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Home > Genetic Diversity of Begomoviruses Affecting Diverse Host Plants in Periurban Areas of Lahore

Genetic Diversity of Begomoviruses Affecting Diverse Host Plants in Periurban Areas of Lahore

Thesis Info

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Author

Qurashi, Fasiha

Program

PhD

Institute

University of the Punjab

City

Lahore

Province

Punjab

Country

Pakistan

Thesis Completing Year

2019

Thesis Completion Status

Completed

Subject

Plant Pathology

Language

English

Link

http://prr.hec.gov.pk/jspui/bitstream/123456789/14472/1/Fasiha%20Qurashi%20plant%20patholgy%202019%20uop%20lhr%20prr.pdf

Added

2021-02-17 19:49:13

Modified

2024-03-24 20:25:49

ARI ID

1676726294280

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Plant foliage exhibiting symptoms indicative of begomovirus infection, veinthickening, leaf curling, yellowing and chlorosis, dwarfing and mosaic were collected from around two km away of Peri-urban areas of Lahore from Multan road, University of the Punjab, Quaid-e-Azam campus, Shikhupura, Ferozwala, Wahga border, Sharaqpur and Ferozpur roads in Pakisatan during 2013-2016. Total plant genomic DNA was isolated from leaf tissues by Cetyl Trimethyl Ammonium Bromide (CTAB) method described by Doyle and Doyle, (1990). Universal primers were used to identify the presence of begomovirus and associated DNA-satellite complex (betasatellite, geminivirus associated alphasatellite) and were subjected to rolling circular amplification (RCA). The expected size of PCR products were cloned and sequenced. Specific abutting primers were designed from the available sequences to amplify the full-length begomoviruses. These full-length PCR entities were cloned and sequenced in their entirety. The isolates of Chickpea chlorotic dwarf virus (CpCDV) and Mesta yellow vein virus (MeYVV) with non-cognate Cotton leaf curl Multan betasatellite (CLCuMuB) were used to produce partial repeat constructs for agro-inoculation. All reported samples were Old World (OW) monopartite begomoviruses, showing recombination and were associated with DNA-satellite complex. In this study the OW monopartite begomovirus Cherry tomato leaf curl virus (CToLCV) was associated with Papaya leaf curl betasatellite (PaLCuB) and Tobacco curly shoot alphasatellite, first time isolated from Parthenium hysterophorus in Pakistan (Qurashi et al., 2017). Similarly ornamental infecting begomovirus associated with DNA- satellite complex also reported from Malva parviflora, a new strain of Hollyhock leaf curl virus (HoLCV-Mal) associated with Kenaf leaf curl betasatellite (KLCuB) and two geminivirus associated alphasatellites species; Ageratum enation alphasatellite (AEA) and Ageratum yellow vein India alphasatellite (AYVIA) in the sub family Geminialphasatellitinae and the genus Colecusatellite (Briddion et al., 2018) identified from Malva parviflora. According to old alphasatellite classification (Mubin et al., 2009) Ageratum enation alphasatellite specie was known as Ageratum conyzoides alphasatellite (Sattar et al., 2017). Another weed infecting begomovirus was associated with DNA-satellite complex also reported from woody plant mulbery isolated as Ageratum enation virus (AEV) associated with Papaya leaf curl betasatellite (PLCuB) and associated geminivirus alphasatellite; Guar leaf curl alphasatellite, but according to new alphasatellite classification this geminivirus associated alphasatellite specie is known as Ageratum enation alphasatellite (AEA) in the genus Colecusatellite described in detailed in this study. All these begomoviruses associated with DNA-satellite complex are reported for the first time in Pakistan. In this study there was prevalence of Pedilanthus leaf curl virus (PeLCV) infecting diverse host plants; ornamental Cestrum nocturnum and reported for first time from vegetables Trigonella foenum and Piper nigrum woody plant Albizia lebbeck in Pakistan. Pedilanthus leaf curl virus (PeLCV) with geminivirus associated alphasatellites; Ageratum enation alphasatellite also reported from Trigonella foenum Albizia lebbeck, respectively. Infectivity assay of the partial repeat constructs of mastrevirus; Chickpea chlorotic dwarf virus (CpCDV) and begomovirus; Mesta yellow vein virus (MeYVV) with non-cognate Cotton leaf curl Multan betasatellite (CLCuMuB) were checked. These were infectious to experimental host plant Nicotiana benthamiana, the Koch’s postulates for CpCDV alone, MeYVV alone and with non-cognate CLCuMuB showed severe symptoms. All the experimental results were satisfied and confirmed with PCR, real-time PCR and Southern blot hybridization. Furthermore, infectivity assay of recombinant PeLCV alone was also checked through gen gun method but PeLCV did not produce begomoviral symptoms in Nicotiana benthamiana, PeLCV need associated betasatellite for begomoviral infection.
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