4.4 Article

Isolation of genes related to abscisic acid production in Botrytis cinerea TB-3-H8 by cDNA-AFLP

Journal

JOURNAL OF BASIC MICROBIOLOGY
Volume 54, Issue 3, Pages 204-214

Publisher

WILEY
DOI: 10.1002/jobm.201200311

Keywords

Botrytis cinerea; Abscisic acid (ABA); cDNA-amplified fragment length polymorphism (cDNA-AFLP); Real-time reverse transcription polymerase chain reaction (real-time PCR)

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Funding

  1. Knowledge Innovation Program of the Chinese Academy of Sciences [KSCX2-EW-J-22, KSCX2-EW-G-16-01]
  2. Frontier Program of Chengdu Institute of Biology, the Chinese Academy of Sciences
  3. West Light Foundation of the Chinese Academy of Science [Y1C5031]

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Abscisic acid (ABA) plays important roles in many aspects of plant growth and development. Botrytis cinerea TB-3-H8, a high-yield strain of ABA, was used to elucidate the molecular mechanisms of ABA production in the present work. cDNA-amplified fragment length polymorphism (cDNA-AFLP) technique was applied to isolate genes differentially expressed between ABA high and low-yield conditions. This resulted in the identification of 856 differentially expressed transcript-derived fragments (TDFs). Forty-five TDFs that displayed obvious up-regulated expression profiles in the ABA high-yield condition were sequenced. Based on BlastX in NCBI, 31 TDFs were assumed to have homology with genes encoding proteins with known functions. According to molecular function of gene ontology (GO) analysis, the 31 TDFs were categorized to proteins with enzyme catalytic activities, transcription factor activities, transporter activities, and kinds of binding activities. Further confirmation of the differential expression of these sequences was carried out by performing semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) on 10 randomly selected TDFs. Five up-regulated genes were selected to analyze the expression profiles using real-time PCR. This study enriches our knowledge of the molecular basis for ABA biosynthesis in B. cinerea TB-3-H8.

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