All kinds of crops including foods, feeds, and turf grasses are damaged frequently by various environmental stresses such as drought, salt, and high temperature, which cause the loss of agronomic productivity and the decline in commercial quality. Many abiotic stress responsive genes have been identified in plants and they have been applied to breed crops tolerant to environmental stresses through a transgenic approach. In this study, by use of 4 combinations of degenerate primer PCR and Rsa I, a 4-cutter restriction enzyme recognizing GTAC, 9 independent cDNAs containing NAC domain sequence were isolated from creeping bentgrass (
Figures & Tables
Fig. 1. Isolation of NAC cDNAs from creeping bentgrass using conserved domains of NAC proteins in . (A) Comparison of amino acid sequences of NAC domains consisting of 5 subdomains (A to E) from A. thaliana respectively. (B) Experimental scheme to isolate NAC cDNAs of creeping bentgrass by PCR using degenerate primers based on conserved amino acid sequence domains. (C) Cloning and screening of PCR products into a T-plamid vector. Lane 1 to 4 are RT-PCR products. Lane 1 NAC F1 and NAC R1. Lane 2 NAC F1 and NAC R2. Lane 3 NAC F2 and NAC R1. Lane 4 NAC F2 and NAC R2.