Botrytis 속 병원균의 담배 접종체계 확립 및 병저항성 관련 AOS 및 LOX 유전자 발현량 검정
Establishment of an inoculation system for pathogens of the Botrytis genus in Nicotiana tabacum L. cv Havana SR1 and validation of AOS and LOX gene expression levels related to disease resistance
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The Botrytis genus comprises necrotrophic pathogens that damage various horticultural crops worldwide. Nicotiana tabacum is used as a model plant because it can produce the next generation within three months and generate millions of seeds from one plant. In the case of crops that have features difficult to transform, N. tabacum can be used for transformation to validate unknown genes. The objective of this study was to establish inoculation systems of N. tabacum cv Havana SR1 against two pathogens of the Botrytis genus. As a result, it was appropriate to inoculate 10 µL of conidia suspension (2.25 × 105 cells/µL) using unwounded, in vitro cultured leaves, followed by cultivation at 24°C. In addition, to confirm the resistance response to pathogens at the genetic level, the expression of lipoxygenase (LOX) and allene oxide synthase (AOS), which are jasmonic acid (JA) biosynthetic genes involved in signaling when infected with necrotrophic pathogens, was analyzed. The results showed that NtLOX and NtAOS genes were responsive Botrytis inoculation, with peak expression levels observed at 4 hours post-inoculation. It was also confirmed that LOX and AOS genes are closely related to disease resistance in Botrytis genus. The inoculation system established in this study using N. tabacum is expected to contribute to identifying the function of genes related to resistance to B. cinerea or B. squamosa in crops that are difficult to transform
Abstract
The Botrytis genus comprises necrotrophic pathogens that damage various horticultural crops worldwide. Nicotiana tabacum is used as a model plant because it can produce the next generation within three months and generate millions of seeds from one plant. In the case of crops that have features difficult to transform, N. tabacum can be used for transformation to validate unknown genes. The objective of this study was to establish inoculation systems of N. tabacum cv Havana SR1 against two pathogens of the Botrytis genus. As a result, it was appropriate to inoculate 10 µL of conidia suspension (2.25 × 105 cells/µL) using unwounded, in vitro cultured leaves, followed by cultivation at 24°C. In addition, to confirm the resistance response to pathogens at the genetic level, the expression of lipoxygenase (LOX) and allene oxide synthase (AOS), which are jasmonic acid (JA) biosynthetic genes involved in signaling when infected with necrotrophic pathogens, was analyzed. The results showed that NtLOX and NtAOS genes were responsive Botrytis inoculation, with peak expression levels observed at 4 hours post-inoculation. It was also confirmed that LOX and AOS genes are closely related to disease resistance in Botrytis genus. The inoculation system established in this study using N. tabacum is expected to contribute to identifying the function of genes related to resistance to B. cinerea or B. squamosa in crops that are difficult to transform
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