Comparative transcriptome analysis provides insights into the resistance regulation mechanism and inhibitory effect of fungicide phenamacril in Fusarium asiaticum.

Fusarium asiaticum is a destructive phytopathogenic fungus that causes Fusarium head blight of wheat (FHB), leading to serious yield and economic losses to cereal crops worldwide. Our previous studies indicated that target-site mutations (K216R/E, S217P/L, or E420K/G/D) of Type I myosin FaMyo5 conferred high resistance to phenamacril. Here, we first constructed one sensitive strain H1S and three point mutation resistant strains HA, HC and H1R. Then we conducted comparative transcriptome analysis of these F. asiaticum strains after 1 and 10 µg·mL-1 phenamacril treatment. Results indicated that 2135 genes were differentially expressed (DEGs) among the sensitive and resistant strains. The DEGs encoding ammonium transporter MEP1/MEP2, nitrate reductase, copper amine oxidase 1, 4-aminobutyrate aminotransferase, amino-acid permease inda1, succinate-semialdehyde dehydrogenase, 2, 3-dihydroxybenzoic acid decarboxylase, etc., were significantly up-regulated in all the phenamacril-resistant strains. Compared to the control group, a total of 1778 and 2097 DEGs were identified in these strains after 1 and 10 µg·mL-1 phenamacril treatment, respectively. These DEGs involved in 4-aminobutyrate aminotransferase, chitin synthase 1, multiprotein-bridging factor 1, transcriptional regulatory protein pro-1, amino-acid permease inda1, ATP-dependent RNA helicase DED1, acetyl-coenzyme A synthetase, sarcoplasmic/endoplasmic reticulum calcium ATPase 2, etc., showed significantly down-regulated expression in phenamacril-sensitive strain but not in resistant strains after phenamacril treatment. In addition, cyanide hydratase, mating-type protein MAT-1, putative purine nucleoside permease, plasma membrane protein yro2, etc., showed significantly co-down-regulated expression in all the strains after phenamacril treatment. Taken together, This study provides deep insights into the resistance regulation mechanism and the inhibitory effect of fungicide phenamacril and these new annotated proteins or enzymes are worth for the discovery of new fungicide targets.

Spatial and temporal regeneration patterns within gaps in the primary forests vs. secondary forests of Northeast China.

Forest gaps play an important role during forest succession in temperate forest ecosystems. However, the differences in spatial distribution and replacement patterns of woody plants (trees and shrubs) between primary and secondary forests remain unclear during the gap-filling processes, especially for temperate forests in Northeast China. We recorded 45,619 regenerated trees and shrubs in young gaps (<10 years), old gaps (10~20 years), and closed forest stands (i.e., filled gaps) in the primary broadleaved Korean pine (Pinus koraiensis Sieb. Rt Zucc.) forests vs. secondary forests (degraded from primary forests). The gap-filling processes along horizontal (Cartesian coordinate system) and vertical (lower layer: 0~5 m, medium layer: 5~10 m, and upper layer: >10 m) dimensions were quantified by shade tolerance groups of trees and shrubs. We found that gap age, competition between species, and pre-existing regeneration status resulted in different species replacement patterns within gaps in primary vs. secondary forests. Gap formation in both primary and secondary forests increased species richness, with 33, 38, 39, and 41 in the primary closed stands, primary forest gaps, secondary closed stands, and secondary forest gaps, respectively. However, only 35.9% of species in primary forest gaps and 34.1% in secondary forest gaps successfully reached the upper layer. Based on the importance values (IVs) of tree species across different canopy heights, light-demanding trees in the upper layer of the secondary forests were gradually replaced by intermediate and shade-tolerant trees. In the primary forests, Korean pine exhibited intermittent growth patterns at different canopy heights, while it had continuous regeneration along vertical height gradients in the secondary forests. The differences in Korean pine regeneration between the primary and secondary forests existed before gap formation and continued during the gap-filling processes. The interspecific competition among different tree species gradually decreased with increasing vertical height, and compared to the primary forests, the secondary forests showed an earlier occurrence of competition exclusion within gaps. Our findings revealed the species replacement patterns within gaps and provided a further understanding of the competition dynamics among tree species during the gap-filling processes.

Comparative transcriptome analysis reveals the resistance regulation mechanism and fungicidal activity of the fungicide phenamacril in Fusarium oxysporum.

Fusarium oxysporum (Fo) is an important species complex of soil-borne pathogenic fungi that cause vascular wilt diseases of agricultural crops and some opportunistic diseases of humans. The fungicide phenamacril has been extensively reported to have antifungal activity against Fusarium graminearum and Fusarium fujikuroi. In this study, we found that the amino acid substitutions (V151A and S418T) in Type I myosin FoMyo5 cause natural low resistance to phenamacril in the plant pathogenic Fo isolates. Therefore, we compared the transcriptomes of two phenamacril-resistant Fo isolates FoII5, Fo1st and one phenamacril-sensitive isolate Fo3_a after 1 µg/mL phenamacril treatment. Among the 2728 differentially expressed genes (DEGs), 14 DEGs involved in oxidation-reduction processes and MFS transporters, were significantly up-regulated in phenamacril-resistant isolates. On the other hand, 14 DEGs involved in ATP-dependent RNA helicase and ribosomal biogenesis related proteins, showed significantly down-regulated expression in both phenamacril-resistant and -sensitive isolates. These results indicated that phenamacril not only seriously affected the cytoskeletal protein binding and ATPase activity of sensitive isolate, but also suppressed ribosome biogenesis in all the isolates. Hence, this study helps us better understand resistance regulation mechanism and fungicidal activity of phenamacril and provide reference for the development of new fungicides to control Fo.