Abstract
Improving mulberry leaf production with enhanced leaf quality holds the key to sustain the ever increasing demand for silk. Adoption of modern genomic approaches for crop improvement is severely constrained by the lack of sufficient molecular markers in mulberry. Here, we report development and validation of 206 EST derived SSR markers using transcriptome data generated from leaf tissue of a drought tolerant mulberry genotype, Dudia white. Analysis of transcriptome data containing 10169 EST sequences, revealed 1469 sequences with microsatellite repeat motifs. We designed a total of 264 primers to the most appropriate repeat regions, of which 206 were locus specific. These markers were validated with 25 diverse mulberry accessions and their transferability to closely related species belonging to family Moraceae was examined. Of these markers, 189 revealed polymorphism with up to 8 allelic forms across mulberry species, genotypes and varieties with a mean of 3.5 alleles per locus. The markers also revealed higher polymorphic information content of 0.824 among the accessions. These markers effectively segregated the species and genotypes and hence, can be used for both diversity analysis and in breeding applications. Around 40% of these markers were transferable to other closely related species. Along with the other genic and genomic markers, we report a set of over 750 co-dominant markers. Using these markers we constructed the first genetic linkage map of mulberry exclusively with co-dominant markers.
Highlights
India presently is the second largest producer of raw silk next to China [1]
The cDNA library developed from drought stressed leaf tissues of mulberry genotype, Dudia white, contains 10190 Expressed Sequence Tags (ESTs) (NCBI-Sequence Read Archive (SRA) SRP047446)
The ESTs were searched at National Center for Biotechnology Information (NCBI) against nr database with a stringent E-value and 10169 EST sequences were selected and analyzed for sequence redundancy
Summary
India presently is the second largest producer of raw silk next to China [1]. The total raw silk produced in India (23000 MT) is far below that produced in China [2][1]. Development and Validation of EST SSR Markers in Mulberry (Bombyx mori L.) is predominantly reared in India. Though multivoltine races are resilient to tropical climates, cocoon production efficiency is significantly lower than the bivoltine races reared in China [1]. The second and the most important constraint is the low production of mulberry (Morus sp.) leaf predominantly due to insufficient water availability. Efforts are being made to improve the crop performance under water limiting conditions. Because of the perennial and outbreeding nature of mulberry, conventional breeding for a focused crop improvement has been a challenge
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