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Genomic Variability Among Barley Yellow Dwarf Virus-Pav Infecting Wheat Crop in Pakistan

Thesis Info

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Author

Qadir, Abdul.

Program

PhD

Institute

The University of Agriculture

City

Peshawar

Province

KPK

Country

Pakistan

Thesis Completing Year

2018

Thesis Completion Status

Completed

Subject

Genetics

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/9292/1/Abdul%20Qadir_Plant%20%26%20Env.%20Protection_2018_UAP_PRR.doc

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726331457

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Wheat (Triticum aestivum L.) is the major staple grain food of Pakistan and is prone to many fungal, bacterial and viral diseases. Diseases caused by vriuses are among the biotic factors inflicting huge economic losses. Every year Barley yellow dwarf virus (BYDV) causes substantial losses to wheat crop. A total of 210 samples showing typical barley yellow dwarf virus symptoms were collected from different wheat growing area of Pakistan. The DAS-ELISA technique was used to identify the existing serotype of BYDV. The BYDV-PAV was serologically identified as the most prevalent strain in Pakistan. The overall infection rate in symptomatic plants of BYDV-PAV was 43% whereas highest infection was recorded in Muzaffar Garh area of Punjab province. Different plants species viz. oat (Avena sativa), maize (Zea mays), johnson grass (Sorghum halepense) and Italian rye (Lolium multiform) in the cropping system was tested for BYDV-PAV infection. The maize and grasses showed the highest infection rate of 70% while oats showed comparatively less infection percentage (63%). Total RNA was extracted by Tri- reagent from BYDV-PAV ELISA positive samples. Coat protein gene of BYDV-PAV was amplified by reverse transcriptase polymerase chain reaction (RT-PCR) using specific primers set and a product of approximately 600 bp was obtained. The overlapping strategy was employed for which 12 primers set were designed, for complete genome sequencing. A maximum of 5654bp complete genome size of Pakistani isolate of BYDV-PAV was obtained after joining contigs. The complete genome sequences were compared with other world isolates of BYDV-PAV such as Asia, Europe and USA by using basic local alignment search tool (BLAST) available on National Center for Biotechnology Information (NCBI) data base. The phylogenetic analysis was done by using Molecular Evolutionary Genetics Analysis (MEGA) Program. The gene wise inter isolate comparison among four Pakistani isolates of wheat was made. Between the RdRp/POL genes (i.e. P1 and P2), P2 gene showed higher identity (94.7-99.8%) as compared with P1 gene. The P3 coat protein gene was found the most conserved region with maximum of 99.2-100% among the isolates. The nucleotide sequence of P4 (putative movement protein gene) has showed identity of 98.6-100% among inter isolate of BYDV-PAV. The nucleotide sequence of aphid transmission gene/RTD (P5) has shown identity of 91.1-99.9% among inter isolate. Similarly the P6 gene, whose function is still unknown, has shown sequence identity of 89.4-99.9% among inter isolate. Similarity index of un-translated region (UTR) i.e. 5'', 3'' and intergenic regions were 91.4-100%, 93.9-100%, 97.6-100% (P2-P3) respectively while for P5-P6 intergenic region was 93.8-100%. The overall complete genomic comparison among four Pakistani Isolates (KT 252975, KT 252976, KT 252977and KT 252978) has shown divergence ranges from 0.14% to 6.9%. However these isolates have shown identity of 74.4 to 99.9% with other BYDV-PAV isolates of the world. It is concluded from this study that Pakistani isolates of BYDV-PAV are more closely related to USA, Europe and Japan but are distinct from Chinese isolates. Moreover the German isolate seems establishing an interlink bridge between PAV-I and PAV- II clusters based on phylogenetic tree analysis. The sequences of Pakistani isolates clearly indicated their identity as BYDV-PAV.
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محمد علی جناح [قائد اعظم]

محمد علی جناح
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سپردم بتومایۂ خویش را
/تودانی حسابِ کم وبیش را
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کھیل ہیں دورِآسمانی کے
چنانچہ مرحوم کہابھی کرتے تھے کہ ’’ہندومجھ کواپنا دشمن سمجھتے ہیں لیکن اگر پاکستان بن گیا تووہ ہمیشہ میرے احسان مند رہیں گے اورمیرے مرنے کے بعد ان کومحسوس ہوگا میں ان کادشمن نہیں سچا دوست تھا۔‘‘
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فروغ تعلیم: اسوہ رسول کی روشنی میں

Education has been considered the core value of human life. Religion and education are interrelated. The preaching of religion depends upon the education and training. Our holy Prophet (SAW) was an ideal educationist not only of his times but for the future generations as well. The article deals with the importance of education in the light of prophetic model. Prophet Mohammad (SAW) has emphasized on education and guidance of Muslims. He has established many educational policies for the Muslims which include Treaty of Madina, brotherhood of Muslims, construction of masajid, establishment of Suffa. These initiatives provided a ground work for future generations in shaping their educational syllabus and policies.

Substrate Dependent Microbial Biosynthesis of Auxins and Their Effect on Growth and Yield of Mung Bean Vigna Radiata L.

Rhizobia are capable of promoting the growth of legumes through a number of mechanisms of action including phytohormone production. Auxin being a plant hormone is produced by rhizobia and this ability increases many folds in supplementation with some suitable physiological precursor like L-tryptophan. Substrate-derived auxins have the ability to promote plant growth by supplementing sub-optimal auxin levels in legume plants like mung bean. So, this project was designed to evaluate the effectiveness of rhizobia under axenic conditions in supplementation with L-tryptophan on the growth and yield of mung bean crop. To test this hypothesis, a series of laboratory, wire house and field experiments were conducted. Rhizobium phaseoli were isolated from different locations of Faisalabad from nodules of mung bean crop. Forty four rhizobial cultures were isolated from mung bean nodules and tested for auxins biosynthesis in vitro colorimetrically and confirmed by high performance liquid chromatography (HPLC). Rhizobial isolates varied in auxin biosynthesis ranging from 8.8 μg IAA equivalents mL -1 to 38.3 μg IAA equivalents mL -1 without L-tryptophan. Auxin biosynthesis by rhizobial isolates in supplementation with L-tyrptophan, increased many folds (1.1 to 7.9 folds) and N42 produced maximum auxins in L-tryptophan supplemented media. Different environmental factors like substrate (L-Trp) and C-source (glucose), pH, incubation temperature, incubation time and aeration (static vs. shaking) significantly affected auxin production. Growth pouch experiments were conducted in the laboratory using 34 rhizobial isolates for improving growth of mung bean seedlings using three mung bean cultivars i.e. NM-92, NM-98 and NM-2006 under axenic conditions. Fifteen isolates were selected through screening in jars using principal component scoring method. Three most efficient rhizobial isolates i. e. A23, N12 and N42 were tested alone and in combination with different L-TRP concentrations (10 -3 , 10 -4 and 10 -5 M) to test the hypothesis of substrate dependent auxin production for improving the growth of mung bean (var: NM-92, NM-98 and NM-2006) under axenic conditions. These isolates were further tested alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp for improving growth and nodulation of mung bean. Finally these three rhizobial isolates i.e. A23, N12 and N42 along with 10 -3 , 10 -4 , 10 -5 M L-Trp concentrations were further used in pot and field experiments using mung bean variety NM-2006. Rhizobial isolates A23, N12 and N42 alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp concentrations were tested in pots at the wirehouse, Institute of Soil and Environmental Sciences, University of Agriculture, Faisalabad. Results of pot experiments revealed that separate application of rhizobia significantly increased growth parameters but the combined application of rhizobia and L-tryptophan further significantly increased yield and nodulation. Field trials were conducted at two sites these rhizobial isolates A23, N12 and N42 alone and in combination with 10 -3 , 10 -4 and 10 -5 M L-Trp concentrations were tested at two N levels i. e. half and full recommended dose of N fertilizer. Results revealed that rhizobial inoculation alone significantly increased growth parameters (fresh biomass, grain yield, shoot length, root length, number of pods plant -1 , and number of grains pod -1 , 1000-grain weight, number of nodules plant -1 , fresh and dry weight of nodules, NPK contents in grain) of mung bean variety NM-2006. But the combined application of rhizobia along with L-Trp (10 -4 and 10 -5 M) further increased the mung bean yield and nodulation. This implies that substrate dependent microbial biosynthesis of auxins as a result of precursor (L-Trp)-inoculum (rhizobia) interactions can be successfully used for improving the growth and yield of legumes.