Full Length Research Paper

Identification of AFLP markers linked with cocoon weight genes in silkworm (Bombyx mori L.)

S. Z. Mirhosseini¹, A. R. Bizhannia²*, B. Rabiei³, M. Taeb² and A. R. Siedavi⁴

¹Department of Animal Science, Guilan University, Rasht, Iran.

²Department of Agricultural Biotechnology, Islamic Azad University, Science and Research Branch, Tehran, Iran.

³Department of Plant Breeding, Guilan University, Rasht, Iran.

⁴Animal science Department, Islamic Azad University, Rasht Branch, Rasht, Iran.

*Corresponding author. E-mail: alirezabizhannia@yahoo.com.

Abbreviations: QTLs, Quantitative traits loci; RFLP, restriction fragment length polymorphism; AFLP, amplified fragment length polymorphism; PCR, polymerase chain reaction; CIM, composite interval mapping; LOD, logarithmic of odds score; LRS, likelihood ratio statistic; dNTPs, deoxynucleoside 5’-triphosphate.

Accepted 1 February, 2010

   Abstract

DNA markers used in assisting selection method is a safe method in breeding process, due to deletion of environmental conditions, and it is an important tool in preparing linkage map and QTLs mapping. In mulberry silkworm that is, foundation of world sericulture, its major production- economic characteristics are polygenic. In this study, we want to determine QTL(s) affecting cocoon weight trait by AFLP markers. For this reason, we used 20 selected primer combinations from among 81 primers combinations of PstI/TaqI at the level of three F2 populations including 33, 36 and 34 offsprings sample, respectively. These populations were obtained by crossing two lines of Lemon Khorasan (as maternal) and 107 (as paternal). The parental lines, F1 and F2 individuals’ DNA were extracted with phenol-chloroform method. Then they were digested by two restriction enzymes (TaqI and PstI) and amplified by using of appropriate adaptors. These amplified samples are transfered on annealed 6% polyacrylamide gels. After genotyping of individuals, the linkage maps of populations were drawn by Map manager/QTX and QTL Cartographer ver.2.5 softwares. Number of total and polymorphic bands that formed to 20 primer combinations in each populations were 930, 944, 810 and 142, 171, 178 bands, respectively. Therefore polymorphic frequencies were 15.27, 18.11 and 21.97%. The obtained linkage maps were included in 16, 18 and 24 linkage groups. The total length of this linkage maps and average distance between two markers were 2186.40, 2582.50 and 2392.60 cM, and 18.37, 16.45 and 14.95 cM, respectively. The detection of QTLs numbers of cocoon weight character in each F2 populations also showed 1, 6 and 1 loci in LRS>17 (LOD > 3.7) by compound interval mapping methods, respectively.

Key words: Silkworm, QTL analysis, linkage map, molecular markers and AFLP.