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森林假单胞菌抑菌物稳定性及其成分分析
梁文洪,赵倩,张良军,赖国栋,凌斯全,何静,徐金柱,田龙艳
1.德庆县森林病虫害防治检疫站;2.甘肃农业大学;3.广东省林业科学研究院;4.广东省森林培育与保护利用重点实验室/ 广东省林业科学研究院
摘要:
森林假单胞菌 Pseudomonas silvicola 菌株 T1-3-2 对杉木炭疽病具有较强的防治效果, 为将其研 发为林木炭疽病的生防制剂, 需进一步探明该菌抑菌谱、 抑菌物质稳定性以及主要成分等, 以补充完善该 菌的生防理论。 文章通过平板对峙法、 菌丝生长速率法、 液相色谱/ 线性离子阱-静电场轨道阱高分辨质谱 法 (UPLC/ LTQ-Orbitrapa MS) 和气相色谱-质谱法 (GC-MS) 分析了该菌的抑菌谱、 抑菌物质的稳定性 以及主要成分。 结果表明, 菌株 T1-3-2 对 9 株供试病原真菌均具有较好的抑菌活性, 尤其对包括杉木炭 疽病菌 Colletotrichum gloeosporioides 在内的多种林木炭疽病菌表现出较强的抑菌作用, 抑制率均达到 65% 以上; 其代谢产物粗提物对杉木炭疽病菌的孢子萌发和菌丝生长具有明显抑制作用; 温度≤70 ℃、 酸碱度以及紫外辐射对粗提物抑菌效果均无显著影响, 但高温处理 ( T > 70 ℃) 对抑菌效果影响显著; 粗提物主要含有 14 种化合物, 其中包含假单胞菌常见的 4 种吩嗪类抑菌物质, 即吩嗪、 1, 6-二羟基吩 嗪、 10-乙酰基-3, 7-二羟基吩嗪和绿脓菌素; 挥发性气体 (VOCs) 中包含: 苯类、 烯烃类、 酮类等 13 种化合物。 菌株 T1-3-2 具广谱抑菌特性, 其粗提物稳定性较好且具较强抑菌活性, 该菌株及其代谢物在 林木炭疽病的防治中有很好的应用前景。
关键词:  森林假单胞菌  抑菌物质  稳定性
DOI:
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基金项目:广东省林业科技创新项目 (2023KJCX020), 国家自然科学基金 (32060341)。
Stability and Composition Analysis of Antifungal Substances of Pseudomonas silvicola
liangwenhong1, zhaoqian2, zhangliangjun3, LAI GUO DONG4, LING SI QUAN4, HE JING5, XU Jinzhu6, tian long yan7
1.Forest Pest Control and Quarantine Station of Deqing County;2.Gansu Agricultural University;3.GUANGDONG ACADEMY OF FORESTRY;4.Guangdong Academy of Forestry;5.GanSu angricutrual university;6.Guangdong Provincial Key Laboratory of Silviculture, Protection and Utilization / Guangdong Academy of Forestry;7.Guang Dong Academy of Forestry
Abstract:
Pseudomonas silvicola T1-3-2 has stronger antagonistic effects on Chinese fir anthracnose. It is necessary to explore its inhibition spectrum, antifungal activity, and stability to implement the utilization and improve its biocontrol theory. In this study, its inhibition spectrum, the stability, and the main components of the antifungal substance were analyzed by the plate confrontation method, mycelium growth rate method, and UPLC/ LTQ-Orbitrapa MS and GC-MS. The results showed that T1-3-2 had stronger antifungal activity against 9 pathogenic fungi, especially with an inhibition rate of more than 65% against anthracnose of Cunninghamia lanceolata and other trees. The crude extracts had a significant inhibitory on the spore germination and mycelial growth of C. gloeosporioides. They were substantially stable after the different treatments with pH, ultraviolet light, and temperature below 70 ℃ in the antifungal tests. The crude extract of strain T1-3-2 contains 14 compounds, including 4 kinds of phenazine and its derivatives ( phenoxazine, 1, 6-dihydroxyphenazine, 10-acetyl-3, 7-dihydroxyphenazine, pyocyanin) of main antifungal substances reported in Pseudomonas. Volatile gases (VOCs) contain 13 compounds such as benzene, olefins, ketones, and so on. The strain T1-3-2 has broad-spectrum antifungal properties, and its crude extract had substantial stability and stronger antifungal activity, it shows that the strain and its metabolites have good application prospects in the prevention and control of forest anthracnose.
Key words:  Pseudomonas silvicola  Antifungal substance  Stability