Occurrence of Alternaria leaf blight of sunflower caused by two closely related species Alternaria solani and A. tomatophila in Inner Mongolia

Sunflower (Helianthus annuus L.) is an important oilseed crop in China and is grown in rotation with potato in the northern parts of China, including Inner Mongolia, Hebei, Shaanxi, Shanxi, Gansu, Jilin, Liaoning and Heilongjiang provinces. In 2014, diseased samples of sunflower (variety 3638C) with black sunken lesions on bracts and stems of sunflower were collected in Wuchuan, Hohhot (41°05′ N, 111°27′ E) and Chahaeryouyihouqi, Wulanchabu (41°52′ N, 113°0′ E), Inner Mongolia, China. We used morphological and molecular techniques to identify the causal agent of Alternaria leaf blight. Phylogenetic analysis based on a combined phylogenetic dataset using the glyceraldehyde-3-phosphate dehydrogenase (gpd) and Alternaria major allergen gene (Alt a1) was done to support the morphological identification. In addition, PCR was performed with specific primers targeted on Alt a1 and calmodulin genes of A. solani and A. tomatophila. The results showed that HaAs7 gave an amplicon of the expected size (164 bp) with A. solani-specific primers, however, no band was observed with the A. tomatophila-specific primers. The opposite pattern of PCR result (483 bp) was obtained with DNA of HaAt4 as template, indicating that HaAs7 is A. solani and HaAt4 is A. tomatophila. Furthermore, evaluation the pathogenicity of the recovered Alternaria isolates and their potential as causal agents of Alternaria leaf blight of sunflower was done. Pathogenicity tests showed that A. solani and A. tomatophila tested could be capable of causing Alternaria leaf blight of sunflower. Till now, nine species of Alternaria have been reported causing Alternaria leaf blight of sunflower, this is the first report that A. solani and A. tomatophila as causal pathogens for Alternaria leaf blight on sunflower