St. augustinegrass (Stenotaphrum sp.) is a warm-season perennial turfgrass that grows widely in tropical regions around the world. St. augustinegrass is valued for both its turf performance and high levels of resistance to biotic and abiotic stresses. The current study was aimed at developing nuclear microsatellite markers for st. augustinegrass. Pyrosequencing of an enriched microsatellite library on the Roche FLX platform using a 454 Titanium kit produced 57,306 sequence reads; 2614 of which contained short tandem repeats. One hundred primer pairs were tested with 18 accessions from the U.S. Department of Agriculture National Plant Germplasm System st. augustinegrass collection grown in Griffin, GA. This collection contains both Stenotaphrum dimidiatum and Stenotaphrum secundatum accessions. Among revealed 100 primer pairs, 33 were polymorphic. A total of 175 alleles were amplified. The number of observed alleles per primer pair ranged from two to 10, with an average of 5.3. Shannon’s information index and Nei’s genetic diversity values were 0.4403 and 0.2873, respectively. This set of microsatellite markers is useful for assessment of genetic diversity and construction of molecular genetic linkage maps in st. augustinegrass.
Zhiyong Wang, Paul Raymer and Zhenbang Chen
Xiaoli Wang, Zhiyong Wang, Li Liao, Xinyi Zhang and Changjun Bai
Carpetgrass [Axonopus compressus (Sw.) Beauv.] is an important warm-season perennial turfgrass that is widely used in tropical and subtropical areas. The genetic diversity of 63 carpetgrass accessions in China was studied using simple sequence repeat (SSR) markers. Fourteen SSR primer combinations generated a total of 49 distinct bands, 48 (97.96%) of which were polymorphic. The number of observed alleles ranged from 2 to 6, with an average of 3.5. Coefficients of genetic similarity among the accessions ranged from 0.24 to 0.98. Unweighted pair-group method with arithmetic means (UPGMA) clustered the 63 accessions into three groups, and not all samples from the same region belonged to the same group. SSR markers will promote marker-assisted breeding and the assessment of genetic diversity in wild germplasm resources of carpetgrass.
Xinyi Zhang, Li Liao, Yang Liu, Zhiyong Wang and Jianxiu Liu
Chrysopogon aciculatus (Retz.) Trin. is a perennial turfgrass for its low management and resistance. To develop simple sequence repeat (SSR) markers for C. aciculatus, we used four Roche 454 pyrosequencing, combined with the magnetic bead enrichment method FIASCO (fast isolation by amplified fragment length polymorphism of sequences containing repeats) to isolate from the C. aciculatus. A total of 66,198 raw sequencing reads were obtained with 4289 sequences (6.48%) were fit for primer pair design. One hundred microsatellite loci were selected to test the primer amplification efficiency in 20 accessions, and out of these, 11 loci were polymorphic. The amount of observed alleles ranged from three to six, with an average of 3.64. Nei’s genetic diversity values ranged from 0.085 to 0.493, with an average of 0.293. Shannon’s information index values ranged from 0.141 to 0.686, with an average of 0.428. Twenty accessions were clustered into three groups by unweighted pair-group method with arithmetic means (UPGMA). These SSR markers will provide an ideal marker system to assist with gene targeting, cultivar variety or species identification, and marker-assisted selection in C. aciculatus species.
Xinyi Zhang, Li Liao, Zhiyong Wang, Changjun Bai and Jianxiu Liu
Molecular genetic diversity and relationships among 86 Chrysopogon aciculatus (Retz.) Trin. accessions were assessed using intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers. Twenty-five ISSR markers generated 283 amplification bands, of which 266 were polymorphic. In addition, 576 polymorphic bands were detected from 627 bands amplified using 30 SRAP primers. Both marker types revealed a high level of genetic diversity, with ISSR markers showing a higher proportion of polymorphic loci (PPL; 94%) than SRAP markers (91.87%). The ISSR and SRAP data were significantly correlated (r = 0.8023). Cluster analysis of the separate ISSR and SRAP data sets clustered the accessions into three groups, which generally were consistent with geographic provenance. Cluster analysis of the combined ISSR and SRAP data set revealed four major groups similar to those based solely on ISSR or SRAP markers. The findings demonstrate that ISSR and SRAP markers are reliable and effective tools for analysis of genetic diversity in C. aciculatus.