Microsatellite markers developed by next-generation sequencing differentiate inbred lines of Apis melliferaJournal of Asia-Pacific Entomology


Hye-Kyung Kim, Iksoo Kim, Myeong-Lyeol Lee, Yong-Soo Choi, Byung Rae Jin
Insect Science


Development of microsatellite markers for Astrotrichilia asterotricha (Meliaceae), an endemic tree in Madagascar, using next-generation sequencing technology

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Development of Microsatellite Primers in the Protected Species Viola elatior (violaceae) Using Next-Generation Sequencing

Mélina Celik, Jérôme Wegnez, Chantal Griveau, Josie Lambourdière, Jose Utge, Florence Noël, Jawad Abdelkrim, Nathalie Machon

Making next-generation sequencing work for you: approaches and practical considerations for marker development and phylogenetics

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Apis mellifera: Honeybee Ecology.



ge on , R.D f Kor publ

Apis mellifera


Inbred bees s of lly a h m ix in 0,580 repeat motifs were identified. Di-nucleotide repeats, comprised mostly of t indus et al., 2005; Viuda-Martos et al., 2008). However, bee colonies are vulner- graphic distribution (Weber et al., 1991; Blair and McCouch, 1997; Kim ensity data set. In this markers suitable for mellifera. These microe and maintenance of

Journal of Asia-Pacific Entomology 18 (2015) 801?805

Contents lists available at ScienceDirect

Journal of Asia-Pac .ebreeding (Bigio et al., 2014). For several years, we have been breeding

Apis mellifera at the National Academy of Agricultural Science, a government institute in Korea (unpublished), by artificial insemination. the valuable inbred lines of A.melliferamaintained in Korea.

Materials and methodstask allocation and colony fitness (Oldroyd and Fewell, 2008). From the breeding perspective, however, this behavior makes selection of desired traits difficult, because several male traits are simultaneously passed to offspring. Thus, artificial insemination is used as an alternative to control fraction of the genome generates a large, high-d study, we used NGS to identify microsatellite genotyping and identifying six inbred lines of A. satellite markers will be important for the tracable to colony collapse disorder, pests, and disease. Thus, breeding new strains of resistant bees might be essential for sustainable beekeeping.

Queen honey bees mate with two ormoremales (Adams et al., 1977;

Laidlaw and Page, 1985), a behavior that increases colony productivity and resistance to disease, parasites, and pathogens, resulting in enhanced et al., 2010, 2012, 2014). Several techniques are now available to enrich microsatellite libraries (Zane et al., 2002) and streamline the process for screening and selecting suitable microsatellite markers. Of these, nextgeneration sequencing (NGS) is fast and cost effective (Gardner et al., 2011). In this approach, high-efficiency random sequencing of a smallMicrosatellites, a special class of repetitive D found in both coding andnoncoding regions in ? Corresponding author. Tel./fax: +82 51 200 7594.

E-mail address: brjin@dau.ac.kr (B.R. Jin). http://dx.doi.org/10.1016/j.aspen.2015.10.003 1226-8615/? 2015 Korean Society of Applied Entomoloyal jelly, pollen, propolis,

Calderone, 2000; Jensen iations are widely used as codominant multi-allelic markers in genome mapping and analysis of parentage, population structure, and phylogeo-lination and for producing honeybee venom, ro and other products (Burdock, 1998; Morse andNext-generation sequencing


Honeybees, genus Apis, are importanloci with high repeat numbers and were covered by overlapping contigs. PCR amplification yielded clean fragments from 34 of these loci, and nine were selected to differentiate the inbred lines. Genotyping at these nine loci revealed 4?14 alleles per locus, with average 10.11, as well as 0.1875?0.9592 per-locus observed heterozygosity, and 0.5432?0.8669 per-locus polymorphic information content. Thus, some of these loci are profoundly variable. Phylogenetic analysis using the nine microsatellite loci separated individual honey bees from different inbred lines verywell, suggesting that thesemarkersmay be used to trace the six inbred lines of A.melliferamaintained in Korea. ? 2015 Korean Society of Applied Entomology, Taiwan Entomological Society and Malaysian Plant Protection

Society. Published by Elsevier B.V. All rights reserved. trial insects used for polsome prokaryotic genomes (Tautz and Renz, 1984). These loci consist of motifs (e.g., di-, tri-, or higher-order nucleotidemotifs) variably repeated in tandem, and generate a profusion of distinguishable alleles. These var-Keywords:

Microsatellite [AT]n motifs, were two-fold more common than tri-nucleotide repeats. We arbitrarily selected 50 microsatelliteAvailable online 17 October 2015

Science in Korea. In total, 2Microsatellite markers developed by nextdifferentiate inbred lines of Apis mellifera

Hye-Kyung Kim a,b, Iksoo Kim c, Myeong-Lyeol Lee a, Y a Department of Agricultural Biology, National Institute of Agricultural Science and Technology b College of Natural Resources and Life Science, Dong-A University, Busan 604-714, Republic o c College of Agricultural and Life Science, Chonnam National University, Gwangju 500-757, Re a b s t r a c ta r t i c l e i n f o

Article history:

Received 9 September 2015

Revised 2 October 2015

Accepted 11 October 2015

Microsatellites, a special clas netic polymorphism, especia line the identification of suc nucleotide sequences from s j ourna l homepage: wwwNA sequences, have been all eukaryotic nuclear and gy, Taiwan Entomological Society anneration sequencing g-Soo Choi a, Byung Rae Jin b,? .A., Wanju-gun 565-850, Republic of Korea ea ic of Korea repetitive DNA sequences, have become one of the most popular markers of ges next-generation sequencing (NGS)methods have become available to streamarkers. Based on 45,476 contigs generated by NGS, we obtained 205 Mbp of bred lines of Apis melliferamaintained at the National Academy of Agricultural ific Entomology l sev ie r .com/ locate / japeHoneybee samples

As a part of a breeding project, six Apis mellifera inbred lines (A, C, D,

E, F, and G) were generated and maintained for several years via d Malaysian Plant Protection Society. Published by Elsevier B.V. All rights reserved. artificial insemination (Fig. 1). The breeding scheme and characterization of each line will be published in detail elsewhere. Lines A, C, F, and G were bred from A. mellifera ligustica collected in Australia, northeastern China, United States, and Korea, respectively. The A line is dark brown and not aggressive, but susceptible to disease and has low fertility,while the C line is light brown, highly fertile, and produces royal jelly honey abundantly. The brown-colored F line has superior fertility and hive splitting, but consumes more feed. Similarly, the brown-colored