scholarly journals Enhanced Resistance to Verticillium dahliae in Transgenic Strawberry Plants Expressing a Lycopersicon chilense Chitinase Gene

2003 ◽  
Vol 128 (5) ◽  
pp. 747-753 ◽  
Author(s):  
Vida Chalavi ◽  
Zohreh Tabaeizadeh ◽  
Pierre Thibodeau
Keyword(s):  
Verticillium Dahliae ◽  
Constitutive Expression ◽  
Chitinase Gene ◽  
High Rate ◽  
Northern Analysis ◽  
35S Promoter ◽  
Indirect Organogenesis ◽  
High Tendency ◽  
Lycopersicon Chilense ◽  
Growth Chamber Studies

A chitinase gene (pcht28), isolated from Lycopersicon chilense, was transferred into `Joliette' strawberry using a stipule regeneration method and Agrobacterium-mediated gene transfer technique. Stipules showed a high rate of shoot regeneration (>90%) through direct or indirect organogenesis. Stipules were cocultured with a transformation plasmid in which pcht28 was under the control of the CAMV 35S promoter. A high tendency in production of chimaeric shoots was observed. Shoots which did not show any sign of bleaching after several subcultures in kanamycin containing medium were considered as stable transformants. These shoots were successfully rooted in the presence of 50 μg·mL-1 kanamycin. Transgenic nature of the plants was confirmed by PCR as well as Southern blot analysis. Constitutive expression of the chitinase gene was demonstrated by Northern analysis. In growth chamber studies, the transgenic strawberry plants which expressed pcht28 had significantly higher resistance to Verticillium dahliae as compared to nontransgenic controls. These results demonstrate that pcht28 plays a role in defense against this fungal disease in strawberry.

scholarly journals Transfer of Lycopersicon chilense Chitinase Gene to Cultivated Tomato (L. esculentum)

HortScience ◽  
1995 ◽  
Vol 30 (4) ◽  
pp. 878G-878
Author(s):  
Zahra Agharbaoui ◽  
Long Xi Yu ◽  
Viano Poysa ◽  
Zohreh Tabaeizadeh
Keyword(s):  
Chitinase Gene ◽  
Indirect Organogenesis ◽  
Tomato Cultivar ◽  
Wild Tomato Species ◽  
Lycopersicon Chilense ◽  
Tomato Species ◽  
Npt Ii ◽  
Selection Medium ◽  
Polymerase Chain ◽  
Npt Ii Gene

We have isolated a drought-induced chitinase gene from L. chilense, a wild tomato species. Owing to our interests in genetic improvement of cultivated tomato, we have transferred the L. chilense chitinase gene to this species. The transformation plasmid constructed contained the coding sequence of L. chilense chitinase gene linked to CaMVS35 promoter as well as the NPT II gene linked to nopaline synthetase promoter. The leaf disk transformation regeneration technique was applied to one commercial tomato cultivar and four inbred lines. Shoots were produced on the selection medium through direct or indirect organogenesis. Plantlets that have been rooted on kanamycin-containing medium were transferred to soil where they grew to maturity and produced flowers and fruit. The transgenic nature of some of the analyzed plants was confirmed by polymerase chain reaction. Research is continuing to evaluate transgenic plants with regard to their level of tolerance to phytogenic fungi.

Culture Concordance in Different Sections of the Metatarsal Head: Interpretations of Microbiological Results

2021 ◽  
pp. 153473462110037
Author(s):  
Raul Juan Molines-Barroso ◽  
Esther García-Morales ◽  
David Sevillano-Fernández ◽  
Yolanda García-Álvarez ◽  
Francisco J. Álvaro-Afonso ◽  
...  
Keyword(s):  
Diabetic Foot ◽  
Bone Biopsy ◽  
Bacterial Load ◽  
False Negative ◽  
Metatarsal Bone ◽  
High Rate ◽  
Microbiological Analysis ◽  
High Tendency ◽  
Metatarsal Bones ◽  
Bone Biopsies

Microbiological cultures of per-wound bone biopsies have shown a lack of correlation and a high rate of false-negative results when compared with bone biopsy cultures in diabetic foot osteomyelitis. The selection of samples from the area of active osteomyelitis, which contains a complete census of the microorganisms responsible for the infection, is essential to properly guide antimicrobial treatment. We aimed to comparatively evaluate the quantitative and qualitative cultures taken from different areas, in metatarsal heads resected for osteomyelitis. For this purpose, we consecutively selected 13 metatarsal heads from 12 outpatients with plantar ulcers admitted to our diabetic foot unit. Metatarsal heads were divided transversally into 3 portions: plantar (A), central (B), and dorsal (C), and the 39 resulting samples were cultured. Qualitative and quantitative microbiological analysis was performed, and the isolated species and bacterial load, total and species specific, were compared between the 3 metatarsal bone segments. The primary outcome of the study was the bacterial diversity detected in the different bone sections. Cultures were positive in 12 of the 13 included metatarsal heads (92%). A total of 34 organisms were isolated from all specimens. Ten of the 12 cultures (83%) were polymicrobial. Ten of the 13 metatarsal heads (77%) had identical microbiological results in each of the 3 bone sections. The largest number of microorganisms was found in the central section. The overall concordance between sections was 91%. The predominant microorganisms were coagulase-negative staphylococci (41%). Statistical differences were not found in the bioburden between sections (range 3.25-3.41 log10 colony-forming unit/g for all sections; P = .511). The results of our study suggest that microorganisms exhibit a high tendency to spread along the metatarsal bone and that the degree of progression along the bone is species dependent. The central portions of metatarsal bones tend to accumulate a higher diversity of species. Thus, we recommend this area of bone for targeted biopsy in patients with suspected osteomyelitis.

scholarly journals Transfer of human proinsulin gene into Cucumber (Cucumis sativus L.) via agrobacterium method

Genetika ◽  
2017 ◽  
Vol 49 (2) ◽  
pp. 717-728
Author(s):  
Kameh Abookazemi ◽  
Javaran Jalali ◽  
Mehdi Mohebodini ◽  
Akbar Vaseghi
Keyword(s):  
Transgenic Plants ◽  
Molecular Farming ◽  
Adult Population ◽  
Advanced Technology ◽  
High Rate ◽  
35S Promoter ◽  
Dot Blot ◽  
Hormone Production ◽  
Human Proinsulin ◽  
Number Of Patients

Nowadays, approximately 5.8% in adult population around the world are suffering by diabetes. It can be caused by an increase in risk factors such as being overweight. Also it has been estimated that the number of patients will be doubled in near future and the demands for insulin hormone will be growing up by 3 to 4 % annually. Therefore, it?s necessary to develop new methods for hormone production with high rate of capacity in future. By advanced technology of transgenic DNA, the transgenic plants are introduced as an attractive system for expression and production of many kinds of pharmaceutical proteins. In this study, we investigated transfer of Human Proinsulin Gene into the Cucumber (Cucumissativus L.). Transgenic cucumber could be a great prospect for future source of eatable insulin pharmaceutical drugs to be taken by patients.Agrobacterium tumefaciensstrain LBA4404 carrying proinsulin genes with CaMV 35S promoter was used for the transformation purpose. The transgenic plants were analyzed by PCR, RT-PCR, SDS-PAGE, Dot blot and Electrochemiluminescence techniques. Production of proinsulin in cucumber could be a great prospect in molecular farming of human proinsulin.

Regulation of macrophage colony-stimulating factor in liver fat-storing cells by peptide growth factors

1992 ◽  
Vol 262 (4) ◽  
pp. C876-C881 ◽  
Author(s):  
M. Pinzani ◽  
H. E. Abboud ◽  
L. Gesualdo ◽  
S. L. Abboud
Keyword(s):  
Growth Factor ◽  
Constitutive Expression ◽  
Liver Fat ◽  
Northern Analysis ◽  
Mrna Levels ◽  
Colony Stimulating Factor ◽  
Term Survival ◽  
Macrophage Colony Stimulating Factor ◽  
Macrophage Colony ◽  
Stimulating Factor

Macrophage colony-stimulating factor (M-CSF) selectively promotes mononuclear phagocyte survival, proliferation, and differentiation. The production of this factor within the liver may be necessary to support the relatively long-term survival of circulating monocytes as they migrate into tissues and differentiate into macrophages. We studied the constitutive expression and the effects of platelet-derived growth factor (PDGF), basic fibroblast growth factor (bFGF), and epidermal growth factor (EGF) on M-CSF mRNA levels and secretion of M-CSF in murine liver fat-storing cells (FSC), vascular pericytes likely involved in the development of liver fibrosis. By Northern analysis, using a murine M-CSF cDNA, FSC constitutively express two major transcripts of 4.4 and 2.2 kb, similar to those detected in mouse L cells, used as a control. Exposure to 10 ng/ml PDGF or bFGF increased M-CSF mRNA levels. Peak effects were observed at 3 and 6 h for PDGF and bFGF, respectively, returning to baseline levels by 12 h. Under basal conditions, detectable amounts of M-CSF, measured by radioimmunoassay, were found in cell supernatants conditioned for 8 and 24 h. PDGF and bFGF markedly stimulated the release of M-CSF as early as 8 h, an effect persisting for at least 24 h. These findings suggest that liver FSC release M-CSF upon stimulation by PDGF and bFGF and may contribute to the activation of resident or infiltrating cells in inflammatory liver diseases.

scholarly journals Constitutive expression of Mpl ligand transcripts during thrombocytopenia or thrombocytosis

Blood ◽  
1996 ◽  
Vol 88 (7) ◽  
pp. 2578-2584 ◽  
Author(s):  
K Cohen-Solal ◽  
JL Villeval ◽  
M Titeux ◽  
S Lok ◽  
W Vainchenker ◽  
...  
Keyword(s):  
Splice Variants ◽  
Mrna Level ◽  
Constitutive Expression ◽  
Experimental Models ◽  
Northern Analysis ◽  
Transcriptional Level ◽  
Severe Thrombocytopenia ◽  
Wild Type ◽  
Mrna Isoforms ◽  
Liver And Kidney

Mpl ligand (thrombopoietin [TPO]) is the physiological regulator of platelet production. In mice, mRNA encoding the Mpl ligand (Mpl-L) is predominantly found by Northern blot analysis in the liver and kidney. To investigate the mode of regulation of the Mpl-L gene, we have developed several experimental models of severe thrombocytopenia differing in their kinetics and an opposite model of chronic thrombocytosis. Northern analysis performed at various times after induction of a thrombocytopenic state demonstrates that, whatever the number of circulating platelets, no change in Mpl-L mRNA level occurs in liver and kidney. By ribonuclease protection assays, we analyzed the ratios between mRNAs coding for the wild-type Mpl-L form and various splice variants encoding inactive or nonsecreted Mpl-L proteins. No modification in levels of these various isoforms was detected confirming the data of a previous report. Because the highest level of Mpl-L bioactivity in sera was observed only in mice with drastically reduced numbers of both platelets and megakaryocytes, these results further suggest that not only platelets, but also megakaryocytes, must be involved in the regulation of the level of circulating Mpl-L. In addition, we show that no downregulation of wild-type Mpl-L mRNA and no change in the ratio of Mpl-L mRNA isoforms were detected in mice in which a chronic thrombocytosis was induced. Together, these different models extend and further confirm that the regulation of Mpl-L does not occur at a transcriptional level or by a modulation in the ratios of Mpl-L mRNA isoforms.

Optimization of particle bombardment parameters for DNA delivery into the male flowers of banana

Biologia ◽  
2014 ◽  
Vol 69 (7) ◽  
Author(s):  
Fatemeh Mahdavi ◽  
Maziah Mahmood ◽  
Normah Noor
Keyword(s):  
Particle Bombardment ◽  
Fluorescent Protein ◽  
Constitutive Expression ◽  
Vacuum Pressure ◽  
35S Promoter ◽  
Helium Pressure ◽  
Gfp Expression ◽  
Gfp Gene ◽  
Male Flowers ◽  
Green Fluorescent

AbstractThe possibility of increasing the efficiency of banana transformation was investigated by particle bombardment of the male flowers of banana plants for constitutive expression of gfp gene. The effects of particle bombardment parameters, such as acceleration pressure, bombardment distance, chamber vacuum pressure, gold microcarrier size, gold quantity, DNA quantity, number of bombardments and pre-culture were examined. Single cauliflower-like bodies (CLBs) clusters, induced from meristemic parts of Musa sapientum cv. Nangka (AAB) male flowers, were bombarded by pCambia1304 plasmid carrying gfp gene driven by the CaMV 35S promoter. Optimal transient expression of green-fluorescent protein (GFP) was obtained when the three-day old cultured tissues were bombarded two times at 1100 psi helium pressure. However, the highest GFP expression was observed when 9 cm was applied as bombardment distance with 28 mmHg chamber vacuum pressure. Gold particle with 1 μm diameter at 60 μg/μL concentrations coated with 1.5 μg/μL of DNA have been used as the optimum bombardment parameter since GFP expression was significantly different compared to other conditions. Application of optimized condition proved effective for the generation of stable transgenic banana plants. PCR and southern blot analyses confirmed the presence and integration of gfp gene in genomic DNA of transformed plants. Transformation frequency achieved with the optimized protocol was 7.5% which was significantly higher than the conventional protocol.

scholarly journals Historical Contingency Causes Divergence in Adaptive Expression of the lac Operon

2021 ◽  
Author(s):  
Kedar Karkare ◽  
Huei-Yi Lai ◽  
Ricardo B R Azevedo ◽  
Tim F Cooper
Keyword(s):  
Promoter Region ◽  
Potential Benefit ◽  
Constitutive Expression ◽  
High Rate ◽  
Mutation Rates ◽  
Lac Operon ◽  
Historical Contingency ◽  
Fitness Effect ◽  
Fluctuating Environments ◽  
Laci Repressor

Abstract Populations of Escherichia coli selected in constant and fluctuating environments containing lactose often adapt by substituting mutations in the lacI repressor that cause constitutive expression of the lac operon. These mutations occur at a high rate and provide a significant benefit. Despite this, eight of 24 populations evolved for 8,000 generations in environments containing lactose contained no detectable repressor mutations. We report here on the basis of this observation. We find that, given relevant mutation rates, repressor mutations are expected to have fixed in all evolved populations if they had maintained the same fitness effect they confer when introduced to the ancestor. In fact, reconstruction experiments demonstrate that repressor mutations have become neutral or deleterious in those populations in which they were not detectable. Populations not fixing repressor mutations nevertheless reached the same fitness as those that did fix them, indicating that they followed an alternative evolutionary path that made redundant the potential benefit of the repressor mutation, but involved unique mutations of equivalent benefit. We identify a mutation occurring in the promoter region of the uspB gene as a candidate for influencing the selective choice between these paths. Our results detail an example of historical contingency leading to divergent evolutionary outcomes.

scholarly journals AAC as a Potential Target Gene to Control Verticillium dahliae

2016 ◽  
Author(s):  
Xiaofeng Su ◽  
Latifur Rehman ◽  
Huiming Guo ◽  
Xiaokang Li ◽  
Hongmei Cheng
Keyword(s):  
Verticillium Dahliae ◽  
Uv Light ◽  
Fungal Growth ◽  
35S Promoter ◽  
Fungal Development ◽  
Knockout Mutants ◽  
High Concentrations ◽  
Novel Strategy ◽  
Fungal Genes ◽  
Type V

Verticillium dahliae invades the roots of host plants and causes vascular wilt, which seriously diminishes the yield of cotton and other important crops. The protein AAC (ADP, ATP carrier) is responsible for transferring ATP from the mitochondria into the cytoplasm. When V. dahliae protoplasts were transformed with short interfering RNAs (siRNAs) targeting the VdAAC gene, fungal growth and sporulation were significantly inhibited. To further confirm a role for VdAAC in fungal development, we generated knockout mutants (ΔVdACC), which were hypersensitive to stresses such as UV light and high concentrations of NaCl or sorbitol. Compared with wild-type V. dahliae  (Vd wt), ΔVdAAC was impaired in germination and virulence; these impairments were rescued in the complementary strains (ΔVdAAC-C). Moreover, when an RNAi construct of VdAAC under the control of the 35S promoter was used to transform Nicotiana benthamiana, the expression of VdAAC was downregulated in the transgenic seedlings, and they had elevated resistance against V. dahliae. The results of this study suggest that VdAAC contributes to fungal development, virulence and response to stresses and is a promising candidate gene to control V. dahliae. In addition, RNAi is a highly efficient way to silence fungal genes and provides a novel strategy to improve disease resistance in plants.

Rapid in vitro Propagation of Dodonaea viscosa (Sapindaceae) Through Axillary Bud Multiplication and Indirect Organogenesis

2015 ◽  
Vol 22 (1) ◽  
pp. 33-36
Author(s):  
A. Benniamin ◽  
G. Jothi ◽  
M. Sundari
Keyword(s):  
Acetic Acid ◽  
In Vitro Propagation ◽  
High Rate ◽  
Axillary Bud ◽  
Naphthalene Acetic Acid ◽  
Ms Medium ◽  
Shoot Induction ◽  
Dodonaea Viscosa ◽  
Indirect Organogenesis

Protocols of axillary bud multiplication were established for Dodonaea viscosa (Sapindaceae). Murashig & Skoog’s medium with 0.2 mg/l BAP with 0.01 Naphthalene acetic acid induced high rate of shoot induction and an average of five shoots per node. Subsequent culture enhanced the number of shoots. Callus initiated from the basal cut end explants differentiated in to more than 10 shoots on MS Medium with 0.4mg/l Benzylaminopurine and 0.02mg/l Indole Butric Acid. Eighty per cent of the rooted shoots survived when transferred to greenhouse and subsequently to the field.

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