水稻干尖线虫(Aphelenchoides besseyi)人工培养研究

利用真菌培养水稻干尖线虫的结果表明：不同的真菌培养线虫的效果有较大差异。链格孢菌（Alternaria sp.）、镰刀菌（Fusarium sp.）的效果明显优于常用的灰葡萄孢菌（Botrytis cinerea）,可选用于培养水稻干尖线虫。在真菌菌落上,不同的接虫量以及不同的温度和培养时间对水稻干尖线虫的培养均有一定的影响。     

水稻干尖线虫生殖方式、生活史及温度对其存活能力的影响及其在水稻中的分布

为了解近年我国水稻主产区水稻干尖线虫Aphelenchoides besseyi种群的生物学特性，并为寻找新的水稻干尖线虫病害防控方法，通过在胡萝卜愈伤组织、灰葡萄孢菌Botrytis cinerea和水稻中培养水稻干尖线虫的方法对其生殖方式、生活史和温度对其存活能力的影响及其在水稻中的分布进行研究。结果表明，在供试的12个水稻干尖线虫种群中，只有G8-315种群可孤雌生殖；在25℃下，水稻干尖线虫在胡萝卜愈伤组织和灰葡萄孢菌上生活史均约为12 d，在水稻苗上生活史约为11 d;-20℃以下低温或42℃以上高温处理12 h，在清水、胡萝卜愈伤组织和灰葡萄孢菌中水稻干尖线虫存活率均显著下降，处理72 h水稻干尖线虫存活率均为0；带虫干燥谷粒在-80、-20和4℃低温保存6个月后，谷粒内水稻干尖线虫存活率均显著低于25℃（对照），其中-80℃保存的谷粒内水稻干尖线虫存活率为0;56、70和80℃高温处理12 h后，2个水稻品种谷粒内水稻干尖线虫存活率均显著下降，80℃处理16 h后2个水稻品种谷粒内水稻干尖线虫存活率均为0；水稻幼苗各部位均可检测出水稻干尖线虫，其中假茎部位水稻干尖线虫数量...     

辽宁省大型真菌浸提液及发酵液对线虫的毒力作用

利用从辽宁省采集的206株大型野生真菌的浸提液和部分发酵液,测定其对根结线虫Meloidogyne incognita、胞囊线虫Heterodera glycines、小杆线虫Caenorhabditis sp.和水稻干尖线虫Aphelenchoides basseyi的活性。结果表明：有22种真菌对靶标线虫的致死率达70%以上,且同一菌株的浸提液和发酵液对线虫亦有明显的选择毒性作用。有7株菌浸提液对2种以上靶标线虫的致死率超过90%。蛇头菌Mutinus caninus F149浸提液对南方根结线虫毒力最高,致死率达97.74%,对胞囊线虫和水稻干尖线虫的毒力也超过90%,而对小杆线虫毒力仅为24.04%,因其对人毒性大而未获得纯培养。薄蜂窝菌Hexagonia tenuisF128浸提液对大豆胞囊线虫J2毒力最高,致死率达96.55%,但其发酵液毒力下降;浸提液对根结线虫的毒力仅次于F149,对水稻干尖线虫毒力较弱,但发酵液对干尖线虫致死率却提高至91.38%。块鳞青鹅膏Amanita excelsaF155浸提液对根结线虫的致死率达96.67%,但发酵液仅为70.93%。粉迷孔菌Daedalea biennis F139的浸提液对小杆线虫的毒力最强,致死率达90.40%。松生拟层孔菌Fomitopsis pinicola F130的浸提液与裂褶菌Schizophyllum commune F107的浸提液和发酵液及小灰孢Lycoperdon pusillus F102的发酵液对水稻干尖线虫的致死率均超过95%。血红菇Russula sanguinea F204和大红菇R.alutacea F153的浸提液和发酵液对3种植物线虫的致死率均超过85%,而对小杆线虫的毒力较弱。     

食线虫真菌蠕虫埃斯特菌对植物寄生线虫的侵染特性

为评价食线虫真菌蠕虫埃斯特菌Esteya vermicola菌株NKF 13222的生防应用价值,采用2%水琼脂培养基平板接种法测定了该菌株对7种植物寄生线虫的诱引、黏附和侵染能力,并比较了其对不同线虫的侵染特性.结果显示,该菌株的菌丝对松材线虫、拟松材线虫、腐烂茎线虫和水稻干尖线虫均有诱引作用,而对南方根结线虫、禾谷孢囊线虫和穿刺短体线虫无诱引作用;菌株的粘性分生孢子对有诱引现象的4种线虫都能黏附,接种24 h后的黏附率达到82%～99%,对无诱引现象的3种线虫不能黏附;粘性分生孢子黏附虫体后对不同线虫的侵染力存在差异,接种7 d后的松材线虫死亡率可达98%,而拟松材线虫、腐烂茎线虫和水稻干尖线虫死亡率分别为43%、35%和9%.表明蠕虫埃斯特菌不仅是松材线虫的强内寄生真菌,而且对其它重要植物寄生线虫也具有一定的应用潜力.     

花生茎线虫 Ditylenchus arachis 在真菌上的培养条件研究

花生茎线虫Ditylenchus arachis是危害花生的重要病原线虫。本研究通过测试不同真菌和培养温度对花生茎线虫繁殖力的影响,以探讨该线虫在室内的培养条件。研究结果表明:花生茎线虫在长柄链格孢、茄镰孢、尖镰孢和灰葡萄孢上均能完成生活史并进行繁殖,但不同真菌上培养线虫的效果不同。其中长柄链格孢的培养效果最好,培养30d后的总虫量和繁殖系数分别为37 292条和1 243倍;灰葡萄孢的培养效果最差,30d后的总虫量和繁殖系数仅为1 514条和50倍。25~30℃是花生茎线虫的适宜培养温度,30℃时为最佳温度;当温度低于20℃或高于35℃时线虫繁殖效果差。此方法繁殖的线虫可用于花生茎线虫生物学、遗传学等方面的研究。     

河北廊坊大豆枯萎病病原镰刀菌的分子鉴定

 为明确河北廊坊中国农科院植保所试验基地大豆孢囊线虫病田内大豆枯萎病病原镰刀菌的种类,对362份罹病枯萎大豆植株进行病原真菌分离,得到335株真菌;使用镰刀菌通用引物鉴定出镰刀菌(Fusarium spp.) 279株,占分离菌株83.3%;镰刀菌特异性引物、测序等分子生物学技术结合形态学特征进一步鉴定镰刀菌种类,鉴定出尖孢镰刀菌(F. oxysporum)189株,占分离菌株56.4%;茄病镰刀菌(F. solani)67株占20.0%、禾谷镰刀菌(F. graminearum)16株占4.8%、木贼镰刀菌(F. equiseti)3株、层出镰刀菌(F. proliferaum)2株、燕麦镰刀菌(F. avenaceum)和厚孢镰刀菌(F. chlamydosporum)各1株;致病性测试结果表明数量最多的尖孢镰刀菌(F. oxysporum)中约92.8%菌株具有不同程度的致病力;这些结果表明该试验基地大豆枯萎病的优势病原菌为尖孢镰刀菌(F. oxysporum);研究结果可为大豆枯萎病的防治提供科学依据,并为大豆孢囊线虫与尖孢镰刀菌复合侵染大豆的研究奠定基础。     

水稻干尖线虫室内人工培养条件研究

在室内同一培养基质上研究了温度、光照、湿度对水稻干尖线虫繁殖量的影响。结果表明:在不同温度、光照、湿度下水稻干尖线虫的繁殖量差异明显,在温度30℃,光照6h,湿度90%条件下水稻干尖线虫的繁殖量最高。     

快菜种传真菌检测和优势菌群的初步鉴定

针对北京地区主要快菜品种,利用PDA培养基培养自然干种子、无菌水洗涤处理的种子和1%NaClO处理的种子,开展带菌情况检测,并通过观察显微形态对优势菌群进行了初步鉴定。结果表明,10个快菜品种干种子总体携带的主要菌群依次为链格孢属(Alternaria spp.)、青霉属(Penicillium spp.)、曲霉属(Aspergillus spp.)、镰刀属(Fusarium spp.)真菌;无菌水洗涤后的种子及悬浮液中的优势菌群依次为链格孢属、青霉属、曲霉属真菌;1%NaClO处理的快菜种子的优势菌群为链格孢属真菌。其中,链格孢菌为快菜种传主要病原真菌。     

猕猴桃储藏期病原真菌的分离与鉴定

为明确猕猴桃采后病害的病原种类,本研究对猕猴桃储藏期病果进行了病原真菌的分离培养,观察了病原的培养性状和形态特征,并进行了分子生物学鉴定及其致病性测定。结果表明,引起猕猴桃储藏期病害的病原真菌有7种(类),分别是链格孢菌(Alternaria spp.)、葡萄座腔菌(Botryosphaeria dothidea)、间座壳菌(Diaporthe spp.),无性态为拟茎点霉菌(Phomopsis spp.)、拟盘多毛孢菌(Pestalotiopsis sp.)、假尾孢菌(Pseudocercospora sp.)、刺盘孢(Colletotrichum sp.)、灰葡萄孢(Botrytis cinerea),研究结果可为后期猕猴桃采后病害的防控提供科学依据。     

水稻干尖线虫快速分子检测技术研究

根据水稻干尖线虫的rDNA-ITS序列,设计出水稻干尖线虫的特异性引物,利用PCR技术对水稻干尖线虫(Aphelenchoides besseyi)部分rDNA-ITS1和部分5.8S基因核苷酸序列进行特异性扩增。实现了单条活的或4%的甲醛(FG)固定的水稻干尖线虫的快速检测。     

真菌寄生大豆孢囊线虫的初步研究

从山东省高密县、胶州市及江苏省沛县5个采集点的大豆孢囊线虫中,共分离出110个真菌菌株。优势真菌是镰刀菌,其它为柱孢菌、弯孢霉菌和一种子囊菌。真菌的出现机率平均为44％。在洋菜平板上测定,菌株对该线虫卵,均有不同程度的重寄生能力,其中茄病镰刀菌高达49％。该菌和柱孢菌的分生孢子萌发后可直接穿透卵壁,卵的内含物完全被其粗短的菌丝体取代。     

Influence of a Fungus-Feeding Nematode on Growth and Biocontrol Efficacy of Trichoderma harzianum

ABSTRACT A fungivorous nematode, Aphelenchoides sp., was isolated from field soil by baiting with mycelium of the biocontrol fungus Trichoderma harzianum ThzID1, and subsequently was maintained on agar cultures of the fungus. Interactions between the nematode and the green fluorescent protein-producing transformant, T. harzianum ThzID1-M3, were investigated in both heat-treated (80 degrees C, 30 min) and untreated field soil. ThzID1-M3 was identified in soil by epifluorescence microscopy. When ThzID1-M3 was added to soil as an alginate pellet formulation, addition of the nematode (10 per gram of soil) significantly reduced radial growth and recoverable populations of the fungus, and the effect was greater in heat-treated soil than in untreated soil. Addition of ThzID1-M3 to soil pretreated with the nematode (10 per gram of soil) stimulated nematode population growth for approximately 10 to 20 days, whereas nematode populations decreased in the absence of added Trichoderma sp. When sclerotia of Sclerotinia sclerotiorum were added to soil (10 per 200 g of soil) with ThzID1-M3 (40 pellets per 200 g of soil), addition of Aphe-lenchoides sp. (2,000 per 200 g of soil) reduced the number of sclerotia colonized by ThzID1-M3. These results suggest that fungivorous nematodes may be a significant biotic constraint on activity of biocontrol fungi in the field.     

根际微生物多样性与棉花品种对黄萎病抗性关系研究——Ⅰ.根际微生物数量与棉花品种对黄萎病抗性的关系

 对抗、感黄萎病6个棉花品种的根际线虫、真菌、细菌和放线菌的数量在苗期和现蕾期采用选择性培养基和直接观察法进行了分析。分析结果表明:棉花对黄萎病的抗性与根际真菌和放线菌数量呈正相关;与根际线虫数量负相关;与根际细菌数量无显著相关性。     

AM真菌和根结线虫互作对黄瓜生长及生理特征的影响

通过盆栽试验研究了丛枝菌根真菌Arbuscularmycorrhiza（AM）和南方根结线虫Meloidogyneincognita互作对黄瓜幼苗生长及生理特征的影响。结果表明,预先接种地表球囊霉Glomusversi—forme再接种线虫处理的鲜、干重较只接种线虫的处理显著增加;预先接种AM真菌再接种线虫处理的根结线虫病情指数、AM真菌侵染率较只接种线虫的处理明显降低;接种线虫使黄瓜植株的过氧化物酶（POD）、过氧化氢酶（CAT）及苯丙氨酸解氨酶（PAL）、总酚含量有不同程度提高,其中预先接种根内球囊霉G．intraradices对黄瓜植株起到了较好的保护作用;与对照相比,只接种线虫处理的叶片磷含量降低了25．26％,预先接种AM真菌再接种线虫处理的叶片磷含量较只接种根结线虫的含量高。     

根际微生物多样性与棉花品种对黄萎病抗性关系研究——Ⅰ.根际微生物数量与棉花品种对黄萎病抗性的关系

 对抗、感黄萎病6个棉花品种的根际线虫、真菌、细菌和放线菌的数量在苗期和现蕾期采用选择性培养基和直接观察法进行了分析。分析结果表明:棉花对黄萎病的抗性与根际真菌和放线菌数量呈正相关;与根际线虫数量负相关;与根际细菌数量无显著相关性。     

Effects of root- and stolon-infecting fungi on root-colonizing nematodes of white clover

The effects on white clover ( Trifolium repens ) of different combinations of the nematodes Meloidogyne trifoliophila , Helicotylenchus dihystera and Heterodera trifolii and nine stolon-infecting and three root-infecting fungi were studied in a glasshouse experiment. The presence of the fungus Phytophthora megasperma alone increased ( P  < 0·001) root-rot severity and reduced ( P  < 0·001) plant growth. Other species combinations, such as Phoma nebulosa and Alternaria alternata , interacted and increased root-rot severity. Combinations of P. megasperma with Pythium irregulare , and P. nebulosa with Phoma medicaginis or A. alternata , increased M. trifoliophila populations. Several other fungi ( P. irregulare , P. nebulosa , Colletotrichum coccodes , Macrophomina phaseolina , P. medicaginis and Phoma sp.) interacted with the nematode M. trifoliophila causing severe root-knot symptoms. The results indicated that fungi and nematodes interacted to cause root and stolon rot and reduced yields, and that poor persistence of white clover in pastures is likely to be a problem with a complex etiology.     

一株茶树内生淡紫拟青霉的鉴定及抗真菌活性研究

采用形态学观察及ITS序列分析对1株分离自健康茶树叶片的内生真菌进行鉴定,初步确定该菌株为淡紫拟青霉;并通过对峙培养和发酵液的抑菌试验研究其对真菌的抗生作用.结果表明,淡紫拟青霉FHM-1对7种茶树内生真菌和10种植物病原菌均有较强的抑菌作用;其发酵液对供试的16种真菌具有不同程度的抑制作用,而对拟盘多毛孢的生长有较强的促进作用.     

Suppression of Meloidogyne hapla by arbuscular mycorrhiza fungi (AMF) on pyrethrum in Kenya

Two arbuscular mycorrhiza fungi (AMF) isolated from a pyrethrum-growing region in Kenya were screened for efficacy against a nematode, Meloidogyne hapla, in greenhouses. The fungi were identified at INVAM (International Culture Collection of Vesicular Arbuscular Mycorrhizal Fungi) as Glomus etunicatum (Isolate KS18) and Glomus sp. (Isolate KS14). Isolate KS14 (Glomus sp.) significantly suppressed nematode population, growth and development by up to 54%, egg production by up to 75% and disease severity by up to 71%. Glomus etunicatum (Isolate KS18) suppressed nematode growth and development by up to 50%, egg production by up to 75% and disease severity by up to 57%. In addition, G. etunicatum and Isolate KS14, significantly improved top dry biomass of pyrethrum by up to 33% and 47%, respectively. Glomus etunicatum and M. hapla were mutually inhibitory as root colonization by G. etunicatum was significantly reduced (up to 24%) by the presence of the nematode. The presence of the nematodes, on the other hand, did not significantly affect root colonization by Isolate KS14.     

Population variation in traits of Deladenus siricidicola that could influence the biocontrol of Sirex noctilio in South Africa

Deladenus siricidicola is the primary biocontrol agent of the invasive woodwasp, Sirex noctilio. The nematode is mass-reared in culture on the S. noctilio symbiotic fungus, A. areolatum, but can also be induced to convert into its infective form in culture. The aim of this study was to explore the level of variation among a collection of D. siricidicola strains to convert into the infective form in culture. In addition, we measured variation in growth of the nematode on a strain of A. areolatum that is commonly used for laboratory rearing. Both traits could influence laboratory rearing and efficiency in the field. The 22 strains from three countries showed significant variation in the frequency of conversion into the infective form in culture. Even genetically uniform strains from South Africa displayed a range of variation in this trait. There was also significant variation in the growth of D. siricidicola on A. areolatum. These results emphasise the importance of considering variation in traits in D. siricidicola that could influence a biological control program. They also provide tools that could be used to screen larger collections of D. siricidicola for variation in traits linked to virulence and reproduction.