Detection and Quantification of Anthracnose Pathogen Colletotrichum fructicola in Cultivated Tea-Oil Camellia Species from Southern China Using a DNA-Based qPCR Assay.

Tea-oil Camellia species as edible-oil producing trees are widely cultivated in southern China. Camellia anthracnose that is mainly caused by Colletotrichum fructicola is a major disease of tea-oil trees. However, rapid detection and precise quantification of C. fructicola in different Camellia species that are crucial for the fundamental study of this pathosystem and effective disease management remain largely unexplored. Here, we developed a sensitive, rapid, and accurate method for quantifying C. fructicola growth in different Camellia species using a quantitative PCR assay. Amplified C. fructicola DNA using ITS-specific primers is relatively compared with the amplification of Camellia oleifera using the TUB gene. We determined that the fungal growth is tightly associated with the disease development in Ca. oleifera following C. fructicola infection in a time-course manner. This assay is highly sensitive, as fungal growth was detected in six different inoculated tea-oil Camellia species without visible disease lesion symptoms. Additionally, this method was validated by quantifying the Camellia anthracnose in orchards that did not show any disease symptoms. This assay enables the rapid, highly sensitive, and precise detection and quantification of C. fructicola growth in different tea-oil Camellia species, which will have a practical application for early diagnosis of anthracnose disease under asymptomatic conditions in Camellia breeding and field and will facilitate the development of tea-oil trees and C. fructicola interaction as a mold system to study woody plant and fungal pathogens interaction.

Four new species of Diaporthe (Diaporthaceae, Diaporthales) from forest plants in China.

Species of Diaporthe inhabit a wide range of plant hosts as plant pathogens, endophytes and saprobes. During trips to collect forest pathogens in Beijing, Jiangxi, Shaanxi and Zhejiang Provinces in China, 16 isolates of Diaporthe were obtained from branch cankers and leaf spots. These isolates were studied by applying a polyphasic approach including morphological, cultural data, and phylogenetic analyses of the nuclear ribosomal internal transcribed spacer (ITS), calmodulin (cal), histone H3 (his3), partial translation elongation factor-1α (tef-1α) and ß-tubulin (tub2) loci. Results revealed four new taxa, D.celticola, D.meliae, D.quercicola, D.rhodomyrti spp. nov. and two known species, D.eres and D.multiguttulata.

The AVR2-SIX5 gene pair is required to activate I-2-mediated immunity in tomato.

Plant-invading microbes betray their presence to a plant by exposure of antigenic molecules such as small, secreted proteins called 'effectors'. In Fusarium oxysporum f. sp. lycopersici (Fol) we identified a pair of effector gene candidates, AVR2-SIX5, whose expression is controlled by a shared promoter. The pathogenicity of AVR2 and SIX5 Fol knockouts was assessed on susceptible and resistant tomato (Solanum lycopersicum) plants carrying I-2. The I-2 NB-LRR protein confers resistance to Fol races carrying AVR2. Like Avr2, Six5 was found to be required for full virulence on susceptible plants. Unexpectedly, each knockout could breach I-2-mediated disease resistance. So whereas Avr2 is sufficient to induce I-2-mediated cell death, Avr2 and Six5 are both required for resistance. Avr2 and Six5 interact in yeast two-hybrid assays as well as in planta. Six5 and Avr2 accumulate in xylem sap of plants infected with the reciprocal knockouts, showing that lack of I-2 activation is not due to a lack of Avr2 accumulation in the SIX5 mutant. The effector repertoire of a pathogen determines its host specificity and its ability to manipulate plant immunity. Our findings challenge an oversimplified interpretation of the gene-for-gene model by showing requirement of two fungal genes for immunity conferred by one resistance gene.