Management of Strawberry Grey Mold Disease Using Biocontrol Agents and Plant Extracts

Strawberry (Fragaria × ananassa Duch.) is a significant global soft fruit crop, prized for its nutrient content and pleasant flavor. However, diseases, particularly grey mold caused by Botrytis cinerea Pers. Fr. poses major constraints to strawberry production and productivity. Grey mold severely impacts fruit quality and quantity, diminishing market value. This study evaluated five B. cinerea isolates from various locations in the Ri-Bhoi district of Meghalaya. All isolates were pathogenic, with isolate SGM 2 identified as highly virulent. Host range studies showed the pathogen-producing symptoms in the fava bean pods, marigold, gerbera, and chrysanthemum flowers and in the fava bean, gerbera, and lettuce leaves. In vitro tests revealed that neem extract (15% w/v) achieved the highest mycelial growth inhibition at 76.66%, while black turmeric extract (5% w/v) had the lowest inhibition at 9.62%. Dual culture methods with bio-control agents indicated that Bacillus subtilis recorded the highest mean inhibition at 77.03%, while Pseudomonas fluorescens had the lowest at 20.36% against the two virulent isolates. Pot evaluations demonstrated that B. subtilis resulted in the lowest percent disease index at 20.59%, followed by neem extract at 23.31%, with the highest disease index in the control group at 42.51%. Additionally, B. subtilis significantly improved plant growth, yielding an average of 0.32 kg compared to 0.14 kg in the control. The promising results of B. subtilis and neem leaf extract from this study suggest their potential for eco-friendly managing grey mold in strawberries under field conditions.

Trichoderma viride Strains against Vegetable Grey Mold in Greenhouse

[Objective] The paper aimed to study the control effects of live spore preparations of Trichoderma viride strains against vegetable grey mold in greenhouse. [Method] Trichoderma viride strains NW-411 live spore preparations were prepared by solid-state fermentation,106-107 spore/g diluent was made to conduct field control experiment,traits change of cucumber and tomato plants inoculated grey mold were investigated,control effect was calculated. [Result] Cucumber and tomato plants without dilution treatment of T. viride spores could be infected with different changes in trait. T. viride spore preparations had a significant preventive effect on greenhouse cucumber and tomato gray mold,the optimal concentration of spores was in the range of 2.3×10^6-2.3×10^7 spore/g. The incidence of cucumber and tomato plants were reduced to 4.2% and 3.1%,the incidence rate decreased 29.8% and 39.1% compared with plants without treatment,biological control effect was over 87%,and the plant growth can be enhanced obviously. [Conclusion] Live spores preparation of T. viride not only had a significant effect on grey mold,but also significantly enhanced the plants growth in greenhouse,which is a safety and environmental protection biological agent,and worthy to be widely spread in large-scale green vegetable production.

盾叶薯蓣灰霉病(Botrytis cinerea)的研究

对一种引起盾叶薯蓣叶、茎呈暗绿色至黑褐色水渍状软腐的病害进行了调查 ,并作了鉴定。结果明确其为灰葡萄孢菌 (BotrytiscinereaPers.exFr )引起的一种新病害 ,根据症状和病原鉴定将该病称为盾叶薯蓣灰霉病。该病原菌生长温度 0～ 30℃ ,最适生长温度 10～ 2 5℃。病害在每年春季、夏初和秋末 ,雨水多的年份发生严重。

连续变温对番茄灰霉病(Botrytis cinerea)致病性的影响

为探究连续变温对番茄灰霉病致病性影响,以中抗性品种Money Maker为供试植株,在离体叶片上接种,在RH55%,RH75%,RH85%和RH95%条件下分别设置12个变温处理,研究接种叶片在5d内病斑生长状况。接种后24h开始观察,每隔12h观测1次,以病斑面积和平均生长速度作为衡量依据。结果表明:培养5d后RH55%、RH75%和RH85%条件下10h和12h处理病斑面积明显大于同湿度其他处理;1,2,4,18,20和22h处理生长量出现不同程度的抑制,最大抑制作用可达到70%以上。在RH95%条件下1,2,4h处理生长情况明显低于同湿度其他变温处理,但18,20,22h没有抑制作用。这种抑制作用与低温或高温时间在一天当中所占的比例有关,24h内温度低于15℃的时间长于18h或温度高于25℃时间长于6h都会对菌丝产生抑制作用。

人参灰霉病菌Botrytis cinerea的种群表型和基因型多样性

为探究人参灰霉病菌Botrytis cinerea的种群表型和基因型多样性,对从我国人参主产区的8个地区所分离到的对人参有致病性的154株灰葡萄孢菌株进行研究,在PDA上培养3周观察其表型,通过PCR扩增的方法鉴定其转座因子类型和MAT基因型,并对不同转座因子类型的菌株进行致病力的测定。通过试验得到8种表型和3种转座因子型(Boty、Flipper和Transposa),不存在Vacuma类型,其中Transposa类型所占比例最高为93.51%,且致病力最强;同时也发现了含MAT1-1和MAT1-2的菌株个数分别为73株和75株,2种基因型在该种群中均匀分布。以上研究结果表明,我国人参灰霉病菌B.cinerea具有相对均匀的种群多样性,同时也具有丰富的表型和遗传分化。

Isolation and Identification of Botrytis cinerea in Processing Tomato(Solanum lycopersicum)

[Objective]The paper was to isolate and identify Botrytis cinerea from processing tomato.[Method]The strain nky007a was isolated and purified from fruits infected by tomato grey mold.After identification,it was determined as B.cinerea.The 18S rDNA and internal transcriptional spacer(ITS)sequences of strain nky007a were amplified for molecular identification and classification.[Result]The 18S rDNA sequence of the strain shared the homology of 99.95%with Botryotinia fuckeliana(EF 110887.1);the ITS rDNA sequence had the homology of 100%with Botryotinia fuckeliana(AB444949.1),and there was no difference in bases.Phylogenetic tree analysis of strain nky007a indicated that nky007a belonged to Botrytis.When healthy tomatoes were infected by the strain,the incidence rate of tomato grey mold was up to 95%,indicating the pathogenicity of the strain.[Conclusion]Morphological identification,molecular identification and pathogenicity analysis results show that the strain nky007a is a highly pathogenetic strain B.cinerea.

Production and Testing of Biopesticide for Control of Postharvest Mold Infections on Fresh Fruits of Apple and Pear

The present research aimed at producing a biopesticide with proper formulation of invert emulsion (water-in-oil type) and effective strain of?Trichoderma harzianum?then, testing it against pathogens of postharvest mold infections(Botrytis cinerea?&?Penicillium expansum) on fresh fruits of apple and pear. The?proper formulation of invert emulsion (IE#3) used for biopesticide production has the following ingredients (100% w/w): soybean oil (28.50%), coconut oil (19.50%), oil-soluble emulsifier: Tween 20 (2.0%), glycerine (4.25%), water-soluble emulsifier:?dehymuls k (0.75%),?sterile distilled water (22.5%) and conidial suspension of the effective strain ofT. harzianum?“TrichoPAL1” in water (22.5%, concentration 1 ×?107?conidia/ml). Testing efficacy of the produced biopesticide has indicated a significant reduction in the disease lesion diameter of mold infections on wounded apple and pear fruits stored at?20℃?± 1℃?compared to the untreated fruits or control (reduction from up to 38.75 to about 7.50 mm, respectively, according to?the type of mold infections and fruit type). Also, the treatment with the produced biopesticide has resulted in?a long protection period from mold infections on wounded and un-wounded fresh fruits of apple and pear stored under controlled and semi-commercial conditions(up to 2.5 months according to the type of mold infections and fruit type). In conclusion, the overall results have demonstrated the effectiveness of produced biopesticide on stored fruits under controlled and semi-commercial conditions therefore,?it is recommended to test this effectiveness on marketed fruits stored under variable conditions before applying it at a large scale.

Field Control Effects of Four Biopesticides on Grape Gray Mold

[Objectives]The paper was to search for biopesticides with good control effect on grape gray mold.[Methods]A randomized field trial was carried out to compare the control effects of 100 million CFU/g Trichoderma harzianum WDG,200 million spores/g Trichoderma WP,24% validamycin AS,0.3% matrine AS on grape gray mold.[Results]Of the 4 biopesticides,200 million spores/g Trichoderma WP had the highest control effect on grape gray mold,with the control effect of 73.38% at 7 d post the second application,significantly higher than those of 100 million CFU/g T.harzianum WG and 0.3% matrine AS;the control effect was 75.77% at 14 d post the second application,significantly higher than those of other 3 biopesticides.[Conclusions]The 200 million spores/g Trichoderma WP shows a good control effect on grape gray mold with a long-lasting control effect,so it is more is for the control of summer black grape gray mold in Jianshui County.

河南省番茄灰霉病病原菌鉴定、生物学特性分析及杀菌剂筛选

番茄灰霉病是番茄生产上的重要病害之一。本试验以河南商丘市蔬菜大棚内疑似患灰霉病番茄植株叶片为试材,采用组织分离法纯化获得3株病原菌菌株,其形态特征一致,选择一株代表性菌株命名为SQBC1。对该菌株rDNA-ITS序列、CHS-1、tubA基因进行测序、比对和系统进化分析,结合菌株培养性状、显微形态学观察、致病性分析,将河南商丘番茄灰霉病致病病原菌确定为灰葡萄孢菌Botryotinia cinerea。生物学特性研究表明28℃为最适生长温度,12 h光照12 h黑暗为最适光照条件,6.0为最适pH值,乳糖为最适碳源,蛋白胨为最适氮源。测试了7种杀菌剂对灰霉菌菌丝生长的抑菌效果,其中嘧菌酯和吡唑醚菌酯的抑菌效果最好。该研究为番茄灰霉病的有效防治提供一定的参考依据。

几种杀菌剂对番茄灰霉病菌的抑菌效果对比研究

采用番茄灰霉病防治中6种已登记杀菌剂和7种未登记杀菌剂对番茄灰霉病菌进行室内毒力测定,并通过EC50和病原菌的抗性水平进行抑菌效果对比。结果表明:在已登记的6种杀菌剂中,以25%啶菌恶唑EC的抑菌效果最好,EC50值为0.14 mg.L-1番茄灰霉病菌对其表现敏感;在未登记的5种杀菌剂(包括10%多抗霉素B)中,25%咪鲜胺EC的抑菌效果最好,EC50值为0.66 mg.L-1番茄灰霉病菌对其表现敏感;在2种未登记活体微生物杀菌剂中,抑菌效果最好的为2×108cfu.g-1绿色木霉菌WP,EC50值为0.001×108cfu.L-1。

不同生物杀菌剂对人参灰霉病的室内毒力及田间防效

采用菌丝生长速率法测定了14种生物杀菌剂对人参灰霉病菌的室内毒力,并进行了7种生物杀菌剂对人参灰霉病的田间药效试验。室内毒力测定结果表明：0.3%丁子香酚 SL 和2%蛇床子素 EC 对人参灰霉病菌毒力最强,其 EC50分别为0.129、0.623 mg/L,2%春雷霉素 AS 等5种生物杀菌剂对人参灰霉病菌也具有较好的抑制效果,其 EC50在12.857～127.012 mg/L 之间。田间试验结果表明：0.3%丁子香酚 SL 在田间对人参灰霉病表现较高的防效,2%春雷霉素 AS 和3%中生菌素 WP 次之。初步筛选出了对人参灰霉病具有较好防效的生物杀菌剂,为人参灰霉病的安全防控提供科学依据。

甲基营养型芽孢杆菌BH21对葡萄灰霉病菌的拮抗作用

【目的】甲基营养型芽孢杆菌(Bacillus methylotrophicus)BH21是一株对葡萄灰霉病菌(Botrytis cinerea)有较好拮抗作用的海洋源细菌,鉴定该菌株脂肽类抗菌物质合成基因,检测脂肽粗提物对葡萄灰霉病菌的拮抗作用,为应用该菌株防治葡萄灰霉病提供科学依据。【方法】通过特异引物的基因组PCR法检测甲基营养型芽孢杆菌BH21菌株合成脂肽的能力;盐酸沉淀和甲醇抽提法从无菌发酵液中提取脂肽粗提物;排油圈法检测脂肽粗提物的表面活性;采用菌丝生长速率法检测脂肽粗提物对灰霉病菌菌丝生长的抑制作用并计算有效中浓度EC50;利用高效液相色谱技术(HPLC)对脂肽粗提物洗脱分离,并采用菌丝生长速率法检测各组分对葡萄灰霉病菌的抑制能力;采用反相高效液相色谱(RP-HPLC)分析主要抑菌成分的脂肽类型。采用离体叶片接种法检测脂肽粗提物对葡萄灰霉病的防治效果。【结果】选取11对特异引物对菌株BH21基因组扩增,其中7对引物扩增出预期核酸片段;扩增产物经测序、BLAST比对分析,结果显示扩增产物与相关菌株脂肽基因相似度为96%—99%,扩增产物翻译的蛋白与相关菌株的脂肽合成蛋白相似度为96%—100%,表明甲基营养型芽孢杆菌BH21基因组中含有ituA、bamD、ituC、ituD、fenD、srfAB、yndJ,该菌株具有合成surfactin、iturin及fengycin等多种脂肽类抗菌物质的能力。盐酸沉淀和甲醇抽提从LB无菌发酵液中获得脂肽粗提物,得率为428 mg·L^(-1)。排油圈检测结果显示,脂肽粗提物使橄榄油膜形成排油圈,表明脂肽粗提物具有表面活性。脂肽粗提物对葡萄灰霉病菌菌丝生长具有显著的抑制作用,脂肽粗提物浓度为440μg·mL^(-1)时,对葡萄灰霉病菌菌丝生长的相对抑制率为82.8%。根据毒力方程计算,抑制葡萄灰霉病菌菌丝生长的脂肽粗提物EC50为144.39μg·mL^(-1)。HPLC分离纯化脂肽粗提物获得6个组分,只有组分BH21-2和BH21-3抑制葡萄灰霉病菌的生长,RP-HPLC色谱图分析表明组分BH21-2和BH21-3属于fengycin家族脂肽。葡萄灰霉病离体叶片试验结果表明,脂肽粗提物浓度为400μg·mL^(-1)时,对葡萄叶片灰霉病防病效果为100%;脂肽粗提物浓度为220μg·mL^(-1)时,对葡萄叶片病斑扩展相对抑制率为94.4%。【结论】甲基营养型芽孢杆菌菌株BH21具有合成surfactin、iturin及fengycin等多种脂肽类抗菌物质的基因,该菌株脂肽粗提物对葡萄灰霉病菌具有较强的拮抗作用,在葡萄灰霉病生物防治中具有应用潜力。

番茄灰霉病防治研究进展

由灰葡萄孢引起的灰霉病是一种世界性病害,该病菌可引起多种作物的灰霉病,尤其在番茄上危害较为严重。本研究从番茄灰霉病的发生及为害、防治对策、国内外对番茄灰霉病的研究现状、防治番茄灰霉病的趋势及展望4个方面,对当前番茄灰霉病的防治研究进展进行了综述。

葡萄灰霉病无公害防治研究进展

葡萄灰霉病是葡萄生产中最常见、危害最严重的病害之一,主要由灰葡萄孢菌(Botrytis cinerea Pers.)引起。概述了葡萄灰霉病病原菌发病规律及侵染特征,重点阐述了葡萄灰霉病无公害防治的研究进展,并对当前研究与应用中存在的问题进行了探讨,以期为该病害的深入研究和有效防治提供参考。

基于可见/近红外光谱技术的茄子叶片灰霉病早期检测研究

应用可见/近红外光谱技术对茄子叶片进行灰霉病害还未在叶片表面出现病症时的早期检测.采用化学计量学方法建立早期检测模型.主成分分析用于对光谱数据进行降维,得到若干个最重要的主成分.但直接从聚类图中无法进行是否染病的鉴别.因而将其作为变量输入BP神经网络,从而减少了计算量,提高了建模精度.检测结果显示,模型具有良好的检测效果,能够达到100%的识别率,正确率也能达到88%.说明运用可见/近红外光谱技术能够实现当病症还未在叶片表面出现时的快速准确的早期检测,为灰霉病早期检测提供了新的途径.

创制杀菌剂啶菌恶唑的生物活性及应用研究(Ⅱ)黄瓜灰霉病

啶菌恶唑是一种结构新颖、作用方式独特的新型杀菌剂,具有保护、治疗效果及良好的内吸传导性。对子囊菌、担子菌和半知菌引起的多种植物病害具有良好的防治作用。啶菌恶唑对黄瓜灰霉病具有很高的杀菌活性,室内离体试验结果啶菌恶唑对黄瓜灰霉病 (Botrytis cinerea Pers) 的EC50值为0.199μg/ml;1998～2002年多点的田间小区试验结果表明:啶菌恶唑对黄瓜灰霉病具有良好的防治效果,在200～400 g a.i./hm2处理剂量下、施药2～3次、间隔期6～8d可有效控制黄瓜灰霉病的发生与危害。防效优于目前生产上推广使用的多菌灵、腐霉利、菌核净、嘧霉胺等常规杀菌剂。

猕猴桃灰霉病病原菌鉴定及室内药剂筛选

通过病原菌形态特征、致病性测定及rDNA-ITS的同源性分析,确定江西奉新猕猴桃灰霉病的病原菌为Botry-tis cinerea（有性态Botryotinia fuckeliana）,该病菌在5～30℃的温度范围内均能正常生长,且该病菌只能通过伤口接种引起猕猴桃发病.采用菌丝生长速率法测定11种杀菌剂对该病原菌的毒力大小,结果表明,肟菌·戊唑醇水分散粒剂、氟硅唑乳油、异菌脲悬浮剂、苯醚甲环唑水分散粒剂、戊唑醇悬浮剂和吡唑醚菌酯乳油对其菌丝生长有较强的抑制作用,ECs0值分别为0.118 0、0.288 6、0.355 2、1.541 8、1.760 2和1.778 4 μg/mL.

黑龙江省番茄灰霉病病原菌的分离与鉴定

为了对病原菌进行形态学观察及ITS序列鉴定,以黑龙江省齐齐哈尔市某温室中疑似感染番茄灰霉病的番茄植株叶片为试材,采用组织分离方法,分离并纯化病原菌。结果表明,共分离到4株病原菌,根据形态学特征及ITS序列系统进化分析结果,明确了引起番茄灰霉病的病原菌为灰葡萄孢菌(Botrytis cinerea)。

草莓灰霉病菌对速克灵的抗性研究

速克灵对草莓灰霉病具有很好的防治效果,但长期、单一使用很容易使草莓灰霉病菌产生抗药性。为合理使用这类杀菌剂,延缓或防止抗药性的产生,笔者通过UV-C诱变草莓灰霉病菌敏感菌株CF1获得了对速克灵具有高抗性的突变株CF1-1和CF1-2,并对抗性突变株的生长量、产孢量和致病力进行了研究,结果表明:CF1-1与CF1-2对速克灵抗性程度高,抗性水平与敏感菌株相比分别增加了484.6倍和911.0倍。抗性菌株在无速克灵的培养基上菌丝生长较敏感菌株弱,产孢量也明显少于敏感菌株;但其引起草莓离体叶片发病较敏感菌株重,病斑大且边缘颜色深;薄片黄瓜测定结果也表明,在无速克灵的情况下,敏感菌株CF1引起发病较重;而在有药条件下CF1受到一定抑制,但抗性菌株仍能致黄瓜片发病。将抗性菌株连续培养7代后对速克灵的抗性仍是相对稳定的。

枯草芽孢杆菌可湿性粉剂防治黄瓜灰霉病药效试验

采用室内生物活性测定及田间药效试验,以40%嘧霉胺悬浮剂为对照药剂,评价了生物杀菌剂枯草芽孢杆菌可湿性粉剂(活芽孢数为1 000亿.g-1)对黄瓜灰霉病的防治效果。试验结果显示,枯草芽孢杆菌可湿性粉剂对黄瓜灰霉病的抑制效果较好,与对照药剂相当,其EC50为5.89×106cfu.mL-1。田间试验中枯草芽孢杆菌可湿性粉剂0.08 g.m-2施药后的防效为70.81%,对照药剂0.11 g.m-2的防效为77.40%。室内生物活性测定结果与田间试验结果一致。