信息摘要：
2026年，赣南师范大学江西省特色园艺植物病虫害防控重点实验室团队在《Microorganisms(ISSN 2076-2607)》(SCI期刊，MDPI出版社)发表文献《Bacillus velezensis RF2 Rescued from Citrus Phyllosphere: Dual Mechanisms and Broad-Spectrum Activity for Controlling Citrus Bacterial Canker》，文献中使用LUYOR-3415RG双波长荧光蛋白激发光源对绿色荧光蛋白标记的柑橘溃疡病菌(Xcc-GFP)进行荧光激发与成像观察，用于生防菌高通量筛选、抑菌活性评估及叶片活体防效评价，为柑橘溃疡病(CBC)绿色防控菌株的筛选及其作用机制研究提供关键实验证据。
赣南师范大学将LUYOR-3415RG用于Xcc-GFP荧光筛选与柑橘溃疡病防控研究
2026年，赣南师范大学江西省特色园艺植物病虫害防控重点实验室团队在《Microorganisms(ISSN 2076-2607)》(SCI期刊，MDPI出版社)发表文献《Bacillus velezensis RF2 Rescued from Citrus Phyllosphere: Dual Mechanisms and Broad-Spectrum Activity for Controlling Citrus Bacterial Canker》，文献中使用LUYOR-3415RG双波长荧光蛋白激发光源用于Xcc-GFP荧光成像实验，对绿色荧光蛋白标记的柑橘溃疡病菌进行荧光激发与观察，用于拮抗菌株高通量筛选、抑菌活性评估以及叶片活体防效评价。LUYOR-3415RG双波长荧光蛋白激发光源在实验流程中承担关键的荧光激发与成像环节，通过特定波长激发光使Xcc-GFP发出明亮绿色荧光，实现抑菌圈、接种叶片等样品的直观成像，为研究结果的可重复性与可靠性提供保障。
赣南师范大学位于江西赣州，是集师范教育、科学研究与社会服务于一体的省属重点大学，也是江西省特色园艺植物病虫害防控重点实验室的依托单位，长期面向赣南脐橙等特色园艺作物开展病虫害绿色防控研究。柑橘溃疡病(CBC)由柑橘黄单胞杆菌柑橘亚种(Xanthomonas citri subsp. citri，Xcc)引起，是威胁柑橘产业可持续发展的重大病害之一。该研究团队以“植物求救(Cry for Help)"假说为指导，从自然感病柑橘叶际分离获得72株细菌，利用Xcc-GFP荧光筛选方法系统筛选出高效拮抗菌株贝莱斯芽孢杆菌RF2(Bacillus velezensis RF2，Bv-RF2)，并通过全基因组测序、antiSMASH基因簇预测、代谢组学分析及寄主防御基因表达分析，揭示了Bv-RF2通过分泌化学性质稳定的活性成分并诱导寄主ISR样防御“预激(priming)"双重机制抑制Xcc的分子基础。LUYOR-3415RG用于的Xcc-GFP荧光成像作为筛选拮抗菌株、评估抑菌活性与活体防效的关键手段，为上述研究提供了重要实验证据。
本实验采用LUYOR-3415RG双波长荧光蛋白激发光源作为Xcc-GFP荧光成像的核心激发光源，用于对绿色荧光蛋白标记的柑橘溃疡病菌进行荧光激发与成像观察。生防菌筛选阶段，将叶际分离菌株与Xcc-GFP共培养，经LUYOR-3415RG激发光源辐照后观察荧光信号强弱，快速判定菌株的拮抗活性，最终获得抑菌率高达78.89%的高效菌株Bv-RF2；抑菌活性评估阶段，将Bv-RF2发酵产物甲醇粗提物(20、10、5、2.5、1 mg/mL)点样于接种Xcc-GFP的NA平皿，28 ℃培养36-48 h后经LUYOR-3415RG照明成像，清晰显示不同浓度处理产生的抑菌圈(Fig. 2A)，其中1 mg/mL粗提物抑菌圈直径为11.87±0.55 mm，大于0.1 mg/mL硫酸链霉素阳性对照的9.76±0.09 mm；叶片活体防效评价阶段，将Xcc-GFP(OD600=0.6)与不同浓度粗提物按1:1混合接种“哈姆林"甜橙叶片，7 dpi时在LUYOR-3415RG照明下采集GFP荧光图像(Fig. 5A)，直观显示10、5、1 mg/mL粗提物处理分别使病斑面积减少75.93%、43.32%和27.98%。LUYOR-3415RG双波长荧光蛋白激发光源用于Xcc-GFP荧光成像结果表明，该激发光源可稳定激发绿色荧光蛋白发光，荧光信号清晰、成像对比度高，为生防菌高通量筛选、抑菌活性定量评估及活体防效直观评价提供了可靠的技术手段，为揭示Bv-RF2防控柑橘溃疡病的双重机制提供了直接实验证据。
图1 LUYOR-3415RG照明下Xcc-GFP抑菌圈成像（论文Fig. 2A）
该研究证实，LUYOR-3415RG双波长荧光蛋白激发光源可用于Xcc-GFP荧光成像实验，适用于荧光标记病原菌检测、拮抗菌高通量筛选、抑菌活性评估与活体防效荧光成像等微生物学与植物病理学应用场景。LUYOR-3415RG双波长荧光蛋白激发光源在国内外高校和科研院所得到广泛好评，欢迎私信或留言联系上海路阳了解详情！
原文段落
Xcc-GFP荧光成像方法原文
Inhibition zones formed by methanol extracts at different concentrations against Xcc-GFP on NA medium were imaged under LUYOR-3145RG illumination. 1 to 6: the concentrations of the methanol extract: 20 mg/mL, 10 mg/mL, 5 mg/mL, 2.5 mg/mL, 1 mg/mL and streptomycin (Strep, as a positive control, with a working concentration of 0.1 mg/mL), 7: ddH2O. Log-phase Xcc-GFP cells were scraped from NA plates, washed three times with PBS, and resuspended to OD600 = 1.0. Aliquots (0.1 mL) were mixed with 10 mL of molten 1.2% NA, poured into Petri dishes, and allowed to solidify. Then, 5 µL of each test solution was spotted onto the surface: Bv-RF2 crude methanol extract (10 mg/mL), surfactin (10 µM, C53H93N7O13) (TargetMol, Boston, MA, USA), streptomycin sulfate (0.1 mg/mL), and PBS (control). Plates were incubated at 28 °C for 36–48 h, and inhibition zones were measured using ImageJ (version 1.54f, Wayne Rasband, NIH, Bethesda, MD, USA).
Xcc-GFP筛选与活体防效结果原文
Using an Xcc-GFP-based screening method, strain Bv-RF2 exhibited strong antagonism toward Xcc. Under the screening conditions, the fermentation broth of Bv-RF2 achieved an inhibition rate of 78.89%. Co-inoculation of Xcc with increasing concentrations of the extract resulted in a clear, dose-dependent reduction in lesion area (Figure 5A,B). Quantitatively, extracts at 10, 5, and 1 mg/mL reduced disease severity by 75.93%, 43.32%, and 27.98% (Figure 5C).
研究结论原文
In conclusion, we identified Bacillus velezensis RF2 (Bv-RF2) from the phyllosphere of Xcc-infected citrus and demonstrated its inhibitory activity against Xcc as well as other citrus-associated pathogens under our experimental conditions. Crude methanolic extracts exhibited strong antibacterial activity in vitro and reduced canker development in a leaf assay, indicating that Bv-RF2-derived soluble components can contribute to disease suppression. Genome mining revealed a repertoire of predicted NRPS/PKS-type BGCs that is consistent with the capacity to produce diverse secondary metabolites; however, causal links between specific clusters/metabolites and antimicrobial activity require further targeted validation. Bv-RF2 infiltration was associated with an ISR-like, priming-related host response characterized by transient induction of defense-related genes. Future work should evaluate live-bacterium foliar biocontrol performance, including colonization and persistence, and apply targeted chemical and genetic approaches to strengthen genotype–phenotype relationships.
图2 LUYOR-3415RG照明下甜橙叶片活体防效GFP荧光成像（论文Fig. 5A）
图3 病斑面积与抑制率定量分析（论文Fig. 5B、5C）
DOI:10.3390/microorganisms14010121
