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1.
Junaid Aslam Abdul Mujib Zohra Fatima Maheshwar Prasad Sharma 《In vitro cellular & developmental biology. Plant》2010,46(4):348-353
Catharanthus roseus L. (G) Don. is an important dicotyledonous medicinal plant that produces anticancer compounds, which are used for the treatment
of a wide variety of cancers. We have quantified vinblastine (a major dimeric anticancer compound) in various in vitro raised tissues; embryogenic and nonembryogenic calli, three different embryogenic stages (proliferated, matured, and germinating
embryo), somatic embryo derived plantlets and in ex vitro grown plantlets by using high performance liquid chromatography. Of the various obtained callus lines and embryogenesis stages,
maximum vinblastine content was found in leaf callus and in germinating embryos. The leaves of somatic embryo-derived plantlets
contained more vinblastine than did Catharanthus leaves developed ex vitro. The yield of vinblastine was monitored for 30 wk. The production of vinblastine appeared to be age dependent and tissue
specific; the finding of our analyses is discussed in detail. 相似文献
2.
M. T. Martínez M. C. San José A. M. Vieitez M. J. Cernadas A. Ballester E. Corredoira 《Plant Cell, Tissue and Organ Culture》2017,131(2):321-333
Somatic embryogenesis from in vitro leaf and shoot apex explants excised from axillary shoot cultures established from two mature Quercus ilex trees has been developed. Somatic embryos (SE) were obtained from both explant types and genotypes evaluated, although embryogenic frequencies were influenced by the genotype, auxin concentration, and explant type. The explants were cultured on Murashige and Skoog salts and vitamins, supplemented with 500 mg L?1 casein hydrolysate (CH) and different concentrations of indole-3-acetic acid or α-naphthalene acetic acid (NAA) in combination with 2.22 µM 6-benzylaminopurine (BA). In both genotypes, shoot apex explants were more responsive than leaf explants. The best results were obtained with apex explants of clone Q3 (11%) cultured on medium with 21.48 µM NAA plus 2.22 µM BA. This combination was also effective for initiating SE from leaf explants, although the induction rates were lower (1–3%). Embryogenic lines were maintained by repetitive embryogenesis following culture of nodular embryogenic structures on Schenk and Hildebrand medium without plant growth regulators. Low embryo multiplication rates were obtained when torpedo or early cotyledonary SE were used as initial explant for embryo proliferation, or when glutamine or CH (500 mg L?1) was added to proliferation medium. For germination, cotyledonary-stage SE were isolated and stored at 4 °C for 2 months. After cold storage, SE were cultured on germination medium consisting of Gresshoff and Doy medium, supplemented with 0.44 μM BA and 20 μM silver thiosulphate. Under these conditions, plantlets were regenerated from 21 to 66.7% of the SE generated for both genotypes. 相似文献
3.
The anticancer agents vinblastine and vincristine are bisindole alkaloids derived from coupling vindoline and catharanthine, monoterpenoid indole alkaloids produced exclusively by the Madagascar periwinkle (Catharanthus roseus). Industrial production of vinblastine and vincristine currently relies on isolation from C. roseus leaves, a process that affords these compounds in 0.0003–0.01% yields. Metabolic engineering efforts to either improve alkaloid content or provide alternative sources of the bisindole alkaloids ultimately rely on the isolation and characterization of the genes involved. Several vindoline biosynthetic genes have been isolated, and the cellular and subcellular organization of the corresponding enzymes has been well studied. However, due to the leaf-specific localization of vindoline biosynthesis, and the lack of production of this precursor in cell suspension and hairy root cultures of C. roseus, further elucidation of this pathway demands the development of reverse genetics approaches to assay gene function in planta. The bipartite pTRV vector system is a Tobacco Rattle Virus-based virus-induced gene silencing (VIGS) platform that has provided efficient and effective means to assay gene function in diverse plant systems. A VIGS method was developed herein to investigate gene function in C. roseus plants using the pTRV vector system. The utility of this approach in understanding gene function in C. roseus leaves is demonstrated by silencing known vindoline biosynthetic genes previously characterized in vitro. 相似文献
4.
Qifang Pan Yu Chen Quan Wang Fang Yuan Shihai Xing Yuesheng Tian Jingya Zhao Xiaofen Sun Kexuan Tang 《Plant Growth Regulation》2010,60(2):133-141
Catharanthuse roseus is a well-known medicinal plant for its two valuable anticancer compounds: vinblastine and vincristine, which belongs to
terpenoid indole alkaloids. Great efforts have been made to study the principles of its secondary metabolic pathways to regulate
the alkaloids biosynthesis. In this article, different plant growth regulators were shortly applied to Catharanthus roseus plants during the blooming period to study their effects on the biosynthesis of vinblastine, vindoline and catharanthine.
Salicylic acid and ethylene (ethephon) treatments resulted in a significant increase of vinblastine, vindoline and catharanthine
while abscisic acid and gibberellic acid had a strongly negative influence on the accumulation of the three important alkaloids.
Methyl jasmonate showed no great effect on the production of these valuable alkaloids. Chlormequat chloride highly enhanced
the accumulation of vinblastine but greatly decreased the contents of vindoline and catharanthine. 相似文献
5.
Factors affecting microspore embryogenesis of cow cockle (Saponaria vaccaria) were evaluated including donor plant growing conditions, genotype, bud size, density, medium composition, and culture conditions.
Of the two donor plant (day/night) temperature regimes evaluated (10/5°C and 20/15°C), plants grown at 20/15°C were the most
embryogenic. An embryogenic frequency of greater than 350 embryos/100 buds was observed in the most embryogenic genotype,
cv. ‘White Beauty’. Buds from 3–9 mm in length were evaluated for their embryogenic potential; buds that were 4–7.9 mm produced
the most embryos/100 buds. Of all the media compositions evaluated, NLN medium with 15% sucrose resulted in the most embryos.
Cow cockle microspores required an initial period of 32°C for 3 days for production of microspore-derived embryos (MDEs). 相似文献
6.
Raffaella Filippini Rosamaria Caniato Francesca Dalla Vecchia Elsa M. Cappelletti Lucia Puricelli Anna Piovan 《Plant biosystems》2013,147(2):179-184
ABSTRACT A new protocol to obtain an embryogenic cell line from cultured seedling explants of Catharanthus roseus is described. In order to assess the relationship between tissue differentiation and secondary metabolite biosynthesis, the biosynthetic capabilities (alkaloid production) of an embryogenic cell line and two non-embryogenic C. roseus strains were comparatively examined. Faster cell growth rate was associated with higher alkaloid production in the embryogenic cell line. The kinetics of ajmalicine and serpentine production by the three cell lines is also reported. 相似文献
7.
Bin Huang Sharon Bird Roger Kemble Brian Miki Wilf Keller 《In vitro cellular & developmental biology. Plant》1991,27(1):28-31
Summary Microscope cultures ofBrassica napus cv. Topas undergo high frequency embryogenesis in vitro; however, the majority of microspore-derived embryos do not develop
directly into plants but usually undergo abnormal development including the formation of secondary embryos on the hypocotyls.
The present studies show that older embryos or embryos cultured at higher temperature (25° C) were more likely to undergo
secondary embryogenesis whereas embryos cultured at 20° C or pretreated at 5° to 10° C for 28 days developed more readily
into normal plants. Compared with embryos cultured at 25° C, those cultured at 20° C gave a threefold increase in normal plant
production. Pretreatments at cooler temperatures (5° to 10° C) resulted in an additional two-to threefold increase in the
recovery of normal plants. Higher osmoticum during pretreatment improved embryo survival at low temperatures but generally
inhibited normal plant development. Abscisic acid was ineffective or deleterious. 相似文献
8.
In this study, loop-mediated isothermal amplification (LAMP)-based molecular marker was developed for authentication of Catharanthus roseus, a medicinal plant. Samples of this plant were collected from different geographical locations in India. Random amplified
polymorphic deoxyribonucleic acid (DNA) analysis of collected samples was carried out with 25 random primers. A 610-bp DNA
fragment, common to all accessions, was eluted, cloned, and sequenced. Four LAMP primers were designed on the basis of sequence
of 610 bp DNA fragment. LAMP reaction, containing 10× Bst DNA polymerase reaction buffer, Bst DNA polymerase, four in-house designed primers, dNTPs, MgSO4, and betaine, was incubated at 65°C for 1 h. The resulting amplicon was visualized by adding SYBR Green I to the reaction
tube. The data showed confirmatory results. Since the assay method is simple, sensitive, and cost-effective, it is a feasible
method for identifying and authentication of C. roseus. 相似文献
9.
Ana L. D. M. Furlanetto Silvia M. S. C. Cadena Glaucia R. Martinez Beatriz Ferrando Tinna Stevnsner Ian M. Mller 《Physiologia plantarum》2019,166(2):513-524
We evaluated the effect of global warming on Araucaria angustifolia (Bert.) O. Kuntze, a critically endangered native tree of Southern Brazil, by studying the effects of short‐term high temperature treatment on cell viability, respiration and DNA repair of embryogenic cells. Compared with control cells grown at 25°C, cell viability was reduced by 40% after incubation at 30 and 37°C for 24 and 6 h, respectively, while 2 h at 40 and 42°C killed 95% of the cells. Cell respiration was unaffected at 30–37°C, but dramatically reduced after 2 h at 42°C. The in vitro activity of enzymes of the base excision repair (BER) pathway was determined. Apurinic/apyrimidine endonuclease, measured in extracts from cells incubated for 2 h at 42°C, was completely inactivated while lower temperatures had no effect. The activities of three enzymes of the mitochondrial BER pathway were measured after 30‐min preincubation of isolated mitochondria at 25–40°C and one of them, uracil glycosylase, was completely inhibited at 40°C. We conclude that cell viability, respiration and DNA repair have different temperature sensitivities between 25 and 37°C, and that they are all very sensitive to 40 or 42°C. Thus, A. angustifolia will likely be vulnerable to the short‐term high temperature events associated with global warming. 相似文献
10.
11.
Atar Selouane Driss Bouya Ahmed Lebrihi C. Decock Amina Bouseta 《Journal of microbiology (Seoul, Korea)》2009,47(4):411-419
The effects of temperature, water activity (aw), incubation time, and their combinations on radial growth and ochratoxin A (OTA) production of/by eight Aspergillus niger aggregate strains (six A. tubingensis and two A. niger) and four A. carbonarius isolated from Moroccan grapes were studied. Optimal conditions for the growth of most studied strains were shown to be at
25°C and 0.95 aw. No growth was observed at 10°C regardless of the water activity and isolates. The optimal temperature for OTA production
was in the range of 25°C∼30°C for A. carbonarius and 30°C∼37°C for A. niger aggregate. The optimal aw for toxin production was 0.95∼0.99 for A. carbonarius and 0.90∼0.95 for A. niger aggregate. Mean OTA concentration produced by all the isolates of A. niger aggregate tested at all sampling times shows that maximum amount of OTA (0.24 μg/g) was produced at 37°C and 0.90 aw. However, for A. carbonarius, mean maximum amounts of OTA (0.22 μg/g) were observed at 25°C and 0.99 aw. Analysis of variance showed that the effects of all single factors (aw, isolate, temperature and incubation time) and their interactions on growth and OTA production were highly significant. 相似文献
12.
Effects of non-specific ethylene biosynthesis inhibitors: salicylic acid (SA) and aminoethoxyvinylglycine (AVG), and of specific
inhibitors of ethylene binding to receptors: 1-methylcyclopropene (1-MCP) and 2,5-norbornadiene (NBD) applied during proliferation
and differentiation phases of indirect somatic embryogenesis (SE) of Medicago sativa L. cv. Rangelander on embryogenic suspension growth, embryo production, development, and ability to germinate and convert
were studied. Application of SA and AVG alone or together at concentrations from 1 to 500 μM in B5g liquid medium during the proliferation phase had an inhibitory effect on ethylene production and embryogenic suspension
growth. Additionally, it caused a drastic reduction in production of embryos and their development on BOi2Y solid differentiation
medium. The inhibitory effect of SA was more visible than that of AVG. In addition, disturbance of ethylene biosynthesis during
the proliferation phase of SE resulted in diminished lateral germination and conversion of cotyledonary embryos on MS solid
medium. Moreover, blocking of ethylene receptors by 1-MCP during the proliferation phase also inhibited ethylene production
and embryogenic suspension growth and reduced embryo production during differentiation. MCP almost completely inhibited development
of cotyledonary embryos. At the same time, development of more embryos was arrested at the globular stage, and the number
of abnormal embryos almost doubled. Similarly, addition of 1-MCP or NBD to the ambient atmosphere during the differentiation
phase evidently arrested the development of embryos and, consequently, their ability to germinate and convert on MS regeneration
medium. All the results presented above demonstrated that not only ethylene biosynthesis, but also ethylene action is involved
in the control of individual phases of SE in Medicago sativa L. cv. Rangelander. And what is more, disturbance of these processes during distinct phases of SE adversely affects vigor
of the somatic embryos obtained. 相似文献
13.
An efficient somatic embryogenesis system has been established in Catharanthus roseus (L.) G. Don in which primary and secondary embryogenic calluses were developed from hypocotyls and primary cotyledonary somatic
embryos (PCSEs), respectively. Two types of calluses were different in morphology and growth behaviour. Hypocotyl-derived
embryogenic callus (HEC) was friable and fast-growing, while secondary callus derived from PCSE was compact and slow-growing.
HEC differentiated into somatic embryos which proliferated quickly on medium supplemented with NAA (1.0 mg l−1) and BA (1.5 mg l−1). Although differentiation and proliferation of somatic embryos were faster in primary HEC, maturation and germination efficiency
were better in somatic embryos developed from primary cotyledonary somatic embryo-derived secondary embryogenic callus (PCSEC).
At the biochemical level, two somatic embryogenesis systems were different. Both primary and secondary/adventive somatic embryogenesis
and the role of plant growth regulators in two modes of somatic embryo formation have been discussed. 相似文献
14.
Sottomayor M. Lopes Cardoso I. Pereira L.G. Ros Barceló A. 《Phytochemistry Reviews》2004,3(1-2):159-171
The leaves of Catharanthus roseus (L.) G. Don produce the first natural drugs used in cancer therapy – the dimeric terpenoid indole alkaloids vinblastine and vincristine. The study of C. roseus further revealed two other terpenoid indole alkaloids with important pharmacological activity: ajmalicine, used as an antihypertensive, and serpentine, used as sedative. The biosynthetic pathway of the medicinal alkaloids has been investigated in much detail and a number of steps are now well characterized at the enzyme and gene level and, recently, several regulatory genes have also been isolated and characterized. Since early studies of the biosynthesis of vinblastine, during the 1970s and 1980s, the dimerization reaction has attracted much attention due to its possible regulatory importance and potential application for the semi synthetic production of the dimeric alkaloids. After initial, inconclusive work suggesting the involvement of peroxidase-like enzymes, the search for the dimerization enzyme in leaf tissue detected a single dimerization activity credited to the single class III plant peroxidase present in the leaves of the plant – the basic isoenzyme CRPRX1. The enzyme was purified to homogeneity, the respective cDNA and genomic sequences were characterized, and a channeling mechanism was proposed for the peroxidase-mediated-vacuolar synthesis of the first dimeric alkaloid intermediate, α-3′,4′-anhydrovinblastine. On the other hand, the oxidation of ajmalicine into serpentine has been attributed to basic peroxidase isoenzymes localized in the vacuole of C. roseus cells. An overview of the work implying class III plant peroxidases in the biosynthesis of terpenoid indole alkaloids in C. roseus is presented here. Abbreviations: CRPRX1 –Catharanthus roseus peroxidase 1; DAB – diaminobenzidine; IEF – isoelectric focusing; UV – ultraviolet. 相似文献
15.
Optimized somatic embryogenesis in Pinus strobus L. 总被引:5,自引:0,他引:5
Krystyna Klimaszewska Yill-Sung Park Cathy Overton Ian Maceacheron Jan M. Bonga 《In vitro cellular & developmental biology. Plant》2001,37(3):392-399
Summary Somatic embryogenesis (SE) initiation in Pinus strobus was optimized by the manipulation of plant growth regulator (PGR) concentrations in the culture medium. Modified Litvay medium
(MLV) of Litvay et al. (1985) supplemented with lower than routinely used PGR concentration increased initiation of established
embryogenic cultures from approximately 20 to 53%. The original developmental stage of zygotic embryos had a pronounced effect
on the SE response. The optimum stage was the pre- to shortly post-cleavage stage. A substantial genetic influence on initiation
of SE was indicated by a significant variance component due to families. Genotype X collection date and genotype X media interactions
had large effects on initiation of SE. The PGR levels in the culture medium prior to maturation had a significant effect on
subsequent production of mature somatic embryos. Embryogenic tissue initiated and proliferated on medium with a low level
of PGR consistently produced a high number of somatic embryos, indicating that optimized initiation protocol also enhanced
somatic embryo production. Somatic embryos of 93 embryogenic lines (representing five families) that were initiated on media
with different PGR concentrations were converted to plants at an overall frequency of 76%, and grown in the greenhouse. With
these improved protocols, application of P. strobus SE in commercial clonal forestry is feasible as an alternative to traditional breeding and reforestation. 相似文献
16.
Waldemar Buchwald Irena Dedio Jan Kozłowski Barbara Łata 《Phytochemistry Reviews》2007,6(2-3):413-417
Under climatic conditions of Poland, Catharanthus roseus may be grown in the open field only as annual crop, using transplants raised in a greenhouse or in a plastic tunnel. An effective
method of hydroponic culture has been elaborated and tested. Herbal material obtained in hydroponics showed considerable concentration
of alkaloids in different plant parts. Seeds of C. roseus may also be produced under these conditions. Characteristics of seeds as well as the effect of different methods of their
storage are presented. Seeds stored in paper bags at room temperature may retain germinability for 7 years and those stored
in a refrigerator, at 5°C, up to 15 years. Storage in a freezer, at -10°C, resulted in a quick and sharp reduction of germination
rate. 相似文献
17.
Fecundity and feeding of two introduced sibling biological control species, Galerucella calmariensis and G. pusilla (Coleoptera: Chrysomelidae) on purple loosestrife, Lythrum salicaria L. (Lythraceae) were compared at constant temperatures of 12.5, 15, 20, 25, and 27.5 °C. Larval feeding was also carried
out at 30 °C, but at this temperature, larvae developed only to the L2 stage and none pupated. Thus, data for this temperature
were not used in the analysis. There were significant species × temperature interactions in fecundity. Of the two species,
Galerucella pusilla laid more eggs. Although egg production of both species was lowest at 12.5 °C and increased to 20 °C, at higher temperatures,
the two species reacted differently. From 25 to 27.5 °C, egg production decreased for G. pusilla, but G. calmariensis fecundity peaked at 27.5 °C. Significant temperature × species × life-stage interactions were also observed in feeding.
For each species, the amount of feeding varied with temperature and stage of development. Galerucella pusilla adults consumed more foliage at 15, 20, and 27.5 °C. However, at 12.5 °C G. calmariensis adults fed more than G. pusilla. G. pusilla larvae consumed an average of 25% less foliage than G. calmariensis. The lower larval consumption of G. pusilla suggests that when food is limited, G. pusilla larvae may have a higher survival rate because of its ability to complete larval development with less food and produce more
progeny due to its greater fecundity. When food is not limited neither species would have a competitive advantage and both
species could coexist temporally and spatially. However, since G. calmariensis larvae consumed more leaf material, the larval stage of this species would have a greater impact on purple loosestrife than
G. pusilla. 相似文献
18.
Effect of plant growth regulators, explant size, season of explant collection, temperature (20, 25 and 30 °C) and photoperiod
on in vitro lotus (Nelumbo nucifera Gaertn.) shoot formation and growth were examined. Shoots formation was greatly influenced by growth regulators, explant
size and season of explant collection. The maximum number of shoots were induced from bud explants on Murashige and Skoog
(MS) medium containing 4.44 μM benzyladenine (BA) + 0.54 μM α-naphthalene acetic acid (NAA). Explants formed by bud of one
expanded and one unexpanded leaf, which was collected in spring gave encouraging results of shoot production. Higher temperature
favoured shoot induction and subsequent growth was much better at 25 °C compared to that at 20 and 30 °C. 相似文献
19.
Effects of abscisic acid (ABA) and methyl jasmonate (MeJA) on ethylene production, ACC oxidase (ACO) activity, and content
of 1-aminocyclopropane-1-carboxylic acid (ACC) during indirect somatic embryogenesis (SE) of Medicago sativa L. were studied. ABA and MeJA, at 50 μM, were applied during the induction, proliferation, or differentiation phase. ABA
decreased ethylene production at the beginning of callus and SE induction and during the differentiation of somatic embryos.
The hormone inhibited ACO activity in explants with overgrowing callus during the first two weeks of induction, in embryogenic
suspension and also in differentiating embryos. The ACC content was reduced by ABA in callus at the end of SE induction, in
embryogenic suspension and in globular embryos, but elevated in cotyledonary embryos. MeJA had no significant effect on ethylene
production during M. sativa SE, despite the fact, that it inhibited ACO activity during the first two weeks of induction and in torpedo and cotyledonary
embryos. The ACC content was increased by MeJA in 14-day-old callus and embryogenic suspension but was inhibited in globular
embryos. Both ABA and MeJA seem to be involved in the regulation of ethylene biosynthesis during distinct phases of SE in
M. sativa. It might be considered that exogenous ABA, more probably than MeJA, exerts its inhibitory effect on M. sativa somatic embryo formation by modifying ethylene production. 相似文献
20.
Zhiyong Pan Shiping Zhu Rui Guan Xiuxin Deng 《Plant Cell, Tissue and Organ Culture》2010,103(2):145-153
The compound 2,4-Dicholorophenoxyacetic acid (2,4-D) is an important growth regulator which is used in the majority of embryogenic
cell and tissue culture systems. However, 2,4-D also appears to have a negative effect on growth and development of plant
tissues and organs cultured in vitro. For example, 2,4-D exerts inhibition on in vitro somatic embryo initiation and/or development
of most citrus species. To understand the molecular mechanism by which 2,4-D inhibits somatic embryogenesis (SE), proteomic
changes of Valencia sweet orange (Citrus sinensis) embryogenic callus induced by treatments with a high concentration of 2,4-D (6 mg l−1) was investigated. Nine 2,4-D-responsive proteins were identified, of which eight were up-regulated and one was down-regulated.
Interestingly, three of the eight up-regulated proteins were osmotic stress-associated, suggesting that 2,4-D induced osmotic
stress in Valencia embryogenic callus. This speculation was supported by results from our physiological studies: 2,4-D treated
callus cells exhibited increased cytoplasm concentration with a significant reduction in relative water content (RWC) and
an obvious increase in levels of two osmolytes (proline and soluble sugar). Taken together, our results suggested that 2,4-D
could inhibit somatic embryo initiation by, at least in part, inducing osmotic stress to citrus callus cells. 相似文献