Effect of antagonistic Fusarium spp. and of different commercial biofungicide formulations on Fusarium wilt of basil (Ocimum basilicum L.)

The ability of antagonistic Fusarium spp. to control Fusarium wilt of basil caused by F. oxysporum f. sp. basilici was tested under glasshouse conditions from 1994 to 1996. Fusarium oxysporum antagonistic strain 251/2, a protoplast fusion-derived hybrid coded FI-11 and F. moniliforme strains TF4 and TF4 RB were able to decrease (P = 0.05) the incidence of Fusarium wilt significantly. Biocontrol was consistent, especially when the antagonists were applied by mixing a talc preparation of propagules in the substrate (10⁵ CFU/ml of substrate) 1–2 weeks before sowing or at sowing. Seed coating with the biocontrol strains (10⁸ CFU/g of seed) did not generally provide sufficient disease containment, whereas the combination of substrate treatment and seed coating did not improve wilt control in comparison with substrate treatment alone. Two commercial formulations, based on antagonistic F. oxysporum strains and one containing the antagonistic strain K61 of Streptomyces griseoviridis, were generally not sufficiently effective at the tested dosages.     

韭菜根系分泌物对苦瓜枯萎病菌的化感作用

为探究韭菜根系分泌物对苦瓜枯萎病菌的化感作用,本研究以‘山东雪韭6号’韭菜和苦瓜枯萎病菌为试验材料,水培法收集韭菜根系分泌物,采用菌丝生长速率和孢子萌发法研究韭菜根系分泌物对苦瓜枯萎病菌的化感作用;利用气相色谱-质谱联用技术（GC-MS）鉴定根系分泌物组成成分,筛选可能的主效化感物质,进一步通过体外抑菌试验验证其化感作用,明确主效化感物质。结果表明,随着韭菜根系分泌物浓度的升高,苦瓜枯萎病菌的菌落直径和孢子萌发数均呈先降后升的趋势,当分泌物浓度为3.22 mg/mL时,抑制作用最大,满足化感作用低浓度促进、高浓度抑制的特性。通过GC-MS鉴定出韭菜根系分泌物中的主要成分为酚类、酯类、烷烃类、醇类、烯类、芳香类,可能的化感物质为邻苯二甲酸二丁酯、2,4-二叔丁基苯酚、2,2°-亚甲基双-(4-甲基-6-叔丁基苯酚)。通过体外抑菌试验证实这3种物质对苦瓜枯萎病菌菌丝生长和孢子萌发都有不同程度的化感抑制作用,且抑菌效果为2,4-二叔丁基苯酚>邻苯二甲酸二丁酯>2,2°-亚甲基双-(4-甲基-6-叔丁基苯酚),明确其为化感物质。由此可知,韭菜根系分泌的主效化感物质对苦瓜枯萎病菌有显著的化感抑制作用,这为苦瓜间套轮作韭菜防控苦瓜枯萎病和研制植物源抑菌剂提供了理论依据和技术支撑。     

In vitro assessments of diverse plant pathogenic fungi treated with a novel growth control agent

The efficacy of an agent with an iodine-based active ingredient (a.i.) was evaluated for controlling the growth of fungi pathogenic to many different food crops. Even though iodine is a necessary mineral for mammals and is an approved food additive, interest in using iodine-based agrochemicals for fungal control is recent. Fusarium verticillioides (synonym=F. moniliforme J. Sheld) sensitivity to the iodine-base agent was determined using two methods. One method used an agar plug taken from an actively growing culture and the other method used isolated conidia. The concentration of the agent required to inhibit growth of F. verticillioides was greater for the culture plug than isolated conidia. Forty-eight strains of F. verticillioides were analyzed representing different states within the United States, countries outside the United States, monocot and dicot plant hosts, and mating types. All strains of F. verticillioides, regardless of geographic origin, plant host, or mating type were sensitive to the test compound. In addition, 25 species of fungi pathogenic to a diverse array of crops were analyzed and demonstrated to be sensitive to this agent. A pesticide developed with an iodine-based a.i. could be an effective control for diverse fungi pathogenic to a range of plant hosts.     

与尖孢镰刀菌枯萎病相关的抑病型土壤研究进展

由尖孢镰刀菌引起的枯萎病是制约作物生产的世界性重要土传维管束病害,危害的作物种类多,损失严重,生产中尚无经济有效的防治方法。抑病型土壤是指对土传作物病害有抑制作用的一类土壤,具有有益微生物种群结构稳定、对土传病害防效持久等特点,因此国内外专家进行了许多卓有成效的研究。本文对尖孢镰刀菌枯萎病相关抑病型土壤的形成因素、作用机制等方面的研究进展进行综述,并对今后尖孢镰刀菌枯萎病抑病型土壤研究的重点方向及应用前景进行了分析和展望,以其为下一步深入研究尖孢镰刀菌枯萎病抑病型土壤的微生物种群结构、有益微生物种类构成、抑病型土壤的构建方法及复合微生物菌肥研制提供新的参考。     

Long-term dry preservation of viable mycelia of two mycoherbicidal organisms

The production of large amounts of fungal spores for preservation and formulation are considered constraints to effective use of fungal biocontrol agents. Few successful attempts have been made to store fungal mycelia alone. Late log-phase liquid fermenter cultured isolated mycelia of two Fusarium spp. specific to the parasitic broomrapes (Orobanche spp.) from fermenters were formulated in alginate beads or in ‘Stabileze’ (starch, sucrose, corn oil, and silica) and air-dried. ‘Stabileze’ formulations exhibited <30% loss of mycelial viability for >9 months and retained pathogenicity to the weed for over a year, while mycelia harvested earlier, and conidia from liquid culture exhibited >40% loss of viability. Mycelia from liquid culture yielded >20 times more colony forming units (cfu) of F. arthrosporioides and >2 times more cfu of F. oxysporum than spores at late log phase. Efficient formulation of mycelia should significantly change the economics of biocontrol.     

不同遗传背景香蕉品种间轮作控制香蕉枯萎病的效果分析

香蕉种质资源丰富,不同香蕉品种对各个枯萎病生理小种的耐受性不同,利用遗传背景差异大、抗性不同的香蕉品种进行轮作可能达到防控香蕉枯萎病的效果.本研究选取华南地区常见的香牙蕉、粉蕉、龙牙蕉、贡蕉等栽培蕉类型作为研究材料,通过对24个香蕉品种的遗传多样性进行分析,结合生产应用,筛选遗传背景不同并且具有一定抗枯萎病的品种,进行...     

香蕉枯萎病菌分子检测研究进展

香蕉枯萎病是全世界香蕉产业共同面临的毁灭性病害,但目前生产上仍缺乏适宜的抗病品种和有效的治疗措施。因此,借助快速准确的枯萎病菌检测技术及时明确病原菌以控制该病的传播和蔓延显得尤为重要。本文回顾了近年来国内外香蕉枯萎病菌分子检测技术的发展历程,归纳和总结了DNA指纹图谱、普通PCR、多重PCR、荧光定量PCR及等温扩增技术在该病菌检测中的研究进展,分析了不同检测技术的优缺点,并指出可能存在的问题和研究发展方向,为该病的分子检测技术优化和防控策略制定提供参考。     

海南广藿香内生真菌分离鉴定及拮抗菌株筛选

本研究从海南广藿香的根、茎、叶和花等组织中分离获得61株内生真菌,采用显微形态观察和ITS序列分析鉴定为15个属,其中优势菌群为炭疽菌属（Colletotrichum）和干酪酵母菌属（Meyerozyma）。以澳洲坚果叶枯病菌（Pestalotiopsis microspora）、西瓜枯萎病菌（Fusarium oxysporum）和火龙果溃疡病菌（Neoscytalidium dimidiatum）为指示菌,采用平板对峙法进行拮抗菌株筛选,发现镰刀菌属（Fusarium）菌株PfuJ20、PfuG16和PfuG5对澳洲坚果叶枯病菌的抑制效果最好,棒孢属（Corynespora）菌株PfuH2对西瓜枯萎病菌抑菌效果较好。采用滤纸片扩散法进行各菌株对金黄色葡萄球菌、大肠杆菌、沙门氏菌、枯草芽孢杆菌等动植物病原细菌的抑菌作用评价,发现镰刀菌属PfuJ20、PfuG16对各细菌均有较强抑制效果,而链格孢属（Alternaria） PfuH1对金黄色葡萄球菌的生长抑制效果相对最强。结果表明,海南广藿香内生真菌具有一定的生物多样性,部分内生真菌菌株具有较强的抑菌活性。     

豆科绿肥残体对连作蕉园土壤肥力的影响

采用室内恒温箱培养方法,探讨2种豆科绿肥残体对连作蕉园土壤理化性质和可培养微生物数量的影响。以连作障碍严重的蕉园土壤为研究背景,共设置4个处理：无残体添加（CK）、添加田菁残体（S）、添加白三叶残体（W）和添加香蕉残体（B）。结果表明：（1）在培养第5天时,与CK相比,B、S和W添加后均增加了土壤可培养尖孢镰刀菌、真菌、细菌的数量;然而在培养第45天,S和W尖孢镰刀菌数量逐渐下降。45 d时,与B相比,S和W尖孢镰刀菌数量分别降低33.09%和79.92%;（2）与CK和B相比,S显著降低土壤pH,而W能显著提高pH和速效钾含量;（3）主坐标分析 (PCoA) 和多元回归树分析 (MRT) 结果均说明,添加不同豆科绿肥残体（S和W）与CK差异显著,说明不同豆科绿肥残体添加可改善土壤肥力。因此,在高发病香蕉园地,间套作白三叶可改善其土壤理化和土壤可培养微生物状况,对香蕉连作障碍有较好的缓解作用。     

Application of an emulsifiable mixture of 1,3-dichloropropene and chloropicrin against root knot nematodes and soilborne fungi for greenhouse tomatoes in Italy

A mixture of 1,3-dicloropropene 60.5% w/w and chloropicrin 33.3% w/w (Telone C35 EC) may be registered in Italy for soil drip fumigation. Five experiments on greenhouse tomatoes in Northern, Central and Southern Italy compared the effectiveness of this mixture in comparison with methyl bromide to find the optimum application rate in soils infested by Fusarium oxysporum f.sp. lycopersici, F. oxysporum f.sp. radicis lycopersici, Sclerotium rolfsii, Meloidogyne javanica and M. incognita. Its efficacy against F. oxysporum f.sp. radicis lycopersici and M. incognita was confirmed when applied to soils at 100, 200, 300 and 400 l ha⁻¹ (132.4, 268.4, 402.6 and 536.8 kg ha⁻¹) under gas-tight films with 15–45 mm of application water (900–1200 mg Telone C35 EC l⁻¹). In sandy soils, with slight F. radicis lycopersici infections and with heavy nematode (M. incognita) attacks, the mixture, drip applied at 900 mg l⁻¹ during late summer (fumigation: late summer; transplant: late-summer/autumn; last harvest: early spring), performed well up to 132.4 kg ha⁻¹ (100 l ha⁻¹). In sandy loam soils with slight F. radicis lycopersici infections and severe infections of F. lycopersici and galling nematodes (M. javanica), 268.4 kg ha⁻¹ (200 l ha⁻¹) of the mixture applied at 900 mg l⁻¹ as a drip provided yields similar to those of methyl bromide treated plots both in spring and summer cycles. In sandy loam soils, the diseases (F. lycopersici, F. radicis lycopersici) were controlled at rates 268.4 kg ha⁻¹ (containing 90 kg ha⁻¹ of chloropicrin), but the mixture was ineffective against Sclerotium rolfsii occasionally observed in sandy loam soils. In both sandy and sandy loam soils, no significant relationships were found between the rates of mixture applied (132.4, 268.4, 402.6 and 536.8 kg ha⁻¹) and the degree of nematode infestation.     

香蕉枯萎病菌4号生理小种smy1基因的功能分析

Smy1是一种参与调控真菌生长的驱动蛋白。本研究克隆了香蕉枯萎病菌4号生理小种（Foc 4）驱动蛋白基因smy1,并采用同源重组技术获得smy1敲除的野生型Foc 4的smy1敲除突变体。通过测定突变体菌丝形态、生长速度、产生孢子量、对香蕉苗的致病力及对氰烯菌酯的敏感性,用于研究smy1在香蕉枯萎病菌的生长发育及对氰烯菌酯抗药性中的作用。结果表明,与野生型Foc 4相比,smy1敲除突变体生长缓慢、菌丝畸形、产孢量增加,对巴西蕉苗的致病力减弱,但对氰烯菌酯的抗性水平变化不大。由此推断smy1基因可能在Foc 4的生长发育、产孢以及致病力等方面具有重要作用。     

Efficacy of 1,3-dichloropropene plus chloropicrin application methods in combination with pebulate and napropamide in tomato

Small-plot research and large-field validations were conducted to determine the effect of 1,3-dichloropropene (1,3-D)+chloropicrin (Pic) application methods in combination with the herbicides pebulate and napropamide on pest control in fresh market tomato (Lycopersicon esculentum Mill.). In both the small-plot trials and the large-validation fields, various treatments compared the efficacy of soil fumigants and herbicides based on methyl bromide, in-bed and broadcast applications of 1,3-D+Pic, pebulate, and napropamide. The results consistently indicated that either in-bed or broadcast applications of 1,3-D+Pic in combination with pebulate and napropamide are equally effective against the weed Cyperus spp., the nematodes Tylenchorhynchus spp., Belonolaimus longicaudatus, and Meloidogyne spp., and the soilborne disease Fusarium oxysporum f.sp. lycopersici in fresh market tomato. Therefore, broadcast 1,3-D+Pic application can reduce personnel exposure and poisoning risks, without losing pest control efficacy.     

尖孢镰刀菌古巴专化型4号生理小种的磷酸化应激诱导蛋白1（FoSTIP1）基因的克隆及表达分析

本研究克隆了尖孢镰刀菌古巴专化型4号生理小种（Fusarium oxysporum f. sp. cubense race 4, Foc4）转录因子FoSkn7一个候选的下游基因,结果表明该基因cDNA编码序列全长1737 nt,编码一个含578个氨基酸残基的蛋白质。对其编码的蛋白进行了结构域和进化分析,结果表明该蛋白质含有胁迫诱导蛋白（stress-in-ducible protein-1, STI1）结构域和多个三角形4肽重复序列结构域（tetratricopeptide repeat, TPR）。该蛋白质与尖孢镰刀菌番茄专化型的磷酸化应激诱导蛋白1（STIP1）亲缘关系最近,因此初步将其确定为Foc4的磷酸化应激诱导蛋白并命名为FoSTIP1。采用相对荧光定量PCR方法分析了该基因在野生型B2菌株入侵香蕉苗根部及在外源H₂O₂诱导情况下的表达变化,也分析了FoSKN7基因缺失突变体中该基因在外源H₂O₂诱导情况下的表达。结果表明在入侵香蕉苗根部及在外源H₂O₂诱导情况下,B2菌株中的FoSTIP1表达均有上调,而FoSKN7基因缺失突变体中FoSTIP1即使有H₂O₂诱导,其表达也远低于B2菌株中的FoSTIP1。推测FoSTIP1可能是FoSkn7的靶基因并参与了Foc4抗外源氧化胁迫。     

三个小麦赤霉病抗源的抗性QTL定位

为寻找小麦赤霉病抗性基因及可用于分子标记辅助育种的抗性连锁标记．对中国的三个小麦赤霉病抗源苏麦3号、望水白和宁894037进行了抗性QTL的定位研究。SSR、AFLP分析与QTL分析结果表明,尽管三个抗源的来源和遗传背景并不同,但均在3B染色体短臂上发现抗性主效QTL,不同遗传群体所获得的QTL位点所处的染色体区段略有差异,位于QTL两翼的SSR标记也有所不同。苏麦3号的赤霉病抗性主效QTL位于3B染色体上的标记区间Xgwm533～Xgwm493内;宁894037的抗性位点分布于3B和6B染色体上。分别定位于标记Xgwm493～Xbarcl33和Xgwm644-Xgwm518之间;望水白的抗性主效QTL也位于3B染色体上．定位于标记Xgwm493～Xbarc147之间。微效QTL由于遗传群体的不同,分别住于1B、3B和2A染色体上。研究还表明,寻找抗性QTL在3B染色体以外的新抗源十分必要。     

甘蔗梢腐病菌Fusarium sacchari CYP51基因克隆及遗传转化

甾醇14α-去甲基化酶（CYP51）是生物细胞膜合成所需的一个非常重要的酶,在病原菌的耐药性、致病性和生长繁殖等方面发挥着非常重要的作用。研究表明干扰真菌的CYP51基因表达导致其无法正常生长,显著降低其致病性。本研究以甘蔗梢腐病菌甘蔗镰刀菌（Fusarium sacchari）为试验材料,根据基因组测序数据设计特异性引物克隆得到了FsCYP51基因全长和CDS全长。生物信息学分析表明,该基因序列全长1947 bp,编码区由2个内含子和3个外显子组成,CDS全长1554 bp,编码517个氨基酸,编码蛋白理论相对分子量为58.61 kDa。其编码蛋白的二级结构主要由α-螺旋和无规卷曲构成,具有典型的CYP51保守结构域。预测其亚细胞定位于细胞膜,且存在2个跨膜区域。系统进化分析表明,FsCYP51基因属于CYP51C这一类,与串珠镰刀菌（F. verticillioides）的CYP51C基因亲缘关系最近。同时,根据克隆到的基因全长和CDS全长构建了多价HIGS植物表达载体,并利用基因枪介导的遗传转化方法将干扰片段成功转化至甘蔗受体材料,为研究FsCYP51基因功能和创制抗梢腐病甘蔗种质奠定基础。     

香蕉枯萎病菌内源报告基因比卡菌素聚酮合酶编码基因Bik1的鉴定及应用

香蕉枯萎病是由尖孢镰刀菌古巴专化型(Fusarium oxysporum f.sp.cubense,Foc)引起的香蕉毁灭性土传病害,其中4号生理小种(Foc4)能感染几乎所有的香蕉品系,危害最严重.本研究克隆鉴定Foc4比卡菌素聚酮合酶编码基因(bikaverin PKS-encoding gene,Bik1)Foc...     

Preventing spread of Fusarium wilt of Hiemalis begonias in the greenhouse

To develop strategies to manage Fusarium wilt of Hiemalis begonias (Begonia×Hiemalis), caused by Fusarium foetens, studies were conducted to understand how the disease might be spread in the greenhouse. Inoculum density studies showed that as few as 100 conidial ml⁻¹ were sufficient to cause significant disease indicating that shared irrigation systems need strict sanitation along with bench tops, pots, and trays. The role of fungus gnats (Bradysia spp.) in vectoring F. foetens within a greenhouse was studied using nylon tent cages with diseased and healthy Hiemalis begonias. Fungus gnats were released into half of the cages. In cages where fungus gnats were present only healthy plants became diseased, indicating the effectiveness of fungus gnats in spreading Fusarium wilt. The pathogen was also isolated from adult fungus gnats. In vitro studies showed that two commercially available H₂O₂-based compounds, ZeroTol^® (2.0% peroxyacetic acid and 27.0% hydrogen peroxide), and SaniDate^® (12.0% peroxyacetic acid and 18.5% hydrogen peroxide), were effective in causing 100% spore mortality at rates that would allow their use in irrigation water. Seven cultivars of Hiemalis begonias (Begonia×Hiemalis) grown in soil infested with F. foetens were highly susceptible. No other Begonia species showed typical symptoms of chlorosis and wilt, but two cultivars of Rex begonia (Begonia rex) exhibited significant stunting in repeated trials.     

芽胞杆菌JK05的鉴定及其对香蕉、玉米的促生和生防潜能研究

解淀粉芽胞杆菌、枯草芽胞杆菌和苏云金芽胞杆菌等是重要的有益微生物,被广泛应用于植物病虫的生物防治。前期分离获得1株芽胞杆菌JK05,对其形态、生理生化特征、16S rRNA和gyrA基因序列进行分析,将其鉴定为解淀粉芽胞杆菌植生亚种Bacillus amyloliquefaciens subsp. plantarum。对峙培养实验结果显示,JK05菌株对多种植物病原镰刀菌具有拮抗作用。植物生长促进实验表明,JK05菌株对香蕉和玉米生长具有明显促进作用。盆栽实验结果显示,JK05菌株对香蕉枯萎病具有良好的防治效果。采用特异引物对JK05菌株基因组DNA进行PCR扩增,其可扩增到表面活性素（surfactin）、丰原素（fengycin）、伊枯草菌素A（iturin A）等抗生素合成基因。综上,JK05菌株具有良好的生防潜力,有望应用于生物农药和微生物肥料。     

Root diseases, weeds, and nematodes with poultry litter and conservation tillage in a sweet corn–snap bean double crop

The effect of continuous applications of poultry litter on root diseases, nematodes, and weeds with different tillage practices in vegetable production in Georgia was investigated. Litter treatments (noncomposted and composted broiler or breeder litter) applied 3–5 weeks before planting were compared with no litter. In sweet corn and snap bean there were differences among years, and tillage and litter treatments had a variable effect on yield, post-emergence damping-off and plant stand. Crown and brace root rot in corn induced by Rhizoctonia solani AG 2-2 increased after 3 years conservation tillage, but there were no differences among litter treatments. Population densities of R. solani AG-4 were increased by conservation tillage before planting snap bean the first year but not in later years, while litter treatments had no effect. After 3 years, populations of Pythium spp., Fusarium solani, and Fusarium spp. did not differ among treatments, but noncomposted broiler litter increased populations of saprophytic fungi in soil compared with no litter. Population densities of stylet-bearing nematodes were low and usually not different among treatments. After 3 years of sweet corn populations of Meloidogyne incognita increased with conventional tillage compared with conservation tillage, populations of Paratrichodorus christiei were reduced by litter treatments and populations of Helicotylenchus dihystera were reduced by noncomposted broiler litter compared with no litter. Weed infestation increased in both tillage systems with time, but in snap bean there was a greater infestation in conservation tillage than in conventional tillage. Litter treatments had little effect on weed infestation.     

中粒种咖啡新发砖红镰刀叶枯病病原菌鉴定及其病原生物学分析

针对引起海南省白沙农场中粒种咖啡成株叶片呈砖红色枯萎症状的一种未知病害的病原菌进行分离，分离得到菌株21BS02-1和21BS02-2。将分离菌株接种到叶片并观察症状，发现菌株21BS02-1的发病症状与田间症状一致，通过再分离获得的菌株发病症状也与初始病原菌症状一致，由此确定菌株21BS02-1即为致病菌。菌株21BS02-1的菌落主要呈白色，毡状，菌丝致密，菌落中央呈玫瑰色；其菌丝体细长，部分菌丝为有隔菌丝；大型分生孢子两端稍弯曲，形状似镰刀，其大小为(56.26~175.76)μm × (12.93~19.78)μm，有3~7个分隔；小型分生孢子较少，呈椭圆形，0~1个分隔；根据形态特征初步鉴定该病原菌为镰刀菌（Fusarium sp.）。进一步对菌株21BS02-1的ITS、β-tubulin、TEF基因序列进行克隆测序，经Blast搜索表明，其ITS、β-tubulin、TEF序列与Fusarium lateritium的同源性分别达到99.56%（MN686293）、100%（KJ00154）、99.68%（KF918550）。3个单基因聚类树均一致，表明菌株21BS02-1属于砖红镰刀菌（F. lateritium）。生物学特性测定表明，致病菌21BS02-1最适合的培养基为OMA和CMA培养基；碳源为蔗糖时的利用率最高；牛肉浸膏为致病菌最适生长的氮源；12 h光暗交替条件下菌丝生长最快；pH在7~9范围内最适合该致病菌生长。本研究是砖红镰刀菌（F. lateritium）侵染中粒种咖啡较为详细的报道。