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Home > Use of a Plant Virus Vector for Screening of Gene Constructs With Potential Insecticidal Activity Musarrat Shaheen 2017

Use of a Plant Virus Vector for Screening of Gene Constructs With Potential Insecticidal Activity Musarrat Shaheen 2017

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

Natural Sciences

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9447/1/Musarrat%20Thesis%2014-7-2017.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676727643109

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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. xviii 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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مولانا حیدر حسن خاں محدث ٹونکی

مولانا حیدر حسن صاحب محدث ٹونکی کی وفات
مولانا حیدر حسن خاں صاحب محدث ٹونکی جو تقریباً دس پندرہ برس تک دارالعلوم ندوۃ العلماء لکھنؤ میں شیخ الحدیث رہ کردوسال ہوئے کہ ریاست کی خواہش پر اپنے وطن چلے گئے تھے، افسوس ہے کہ چند روز ہوئے کہ اپنے وطن ہی میں وفات پائی ، محدث مرحوم اورا ن کے بڑے بھائی مولانا محمود حسن خاں صاحب مصنف معجم المصنفین اس وقت کے علماء میں ایسے دو نامور فرد تھے کہ جن کے وجود پر علم و فضل اور ورع و تقویٰ کو ناز تھا، الحمدﷲ کہ ابھی مولانا محمود حسن خاں صاحب ہم میں موجود ہیں، مگر افسوس ہے کہ ان کے چھوٹے بھائی مولانا حیدر حسن خاں صاحب نے اس عالم فانی کو الواداع کہا، ایسے زمانہ میں جب نام کے مولویوں کی تعداد گو کثرت حاصل کررہی ہے، مگر کام کے علماء روز بروز کم سے کم ہوتے جارہے ہیں، مرحوم کی وفات مشرقی علم و فضل کی کائنات میں حادثہ عظیم سمجھی جائے گی۔
مرحوم بڑے جامع العلوم تھے، علوم عقلیہ و نقلیہ و ریاضیہ کے وہ یکساں ماہر تھے، زیادہ تر اپنے بڑے بھائی مولانا محمود حسن خاں صاحب سے پڑھا تھا، حدیث کی سند شیخ حسین صاحب عرب یمنی خزرجی سے حاصل کی تھی، استفادہ باطنی میں بھی ان کا مرتبہ بلند تھا، انہوں نے مکہ معظمہ جاکر حضرت حاجی شاہ امداد اﷲ صاحب رحمتہ اﷲ علیہ سے فیض حاصل کیا تھا، اور ماشاء اﷲ زہدو عبادت بے تکلفی اور تواضع میں بزرگوں کا نمونہ تھے، علم عقلیہ و ریاضیہ میں بھی ان کا درجہ بلند تھا، اور علوم نقلیہ میں وہ ماہر کامل تھے، علم حدیث کو بطرز حنفیہ بہت خوبی سے پڑھاتے تھے، رجال پر ان کی نظر وسیع تھی، ان کے درس کا طریقہ یہ تھا کہ حدیث...

The Lung Mass and Nodule: A Case Series The Lung Mass and Nodule

Lung mass is an abnormal region of 3 cm or more in size present in the lungs mainly due to underlying pulmonary caner. It is usually round, opaque and poorly differentiated on X-ray. Common etiological key players are smoking, exposure to asbestos, radon, however, familial history may also play a role. We presented retrospectively7 cases of lung mass and nodule encountered during our clinical practice. We have discussed their clinical presentation, manifestation, medical history, radiological findings and differential diagnosis. In this case series, most of the patients were young, only 2 cases were older patients. There was one infant one month old, one female child 12 years old, one female 25 years, 2 males, 22 and 21 years, one male of 50 years and another male of 60 years age. Correct diagnosis on the basis of clinical profile, radiological findings and histology may help in proper management and hence, timely treatment of the patient

Studies on the Use of Seeds of Some Indigenous Plants for the Control of Root-Knot Nematodes.

Meloidogyne javanica (Trueb) Chitwood Root knot nematodes invade the roots of plants and cause significant harm to crops. Fifty plant seeds belonging to different families were collected and evaluated for their nematicidal activity. Seeds were screened for nematicidal activity in vitro and in vivo. Aqueous seeds extract of Acacia concinna, Amaranthus paniculatus, Anethum graveolens, Cassia fistula, Cassia holosericea, Leucaena leucocephala, livistona chinensis, Moringa oleifera, Nigella sativa, Sesbania bispinosa and Canna indica showed hundred percent decrease in egg hatching of Meloidogyne javanica. Aqueous seeds extract of Acacia stenophylla, Asteracantha longifolia, Brassica juncea, Caesalpinia pulcherrima, Clitoria ternatea, Conocarpus lancifolius, Cuminum cyminum, Foeniculum vulgare, Papaver somniferum, Peltophorum pterocarpum, Sesbania bispinosa, Solanum miniatum and Trigonella foenum-graecum showed 100 percent juvenile mortality in vitro. Methanolic seeds extract of Leucaena leucocephala, Cassia fistula, Peltophorum pterocarpum, Canna indica, Brassica juncea, Cassia holosericea, Acacia concinna, Caesalpinia pulcherrima, Linum usitatissimum and Thespesia populnea also showed significant reduction in egg hatching and larval mortality of Meloidogyne javanica in vitro In green house experiment against Meloidogyne javanica aqueous extracts of seeds powder were found to be effective either by drenching or seed treatment were Sesbania bispinosa, Solanum nigrum, Clitoria ternatea, Brassica juncea, Acacia stenophylla, Foeniculum vulgare, , Canna indica, Cuminum cyminum, Ricinus communis, Trigonella foenum-graecum, Pithecellobium dulce, Linum usitatissimum, Moringa oleifera, Anethum graveolens, Asteracantha longifolia, Trachyspermum ammi, Canna indica, Livistona chinensis, Clitoria ternatea, Pithecellobium dulce Thespesia populnea and Coriandrum sativum. Organic amendment of soil was done by mixing seed powder in soil @ 1 % w/w after 3 weeks of organic amendment seeds were sown. Amendment with seeds powder of Asteracantha longifolia, Amaranthus paniculatus, Caesalpinia gilliesii, Ricinus communis, Sesbania bispinosa, Parkinsonia aculeata, Tamarindus indica, Eruca sativa, Acacia stenophylla, Albizia lebbeck, Nigella sativa, Piper nigrum, Ipomoea pes-caprae, Brassica juncea, Prunus avium, Solanum nigrum, Foeniculum vulgare, Thespesia populnea, Sapindus trifoliatus, Cordia sebestena, Moringa oleifera, Tribulus terrestris, Clitoria ternatea, Cassia holosericea, Senna alata, Peltophorum pterocarpum, Delonix regia, Coriandrum sativum, Linum usitatissimum, Adenanthera pavonina, Leucaena leucocephala, pithecellobium dulce, Roystonea regia, Datura innoxia, Conocarpus lancifolius, Anethum graveolens, Trigonella foenum-graecum, Trachyspermum ammi, Eragrostis abyssinica, Cuminum cyminum, Sinapis alba, Caesalpinia pulcherrima, Pongamia pinnata, Cassia fistula Acacia concinna and Acacia nilotia appeared to be potent against root knot nematode and increased the growth of okra and cowpea plants. When organic amendment of soil with Acacia concinna, Canna indica, Foeniculum vulgare, Sesbania bispinosa and Solanum nigrum seeds meals at different concentrations of 0.3, 0.5 and 1% w/w of soil was performed. Most significant results were obtained in the treatments with Sesbania bispinosa and Solanum miniatum at organic amendment of 1 percent w/w in soil in both crops. This concentration was selected for further studies and field experiment. Combined use of Sesbania bispinosa and Solanum nigrum seed meals with chemical and biological antagonist also provided significant results as there enhancement in growth parameters and significant decrease in infection of Meloidogyne javanica and knot count in roots. Most promising results appeared when seed meals were mixed with Beauveria bassiana and Furadan. Preliminary phytochemical screening from ethanolic extract of seeds with strong nematicidal activity was performed. Phytochemical tests revealed presence many secondary metabolites flavanoid, terpenoid, steroid, tannin, carbohydrate, phenol, alkaloid, saponin, coumarin, glycolside and phlobatannin in seeds. Organic amendment of soil in field condition with Solanum nigrum and Sesbania bispinsosa showed significant decrease in the infection of root knot nematodes and increase in okra and cowpea plants growth.