Scientia Agricultura Sinica ›› 2021, Vol. 54 ›› Issue (13): 2759-2768.doi: 10.3864/j.issn.0578-1752.2021.13.006

• PLANT PROTECTION • Previous Articles     Next Articles

The Actin Binding Protein FgAbp1 is Involved in Growth, Development and Toxisome Formation in Fusarium graminearum

ZHANG ChengQi(),WANG XiaoYan,CHEN Li()   

  1. School of Plant Protection, Anhui Agricultural University/Anhui Province Key Laboratory of Integrated Pest Management on Crops/Key Laboratory of Biology and Sustainable Management of Plant Diseases and Pests of Anhui Higher Education Institutes, Hefei 230036
  • Received:2020-10-11 Revised:2020-10-28 Online:2021-07-01 Published:2021-07-12
  • Contact: Li CHEN E-mail:zhcq@ahau.edu.cn;chenlii@ahau.edu.cn

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Abstract:

【Objective】Abp1 is one of the actin binding proteins that plays a central role in actin cytoskeleton of diverse eukaryotic organisms. The objective of this study is to analyze functions of the actin binding protein FgAbp1 in growth and development, sensitivity to the novel fungicide phenamacril and toxisome formation inFusarium graminearum.【Method】Targeted gene deletion construct and fluorescent protein fusion vectors were generated by double-joint PCR and budding yeast gap repair system, respectively. Then, the mutant ΔFgAbp1 and fluorescently labeled strains were obtained using polyethylene glycol (PEG) mediated protoplast transformation. Mycelia growth, sexual/asexual reproduction and sensitivity to phenamacril of wild type PH-1, the mutant ΔFgAbp1 and complemented strain ΔFgAbp1-C were investigated. Localization of FgAbp1 in hyphae was examined through fusion green fluorescent protein. Transmission electron microscopy was carried out to assay the role of FgAbp1 in vacuole/vesicle morphology. Under noninducing medium and DON biosynthesis induction conditions, the role of FgAbp1 in the toxisome formation of F. graminearum was performed by dual fluorescence colocalization assay.【Result】FgAbp1 is primarily localized near the cell membrane in patches of F. graminearum. In MM medium, the growth rate of gene knockout mutant ΔFgAbp1 was reduced by 15% compared with the wild type. But in the nutrient-rich CM, the growth rate of ΔFgAbp1 was decreased by 38%. The mutant ΔFgAbp1 had no obvious defects in sexual and asexual reproduction in comparison with the wild type, while the mycelial growth of ΔFgAbp1 was completely inhibited and the conidia showed significant reduction of germination rate with 0.5 μg·mL -1 phenamacril treatment. Moreover,FgAbp1 deletion resulted in a high vesicle number and a block of normal vacuole formation. During growth in a toxin noninducing condition, FgAbp1 and the DON biosynthetic key enzyme Tri1 co-fluoresced in vesicles. Unexpectedly, FgAbp1 and Tri1 cellular co-localized in toxisomes under DON biosynthesis inducing conditions. Furthermore, disruption of FgAbp1resulted in abnormal toxisomes.【Conclusion】The actin binding protein FgAbp1 plays an important role in vegetative growth, development, phenamacril tolerance and toxisome formation inF. graminearum.

Key words: Fusarium graminearum, FgAbp1, phenamacril, toxisome

Table 1

Primers used in this study"

引物 Primer 序列 Sequence (5′-3′)
Abp1-up-F CAGTGTCGGGGTATTCGGAAGCG
Abp1-up-R CAAAATAGGCATTGATGTGTTGACCTCCGATTGAGGGACGCCATCTTGAGG
Abp1-down-F CTCGTCCGAGGGCAAAGGAATAGAGTAGGGAGTTCCCTGATGAGGATTGG
Abp1-down-R CAACAACCTCAACCTGTAGGCCA
HPH-F GGAGGTCAACACATCAATGCCTATT
HPH-R CTACTCTATTCCTTTGCCCT
Abp1-nest-F GCGTGGAACGATACGATGACGAG
Abp1-nest-R GCTAACCGTTCTATTTAACAGTCC
Abp1-ID-F GACCTTCGTCAGCTTCGTCTCTC
Abp1-ID-R GCCGGGAACAGACCAGTATGGC
Abp1-GFP-F ACTCACTATAGGGCGAATTGGGTACTCAAATTGGTTTTGGAAGCGCCAAATTCTACT
Abp1-GFP-R CACCACCCCGGTGAACAGCTCCTCGCCCTTGCTCACCTGGTTAAGCTGAACGTAGTT
ID-Abp1-GFP-F GCCGCCGCTCAGGCAGATGCC
ID-GFP-R GTCAGCTTGCCGTAGGTGGCA
Abp1-RFP-F ACTCACTATAGGGCGAATTGGGTACTCAAATTGGTTTTGGAAGCGCCAAATTCTACT
Abp1-RFP-R CATGAACTCCTTGATGACGTCCTCGGAGGAGGCCATCTGGTTAAGCTGAACGTAGTT
ID-RFP-R TTGGAGCCGTACTGGAACTG
Tri1-GFP-F ACTCACTATAGGGCGAATTGGGTACTCAAATTGGTTGTGAGTAGGCCTCATAGCGAC
Tri1-GFP-R CACCACCCCGGTGAACAGCTCCTCGCCCTTGCTCACGTCATCCTGTACCAATTCCAAT

Fig. 1

Effects of FgAbp1 deletion on mycelial growth of F. graminearum "

Fig. 2

Asexual and sexual development of gene disruption mutant ΔFgAbp1"

Fig. 3

Sensitivity of mutant ΔFgAbp1 to the fungicide phenamacril"

Fig. 4

Blocking of vacuole formation in the mutant ΔFgAbp1"

Fig. 5

FgAbp1 regulates toxisome formation in F. graminearum "

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