Three non-pollinating chalcid fig-wasp species have been reported from a dioecious fig, Ficus hirta Vahl., Philotrypesis josephib (tribe Philotrypesini), Sycoscapter hirticola and Sycoryctes sp. (tribe Sycoryctini). They oviposit into ovaries containing Valisia larvae by inserting their long ovipositor through the fig wall. Microsatellites across 24 geographical sites for these two tribes were collected to establish species genetic diversity, inbreeding and species boundaries. We sampled representatives of each fig wasp species emerging from individual figs at each location. For Philotrypesis, 262 individuals, each from a different fig, were genotyped at 6 unlinked microsatellite loci. The genotype data of Philotrypesis is in the sheet 1 with detailed sampling information and nuclear locus. For Sycoryctini, 203 individuals, each from a different fig, were genotyped at 7 unlinked microsatellite loci. The genotype data of Sycoryctini is in the sheet 2 with detailed sampling information and nuclear locus.

Samples were collected from South China to Java (Supporting Information Table S1). In each location, 10-30 mature figs were collected just before wasp emergence. The figs were placed in fine-mesh bags to allow the wasps to emerge naturally. The wasps that emerged were preserved in 95% ethanol and stored at -20°C until DNA extraction.

For microsatellite data, a single female wasp per fig was genotyped for both Philotrypesis and Sycoryctini. For Philotrypesis, 262 individuals were genotyped at 6 unlinked microsatellite loci (P50, P86, P91, P123, P129 and P202) that had been developed for F. hirta Philotrypesis [54]. For Sycoscapter-Sycoryctes, 203 individuals were genotyped at 7 unlinked microsatellite loci (S7, S27, S52, S122, S132, S137 and S162) that had been developed for F. hirta Sycoscapter [55]. Molecular techniques followed Abellò et al. [54] and Ahanchédé et al. [55].

Method of microsatellite amplification for Sycorictini and Philotrypesis were the same expect for the annealing temperature. The amplification reactions were conducted using a PTC-200 thermal cycler (Bio-Rad, Hercules, CA) in 20 µl volume containing 20 ng of genomic DNA, 0.2 mM of each dNTP, 0.4 µM of fluorescent primer, 1× PCR buffer (Mg2+ free), 2.5 mM Mg2+ and 1 unit of Taq DNA polymerase (Takara, Dalian, China), using the following conditions: initial denaturation at 95°C for 5 min, 35 cycles of (94°C, 30 s; 46 to 50°C for Sycoryctini and 47 to 52 °C for Philotrypesis, 60 s; 72°C, 45 s) and a final extension of 72°C for 8 min. The fragment sizes of the PCR products were determined on the ABI PRISM 3100 Genetic Analyser (Applied Biosystems, Foster City, CA) using genotyper 4.0 and LIZ 500 (Applied Biosystems, Foster City, CA) as an internal size standard.

Microsatellite genotype data for all individuals of Philotrypesis and Syocryctini:

Sheet 1: Genotypic data of Philotrypesis

Nuclear genotypes (6 loci) for 262 individuals of Philotrypesis. Population refers to collection sites, with GPS coordinates provided in the main text. For each nuclear locus, alleles names are arbitrary (i.e., do not indicate size or sequence characteristics), and “0” indicates missing data.

Sheet 2: Genotypic data of Sycorictini

Nuclear genotypes (7 loci) for 203 individuals of Sycorictini. Population refers to collection sites, with GPS coordinates provided in the main text. For each nuclear locus, alleles names are arbitrary (i.e., do not indicate size or sequence characteristics), and “0” indicates missing data.