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Peroxidase Gene Polymorphism in Buffalograss and Other Grasses

O. Gulsen, Alata Horticultural Research Institute, Mersin 33740, Turkey; R.C. Shearman and D.J. Lee, Dep. of Agronomy and Horticulture, Univ. of Nebraska, Lincoln, NE 68583; T.M. Heng-Moss, Dep. of Entomology, Univ. of Nebraska, Lincoln, NE 68583; N. Mutlu, West Mediterranean Agricultural Research Institute, Antalya 07110, Turkey; G. Sarath, USDA-ARS, Lincoln, NE 68583

View larger version (48K): [in this window] [in a new window]   Figure 1. Unweighted pair group method arithmetic average (UPGMA) dendogram with similarity coefficients for 28 buffalograsses based on analysis of 52 markers as amplified with peroxidase primers is presented here. Chinch bug resistance is identified in the second column as highly resistant (HR), moderately resistant (MR), moderately susceptible (MS), and highly susceptible (HS) (Gulsen et al., 2004).

View larger version (18K): [in this window] [in a new window]   Figure 2. Genomic DNA was amplified by primer pair POX2 and separated at 90 V for 4 to 5 h. Agarose gel (2.5%) images of 28 buffalograss genotypes are as follows: (1) 184, (2) Prestige, (3) 196, (4) PX3-5-1, (5) 240, (6) 193, (7) 209, (8) 170, (9) 83, (10) 203, (11) 47, (12) Density, (13) 174, (14) 45B, (15) 132, (16) 97, (17) 95-55, (18) 87A, (19) 28, (20) 378, (21) 223A, (22) III-4-9, (23) II-6-6, (24) III-6-6, (25) DP-47-G, (26) 4A, (27) 188, and (28) 119; S = standard marker (DNA Ladder VI; Roche Corp., Indianapolis, IN).

View larger version (38K): [in this window] [in a new window]   Figure 3. Agarose gel (2.5%) images of five C4 grasses (buffalograss, Buchloe dactyloides; sorghum, Sorghum bicolor; zoysiagrass, Zoysia japonica; switchgrass, Panicum virgatum; and bermudagrass, Cynodon dactylon) and four C3 grasses (barley, Hordeum vulgare; rice, Oryza sativa; wheat, Triticum aestivum; rye, Secale cereale) are presented. Genomic DNA was amplified by primer pairs POX2 and POX8 separated at 90 V for 4 to 5 h as follows: (1) buffalograss, (2) barley, (3) rice, (4) wheat, (5) sorghum, (6) zoysiagrass, (7) bermudagrass, (8) switchgrass, and (9) rye. S = standard marker (DNA Ladder VI, Roche Corp., Indianapolis, IN).

View larger version (8K): [in this window] [in a new window]   Figure 4. Unweighted pair group method arithmetic average (UPGMA) dendogram of five C4 grasses (buffalograss, Buchloe dactyloides; bermudagrass, Cynodon dactylon; switchgrass, Panicum virgatum; sorghum, Sorghum bicolor; and zoysiagrass, Zoysia japonica) and four C3 grasses (barley, Hordeum vulgare; rice, Oryza sativa; rye, Secale cereale; and wheat, Triticum aestivum) based on analysis of 64 markers as amplified with peroxidase primers.

View larger version (12K): [in this window] [in a new window]   Figure 5. Phylogenetic relationships among the peroxidase genes sequenced in this study were constructed with Vector NTI Suite Software version 8.0 (Invitrogen Corp., Carlsbad, CA), using CLUSTAL W module. The upper branch includes buffalograss peroxidase genes, while the lower branch includes peroxidase genes from the other grasses studied.

View larger version (27K): [in this window] [in a new window]   Figure 6. Phylogenetic relationships between the rice peroxidase genes used to design primers and peroxidase genes sequenced in this study. The tree was constructed with CLUSTAL W algorithm nested in Vector NTI Suite Software Version 8.0 (Invitrogen Corp., Carlsbad, CA). The rice peroxidase cDNA sequences were obtained from the National Center for Biotechnology Information website (http://www.ncbi.nlm.nih.gov/).