[논문]인삼내생균 Bacillus velezensis CH-15의 인삼뿌리썩음병 방제 효과

김도현; 이정관; 이승호

doi:10.5423/rpd.2022.28.1.19

인삼내생균 Bacillus velezensis CH-15의 인삼뿌리썩음병 방제 효과
Biological Efficacy of Endophytic Bacillus velezensis CH-15 from Ginseng against Ginseng Root Rot Pathogens 원문보기

Research in plant disease = 식물병연구, v.28 no.1, 2022년, pp.19 - 25

김도현 (동아대학교 응용생물공학과) , (동아대학교 응용생물공학과) , 이정관 (동아대학교 응용생물공학과) , 이승호 (국립원예특작과학원 인삼특작부)

초록
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인삼은 수천 년 동안 동아시아에서 재배된 중요한 약용 식물이다. 일반적으로 같은 포장에서 4-6년 동안 재배되기 때문에 다양한 병원균에 노출된다. 그 중 인삼뿌리썩음병이 가장 큰 피해를 주는 주요 원인이다. 본 연구에서는 건강한 인삼에서 내생균을 분리하고, 인삼뿌리썩음병원균에 대해 길항작용을 갖는 내생균을 선발하였다. 분리된 17개 균주 중, 3개 균주가 인삼뿌리썩음병원균에 길항작용을 나타냈으며 인삼뿌리썩음병균이 생산하는 곰팡이독소인 라디시콜에 내성을 보였다. 우수한 길항효과와 라디시콜 저항성을 갖는 Bacillus velezensis CH-15를 선발하여 추가 실험을 수행하였다. 인삼 뿌리에 CH-15를 접종하면 병원균의 균사 생장을 억제할 뿐만 아니라 질병의 진행도 억제하였다. CH-15는 또한 바실로마이신 D, 이투린 A, 바실리신, 서팩틴 생합성 유전자를 갖고 있었다. 또한 CH-15 배양여액은 병원균 생장과 분생포자 발아를 억제하였다. 본 연구는 내생균 CH-15가 인삼뿌리썩음병 병원체에 대해 길항작용을 하고 인삼뿌리썩음병의 진행을 억제함을 보여주었으며 이 균주가 인삼뿌리썩음병을 억제하는 미생물 제제가 될 수 있을 것으로 기대한다.

Abstract ▼ AI-Helper

Ginseng is an important medicinal plant cultivated in East Asia for thousands of years. It is typically cultivated in the same field for 4 to 6 years and is exposed to a variety of pathogens. Among them, ginseng root rot is the main reason that leads to the most severe losses. In this study, endophytic bacteria were isolated from healthy ginseng, and endophytes with antagonistic effect against ginseng root rot pathogens were screened out. Among the 17 strains, three carried antagonistic effect, and were resistant to radicicol that is a mycotoxin produced by ginseng root rot pathogens. Finally, Bacillus velezensis CH-15 was selected due to excellent antagonistic effect and radicicol resistance. When CH-15 was inoculated on ginseng root, it not only inhibited the mycelial growth of the pathogen, but also inhibited the progression of disease. CH-15 also carried biosynthetic genes for bacillomycin D, iturin A, bacilysin, and surfactin. In addition, CH-15 culture filtrate significantly inhibited the growth and conidial germination of pathogens. This study shows that endophytic bacterium CH-15 had antagonistic effect on ginseng root rot pathogens and inhibited the progression of ginseng root rot. We expected that this strain can be a microbial agent to suppress ginseng root rot.

주제어

표/그림 (6)

그림 Fig. 1. Antifungal activity and radicicol resistance of bacteria isolated from healthy ginseng. (A) Antifungal activity of the bacterial strains, CH-9, CH-13, and CH-15, against ginseng root rot pathogens. CD, Cylindrocarpon destructans 14seo 01-01; FS, Fusarium solani. (B) Radicicol resistance of C. destructans KACC41077 and the three bacterial strains.
그림 Fig. 2. Effect of CH-15 culture filtrate on spore germination (A) and mycelial growth (B) of ginseng root rot pathogens. Fungal spore germination rates of CD-1 was determined in 4, 8, 12, and 24 h after incubation at 20°C, and FS was determined in 4, 8, and 12 after incubation at 25°C. Mock did not contain culture filtrate. CD, Cylindrocarpon destructans 14seo 01-01; FS, Fusarium solani. Asterisks indicate P-value (**P<0.01,***P<0.001) after comparison with t-test, and data are presented as mean±standard deviation.
표 Table 1. Primers used in this study
그림 Fig. 3. Maximum likelihood phylogenetic tree based on partial 16S rRNA (A) and gyrA (B) gene sequences from Bacillus strains. Bootstrap frequencies are shown above the branches.
그림 Fig. 4. Pathogenicity test of Panax ginseng and biosynthetic genes of antifungal metabolites. (A) Effect of CH-15 on ginseng root rot caused by Cylindrocarpon destructans 14seo 01-01. The ginseng 1-4 were inoculated with distilled water, CH-15, C. destructans, and both CH-15 and C. destructans 14seo 01-01, respectively. (B) The presence or absence of biosynthetic genes of antifungal metabolites in CH-15. L1, 100 bp DNA ladder; L2, zwiA (779 bp); L3, BmyA (875 bp); L4, ituA (647 bp); L5, bacD (749 bp); L6, srfA (441 bp); L7, fenD (964 bp); L8, 16S rRNA (1,511 bp); L9, λDNA/HindIII marker.
그림 Fig. 5. Antagonistic activity of CH-15 against ginseng pathogens. CD-1, Cylindrocarpon destructans 14seo 01-01; CD-2, C. destructans KACC 41077; SS, Sclerotinia sclerotiorum; FS, Fusarium solani; BC, Botrytis cinerea; AP, Alternaria panax.

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