Effects of Penicillium oxalicum C11 on Rehmannia glutinosa growth and its metabolites analysis

doi:10.17521/cjpe.2022.0439

Abstract

Abstract:

Aims The aim of this study was to isolate the harmful strain and analyze the effects of its metabolites on Rehmannia glutinosa, which provided theoretical support for solving R. glutinosa continuous cropping obstacles.

Methods The fungi were isolated from continuous cropping soil. The fungal fermentation liquid metabolites were separated using different polarity gradient extraction and silica gel column chromatography. Metabolites were determined by high performance liquid chromatography and liquid chromatography-mass spectrometry.

Important findings A strain of Penicillium oxalicum C11 was isolated from continuous cropping soil. The toxic metabolite of P. oxalicum C11 was identified as cyclo (L-Arg-L-Leu) dipeptide (molecular weight: 269.347, molecular formula: C₁₂H₂₃N₅O₂). Results indicated P. oxalicum C11 inhibited R. glutinosa yield by 41.5%, suggesting that the toxic metabolite L-Arg-L-Leu could be vital for R. glutinosacontinuous cropping obstacles.

Key words: Rehmannia glutinosa, continuous cropping obstacles, mold, Penicillium oxalicum

HAO Shang-Hua, LUO Meng-Xiang, CAO Hong-Li, ZHANG Sen, WANG Ming-Dao. Effects of Penicillium oxalicum C11 on Rehmannia glutinosa growth and its metabolites analysis[J]. Chin J Plant Ecol, 2024, 48(6): 809-816.

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URL: https://www.plant-ecology.com/EN/10.17521/cjpe.2022.0439

https://www.plant-ecology.com/EN/Y2024/V48/I6/809

Figures/Tables 4

Table 1 Parameters and settings of high-performance liquid chromatography (HPLC) and electrospray liquid chromatography-mass spectrometry (LC-MS)

HPLC	LC-MS
色谱柱 Chromatographic column: ACQUITY UPLC HSS T3	色谱柱 Chromatographic column: ACQUITY UPLC HSS T3
孔径 Aperture: 5 μm	进样量 Sample quantity: 2 μL
进样量 Sample quantity: 15 μL	流速 Velocity: 0.5 mL·min^-1
流动相: A, 0.1%甲酸乙腈; B, 纯水	柱温 Column temperature: 30 ℃
Mobile phase: A, 0.1% formic acid acetonitrile; B, pure water	流通池路径长度 Flow pool path length: 10.000 mm
流速 Velocity: 0.5 mL·min^-1	流通池体积 Flow cell volume: 0.500 μL
柱温 Column temperature: 40 ℃	扫描范围 Scan range: 50-1500 m/z
检测波长 Detection wavelength: 500 nm	低压极限 Low pressure limit: 0 pounds per square inch
进样时间 Sample time: 16.5 min	高压极限 High pressure limit: 15 000 pounds per square inch

Fig. 1 Screening of harmful strains and effects of Penicillium oxalate C11 strain on the growth of Rehmannia glutinosa (mean ± SD). A, Seedling morphology of R. glutinosa. B, Mass of R. glutinosa root. C, Length of R. glutinosa root. D, Effect of fermentation broth of C11 strain on callus of R. glutinosa. E, Quality of potted R. glutinosa. F, R. glutinosa morphology of different treatments. They are first crop R. glutinosa, C11 strain treated R. glutinosa, and continuous crop R. glutinosa, respectively, from left to right. G, Mass of field R. glutinosa. CK, control; C7, C11, F2, F9, F11, strains. Different lowercase letters indicate significant differences (p < 0.05). ** p < 0.01; ***, p < 0.001.

Fig. 2 Penicillium oxalicum C11 strain morphology and phylogenetic tree. A, Positive side of C11 colony. B, Reverse side of C11 colony. C, C11 mycelium and spore morphology. D, Phylogenetic tree.

Fig. 3 Biomeasurement results of different polar extraction phases and silica gel column chromatography of Penicillium oxalicum fermentation broth. A, Bioassay results of different phases (mean ± SE). B, Bioassay results of collected pipes with water phases 51-59 (mean ± SE). C, Statistics of biological test results of R. glutinosa seedlings (mean ± SE). D, First-order mass spectrogram of the material peak. E, Secondary mass spectrogram of the material peak. F, Liquid phase diagram of pure substance and 54 sample tube. CK, control. Different lowercase letters indicate significant differences (p < 0.05).

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