MicroRNAs involved in the trans-kingdom gene regulation in the interaction of maize kernels and Fusarium verticillioides.

Maize ear rot is a widespread disease and the main pathogen is Fusarium verticillioides. Plant microRNAs (miRNAs) have great effects on disease resistance and it has been reported that maize miRNA participates in defense responses in maize ear rot. However, the trans-kingdom regulation of miRNAs between maize and F. verticillioides remains uncharacterized. In this study, the relationship between miRNA-like RNAs (milRNAs) of F. verticillioides and pathogenicity was investigated, followed by sRNA analysis and degradome sequencing of miRNA profiles and the target genes of maize and F. verticillioides after inoculation. It was found that the milRNA biogenesis positively regulated the pathogenicity of F. verticillioides by knocking out the gene FvDicer2-encoded Dicer-like protein in F. verticillioides. Following inoculation with F. verticillioides, 284 known and 6571 novel miRNAs were obtained in maize, including 28 miRNAs differentially expressed at multiple time points. The target genes of maize differentially expressed miRNAs in F. verticillioides mediated multiple pathways, including autophagy and MAPK signaling pathway. Fifty-one novel F. verticillioides milRNAs were predicted to target 333 genes in maize involved in MAPK signaling pathways, plant hormone signaling transduction and plant-pathogen interaction pathways. Additionally, the miR528b-5p in maize targeted the mRNA of FvTTP which encoded a twice transmembrane protein in F. verticillioides. The FvTTP-knockout mutants displayed decreased pathogenicity and reduced synthesis of fumonisins. Thus, by interfering with the translation of FvTTP, the miR528b-5p inhibited F. verticillioides infection. These findings suggested a novel function of miR528 in resisting F. verticillioides infection. The miRNAs identified in this research and their putative target genes can be used to further elucidate the trans-kingdom functions of microRNAs in plant pathogen interaction.

Transcriptional differences between major Fusarium pathogens of maize, Fusarium verticillioides and Fusarium graminearum with different optimum growth temperatures.

Fusarium verticillioides and Fusarium graminearum are important pathogens causing disease in maize (Zea mays) worldwide. The distributions of these fungal pathogens vary greatly in different regions and in different years, and are influenced by environmental and climatic conditions. Temperature has significant effects on the growth and mycotoxin production of Fusarium species. In this study, the effects of temperature on the growth and pathogenicity of F. verticillioides and F. graminearum were investigated. F. verticillioides grew fastest and exhibited the strongest pathogenicity to maize stems and grains at 30°C, while F. graminearum grew best at 20°C. Both species produced more toxins at 20°C than at 30°C. To explain the interspecific differences in the relationship of growth and temperature, RNA-seq was used to compare F. verticillioides and F. graminearum cultivated for 4 d at the optimum temperatures of 30°C and 20°C, respectively. Samples of F. verticillioides were also cultivated for 9 d (to maximize toxin production) at 20°C and 30°C and analyzed by RNA-seq to investigate the influence of temperature for different growth stages. The differently expressed genes (DEGs) were identified by comparison of cultures grown for the same amount of time but at different temperatures. GO enrichment analysis showed high enrichment of DEGs in categories of membrane part, catalytic activity, metabolic process, and growth at warmer temperature resulted in more down-regulated DEGs enriched in membrane components in all groups. KEGG analysis revealed enrichment of DEGs related to different temperatures in carbohydrate and amino acid metabolism pathways. For both species, there was decreased expression of many DEGs related to amino acid metabolism when cultivated at warm temperature, such as genes related to beta-alanine metabolism and arginine and proline metabolism. However, changes in genes related to glyoxylate and dicarboxylate metabolism and fatty acid degradation were more related to the growth state. The results showing different responses pattern of these pathways provides a foundation for further investigation of the molecular mechanisms underlying distinct thermal ecological niches of F. verticillioides and F. graminearum.

Maize stalk rot caused by Fusarium graminearum alters soil microbial composition and is directly inhibited by Bacillus siamensis isolated from rhizosphere soil.

Maize stalk rot caused by Fusarium graminearum can reduce the yield of maize and efficiency of mechanized harvesting. Besides, deoxynivalenol and zearalenone toxins produced by F. graminearum can also affect domestic animals and human health. As chemical fungicides are expensive and exert negative effects on the environment, the use of biological control agents has become attractive in recent years. In the present study, we collected rhizosphere soil with severe stalk rot disease (ZDD), the rhizosphere soil with disease-free near by the ZDD (ZDH), and measured rhizosphere microbial diversity and microbial taxonomic composition by amplicon sequencing targeting either bacteria or fungi. The results showed that Fusarium stalk rot caused by the Fusarium species among which F. graminearum is frequent and can reduce the abundance and alpha diversity of rhizosphere microbial community, and shift the beta diversity of microorganisms. Furthermore, a bacterial strain, Bacillus siamensis GL-02, isolated from ZDD, was found to significantly affect growth of F. graminearum. In vitro and in vivo assays demonstrated that B. siamensis GL-02 had good capability to inhibit F. graminearum. These results revealed that B. siamensis GL-02 could be a potential biocontrol agent for the control of maize stalk rot.

Population Source of Third-Generation Oriental Armyworm in Jilin, China, Determined by Entomology Radar, Trajectory Analysis, and Mitochondrial COI Sequences.

The armyworm, Mythimna separata (Walker) (Lepidoptera: Noctuidae), is an important polyphagous pest with a strong migratory ability. Recently, third-generation larvae have become an increasingly serious pest threat in Jilin Province of northeast China. To investigate the population source of this species, scanning entomological radar observations and insect mitochondrial cytochrome oxidase I (COI) genes were used in this study. Five main results were found: (1) The peak period in captured second-generation moths was from mid to late July. The temperature and wind speeds were optimum for the moths to have migrated. Strong southwesterly winds occurred during the peak migration period. (2) Radar observations indicated that most of the moths' migration took place at a height of 600 m, often in a dense layer which formed at heights of 350-800 m. (3) Analyses of adult ovarian development and larval haplotypes showed third-generation larvae were progeny of both locally produced progeny and immigrant moths. (4) Based on our back-tracking and haplotype analyses, immigration led to an outbreak originated in the same source area to the southwest. (5) Emigration of second-generation moths was confirmed by both radar observation and mtDNA analysis. Forward trajectories indicated that the moths were capable of immigrating far from their overwintering range. These results are useful for improving the forecasting systems of this insect pest species.

Genetic variation and geographic differentiation among populations of the nonmigratory agricultural pest Oedaleus infernalis (Orthoptera: Acridoidea) in China.

The nonmigratory grasshopper Oedaleus infernalis Saussure (Orthoptera : Acridoidea) is an agricultural pest to crops and forage grasses over a wide natural geographical distribution in China. The genetic diversity and genetic variation among 10 geographically separated populations of O. infernalis was assessed using polymerase chain reaction-based molecular markers, including the intersimple sequence repeat and mitochondrial cytochrome oxidase sequences. A high level of genetic diversity was detected among these populations from the intersimple sequence repeat (H: 0.2628, I: 0.4129, Hs: 0.2130) and cytochrome oxidase analyses (Hd: 0.653). There was no obvious geographical structure based on an unweighted pair group method analysis and median-joining network. The values of FST, θ(II), and Gst estimated in this study are low, and the gene flow is high (Nm > 4). Analysis of the molecular variance suggested that most of the genetic variation occurs within populations, whereas only a small variation takes place between populations. No significant correlation was found between the genetic distance and geographical distance. Overall, our results suggest that the geographical distance plays an unimpeded role in the gene flow among O. infernalis populations.