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1.
The effects of glutamine-based dipeptides, glutamine and casein hydrolysate, as well as the deletion of organic nitrogen, were investigated during white spruce [Picea glauca (Moench) Voss] somatic embryogenesis. There were no differences in the fresh weight increase of the tissue masses grown on initiation medium with different combinations of organic nitrogen. This was also the case for subsequent growth on kinetin medium, except that glutamine alone produced a significantly lower fresh weight increase than the other organic nitrogen combinations. Without organic (i.e. with only inorganic) nitrogen in the medium, the fresh weight increase was significantly less than with organic nitrogen on both initiation and kinetin medium. No differences were found between the dry/fresh weight ratios obtained with the various nitrogen treatments. The number of mature embryos produced per gram fresh weight when cultured in the absence of organic nitrogen was significantly higher than that obtained in its presence. There were no differences in the total number of mature embryos produced in cultures grown with various organic nitrogen combinations or without organic nitrogen. There were large clone differences with respect to the number of mature somatic embryos per gram tissue and the total number of somatic embryos produced. Hence, nitrogen type influences culture growth rate but not the number of mature somatic embryos produced. The latter was clone dependent.Abbreviations
BA
6-benzylaminopurine
-
CH
casein hydrolysate 相似文献
2.
B. Helliot D. Madur E. Dirlewanger M. T. De Boucaud 《In vitro cellular & developmental biology. Plant》2002,38(5):493-500
Summary The evaluation of the genetic stability of Prunus Ferlenain plants regenerated from cryopreserved apices was investigated. The analysis of plants recovered from frozen material
was performed at the phenotypic (developmental competence), cytological (chromosomal preparations) and molecular level [random
amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP)]. No genetic change was detected among
the plantlets regenerated from frozen apices in comparison to the non-frozen material, including leaves of the mother tree
kept in an orchard and vitroplantlets regenerated from non-frozen apices. This result suggested that the procedure used for
Prunus cryopreservation could be used for long-term conservation. The relevance of each strategy for the genetic stability evaluation
of the cryopreserved material is discussed. 相似文献
3.
4.
Summary The glutathione-glutathione disulfide redox pair was utilized to improve white spurce somatic embryo development. Mature cotyledonary-stage
somatic embryos were divided into two groups (A and B) based on morphological normality and the ability of the mature somatic
embryos to convert into plantlets. Group A embryos had four or more cotyledons and converted readily upon germination after
a partial drying treatment. Group B embryos had three or fewer cotyledons with a low conversion frequency. The addition of
reduced glutathione (GSH) at a concentration of 0.1 mM resulted in an increase in embryo production (total population) with a mean total number of 64 embryos per 100 mg embryogenic
tissue as well as an increase in post-embryonic root growth. However, at a higher concentration (1 mM), GSH inhibited embryo formation. The manipulation of the tissue culture environment via the inclusion of glutathione disulfide
(GSSG), at concentrations of 0.1 and 1.0 mM, enhanced the development of better-quality embryos. This quality was best exemplified when embryos forming four or more
cotyledons increased by at least twofold to 73.9% when treated with 1.0 mM GSSG, compared to 38% in control. Furthermore, this improved quality was reflected by an increased conversion frequency.
A 20% increase in the ability of the somatic embryo to produce both root and shoot structures during post-embryonic development
was noted when embryos were matured on maturation medium supplemented with 1.0 mM GSSG over the control. 相似文献
5.
Claudio Stasolla Edward C. Yeung 《In vitro cellular & developmental biology. Plant》1999,35(4):316-319
Summary The effects of exogenous applications of ascorbic acid on white spruce somatic embryogenesis were examined. Increasing concentrations
of ascorbate (1 μM to 100 μM) in the germination medium enhanced somatic embryo conversion in a linear fashion. At the optimal ascorbate level (100 μM) the number of embryos able to undergo normal conversion, i.e., emergence of both root and shoot, increased from 34% (control)
to 58%. The effect of ascorbate had a more pronounced effect on shoot growth than on root emergence; and at 100 μM ascorbate, the percentage of embryos able to produce new leaf primordia increased from 47% (control) to 79%. Root emergence
increased slightly from 64% in the control embryos to 74% in the presence of ascorbic acid. The ascorbate-treated embryos
were characterized by an enlarged apical region, presumably due to a larger number of leaf primordia produced, and by dark
green leaves. When allowed to grow further, these embryos were able to develop into normal plantlets. 相似文献
6.
Simon D. Moorhouse Graham Wilson Michael J. Hennerty Christopher Selby Seán Mac An tSaoir 《Plant Growth Regulation》1996,20(1):53-56
The Braun Biostat BF2 bioreactor system employs a novel aeration and agitation system, designed to enhance gaseous exchange and reduce shear stresses on submerged cell suspension cultures. The Biostat BF2 bioreactor employs a central pivoting spindle, around which the aeration tubing is wound forming a large paddle-type structure suspended from the top-plate and swung in a circle by a solid-state magnetic stirrer.The aeration tubing is a polypropylene capillary membrane, which has a unique microporous structure and is ideal for aeration, permitting two-way, bubble-free, gaseous exchange of the medium. This tubing can be rendered porous and can be used in the perfusion of aqueous solutions, enabling cell-free media exchange to be conducted. Thin-walled silicone rubber tubing, although gas permeable to a degree, cannot be made porous to aqueous solutions.The bioreactor was inoculated with a suspension culture of Sitka spruce (Picea sitchensis [Bong.] Carr.) known to be embryogenic and capable of maturing to plantlets on solidified medium. The perfusion capability of the bioreactor was employed to replace the inital proliferation medium with maturation medium in order to induce the development of the somatic embryos in submerged cell culture. The size ratio of the somatic embryo heads was monitored over 7 weeks. This cell line was found to mirror just the initial elongation, previously observed in shake-flask culture.Abbreviations BAP
6-benzylaminopurine
- 2,4-D
2,4-dichlorophenoxyacetic acid
- SSPM
Selby Sitka proliferation medium
- SSMM
Selby Sitka maturation medium
The following was presented at the NERC TBLG '95 Meeting as the Bioreactor Workshop 相似文献
7.
Norway spruce somatic embryogenesis: high-frequency initiation from light-cultured mature embryos 总被引:8,自引:0,他引:8
Somatic embryos and rooted plantlets have been regenerated from light-initiated embryogenic callus derived from mature embryos of Picea abies. Under a 16 h photoperiod, mature zygotic embryos were cultured on a modified half-strength Murashige & Skoog medium without NH4NO3 and supplemented with 5 mM glutamine, 4.5 M N6-benzyladenine and 10.7 M naphthaleneacetic acid or 10 M 2,4-dichlorophenoxyacetic acid. White translucent embryogenic callus, proliferating from the callusing hypocotyl region after 3 weeks incubation, was isolated from the green non-embryogenic tissue and subcultured for over 12 months. Upon transfer of the embryogenic callus through a specific sequence of media, somatic embryos proceeded to mature, elongating and forming rings of cotyledonary leaves similar to those of zygotic embryos. Transferred to medium without growth regulators, the somatic embryos germinated and produced plantlets with green cotyledons, elongated hypocotyls and primary roots. 相似文献
8.
Embryogenic cultures of pumpkin (Cucurbita pepo L.) were initiated from mechanically wounded mature zygotic embryos on 2,4-D-containing MS medium, and on hormone-free, semisolid modified MS medium containing NH4Cl as the sole source of nitrogen. The habituated line was derived from the embryogenic tissue induced with 2,4-D and maintained on medium without growth regulators. Sustained subculturing of the three embryogenic lines on a medium with NH4Cl as the sole source of nitrogen enabled the establishment of highly uniform cultures in which no further development into mature embryo stages occurred. The tissue consisting of proembryogenic globules or globular stage embryos was maintained, without decline, for over six years. Globular embryos proceeded to maturity when a combination of reduced (NH4) and unreduced (NO3) forms of nitrogen was provided in the medium. Different nitrogen sources in the medium caused changes of medium pH during subculture in the pH range of 4.0-6.5. The tissue growth and embryo development were blocked on medium with pH adjusted and stabilized at 4.0 or at 3.2. 相似文献
9.
Plant regeneration via somatic embryogenesis in cotton 总被引:6,自引:0,他引:6
An efficient in vitro plant regeneration system characterized by rapid and continuous production of somatic embryos using leaf and stem explants
of abnormal seedling as an explant have been developed in Gossypium hirsutum L. Embryogenic callus and somatic embryos have been obtained directly from the explants of cotton abnormal seedlings. Plant
growth regulators influenced the induction of cotton somatic embryogenesis. The optimal medium for direct somatic embryogenesis
was modified MS medium supplemented with 0.1 mg l-1 ZT and 2 g l-1 activated carbon. On this medium, an average of 28.0 and 28.1 matured somatic embryos formed from per leaf and stem explants
respectively. The highest frequency of somatic embryogenesis was 100%. The somatic embryos were converted into normal plantlets
when cultured on modified MS medium supplemented with 0.1 mg l-1 ZT. Upon transfer to soil, plants grew well and appeared normal. Plants could be regenerated within 60–80 days. The system
of cotton somatic embryogenesis and plant regeneration described here will facilitate the application of plant tissue culture
and genetic engineering on cotton genetic improvement.
This revised version was published online in June 2006 with corrections to the Cover Date. 相似文献
10.
Ravindra B. Malabadi K. Nataraja 《In vitro cellular & developmental biology. Plant》2006,42(2):152-159
Summary The present investigation reports optimized parameters for somatic embryogenesis and cryopreservation of embryogenic cultures
using shoot apical domes from mature trees of Pinus roxburghii Sarg. Embryogenic tissue of P. roxburghii Sarg. was cryopreserved for 24 h, 10 d, and 8 wk using sorbitol and dimethylsulfoxide (DMSO) as cryoprotectants. Results
indicate that 0.2M sorbitol and 5% DMSO had the best cryoprotecting effect. The recovered tissue showed luxuriant growth on maintenance medium
(II). Partial desiccation of thawed embryogenic tissue for 24 h prior to transfer to maturation medium enhanced the maturation
of somatic embryos. Maturation frequency increased from 1.3 to 18.3% after 12 h desiccation treatment, and from 18.3 to 61.8%
after 24 h of desiccation. However, non-desiccated embryogenic tissue produced the least number of somatic embryos (1.3%)
on the maturation medium with the same abscisic acid and Gellan gum concentration. All the three embryogenic lines produced
plantlets and had the same appearance and normal growth as compared to unfrozen controls. 相似文献
11.
Analysis of genetic stability at SSR loci during somatic embryogenesis in maritime pine (Pinus pinaster) 总被引:1,自引:0,他引:1
Liliana Marum Margarida Rocheta João Maroco M. Margarida Oliveira Célia Miguel 《Plant cell reports》2009,28(4):673-682
Somatic embryogenesis (SE) is a propagation tool of particular interest for accelerating the deployment of new high-performance
planting stock in multivarietal forestry. However, genetic conformity in in vitro propagated plants should be assessed as
early as possible, especially in long-living trees such as conifers. The main objective of this work was to study such conformity
based on genetic stability at simple sequence repeat (SSR) loci during somatic embryogenesis in maritime pine (Pinus pinaster Ait.). Embryogenic cell lines (ECLs) subjected to tissue proliferation during 6, 14 or 22 months, as well as emblings regenerated
from several ECLs, were analyzed. Genetic variation at seven SSR loci was detected in ECLs under proliferation conditions
for all time points, and in 5 out of 52 emblings recovered from somatic embryos. Three of these five emblings showed an abnormal
phenotype consisting mainly of plagiotropism and loss of apical dominance. Despite the variation found in somatic embryogenesis-derived
plant material, no correlation was established between genetic stability at the analyzed loci and abnormal embling phenotype,
present in 64% of the emblings. The use of microsatellites in this work was efficient for monitoring mutation events during
the somatic embryogenesis in P. pinaster. These molecular markers should be useful in the implementation of new breeding and deployment strategies for improved trees
using SE. 相似文献
12.
Trabelsi El Bahri Bouzid Sadok Bouzid Monji Elloumi Nedra Belfeleh Zina Benabdallah Abdallah Ghezel Rachida 《Journal of Plant Biology》2003,46(3):173-180
We induced somatic embryogenesis from the cotyledon segments ofOlea europaea (L) cvs. ‘Chetoui’, ‘Chemleli’, and ‘Arbequina’. Calli were established from all three cultvars on OMc media supplemented
with IBA and 2i-R The greatest success was obtained with media that contained zero or low concentrations of growth regulators.
High levels of hormones (i.e.,>0.5 mgL-1 IBA and 2i-P) inhibited embryogenesis. Embryos at different maturation stages were observed with continuously proliferating
secondary embryogenesis. Abnormally shaped embryos and teratoma were also noted. Four weeks was the optimal incubation period
for inducing embryogenesis on the auxin-containing medium. In addition, 30 to 40 gL-1 sucrose was more effective than glucose in stimulating the growth and maturation of somatic embryos. Embryogeic efficiency
was also higher when multivariate combinations of nitrogen sources (inorganic and organic nitrogen forms) were used. The plantlets
that were derived from our germinating somatic embryos were similar to those obtained from axillary buds. 相似文献
13.
Toshiro Oridate Haruhisa Atsumi Satishi Ito Hajime Araki 《Plant Cell, Tissue and Organ Culture》1992,29(1):27-30
Significant differences in somatic embryogenesis from melon seeds were observed among 18 cultivars; especially, cultivars Earl's Favorite and Barnett which produced a large number of somatic embryos. F1 seeds were obtained by reciprocal crosses between cultivars. Some lines produced a large number of somatic embryos whereas others showed no or poor embryogenic response. Most of the F1 seeds formed somatic embryos. The frequency of somatic embryogenesis decreased as compared to the parents with the highest potential. Transfer of the frequency of somatic embryogenesis from superior responding cultivars to inferior cultivars was proved. It was difficult to determine the mode of inheritance of somatic embryogenesis because there was a large variation in the range of somatic embryogenesis from F2 seeds, and cytoplasmic effect was recognized in certain combinations.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- BAP
6-benzylaminopurine 相似文献
14.
Helena Mathews C. Schopke R. Carcamo P. Chavarriaga C. Fauquet R. N. Beachy 《Plant cell reports》1993,12(6):328-333
Methods for improving the efficiency of plant recovery from somatic embryos of cassava (Manihot esculenta Crantz) were investigated by optimizing the maturation regime and incorporating a desiccation stage prior to inducing germination. Somatic embryos were induced from young leaf lobes of in vitro grown shoots of cassava on Murashige and Skoog medium with 2,4-dichlorophenoxy acetic acid. After 15 to 20 days of culture on induction medium, the somatic embryos were transferred to a hormone free medium supplemented with activated charcoal. In another 18 days mature somatic embryos became distinctly bipolar and easily separable as individual units and were cultured on half MS medium for further development. Subsequent desiccation of bipolar somatic embryos resulted in 92% germination and 83% complete plant regeneration. The plants were characterized by synchronized development of shoot and root axes. Of the non-desiccated somatic embryos, only 10% germinated and 2% regenerated plants. Starting from leaf lobes, transplantable plantlets were derived from primary somatic embryos within 70 to 80 days.Abbreviations 2,4-D
2,4 dichlorophenoxyacetic acid
- BA
Benzyl aminopurine
- GA
Giberellic acid
- MS
Murashige and Skoog
- NAA
Naphthalene acetic acid 相似文献
15.
Sánchez MC Martínez MT Valladares S Ferro E Viéitez AM 《Journal of plant physiology》2003,160(6):699-707
Experiments were performed to determine the influence of maturation medium carbohydrate content on the rates of germination and plantlet conversion (root and shoot growth) of somatic embryos from four embryogenic lines derived from leaf or internode explants of Quercus robur L. seedlings. The conversion rate was favoured by high carbohydrate content as long as the maturation medium contained at least 2% sucrose, which was necessary for healthy embryo development. Given this, sorbitol and mannitol favoured the conversion rate more efficiently than sucrose, the highest rate, 32%, being achieved by medium with 6% sorbitol and 3% sucrose. Maturation treatment did not affect the root or shoot lengths of converted embryos. In supplementary experiments, 2 weeks of gibberellic acid treatment between maturation and germination treatments did not improve germination rates, but did reduce root length and the number of leaves per regenerated plantlet. In the four embryogenic lines tested, plant recovery rate was enhanced by inclusion of benzyladenine into the germination medium following culture of the embryos on maturation medium with 6% sorbitol and 2-3% sucrose. In embryogenic systems it is important to assess the uniformity of the regenerants. Random amplified polymorphic DNA (RAPD) analysis using 32 arbitrary oligonucleotide primers was performed to study variability in DNA sequences within and between four embryogenic lines. No intraclonal nor interclonal polymorphism was detected between embryogenic lines originating from different types of explant from the same seedling, but every one of the primers detected enough polymorphism among clones originating from different plants to allow these three origins to be distinguished. No differences in DNA sequences between regenerated plantlets and their somatic embryos of origin were detected, but a nodular callus line that had lost its embryogenic capacity was found to be mutant with respect to three other clones originating from the same plantlet. This study shows that high carbohydrate levels in the maturation medium significantly increase plant conversion of oak somatic embryos, which exhibit no variation in DNA sequences when proliferated by secondary embryogenesis. 相似文献
16.
Direct somatic embryogenesis induced from cotyledons of mango immature zygotic embryos 总被引:3,自引:0,他引:3
Summary For the first time, regenerated plantlets were obtained from immature zygotic embryos of mango (Mangifera indica L.) through direct somatic embryogenesis. Pro-embryogenic mass (PEM)-like structures, which are differentiated as clusters
of globular structures, were easily induced directly from the abaxial side of cotyledons from immature fruits, 2.0–3.5 cm
diameter by a 2-wk culture period on a modified Murashige and Skoog medium with 5 mgl−1 (25μM) indole-3-butyric acid (IBA). Conversion of somatic embryos into plantlets was achieved after 4 wk of culture on the conversion
medium containing 5mgl−1 (23 μM) kinetin. Secondary somatic embryogenesis could also be obtained directly from the hypocotyls of mature primary somatic embryos
cultured on the conversion medium. In our experimental system, only minor problems were noted with browning of cultures. 相似文献
17.
Summary Calluses from five asparagus genotypes G14, G32, G171, G203, and G447 and hybrid Jersey Giant (JG) were incubated at three
temperature regimes (24, 27, and 30°C) on embryo induction medium to assess somatic embryo development and conversion to plantlets.
The calluses from three genotypes (G14, G32, and G171) were not responsive, failing to produce somatic embryos at any temperature
regime. For three responsive genotypes (G203, G447, and JG), both incubation temperature and genotype significantly affected
the numbers of somatic embryos produced. The calluses produced the most and the least numbers of total, bipolar, and globular
embryos when incubated at 27°C and 24°C, respectively. When incubated at 27°C, G203 produced the highest numbers of total
and globular embryos, 178 g−1 callus and 142 g−1 callus, respectively while G447 produced the highest number of bipolar embryos, 77 g−1 callus. Incubation temperature but not genotype significantly affected the conversion of somatic embryos to plantlets. The
somatic embryos recovered from the three responsive genotypes incubated at 27°C also converted to plantlets at the highest
frequencies, 60–63% of the bipolar embryos and 42–43% of the globular embryos converted to plantlets, while the somatic embryos
recovered from the calluses incubated at 24°C converted to plantlets at the lowest frequencies. 相似文献
18.
In planta differentiation of somatic embryos was induced in seedlings of peanut (Arachis hypogaea L.) obtained from mature seeds germinated on a medium supplemented with thidiazuron (TDZ: N-phenyl-N1- (1,2,3 thiadiazol-yl)urea). At optimum levels of TDZ (10 M), all germinating seeds produced embryogenic seedlings, and somatic embryos developed in the apical region and on the surface of cotyledons and hypocotyls. These somatic embryos matured, germinated, and formed shoots which eventually developed into whole plants. Thidiazuron-induced direct embryogenesis from morphologically intact seedlings may provide an excellent experimental system for investigating somatic embryogenesis and the morphoregulatory role of TDZ.Abbreviations 2,4-D
2,4-dichlorophenoxyacetic acid
- MS
Murashige and Skoog (1962) medium
- TDZ
thidiazuron (N-phenyl-N1(1,2,3 thiadiazol-yl)urea)
This research was supported by an operating grant from the Natural and Engineering Research Council of Canada to P.K.S. We thank Drs. J.A. Qureshi and Judith Strommer for helpful discussions, and Sangeeta Saxena for technical assistance. A gift of technical-grade thidiazuron from Nor-Am Chemical Co., Wilmington, Del., USA is gratefully acknowledged. 相似文献
19.
S. Khuri M.R. Shmoury R. Baalbaki M. Maunder S.N. Talhouk 《Biodiversity and Conservation》2000,9(9):1261-1273
Cedrus libani, the cedar of Lebanon, is a threatened conifer native to the Levant. Over 4000 years of exploitation have resulted in the fragmentation and degradation of the Lebanese cedar populations. Continued urban and agricultural development in Lebanon adds to the difficulty of effective conservation. Two protected areas have recently been established which contain two of the more important forests: a cedar dominated forest in the Shouf region and a mixed forest at Ehden. A number of other populations are protected by ministerial decrees, and there is a need for rigorous management of all the remaining populations. The application of in vitro techniques such as somatic embryogenesis may assist in the conservation of this species. We have produced somatic pro-embryos using immature zygotic tissue as explants cultured on half-strength MS medium containing an auxin and a cytokinin (10 M 2,4-D and 5 M BAP). The application of somatic embryogenesis to the Lebanese cedar would be in the propagation and preservation of selected genotypes, either those from old growth provenance for use in restoration, or those with desirable commercial or horticultural characteristics. 相似文献
20.
Summary We have developed efficient methods for plant regeneration, via both embryogenesis and organogenesis, of Smooth Cayenne pineapple,
Ananas comosus (L.) Merr. Leaf bases and core (stem) sections of in vitro shoots, produced from culture of crown tip meristem, were used as explants for plant regeneration as follows: (1) Leaf base
and core section explants cultured on Murashige and Skoog (MS) medium containing 41 μM 4-amino-3,5,6-trichloropicolinic acid (picloram, P) or thidiazuron (T)/P combinations produced embryogenic tissues. Different
types of embryogenic tissues (friable emryogenic tissue, embryogenic cell cluster, and chunky embryogenic tissue) have been
developed with varying properties in terms of growth rate and state of development. The embryogenic tissues regenerated shoots
upon culture on MS medium containing 13 μM 6-benzylaminopurine (BA) and 1μM α-naphthaleneacetic acid (NAA) followed by culture on MS medium containing 4 μM BA. (2) Crown tip meristems cultured on MS medium containing 13 μM BA followed by leaf explants cultured on MS medium with 27 μM NAA and 1 μM BA produced shoots via direct organogenesis. (3) Explants cultured on MS medium containing 5 μM T and 0.5 μM indole-3-butyric acid (IBA) produced nodular globular structures, which produced shoots upon culture on MS medium containing
1 μM BA and 1 μM gibberellic acid. Shoots obtained from all of the above methods were rooted in half-strength MS medium containing 3 μM NAA and 2.5 μM IBA. Plants were transferred to the greenhouse or shipped to Costa Rica for field trials. Somatic embryo-derived plants exhibited
21 % spininess, and organogenic-derived plants exhibited 5% spininess in the field trials. 相似文献