Abstract
A simple procedure was developed to convertLathyrus sativus defence-related expressed sequence tags (ESTs) into mappable genetic markers by using PCR. Twenty-nine STS primer pairs were generated on the basis of sequence information from anL. sativus cDNA library. These primers were used to screen for polymorphisms between 2L. sativus accessions, ATC 80878 and ATC 80407, resistant and susceptible, respectively, toMycosphaerella pinodes infection. All 29 primer pairs amplified PCR products in both accessions, 11 of which amplified multiple RAPD-like products. The remaining 18 primer pairs amplified single monomorphic products. Following cloning, sequencing, and database searches, 17 of 18 PCR products were confirmed to have amplified the targeted genome region. Ten of these 17 STS primer pairs revealed polymorphisms between ATC 80878 and ATC 80407 when PCR products were digested with a range of restriction endonucleases. These results suggest that the STS-based PCR analysis will be useful for generating informative molecular markers inL. sativus for future genome mapping experiments.
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Abbreviations
- CAB:
-
chlorophyll a/b binding
- CAPS:
-
cleaved amplified polymorphic sequence
- EST:
-
expressed sequence tag
- SSR:
-
simple sequence repeat
- STS:
-
sequence-tagged sites
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Skiba, B., Ford, R. & Pang, E.C.K. Amplification and detection of polymorphic sequence-tagged sites inLathyrus sativus . Plant Mol Biol Rep 21, 391–404 (2003). https://doi.org/10.1007/BF02772588
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DOI: https://doi.org/10.1007/BF02772588