To evaluate the copy number of CtDXR gene in transgenic Nicotiana benthamiana and further analyze its tolerance to salt and heat stress,Southern Blot was applied to determine the copy number of the Actin gene in wild-type N.benthamiana and the copy number of the CtDXR gene in CtDXR transgenic N.benthamiana was detected by real-time PCR with Actin gene as reference gene.Then,the salt and heat tolerance of CtDXR transgenic N.benthamiana was analyzed.According to the PCR-dependent method,10 of T1 transgenic N.benthamiana were randomly detected and all of which obtained positive results,indicating that CtDXR has been suc-cessfully transformed.The Actin gene is a single-copy gene in the genome of wild-type N.benthamiana.80%of transgenic N.benthamiana were determined as single-copy or low-copy lines.CtDXR transgenic N.benthamiana showed better performance,such as plant height(P<0.01),number of lateral roots(P<0.01),and fresh weight(P<0.001),than wild-type N.benthamiana under salt stress,whereas CtDXR transgenic N.benthamiana exhibited better performance,such as plant height(P<0.01),number of leaves(P<0.01),and fresh weight,than wild-type N.benthamiana under heat stress.Taken together,CtDXR conferred salt and heat tolerance to N.benthamiana.In this study,a method to detect copy number of foreign gene in transgenic N.benthamiana was established,by which the copy number of CtDXR gene in transgenic N.benthamiana was accomplished.Moreover,CtDXR gene was preliminarily identified to have salt and heat tolerance.
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