QTL mapping of morphological characteristics that correlated to drought tolerance in St. Augustinegrass.

St. Augustinegrass is a warm-season grass species widely utilized as turf in the southeastern U.S. It shows significant variation in plant growth and morphological characteristics, some of which are potentially associated with drought tolerance. However, the genetic basis of these variations is not well understood. Detecting quantitative trait loci (QTL) associated with morphological traits will provide a foundation for the application of genetic and molecular breeding in St. Augustinegrass. In this study, we report QTL associated with morphological traits, including leaf blade width (LW), leaf blade length (LL), canopy density (CD), and shoot growth orientation (SGO) in a St. Augustinegrass 'Raleigh' x 'Seville' mapping population containing 115 F1 hybrids. Phenotypic data were collected from one greenhouse and two field trials. Single and joint trial analyses were performed, finding significant phenotypic variance among the hybrids for all traits. Interval mapping (IM) and multiple QTL method (MQM) analysis detected seven QTL for CD, four for LL, five for LW, and two for SGO, which were distributed on linkage groups RLG1, RLG9, SLG3, SLG7, SLG8 and SLG9. In addition, three genomic regions where QTL colocalized were identified on Raleigh LG1 and Seville LG3. One genomic region on Seville LG3 overlapped with two previously reported drought-related QTL for leaf relative water content (RWC) and percent green cover (GC). Several candidate genes related to plant development and drought stress response were identified within QTL intervals. The QTL identified in this study represent a first step in identifying genes controlling morphological traits that might accelerate progress in selection of St. Augustinegrass lines with lower water usage.

Quantitative Trait Loci Associated with Gray Leaf Spot Resistance in St. Augustinegrass.

Gray leaf spot (GLS), caused by Magnaporthe grisea, is a major fungal disease of St. Augustinegrass (Stenotaphrum secundatum), causing widespread blighting of the foliage under warm, humid conditions. To identify quantitative trait loci (QTL) controlling GLS resistance, an F1 mapping population consisting of 153 hybrids was developed from crosses between cultivar Raleigh (susceptible parent) and plant introduction PI 410353 (resistant parent). Single-nucleotide polymorphism (SNP) markers generated from genotyping-by-sequencing constituted nine linkage groups for each parental linkage map. The Raleigh map consisted of 2,257 SNP markers and spanned 916.63 centimorgans (cM), while the PI 410353 map comprised 511 SNP markers and covered 804.27 cM. GLS resistance was evaluated under controlled environmental conditions with measurements of final disease incidence and lesion length. Additionally, two derived traits, area under the disease progress curve and area under the lesion expansion curve, were calculated for QTL analysis. Twenty QTL were identified as being associated with these GLS resistance traits, which explained 7.6 to 37.2% of the total phenotypic variation. Three potential GLS QTL "hotspots" were identified on two linkage groups: P2 (106.26 to 110.36 cM and 113.15 to 116.67 cM) and P5 (17.74 to 19.28 cM). The two major effect QTL glsp2.3 and glsp5.2 together reduced 20.2% of disease incidence in this study. Sequence analysis showed that two candidate genes encoding ß-1,3-glucanases were found in the intervals of two QTL, which might function in GLS resistance response. These QTL and linked markers can be potentially used to assist the transfer of GLS resistance genes to elite St. Augustinegrass breeding lines.

Detection of quantitative trait loci associated with drought tolerance in St. Augustinegrass.

St. Augustinegrass (Stenotaphrum secundatum) is a warm-season grass species commonly utilized as turf in the southeastern US. Improvement in the drought tolerance of St. Augustinegrass has significant value within the turfgrass industry. Detecting quantitative trait loci (QTL) associated with drought tolerance will allow for advanced breeding strategies to identify St. Augustinegrass germplasm with improved performance for this trait. A multi-year and multi-environment study was performed to identify QTL in a 'Raleigh' x 'Seville' mapping population segregating for phenotypic traits associated with drought tolerance. Phenotypic data was collected from a field trial and a two-year greenhouse study, which included relative water content (RWC), chlorophyll content (CHC), leaf firing (LF), leaf wilting (LW), green cover (GC) and normalized difference vegetative index (NDVI). Significant phenotypic variance was observed and a total of 70 QTL were detected for all traits. A genomic region on linkage group R6 simultaneously harbored QTL for RWC, LF and LW in different experiments. In addition, overlapping QTL for GC, LF, LW and NDVI were found on linkage groups R1, R5, R7 and S2. Sequence alignment analysis revealed several drought response genes within these regions. The QTL identified in this study have potential to be used in the future to identify genes associated with drought tolerance and for use in marker-assisted breeding.

Molecular Dissection of Quantitative Variation in Bermudagrass Hybrids (Cynodon dactylon x transvaalensis): Morphological Traits.

Bermudagrass (Cynodon (L.)) is the most important warm-season grass grown for forage or turf. It shows extensive variation in morphological characteristics and growth attributes, but the genetic basis of this variation is little understood. Detection and tagging of quantitative trait loci (QTL) affecting above-ground morphology with diagnostic DNA markers would provide a foundation for genetic and molecular breeding applications in bermudagrass. Here, we report early findings regarding genetic architecture of foliage (canopy height, HT), stolon (stolon internode length, ILEN and length of the longest stolon LLS), and leaf traits (leaf blade length, LLEN and leaf blade width, LW) in 110 F1 individuals derived from a cross between Cynodon dactylon (T89) and C. transvaalensis (T574). Separate and joint environment analyses were performed on trait data collected across two to five environments (locations, and/or years, or time), finding significant differences (P < 0.001) among the hybrid progeny for all traits. Analysis of marker-trait associations detected 74 QTL and 135 epistatic interactions. Composite interval mapping (CIM) and mixed-model CIM (MCIM) identified 32 main effect QTL (M-QTL) and 13 interacting QTL (int-QTL). Colocalization of QTL for plant morphology partially explained significant correlations among traits. M-QTL qILEN-3-2 (for ILEN; R2 = 11-19%), qLLS-7-1 (for LLS; R2 = 13-27%), qLEN-1-1 (for LLEN; R2 = 10-11%), and qLW-3-2 (for LW; R2 = 10-12%) were 'stable' across multiple environments, representing candidates for fine mapping and applied breeding applications. QTL correspondence between bermudagrass and divergent grass lineages suggests opportunities to accelerate progress by predictive breeding of bermudagrass.

High density genetic maps of St. Augustinegrass and applications to comparative genomic analysis and QTL mapping for turf quality traits.

BACKGROUND: St. Augustinegrass [Stenotaphrum secundatum (Walt.) Kuntze] is a warm-season, perennial turfgrass species well adapted for home lawns and commercial landscapes with economic and ecological value. However, a lack of genomic resources in St. Augustinegrass has hindered the full utilization of genetic variance for maximizing genetic gain and limited our understanding of the species' evolution. RESULTS: In this study, we constructed the first high-density linkage map for St. Augustinegrass using a genotyping by sequencing (GBS) approach. The integrated linkage map consists of 2871 single nucleotide polymorphism (SNP) and 81 simple sequence repeat (SSR) markers, spanning 1241.7 cM, with an average distance of 0.4 cM between markers, and thus represents the densest genetic map for St. Augustinegrass to date. Comparative genomic analysis revealed inter-chromosome arrangements and independent nested chromosome fusion events that occurred after St. Augustinegrass, foxtail millet, sorghum, and rice diverged from a common ancestor. Forty-eight candidate quantitative trait loci (QTL) were detected for turf quality-related traits, including overall turf quality, leaf texture, genetic color, and turf density. Three hot spot regions were identified on linkage groups LG3 and LG8, where multi-QTL for different traits overlapped. Several leaf development related genes were contained within these identified QTL regions. CONCLUSIONS: This study developed the first high-density genetic map and identified putative QTL related to turf quality, which provide valuable genetic resources for marker-assisted selection (MAS) in St. Augustinegrass.

Overexpression of ubiquitin-like LpHUB1 gene confers drought tolerance in perennial ryegrass.

HUB1, also known as Ubl5, is a member of the subfamily of ubiquitin-like post-translational modifiers. HUB1 exerts its role by conjugating with protein targets. The function of this protein has not been studied in plants. A HUB1 gene, LpHUB1, was identified from serial analysis of gene expression data and cloned from perennial ryegrass. The expression of this gene was reported previously to be elevated in pastures during the summer and by drought stress in climate-controlled growth chambers. Here, pasture-type and turf-type transgenic perennial ryegrass plants overexpressing LpHUB1 showed improved drought tolerance, as evidenced by improved turf quality, maintenance of turgor and increased growth. Additional analyses revealed that the transgenic plants generally displayed higher relative water content, leaf water potential, and chlorophyll content and increased photosynthetic rate when subjected to drought stress. These results suggest HUB1 may play an important role in the tolerance of perennial ryegrass to abiotic stresses.

St. Augustine grass germplasm resistant to Blissus insularis (Hemiptera: Blissidae).

St. Augustine grass (Stenotaphrum secundatum (Walter) Kuntze) is an economically important turfgrass in the southeastern United States. However, this turf species is prone to southern chinch bug, Blissus insularis Barber (Heteroptera: Blissidae) outbreaks. This insect is the most destructive pest of St. Augustine grass wherever this turf grass is grown. Host plant resistance has historically been an effective management tool for southern chinch bug. Since 1973, the 'Floratam' St. Augustine grass cultivar effectively controlled southern chinch bug in the southeast. However, southern chinch bug populations from Florida and Texas have now circumvented this resistance, through mechanisms still unknown. Therefore, identifying and deploying new cultivars with resistance to the southern chinch bug is imperative to combat this pest in an economically and environmentally sustainable manner. Currently, the number of cultivars with resistance against southern chinch bug is limited, and their efficacy, climatic adaptability, and aesthetic characters are variable. Hence, the main focus of this study is the identification of alternative sources of resistance to southern chinch bugs in previously uncharacterized St. Augustine grass plant introductions (PIs) and its closely related, crossbreeding species, Pembagrass (Stenotaphrum dimidiatum (L.) Brongniart). The PIs exhibited a wide range of responses to southern chinch bug feeding, as indicated by damage ratings. Damage ratings for seven PIs grouped with our resistant reference cultivars. Moreover, nine PIs exhibited antibiosis, based on poor development of southern chinch bug neonates, when compared with our susceptible reference cultivars. Altogether our study has produced strong support to indicate these materials are good candidates for future southern chinch bug resistance breeding in St. Augustine grass.