A new DNA marker CafLess-TCS1 for selection of caffeine-less tea plants

A clonal line of Camellia taliensis, 'Taliensis-akeme' has a recessive caffeine-less gene. To accelerate breeding of caffeine-less tea cultivars using this gene, DNA markers are indispensable for selecting heterozygotes that do not show a caffeine-less phenotype as parental lines. Therefore, we tried to determine the sequence of the six tea caffeine synthase (TCS) genes to search for polymorphisms and to prepare one of the TCS genes as a selection marker. Six TCS genes and the caffeine-less trait were mapped on the reference linkage map of tea. Strong linkage between the caffeine-less phenotype and TCS1 indicate that it is a promising candidate as a causative gene of the caffeine-less trait. We decided to use a three-nucleotide insertion in TCS1 that can be distinguished by sequencing as a selection marker named 'CafLess-TCS1'. Caffeine-less individuals appeared in the progeny population of caffeine-less heterozygous individuals selected using 'CafLess-TCS1'. These results confirmed that the developed 'CafLess-TCS1' will be an effective selection marker for breeding of caffeine-less tea cultivars.