[논문]인공접종을 통한 국내 야생 장미과 식물의 화상병 감수성 검정

박인웅; 송유림; 오엄지; 김요엘; 황인선; 전미진; 안초롱; 김진숙; 김순옥; 오창식

doi:10.5423/rpd.2023.29.1.23

인공접종을 통한 국내 야생 장미과 식물의 화상병 감수성 검정
Determination of Fire Blight Susceptibility on Wild Rosaceae Plants in Korea by Artificial Inoculation 원문보기

Research in plant disease = 식물병연구, v.29 no.1, 2023년, pp.23 - 38

박인웅 (서울대학교 식물면역연구센터) , 송유림 (경희대학교 원예생명공학과) , 오엄지 (경희대학교 그린바이오과학원) , 김요엘 (경희대학교 그린바이오과학원) , 황인선 (경희대학교 원예생명공학과) , 전미진 (국립생물자원관 미생물자원과) , 안초롱 (국립생물자원관 미생물자원과) , 김진숙 (한반도식물다양성연구소) , 김순옥 (국립생물자원관 미생물자원과) , 오창식 (서울대학교 식물면역연구센터)

초록
AI-Helper

Erwinia amylovora에 의해 발생하는 화상병은 2015년 국내에서 처음 보고된 이후 전국적으로 확산되고 있다. 화상병은 벌과 같은 화분매개충, 강한 바람을 동반한 강수와 이병주 전정 작업에 의한 전파 등 많은 경로를 통해 병원균 감염이 진행되어 발생한다. 이 외에도 자생하는 개야광나무속, 산사나무속, 피라칸다속, 벚나무속, 마가목속 등의 장미과 식물이 E. amylovora의 기주식물과 전파원이 될 수 있음이 보고되어 있다. 그러나, 국내에서는 아직까지 자생하고 있는 장미과 식물의 화상병 감수성에 관한 연구가 많이 다뤄지지 않은 상황이다. 본 연구에서는 2020년부터 2022년까지 3년간 국내에서 채집한 야생 장미과 식물 10속 25종을 대상으로 화상병 감수성 검정을 진행하였다. 10⁸ cfu/ml 농도의 E. amylovora 균액을 이용하여 채집한 식물의 꽃, 잎과 가지, 과실에 접종하고, 화상병 유사 병징 발현을 관찰하고 PCR을 통해 접종 조직 내의 E. amylovora 검출 여부를 분석하였다. 그 결과, 병징 발현과 병원균 검출 결과가 동일한 14종의 야생 장미과 식물을 확인하였다. 이러한 결과는 향후 적절한 화상병 방제 정책 수립에 기여할 수 있을 것으로 생각된다.

Abstract ▼ AI-Helper

The fire blight caused by Erwinia amylovora (Ea) is a devastating disease of Rosaceae plants, including commercially important apple and pear trees. Since the first report in Korea in May 2015, it has been spreading to neighboring regions gradually. Host plants can be infected by pollinators like bees, rainfall accompanied by wind, and cultural practices such as pruning. Many studies have revealed that wild Rosaceae plants such as Cotoneaster spp., Crataegus spp., Pyracantha spp., Prunus spp., and Sorbus spp. can be reservoirs of Ea in nature. However, wild Rosaceae plants in Korea have not been examined yet whether they are susceptible to fire blight. Therefore, the susceptibility to fire blight was examined with 25 species in 10 genera of wild Rosaceae plants, which were collected during 2020-2022, by artificial inoculation. Bacterial suspension (108 cfu/ml) of Ea type strain TS3128 was inoculated artificially in flowers, leaves, stems, and fruits of each plant species, and development of disease symptoms were monitored. Moreover, the presence of Ea bacteria from inoculated samples were checked by conventional polymerase chain reaction. Total 14 species of wild Rosaceae plants showed disease symptoms of fire blight, and Ea bacteria were detected inside of inoculated plant parts. These results suggest that wild Rosaceae plants growing nearby commercial apple and pear orchards in Korea can be Ea reservoirs, and thus they should be monitored regularly to minimize the damage by Ea infection and spreading.

주제어

표/그림 (12)

그림 Fig. 1 . Geographical location of wild Rosaceae plants collected in Korea from 2020 to 2022. The blue, red, and green stars indicate the locations of collected plant samples in 2020, 2021, and 2022, respectively. The map was obtained from the National Geographic Information Institute (NGII) in Korea.
표 Table 1. List of wild Rosaceae plants in Korea used in this study during 2020–2022
그림 Fig. 2. The scheme of the inoculation methods to confirm a fire blight susceptibility in wild Rosaceae plants. (A) The inoculation on the flower. The flowers were cut from the pedicel and placed into 2 ml of 10% sucrose solution, and then 20 μl of bacterial suspension (10⁸ cfu/ml of Erwinia amylovora TS3128) was dropped on the stigma. In 2022, the method was modified, so that the flower clusters were detached from the twig and placed into 4 ml of sterilized tap water. (B) The inoculation on the leaf and twig. Young twigs with leaves were soaked in 4 ml of sterilized tap water. Leaves were cut in a diagonal direction with a scissor, which was sufficiently soaked in the bacterial suspension. (C) The inoculation on the fruit. Fruits were punched to make a hole by pipette tip or needle of a syringe. The 20 μl of bacterial suspension was dropped into the hole. All inoculated samples were kept in insulated moist plastic boxes at 26°C.
표 Table 2. Summary of disease symptom development in wild Rosaceae plants in Korea after inoculation of Erwinia amylovora
그림 Fig. 3. Screening of the fire blight susceptibility in the genera Chaenomeles, Cotoneaster, and Crataegus. All plants samples were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. The white and blue arrows indicate the bacterial ooze on the Chaenomeles speciosa surface and discoloration on the pedicel of Crataegus pinnatifida and Crataegus scabrida by Ea infection, respectively. The samples showing suspicious disease symptoms were remarked by the red boxes. The photos were taken 8 to 11 days after inoculation. ND, not determined.
표 Table 3. Summary of the PCR results to detect Erwinia amylovora in the inoculated parts of wild Rosaceae plants in Korea
그림 Fig. 4. Detection of Erwinia amylovora in both inoculated and non-inoculated parts of the wild Rosaceae plants by conventional polymerase chain reaction (PCR). E. amylovora TS3128 was detected from inside of several inoculated samples. The bands between 900 bp to 1.1 kb corresponded the PCR product of E. amylovora pEA29 by a specific primer set. M, the mock with 10 mM MgCl2; L, leaf samples; S, stem samples; P, E. amylovora TS3128 as a positive control; N, sterilized distilled water as a negative control.
그림 Fig. 5. Screening of the fire blight susceptibility in the genus Malus. All plant samples were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. Leaves and twigs displayed clear disease symptoms than other plant parts. The samples showing suspicious disease symptoms were remarked by the red boxes. The samples were photographed 10 to 11 days after inoculation.
그림 Fig. 6. Screening of the fire blight susceptibility in the genus Pyrus. All plant samples were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. Flowers of Pyrus ussuriensis and fruits of Pyrus ussuriensis var. hondoensis were not tested. The samples showing suspicious disease symptoms were remarked by the red boxes. The photos were taken 10 to 11 days after inoculation. ND, not determined.
그림 Fig. 7. Screening of the fire blight susceptibility in the genus Prunus. All twigs and fruits were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. The samples, showing suspicious disease symptoms, were remarked by the red boxes. The photos were taken 10 to 11 days after inoculation.
그림 Fig. 8. Screening of the fire blight susceptibility in the genera Pyracantha, Rosa, and Rubus. All plant samples were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. Flowers of the genera Rosa and Rubus were not examined in this experiment. The samples showing suspicious disease symptoms were remarked by the red boxes. The samples were photographed 10 to 11 days after inoculation. ND, not determined.
그림 Fig. 9. Screening of the fire blight susceptibility in the genera Sorbus. All plant samples were inoculated with 10⁸ cfu/ml of Erwinia amylovora (Ea) TS3128. Flowers of the genera S. commixta and S. ulleungensis were not examined in this experiment. The samples showing suspicious disease symptoms were remarked by the red boxes. The samples were photographed 7 to 11 days after inoculation (dai). ND, not determined.

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