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Use of a Plant Virus Vector for Screening of Gene Constructs With Potential Insecticidal Activity

Thesis Info

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Author

Shaheen, Musarrat

Program

PhD

Institute

Pakistan Institute of Engineering and Applied Sciences

City

Islamabad

Province

Islamabad.

Country

Pakistan

Thesis Completing Year

2017

Thesis Completion Status

Completed

Subject

Biotechnology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/13337/1/Musarrat%20Thesis%2014-7-2017.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727643008

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Extending the novel implications of plant viruses as invaluable genetic tools in the field of transgenic plant technology, Potato Virus X (PVX) based transient gene expression is a simple rapid, inexpensive methodology employed for preliminary screening and evaluation of insecticidal gene constructs in plant sucking insect species. In this study, PVX vector was employed to achieve high throughput expression of two-candidate insecticidal proteins viz. Hvt, (Hydronyche versuta spider) and ACA Lectin (Allium cepa agglutinin) in tobacco plants. The Reverse transcriptase (RT-PCR) revealed a high throughput expression of 117 bp Hvt, 325 bp ACA Lectin in tobacco were bioassayed in Phenacoccus solenopsis Tinsley (P. solenopsis Tinsley), Aphis gossypii Glover, (A. gossypii Glover), M. persicae Sulzer (M. persicae Sulzer) and herbivorous insect species of Helicoverpa virescens (H. virescens), Spodoptera littoralis (S. littoralis). The results revealed severe insecticidal and antagonistic effect of Hvt in terms of significant insect mortality p<0.01%, reduced population survival and nymph production in P. solenopsis Tinsley, A. gossypii Glover and M. persicae Sulzer. ACA Lectin also exerted significant anti-feedent and insecticidal characteristics at p<0.01% on colonization and population survival in tested insects and larval mortality and lower dry weight gain in herbivorous insect species of H. virescens and S. littoralis. The practical application of PVX vector was extended through PVX- based VIGS mediated production of dsRNAs of two partial insect gene transcripts i.e. Voltage Gated Calcium Channel (PSCaVα1) and Elongation Factor (PSEF-1α) in tobacco plants and RNAi response in P. solenopsis Tinsley. The Reverse Transcriptase (RT-PCR) revealed a high throughput expression of 117 bp Hvt, 325 bp ACA Lectin and plants dsRNAs of 152 bp PSCaVα1 and 325 bp PSEF-1α in tobacco used in plant feeding bioassays. The results revealed significant insect mortality 96% PSCaV-α1 and 46% PSEF-1α and several allied phenotypic deformities like stiffing, melanization and shedding of cuticle, arrested growth and reduced fecundity in P. solenopsis Tinsley were consonant with declined expression of targeted genes in insects feeding corresponding dsRNAs duly elaborated by a semi-quantitative RT-PCR. The specificity of dsRNAs 152 bp PSCaV-α1 and 325 bp PSEF-1α dsRNAs derived from P. solenopsis Tinsley and 402 bp Arginine kinase (AK) from A. gossypii Glover was checked through feeding bioassays in non-target insect species like M. persicae Sulzer, H. virescens and S. littoralis. The results revealed non-significant effect of 152 bp PSCaV-α1 and 325 bp PSEF-1α and significant effect of AK in adult aphid in terms of mortality, nymphs produced and survived, larval mortality and dry weight gain in tested insect species. The cumulative results of this study highlighted the significance of PVX vector as valuable genetic tool for preliminary screening and evaluation of candidate insecticidal proteins and RNAi gene constructs. This study will enhance the efforts of biologists in a way to screen the candidate insect genes and design the complementary transgenic crop protection strategies.
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لومڑی دی چالاکی تے کاں دی بے وقوفی

لومڑی دی چالاکی تے کاں دی بے وقوفی

اک دفعہ دا ذکر اے کہ اک کاں نوں کسے حلوائی دی دکان توں اک پنیر دا تکڑا ملیا۔ کاں نے اوس نوں اپنی چنج نال گھٹ کے پھڑیا تے اڈ کے درخت دی اُچی تھاں اُتے بہہ گیا۔ دل ای دل وچ بہت خوش ہویا کہ اج کھان نوں بڑے دناں بعد چنگی شے ملی اے۔ اوہ خوشی وچ کھاون دا سوچ ای رہیا سی کہ اوہدر اک لومڑی آ گئی۔ کاں کول پنیر دا ٹوٹا ویکھ کے لومڑی دے منہ وچ پانی آ گیا۔ اوہ سوچن لگی کہ کویں ایہہ ٹوٹا کاں کولوں کھو لیا جاوے۔ لومڑی اپنی چالاکی پاروں سارے جانوراں وچ مشہور اے۔ اوس دے دماغ وچ اک تجویز آئی۔ اوہ کاں دی خوشامد کرن لگی۔ اوس آکھیا۔

’’بھرا کاں! توں اک بہت سوہنا پرندہ ایں۔ تیرے پر کنے سوہنے نیں تے تیرا رنگ کناں چمکیلا اے۔۔۔ میرے خیال وچ تیری آواز وی رنگ تے پراں وانگ بہت سوہنی ہووے گی۔ تیری بڑی مہربانی ہووے گی کہ توں مینوں اک سریلا گیت سنا دیویں۔‘‘

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وسائل تشكيل الصورة للموت والحياة عند جبران خليل جبران

The concept of imagery plays an important role in literacy, critical and rhetorical studies. Different research studies describe the reality of imagery its role and methods of its formulation in literary work. Imageries are among those literary tools, through which writers share their experiences. The writer uses different literary tools to formulate imagery and to share their feelings with readers. These tools are simile, metaphor, sensuousness, personification and oxymoron etc. Khalil Jubran was among those writers who frequently used imagery in his work to reflect his inner feelings in a better way. This research study was focused only on the use of imagery tools in his work for the formulation of life and death.

Status of Antioxidant Defenses in Response to Exercise & Antioxidant Therapy in Knee Osteoarthritic Patients.

The aging population and increasing cost of health care will greatly impact on future health care system and the methods used to treat conditions like osteoarthritis. With numerous treatment options for osteoarthritis, decisions about treatment should ultimately be made between the patient and the physician and consider factors such as the degree of pain and incapacitation, potential drug interactions, as well as medical reimbursement. Knee osteoarthritis is a difficult condition to treat. The recent, American college of rheumatology guidelines highlights the major pharmacologic and non-pharmacologic treatments available; other international guidelines supplement these recommendations. There is good evidence for the use of medications such as acetaminophen, topical and oral nonsteroidal anti-inflammatory drugs, and tramadol; injectables including viscosupplementation and intra-articular steroids; lifestyle interventions such as education, exercise, weight loss, and mineral baths; and braces such as a knee brace, knee sleeve, and shoe inserts. There are many controversial treatments needing more study. Ideally, treatment should involve the use of multiple modalities to allow for synergy of interventions. In the future, there is the potential for disease modifying interventions but none have been studied well enough to make recommendations at this time. These disease-modifying interventions would expand the non operative treatment options available for knee osteoarthritis. The present study is unique in the area because as per evidence, this is the first study to prove the efficiency of the exercise and antioxidant supplementation effects alone and when given in combination as an exclusive treatment, in the improvement of the knee osteoarthritis. The effects of exercise, antioxidant supplements, and the combination of both have been observed in osteoarthritic patients .The result attained in this study encouraged us to conclude that in treating osteoarthritis ,collective outcomes of the two, potentiate each other becoming a lot more than their individual effects.It shall have considerable effects on community sociably and economically. Forthcoming idea for molecular biology of superoxide dismutase under the consequences of various treatments will surely open the immense perspective of the research. But further research and evidences are necessary for efficacy of exercises and other treatment additions of antioxidant supplement therapy in osteoarthritis of knee.