Serological, Molecular Characterization and Diagnostic Methods of Groundnut Bud Necrosis Virus Infecting Onion (Allium cepa L.) in South India

Asadhi, Sujitha; Veera, Bhaskara Reddy Bommu; Yeturu, Sivaprasad; Rayalcheru, Usha

doi:10.3923/ajppaj.2016.29.35

Cited by 1 publication

(1 citation statement)

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“…For the cDNA synthesis of CMV, 20 µl reaction mixture (9 µl- sterile water; 4 µl- 5× reaction buffer; 2 µl- dNTPs; 1 µl- random primer; 1 µl- reverse transcriptase; 1 µl- RNase inhibitor; 1 µg-total RNA) was used (RevertAid, Fermentas, India) and incubated at 42 °C for 60 min followed by 70 °C for 5 min 46 . For tospovirus, the reaction mixture contains total RNA, random primer and nuclease free water up to 12.5 µl were incubated at 65 °C for 5 min followed by 5× reaction buffer, 10 mM dNTP mix, RNase inhibitor and reverse transcriptase were added and incubated for 1 h at 42 °C 47 . The presence of CMV was confirmed through RT-PCR with CMV coat protein gene specific primer (RsCMV-F and RsCMV-R) with a cyclic condition of 94 °C for 2 min followed by 94 °C for 30 s of denaturation, 59 °C for 30 s of annealing, 72 °C for 60 s with final extension of 72 °C for 10 min 48 .…”

Section: Methodsmentioning

confidence: 99%

Co-existence of chlorosis inducing strain of Cucumber mosaic virus with tospoviruses on hot pepper (Capsicum annuum) in India

Vinodhini

Rajendran

Abirami

et al. 2021

Sci Rep

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Cucumo- and tospoviruses are the most destructive viruses infecting hot pepper (chilli). A diagnostic survey was conducted to assess the prevalence of cucumo and tospoviruses in chilli growing tracts of Tamil Nadu. Infected plants showing mosaic with chlorotic and necrotic rings, veinal necrosis, mosaic mottling, leaf filiformity and malformation were collected. Molecular indexing carried out through reverse transcription polymerase chain reaction (RT-PCR) with coat protein gene specific primer of Cucumber mosaic virus (CMV) and tospovirus degenerate primer corresponding to the L segment (RdRp). Ostensibly, amplifications were observed for both CMV and tospoviruses as sole as well for mixed infections. The sequence analysis indicated that the Capsicum chlorosis virus (CaCV) and Groundnut bud necrosis virus (GBNV) to be involved with CMV in causing combined infections. The co-infection of CMV with CaCV was detected in 10.41% of the symptomatic plant samples and combined infection of CMV with GBNV was recorded in around 6.25% of the symptomatic plants surveyed. The amino acid substitution of Ser129 over conserved Pro129 in coat protein of CMV implies that CMV strain involved in mixed infection as chlorosis inducing strain. Further, the electron microscopy of symptomatic plant samples explicated the presence of isometric particles of CMV and quasi spherical particles of tospoviruses. This is the first molecular evidence for the natural co-existence of chlorosis inducing CMV strain with CaCV and GBNV on hot pepper in India.

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Section: Methodsmentioning

confidence: 99%