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Botany JoVE (Open Access) Citable · DOI

Establishing Fungal Entomopathogens as Endophytes: Towards Endophytic Biological Control

DOI: 10.3791/50360-v
What you'll learn
  • ✓ Perform two fungal inoculation methods for endophyte establishment in bean plants
  • ✓ Culture and prepare Beauveria bassiana for plant inoculation
  • ✓ Evaluate fungal colonization and endophytic establishment in plant tissues
Protocol

This protocol demonstrates two inoculation methods to introduce the fungal entomopathogen Beauveria bassiana as an endophyte in the common bean (Phaseolus vulgaris), in preparation for subsequent evaluations of endophytic biological control.

Difficulty
intermediate
Total time
~2–3 weeks (including fungal culturing, inoculation, and colonization assessment)
Model organism
Phaseolus vulgaris (common bean)
Biosafety
BSL-1

Steps

1
Review fungal endophyte and biocontrol background

Understand the rationale for using Beauveria bassiana as an endophyte for pest management and the theoretical basis for endophytic biological control in plants.

▶ 00:04
2
Prepare and acclimate bean plants

Grow Phaseolus vulgaris seedlings under controlled conditions to appropriate developmental stage for inoculation.

▶ 02:30
3
Culture and prepare fungal inoculum

Grow Beauveria bassiana cultures on suitable media and prepare spore or mycelial suspensions for plant inoculation.

▶ 03:51
4
Apply fungal inoculation methods

Introduce B. bassiana as endophyte using two distinct inoculation techniques (e.g., root dip, foliar spray, or direct injection) on bean plants.

▶ 08:22
5
Assess fungal colonization and endophytic establishment

Sample plant tissues and use isolation or molecular methods to confirm B. bassiana presence and distribution within plant tissues over time.

▶ 09:31
6
Summarize establishment results and outcomes

Present data on fungal recovery, colonization patterns, and viability of endophytic B. bassiana for subsequent biocontrol experiments.

▶ 12:43
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