Construction of cDNA Library and EST Analysis Related to Seed-hair Characteristics in Carrot

Abstract. Carrot ( Daucus carota L. var. sativa ) is one of the most widely used crops in the world and is nutritionally important crop. However, seed-hair which is generated in epidermal cell of seeds causes the difficulty of the seedling process, because of the seed germination and absorption inhibitions. For these reasons, carrot seeds are commercialized after mechanical hair removal process. However, in this process, various damage and seed loss occur and breeding of hairless-seed carrot cultivar is needed to overcome these various weaknesses and additional seed production costs. In this study, cDNA libraries using 2 combinations, which were composed of short-hair seed CT-ATR 615 OP 666-13 & long-hair seed CT-ATR 615 OP 671-9, and short-hair seed CT-SMR 616 OP 659-1 & long-hair seed CT-SMR 616 OP 677-14, were constructed and EST sequences of each individuals were analyzed to reveal carrot seed-hair characteristics. Firstly, analyzed EST sequences were classified into FunCat functional categories. As a result, significant differences have been identified in metabolism category, protein folding and stabilization, protein binding, C-compound binding category from both of two combinations. Secondly, several candidate EST sequences related to seed trichome differentiation and cellulose biosynthetic process were selected based on GO data of EST sequences. These differences based on FunCat categories and candidate EST obtained by GO data analysis are thought to be involved in the formation of carrot seed hair. Finally, 741 SSR sites and 33 SNP sites were identified from analyzed EST sequences of two combinations. Then we designed SNP and SSR primer sets to develop molecular markers. These molecular markers will be used for classification of carrot cultivars and study seed-hair characteristic.

[1]  M. Sorrells,et al.  EST derived SSR markers for comparative mapping in wheat and rice , 2004, Molecular Genetics and Genomics.

[2]  P. Gupta,et al.  Transferable EST-SSR markers for the study of polymorphism and genetic diversity in bread wheat , 2003, Molecular Genetics and Genomics.

[3]  Shim EunJo,et al.  Analysis of Seed Hair Formation Related Genes by EST Profiling in Carrot (Daucus carota var. sativa) , 2010 .

[4]  P. Langridge,et al.  Interspecific transferability and comparative mapping of barley EST-SSR markers in wheat, rye and rice , 2005 .

[5]  Oh GyuDong,et al.  EST Profiling for Seed-hair Characteristic and Development of EST-SSR and SNP Markers in Carrot , 2010 .

[6]  Development of SNP Molecular Markers Related to Seed-hair Characteristic Based on EST Sequences in Carrot , 2013 .

[7]  D. Hamer,et al.  High-throughput SNP genotyping by allele-specific PCR with universal energy-transfer-labeled primers. , 2001, Genome research.

[8]  Chuanping Yang,et al.  Generation and analysis of expressed sequence tags from a cDNA library of Tamarix androssowii , 2006 .

[9]  M. Sorrells,et al.  Data mining for simple sequence repeats in expressed sequence tags from barley, maize, rice, sorghum and wheat , 2002, Plant Molecular Biology.

[10]  J. L. Weber,et al.  Survey of plant short tandem DNA repeats , 1994, Theoretical and Applied Genetics.

[11]  A. Kerlavage,et al.  Complementary DNA sequencing: expressed sequence tags and human genome project , 1991, Science.

[12]  W. Powell,et al.  Polymorphism revealed by simple sequence repeats , 1996 .

[13]  H. Mewes,et al.  The FunCat, a functional annotation scheme for systematic classification of proteins from whole genomes. , 2004, Nucleic acids research.

[14]  M. Ashburner,et al.  Gene Ontology: tool for the unification of biology , 2000, Nature Genetics.

[15]  Dmitrij Frishman,et al.  MIPS: analysis and annotation of proteins from whole genomes in 2005 , 2005, Nucleic Acids Res..

[16]  Ling-Jian Wang,et al.  Gene expression and metabolite profiles of cotton fiber during cell elongation and secondary cell wall synthesis , 2007, Cell Research.

[17]  K. O. Ayeh Expressed sequence tags (ESTs) and single nucleotide polymorphisms (SNPs): Emerging molecular marker tools for improving agronomic traits in plant biotechnology , 2008 .

[18]  A. Walsh,et al.  Mining single-nucleotide polymorphisms from hexaploid wheat ESTs. , 2003, Genome.

[19]  I. Day,et al.  An efficient procedure for genotyping single nucleotide polymorphisms. , 2001, Nucleic acids research.