Poa annua L. has long been a cultivated weed on golf courses. However, the recent development of improved cultivars of creeping bluegrass (Poa annua reptans Hausskn.) has generated an increasing interest in selection and breeding of this species. Inter-simple sequence repeat (ISSR) PCR is a relatively new method for genotype identification and measuring genetic diversity and was employed in this study for differentiating among creeping bluegrass genotypes. The objectives of this study were to test ISSR primers for production of polymorphic fragments and ascertain the applicability of ISSR PCR to distinguish closely related genotypes. Eight primers produced fragments, of which 77.3% were polymorphic, and primers UBC849, UBC850, and UMN001 produced over 75% of the total polymorphic fragments. These three primers had sufficient resolution to distinguish all but two of the diploid creeping bluegrass accessions. This method was a simple, fast, and relatively inexpensive method to produce useful DNA fragments in creeping bluegrass. It is a robust method for detecting polymorphic loci that can be used in the study of genetic relatedness, heritability, or linkage to important traits, development of linkage maps, and marker-assisted breeding.
An inexpensive system for maintaining desired water potentials throughout seed germination was developed. During hydration, a water reservoir at the base of inclined petri dishes allowed continual saturation of filter paper on which seeds were placed. During dehydration, seeds were exposed to equilibrium vapor pressures above saturated salt solutions. Constant temperature, necessary to prevent condensation of water vapor, was achieved via a small (0.2 A) fan that furnished and circulated heat throughout an insulated chamber in which salt solutions were placed. By operating the chamber above ambient laboratory temperature, interior cooling was not required. The system allowed manipulation of the rate, degree, and frequency of dehydration episodes to which germinating seeds were exposed.