Division of Applied Bioscience and Biotechnology, Institute of Environmentally-Friendly Agriculture (IEFA), College of Agricultural and Life Science, Chonnam National University, Gwangju 500-757, Republic of Korea.
Microb Pathog. 2013 Jun-Jul;59-60:52-9. doi: 10.1016/j.micpath.2013.04.005. Epub 2013 Apr 18.
In this study, the 3,4-dihydroxybenzoic acid (3,4-DHBA) from Terminalia nigrovenulosa bark (TNB) was purified and its in vitro nematicidal activity was investigated against Meloidogyne incognita. The purification of 3,4-DHBA used a silica gel column and Sephadex LH-20 chromatography combined with thin-layer chromatography and high performance liquid chromatography. Structural identification of the 3,4-DHBA was conducted using (1)H nuclear magnetic resonance (NMR), (13)C NMR, and liquid chromatography time-of-flight mass spectrometry. Nematicidal activity bioassays revealed that 3,4-DHBA treatment resulted in 33.3, 47.5, 72.5 and 94.2% J2 mortality at 0.125, 0.25, 0.5 and 1.0 mg/ml, respectively after 12 h incubation. J2 mortality was increased significantly (P < 0.0001) with increasing incubation time in the range of 54.2-94.2% from 3 to 9 h after incubation with 3,4-DHBA (1.0 mg/ml), but with no significant difference observed where the incubation time was increased from 9 to 12 h. The 3,4-DHBA treatment resulted in 33.3, 65.0, 76.7 and 85.0% hatch inhibition at 0.125, 0.25, 0.5 and 1.0 mg/ml, respectively, 3 days after incubation. Changes in the shape of the eggs were determined after incubation for 1 day with a 3,4-DHBA concentration of 1.0 mg/ml.
在这项研究中，从诃子（Terminalia nigrovenulosa bark，TNB）树皮中纯化出 3,4-二羟基苯甲酸（3,4-DHBA），并研究了其对南方根结线虫（Meloidogyne incognita）的体外杀线虫活性。3,4-DHBA 的纯化采用硅胶柱和 Sephadex LH-20 层析结合薄层层析和高效液相色谱法。通过（1）H 核磁共振（NMR）、（13）C NMR 和液质联用飞行时间质谱对 3,4-DHBA 的结构进行了鉴定。杀线虫活性生物测定表明，在 12 h 孵育后，0.125、0.25、0.5 和 1.0 mg/ml 浓度的 3,4-DHBA 处理分别导致 J2 死亡率为 33.3%、47.5%、72.5%和 94.2%。在 3-9 h 孵育期间，随着 3,4-DHBA（1.0 mg/ml）浓度的增加，J2 死亡率显著增加（P<0.0001），从 54.2%增加到 94.2%，但在 9-12 h 孵育时间增加时，没有观察到显著差异。在 3 天后，3,4-DHBA 处理在 0.125、0.25、0.5 和 1.0 mg/ml 浓度下分别导致孵化抑制率为 33.3%、65.0%、76.7%和 85.0%。在 1.0 mg/ml 3,4-DHBA 浓度下孵育 1 天后，观察到卵的形状发生变化。
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