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Genetic Diversity among Forage Bermudagrass (Cynodon spp.)

Evidence from Chloroplast and Nuclear DNA Fingerprinting

^a Dep. of Field Crops, Akdeniz Univ., Agricultural Faculty, Antalya, 07759 Turkey
^b Dep. of Plant and Soil Sciences, Mississippi State Univ., Mississippi State, MS 39762
^c USDA-ARS, P.O. Box 5367, Mississippi State, MS 39762
^d Dep. of Plant and Soil Science, Alabama A&M Univ., Normal, AL 35762

View larger version (51K): [in a new window]   Fig. 1. AFLP Electropherograms. The x-axis represents the size (bp) of the DNA fragments and the y-axis indicates the amount of the amplified products in arbitrary numbers. Electropherogram A: Red and blue lines indicate AFLP DNA bands of Tifton 78 and Tifton 78 WH, respectively, using the AA-CAA primer pair. Electropherogram B: AFLPs between Callie (red) and Tifton 85 (blue) amplified with the AA-CAA primer pair. Note four polymorphic AFLPs indicated by arrows. Electropherogram C: A typical AFLP electropherogram showing amplified products ranging from 50 to 450 bp in length. The red line is the internal size standard.

View larger version (48K): [in a new window]   Fig. 2. CpSSRLP Electropherograms. An automated capillary electrophoretic system showing polymorphic fluorescently labeled fragments amplified with the CpSSRLP CCMP5R/F primer pair. PCR products are visualized as peaks on electropherograms. The x-axis represents the size (bp) of the DNA fragment and the y-axis shows the amount of the amplified products in arbitrary numbers. The red line is the internal size standard included in each electrophoretic run. Callie (1), paternal parent of Tifton 78, Tifton 78 (2), an F1 hybrid (Tifton 44 x Callie), Tifton 44 (3), a maternal parent of Tifton 78 and Coastal (4), a maternal parent of Tifton 44. Local ecotypes, Murphy (5) and Murphy II (6) showing differences in the chloroplast genome. These two ecotypes were included in the figure to emphasize the power of the CpSSRLP technique for differentiating two local ecotypes.

View larger version (46K): [in a new window]   Fig. 3. DAMD Agarose Gel Electrophoresis. Lanes 1 to 33 are Holly Springs, Tifton 44, Tifton 78, Tifton 78 WH, Callie, Coastal, Lott II, Murphy, Murphy II, Lott I, McDonald, World Feeder, Alicia, ‘Russell’,* Prairie III, Tifton 85, Prairie I, Prairie II, Pasto Rico, Gillihan, Sumrall 007, common type, Stallings, Maddox, Tanberg, Poplarville II, Lancaster, Hardie, Poplarville I, Dixie I, Dixie II, Grazer and negative control amplified using primer, MfVIIe8-C. *Excluded from the analysis due to plant contamination from other sources.

View larger version (42K): [in a new window]   Fig. 4. RAPD Agarose Gel Electrophoresis. Lanes 1 to 33 are Callie (1), Tifton 44 (2), Tifton 78 (3), Tifton 78 WH (4), Coastal (5), Murphy (6), Murphy II (7); Lott I (8), Lott II (9), Prairie (10), Prairie II (11), Prairie III (12), Tifton 85 (13), Pasto Rico (14), World Feeder (15), Alicia (16), Russell* (17), Sumrall 007 (18), common (19), Stallings (20), Maddox (21), Tanberg (22), Poplarville I (23), Poplarville II (24), Lancaster (25), ‘Gillihan’ (26), Holly Springs (27), McDonald (28), Hardie (29), Dixie I (30), Dixie II (31), Grazer (32) bermudagrass lines and negative control (33), respectively, amplified with the RAPD primer OPM-20. *Excluded from the analysis due to plant contamination from other sources.

View larger version (53K): [in a new window]   Fig. 5. Majority-rule consensus UPGMA Trees of 31 Forage Bermudagrass Genotypes. The trees were generated using the distance matrix based on Nei's formula from 15 AFLP, 10 CpSSRLP, 10 RAPD, and 10 DAMD primers or primer pairs. Numbers on branches are bootstrap frequency values for 2000 bootstrap replicates.

View larger version (29K): [in a new window]   Fig. 6. Majority-rule consensus UPGMA tree combining AFLP, CpSSRLP, RAPD and DAMD data. The tree was generated using the distance matrix based on Nei's formula from 15 AFLP, 10 CpSSRLP, 10 RAPD, and 10 DAMD primers or primer pairs. Numbers on branches are bootstrap frequency values for 2000 bootstrap replicates. Note combining information from all four markers systems showed better bootstrap support than any other phylogenies. A, B, C, D, and E are cluster groups discussed in the text.