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A simple protocol for producing axenic seeds of Sorghum bicolor

Summary

Researchers developed a simple method to sterilize sorghum seeds by treating them with ethanol and bleach, which removes contaminating microbes while keeping seeds alive and able to grow. This approach is inexpensive, requires no special equipment, and is useful for scientists studying how microbes interact with plants. The method successfully eliminated harmful microbes from 98% of seeds tested while maintaining a reasonable germination rate of 63%.

Background

Microbes within seeds can confound research on microbial colonization, symbiosis, and pathogenesis. Existing seed sterilization methods for many plant species often fail to eliminate internal culturable microbes. A reliable method that removes culturable microbes from both internal and external seed tissues without compromising germination is needed for plant-microbe interaction studies.

Objective

To develop and validate a simple, cost-effective protocol for producing axenic Sorghum bicolor seeds that reliably eliminates culturable microbes while preserving seed viability for experiments involving microbial inoculation and plant-microbe interactions.

Results

Of 95 seeds tested, only 2 showed contamination from culturable microbes (2.1% contamination rate), both from the same student group. The median germination rate was 63%, with no growth defects observed during subsequent four-week inoculation experiments. The protocol effectively eliminated culturable microbes while maintaining reasonable seed viability.

Conclusion

This protocol provides a reliable, fast, and cost-effective method for producing axenic Sorghum bicolor seeds without requiring specialized equipment. The approach successfully eliminates culturable microbes from both internal and external seed tissues while preserving germination rates suitable for plant-microbe interaction research.
  • Published in:MicroPubl Biol,
  • Study Type:Protocol Development Study,
  • Source: 10.17912/micropub.biology.001772, PMID: 40994815
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