朱砂莲果实化学成分杀南方根结线虫活性研究

为了解朱砂莲(Aristolochia tuberosa O.F.Liang et S.M.Huang)的化学成分,从其果实的甲醇提取物中分离得到4个化合物。通过波普数据分析,分别鉴定为马兜铃内酰胺W(1)、8-epidiosbulbin e acetate(2)、diosbulbin B(3)和β-sitosterol(4)。化合物1~3对南方根结线虫2龄幼虫具有不同程度的毒杀作用,尤其马兜铃内酰胺W(1)活性最好,其96 h后的LC50为119.94μg m L–1。马兜铃属植物具有开发为新型植物源杀根结线虫剂的潜在价值。     

Nematicidal activity against Aphelenchoides besseyi and Ditylenchus destructor of three biflavonoids,isolated from roots of Stellera chamaejasme

Aphelenchoides besseyi and Ditylenchus destructor can cause serious problems for a number of important agricultural crops and vegetables. In this study, the ethanol extract of Stellera chamaejasme L. roots showed strong nematicidal activity against Aphelenchoides besseyi and Ditylenchus destructor. By using a bioactivity-driven fractionation, three biflavonoids were isolated from the extract and their structures were identified by mass and nuclear magnetic resonance spectral data. Nematicidal activity bioassays revealed that isoneochamaejasmin A had the strongest nematicidal activity against A. besseyi and D. destructor with LC₅₀ values of 2.32 and 0.18?mM at 72?h, respectively. Chamaejasmenin B displayed weaker nematicidal activity against A. besseyi with an LC₅₀ value of 3.94?mM at 72?h. Neochamaejasmin B induced the lowest mortality against D. destructor with an LC₅₀ values of 15.6?mM at 72?h. These results suggested that the kind and position of substitutions and the relative configuration of 2-H/3-H and 2”-H/3”-H could be considered as important factors responsible for the nematicidal activity of these purified C-3/C-3″ biflavonoids.     

Stereumin A-E, sesquiterpenoids from the fungus Stereum sp. CCTCC AF 207024

Five cadinane sesquiterpenoids, named stereumin A (1), B (2), C (3), D (4) and E (5) were isolated from the CHCl(3) extract of the culture broth of the fungal strain CCTCC AF 207024. Based on the sequences at the internal transcribed spacer (ITS) region and partial 28S rDNA, this fungus was identified as a Stereum sp. The structures of the five compounds were elucidated using spectroscopic data from 1D, 2D NMR and HRESIMS experiments, and the structures of 1 and 2 were further confirmed by single-crystal X-ray diffraction analysis. Compounds 1-5 showed nematicidal activities against the nematode Panagrellus redivivus at 400 mg l(-1). Among these five compounds, compounds 3 and 4 killed 84.4% and 94.9% of P. redivivus, respectively in 48 h.     

Purification and cloning of a novel serine protease from the nematode-trapping fungus <Emphasis Type="Italic">Dactylellina varietas</Emphasis> and its potential roles in infection against nematodes

From the culture filtrate of the fungus Dactylellina varietas (syn. Dactylella varietas), an extracellular protease (designed Dv1) was purified by cation exchange and hydrophobic interaction chromatography. The purified protease showed a molecular mass of approximately 30 kDa and displayed an optimal activity at pH 8 and 60.5°C (more than 20 min). This protease could degrade a broad range of substrates including casein, gelatin, BSA (bovine serum albumin), and nematode cuticle. However, its proteolytic activity was highly sensitive to the serine protease inhibitor Phenylmethylphonylfuoride (1 mM), indicating that it belongs to the serine-type peptidase group. This protease could immobilize the free-living nematodes Panagrellus redivivus and Caenorhabditis elegans and hydrolyze the purified cuticle of P. redivivus, suggesting it may play a role in infection against nematodes. The encoding gene of Dv1 and its promoter sequence were cloned using degenerate primers and the DNA walking technology. Its open-reading frame contains 1,224 base pairs and without any intron. The deduced amino-acid sequence shared low identity to serine proteases from other nematode-trapping fungi. Our report identified a novel pathogenic protease from the nematode-trapping fungus D. varietas, and the three-dimensional structure of this protease was predicted using the Swiss-Prot method. Jinkui Yang and Lianming Liang contributed equally to this work.     

Synthesis and nematicidal activities of 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea against Meloidogyne incognita

Two series of novel 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea were designed and synthesized. The bioassay results showed that most of the test compounds showed good nematicidal activity against M. incognita at the concentration of 10.0 mg L^?1in vivo. The compounds A13, A17 and B3 showed excellent nematicidal activity on the second stage juveniles of the root-knot nematode with the inhibition rate of 51.3%, 58.3% and 51.3% at the concentration of 1.0 mg L^?1 respectively. It suggested that the structure of 1,2,3-benzotriazin-4-one derivatives containing thiourea and acylthiourea could be optimized further.     

Biopesticidal value of selected essential oils against pathogenic fungus, termites, and nematodes

The biopesticidal potential of six plant-derived essential oils (mint [Mentha arvensis], ajwain [Carum capticum], lemongrass [Cymbopogon citrates], clove [Eugenia caryophyllata], cedarwood [Cedrus deodara], and eucalyptus [Eucalyptus globulas]) was evaluated against Odontotermes obesus (termites), Fusarium oxysporum (plant pathogenic fungi), and Meloidogyne incognita (nematodes). In the case of termites, a “no-choice” bioassay revealed that the mint oil gave the best results (100% mortality in 30 min with 10% oil and in 10 h with 0.12% oil) followed by the lemongrass and ajwain oils. The disc diffusion method was adopted to test the anti-fungal activity of the essential oils and it was found that the clove oil gave the maximum inhibition measured in terms of the average inhibition zone diameter (5.3 ± 0.2 cm with 10% oil and 6.6 ± 0.9 cm with 20% oil), followed by the ajwain oil. To check the anti-nematicidal activity of the essential oil, in-vitro growth chamber experiments revealed that eucalyptus oil was the most efficient (100% mortality in 6 h with 1000 ??l l⁻¹ oil and in 30 h with 125 ??l l⁻¹ oil), followed by the ajwain oil. The use of the crude oils at low concentrations provided satisfactory results at the laboratory level against these pathogens, and needs further evaluation in field trials.     

核苷N9705的抗病毒、抗菌杀虫活性研究

由一株链霉菌产生的核苷N970 5是肽酰胞嘧啶核苷衍生物 ,具有广谱抗生活性。N970 5能够抑制疱疹病毒I型 (HSV I)对Vero细胞、烟草花叶病毒 (TMV)对烟叶细胞的病理损伤作用 ;对四种水稻病原真菌和一种烟草病原真菌有较强的抑制生长作用 ;还对腐生线虫 (P .redivivus)及松材线虫 (B .xylophilus)有杀伤作用。     

Bioactive saponins from Microsechium helleri and Sicyos bulbosus

Eleven oleanane-type saponins (1-11) have been isolated from Microsechium helleri and Sicyos bulbosus roots and were evaluated for their antifeedant, nematicidal and phytotoxic activities. Saponins {3-O-β-d-glucopyranosyl (1 → 3)-β-d-glucopyranosyl-2β,3β,16α,23-tetrahydroxyolean-12-en-28-oic acid 28-O-α-l-rhamnopyranosyl-(1 → 3)-β-d-xylopyranosyl-(1 → 4)-[β-d-xylopyranosyl-(1 → 3)]-α-l-rhamnopyranosyl-(1 → 2)-α-l-arabinopyranoside} (1), and {3-O-β-d-glucopyranosyl-2β,3β,16α,23-tetrahydroxyolean-12-en-28-oic acid 28-O-α-l-rhamnopyranosyl-(1 → 3)-β-d-xylopyranosyl-(1 → 4)-[β-d-xylopyranosyl-(1 → 3)]-α-l-rhamnopyranosyl-(1 → 2)-α-l-arabinopyranoside} (2) were also isolated from M. helleri roots together with the two known compounds 3 and 4. Seven known structurally related saponins (5-11) were isolated from S. bulbosus roots. The structures of these compounds were established as bayogenin and polygalacic glycosides using one- and two-dimensional NMR spectroscopy and mass spectrometry. Compounds 7, 10, bayogenin (12) and polygalacic acid (13) showed significant (p < 0.05) postingestive effects on Spodoptera littoralis larvae, compounds 5-11 and 12 showed variable nematicidal effects on Meloydogyne javanica and all tested saponins had variable phytotoxic effects on several plant species (Lycopersicum esculentum, Lolium perenne and Lactuca sativa). These are promising results in the search for natural pesticides from the Cucurbitaceae family.     

Cloning and characterization of an extracellular serine protease from the nematode-trapping fungus <Emphasis Type="Italic">Arthrobotrys conoides</Emphasis>

An extracellular serine protease (Ac1) with a molecular mass of 35 kDa was purified from the nematode-trapping fungus Arthrobotrys conoides. The optimum activity of Ac1 is at pH 7.0 and 53.2°C (over 20 min). Ac1 can degrade a broad range of substrates including casein, gelatin, bovine serum albumin, collagen, and nematode cuticles. Moreover, the enzyme can immobilize the free-living nematode Panagrellus redivivus and the pine wood nematode Bursaphelenchus xylophilus, indicating Ac1 may be involved in infection against nematodes. The encoding gene of Ac1 contains one intron of 60-bp and two exons encoding a polypeptide of 411 amino acid residues. The deduced polypeptide sequence of Ac1 showed a high degree of similarity to two previously reported serine proteases PII and Mlx from other nematode-trapping fungi (81% aa sequence identity). However, three proteases Ac1, Aoz1 and Mlx showed optimum temperatures at 53.2, 45 and 65°C, respectively. Compared to PII, Ac1 appears to have a significantly higher activity against gelatin, bovine serum albumin, and non-denatured collagen. Moreover, our bioassay experiments showed that Ac1 is more effective at immobilizing P. redivivus than B. xylophilus.