Island Animals populations are always with the acceleration of evolution rate and hold some special genetic characters [1], [2]. As a model species of social insects, insular honeybees attracted more attention and many Apis species or subspecies were identified and published during the past decades [3], [4], [5]. Samples from these islands were paid great attention, and new subspecies were identified and even new species evolved.

Three new species Apis koschevnikovi, Apis nigrocincta and Apis nuluensis [6], [7], [8] and some subspecies Apis cerana philippina, Apis cerana javana and Apis cerana japonica [9], [10], [11] were identified in Southeastern Asia based on morphometrics and mitochondrial DNA data. Honeybee population from Palawan were unequivocally distinct and separated from the other Philippine samples [12], [13], due to an isolation of some 200 km among them. Similarly, A. cerana population from Samui Island was distinctive from those from the Thai mainland (about 15 km between) with morphological, mitochondrial DNA and microsatellite DNA analysis [14], [15], [16], [17].

2.1. Materials

A total of 133 colonies were collected during year 2008 from Damen Island, northwest of Dongtou county in Zhejiang province, and two localities Chixi and Juxi in its adjacent mainland Cangnan county (Table 1, Fig. 1). The Damen Island covers an area of 28.7 km2, a coastline of 42.18 km, mountain area of 24.29 km2, and a small amount of flat and tidal flats. The strait between Damen Island and mainland is about 7 km as well as straight-line distance between Damen Island and Juxi, Chixi are about 94 km and 103 km, respectively. Table 1. Details of samples and sample size.

2.2. DNA extraction

Total DNA was extracted from the thorax and legs of each individual worker using animal genome DNA extraction kit (Shanghai Sangon Biological Engineering Technology & Services Co., Ltd.). Total DNA was used for both mitochondrial DNA and microsatellite analysis.

2.3. Mitochondrial DNA analysis

Mitochondrial …

3.1. Mitochondrial DNA

Ten mitochondrial haplotypes were detected among the 133 A.c. cerana individuals (NCBI Accession Number HM461260-HM461269, Table 3). There were seven haplotypes in terms of noncoding sequence, and four haplotypes in terms of coding sequence (Fig. 2). Table 3. Genetic structure of mtDNA in three locations.