Paper Title :Manipulation of Different Molecular Technology Including ITS, RELP, Scar Primer and Multiplex Tests for Root Knot Nematode Identification

Author :Sanaa. A. Haroon

Article Citation :Sanaa. A. Haroon , (2018 ) " Manipulation of Different Molecular Technology Including ITS, RELP, Scar Primer and Multiplex Tests for Root Knot Nematode Identification " , International Journal of Advances in Science, Engineering and Technology(IJASEAT) , pp. 34-39, Volume-6,Issue-1,Special Issue - 1

Abstract : Root knot nematode is considered as one of the most important nematode all over the world in Egypt it was found out that theses nematode attack the root system of most vegetables, fruits and Feld crops causing significance damages and economic loss .Using the morphological characterization isn't enough for nematode identification to species. So different molecular technology was used to identify different isolated of these nematode for fast and more accurate results. The internal transcript spacer (ITS) and restriction fragment length polymorphism (RFLP) of Ribosomal DNA (rDNA) sequences were used to distinguish between different major species of root knot nematode. DNA fragments containing the internal transcribed spacer (ITS) rDNA were amplified from DNA genome of different Meloidogyne spp. isolates from different governorate in Egypt. Ten females were used to amplify PCR product. When primers 5368 and 5367 were used for amplification of the ITS region, every isolate from the Meloidogyne spp. gave one major product of approximately 760 bp. When ITS regions of all isolates were digested with restriction enzymes, by using RFLP test (Restriction fragment length polymorphisms), the size of the DNA fragment using Hind III was strong band at 560 bp and another weak band at 200 bp that is typical for M. incognita and M. javanica with Hinf I, bands of 440 and 320 pb were obtained while 520, 240 bp bands were cleared when EcoRl are used. Four bands of 220, 200, 180 and 160 bp were obtained when I restriction enzyme was used. In multiplex test, a fragment of 415 bp was obtained when M. incognita was used as template (positive control), that confirming the fact that most nematodes isolates in Egypt are M. incognita Keywords - Meloidogyne, multiplex, PCR, ribosomal DNA, ITS, RFLP, Scar , restriction enzymes .

Type : Research paper

Published : Volume-6,Issue-1,Special Issue - 1

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