Caffeoyl CoA was detected when CGA was incubated all targets with Coenzyme A in the presence of the recombinant protein, whereas no metabolic product was detected from cultures carrying an empty plasmid. Real time PCR In order to assess the involvement in the response to UV C irradiation, the expression levels of HQT and HCT were analysed by real time PCR. Based on normalized levels, it was clear that UV C treatment induced a significant increase in transcription. Comparison between the standard curves for each enzyme revealed a correlation coefficient of 0. 98 and an effi ciency 0. 90. Linkage analysis Two single nucleotide polymorphisms were identified in the HQT and HCT parental sequences. Both parents of the mapping population were heterozygous for marker HQT snp359, that segregated in the ratio 1 2 1 in the F1 individuals, with no evidence of any segregation distortion.
This allowed the HQT gene Inhibitors,Modulators,Libraries to be placed on linkage group 5 in both the female and male maps. A fur ther 14 markers were assigned to the female LG5 four microsatellites, three S SAPs and 7 AFLPs, covering 62. 1 cM and a mean inter marker distance of 4. 4 cM. More than 70% of intervals are 4 cM in genetic length, with four gaps of 6 cM. In addition to the HQT locus, the Inhibitors,Modulators,Libraries male LG5 consists of 15 markers two SSRs one S SAP, one M AFLP and 11 AFLPs, spanning 69. 5 cM and a mean inter marker dis tance of 4. 4 cM. Seven markers were shared between the parents, allowing the alignment of their LG5. The HQT locus maps close to AFLP markers e38 m47 01 and e47 m49 06 in the female map, and to the M AFLP marker polyGA Inhibitors,Modulators,Libraries e33 02 and the microsatellite CELMS 24 in the male map.
Only the female parent was heterozygous at HCTsnp97, Inhibitors,Modulators,Libraries delivering a segregation ratio of 1 1 with no significant distortion. As a result, the HCT gene could only be located on the maternal map, where it maps to LG9, separated by 3 cM from the AFLP locus p12 m61 04 and by 8 cM from the SSAP locus cyre5 m47 02. A further six markers are present on this 58. 4 cM LG, including one SSR, two M AFLPs and three AFLPs. The marker density is 7. 3 cM, with two gaps of 10 cM. Discussion Plants synthesize a variety of secondary metabolites, which function as UV protectants, Inhibitors,Modulators,Libraries phytoalexins, flower pigments, signalling molecules and building blocks for lignin.
Some have significance in the area of human health, both as phytomedicines, which target specific health problems, and or as nutraceuticals, which provide long kinase inhibitor Wortmannin term nutritional benefit. Particular plant PPs have been associated with anti oxidant, estrogen like and vasodilatory activity, while others have proven anti inflammatory and anti cancer chemopreventive action CGA is the most widespread plant PP. Progress is being made in relation to the definition of its biosynthetic path way, with the characterisation of two acyltransferases able to synthesize p cou maroylshikimate and p coumaroyl quinate esters and a cytochrome P450 p coumaroyl ester 3 hydroxylase from a p coumaroyl ester substrate.