Use of the random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) to detect DNA polymorphisms in aphids (Homoptera: Aphididae)

1992 ◽  
Vol 82 (2) ◽  
pp. 151-159 ◽  
Author(s):  
William C. Black ◽  
Nancy M. DuTeau ◽  
Gary J. Puterka ◽  
James R. Nechols ◽  
Jennifer M. Pettorini

AbstractWe have used a new technique to identify discrete genetic markers in aphids, a family in which biochemical and morphological genetic polymorphisms are rare. The new technique uses the polymerase chain reaction (PCR) to amplify random regions of aphid genomes (random amplified polymorphic DNA) and has been termed RAPD-PCR. We demonstrate the use of the technique in revealing genetic variation in four aphid species, the greenbug (Schizaphis graminum (Rondani)), the Russian wheat aphid (Diuraphis noxia (Mordvilko)), the pea aphid (Acyrthosiphon pisum (Harris)), and the brown ambrosia aphid (Uroleucon ambrosiae (Thomas)). In contrast with allozyme surveys, RAPD-PCR revealed large amounts of genetic variation among individuals in each of these species. Variation was detected among biotypes, populations, colour morphs and even individuals on a single plant. We also explored the utility of RAPD-PCR in the detection and identification within aphid bodies of two endoparasitic wasps, Diaeretiella rapae (McIntosh) and Lysiphlebus testaceipes (Cresson). The use of RAPD-PCR in species diagnostics, parasitoid detection, and population studies is discussed.

1999 ◽  
Vol 131 (2) ◽  
pp. 229-230 ◽  
Author(s):  
C.K. Chan ◽  
D.J. Petersen ◽  
T.C. Vrain

Extraction of DNA from whole aphids, in combination with random amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) (Williams et al. 1990) markers can detect interspecific and intraspecific genetic variation (Black et al. 1992; Cenis et al. 1993). However, these techniques entail destructive sampling of fresh or preserved specimens. To allow experimental replication from a single sample while preserving the same aphid for morphometrical or karyotyping analyses, we describe a technique for RAPD-PCR using DNA from single aphid embryos. We evaluated the usefulness and reliability of single-embryo analysis, using four species of our laboratory colonies, namely Acyrthosiphon pisum (Harris), Aphis fabae Scopoli, Aphis frangulae group, and Aphis gossypii Glover.


2011 ◽  
Vol 6 (8) ◽  
pp. 35-40
Author(s):  
Sangita Shrestha ◽  
Jaishree Sijapati ◽  
Neesha Rana ◽  
Diwa Malla ◽  
Prabha Regmi ◽  
...  

Of the 30 species (including five varieties) of the genus Swertia in Nepal, nine have been reported to possess medicinal properties. Among these, S. chirayita is the most valuable species, with high demand in domestic and international markets. Nepal’s S. chirayita and related species are being recklessly exploited for commercial purposes. Two problems that have emerged with this lucrative market are (a) adulteration and fraudulent labeling of S. chirayita, and (b) depletion of S. chirayita and allied species from their natural habitats. To address the problem of adulteration and conservation, we studied molecular genetic diversity in S. chirayita populations and developed a molecular diagnostic tool for the purposes of authentication. We studied intra-specific genetic diversity in S. chirayita using Polymerase Chain Reaction (PCR)-based Random Amplified Polymorphic DNA (RAPD) technique. As a preliminary step, we identified optimal RAPD-PCR reaction and cycling conditions by varying PCR reaction parameters such as concentration of template DNA, MgCl2, dNTPs, primer, Taq DNA polymerase and RAPD-PCR programs. The optimized PCR reaction and cycling conditions were then used in subsequent RAPD profiling experiments for the study of genetic diversity within S. chirayita populations from various geographical locations. Genetic diversity characterization of S. chirayita populations at the molecular level would furnish information with significant applications in the conservation and sustainable utilization of S. chirayita and its allied species in Nepal. Key words: Polymerase Chain Reaction, Random Amplified Polymorphic DNA, DNA fingerprinting, genetic diversity DOI: http://dx.doi.org/10.3126/hjs.v6i8.2699 Himalayan Journal of Sciences Vol.6 Issue 8 2010 pp.35-40


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