An applied fingerprinting system for cultivated potato using simple sequence repeats

Abstract

The ability to quickly and accurately identify potato (Solanum tuberosum L.) clones is important to potato-breeding programs, seed and commercial potato growers, and marketing and utilization of potato cultivars. Since 1990, the Michigan State University Potato Breeding and Genetics Program has used an isozyme-based fingerprinting system to identify potato cultivars. Isozyme analysis is an economical and effective means of discriminating potato clones; however, isozyme analysis requires fresh, healthy tuber or leaf tissue. DNA-based fingerprinting using simple sequence repeats (SSRs or microsatellites) has been shown to discriminate between potato clones. The objective of this study was to identify the most useful SSR primer pairs that accurately and efficiently distinguish clones for an applied fingerprinting system of cultivated potato. SSR primer pairs with high polymorphism were selected from previous tetraploid potato studies. DNA isolated from 17 potato clones representing round-white, russet, and red market classes were visualized on both polyacrylamide (PAGE) and agarose gel systems. Polymorphism was observed in all 18 primer combinations on PAGE and 14 using agarose gel electrophoresis. All 17 cultivars were discriminated on PAGE with various combinations of two primer pairs: STIIKA using STACCAS3, STIN-HWI, or STM0031; and STACCAS3 using STGBSS1, POTM1-2, STM1104, or STM0031. The combination of STM0019, STM0031, STGBSS1, and POTM1-2 was able to differentiate all 17 clones using agarose gel electrophoresis. PAGE was determined to be the preferred system for variety identification, but agarose gel electrophoresis can be used to differentiate lines when specific varietal comparisons are needed. In addition, five different DNA source tissue types were evaluated (fresh foliar, freeze-dried foliar, fresh tuber, freeze-dried tuber epidermis, and freeze-dried tuber tissue). Amplification products were similar for all five tissue sources used for DNA isolation. This ability to isolate DNA from freeze-dried tissue will allow cultivar identification when fresh tissue is not available. The SSR primer pairs presented here can be used as a practical fingerprinting system for cultivated potato identification.