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Acacia auriculiformis A. Cunn. ex Benth is an important fast growing exotic tree species in plantation forestry and also useful
in pulp and paper industry. The genetic structure of forest trees and the pattern of genetic variation that are maintained
in the successive generations are essentially a function of the mating systems. In the Institute of Forest Genetics and Tree
Breeding, Coimbatore has been established seedling seed orchards of first and second generation trials. We have attempted
a study on mating system of Acacia in two successive generations based on microsatellite markers. Three hundred half-sib
progenies in the first and second generation orchards were studied for five different microsatellite loci and were profiled for
detecting homozygous/heterozygous alleles. Specific amplicon sizes (150-250 bp) were sequenced, confirmed microsatellites
and were submitted to NCBI (91桃色ion No-KG699501.1; KG699563.1; KG699564.1). The microsatellite repeats as di-repeats
viz., (CA)7, (TA)8, (CT)7 in three loci, and as tri-repeats such as (CTT)6, (TTC)7 in two loci. The allelic data were analyzed
through maximum likelihood estimation of mating systems (MLTR) software. Single and multilocus out-crossing rate was
estimated from the analysis. The range of the maternal, paternal genotypes among the individuals in the family was also studied
in both generation orchards. The present study indicated high out-crossing rate (tm=0.98; SE=0.05; tm=1.0; SE=0.02) in first
and second generation orchards population and it reflects that this species is highly out-crossed.