New Citrus chloroplast haplotypes revealed by molecular markers using Algerian and Spanish accessions

Thirty-seven chloroplast molecular markers were used to evaluate the genetic diversity and infer the phylogenetic relationship of 24 Algerian Citrus accessions from the Institut Technique de l′Arboriculture Fruitiere et de la Vigne germplasm bank. The reliability and consistence of the clustering distribution was further asserted including 5 Spanish accessions from the Instituto Valenciano de Investigaciones Agrarias. The accessions were positioned on a phylogenetic tree of the genus Citrus based on previous analyses of the whole sequence of citrus chloroplast. Algerian accessions clustered into two main clades mostly differentiated by the occurrence of either mandarin or pummelo chloroplast types. All 7 mandarins analyzed were grouped in the same clade while the other cluster subdivided in 4 groups, included 1 lumia, 3 lemons, 2 grapefruits and 11 sweet oranges. Algerian grapefruit accessions were grouped together with the pummelos in a single cluster while all sweet oranges formed an independent and homogenous clade. Interestingly, the lemons studied were clustered in 3 different subclusters while Citrus lumia genotype was isolated in a different group. These results suggest that in contrast to the studied Algerian mandarins or sweet oranges, that share all the same mandarin or sweet orange chloroplast haplotype, the high diversity of current lemon accessions is at least partially correlated with the identity of different pummelo progenitors which evolved from a common ancestor. In addition, the data indicate that Citrus lumia is a new type of citrus chloroplast that appears to be phylogenetically related to the chloroplasts of the pummelo and micrantha group.