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.
Yan Liu, Hailin Guo, Yi Wang, Jingang Shi, Dandan Li, Zhiyong Wang, and Jianxiu Liu
Seashore paspalum (Paspalum vaginatum) is a notable warm-season turfgrass. Certain germplasm resources are distributed in the southern regions of China. The objectives of this study were to investigate the genetic diversity and genetic variation of Chinese seashore paspalum resources. Morphological characteristics and sequence-related amplified polymorphism (SRAP) markers were used to assess genetic relationships and genetic variation among 36 germplasm resources from China and six cultivars from the United States. The results showed significant variation for 13 morphological characteristics among 42 tested seashore paspalum accessions, and that the phenotypic cv was, in turn, turf height > turf density > internode length > inflorescence density > leaf width > reproductive branch height > spikelet width > leaf length > spikelet number > inflorescence length > internode diameter > inflorescence width > spikelet length. According to the morphological characteristics and cluster analysis, 42 seashore paspalum accessions were divided into six morphological types. In total, 374 clear bands were amplified using 30 SRAP primer combinations; among these bands, 321 were polymorphic with 85.83% polymorphism. SRAP marker cluster analysis showed that 42 seashore paspalum accessions were grouped into seven major groups, with a genetic similarity coefficient ranging from 0.4385 to 0.9893 and genetic distance values ranging from 0.0108 to 0.8244. The high level of genetic diversity occurred among Chinese germplasm, and the genetic distance was relatively high between Chinese germplasm and cultivars introduced from the United States. The patterns in morphological trait variations and genetic diversity will be useful for the further exploitation and use of Chinese seashore paspalum resources.
Qing Shen, Hua Bian, Hai-yan Wei, Li Liao, Zhi-yong Wang, Xiao-yan Luo, Xi-peng Ding, Zhenbang Chen, and Paul Raymer
Seashore paspalum (Paspalum vaginatum) is an important warm-season turfgrass distributed in tropical and coastal areas. It has excellent resistance to abiotic stresses, such as salinity, drought, and low temperature. However, the research on genetic diversity of local P. vaginatum collections from China is limited. In this study, the genetic diversity among 58 P. vaginatum accessions from four different provinces in China and four cultivars were assessed using simple sequence repeat (SSR) markers. The results indicated that a total of 45 alleles were detected by 19 polymorphic markers, with a range of 2 to 4 and an average of 2.4 alleles per marker. The genetic similarity coefficients between each pair of the 58 P. vaginatum accessions and four cultivars ranged from 0.51 to 1.00, with an average of 0.77. The range of variation of Shannon diversity index of each SSR marker was 0.047 to 1.075, with an average of 0.486. The polymorphic information content of each SSR marker varies from 0.016 to 0.577, with an average of 0.249. The results of cluster analysis and principal component analysis (PCA) showed that 58 P. vaginatum accessions and four cultivars were divided into four groups. These results provide the theoretical basis for the genetic diversity assessments and molecular marker–assisted breeding of P. vaginatum species.