Prediction of Effector Proteins from Trichoderma longibrachiatum Through Transcriptome Sequencing.

Trichoderma longibrachiatum SMF2 is an important biocontrol strain isolated by our group that can promote plant growth and induce plant disease resistance. To further study its biocontrol mechanism, the effector proteins secreted by T. longibrachiatum SMF2 were analyzed through bioinformatics and transcriptome sequencing. Overall, 478 secretory proteins produced by T. longibrachiatum were identified, of which 272 were upregulated after treatment with plants. Functional annotation showed that 36 secretory proteins were homologous with different groups of effectors from pathogenic microorganisms. Moreover, the quantitative PCR results of six putative effector proteins were consistent with those of transcriptome sequencing. Taken together, these findings indicate that the secretory proteins secreted by T. longibrachiatum SMF2 may act as effectors to facilitate its own growth and colonization or to induce plant immunity response.

The gene fpk1, encoding a cAMP-dependent protein kinase catalytic subunit homolog, is required for hyphal growth, spore germination, and plant infection in Fusarium verticillioides.

Fusarium verticillioides is an important pathogen of maize responsible for ear rots, stalk rots and seeding blight worldwide. During the past decade F. verticillioides has caused several severe epidemics of maize seeding blight in many areas of china, which lead to significant losses. In order to understand molecular mechanisms regulating fungal development and pathogenicity in the pathogen, we isolated and characterized the gene fpk1 (GenBank accession NO. EF405959) encoding catalytic subunit of cAMP-dependent protein kinase, which include 1854 bp DNA sequence from ATG to TAA, with 1680 bp coding region, three intron (their length: 66bp, 54bp and 54bp), and the predicated protein had 559 aa. The mutantdeltafpk1, which was disrupted of fpk1 gene, showed reduced vegetative growth, fewer and shorter aerial mycelia, strongly impaired conidiation and reduced spore germination rate. After germinating, the fresh hypha was stubby and lack of branch. When inoculated in susceptible maize varieties, the infection of the mutantdeltafpk1 was delayed and the infection efficiency was reduced than that of the wild-type. All this indicated that the gene fpk1 participated in hyphal growth, conidiophore producing, spore germination and virulence in F. verticillioides.

The FUS3/KSS1-type MAP kinase gene FPK1 is involved in hyphal growth, conidiation and plant infection of Fusarium proliferatum.

Fusarium proliferatum is an important pathogen of maize that is responsible for ear rots, stalk rots and seeding blight worldwide. During the past decade, F. proliferatum has caused several severe epidemics of maize seedling blight in many areas of China, which led to significant losses in maize. To understand the molecular mechanisms in the fungal developmental regulation and pathogenicity, we isolated and characterized the FPK1 gene (GenBank accession No. HQ844224) encoding a MAP kinase homolog of FUS3/KSS1 in yeast. The gene includes a 1,242-bp DNA sequence from ATG to TAA, with a coding region of 1,068 bp, 3 introns (58 bp, 56 bp and 60 bp) and a predicted protein of 355 aa.The mutant ΔFPK1, which has a disruption of the FPK1 gene, showed reduced vegetative growth, fewer and shorter aerial mycelia, strongly impaired conidiation and spore germination, as well as deviant germ tube outgrowth. When the strain was inoculated in susceptible maize varieties, the infection of the mutant ΔFPK1 was delayed, and the infection efficiency was reduced compared to the wild-type strain. Complementation of the disruptions within the FPK1 open reading frame restored wild-type levels of conidiation, growth rate and virulence to maize seedlings. Our results indicated that the FPK1 gene functioned in hyphal growth, conidiation, spore germination and virulence in F. proliferatum.