禾长蠕孢菌和尖角突脐孢菌防治稗草的研究

 病原真菌禾长蠕孢菌稗草专化型(Helminthosporium gramineum Rabenh f.sp.echinochloae,HGE)和尖角突脐孢菌(Exserohilum monoceras,EM)在稗汁葡萄糖中的发酵滤液对稗草种子的发芽有明显的抑制效果,发芽抑制率分别为30.9%和13.5%.HGE菌在改良Fries、稗汁葡萄糖,EM菌在改良Fries中的发酵滤液对稗草根和芽的生长有明显抑制作用.HGE菌发酵滤液与其孢子混合使用比单独使用对稗草的防效明显提高,稗草感病株率、致死率分别达86.3%和69.5%,病情指数为78.7;发酵滤液与孢子结合(先后喷雾)使用后稗草的感病株率、致死率分别为83.9%和67.9%,病情指数为72.8.HGE与弯孢菌(Curvularia lunata)孢子混合喷雾接种对稗草防效明显高于2种菌孢子单独使用.     

影响链格孢菌生长及产孢的因子

在实验室研究了影响稗草病原真菌链格孢生长及产孢的因子.链格孢菌可在较宽的温度范围内生长、繁殖;连续黑暗、空气充足条件下有利于产孢;该菌在4种不同的植物产品上培养可产生大量的孢子,最多的无芒稗籽上每克干物质产孢量达3.59×108个,这些培养物在首次产孢收获后均可直接培养作二次产孢利用.固体(菌块)和液体(菌悬液)接种培养物均能成功产孢.但培养量大时液体接种优于固体接种,产孢量与接种菌悬液浓度呈正相关.     

应用均匀设计法筛选稗草病原菌产孢最佳配方培养基

 筛选生防菌最佳产孢培养基是高效、大量生产生防菌孢子的基础。应用均匀设计法结合二次多项式回归分析,从众多因素中筛选出稗草病原菌弯孢菌(Curvularia lunata)和互格交链孢菌(Alternaria alternata)最佳产孢培养基组分,建立多元二次回归数学模型。弯孢菌最佳产孢组分为胡萝卜、芹菜、欧芹、水芹、莴苣、蔗糖和Vc;互格交链孢菌最佳产孢组分为胡萝卜、芹菜、水芹、莴苣和蔗糖。根据回归模型计算理论(拟和)产孢值。比较实测产孢量和理论产孢值,结果表明,稗草病原菌的实测产孢量和回归模型拟合产孢值非常接近,拟合误差小。说明采用均匀设计法从众多因素中筛选培养基最佳成分是可行的。     

尖角突脐孢对稻田稗草的防除效果

在温室内进行了露水期、接种量、喷液量及稗草叶龄对尖角突脐孢菌(Exserohilummonoceras)除稗效果的试验.结果表明,适合条件下,尖角突脐孢对4叶期以下的稗草幼苗具有极好的防效.要达到理想的除稗效果需要至少24h的露水期,接种菌液的孢子浓度在1.25×105孢子/ml以上.接种孢子浓度和喷液量对尖角突脐孢除稗效果的影响也极明显,它们与防效呈S曲线关系.     

三株病原真菌对稗草生防潜力的研究

从稻田中采集自然感病的稗草标样,分离获得10余种病原真菌,经生测,链格孢菌、尖角突脐孢菌和内脐孢菌是3株很有希望的稗草生防潜力菌。对它们的生长及产孢特性、寄主范围及孢子悬浮液喷雾接种对水稻和稗草的致病性等进行了研究。结果表明,3个菌各有优缺点。链格孢菌对稗草致病力稍差,对水稻有轻度感染,但其产孢速度快,产孢量大;尖角突脐孢菌对稗致病力强,对稻无影响,但产生孢子慢,产生菌丝多;内脐孢菌对稗致病力强,对稻安全,菌落生长较慢,但产孢速度快,产孢量多,气生菌丝少。黑暗、空气充足有利于3菌株的生长及产孢。     

大批量生产百日草链格孢菌孢子的技术

通过研究光照、菌丝体粉碎、培养基等因素对百日草链格孢菌孢子形成的影响，试图探寻大批量生产该菌孢子的条件和方法。结果显示，低氮培养、近紫外光照射、适当适时的菌丝体粉碎能诱导百日草链格孢菌形成大量的孢子。在上述条件下，应用液体发酵培养菌丝体，结合固体培养诱导孢子，是一种可行的方法。     

稗草生防菌新月弯孢菌Culvularia lunata菌株J15(2)的安全性和致病力

为了确定生防菌新月弯孢菌Culvularia lunata菌株J15(2)的安全性及防除稗草的潜力,研究了该菌株的寄主范围,接种孢子时稗草叶龄、接种后保湿时间、接种浓度对该菌株致病力的影响,土壤带菌对稗草出苗的影响,以及与化学除草剂的相互作用.结果显示,菌株J15(2)只能在稗草上侵染并扩展;接种孢子后保湿24 h,对2叶期稗草抑制率可达55.6%,对1.5叶期稗草抑制率可达100%;保&48h,对2叶期稗草防效提高至91.6%;接种孢子量需达1012孢子/hm2才能较好地抑制稗草生长;土壤带菌亦可抑制稗草生长.该菌与化学除草剂精噁唑禾草灵或二氯喹啉酸混用能显著提高除稗效果,菌株J15(2) 3.3 × 1012孢子//hm2分别与精噁唑禾草灵有效剂量3.83g/hm2二氯喹啉酸有效剂量250.00g/hm2混用,对稗草防效分别为99.5%和88.6%,远高于这两个除草剂单用时的药效.该试验结果表明,菌株J15(2)具有作为微生物除草剂的开发潜力.     

尖角突脐孢菌在温室环境下侵染稗草的致病体系研究

稗属（Echinochloa）杂草是全世界水稻等作物生产中的一类恶性杂草。人工除稗安全但费时费力，化学除稗高效但伴随农产品安全和环境相融友好方面的潜在风险。因此，寻求安全高效的除稗活性成分和施药技术成为国际上除稗剂发展的重要方向之一。在研究和开发生物除稗剂的技术体系中，满足高通量筛选安全、高效的除稗生防因子的温室检测体系是极为重要的支持平台，同样，生物除稗剂的田间科学施药技术亦应依据生防因子在稗草上致病和致害所需的最佳条件来制定。基于此，亟需探明生防因子侵染稗草致病的相关条件并建立快速有效的生防因子除稗效果温室检测体系和田间施药技术。本文系统研究了影响尖角突脐孢菌侵染稗草的相关因子，建立了稳定的温室环境下尖角突脐孢菌侵染稗草的致病体系。     

一种筛选稗草生防潜力菌的简易生测方法

将催芽至露白的稗草和水稻种子直接接种在于PDA培养基上培养形成的 4种稗草病原菌 (交链孢菌、弯孢菌、禾长蠕孢菌稗草专化型、尖角突脐孢菌 )菌落上 ,观察稻、稗苗感病情况 ,调查根、芽生长抑制率。结果表明 ,4种菌对稻、稗的致病性和生长抑制率与常规生测法结果基本吻合。说明在培养基中初步测定、筛选稗草生防潜力菌是可行的。该方法与常规生测法比较 ,可节省一半时间左右 ,同时可减少工作量。     

木耳菌糠培养球孢白僵菌的条件筛选及优化

白僵菌是一种虫生真菌,目前在农林害虫防治中取得了明显的效果。为了进一步促进白僵菌农药的发展,开发低成本的白僵菌发酵方法或优化其发酵条件是不可缺少的一环。本文尝试以木耳菌糠作为主要培养基液体发酵球孢白僵菌,通过单因素试验和正交试验确定菌糠培养基配方并优化发酵条件。结果表明,废弃菌糠适宜作为球孢白僵菌的主要培养基,当料液比为1:4,补充0.3%硫酸锰、0.5%硝酸铵,接菌量10%,培养温度为25℃,培养时间为4 d时,球孢白僵菌的产孢量达到了2.0×109个孢子/mL。研究证实了木耳菌糠培养球孢白僵菌的可行性,希望为低成本发酵其他生防菌提供新思路。     

具有除草活性的小麦根腐病菌发酵条件优化

通过单因子试验和正交试验对小麦根腐病菌菌株进行了发酵条件优化,以提高其活性物质的产量。结果表明:最适合小麦根腐病菌产毒的培养基是察氏培养基;该培养基的最佳碳源为葡萄糖,氮源为氯化铵,无机盐为氯化钾、磷酸氢二钾、硫酸镁和硫酸亚铁;发酵培养基各组分的最佳配比为:葡萄糖2.0%,氯化铵0.25%,氯化钾0.05%,磷酸氢二钾0.025%,硫酸镁0.025%,硫酸亚铁0.000 5%;有利于提高菌株活性物质产量的培养条件为初始pH7.0,转速为130r/min,温度为28℃,培养时间为5d,装液量为120mL/250mL,接种量为6%。小麦根腐病菌菌株发酵产物具有良好的热稳定性和光稳定性,在弱酸性及中性条件下较稳定。发酵条件优化后提高了小麦根腐病菌的除草活性。     

粉红螺旋聚孢霉NF-06固体发酵条件优化及对南方根结线虫的防治效果

南方根结线虫是一种严重危害我国蔬菜安全生产的土传病害，生产上亟需一种有效的防控方法。本研究从根结线虫卵囊中分离到一株生防真菌NF-06，通过形态学和分子生物学方法将其鉴定为粉红螺旋聚孢霉Clonostachys rosea。通过离体活性测定的方法测定NF-06发酵滤液对根结线虫2龄幼虫的致死作用，发现24、48和72 h后2龄幼虫的相对死亡率分别达63.6%，75.5%和87.0%。以产孢量为指标，优化菌株NF-06固体发酵条件，确定最佳基础培养基为麦麸和麦秸秆混合物，其比例为4：1（wt/wt），在基础培养基中添加10%玉米粉、2%蚕豆粉和0.05% KH₂PO₄可获得最大产孢量；最优培养条件为初始pH值7.0，初始含水量40%，接种量5%，产孢量达8.9×10¹⁰ CFU/g。室内盆栽防治效果测定显示，菌株NF-06固体发酵物能够有效降低土壤中的线虫数量和番茄根结指数，其中5 g/株穴施处理的线虫减退率和防治效果分别达67.9%和69.6%。本研究为生物杀线虫剂的开发和利用提供理论依据。     

不同培养基对玫烟色棒束孢菌株的复壮研究

虫生真菌的毒力退化是一种普遍现象,表现为产孢能力下降或丧失、毒力下降及次级代谢产物合成能力下降等.基于玫烟色棒束孢Isaria fumosorosea PF904菌株继代培养后产孢量和毒力严重退化,本试验采用添加4种不同营养物质的培养基对其进行复壮.结果 表明:培养基中添加甲壳素粉后菌株毒力恢复最好,复壮第10代时对小...     

Selection and orchard testing of antagonists suppressing conidial production by the apple scab pathogen <Emphasis Type="Italic">Venturia inaequalis</Emphasis>

Apple scab caused by Venturia inaequalis is a major disease in apple production. Epidemics in spring are initiated by ascospores produced on overwintering leaves whereas epidemics during summer are driven by conidia produced on apple leaves by biotrophic mycelium. Fungal colonisers of sporulating colonies of V. inaequalis were isolated and their potential to reduce the production of conidia of V. inaequalis was evaluated on apple seedlings under controlled conditions. The four most effective isolates of the 63 screened isolates were tested subsequently under Dutch orchard conditions in 2006. Repeated applications of conidial suspensions of Cladosporium cladosporioides H39 resulted in an average reduction of conidial production by V. inaequalis of approximately 40%. In 2007, applications of conidial suspensions of C. cladosporioides H39 reduced conidial production by V. inaequalis by 69% on August 6 and by 51% on August 16, but no effect was found on August 20. However, viability of available conidia of C. cladosporioides H39 was low at the end of the experiment. Epiphytic and endophytic colonisation by Cladosporium spp. of leaves treated during the experiment with C. cladosporioides H39 was significantly higher than on control leaves sampled 6 weeks after the last application. It is concluded that C. cladosporioides H39 has promising potential as a biological control agent for apple scab control. More information is needed on the effect of C. cladosporioides H39 on apple scab epidemics as well as on mass production, formulation and shelf life of conidia of the antagonist.     

两株哈茨木霉菌株防治水稻纹枯病及促进水稻生长的潜力研究

 由茄丝核菌(Rhizoctonia solani)引起的纹枯病是水稻生产中的重要真菌病害,严重影响水稻产量及品质。本研究评估了两株哈茨木霉菌株3S1-13和4S2-46对纹枯病生防潜力及其对水稻种子萌发和幼苗生长的促生效果。研究表明两株哈茨木霉菌株在马铃薯葡萄糖琼脂培养基(PDA)上不产生分生孢子,但在含有1%酵母粉的PDA上能恢复产分生孢子的能力。菌株3S1-13和4S2-46发酵液对茄丝核菌的菌丝生长及菌核形成均有较强抑制作用,具有防治水稻纹枯病潜力,且发酵液防效显著高于孢子液的防效。菌株3S1-13和4S2-46发酵液处理后的水稻种子发芽率和幼苗的根系、鲜重均显著高于对照,显示出有利于促进种子发芽与生长的作用。研究结果表明两株哈茨木霉菌株发酵液不仅可以用于水稻纹枯病的生物防治,而且可以促进水稻种子发芽及生长,具有较好的应用潜力。     

深绿木霉TA-9分生孢子固体发酵的响应曲面法优化

前期研究中分离获得深绿木霉Trichoderma atroviride菌株TA-9,其在田间试验中具有较好生物防治效果.本研究通过Plackett-Burman试验设计及Box-Behnken设计-响应曲面法(response surface methodology,RSM)对影响深绿木霉TA-9菌株发酵的培养基和培养...     

Production, Survival, and Evaluation of Solid-Substrate Inocula of Penicillium oxalicum, a Biocontrol Agent Against Fusarium Wilt of Tomato

ABSTRACT Production of conidia of Penicillium oxalicum (ATCC number pending), a biocontrol agent of Fusarium oxysporum f. sp. lycopersici, was tested in liquid and solid fermentation. P. oxalicum produced 250-fold more conidia in solid than in liquid fermentation at 30 days after inoculation of substrate. Solid fermentation was carried out in plastic bags (600 cm(3)) especially designed for solid fermentation (VALMIC) containing 50 g of peat/vermiculite (PV) (1:1, wt/wt) with 40% moisture, sealed, sterilized, and then inoculated with 1 ml of a conidial suspension of P. oxalicum (10(5) conidia g(-1) dry substrate), sealed again, and incubated in darkness at 20 to 25 degrees C for 30 days. Addition of amendments to PV in a proportion of 0.5 (wt/wt) significantly increased conidial production of P. oxalicum. The best production was obtained on PV plus meal of cereal grains (barley) or leguminous seeds (lentil) (100-fold higher). Conidial production obtained after 5 days of inoculation was similar to that obtained at 30 days. However, viability of conidia produced in PV plus lentil meal was 35% higher than that of conidia produced in PV plus barley meal. Changes in proportions (1:1:0.5, wt/wt/wt; 1:1:1, wt/wt/wt; 1:0.5:0.5, wt/wt/wt; 1:1:0.5, vol/vol/vol) of components of the substrate (peat/vermiculite/lentil meal) did not enhance production or viability of conidia. Optimal initial moisture in the substrate was 30 to 40%. At lower moistures, significant reductions of production of conidia were observed, particularly at 10%. There was a general decline in the number of conidia in bags with time of storage at -80, -20, 4, and 25 degrees C, or at room temperature (range from 30 to 15 degrees C), with the highest decline occurring from 60 to 180 days. Conidial viability also was reduced with time, except for conidia stored at -20 degrees C. Fresh conidia produced in solid fermentation system or those conidia stored at -20 degrees C for 180 days reduced Fusarium wilt of tomato by 49 and 61%, respectively.     

Solid substrate for production of Alternaria alternata conidia: a potential mycoherbicide for the control of Eichhornia crassipes (water hyacinth)

Eichhornia crassipes (water hyacinth), a major aquatic weed in India, is one of the targets in a biological weed control research programme in India. A fungal pathogen, Alternaria alternata, is being evaluated as a biological control agent for this weed. The feasibility of solid substrate for the mass production of A. alternata has been examined. Conidia production and virulence of A. alternata were affected by temperature, light and incubation period. The highest number of conidia were produced on rice seed followed by wheat, sorghum, maize seeds and cornmeal at 20°C when exposed to near‐ultraviolet than on the other substrates, while the least conidia were observed on these substrates under light conditions. At 20°C, large numbers of virulent conidia were produced on rice seeds after 4 weeks of incubation under constant dark conditions. Henceforth, the use of rice seeds as a solid substrate for production of A. alternata could be a feasible method to produce conidia in a village co‐operative scenario in India.     

Use of sclerotia of<Emphasis Type="Italic">Sclerotinia sclerotiorum</Emphasis> for efficient production of conidia of<Emphasis Type="Italic">Coniothyrium minitans</Emphasis> in liquid culture

A study was conducted to determine the feasibility of using sclerotia ofSclerotinia sclerotiorum for producing conidia ofConiothyrium minitans in liquid culture. The medium (SST) was made of water containing 2.0, 1.5, 1.0 or 0.5% (w/v) ground sclerotia ofS. sclerotiorum and 100 μgl ⁻¹ thiamine hydrochloride (HCl). One ml of conidial suspension (2 × 10⁷ conidia ml⁻¹) ofC. minitans LRC 2534 was inoculated into 100 ml of SST medium or control (thiamine HCl in water) and incubated at 20 ± 2°C on a shaker at 200 rpm. Subsamples were removed periodically and examined under a compound microscope. Conidia in the SST media germinated within 24 h, developed into branched hyphae within 48 h, produced pycnidia after 3–4 days, and the pycnidia released mature conidia after 7 days. Production of conidia varied with the concentration of sclerotia in the SST medium. It was lower (3.6 × 10⁶ conidia ml⁻¹) at 0.5% but higher (1.2 × 10⁸ conidia ml⁻¹) at 2%. The new conidia were viable and the colonies developing from them showed the original morphological characteristics. It was concluded that using SST liquid medium as a substrate for mass production of conidia ofC. minitans has potential for use in commercial development of this mycoparasite as a biocontrol product. http:www.phytoparasitica.org posting Jan. 23, 2007.     

The effect of temperature and water potential on the production of conidia by sclerotia of Botrytis squamosa

The rate of conidiogenic germination of Botrytis squamosa was highest at 16°C and the greatest numbers of conidia per sclerotium (up to 5 × 10⁴) were produced at temperatures of 5–10°C. At temperatures above 20°C, the percentage of sclerotia producing conidia declined rapidly. Decreasing water potential reduced the rate at which conidia were produced and also resulted in fewer conidia produced per sclerotium. However, conidia were produced at water potentials as low as −2 MPa, at which sclerotial germination was at least 60%. A simulation model that included effects of both temperature and water potential was developed from laboratory and field data obtained for conidial production in sclerotia exposed for periods of 1, 2, 3 or 4 weeks during an entire year. There was good agreement between conidiogenic germination predicted by the model and conidial production observed in onion plots artificially inoculated with sclerotia. Temperature and water potential were therefore considered to be the principal microclimatic factors affecting conidial production by B. squamosa. The role of sclerotia in the context of UK onion production is discussed.