Identification and characterization of RAPD–SCAR markers linked to glyphosate-susceptible and -resistant biotypes of Eleusine indica (L.) Gaertn
- 333 Downloads
Eleusine indica is one of the most common weed species found in agricultural land worldwide. Although herbicide-glyphosate provides good control of the weed, its frequent uses has led to abundant reported cases of resistance. Hence, the development of genetic markers for quick detection of glyphosate-resistance in E. indica population is imperative for the control and management of the weed. In this study, a total of 14 specific random amplified polymorphic DNA (RAPD) markers were identified and two of the markers, namely S4R727 and S26R6976 were further sequence characterized. Sequence alignment revealed that marker S4R727 showing a 12-bp nucleotides deletion in resistant biotypes, while marker S26R6976 contained a 167-bp nucleotides insertion in the resistant biotypes. Based on these sequence differences, three pairs of new sequence characterized amplified region (SCAR) primers were developed. The specificity of these primer pairs were further validated with genomic DNA extracted from ten individual plants of one glyphosate-susceptible and five glyphosate-resistant (R2, R4, R6, R8 and R11) populations. The resulting RAPD–SCAR markers provided the basis for assessing genetic diversity between glyphosate-susceptible and -resistant E. indica biotypes, as well for the identification of genetic locus link to glyphosate-resistance event in the species.
KeywordsGoosegrass Molecular marker Insertion–deletion Random amplified polymorphic DNA Sequence characterized amplified region
This project was funded under Fundamental Research Grant Scheme (VOT: 59046) from the Ministry of Higher Education (MOHE) and ScienceFund (Project No: 05-01-12-SF0006) from the Ministry of Agriculture (MOA), Malaysia.
- 3.Heap I (2012) International Survey of Herbicide Rresistance Weeds. http://www.weedscience.org. Accessed 10 May 2012
- 4.McAlister FM, Holtum JAM, Powles SB (1995) Dinitroaniline herbicide resistance in rigid ryegrass (Lolium rigidum). Weed Sci 43:55–62Google Scholar
- 6.Itoh K, Azmi M, Ahmad A (1990) Paraquat resistance in Amaranthus lividus and Conyza sumatrensis in Malaysia. In: Proceedings of the 3rd tropical weed science conference (Kuala Lumpur, 4–6 Dec 1990). Malaysian Plant Protection Society, Kuala Lumpur, Malaysia, pp 489–493Google Scholar
- 7.Marshall G, Kirkwood RC, Leach LE (1993) Comparative studies on graminicide-resistant and susceptible biotypes of Eleusine indica. Weed Res 36:177–185Google Scholar
- 12.Powles SB, Lorraine-Colwill DF, Dellow JJ, Preston C (1998) Evolved resistance to glyphosate in rigid ryegrass (Lolium rigidum) in Australia. Weed Sci 46:604–607Google Scholar
- 17.Gaines TA, Zhang W, Wang D, Bukin B, Chisholm ST, Shaner DL, Nissen SJ, Patzoldt WL, Tranel PJ, Culpeper AS, Grey TL, Webster TM, Vencill WK, Sammons RD, Jiang J, Preston C, Leach JE, Westra P (2010) Gene amplification confers glyphosate resistance in Amaranthus palmeri. Proc Natl Acad Sci 107:1029–1034CrossRefPubMedGoogle Scholar
- 25.Masni Afiza M, Nik Marzuki S, Salmijah S, Ismail BS (2008) Studies on the differentially expressed gene in goosegrase (Eleusine indica [L.] Gaertn) resistant to glyphosate using reverse transcriptase-polymerase chain reaction (RT-PCR) approach. Adv Nat Appl Sci 2:1–5Google Scholar
- 36.Strebig JC, Rudermo M, Jensen JE (1993) Dose-response curves and statisitical models. In: Streibig JC, Kudsk P (eds) Herbicide bioassay. CRC Press, Boca Raton, pp 29–56Google Scholar
- 37.Zimdahl RL (2007) Fundamentals of Weeds Science. Elsevier Academic Press, San DiegoGoogle Scholar