Curvularia leaf spot (CLS), primarily caused by Curvularia lunata (Wakker) Boedijn (C. lunata), is widely distributed in maize production regions in China. It occurs in all the developmental stages of maize and causes economic losses. The epidemic and yield loss estimation models have been constructed for the disease. C. lunata has obvious virulence differentiation and produces multiple virulence factors. CLS is managed by application of chemical and biological agents and by quantitative resistance conferred by 5 to 6 quantitative trait loci (QTL). This review summarizes research on the understanding of CLS biological characteristics, virulence factors of C. lunata, host resistance genetics, and disease management strategies in China.

Southern corn leaf blight (SCLB) disease caused by Cochliobolus heterostrophus is one of the major threats to corn production worldwide. The synergistic application of low toxic chemical fungicide and biocontrol agents could improve biocontrol stability and efficiency against plant diseases, which ultimately reduce use of chemical fungicide. Trichoderma spp., well-known biocontrol fungi have been used to control some foliar diseases. However, few works have been reported on synergistic application of chemical fungicide and Trichoderma against foliar diseases. This study was aimed to investigate the control effect on the synergistic application of Trichoderma harzianum SH2303 and difenoconazole-propiconazole (DP) against SCLB. Results showed that the synergistic application of DP and SH2303 reduced the leaf spot area compared to the control. The efficacy of synergistic application of DP+SH2303 against SCLB could last for 15–20 d in pot trial under the greenhouse condition. Under the natural field condition, maize treated with DP+DP and DP+SH2303 showed 60% control, which was higher than that of SH2303+DP (45%) and SH2303+SH2303 (35%). All these treatments induced the synthesis of defense-related enzymes (phenylalanine ammonia lyase (PAL), catalase (CAT), and superoxide dismutase (SOD)) and the defence-related gene expression of SA pathway (PR1). Taken together the in-vitro leaf test and field trial, the control of SCLB by synergistic application of DP+SH2303 was similar to that of DP+DP. Among synergistic application, the sequential application of DP+SH2303 showed better control than the sequential application of SH2303+DP. It was concluded that the synergistic application of chemical fungicide (DP) and biocontrol agent (T. harzianum SH2303) could be used to reduce the chemical fungicide and to reduce the SCLB diseases in maize, which provided alternative approach to realize an eco-friendly controlling of the foliar disease.

MicroRNAs (miRNAs) are known to play important roles in the immune response to pathogen infection in different plants. Further, Trichoderma strains are able to activate plant defense responses against pathogen attacks. However, little is known about the involvement of miRNAs in the defense response primed by Trichoderma strains. To explore the miRNAs sensitive to priming by Trichoderma, we studied the small RNAs and transcriptome changes in maize leaves that were systemically induced by seed treatment with Trichoderma harzianum (strain T28) against Cochliobolus heterostrophus (C. heterostrophus) infection in leaves. Through analysis of the sequencing data, 38 differentially expressed miRNAs (DEMs) and 824 differentially expressed genes (DEGs) were identified. GO and KEGG analyses of DEGs demonstrated that genes involved in the plant hormone signal transduction pathway and oxidation-reduction process were significantly enriched. In addition, 15 miRNA–mRNA interaction pairs were identified through the combined analysis of DEMs and DEGs. These pairs were supposed to play roles in the maize resistance primed by T. harzianum T28 to C. heterostrophus, in which miR390, miR169j, miR408b, miR395a/p, and novel miRNA (miRn5231) were more involved in the induction of maize resistance. This study provided valuable information for understanding the regulatory role of miRNA in the T. harzianum primed defense response.

Maize leaf spot, which is caused by Curvularia lunata (Wakkre) Boedijn, was epidemic in the maize-growing regions of northeastern and northern China in the mid-1990s, where it led to large yield losses. Since then, the epidemic has evolved into a kind of common disease. In recent years, however, a tendency of becoming an epidemic disease again has been observed in some areas in China due to significant changes in climate, farming, systems and crop varieties. The significance of methyl-(5hydroxymethyl) furan-2-carboxylate (M5HF2C) as a nonspecific host toxin in causing maize leaf spot disease has been demonstrated in previous research. However, the key enzymes involved in M5HF2C toxin synthesis remain unclear. In our study, we demonstrate that the synthesis of M5HF2C toxin starts from a precursor substrate in the pathogen, furfural, which is then catalytically dehydrogenated into furoic acid via an alcohol dehydrogenase (CLADH6). The furoic acid was further confirmed as one of the raw materials for the biosynthesis of M5HF2C toxin based on deletion mutants of the alcohol dehydrogenase gene (Cladh6) in C. lunata, which had reduced M5HF2C toxin-producing ability; however, this ability could be restored in all deletion mutants through complementation with furoic acid, thereby confirming that furoic acid is an intermediate in the biosynthesis of M5HF2C toxin. In summary, the biosynthesis process of M5HF2C toxin in C. lunata involves three transformation steps: (1) from xylose to furfural; (2) then from furfural to furoic acid; and (3) eventually from furoic acid to M5HF2C toxin. Our research findings provide new clues in elucidating the major steps in the process of M5HF2C toxin biosynthesis in C. lunata.

Cochliobolus heterostrophus is a crucial pathogenic fungus that causes southern corn leaf blight (SCLB) in maize worldwide, however, the virulence mechanism of the dominant race O remains unclear. In this report, the single-cell level of pathogen tissue at three infection stages were collected from the host interaction-situ, and were performed next-generation sequencing from the perspectives of mRNA, circular RNA(circRNA) and long noncoding RNA(lncRNA). In the mRNA section, signal transduction, kinase, oxidoreductase, and hydrolase, et al. were significantly related in both differential expression and co-expression between virulence differential race O strains. The expression pattern of the traditional virulence factors nonribosomal peptide synthetases (NPSs), polyketide synthases (PKSs) and small secreted proteins (SSPs) were multifarious. In the noncoding RNA section, a total of 2279 circRNAs and 169 lncRNAs were acquired. Noncoding RNAs exhibited differential expression at three stages. The high virulence strain DY transcribed 450 more circRNAs than low virulence strain WF. Informatics analysis revealed numbers of circRNAs which positively correlate with race O virulence, and a cross-kingdom interaction between the pathogenic circRNA and host miRNA was predicted. An important exon-intron circRNA Che-cirC2410 combines informatics characteristics above, and highly expressed in the DY strain. Che-cirC2410 initiate from the pseudogene chhtt , which doesn’t translate genetic code into protein. In-situ hybridization tells the sub-cellular localization of Che-cirC2410 include pathogen`s mycelium, periplasm, and the diseased host tissues. The target of Che-cirC2410 was predicted to be zma-miR399e-5P, and the interaction between noncoding RNAs was proved. More, the expression of zma-miR399e-5P exhibited a negative correlation to Che-cirC2410 in vivo. The deficiency of Che-circ2410 decreased the race O virulence. The host resistance to SCLB was weakened when zma-miR399e-5P was silenced. Thus, a novel circRNA-type effector and its resistance related miRNA target are proposed cautiously in this report. These findings enriched the pathogen-host dialogue by using noncoding RNAs as language, and revealed a new perspective for understanding the virulence of race O, which may provide valuable strategy of maize breeding for disease resistance. Author Summary: The southern corn leaf blight (caused by Cochliobolus heterostrophus ) is not optimistic in Asia, however we have limit knowledge about the infection mechanism of the dominant C.heterostrophus race O. We take full advantage of the ideal C.heterostrophus genome database, laser capture microdissection and single-cell level RNA sequencing. Hence, we could avert the artificial influence such as medium, and profile the real gene mobilization strategy in the infection. The results of coding RNA section were accessible, virulence related genes (such as the signal transduction, PKS, SSP) were detected in RNA-seq,which accord with previous reports. However, the results of noncoding RNA was astonished, 2279 circular RNAs (circRNA) and 169 long noncoding RNAs (lncRNA) were revealed in our results. Generally, the function of noncoding RNA was hypothesized in single species, but we boldly guess that the function of circRNA is rather complicated in the pathogen-host interaction. Finally, the circRNA in-situ hybridization (ISH) demonstrate the secretion of pathogen circRNA into the host tissue. By bioinformatic prediction, we found a sole microRNA target, and proved the interaction between circRNA and microRNA. These findings are likely to reveal a novel pathogen effector type: secreted circRNA.