This document reports on the partial characterization of Prunus Necrotic Ringspot Virus (PNRSV) isolated from apple trees in Egypt. Apple trees infected with PNRSV showed chlorotic and necrotic leaf spots and shoot holes. Serological tests and RT-PCR confirmed the presence of PNRSV. Electron microscopy of the purified viral preparation showed spherical virions approximately 26nm in diameter, consistent with members of the genus Ilarvirus. This study identifies PNRSV as the causal agent of apple disease symptoms in Egypt.
RNAi as a novel technology in pest control: current status and challenges - O...OECD Environment
10-12 April 2019: The OECD Conference on RNAi based pesticides provided an overview on the current status and future possibilities for the regulation of externally applied dsRNA-based products that are proposed for use as pesticides. The event facilitated exchanges between policy makers, academia, industry on their implications in health, environment, and regulation.
Environmental dissipation of dsRNA in soil, aquatic systems and plants - Pame...OECD Environment
10-12 April 2019: The OECD Conference on RNAi based pesticides provided an overview on the current status and future possibilities for the regulation of externally applied dsRNA-based products that are proposed for use as pesticides. The event facilitated exchanges between policy makers, academia, industry on their implications in health, environment, and regulation.
To study of the genetic variations among the Azospirillum lipoferu isolates u...ijsrd.com
Among free-living microorganisms, which can be practically used in agriculture, bacteria from the Azospirillum genus as well as other endophytes are nowadays thought of as the most active component of associative dinitrogen fixation. The investigation was carried out to study the characterization of Azospirillum lipoferu found in the soils of the ten agro-climatic zones which Karnataka, is classified. By using RAPD markers, 75 bands were scored out of which 78.6 % were found to be polymorphic. Statistical analysis of RAPD data enabled the classification of 10 Azospirillum isolates into two major groups. . In this, the cluster analysis based on 75 RAPD bands revealed that the ten A. lipoferu isolates examined clustered at a linkage distance of about 40 units on the dendrogram. There was no correlation between RAPD and geographical origin of isolates.
RNAi as a novel technology in pest control: current status and challenges - O...OECD Environment
10-12 April 2019: The OECD Conference on RNAi based pesticides provided an overview on the current status and future possibilities for the regulation of externally applied dsRNA-based products that are proposed for use as pesticides. The event facilitated exchanges between policy makers, academia, industry on their implications in health, environment, and regulation.
Environmental dissipation of dsRNA in soil, aquatic systems and plants - Pame...OECD Environment
10-12 April 2019: The OECD Conference on RNAi based pesticides provided an overview on the current status and future possibilities for the regulation of externally applied dsRNA-based products that are proposed for use as pesticides. The event facilitated exchanges between policy makers, academia, industry on their implications in health, environment, and regulation.
To study of the genetic variations among the Azospirillum lipoferu isolates u...ijsrd.com
Among free-living microorganisms, which can be practically used in agriculture, bacteria from the Azospirillum genus as well as other endophytes are nowadays thought of as the most active component of associative dinitrogen fixation. The investigation was carried out to study the characterization of Azospirillum lipoferu found in the soils of the ten agro-climatic zones which Karnataka, is classified. By using RAPD markers, 75 bands were scored out of which 78.6 % were found to be polymorphic. Statistical analysis of RAPD data enabled the classification of 10 Azospirillum isolates into two major groups. . In this, the cluster analysis based on 75 RAPD bands revealed that the ten A. lipoferu isolates examined clustered at a linkage distance of about 40 units on the dendrogram. There was no correlation between RAPD and geographical origin of isolates.
Assessment of genetic fidelity of in vitro propagated clones of Celastrus pan...iosrjce
Celastrus paniculatus Willd belonging to the family Celastaceae is an endangered Indian medicinal
plant having high pharmaceutical application. The objective of the present investigation was to assess the the
clonal fidelity of in vitro propagated clones of Celastrus paniculatus with the field grown mother plant to
confirm their true to type nature. Micropropagation is an alternative method for the large scale production of
endangered medicinal plants. The genetic stability of in vitro raised clones of celastrus paniculatus were
assessed by using RAPD analysis. Genomic DNA was isolated from healthy and fresh leaves of both mother
plant and in vitro raised plants of Celastrus paniculatus by using CTAB method. Based on the reproducibility of
the primers, 15 RAPD primers were selected for the present investigation. The selected primers gave rise to a
total of 75 scorable bands with an average of 5.1 bands ranging from 300-2700 bp. The number of bands varied
from three (OPQ-07, OPA-13) to seven (OPC-20, OPN-16). Randomly selected 10 micropropagated plants
from each culture period was used. Amplification pattern was electrophoresed in 1.5% TBE, revealing that all
the bands produced by micropropagated plants were monomorphic and similar to that of the field grown plant.
No polymorphism was detected by RAPD analysis.
Transfer of Potential pseudomonas stutzeri genes ROB1323
In this research of saxitoxin bacterial production bioinformatics will be utilized to identify sxt coding genes within pseudomonas stutzeri strain A1501. PCR will then be employed in the isolation of sxt genes from pseudomonas stutzeri. The isolated genes will be transformed into Ecoli cultures.
This presentation gives brief introduction of Recombinant DNA technology. This presentation covers steps involved and tools of Rec DNA Technology. important applications are also explained in this presentation.
This is my dummy proposal for for research design and development course from which I learned a lot about the structure of research proposals and their requirements....
Assessment of genetic fidelity of in vitro propagated clones of Celastrus pan...iosrjce
Celastrus paniculatus Willd belonging to the family Celastaceae is an endangered Indian medicinal
plant having high pharmaceutical application. The objective of the present investigation was to assess the the
clonal fidelity of in vitro propagated clones of Celastrus paniculatus with the field grown mother plant to
confirm their true to type nature. Micropropagation is an alternative method for the large scale production of
endangered medicinal plants. The genetic stability of in vitro raised clones of celastrus paniculatus were
assessed by using RAPD analysis. Genomic DNA was isolated from healthy and fresh leaves of both mother
plant and in vitro raised plants of Celastrus paniculatus by using CTAB method. Based on the reproducibility of
the primers, 15 RAPD primers were selected for the present investigation. The selected primers gave rise to a
total of 75 scorable bands with an average of 5.1 bands ranging from 300-2700 bp. The number of bands varied
from three (OPQ-07, OPA-13) to seven (OPC-20, OPN-16). Randomly selected 10 micropropagated plants
from each culture period was used. Amplification pattern was electrophoresed in 1.5% TBE, revealing that all
the bands produced by micropropagated plants were monomorphic and similar to that of the field grown plant.
No polymorphism was detected by RAPD analysis.
Transfer of Potential pseudomonas stutzeri genes ROB1323
In this research of saxitoxin bacterial production bioinformatics will be utilized to identify sxt coding genes within pseudomonas stutzeri strain A1501. PCR will then be employed in the isolation of sxt genes from pseudomonas stutzeri. The isolated genes will be transformed into Ecoli cultures.
This presentation gives brief introduction of Recombinant DNA technology. This presentation covers steps involved and tools of Rec DNA Technology. important applications are also explained in this presentation.
This is my dummy proposal for for research design and development course from which I learned a lot about the structure of research proposals and their requirements....
Molecular Basis for Genetic Resistance of Fusarium virguliforme, the Causal A...Chloe Siegel
This poster was presented at the Undergraduate Research Symposium at the University of Illinois at Urbana-Champaign. It summarizes a semester-long research project I participated in through the Department of Natural Resources and Environmental Sciences.
This work aimed to (i) Identify and characterize Onion yellow dwarf virus potyvirus (OYDV) in the onion plants in Egypt. (ii) Clone and sequence the coat protein gene of the Egyptian isolate of OYDV and comparing it with other OYDV isolates reported in the GenBank database. (iii) Study the influence of therapeutic doses of kinetin (6-Furfurylaminopurine) on production of virus-free onion plantlets and improve its regeneration ability through in vitro micropropagation.
Summary. Phaeomoniella chlamydospora is an important tracheomycotic fungus involved in different syndromes
of the esca disease complex affecting young and mature grapevine plants. Although grapevine planting material
is frequently infected by this pathogen, no regular screening to detect the fungus is carried out, mainly due to
the lack of fast, sensitive and affordable methods for large-scale application. A monoclonal antibody-based triple
antibody sandwich (TAS) enzyme-linked immunosorbent assay (ELISA) was developed to specifically detect Pa.
chlamydospora from grapevine wood tissues. This assay could be especially useful for large-scale application in
nurseries, to ensure Pa. chlamydospora-free grapevine stocks, and thus contributing to the production of healthy
propagation material.
DOI: 10.14601/Phytopathol_Mediterr-11860
SDS-PAGE electrophoretic analysis of young leaves protein was used to asses the genetic relatedness
among 54 specimens belonging to 6 common cultivars of Phoenix dactylifera L. (Aglany, Amry, Haiany,
Bent Aisha, Samany, and Zaghloul), that were collected from 7 different locations in Ismailia region. A
total of 10 different protein bands were collectively detected in the gels of the 54 studied specimens. The
resulted protein profile comprised one monomorphic band. The maximum number of bands observed was
10 bands found in one specimen of Zaghloul cultivar. While the minimum number of bands observed
was three bands found in one specimen of Aglany cultivar and two specimens of Bent Aisha cultivar. The
data of the allele frequencies of the six studied date palm cultivars revealed that all the alleles in Haiany
cultivar were monomorphic, except in three loci with the lowest percentage of polymorphic loci (30%).
While Bent Aisha and Aglany cultivars have the highest polymorphism (70% and 80%, respectively). In
addition, it was found that the ratio of gene diversity/locus varied greatly within the specimens of the six
studied date palm cultivars. Agglomerative cluster analysis, based on the genetic distances of the studied
54 specimens, revealed the variations and relatedness among the six collected cultivars.
Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggpla...ijtsrd
Eggplant is prone to attack by several pests including bacteria, fungi, nematodes and insects. In this study, we have analyzed phylotype of bacterial wilt Ralstonia solanacearum infection in eggplant plants collected from Bhubaneswar Orissa in India. Bacterial wilt symptomatic five plant samples were collected from brinjal field in Bhubaneswar in 2016. The samples were macerated in sterile distilled water and grown on Kelman's triphenyltetrazolium chloride TZC agar media. Total genomic DNA of the bacterium were extracted and subjected to PCR amplification using the R. solanacearum specific universal primer pair 759 760. An expected single 280 bp fragment amplified in all the samples confirmed the identity of these as Ralstonia. To reconfirmed isolate of bacterium, the amplicon was sequenced in sequencer. In NCBI blast, the nucleotide sequence was 100 similar with Ralstonia solanacearum strain RS lpxC DOB 1 AB910593 and the sequence was submitted in NCBI database under Acc. No. KY393266. To determined phylotype of strain used specific multiplex PCR with phylotype specific primers Nmult 21F1 2, Nmult 22InF, Nmult 23AF, Nmult 22RR revealed that all the five infected samples belonged to phylotype I as a 144 bp amplicon were observed in agarose gel. On the basis of above finding concluded that the bacterial wilt infected eggplant collected from Bhubaneswar was Ralostonia solanacearum, Phylotype I. Rakesh Kumar | Ramachandran, E. | Koteshwar Yadav "Phylotype Analysis of Ralstonia Solanacearum Causing Bacterial wilt in Eggplants in Orissa in India" Published in International Journal of Trend in Scientific Research and Development (ijtsrd), ISSN: 2456-6470, Volume-3 | Issue-3 , April 2019, URL: https://www.ijtsrd.com/papers/ijtsrd21580.pdf
Resistance of some olive (Olea europaea) cultivars and hybrids to leaf spot d...Agriculture Journal IJOEAR
Abstract— In order to investigate the resistance of some olive (Olea europaea L.) cultivars and hybrids to leaf spot disease caused by Venturia oleaginea, this study was conducted on high susceptible cultivar Meski and nine hybrids. Samples were collected from a field site located in Nabeul (North East of Tunisia) and evaluated for their susceptibility to leaf spot disease by means of visible and latent infection. Therefore, the studied plants were classified into three categories: very susceptible, intermediate and resistant. Meski cultivar and three hybrids (MxA) obtained through controlled crosses between Meski and Arbequina were the most susceptible to the disease. The hybrids MxC resulting from the crosses between Meski and Chétoui olive cultivars presented less severity. However, the hybrids obtained through crosses between Meski and Picholine cultivars showed the lowest incidence of infection. Microsatellites were used as markers to analyze the genetic relationships between parental olive cultivars and hybrids and the effects of crossing on the disease resistance. Cluster analyses, using the SSR data, showed that olive cultivars and hybrids obtained by controlled cross between MeskixPicholine, Meski x Arbequina and Meski ×Picholine were related to Picholine cultivar. The hybrid Meski x Chétoui was more related to cultivar Meski. Data analyses revealed that the GAPU101 showed the highest number of alleles (8) followed by the tow loci UDO99 and GAPU71 with 6 alleles. The DCA18 locus showed 5 alleles. Genetic variability was wide as indicated by the values of observed heterozygosity as noted 1.00 at locus of the four studied loci. Polymorphic information content (PIC) varied from 0.669 to 0.776. The gene diversity values were higher than 0.53. Genetic distances were determined based on the SSR genotype data and component principal analysis were used for finding possible correlation between severity disease, Meski cultivar and hybrids.
The present study aims to (I) evaluate the antiviral activity of eugenol oil nanoemulsion (EON) on eliminate Banana bunchy top virus (BBTV) from naturally infected banana plants and produce virus-free banana plants, (II) identify fungal contaminants of in vitro banana cultures and (III) evaluate the potential of EON on the suppression of the identified microbial contaminants and reduce of their occurrence frequency.
Detection of Genetic variation in tissue culture clones of date palm using IS...IJSRD
Date palm is a plant having high nutritional value and long life (yielding up to 100 years). Phoenix dactylifera requires 2-5 males for pollination of 100 females’ plant depending up on genetic and environment factors. Therefore paternity variation expected to very low according to PCR based techniques, Even though we have tried to find out genetic variation among tissue culture cloned plant. Tissue culture technique can be used for genetic improvement of date palm. The main purpose of this study was to evaluate the genetic variation in the tissue culture clones of date palm by using ISSR primers among mother and it’s two clones. The plant DNA was extracted and subjected to detection of genetic variation in two groups of date palm using ISSR primers. In this study ISSR primers produced monomorphic bands within group-1 and group-2. Genetic variation in tissue culture clones of date palm was not detecte by UBC primer series.
Antimicrobial, immunomodulatory and cytotoxic activities of green synthesized...HaloCantik
Calotropis procera and Somra (Acacia) honey are used in traditional medicine. The benefits of mixing 20% Somra honey and C. procera leaf water extract (CPLWExt) were aimed to be studied. Honey/CPLWExt were utilized to produce silver nanoparticles (AgNPs) separately. AgNPs were characterized via UV/Vis and electron microscope scanning. Bio-molecules in CPLWExt/honey were investigated utilizing FT-IR spectroscopy. Biological activities of CPLWExt and honey were tested. The outcomes showed that CPLWExt and honey have numerous functional groups and could produce AgNPs. CPLWExt, CPLWExt + AgNPs, honey and honey + AgNPs hindered the growth of rat splenocytes, while CPLWExt + honey invigorated it. Antimicrobial power was found in CPLWExt and honey, which increased in the presence of AgNPs. Honey/honey + AgNPs suppressed the proliferation of HeLa and HepG2 cells. In conclusion, honey/CPLWExt could produce AgNPs and showed immunomodulatory and antibacterial power. Somra honey/honey + AgNPs have anticancer power. Somra honey + CPLWExt reflected a good immunostimulatory powers that can be nominated as an immunostimulant.
The Effect of Dried Leaves Extract of Hyptis suaveolens on Various Stages of ...iosrjce
IOSR Journal of Pharmacy and Biological Sciences(IOSR-JPBS) is a double blind peer reviewed International Journal that provides rapid publication (within a month) of articles in all areas of Pharmacy and Biological Science. The journal welcomes publications of high quality papers on theoretical developments and practical applications in Pharmacy and Biological Science. Original research papers, state-of-the-art reviews, and high quality technical notes are invited for publications.
II. RNA silencing
RNA silencing/RNAi/ Gene quelling as known in fungi is an endogenous cellular mechanism, that works at the mRNA level and has a sequence dependent mode of action. It is found only in higher eukaryotes and known to be absent in prokaryotes. Initiated by dsRNA or stem loop RNA structures like hairpin RNAs (hpRNAs) which are acted upon by the dicer like enzymes to form 21-25 bp siRNA duplexes, which are further incorporated in the RNA induced silencing complex (RISC) where duplex is unwound and subsequently argonaute 2 protein cleaves the one strand and only the guide strand remains with RISC. Guide strand forms complex with the mRNA through complementary base pairing which is then degraded or silenced through Transcriptional or Post-Transcriptional gene silencing.
III. Transgenic versus non-transgenic RNAi
RNAi in crop protection can be achieved both via transgenic/transformative and non-transgenic/ non-transformative approaches. In transgenic approach, a dsRNA construct has to be genetically engineered into the plant genome by a straightforward strategy of transformation and it provides efficient and long term control. But it also requires generations of crop plants, taking years and delaying practical application also due to the extensive regulatory procedures. As well as during genetic engineering plant transformability and genetic stability needs to be taken into account. Especially, the main issue is that there is very low consumer acceptance of genetically modified crops.
Whereas in case of non-transformative RNAi, dsRNAs are topically applied on the plants either as spray suspension, trunk/stem/petiole injection, root absorption or seed treatment, etc. As this approach silence the genes without introducing any heritable changes in the plant genome, they may not be regulated as a genetically modified crop thus excluding the issue of consumer acceptance. Moreover they are feasible and affordable and incur less cost per tree. The only issue may lie with the continuous supply of dsRNAs which may be overcome using nanoparticles like clay nanosheets, carbon nanotubes, cationic nanoparticles or chemicals like surfactants or peptide based RNA delivery system can be used while not only improves the stability and absorption of dsRNA but also provide for the slow and sustained release of them.
One such study has been done by Mitter et al., wherein they sprayed the dsRNA suspension on the plants either singly or after loading them on clay nanosheets followed by inoculation of the challenge virus. They observed that the clay nanosheets provided enhanced stability and sustained release of dsRNA increasing the protection window from 5 to 20 days in a single spray. This also confirms that the dsRNAs could move or translocate to untreated parts of the plant providing systemic protection to the plants.
IV. Mode of entry and movement of dsRNAs in plants
The dsRNAs sprayed on the plants penetrate the leaves through the wounds o
The First Workshop of Plant Pathology Research Institute
The Modern Trends in Controlling Plant Diseases
10-11 February 2019
Under auspices of
Prof. Dr. Eszzaldin Omar Abusteit
Minster of Agricultural and Land Reclamation
Prof. Dr. Mohamed Soliman
Director of Agricultural Research Center
&
President of the workshop
Prof. Dr. Ashraf El Saied Khalil
Director of Plant Pathology Research Institute
The objective of this study was to examine the antiviral activity of native lactoferrin against Potato virus x, the most important virus that severely affects potato crop and productivity in Egypt, using tissue culture technique and spraying the plants in greenhouse by the aqueous solution of lactoferrin.
Research topic was come from successful inactivation of some plant viruses by gamma irradiation like Citrus tristeza virus, Necrotic ring spot virus and Prune dwarf virus. Gamma irradiation has been also used to sterilize agricultural products in order to increase their conservation time or to reduce pathogen when being traded from a country to another. Gamma radiation is high-energy radiation emitted from certain radioactive isotopes as cobalt 60, these isotopes are potential sources of gamma radiation. Therefore, this research was conducted to find out the inactivation possibility of Hibiscus witches' broom (HibWB)-phytoplasma using gamma irradiation through tissue culture technique with clarify their effect on in vitro growth and survival rate.
The present study has been conducted to perform the following objectives:
1- Study the effect of different temperature degrees and meristem culture technique on elimination of Potato leafroll virus (PLRV) and Potato virus x (PVX), from the most commonly potato cultivars in Egypt (Spunta and Lady Rosette).
2- Production of potato minitubers from direct transplanting of in vitro virus-free plantlets in greenhouse. Also, investigate the effect of different soil mixtures on minitubers production for both cultivars.
The objectives of this study were to detect and characterize the phytoplasma in tissues of diseased hibiscus plants using Dains’ stain light microscopy and molecular based techniques. Molecular characterization was performed using the DNA sequencing and phylogenetic analysis of the spacer region between 16S and 23S rRNA fragment of the isolated phytoplasma genome. This work concerning phytoplasma associated witches' broom (group 16SrII) diseases of hibiscus plants is achieved for the first time in Egypt.
The objectives of this study to investigate the occurrence and etiology of tomato malformation recently observed in Egypt and illustrate the responsibility of associated phytoplasma. Also, the study was extended to characterize associated phytoplasma, based on molecular techniques.
The initial objective of this work was to isolate Tomato spotted wilt virus (TSWV) from asymptomatic infected plants and then identify the virus on the basis of biological properties among the most common or other hosts if possible after inoculation. Phylogenetic analysis was then conducted to gain information about the similar identity of the TSWV-isolate that reported in this study with available TSWV sequences from other parts of the world.
The objectives of this study are: (i): To investigate and recognize the internal and abnormalities impacts induced by phytoplasma infection in the tomato host according to recent studies have shown that the association between plants and phytoplasmas can result in anatomical alteration in phloem tissues of infected plants, and great differences between healthy and diseased samples using microscopic examination of longitudinal, cross or ultra-thin sections of leaf blade, leaf petiole and stem. (ii): To determine the efficiency of different techniques toward production of phytoplasma-free tomato plantlets and mitigation of phytoplasma disease.
زراعة الأنسجة النباتية
طرق الإكثار بزراعة الأنسجة النباتية
البيئة الغذائية المستخدمة فى الزراعة
مراحل زراعة الأنسجة النباتية
العوامل المؤثرة فى نجاح زراعة الأنسجة
العقبات التى تنشأ أثناء مراحل زراعة الأنسجة النباتية
مجالات زراعة الأنسجة النباتية
بدأت تقنية زراعة أنسجة النبات بهدف اكثار وإنتاج نباتات خالية من الأمراض النباتية، وتعددت أشكال هذه التقنية مثل زراعة القمم المرستيمية للنبات، وإستخدام العلاج الحرارى أو الكيماوي، وقد واكب هذا ظهور طرق حديثة فعالة تمثل مصادر جديدة للتغايرالوراثى فى برامج التربية وتحسين النبات وتعتبر من وسائل المقاومة الحديثة التى تعرف باسم المقاومة المستحثة وذلك عن طريق تحفيز أو حث جزء من أجزاء النبات لدفعه على مقاومة الأمراض بإستخدام وسائل متنوعه مثل الحث الفيزيائى عن طريق إستخدام الأشعه فوق البنفسيجية أوأشعة جاما.
ومن هنا جاء إستخدام هذه التقنية الجديدة في مجال زراعة الأنسجة النباتية والتي تعتمد على حث البراعم الداخلية على التوالد والتكاثر بدون تكوين الكالس وإنتاج شتلات خالية من الأمراض النباتية مع تجانس النباتات فى النمو والحصول على أعداد كبيرة من النباتات فى أقل وقت، حيث أمكن تطبيقها بنجاح عن طريق إستخدام أشعة جاما ونقلها لمحاصيل مختلفة كالطماطم (Solanum lycopersicon L.) والهيبسكس (Rosa-sinensis L.) للمرة الأولى فى مصر لمقاومة أمراض الفيتوبلازما المختلفة وتقييم جرعات مختلفة من أشعة جاما لتحديد الجرعات المناسبة لكل نبات وكذلك إختيار الأوساط الغذائية المناسبة لزراعة الأجزاء النباتية المشععة وذلك فى معمل زراعة الانسجة بقسم بحوث الفيروس والفيتوبلازما بمعهد بحوث أمراض النباتات – مركز البحوث الزراعية بالجيزة.
أن تطبيق أدوات جديدة تعتمد على تقنية زراعة الأنسجة والتشعيع باستخدام أشعة جاما قد تساعد على إقتراح استراتيجيات فعالة لمقاومة الأمراض النباتية بصفة عامة وأمراض الفيتوبلازما بصفة خاصة وكذلك منع إنتشارها بالإضافة إلى تحقيق التنمية الزراعية ودفع عجلة النمو والتقدم الزراعى عالمياً وزيادة الصادرات عن طريق تحسين إنتاجية المحاصيل المختلفة وزيادة كمية الغذاء التى ينتجها النبات الواحد وتقليل الفقد فى المحصول نتيجة الإصابة بالأمراض وذلك للايفاء بالمتطلبات المتزايدة للبشرية نتيجة الزيادة الهائلة فى أعداد السكان وتقليل معدلات الإستيراد واخيراً تحقيق الاكتفاء الذاتى.
Deep Behavioral Phenotyping in Systems Neuroscience for Functional Atlasing a...Ana Luísa Pinho
Functional Magnetic Resonance Imaging (fMRI) provides means to characterize brain activations in response to behavior. However, cognitive neuroscience has been limited to group-level effects referring to the performance of specific tasks. To obtain the functional profile of elementary cognitive mechanisms, the combination of brain responses to many tasks is required. Yet, to date, both structural atlases and parcellation-based activations do not fully account for cognitive function and still present several limitations. Further, they do not adapt overall to individual characteristics. In this talk, I will give an account of deep-behavioral phenotyping strategies, namely data-driven methods in large task-fMRI datasets, to optimize functional brain-data collection and improve inference of effects-of-interest related to mental processes. Key to this approach is the employment of fast multi-functional paradigms rich on features that can be well parametrized and, consequently, facilitate the creation of psycho-physiological constructs to be modelled with imaging data. Particular emphasis will be given to music stimuli when studying high-order cognitive mechanisms, due to their ecological nature and quality to enable complex behavior compounded by discrete entities. I will also discuss how deep-behavioral phenotyping and individualized models applied to neuroimaging data can better account for the subject-specific organization of domain-general cognitive systems in the human brain. Finally, the accumulation of functional brain signatures brings the possibility to clarify relationships among tasks and create a univocal link between brain systems and mental functions through: (1) the development of ontologies proposing an organization of cognitive processes; and (2) brain-network taxonomies describing functional specialization. To this end, tools to improve commensurability in cognitive science are necessary, such as public repositories, ontology-based platforms and automated meta-analysis tools. I will thus discuss some brain-atlasing resources currently under development, and their applicability in cognitive as well as clinical neuroscience.
Nutraceutical market, scope and growth: Herbal drug technologyLokesh Patil
As consumer awareness of health and wellness rises, the nutraceutical market—which includes goods like functional meals, drinks, and dietary supplements that provide health advantages beyond basic nutrition—is growing significantly. As healthcare expenses rise, the population ages, and people want natural and preventative health solutions more and more, this industry is increasing quickly. Further driving market expansion are product formulation innovations and the use of cutting-edge technology for customized nutrition. With its worldwide reach, the nutraceutical industry is expected to keep growing and provide significant chances for research and investment in a number of categories, including vitamins, minerals, probiotics, and herbal supplements.
Slide 1: Title Slide
Extrachromosomal Inheritance
Slide 2: Introduction to Extrachromosomal Inheritance
Definition: Extrachromosomal inheritance refers to the transmission of genetic material that is not found within the nucleus.
Key Components: Involves genes located in mitochondria, chloroplasts, and plasmids.
Slide 3: Mitochondrial Inheritance
Mitochondria: Organelles responsible for energy production.
Mitochondrial DNA (mtDNA): Circular DNA molecule found in mitochondria.
Inheritance Pattern: Maternally inherited, meaning it is passed from mothers to all their offspring.
Diseases: Examples include Leber’s hereditary optic neuropathy (LHON) and mitochondrial myopathy.
Slide 4: Chloroplast Inheritance
Chloroplasts: Organelles responsible for photosynthesis in plants.
Chloroplast DNA (cpDNA): Circular DNA molecule found in chloroplasts.
Inheritance Pattern: Often maternally inherited in most plants, but can vary in some species.
Examples: Variegation in plants, where leaf color patterns are determined by chloroplast DNA.
Slide 5: Plasmid Inheritance
Plasmids: Small, circular DNA molecules found in bacteria and some eukaryotes.
Features: Can carry antibiotic resistance genes and can be transferred between cells through processes like conjugation.
Significance: Important in biotechnology for gene cloning and genetic engineering.
Slide 6: Mechanisms of Extrachromosomal Inheritance
Non-Mendelian Patterns: Do not follow Mendel’s laws of inheritance.
Cytoplasmic Segregation: During cell division, organelles like mitochondria and chloroplasts are randomly distributed to daughter cells.
Heteroplasmy: Presence of more than one type of organellar genome within a cell, leading to variation in expression.
Slide 7: Examples of Extrachromosomal Inheritance
Four O’clock Plant (Mirabilis jalapa): Shows variegated leaves due to different cpDNA in leaf cells.
Petite Mutants in Yeast: Result from mutations in mitochondrial DNA affecting respiration.
Slide 8: Importance of Extrachromosomal Inheritance
Evolution: Provides insight into the evolution of eukaryotic cells.
Medicine: Understanding mitochondrial inheritance helps in diagnosing and treating mitochondrial diseases.
Agriculture: Chloroplast inheritance can be used in plant breeding and genetic modification.
Slide 9: Recent Research and Advances
Gene Editing: Techniques like CRISPR-Cas9 are being used to edit mitochondrial and chloroplast DNA.
Therapies: Development of mitochondrial replacement therapy (MRT) for preventing mitochondrial diseases.
Slide 10: Conclusion
Summary: Extrachromosomal inheritance involves the transmission of genetic material outside the nucleus and plays a crucial role in genetics, medicine, and biotechnology.
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Slide 11: Questions and Discussion
Invite Audience: Open the floor for any questions or further discussion on the topic.
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Observation of Io’s Resurfacing via Plume Deposition Using Ground-based Adapt...Sérgio Sacani
Since volcanic activity was first discovered on Io from Voyager images in 1979, changes
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PARTIAL CHARACTERIZATION OF PRUNUS NECROTIC RINGSPOT VIRUS ON APPLE IN EGYPT
1. Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
Partial characterization of Prunus Necrotic Ringspot Virus on apple
in EGYPT
Aly M. Abdel-Salam1
and Samah A. Mokbel2
1
Plant Pathology Department, Faculty of Agriculture, Cairo University, Giza 12613, Egypt.
2
Virus and Phytoplasma Research Department, Plant Pathology Research Institute, Agricultural Research Center
(ARC), Giza 12619, Egypt.
ABSTRACT
A severe isolate of Prunus necrotic ringspot virus (PNRSV) was isolated from apple
orchards in the vicinity of Nubaria city, Beheira governorate, Egypt. Infected-apple
trees showed chlorotic, necrotic ringspots, and shoot holes on leaves. Severely
infected- trees withered, became useless, and were removed causing severe economic
losses. Reverse transcriptase (RT) polymerase chain reaction (PCR), RT-PCR, using
degenerate primer pair for the coat protein (CP) gene of Ilarvirus amplified products
similar to those produced from peach and apricot isolates of PNRSV-infecting stone
fruits. Dot blotting immuno-binding assay (DBIA) showed positive reaction between
PNRSV-infected apple sap and an Egyptian antiserum for PNRSV. Purified
preparation from infected leaves, using the electro-elution technique yielded
nucleoprotein which had Amax and Amin at 260 and 240 nm respectively. Electron
microscopy examination showed spherical virions with ca. 26 nm in diameter.
Key words: Prunus necrotic ringspot virus, Ilarvirus, RT-PCR, Dot blotting immuno-binding
assay, Egypt
INTRODUCTION
PNRSV belongs to the family
Bromoviridae, genus Ilarvirus
(isometric labile ringspot viruses)
(Fulton, 1983) and includes many
strains that differ in pathogenicity
(Howell and Mink, 1988), biophysical
(Crosslin and Mink, 1992) and
serological properties (Spiegel et al.,
1999). All genera of Bromoviridae
including Ilarvirus contain tripartite
genomes. The RNA1 and RNA2 code
for proteins involved in viral
replication and the RNA3 codes for
both a movement protein and the viral
coat protein (Murphy et al., 1995).
These species of RNAs are
encapsulated in isometric particles (23-
27 nm in diameter) rounded in profile
and without a conspicuous capsomere
arrangement (Brunt et al, 1996).
PNRSV is graft, pollen and seed-
transmitted (Gella, 1980, Uyemoto et
al., 1992; Amari et al., 2004). The
virus in most hosts induces shock
symptoms after infecting plants,
provided that they have not been
infected earlier by latent strains of
PNRSV.
PNRSV is the most common virus
infecting Prunus species as peach and
apricot (Mink, 1992, Myrata et al.,
2003; Abdel-Salam et al., 2008a),
rosaceous plants (Abdel-salam et al.,
2008b), and naturally infecting other
non-rosaceous plants (Abdel-Salam et
al., 2006a). PNRSV is responsible for
yield losses of up to 15% in sweet
cherry and up to 100% in peach.
PNRSV can reduce bud development
in nurseries, decrease growth of fruit
(10% to 30%) and fruit yield (20% to
60%), delay fruit maturity, and
increase susceptibility to winter
injuries in orchards (Oliver et al.,
2009; Pallas et al., 2012).
In Egypt isolates of PNRSV were
detected in peach and apricot grooves
2. Aly M. Abdel-Salam and Samah A. Mokbel
Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
(Abdel-Salam et al. 2008a), Rosa spp.
(Abdel-Salam et al., 2008b) as well as
on sugarbeet plantations (Abdel-Salam
et al., 2006a). In the present study,
incidence of PNRSV is reported on
apple (Malus domestica). Severe
symptoms mimic infections with PNRSV
were recently detected on apple from
several orchards in the vicinity of Nubaria
city, Beheira governorate. Infected-apple
samples were brought to the laboratory
for further detection at serological and
molecular levels to check the presence
of virus. The present study reports the
presence of an isolate of PNRSV on
apple, viz. PNRSV-Apple.
MATERIALS AND METHODS
Virus isolates
An isolate of PNRSV was isolated
from apple groves, Beheira
governorate, Egypt. Infected samples
showed chlorotic, necrotic ringspot,
and shot holes. The virus isolate was
purified biologically by mechanical
inoculation on Chenopodium quinoa
and Gomphrena globsa as described by
Abdel–Salam et al. (1997, 2006a).
Isolates for PNRSV from apple and
apricot, preserved in the greenhouse
facilities, Cairo University were used
as positive controls.
Serologic studies
Dot blotting immunobinding
assay (DBIA) test, described by
(Abdel-Salam et al., 2014) was used in
measuring virus presence in tested
hosts and serologic relationships
between PNRSV isolates from apple,
peach and apricot. Tissue samples were
ground, filtered and diluted 1/10 in
PBST buffer. An Egyptian antiserum
for PNRSV prepared for the peach
isolate (Abdel-Salam et al., 2008a) of
the virus was used in the present study.
Virus purification
Fifty grams of fresh tissues of
PNRSV-infected gomphrena
(Gomphrena globosa) plants,
previously inoculated with PNRSV-
Apple, were used in virus purification.
Purification of PNRSV-Apple utilized
the electro-elution (EE) technique
described by Abdel-Salam (1999).
The EE technique involved
extraction of tissues (1:3 w/v) in 0.1 M
NaH2PO4-Na2HPO4, pH 7.0,
containing 1 mM EDTA, 20 mM
Na2SO3, and 0.1% of each of 2-
mercaptoethanol and thioglycolic acid.
The extract was clarified with 12.5%
volume of each of chloroform and
butanol. The clarified-virus suspension
was concentrated with 4%
polyethylene glycol (4000, mw) and
1% NaCl. The concentrated virions
were suspended in 1 mM phosphate
buffer, pH 7.2, containing 1mM EDTA
(suspension buffer, SB). The virions
were further purified with EE-ISCO
tank with tank buffer containing 20
mM phosphate buffer, pH 7.2, and
applying 4 mA/cell. The concentrated
virions were then suspended in SB and
measured spectrophoto-metrically.
Electron microscopy
Purified virus isolates were stained
with 2% phosphotungestic acid, pH 7.2
according to Fulton (1981).
.
Genomic studies
Extraction of total RNA
Total RNA was extracted from
PNRSV-infected peach and apricot
plants by applying the silica-based
technique described by Boom et al.
(1990).
RT-PCR
The primer set CP (+) sense primer
(5' CCG AAT TTG CAA TCA TAC
CCA CGC T 3') and CP (-) antisense
primer (5' CGG AGA AAT TCG AGT
GTG C 3') complementary to the
3. Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT
Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
conserved region of the coat protein
(CP)gene were used to generate 704 bp
fragments from PNRSV Cp gene
(RNA-3) as described by Abdel-Salam
et al.(2008).
First strand cDNA was synthesized in a
total of 20 μl reaction mixture. 5 μl of
total RNA (~25 μg) was heated at 65o
C
for 8 min, chilled for 3 min in ice, then
added to 15 μl reaction mixture
containing 1.5 μl of antisense primer
(10 pmol), 2 μl of M-MulV reverse
transcriptase buffer (10X), 0.125 μl of
M-MulV reverse transcriptase (5000
U), SibEnzyme Ltd, 2 μl dithiothreitol
(100 mM), Promegam, 1 μl dNTPs
mix (10mM), 0.25 μl of ribonuclease
inhibitor (40 U/ μl), Promega, and
8.125 μl DEPC H2O. The mixture was
incubated at 42o
C for 1 h, incubated at
95o
C for 3 min, and then kept in ice.
PCR cocktail included 2 μl of the
reverse transcription products, 5 μl of
5X Green GoTaq buffer, containing
1.5 mM Mg2Cl, Promega, 0.5 μl of
dNTPs mix (10mM), 1 μl of each of
sense and antisense CP primers (10
pmol, each), 0.25 μl of Go Taq
polymerase (5 U/ μl), Promega, 12.25
μl DEPC H2O. PCR conditions
included 5 min at 95o
C; followed by 35
cycles of amplification of 1 min at
95o
C, 1 min at 53o
C, 1 min at 72o
C,
and held for 10 min at 72o
C. The RT-
PCR amplicons were analyzed on 1 %
agarose at 100 V in 1/2 X TAE (40
mM Tris/acetate, 1 mM EDTA, pH
8.0) and stained with ethidium bromide
and examined with UV trans-
illuminator.
RESULTS
Smptomatology
Primary symptoms on apple starts
with leaf-chlorotic ringsspot which
turned necrotic afterwards (Fig.1-A).
Shot hole symptoms followed the
necrotic ringspot formation (Fig.1-B).
Few small apple fruits are formed on
infected tree (Fig. 1-C). Infected trees
express shock symptoms (Fig.1-D)
which extend to circumvent the whole
tree and causing leaf defoliation (Fig.
1-E). Severely infected- trees wither out
and carry no fruits (Fig. 1-E).
Serologic study
DBIA test was used to detect
PNRSV-Apple isolate. Results in
Figure (2) showed the positive reaction
of sap from infected apple, apricot and
peach with the induced antiserum for
PNRSV-Peach isolate. The peach
isolate of PNRSV reacted strongly
with its homologous antiserum. While
both isolates of apple and apricot of
PNRSV reacted moderately with the
antiserum of PNRSV-Peach indicating
distant serologic relationship with the
peach isolate of PNRSV.
Virus purification
The purified virus preparations,
using EE technique, for PNRSV-
Apple, had a UV spectrum typical to
nucleoproteins with Amax at 260 nm,
Amin at 240 nm, with A260/280 ratio of
1.6 (Fig. 3). Such results are typical to
similar values reported for different
isolates of PNRSV).
(e.g PNRSV (Abdel-Salam et al.,
2006a, 2008a, b), some begomoviruses
(Abdel-Salam, 1999, Abdel-Salam et
al 2006b), an ipomovirus, and a
crinivirus (Abdel-Salam, 2012). The
EE technique is fast, low cost, and
meets the demands of many
moderately equipped laboratories.
Electron microscopy examination
Electron microscopy results of
purified PNRSV showed spherical
virions with an average diameter is 26
nm (Fig. 4).
4. Aly M. Abdel-Salam and Samah A. Mokbel
Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
5. Partial characterization of Prunus Necrotic Ringspot Virus on apple in EGYPT
Molecular studies
RT-PCR detection of PNRSV from
apple, peach and apricot
RT-PCR successfully detected
PNRSV-viral RNA from apple, peach,
and apricot tissues (Fig.5). A full
length CP gene DNA fragment about
704 bp in size was detected from the
three tested isolates of PNRSV using
specific primers for PNRSV- CP gene
(Fig.5). No signal was detected in the
negative control.
DISCUSSION
Characterization of the present
isolate of PNRSV was based on
symptomatology, chemical and
physical properties, serology,
molecular analysis and electron
microscopy.
Symptomatology
The described symptoms on
infected apple trees are similar to
symptoms caused by PNRSV infection
on several stone fruits (Pusey. and
Yadava, 1991; Mink, 1992; Abdel-
Salam et al. 2008a).
Serologic study
The tested PNRSV isolates from
peach, apricot, and apple reacted
serologically with a local antiserum
prepared for PNRSV-peach isolate. As
expected intensity of the reaction was
correlated with degree of homology
between the antiserum and its
respective isolate; being strong with
the homologous peach isolate and
moderate with the heterologous apricot
and apple isolates. Such results agree
with results of Crosslin and Mink
(1992), Spiegel et al. (1999), and
Abdel-Salam et al. (2006a) who
reported the serologic diversity
between PNRSV isolates.
Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
6. Aly M. Abdel-Salam and Samah A. Mokbel
Egyptian J. Virol, Vol. 11 (2): 280-287, 2014
Virus purification
The purified virus preparations had
physical characters typical to
nucleoproteins obtained for several
isolates of PNRSV (Fulton, 1981,
Crosslin and Mink, 1992; Abdel-Salam
et al., 2006a, 2008a, b). The EE
purification method used in the present
study was also successful in purifying
other ilarviruses (e.g PNRSV (Abdel-
Salam et al., 2006a, 2008a, b), some
begomoviruses (Abdel-Salam, 1999,
Abdel-Salam et al 2006b), an
ipomovirus, and a crinivirus (Abdel-
Salam, 2012). The EE technique is
fast, low cost, and meets the demands
of many moderately equipped
laboratories.
Electron microscopy examination
Purified virions had an average
diameter of 26 nm thus resembling
members of Ilarvirus as reported by
(Brunt et al, 1996) and Abdel-Salam et
al (2006a, 2008a, b).
Molecular studies
Specific primers for the CP gene
of PNRSV amplified the full length CP
from PNRSV-infected apple, peach
and apricot; confirming therefore the
presence of PNRSV in the tested
isolates. Similar results were obtained
by other authors using RT-PCR for the
detection of PNRSV in stone fruits
(Aparicio et al., 1999; Moury et al.,
2001; Ulubas and Ertunc, 2004).
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