing 4 mg/l naphthaleneacetic acid (NAA) and different benzyladenine
(BA) concentrations. After 6 weeks of culture on induction medium, the explants were transferred to medium supplemented with
0.1 mg/l BA and 0.1 mg/l NAA, and 4 weeks later, they were subcultured in a growth-regulator-free medium, in which somatic
embryos arose through indirect regeneration on the surface of a nodular callus. Somatic embryos were induced in explants of
two out of four seedling provenances. The induction frequency ranged from 16% in leaf explants to 4% in internodal explants.
Somatic embryos developed two cotyledons, which were translucent or opaque-white in appearance, but anomalous morphologies
were also observed. Different embryogenic lines were established and maintained by repetitive embryogenesis in multiplication
medium containing 0.1 mg/l BA plus 0.05 mg/l NAA. These results indicate that tissues from explants other than Q. robur zygotic embryos are able to produce embryogenic cultures.
Received: 14 July 1998 / Revision received: 2 November 1998 / Accepted: 6 November 1998 相似文献
16.
Ananya Paul Kalyan Mitter Sarmistha Sen Raychaudhuri 《Plant Cell, Tissue and Organ Culture》2009,97(3):303-311
The effects of exogenous polyamines (PAs) on enhancement of somatic embryogenic calli was investigated in Momordica
charantia L. in vitro. Induction of somatic embryogenesis (SE) in leaf explants of M. charantia after 21 days of culture in Murashige and Skoog (MS) medium was determined using scanning electron microscopy. During induction
of SE there were high titers of Putrescine (Put) as compared to Spermidine (Spd) and Spermine (Spm), a prerequisite for cell
division. Addition of PAs to the embryogenic media resulted in an increase in fresh weights and number of somatic embryos
of 21-day old embryogenic calli. Put at a concentration of 1 mM showed maximum increase in fresh weights of embryogenic calli
(5 fold) and number of somatic embryos produced per 0.2 g of callus (2.5 fold). Moreover addition of PAs to the embryogenic
media resulted in lowering of endogenous free PA level of 21-day old embryogenic calli. Thus, when the media was supplemented
with exogenous PAs a positive correlation was found to exist between Somatic Embryogenesis enhancement and decrease in endogenous
free PA levels. 相似文献
17.
Tina Lopes Ana Capelo Gina Brito João Loureiro Conceição Santos 《Trees - Structure and Function》2009,23(1):29-36
The crop species Olea europaea L. (olive tree) is of great economic importance in the Mediterranean region. Hence, many efforts have been done in the last
decades to propagate this commercially valuable species by in vitro methods. On the other hand, the lesser known Olea maderensis (Lowe) Rivas Mart. & Del Arco which is a native species of the Madeira Archipelago has only been the subject of micropropagation
from nodal stem cuttings. Therefore, in this work we analysed the stability of ten nuclear simple sequence repeat (SSR) markers
at successive stages of the somatic embryogenesis process in two adult trees belonging to these two species from the Madeira
Archipelago. For the induction of somatic embryogenesis, petiole and leaf explants were cultivated on solid Murashige and
Skoog medium (MS) with 12.25 μM indole-3-butyric acid (IBA) and 4.56 μM of zeatin, in the dark. After 3 months, different
callus tissues (non-embryogenic, pre-embryogenic and embryogenic) developed from leaf explants and petioles were later transferred
to MS medium without growth regulators in the dark. All ten SSR markers were able to distinguish between species. However,
no mutations were found at the SSR loci at any of the successive developmental stages from PEMs (pre-embryogenic masses) to
somatic embryos. This genetic uniformity was observed within material derived from each genotype/species and its respective
donor plant. Therefore, we conclude that the genomic integrities of both O. europaea and O. maderensis were maintained throughout the stages of the embryogenic processes in study suggesting the absence of somaclonal variation. 相似文献
18.
The halophyte Leymus chinensis (Trin.) is a perennial rhizome grass (tribe Gramineae) that is widely distributed in China, Mongolia and Siberia, where it
is produced as a forage product. In this report, we establish a highly reproducible plant regeneration system through somatic
embryogenesis. Two explants, mature seeds and leaf base segments were used; these parts displayed different responses to combinations
of growth factors that affect embryogenic callus induction, callus type optimization and plant regeneration. The highest callus
induction frequency was obtained on Murashige and Skoog (MS) medium supplemented with 2.0 mg l−1 2,4-dichlorophenoxyacetic acid (2,4-D) in the presence of 5.0 mg l−1
l-glutamic acid. The inclusion of 5.0 mg l−1
l-glutamic acid was found to significantly promote primary callus induction, embryogenic callus formation and callus status
improvement. Subculturing on maintenance medium for 1–2 months before plant regeneration was found to be essential for the
optimization of callus type and the maturation of embryogenic callus. Callus relative water content and growth rate were simultaneously
investigated during callus maintenance, and found to possibly be related to callus type. Shoots were differentiated from the
embryogenic callus on the optimal medium with MS salts containing 0.2–0.5 mg l−1 α-naphthalene acetic acid (NAA), 2.0 mg l−1 kinetin (Kn) and 2.0 g l−1 casamino acids in 71.0 and 69.2% of wild-type (WT) and Jisheng No.1 (JS) plants, respectively. Plant regeneration was variable
depending on NAA levels, and the addition of casamino acids stimulated the maturation of embryogenic callus and plant regeneration.
Transferring callus with shoots onto half-strength MS medium resulted in rooting within 1 week. The growth of regenerated
plants was also surveyed in the field. This is the first report of plant regeneration through somatic embryogenesis from mature
seeds and leaf base segments of L. chinensis. 相似文献
19.
W. K. Ho S. E. Ooi S. Mayes P. Namasivayam M. Ong-Abdullah C. F. Chin 《Tree Genetics & Genomes》2013,9(4):1099-1107
The association between DNA methylation status and embryogenic competency in oil palm tissue culture was examined through Representational Difference Analysis (RDA) approach, using methylation-sensitive restriction endonucleases. “Difference Products” (DPs) of RDA derived from palms of similar genetic backgrounds but exhibiting different embryogenesis rates during the regeneration process were isolated. The DPs were sequenced using a pyrosequencing platform. To our knowledge, this is the first study profiling partial HpaII methylation sites in oil palm young leaf tissues which are potentially associated with embryogenic amenability through a genomic subtractive approach. Quantitative real-time PCR analysis demonstrated that the methylation status of a novel fragment, EgNB3, was higher in highly embryogenic leaf explants compared to low embryogenesis rate materials. These differences are likely to be contributed by the 5′-mCCGG-3′ and/or 5′-mCmCGG-3′ methylation patterns. Our data suggest that the differentially methylated site in EgNB3 has potential as a molecular biomarker for the screening of oil palm leaf explants for their embryogenic potentials. 相似文献
20.
M. Muruganantham S. Amutha A. Ganapathi 《In vitro cellular & developmental biology. Plant》2010,46(1):34-40
The regeneration of plants via somatic embryogenesis liquid shake culture of embryogenic calluses was achieved in Vigna mungo (L.) Hepper (blackgram). The production of embryogenic callus was induced by seeding primary leaf explants of V. mungo onto Murashige and Skoog (MS) (Physiol Plant 15:473–497, 1962) medium supplemented (optimally) with 1.5 mg/l 2,4-dichloro-phenoxyacetic acid. The embryogenic callus was then transferred
to liquid MS medium supplemented (optimally) with 0.25 mg/l 2,4-dichloro-phenoxyacetic acid. Globular, heart-shaped, and torpedo-shaped
embryos developed in liquid culture. The optimal carbohydrate source for production of somatic embryos was 3% sucrose (compared
to glucose, fructose, and maltose). l-Glutamine (20 mg/l) stimulated the production of all somatic embryo stages significantly. Torpedo-shaped embryos were transferred
to MS (Physiol Plant 15:473–497, 1962) liquid medium containing 0.5 mg/l abscisic acid to induce the maturation of cotyledonary-stage embryos. Cotyledonary-stage
embryos were transferred to 1/2-MS semi-solid basal medium for embryo conversion. Approximately 1–1.5% of the embryos developed
into plants. 相似文献