马铃薯疮痂病菌致病相关基因的克隆及表达

 A pathogenic-related gene nec1 was cloned in Streptomyces scabies CPS-1, potato scab strain. Analysis results showed that the length of open reading frame(ORF) for nec1 gene was 666 bp, and the GC content was 54.2%. Sequence alignment indicated that a 650 bp up-stream sequence shared 91% similarity with IS 256 family transposase nucleotide sequences by BLASTn searches against GenBank. The segments obtained by PCR amplification were digested by enzymes SphⅠand SacⅠ, and linked to the expression vector pIJ702. The recombinants were transformed into nonpathogenicity strain Streptomyces lividans 66 TK24. Bioas-say results suggested that the transformants possessed the same symptoms as pathogenic strain on potato tuber slices and radish seedlings, which implied that nec1 gene was associated with the pathogenicity of S. scabies CPS-1.     

蝴蝶兰软腐病中一种新致病菌的分离与鉴定

 Soft rot disease often affects Phalaenopsis amabilis during the growing season. However, the pathogen of the disease is remaining poorly studied. In this study, bacterial strain R1 was isolated from soft rot tissues in Wuhan. The pathogenic, morphological, physiological, biochemical tests and 16S rDNA sequence analysis were carried out. The homology of 16S rDNA sequence between strain R1 and Pseudomonas grimontii was 99.72%, and its physiological and biochemical properties were also similar to those of Pseudomonas grimontiis. All these evidences indicated that strain R1 could be identified as a novel strain of Pseudomonas grimontii. The pathogenicity of the novel isolate was proved according to the Koch's postulates. This is the first report that Pseudomonas grimontii can cause soft rot disease of Phalaenopsis amabilis.     

水稻条纹病毒NS2基因原核表达产物多克隆抗体制备及受侵染水稻和灰飞虱体内NS2检测

 The NS2 gene of Rice stripe virus (RSV) was amplified by RT-PCR, cloned into pGEM-T vector and sequenced. The NS2 gene was inserted into prokaryotic expression vector pET32a to produce recombinant plasmid pET32a-NS2. The recombinant plasmid was introduced into Escherichia coli strain BL21 (DE3) pLysS. SDS-PAGE and Western blot analysis confirmed that NS2 fusion protein was expressed after induction by IPTG. The recombinant NS2 protein was purified with Ni²⁺-NTA agarose affinity chromatography and the polyclonal antibody against NS2 protein was raised in rabbit. NS2 protein was successfully detected in small brown planthopper (Laodelphax striatellus) at 1:1 600 dilution of the total protein of single planthopper and in infected rice (Oryza sativa) at 1:800 dilution of 10 mg leave by dot immunobinding assay using the polyclonal antibody.     

玉米大斑病菌孢子萌发和附着胞形成的影响因素研究

 Factors of influence on conidium germination and appressorium formation of Setosphaeria turcica,such as light condition,conidial concentration,nutrient resources and pH value of conidial suspension were studied.There was no significant difference among light treatments.The optimal pH for conidium germination and appressorium formation was 5.0 to 7.0.The exogenous nutrient sources were not the indispensable factors for conidium germination,but 5% sugar solution were more favorable for appressorium formation than the others.Low conidial concentration in suspension (conidia ≤ 10⁴/mL) was propitious to conidium germination and appressorium formation,which were inhibited significantly in higher concentration.It was suggested that the phenomenon was due to the self-inhibitor,a kind of lipophilic substance,existing in the site of conidium germination.     

小麦抗白粉病基因Pm6的微卫星标记鉴定

 Wheat powdery mildew, caused by Blumeria graminis f. sp. tritici, is a prevalent disease worldwide. Breeding and planting resistance cultivars have been proved effective and environmental friendly for control of the disease. To develop easily used PCR-based markers in marker assisted selection (MAS) for Pm6, a dominant powdery mildew resistance gene in wheat, 25 microsatellite markers on chromosome 2BL in wheat were screened between susceptible parent Yumai13 and resistance parent Timgalen carrying Pm6. F₂ population derived from Yumai13 and Timgalen was further analyzed by the marker Xgwm501. The results indicated that Xgwm501 was a co-dominant marker linked to Pm6 gene at a distance of 14.8 cM. 29 Pm-carrying varieties were tested by the marker Xgwm501 and only those carrying Pm6 showed 117 bp resistance specific band. This marker is proved to have high practicability and can be used in MAS of Pm6 gene in wheat breeding programs.     

Clonostachys rosea 67-1液生孢子形成及其抗性研究

 The growth and sporulation of Clonostachys rosea strain 67-1 in PD broth was observed and figured out. A large amount of submerged spores were obtained in shake flask with proprietary Czapek me-dium. Meanwhile, the colonies of strain 67-1were cultured in PDA plate and aerial spores were collected by elution. Resistances of submerged spores and aerial spores to high temperature, dry condition and UV treatment were determined. The results showed that the survival of the submerged spores kept in 60℃ for 30 min was 76.7%, while the aerial spores were hardly germinated in the same condition. After two weeks' drying treatment, the spore activities were 89.2% and 29.3%, respectively. Similarly, the activity of submerged spores was 72.6% and that of aerial spores was 19.7% when exposing to UV for 1 min. It is illuminated that deep submerged fermentation is more efficient than solid culture for strain 67-1 to produce chlamydospores, which act as main component in biopesticide mass production.     

一些植物提取物的抗菌活性筛选

 Methanol-extracts from nine plant species were screened for their bioactivity against Peronophythora litchii, Xanthomonas oryzae pv. oryzae and Ralstonia solanacearum. The plant extracts were applied on detached litchi young leaves before inoculation with sporangial suspension of P. litchii. The results showed that methanol extracts (at conc. of 100mg dry methanol extract weight per mL) of Hamamelis mollis and Derris scabricanlis possessed the highest antifungal activity against litchi downy blight with the inhibitory effectiveness of 88.89%, followed by Phryma leptostachya (77.78%), Clematis armcmdii (66.67%), and Daphniphyllum macropodum (61.11%). At the dose of lmg dry methanol extract weight, none of the tested plant methanol extracts showed inhibitory activities against X. oryzae pv. oryzae, whereas, H. mollis had the strongest antibacterial activity against R. solanacearum with the inhibitory zone diameter of 21 mm, followed by Aristolochia mollissima, D. macropodum, Pachyrhizus erosus, Vicia unijuga, and C. armcmdii with the inhibitory zone diameter between 9 to 11 mm.     

菌寄生真菌纤细齿梗孢一丝氨酸-苏氨酸蛋白激酶基因OPK1的克隆及序列分析

 Using degenerate PCR and TAIL-PCR, a protein kinase gene OPK1 (Genebank accession No.:EU417815) was cloned from mycoparasite fungi Olpitrichum tenellum. OPK1 has an open reading frame of 2 031 bp interrupted by two introns (108 bp, 84 bp) and putatively encodes a protein of 676 aa. Phylogenetic analysis indicated that OPK1 was most similar to other serine-threonine protein kinase in fungi. Southern blot analysis indicated that OPK1 is present as a single copy in the genome. RT-PCR showed it could be transcribed both in the phase of spore germination and hyphal growth.     

Nematicidal activity of anion transport blockers against Meloidogyne incognita, Caenorhabditis elegans and Heterorhabditis bacteriophora

BACKGROUND: Because methyl bromide has been phased out as a soil sterilant, new nematicides are urgently needed. Four different chemical classes of organic acids acting as anion transport (AT) blockers were tested against a free-living nematode, Caenorhabditis elegans Maupas, a plant-parasitic nematode, Meloidogyne incognita (Kofoid and White) Chitwood, and an entomopathogenic nematode, Heterorhabditis bacteriophora Poinar, in toxicity bioassays. The materials tested were DIDS (4,4'-diisothiocyanatostilbene-2,2'-disulfonic acid), 9-AC (anthracene-9-carboxylic acid), NPPB [5-nitro-2-(3-phenylpropylamino)benzoic acid] and IAA-94 (indanyloxyacetic acid). RESULTS: All the compounds showed slowly developing nematicidal activity against second-stage juveniles of M. incognita and adults of C. elegans, but not against H. bacteriophora infective-stage juveniles. The LC(50) values of these compounds were < 50 mg L(-1) after 48 and 72 h incubation, while at 168 h incubation the LC(50) values were < 10 mg L(-1) for both sensitive species. Across both species and time, the LC(50) values generally differed no more than twofold among the four compounds tested in this study. In contrast, none of the compounds (200 mg L(-1)) caused more than control mortality to H. bacteriophora, even after 168 h of incubation. CONCLUSION: These compounds are potential leads for commercial nematicides. The insensitivity to H. bacteriophora is consistent with the natural exposure of this nematode to DST (3,5-dihydroxy-4-isopropylstilbene), a stilbene produced by its symbiotic bacterium. Based on the known activity of the compounds used in this study, it is suggested that anion transporters form the probable target sites for DIDS, 9-AC, NPPB and IAA-94 in nematodes.     

4种杀虫剂对甘草胭珠蚧的防治效果

甘草胭珠蚧Porphyrophora sophorae(Arch.)是人工栽培甘草的主要害虫。采用灌根、土壤处理法研究了35%吡虫啉悬浮剂、70%噻虫嗪水分散粒剂、48%毒死蜱乳油和5%毒死蜱颗粒剂4种杀虫剂对甘草胭珠蚧的防治效果。结果表明,8月中旬施药各药剂的防治效果均低于50%,但可有效降低翌年春季田间的虫口密度;6月中旬采用35%吡虫啉悬浮剂和70%噻虫嗪水分散粒剂灌根的防效可达80%以上。建议生产中可在6月中旬或之前采用灌溉后进行35%吡虫啉悬浮剂1 500倍或70%噻虫嗪水分散粒剂7 500倍液300mL/株灌根的措施防治甘草胭珠蚧。     

3种环境友好型药剂对西花蓟马的室内毒力与田间防效

测定了3种杀虫剂对西花蓟马的室内毒力和田间防效。室内生物测定结果表明,多杀菌素、甲氨基阿维菌素苯甲酸盐、阿维菌素对西花蓟马成虫的LC50值为0.050～2.887mg/L,对西花蓟马若虫的LC50值为0.040～0.457mg/L;田间药效试验表明,田间防治西花蓟马时推荐使用剂量(防效>80%)分别为:2.5%多杀菌素悬浮剂30～60g/667m2,1%甲氨基阿维菌素苯甲酸盐乳油30～60g/667m2,1.8%阿维菌素乳油不可低于75g/667m2。     

Cross-resistance potential of fipronil in Musca domestica

The toxicity of fipronil to insecticide-susceptible houseflies and the cross-resistance potential of fipronil were determined for six insecticide-resistant laboratory housefly strains by topical application and feeding bioassay. The insecticide-resistant strains represented different levels and patterns of resistance to pyrethroids, organophosphates, carbamates and organochlorines. Five strains were almost susceptible to fipronil in feeding bioassay with resistance factors at LC50 between 0.36 and 3.0. Four of these strains were almost susceptible to topically applied fipronil (resistance factors at LD50 were 0.55, 0.83, 3.3 and 2.5, respectively), whereas one strain was 13-fold resistant to topically applied fipronil. A highly gamma-HCH-resistant strain, 17e, was 430-fold resistant to fipronil in topical application bioassay and 23-fold resistant in feeding bioassay at LD50/LC50. We also tested the toxicity of fipronil in a feeding bioassay and gamma-HCH in topical application bioassay on thirteen housefly field populations. Eleven of the field populations had resistance factors for fipronil ranging from 0.98 to 2.4 at LC50, whereas two populations were 4.0- and 4.6-fold resistant to fipronil. The resistance level to gamma-HCH at LD50 in the field populations ranged from 1.8- to 8.1-fold. The two strains showing fipronil resistance were 3.4- and 8.1-fold resistant to gamma-HCH. Fipronil and gamma-HCH toxicities were positively correlated in the field populations. Biochemical assays of esterase, glutathione S-transferase and cytochrome P450 monooxygenase indicated that the low fipronil resistance observed in laboratory and field strains could be caused by elevated detoxification or be due to a target-site resistance mechanism with cross-resistance to gamma-HCH.     

苏云金芽孢杆菌对松材线虫的杀线活性

在室内条件下,测定了17株苏云金芽孢杆菌Bacillus thuringiensis伴孢晶体蛋白对松材线虫Bursaphelenchus xylophilus的杀线活性。通过初筛,发现菌株020、RBT-200701的伴孢晶体蛋白对松材线虫有明显毒杀作用。在测试的浓度范围内（菌株020为6.37-203.9μg/ml;RBT-200701为13.7-438.8μg/ml）,菌株对松材线虫的校正死亡率随其浓度增大而提高;随时间延长杀虫效果提高。020菌株晶体蛋白24h、48h时对松材线虫的LC50、LC95分别为183.20、408.20μg/ml和80.132、25.29μg/ml;RBT-200701菌株伴孢晶体蛋白24、48h时对松材线虫的LC50、LC95分别为368.00、868.00μg/ml和82.73、382.73μg/ml。SDS-PAGE分析发现这2个菌株晶体蛋白的蛋白图谱不同。     

海洋放线菌M1D14代谢产物对几种重要植物寄生线虫的抑制作用

采用梯度稀释法室内测定了海洋放线菌M1D14发酵液代谢产物对南方根结线虫(Meloidogyne incognita)2龄幼虫及其卵、松材线虫(Bursaphelenchus xylophilus)、大豆孢囊线虫(Heterodera glycines)2龄幼虫的毒力,同时还测定了经过高温高压灭菌和0.22μm微孔滤膜过滤的M1D14发酵液对南方根结线虫的毒力。结果表明,海洋放线菌M1D14发酵液代谢产物对南方根结线虫卵的孵化无明显的抑制作用,对南方根结线虫幼虫毒力最强,对松材线虫幼虫毒力次之,对大豆孢囊线虫幼虫的毒力最弱。经过高温高压灭菌和微孔滤膜过滤的M1D14发酵液对南方根结线虫仍有毒力。M1D14杀线虫谱广、效果好、使用成本低,在生产上具有很好的应用前景。     

对小菜蛾高毒的苏云金杆菌菌株筛选试验初报

从田间自然死亡的小菜蛾虫尸中分离到Ｂ．ｔ．９４１９茵株，经复壮选育及摇瓶考核，在自选培养基上生长良好，２４ｈ孢子开始脱落形成晶体，３６ｈ全部形成晶体。晶体大、宽棱形、角尖。４８ｈ发酵液含菌量为３５亿／ｍＬ。经发酵液对小菜蛾毒力测定，４８ｈＬＣ_５０：对照ＨＤ－１含菌量为２７４ｍＬ／Ｌ，而９４１９仅１２４ｍＬ／Ｌ。     

日光温室防治棕榈蓟马药剂筛选

采用浸叶法开展了7种杀虫剂对温室棕榈蓟马成虫的室内毒力测定,并对效果较好的2种药剂进行田间药效试验。室内生物测定结果表明,多杀菌素、甲氨基阿维菌素苯甲酸盐对棕榈蓟马成虫的LC50分别为0.079 mg/L 和0.443 mg/L,而阿维菌素、灭多威、啶虫脒、高效氯氟氰菊酯LC50值为47.908 ~206.236 mg/L。田间药效试验结果表明,2.5%多杀菌素悬浮剂和2.2%甲氨基阿维菌素苯甲酸盐微乳剂使用剂量以80~60 g/667 m2和 30~20 g/667 m2防治效果较理想。     

Susceptibility to thiamethoxam of Musca domestica from Danish livestock farms

BACKGROUND: Neonicotinoid baits are currently replacing anticholinesterase baits for control of adult houseflies (Musca domestica L.). Introduction of new insecticides includes evaluation of their cross-resistance potential, which was assessed for thiamethoxam in field populations from Denmark. RESULTS: In feeding bioassay with a susceptible strain, thiamethoxam LC(50) at 72 h was 1.7 microg thiamethoxam g(-1) sugar, making it 19-fold, 11-fold and threefold more toxic to houseflies than azamethiphos, methomyl and spinosad respectively. The field populations were 6-76-fold resistant to thiamethoxam. There was no correlation between the toxicities of thiamethoxam and spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The toxicity in feeding bioassay at 72 h of imidacloprid in a susceptible strain was 32 microg imidacloprid g(-1) sugar at LC(50), making it 19-fold less toxic to houseflies than thiamethoxam. There was a strong significant correlation between the toxicities of thiamethoxam and imidacloprid in field populations.CONCLUSION: Neonicotinoid-resistant houseflies were present at a detectable and noticeable level before thiamethoxam and imidacloprid were introduced for housefly control in Denmark. The toxicity of thiamethoxam is explained by other parameters than the toxicities of spinosad, dimethoate, methomyl, bioresmethrin or azamethiphos. The cross-resistance between thiamethoxam and imidacloprid indicates a coincidence of mechanism of the toxicity and resistance in the field populations.     

沼液对韭菜迟眼蕈蚊的防治效果

通过室内选择性测定和田间试验,评价了沼液对韭菜迟眼蕈蚊的防治效果。结果表明,沼液对韭菜迟眼蕈蚊表现出一定的驱避活性,对幼虫的驱避率、拒食率分别达55.30%、53.81%,雌雄成虫选择反应率分别为33.2%、30.5%,对雌虫产卵驱避率为36.82%。田间试验结果表明,以沼液∶水(1∶1)处理幼虫、成虫,随着时间的延长防效逐渐提高,处理后45d防效达到最大,分别为43.16%、56.50%。