Showing papers by "Mike J. Wilkinson published in 2008"

High-resolution melt analysis to identify and map sequence-tagged site anchor points onto linkage maps: a white lupin (Lupinus albus) map as an exemplar.

Abstract: Summary • The provision of sequence-tagged site (STS) anchor points allows meaningful comparisons between mapping studies but can be a time-consuming process for nonmodel species or orphan crops. • Here, the first use of high-resolution melt analysis (HRM) to generate STS markers for use in linkage mapping is described. This strategy is rapid and low-cost, and circumvents the need for labelled primers or amplicon fractionation. • Using white lupin (Lupinus albus, x = 25) as a case study, HRM analysis was applied to identify 91 polymorphic markers from expressed sequence tag (EST)-derived and genomic libraries. Of these, 77 generated STS anchor points in the first fully resolved linkage map of the species. The map also included 230 amplified fragment length polymorphisms (AFLP) loci, spanned 1916 cM (84.2% coverage) and divided into the expected 25 linkage groups. • Quantitative trait loci (QTL) analyses performed on the population revealed genomic regions associated with several traits, including the agronomically important time to flowering (tf), alkaloid synthesis and stem height (Ph). Use of HRM-STS markers also allowed us to make direct comparisons between our map and that of the related crop, Lupinus angustifolius, based on the conversion of RFLP, microsatellite and single nucleotide polymorphism (SNP) markers into HRM markers.

Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.)

Abstract: Fang, Jong-Yi, Wetten, A., Adu-Gyamfi, R., Wilkinson, M., Rodriguez-Lopez, C. (2009). Use of secondary somatic embryos promotes genetic fidelity in cryopreservation of cocoa (Theobroma cacao L.). Agricultural and Food Science, 18, (2 (Special issue)), 152-159. Sponsorship: COCOA RESEARCH UK IMPF: 00.92 RONO: 00

Methods of producing haploid and doubled haploid oil palms

Abstract: The present invention relates to haploid oil palm plants and homozygous doubled haploid oil palm plants. The invention also relates to methods for producing and selecting haploid and doubled haploid plants. More particularly, but not exclusively, the method may be used for selecting haploid and doubled haploid oil palm plants. Haploid and doubled haploid plants are selected by a large-scale screening based on a combination of the phenotype with the use of molecular methods combined with flow cytometry techniques to identify haploid and doubled haploid plants. More particularly, a method for selecting haploid and doubled haploid plants is described comprising: (a) germinating seeds; (b) selecting seedlings with atypical phenotype; (c) assessing heterozygosity using markers; (d) isolating cells from the seedlings and determining the DNA content of the cells; and (e) isolating and purifying the DNA and using defined molecular markers to characterise the genotype of the plant. The haploid oil palm plants may be used for producing homozygous doubled haploid oil palms: doubled haploids may be intercrossed to produce uniform F1 hybrids of superior properties.

Development and characterisation of microsatellite markers for the fungus Lasiodiplodia theobromae

Abstract: Lasiodiplodia theobromae is an important fungal pathogen of higher plants from tropical and sub-tropical regions. The fungus infects divergent hosts in a wide range of environmental conditions, suggesting that it is highly variable. The aim of this study was to develop new polymorphic microsatellite markers from a Brazilian isolate of L. theobromae that can be used in population studies of this and related fungi. The nine microsatellite markers developed included six that revealed allelic polymorphisms among nine isolates of the disease collected from infected plants in Brazil. Preliminary evaluation of the markers suggested substantial genetic variability among Brazilian L. theobromae populations. These markers have potential utility for evolutionary and epidemiologic studies of this fungus.

High-resolution melt analysis for SNP discovery, linkage mapping and analysis of DNA methylation

Abstract: Lopez, C. M. R., Croxford, A. E., Wilkinson, M. J. (2008). High-resolution melt analysis for SNP discovery, linkage mapping and analysis of DNA methylation Comparative Biochemistry and Physiology A-Molecular & Integrative Physiology, Abstracts, Annual Meeting of the Society-for-Experimental-Biology, Marseille, FRANCE, JUL 06-10, 2008, 150, (3), S49-S50

Detection of methylation in nucleic acid sequence

Abstract: The present invention provides a method for detecting and/or quantifying the presence of, and relative abundance of, methylated nucleic acid bases within double- stranded nucleic acid by i) contacting a double-stranded nucleic acid sample with an intercalating fluorescent dye when bound to the nucleic acid sample fluoresces when exposed to light of a wavelength capable of causing the dye to fluoresce; 2) altering the hybridisation conditions of the solution containing the double-stranded nucleic acid-dye complex such that dissociation of the two strands of the said nucleic acid-dye complex occurs at a rate that permits progressive release of the dye 3) and monitoring the difference in fluorescence, and uses thereof.

Epigenetic control of stomatal number in response to humidity environment

Abstract: Tricker, P., Gibbings, G., Cryer, N., Hadley, P., Wilkinson, M. J. (2008). Epigenetic control of stomatal number in response to humidity environment. Comparative Biochemistry and Physiology A - Molecular & Integrative Physiology 150, (3), S190-S191. Proceedings of the Annual Main Meeting of the Society of Experimental Biology, Marseille, France, 6-10 July 2008. IMPF: 01.71 RONO: 00