Species clarification of Isaria isolates used as biocontrol agents against Diaphorina citri (Hemiptera: Liviidae) in Mexico

Fungal Biol. 2016 Mar;120(3):414-23. doi: 10.1016/j.funbio.2015.11.009. Epub 2015 Dec 17.

Abstract

Entomopathogenic fungi belonging to the genus Isaria (Hypocreales: Cordycipitaceae) are promising candidates for microbial control of insect pests. Currently, the Mexican government is developing a biological control program based on extensive application of Isaria isolates against Diaphorina citri (Hemiptera: Liviidae), a vector of citrus huanglongbing disease. Previous research identified three promising Isaria isolates (CHE-CNRCB 303, 305, and 307; tentatively identified as Isaria fumosorosea) from Mexico. The goal of this work was to obtain a complete morphological and molecular characterization of these isolates. Comparative analysis of morphology established that the isolates showed similar characteristics to Isaria javanica. Multi-gene analysis confirmed the morphological identification by including the three isolates within the I. javanica clade. Additionally, this work demonstrated the misidentifications of three other Isaria isolates (CHE-CNRCB 310 and 324: I. javanica, formerly I. fumosorosea; CHE-CNRCB 393: I. fumosorosea, formerly Isaria farinosa), underlying the need for a full and correct characterization of an isolate before developing a biological control program. Finally, the inter-simple sequence repeat (ISSR) genotyping method revealed that the CHE-CNRCB 303, 305, and 307 isolates belong to three different genotypes. This result indicates that ISSR markers could be used as a tool to monitor their presence in field conditions.

Keywords: Biological control; Entomopathogenic fungi; ISSR markers; Isaria javanica; Morphology; Phylogeny.

MeSH terms

  • Animals
  • Citrus / parasitology
  • Cluster Analysis
  • DNA, Fungal / chemistry
  • DNA, Fungal / genetics
  • DNA, Ribosomal Spacer / chemistry
  • DNA, Ribosomal Spacer / genetics
  • Genotype
  • Hemiptera / microbiology*
  • Hypocreales / classification*
  • Hypocreales / cytology
  • Hypocreales / genetics
  • Hypocreales / isolation & purification
  • Mexico
  • Microscopy
  • Molecular Sequence Data
  • Peptide Elongation Factor 1 / genetics
  • Pest Control, Biological
  • Phenotype
  • Phylogeny
  • Plant Diseases / parasitology
  • Plant Diseases / prevention & control
  • Sequence Analysis, DNA
  • Tubulin / genetics

Substances

  • DNA, Fungal
  • DNA, Ribosomal Spacer
  • Peptide Elongation Factor 1
  • Tubulin