| 林可竟 1#, 周 挺 2#, 朱永平 2, 顾 钢 2*, 唐雨晴 1, 谢雪金 1, 蔡学清 1*.红色荧光蛋白(RFP)标记法揭示烟草野火病菌侵染根系的新途径[J].植物保护,2026,52(4):88-99. |
| 红色荧光蛋白(RFP)标记法揭示烟草野火病菌侵染根系的新途径 |
| Red fluorescent protein (RFP) labeling reveals a novel root invasion pathway of Pseudomonas amygdali pv. tabaci in tobacco seedlings |
| 投稿时间：2025-08-22 修订日期：2026-01-07 |
| DOI：10.16688/j.zwbh.2025438 |
| 中文关键词: 烟草野火病菌 云烟87 根部侵入途径 激光共聚焦显微镜 红色荧光蛋白基因 |
| 英文关键词:Pseudomonas amygdali pv. tabaci Yunyan 87 root invasion pathway confocal laser scanning microscope red fluorescent protein gene (rfp) |
| 基金项目:中国烟草总公司福建省公司科技计划项目(2022350000240071);中国烟草总公司科技计划项目(110202201028［LS-12］) |
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| 中文摘要: |
| 为明确烟草野火病病原菌扁桃假单胞菌烟草致病变种Pseudomonas amygdali pv. tabaci的根侵入途径, 本研究采用浸根法接种2片真叶和5片真叶的烟苗, 通过激光共聚焦显微镜观察烟草野火病菌的侵染定殖情况;并采用灌根法接种烟苗, 通过稀释分离法分离并统计烟株根、茎、叶内的病原菌数量。结果表明, 浸根接种后24 h, 在烟苗侧根与主根的根尖伸长区、根毛区以及侧根与主根连接处均聚集烟草野火病菌, 同时在根和茎的维管束细胞内也可观察到病原菌;接种后1~10 d, 根表菌量逐渐下降, 而根组织内的菌量则先上升后下降。根表菌量在接种后1 d达到峰值, 为5.00×10^6 cfu/g;根组织内的菌量在接种后5 d达到峰值, 为3.91×10^6 cfu/g。灌根接种后可在烟株的根、茎、叶内分离回收到接种的病原菌。另外, 病田的无症状及发病烟草植株的根、茎、叶均可分离到烟草野火病菌, 菌量为7.25×10^2~5.41×10^5 cfu/g。研究结果揭示了烟草野火病菌新的侵入途径, 即从根部侵入, 并通过维管束传导从而引发叶片坏死, 该侵入途径的证实为野火病的防治提供了新思路。 |
| 英文摘要: |
| To clarify the root invasion pathway of the tobacco wildfire pathogen Pseudomonas amygdali pv. tabaci, tobacco seedlings at the two-true-leaf and five-true-leaf stages were inoculated by the root-dip method, and the infection and colonization of the pathogen were observed using confocal laser scanning microscopy. In addition, tobacco seedlings were inoculated by the root-irrigation method, and the number of pathogens in roots, stems, and leaves was isolated and quantified using the dilution plating method. The results showed that 24 h after root-dip inoculation, P.amygdali pv. tabaci accumulated in the root tip elongation zone, root hair zone, and the junction between lateral roots and taproot. Meanwhile, the pathogen could also be observed inside the vascular bundle cells of roots and stems. From one to ten days after inoculation, the bacterial population on the root surface gradually declined, whereas the bacterial population inside root tissues initially increased and then decreased. The bacterial population on the root surface reached a peak of 5.00×10 6 cfu/g on day one after inoculation, while the bacterial population inside root tissues reached a peak of 3.91×10 6 cfu/g on day five after inoculation. The inoculated pathogen could also be re-isolated from the roots, stems, and leaves of tobacco plants after root-irrigation inoculation. In addition, P.amygdali pv. tabaci was isolated from the roots, stems, and leaves of both asymptomatic and diseased tobacco plants in diseased fields, with bacterial populations ranging from 7.25×10 2 to 5.41×10 5 cfu/g. These findings revealed a novel invasion pathway of the tobacco wildfire pathogen, namely root invasion followed by vascular bundle transmission, which eventually leads to leaf necrosis. Confirmation of this invasion pathway provides new insights for the prevention and control of tobacco wildfire disease. |
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