Identification of markers tightly linked to tomato yellow leaf curl disease and root-knot nematode resistance by multiplex PCR

Abstract

ABSTRACT. Seven different commercial F 1 hybrids and two F 2 populations were evaluated by multiplex PCR to identify plants that are homozygous or heterozygous for Ty-1 and Mi, which confer resistance to tomato yellow leaf curl disease and root-knot nematode, respectively. The Ty-1 and Mi markers were amplified by PCR and identified by digestion of the amplicons with the TaqI enzyme. The hybrids E13 and 288 were found to be Ty/ty heterozygous plants with 398-, 303-, and 95-bp bands, and B08, 314, 198, and A10 were found to be ty/ty homozygous plants with a 398-bp band; whereas 098 did not give any PCR products. The hybrids E13 and 198 were found to be Mi/Mi homozygous plants with 570-and 180-bp bands, and 288 and A10 were found to be Mi/mi heterozygous plants, with 750-, 570-and 180-bp bands, and B08, 109 and 314 were found to be mi/mi homozygous plants with only a 750-bp band. We additionally developed a multiplex PCR technique for JB-1 and Mi, which confer resistance to tomato yellow leaf curl disease and root-knot nematode. The JB-1 marker identified the genotype of the Ty gene, and the plants ©FUNPEC-RP www.funpecrp.com.br Genetics and Molecular Research 11 (3): 2917-2928 (2012 S.X. Chen et al. 2918 that produced the 400-bp band were ty/ty homozygous plants, whereas the plants that produced 400-and 500-bp bands were resistant to tomato yellow leaf curl disease. We conclude that multiplex PCRs can be used to reproducibly and efficiently detect these resistance genes.

Markers for Rapid Evaluation of Virus Resistance for TYLCV in Tomato, ZYMV and PRSV-W in Zucchini and LMV in Lettuce and Hybrid Seeds in Pumpkin

“…Furthermore, the 93% similarity of the primer amplicon sequence developed with S. lycopersicum chromosome 06 strengthened its closeness to the Ty-1 resistance gene located on the same chromosome. A multiplex PCR using CAPS primers for the JB-1 and Mi genes identified at the same time the genotypes of resistance to the root-knot nematode and TYLCV in the F1 and F2 generations [28]. So, the development of a marker derived from JB-1, which does not require enzymatic digestion, will make easier the screening of TYLCV resistance and incorporation of this allele by marker-assisted selection.…”

Multiplex PCR for Detection of Tomato Yellow Leaf Curl Disease and Root-Knot Nematode Resistance Genes in Tomato (Solanum lycopersicum L.)

“…R and S indicate the position of the bands for the resistance and susceptibility alleles S. chilense, S. habrochaites and S. peruvianum and thus facilitates marker-assisted resistance gene pyramiding. Previously, multiplex PCR tests for tomato resistance gene alleles have been described, but these tests either targeted a smaller number of loci (Chen et al, 2012;Fu et al, 2013) or a different locus combination (Liu et al, 2013). This method showed that the susceptibility alleles at two Ty resistance gene loci in S. chilense accessions were different than those in S. lycopersicum.…”

“…Masuelli et al (2000) described the development of a PCR assay simultaneously detecting the root-knot nematode resistance gene Mi-1 and the Tomato spotted wilt virus resistance gene Sw-5 in tomato. Several other multiplex assays specific for two to three tomato resistance gene alleles have been described (Chen et al, 2012;Fu et al, 2013). Liu et al (2013) disclosed a multiplex PCR assay for Ty-1, Ty-2, Mi-1 and Cf-5 resistance genes, but not for Ty-3 and Ty-5.…”

Identification of microsatellite markers in coffee associated with resistance to Meloidogyne exigua

“…Fuganti et al (2004) identified two microsatellite markers, named Satt 144 and SOYHSP 176, which were correlated with the gall number and infestation scores of M. javanica in soybean. Ruan et al (2009) identified inter-simple sequence repeat markers associated with disease resistance in Hippophae L. Chen et al (2012) used multiplex PCR to identify molecular markers linked to root-knot nematode resistance in tomato, and identified a specific marker (Mi).…”