Utility of MtCOI polymerase chain reaction‐restriction fragment length polymorphism in differentiating between Q and B whitefly Bemisia tabaci biotypes

Abstract

The invasive, insecticide-resistant, Q whitefly biotype, has gradually spread to other countries including the US via human-mediated movement of plant materials. We assessed the utility of the VspI-based mtCOI (mitochondrion cytochrome oxidase I) polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique as a rapid, cost-effective, and reliable alternative for differentiating the Q from the dominant B biotype in Arizona. Using the standard mtCOI gene sequencing and mtCOI PCR-RFLP techniques, we biotyped eight whitefly strains of five individuals each collected from poinsettia and cotton at different locations in Arizona. Complete concordance was observed between the two methods, with three strains being identified as the Q biotype and five samples as the B biotype. We also scanned the mtCOI gene sequences for VspI polymorphisms in the B and Q biotype whiteflies currently available in the GenBank database. This global screening revealed the existence of three and four VspI polymorphic types for the Q and B biotypes, respectively. Nevertheless, all three VspI polymorphic Q biotype whiteflies shared a common and unique VspI site that can be used to differentiate Q biotype from the four VspI polymorphic B biotype whiteflies identified. These results demonstrate that the VspI-based mtCOI gene PCR-RFLP provides a reliable diagnostic tool for differentiating the Q and B biotype whiteflies in the US and elsewhere.

Use of Mitochondrial Cytochrome Oxidase I Polymerase Chain Reaction-Restriction Fragment Length Polymorphism for Identifying Subclades of Bemisia tabaci Mediterranean Group

“…PCR ampliÞcations began with 94ЊC denaturation for 3 min, followed by 35 cycles of 94ЊC denaturation for 1 min, 52ЊC annealing for 1 min, and 72ЊC extension for 2 min, and a Þnal 72ЊC extension for 10 min. Half of each PCR products were used to differentiate MED from MEAM1 by using VspI-based mtCOI PCR-RFLP (Ma et al 2009). When a sample was identiÞed as a MED individual by VspI-based mtCOI PCR-RFLP (Ma et al 2009), the remaining half of its PCR products were then digested at 37ЊC for 2 h with 2 U of AluI, a restriction endonuclease that cleaves DNA at AGCT sites.…”

“…In total, 120 mtCOI sequences (545 bp) were generated by sequencing the mtCOI PCR products of the MED samples detected in Arizona during 2004Arizona during Ð2008 (27 in 2004Arizona during , 30 in 2006Arizona during , 27 in 2007Arizona during , and 35 in 2008 as described in Ma et al (2009). In addition to the 120 MED mtCOI sequences, 1,839 worldwide B. tabaci mtCOI sequences in total, including from the United States, were retrieved from GenBank (http://www.ncbi.nlm.nih.gov/) on 21 March 2011.…”

“…There are several diagnostic techniques available for differentiating different B. tabaci biotypes (Khasdan et al 2005, Bosco et al 2006, Ko et al 2007). We have recently conÞrmed that the VspI-based mtCOI polymerase chain reaction (PCR)-restriction fragment length polymorphism (RFLP) developed by Khasdan et al (2005) and Horowitz et al (2005) can reliably differentiate MEAM1 and MED whiteßies in the United States (Ma et al 2009). Here, we report the development of further diagnostic methods to distinguish the Q1 and Q2 for each other and MEAM1.…”

Study on species composition of Bemisia tabaci (Gennadius, 1889) (Hemiptera: Aleyrodidae) on cotton in Çukurova plain, Turkey

“…PCR-RFLP was reported as a simple, quick, cost-effective and reliable alternative method to distinguish B. tabaci species (Khasdan et al 2005;Ma et al 2009;Chu et al 2012). This method was applied to support results of sequence analysis and differentiating the MED and MEAM1 species of B. tabaci populations.…”

A critical evaluation of the effects of plant essential oil formulations against silverleaf whitefly, Bemisia tabaci, B biotype (Gennadius) and its natural enemies

“…The main genetic techniques were allozymes (Gunning et al 1997), RAPD PCR (randomly amplified polymorphic DNA polymerase chain reaction) (De Barro and Driver 1997), AFLP (amplified fragment length polymorphism) (Cervera et al 2000), rITS1 (ribosomal integenic transcribed spacer 1) (De Barro et al 2000) and mitochondrial DNA markers, mt16s and mtCo1) ( Frohlich et al 1999;Ma et al 2009). From the above studies of the phylogenetic characteristics of B. tabaci biotypes, De Barro et al (2011) have listed 36 identified biotypes worldwide.…”