Optimization of the TRV-based Gene Silencing Efficiency and Growth Condition on Solanum lycopersicum var. ‘Hawaii7996’ Resistant to Bacterial Wilt Disease

Nayoung Kim; Won-Hee Kang; Ji-Su Kwon; Hyo-Jin Kim; Boseul Park; Seon-In Yeom

doi:10.7235/HORT.20230039

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2023 Vol.41, Issue 4 Preview Page Next Page

Research Article

Optimization of the TRV-based Gene Silencing Efficiency and Growth Condition on Solanum lycopersicum var. ‘Hawaii7996’ Resistant to Bacterial Wilt Disease 풋마름병 저항성 토마토의 TRV 기반 유전자 침묵 효율 검증 및 생육 조건 최적화

31 August 2023. pp. 429-436

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Abstract

Ralstonia solanacearum (R. sol), a pathogen that causes wilt symptoms, is widely distributed in many regions. Breeding plants resistant against this pathogen is well known as the most effective control method. To this end, it is necessary to reveal the function of resistance genes, and virus- induced gene-silencing (VIGS) technology is one of the most widely used methods for analyzing gene functions. However, the problem with functional analysis of bacterial wilt resistance genes based on VIGS is the conflicting temperature environments of inoculated strains. Therefore, the objective of this study is to identify optimal plant size and temperature conditions for experiments using two srains considering a less complicated functional analysis of R. sol resistant candidate genes. We applied five different temperatures and then assessed the disease progress of R. sol inoculated tomatoes. The plants with the highest VIGS efficiency were those treated at 22°C for four weeks (4L), three weeks at a low temperature and one week at 30°C (3L1H), and two weeks at 22°C and two weeks at 30°C (2L2H). Also, we confirmed the most suitable temperature for identifying candidate genes against R. sol was found to be 22°C for plants grown for three weeks, followed by 30°C for one week. The results of this study can provide useful data to those seeking to establish a foundation for functional analysis research focusing on genes related to tomato bacterial wilt resistance.

Keywords

optimal temperature

Ralstonia solanacearum

tomato

VIGS (Virus-induced gene silencing)

Ralstonia solanacearum는 식물의 물관을 막아 시들음 증상을 유발하며 다양한 지역에 광범위하게 분포되어있어 전세계 토마토 생산량에 심각한 영향을 주는 병원균이다. 이를 해결하기 위해 알려진 가장 효율적인 방법은 저항성 품종 육성으로 다양한 방법 중 하나인 바이러스 기반 유전자 침묵(Virus-Induced Gene Silencing, VIGS)기술을 이용하여 풋마름병에 대한 저항성 후보유전자의 발현을 억제 혹은 감소시켜 기능을 분석할 수 있다. VIGS를 기반으로 한 풋마름병 저항성 유전자 기능 분석 과정에서는 접종 균주의 온도 환경 차이가 상충된다는 문제점이 있다. 따라서 VIGS 적용 후 풋마름병원균 접종을 위한 최적 온도 조건과 이에 따른 식물 생장 조건을 검증하였다. 이를 위해 5가지 생육조건에 따른 VIGS 효율을 확인하고 이후 풋마름병원균 접종을 통해 토마토의 병 진전을 평가하였다. 그 결과, VIGS 효율이 높은 식물체는 4주간 저온(4L), 3주 저온 후 고온 1주(3L1H), 그리고 2주 저온 후 2주 고온(2L2H)을 처리한 식물체였으며 풋마름병을 접종하였을 때 저항성 대조구인 ‘Hawaii 7996’과 유사한 양상을 보이는 3주간 저온 후 1주간 고온 처리한 3L1H 개체가 풋마름병 저항성 후보유전자 발굴에 가장 효과적인 식물체임을 확인하였다. 이 연구 결과는 향후 토마토 풋마름병 저항성 관련 유전자의 기능 분석 연구 기반 구축에 유용한 데이터로 사용될 것이다.

키워드

온도 최적화

Ralstonia solanacearum

토마토

VIGS(Virus-induced gene silencing)

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Information

Publisher :KOREAN SOCIETY FOR HORTICULTURAL SCIENCE
Publisher(Ko) :한국원예학회
Journal Title :Horticultural Science and Technology
Journal Title(Ko) :원예과학기술지
Volume : 41
No :4
Pages :429-436
Received Date : 2022-07-13
Revised Date : 2022-12-15
Accepted Date : 2022-12-22
DOI :https://doi.org/10.7235/HORT.20230039

Horticultural Science and Technology 원예과학기술지 ISSN:1226-8763(Print) 2465-8588(Online)

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