Developing summerdormant tall fescue for the southern Great Plains

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1 Developing summerdormant tall fescue for the southern Great Plains

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3 Persistence is the major constraint of growing tall fescue in south-central USA 40-60% stand loss in a year Improve persistence Drought tolerance Summer dormancy

4 Three distinct morphotypes of tall fescue

5 Mediterranean Tall Fescue Contains high levels of summer dormancy Summer

6 Key environmental factors for Summer dormancy?

7 Genotypes and conditions evaluated Three genotypes with known phenotype: Summer active (SA) Strongly summer dormant (SSD) Moderately summer dormant (MSD) Daylength and temperature Short day (SD, 10 h) Long day (LD, 16 h) High temperature (HT, 34 C) Optimum temp. (OT, 24 C) Moisture and vernalization Well watered (+W) Water stressed (-W) Vernalized (+V) Non-vernalized (-V) Three genotypes in same pot

8 Growth chamber conditions Chamber 1 Long day, high temperature Chamber 2 Short day, high temperature +V -V +V -V -W +W -V +V -V +V +W -W Chamber 3 Chamber 4 Long day, optimum temperature Short day, optimum temperature +V -V +V -V -W +W -V +V -V +V -W +W

9 Water stress in optimum temperature conditions Without water With water SD+OT -W-V SD+OT -W+V SD+OT +W-V SD+OT +W+V Long day and -W SA MSD SSD MSD SA SSD Water is not a critical factor for summer dormancy Before top watering After top watering

10 Key factor for dormancy induction Dormancy response could be phenotyped by comparing growth of plants under optimal (SD, OT,+W, and +V) versus dormancy inducing (LD, OT, +W, and +V) conditions Dormant genotypes grew significantly less than the active for: - number of tillers (5.6 vs. 15.8) - leaf elongation (44.3 vs days) - fresh weight (6.0 vs g) - dry weight (3.1 vs. 8.8 g) under long day, optimum temperature, +W, and +V conditions

11 Validation of the concept in a segregating population Fresh wt. Dry wt R43-64 Pop. Max Pop. Min Distinct parental growth (reduced growth of dormant parent under LD, OT, +W, and +V) Transgressive segregation in the population

12 Marker development Similar plant characteristics Difficult to distinguish morphologically Nuclear and chloroplast genome specific markers used for identification Sequences of chloroplast specific markers showed additions, deletions and substitutions

13 Mediterranean Continental N10 L4 B4 O5 E1 C3 B11 M11 M2 D2 N8 Morphotype specific chloroplast primers Four primer pairs identified which can clearly distinguish the Continental and Mediterranean morphotypes Primer NFTCHL045 has a 47 bp deletion in the Continental materials and can be differentiated in agarose gel NFTCHL045 was used to classify tall fescue collection Samples 47bp deletion in DNA sequence DNA from all tall fescue plants collected from a field at Ardmore, OK posses the 47 bp deletion fragment correspond to Continental morphotype Noble Foundation Forage Analysis core facility included the NFTCHL045 primer assay in their services

14 Chloroplast genome based phylogeny 3.4 PI289654_F Tetraploid fescue PI289651_F Torpedo Rhizomatous Summer active PDF_584 Continental KY-31 Flecha_Max_Q Summer dormant Resolute Mediterranean Prosper PI283313_F Festuca mairei PI283312_F Rita Meadow fescue Ensign Nucleotide Substitutions (x100) (x100) Very similar relationships among the Fescue grasses were obtained from the SSR-STS marker analysis 20 0

15 Genetic loci controlling summer dormancy

16 Development of a mapping population Parental selection Flecha MaxQ Dormant plants selected from Flecha field Re-grown in greenhouse Selected most dormant plant Bi-parental crossing R43-64, parent of another population selected as the summer active parent

17 Construction of parental linkage maps Mapping population was genotyped with SSR, STS, and DArT markers Parental maps were constructed following CP model of JoinMap nffg nffg nffg nffg nffg nffg nffg nffg nffg nffg nffa nffa lopt nffa nffa nffa lopt nffa nffg nffg cnl nffa lopt nffa nffa lopt nffa nffg nffa nffa lopt nfmf nfmf nffa nfmf nfmf nfmf lopt lopt nffa lopt lopt np nfmf lopt ctg ctg nffa nffa nffa nffg nffg nfmf lopt lopt nffa nffg nffa nffa nffa nffa nfmf nfmf nffg nffa nffa nfmf nfmf lopt lopt nffa nffa nffa nffa nffa nffa nffg nffg lopt nfmf nffa nfmf nfmf nfmf lopt nfmf lopt lopt nffa lopt nffa lopt np nffa nffa nffa nffa nffa lopt nffg lopt nfmf nffa nffa nffg lopt nffa lopt nffa lopt lopt nffg nffg lopt lopt nffa lopt lopt nffg lopt lopt np nffa lopt nffa lopt np lopt nffa lopt np nffg nffa nffa lopt nffa nfmf cnl lopt ctg nffa nfmf lopt lopt lopt nffg lopt lopt nffg lopt lopt nffg lopt nffg nffg lopt nffa lopt np lopt lopt nffa nffa lopt lopt nffa R43-64 map lopt np lopt lopt nffa nffa nffa nffa nffa nffa nffg nffg nffg nffg nffg nffg nffg nffg nffg nfmf nffa nffg lopt lopt lopt lopt lopt lopt np nffa lopt lopt nffg nffg lopt np lopt ctg nffg nffa nffa nffg nffg lopt np nffa nffa nffa nffg R43-64 map: length 1509 cm, total loci 541, density 2.79 cm/marker map: length 1044 cm, total loci 141, density 7.41 cm/marker

18 Field evaluation of NFTS06 population 208 genotypes and parental ramets evaluated in replicated trials at two locations Segregation for senescence can be observed in extreme weather conditions

19 Plant height (cm) Frequency Phenotyping summer dormant mapping population Plant height distribution at Research Park, Ardmore, Okla Parental distribution Agrfa103-2 R June June August December Both the parents grew equally in June December Summer dormant parent didn t grow in August but actively grew in December Plant height (cm) Population distribution

20 QTL for plant height on Agrfa103-2 map August 2011 (no growth on dormant genotypes) Linkage groups

21 Crossing program to develop a NAM population of 4,000 genotypes Recurrent Parents Agrfa Parents F 1 generation x x x x x x x x x x Chain cross 10 hybrid progeny for each of the 10 families F 2 generation individual genotypes x 10 F 1 parents = 200 individuals per F 2 population 200 individuals x 20 F 2 populations = 4000 individuals in the NAM population

22 Summer-dormant breeding population evaluations Summer-dormant breeding populations have been developed at the Foundation Populations evaluated at multiple locations in TX and OK NFTF1800 had the best stand at Burneyville, OK NFTF1700 ranked top at Vernon, TX Across locations, NFTF1700 ranked #1 Plant stand of Flecha was statistically identical NFTF1800 NFTF1700 Flecha MaxQ

23 Summary Summer-dormant tall fescue has great potential in the southern Great Plains Long day is critical for inducing summer dormancy Morphotype specific molecular markers have developed Identification of QTLs associated with summer dormancy is in progress Summer dormant breeding populations developed at the Foundation showed their promise in the region

24 Collaborators within Foundation Forage improvement Division Joe Bouton Twain Butler Carolyn Young Plant Biology Division Patrick Zhao Yuhong Tang Greenhouse staff Forage analysis lab Microarray core facility Collaborators outside Foundation AgResearch, New Zealand Gentos, Argentina Texas A&M University University of Missouri DArT Technology, Australia Mark Sorrells, Cornell University

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