Comparative Study of the Co-Occurring Alternaria and Colletotrichum Species in the Production of Citrus Leaf Spot

Both of the two citrus diseases, Alternaria brown spot (ABS) and Anthracnose, caused by Alternaria and Colletotrichum spp., respectively, can produce leaf lesions which are hard to differentiate. These two diseases have been confused as causal agents of brown spot for over a decade in China. In this study, citrus leaves with or without brown spot were collected from Zhaoqing, Guangdong and Wanzhou, Chongqing, and were further used for the taxonomic and functional comparisons between the co-occurring Alternaria and Colletotrichum species. In the amplicon sequencing, the average relative abundance and the composition of Alternaria, but not Colletotrichum, increased (from 0.1 to 9.9, p = 0.059; and to 0.7, p < 0.05) and significantly altered (p < 0.01) with the brown spot in Zhaoqing and Wanzhou, respectively. Two representative isolates Alternaria sp. F12A and Colletotrichum sp. F12C, from the same brown spot, were proved with different virulence and host response activation to citrus leaves. F12A caused typical symptoms of brown spot with the average spot length expanded to 5 and 6.1 cm, and also altered the citrus global gene expression 48 and 72 h after inoculation. In addition, F12A enriched the expression of genes that were most frequently involved in plant defense. In comparison, F12C caused leaf spot limited to the wounded site, and its milder activation of host response recovered 72 h after inoculation. Our study indicates that the incidence of brown spot in China is caused by Alternaria species, and the ABS should be a fungal disease of major concern on citrus.