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An effective and low-cost culture medium for isolation and growth of Xylella fastidiosa from citrus and coffee

S A Lopes1, S C Z Torres

  • 1Fundo de Defesa da Citricultura, Caixa Postal 391, 14807-040, Araraquara, SP. slopes@fundecitrus.com.br

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Area of Science:

  • Plant Pathology
  • Microbiology
  • Agricultural Science

Background:

  • Buffered charcoal-yeast extract medium (BCYE) is standard for isolating Xylella fastidiosa.
  • BCYE's components, like ACES buffer, L-cysteine, and ferric pyrophosphate, present cost and sterilization challenges.
  • Xylella fastidiosa causes significant diseases in citrus (citrus variegated chlorosis) and coffee (coffee leaf scorch).

Purpose of the Study:

  • To develop a more affordable and accessible medium for Xylella fastidiosa isolation.
  • To create a fully autoclavable medium, simplifying preparation.
  • To evaluate the efficacy of the new medium compared to BCYE.

Main Methods:

  • Modified BCYE by omitting L-cysteine, adding magnesium sulfate, and replacing ACES buffer and ferric pyrophosphate with potassium phosphate and ferrous sulfate.
  • Developed a new medium named phosphate buffered charcoal-yeast extract medium (PCYE).
  • Tested PCYE for Xylella fastidiosa isolation from symptomatic citrus and coffee leaves.

Main Results:

  • PCYE is approximately one-tenth the cost of BCYE.
  • PCYE demonstrated equal effectiveness to BCYE in isolating Xylella fastidiosa from infected plant tissues.
  • The new medium is entirely autoclavable, eliminating the need for filter sterilization of certain components.

Conclusions:

  • Phosphate buffered charcoal-yeast extract medium (PCYE) is a viable, cost-effective alternative to BCYE for Xylella fastidiosa isolation.
  • PCYE simplifies laboratory procedures due to its autoclavable nature.
  • This development supports routine isolation, growth, and quantification of Xylella fastidiosa in research and diagnostics.