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Home > Biological and Molecular Characterization of Begomoviruses from Pothwar Region of Pakistan

Biological and Molecular Characterization of Begomoviruses from Pothwar Region of Pakistan

Thesis Info

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Author

Riaz, Hasan

Program

PhD

Institute

Pir Mehr Ali Shah Arid Agriculture University

City

Rawalpindi

Province

Punjab

Country

Pakistan

Thesis Completing Year

2016

Thesis Completion Status

Completed

Subject

Plant Pathology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9709/1/Complete%20Hasan%20Riaz%20PhD%20Thesis.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676725621942

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Begomoviruses (family Geminiviridae; genus Begomovirus) are dicot plantsinfecting viruses which are exclusively transmitted by the insect vector whitefly (Bemesia tabaci). These viruses belong to economically important group of viruses which have monopartite and bipartite single stranded DNA genome. The plant diseases induced by begomoviruses are major constraints for successful crop production worldwide. Pothwar region is located in the northern part of Punjab province with an estimated area of 22, 254 km2. Based on the ecological and climatic conditions Pothwar region is adapted for the production of large number of agricultural and horticultural crops. The lack of systematic study on begomoviruses diversity in Pothwar region gave a strong backing to current study for assessing the incidence, distribution and genetic characterization of begomoviruses in the region. A total of 138 plant samples belonging to 17 different plant families were collected from four districts of Pothwar region and capital territory Islamabad. Total genomic DNA was isolated from all plant samples by CTAB method followed by PCR employing universal degenerate primers for begomoviruses detection. Selected positive samples were subjected to Rolling Circle Amplification (RCA) for full genome amplification of DNA-A, DNA-B and satellite genomes which were cloned and sequenced subsequently. The obtained sequences were analyzed using Sequence Demarcation Tool (SDT) and phylogenetic analysis was carried out by Molecular Evolutionary Genetics Analysis (MEGA) 6.0. Plants belonging to 15 families were tested positive for begomoviruses and the overall incidence of begomoviruses in Pothwar region was xxii 77.5 percent while incidence in individual areas was 85.7 percent in Attock, 85 percent (Rawalpindi), 83.3 percent (Islamabad), 81 percent (Chakwal) and 55.5 percent (Jhelum). The begomoviruses were not evenly distributed in Pothwar region with 32 percent in Rawalpindi, 27 percent (Chakwal), 22.4 percent (Attock), 14 percent (Islamabad) and 05 percent (Jhelum). The most significant contribution of the present study is the identification and characterization of new begomovirus specie which was tentatively named as “Euphorbia yellow leaf curl virus” (EYLCV) [Pakistan:Rawalpindi:Euphorbia:2012] [KM978186]. The EYLCV was also isolated and characterized from Hibiscus syriacas and Combretum indicum. Other begomoviruses which were characterized in present study are Mesta yellow vein mosaic virus (MeYVMV) and its association with Cotton leaf curl Multan Betasatellite (CLCuMB) infecting Malvastrum coromandelianum, Rose leaf curl virus (RoLCuV) infecting Duranta repens, Tomato leaf curl New Dehli Virus infecting Solanum lycopersicum and Chilli leaf curl virus and its associated betasatellite infecting Capsicum annum from Pothwar region. The Pothwar region provides an opportunity for helper viruses to develop more complex relationships with satellites DNAs altogether opening a window for the development of unique begomoviruses disease complexes with likely implication of invading the cotton belt that could result in severe economic losses to national economy.
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بے سہاروں کا یہاں بوجھ اُٹھانے کے لیے

بے سہاروں کا یہاں بوجھ اُٹھانے کے لیے
کوئی تیار نہیں اپنا بنانے کے لیے

پیار ہر ایک سے کرنا ہی مری دعوت ہے
میرا پیغامِ محبت ہے زمانے کے لیے

کیا ہوا تجھ کو ترا یار اگر چھوڑ گیا
میں جو حاضر ہوں ترے ناز اُٹھانے کے لیے

میں نے جس شخص کا ہر وقت بھلا سوچا ہے
وہ ہے بے تاب مری خاک اُڑانے کے لیے

گو کہ مشکل ہے کہ تائبؔ جی سکوں حاصل ہو
ہم تو زندہ ہیں فقط رنج اُٹھانے کے لیے

Advancing Age as a Risk Factor for Acute Myocardial Infarction

Background: Acute myocardial infarction (AMI) is one of the leading causes of death in developed and developing countries. Age is an important non-modifiable risk factor for acute myocardial infarction. Objectives: The objective of the study was to explore the relationship of advancing age with the risk of acute myocardial infarction. Methods: It was a cross-sectional study conducted in 2019 after getting approval from Institutional Review board of University of Health Sciences, Lahore. Written informed consent and thorough history was taken from the study participants. Group 1 included 45 AMI patients aged 20-60 years. Group 2 included 45 healthy individuals aged 20-60 years. Independent sample t test and chi-square tests were applied for analysis of data. Results: Mean age was significantly higher in AMI patients (50.52±7.31) as compared to healthy controls (30.67±7.20). The risk of AMI increases with advancing age (p<0.001, OR= 2.78). Conclusions: Advancing age is an important risk factor for acute myocardial infarction.

Molecular Identification and Characterization of Some Novel Halotolerant Rhizobacteria Containing Acc-Deaminase, for Growth Promotion of Phaseolus Vulgaris L. under Salt Stress

Soil salinity is one of the factors that most severely affect plant growth and production around the world. The rhizosphere supports the development and activity of a huge and diversified microbial community, including microorganisms capable to promote plant growth. Bacterial strains have variable ability to tolerate the salt stress and some salt tolerant rhizobium strains can grow at NaCl concentration up to 500 mM. Such bacterial strains are halophiles, with distinctive physiological structure and hereditary properties, salt-tolerant bacteria go through some morphological, metabolic and structural modifications to tolerate salt stress. These halo tolerant rhizobacteria colonize roots of monocots and dicots, and enhance plant growth by direct and indirect mechanisms. Modification of root system architecture by plant growth promoting bacteria implicates the production of phytohormones and other signals that lead, mostly, to enhanced lateral root branching and development of root hairs. Plant growth promoting bacteria also modify root functioning, improve plant nutrition and influence the physiology of the whole plant.Such halophilic microorganisms have captivated the attention of scholars at home and abroad. Our country has large salt deposits providing suitable condition for growth of salt loving microbes. Such salt deposits are best source for isolation of halo tolerant bacteria. In this project plants were collected from Karak salt mines (Bahadur Khel, Jatta Ismail Khel) and Khewra salt mines of Pakistan. The salt deposits of Khewra and Karak are enormously saline and the microbial communities in such areas have not been explored yet. This study reports isolation, identification and characterization of halo tolerant bacteria. This report also represents novel bacteria that require NaCl for its growth. Total sixty four bacterial strains were isolated from the rhizosphere of plants collected from Karak salt mines (Bahadur Khel, Jatta Ismail Khel) and Khewra salt mines of Pakistan belonging to fourteen different genera. Such bacterial strains can grow in media with different salt concentrations. Morphological and biochemical characteristics of these strains were studied by optimizing their growth conditions such as pH range 5-10 and NaCl range 0-30%. High microbial growth was observed at low salt concentration. Bacterial strains were identified on the basis of 16S rRNA gene sequence and phylogenetic analysis demonstrated that these are closely related to species belonging to different genera: Halomonas, Kushneria, Alcanivorax, Brevibacterium, Planococcus, Staphylococcus, Zhihengluella, Bacillus, Tenuibacillus, Thalassobacillus, Salinicola, Brachybacterium, Piscibacillus and Halobacillus. NCCP-934 is candidate novel strain in this study. Based upon phylogenetic analysis, physiological and chemotaxonomic data and DNA-DNA hybridization values, strain NCCP-934T belong to the genus Kushneria and showed the highest similarity as 98.9% with Kushneria marisflavi SW32T (AF251143), 98.7% with K. indalinina CG2.1T (AJ427627), 98.4% with K. avicenniae MW2aT (DQ888315) and less than 95% with the other species of the genus Kushneria and other taxa of the related genera. Strain NCCP-934T was characterized as Kushneria pakistanensis. Cells of strain NCCP-934T are Gram-stain negative, motile, non-spore forming rods and strictly aerobic. The optimal growth conditions occur at 30–33ºC and a pH of 7.0–9.0. Strain NCCP-934T tolerated to 30% NaCl with optimum growth at 3–9% (w/v) NaCl and thus, it can be considered as a moderately halophilic microorganism. DNA-DNA relatedness between strain NCCP-934T and other related strain was less than 30%. The data supported the affiliation of NCCP-934T with genus Kushneria. On the basis of results strain NCCP-934T distinguished from closely related strains and represented a novel species in the genus Kushneria, for which the name Kushneria pakistanensis sp. nov. is proposed with the type strain, NCCP-934T (=LMG 28525T = KCTC 42082T = JCM 18802T) is AB970675. Isolated bacterial strains and novel strain were characterized for their plant growth promoting properties. For this purpose PCR amplification of nifH and acdS genes was done. Bacterial strains with nifH gene and acdS genes were inoculated to Phaseolus vulgaris under sodium chloride stress. Inoculated bacterial strains showed positive results indicating that such bacterial strains can be utilized for growth promotion of different plants under salt stress.