Use of carbendazim and carbendazim-resistant yeasts to create different yeast densities on wheat leaves for field studies on biological control

Phyllosphere yeasts may play an important role in suppressing necrotrophic pathogens. In a randomized block design, yeast densities on flag leaves and second leaves of field-grown wheat were left unchanged (D), or were lowered by sprayings with carbendazim (A), or were raised by sprayings with a mixture ofSporobolomyces roseus, Cryptococcus laurentii var.flavescens and nutrients (B), or with carbendazim, carbendazim-resistantS. roseus andC. laurentii var.flavescens and nutrients (C). The application of carbendazim to plots A and C would avoid a biased interference of carbendazim due to other effects than reducing the natural yeast population. Prolonged differences in yeast density would be reflected in differences in severity of diseases caused by carbendazim-insensitive necrotrophic pathogens.Both treatments B and C enhanced the yeast density about 10-fold at the beginning of leaf colonization, 2 to 3 weeks after leaf emergence. Untreated leaves (D), however, were rapidly leaves. Initially, the yeasts were suppressed by carbendazim (A). However, progressively the yeast population consisted of carbendazim-resistant strains which made the carbendazim sprayings ineffective. Therefore, the substantial differences in yeast density were of limited duration. The effect of the treatments on naturally occurringMycosphaerella graminicola (anam.Septoria tritici), Puccinia recondita f. sp.tritici and the total necrotic leaf area (D-value) is discussed.Samenvatting Fyllosfeergisten kunnen misschien een belangrijke rol spelen in de onderdrukking van necrotrofe pathogenen. In een blokkenproef werd de dichtheid van gisten op vlagbladeren en tweede bladeren niet beïvvloed (D) of verlaagd door bespuitingen met carbendazim (A), of verhoogd door bespuitingen met een mengsel van gisten (Sporobolomyces roseus enCryptococcus laurentii var.flavescens en voedingsstoffen (B), of met een mengsel van carbendazim, carbendazim-resistenteS. roseus enC. laurentii var.flavescens en voedingsstoffen (C). De grootste verschillen in dichtheden van de gistpopulaties werden verwacht tussen de behandelingen A en B of C. Door alle veldjes A en C met carbendazim te behandelen wordt een eenzijdig effect van carbendazim, berustend op andere eigenschappen dan de onderdrukking van de natuurlijke gistpopulatie, voorkomen. Langdurige verschillen in de populatiedichtheden van de gisten zouden weerspiegeld kunnen worden in verschillen in aantasting door carbendazimongevoelige necrotrofe pathogenen.Aan het begin van de bladkolonisatie door gisten, 2 tot 3 weken na het verschijnen van het blad, leidde zowel behandeling B als C tot, een 10-voudige verhoging van de gistdichtheid. De onbehandelde bladeren (D) werden echter spoedig door de van nature voorkomende gisten gekoloniseerd en het verschil tussen de onbehandelde en met gist bespoten bladeren werd snel kleiner. Aanvankelijk werden de gisten door carbendazim (A) onderdrukt. Geleidelijk bestond de gistpopulatie echter uit carbendazim-resistente stammen, zodat de carbendazim bespuitingen nauwelijks meer effect hadden. Hierdoor werden grote verschillen in gistdichtheden niet langdurig gerealiseerd. Het effect van de behandelingen op de natuurlijke infectie doorMycosphaerella graminicola (anam.Septoria tritici), Puccina recondita f. sp.tritici en op het totale dode bladoppervlak (D-waarde) wordt besproken.     

植物病害生物防治与微生态学

在概述植物病害生物防治生态学意义的基础上,从植物微生态学的角度阐述了植物根围菌及内生菌作为生防菌的依据和意义。特别是内生菌是植物体内的正常菌群,对植物病害生物防治具有重要的生态学实践意义。     

皂荚提取物对植物病原菌的抑制作用

 豆科植物皂荚是一种传统药用植物,为我国特有种。本文在室内测定了皂荚叶和棘刺乙醇提取物对枯草芽孢杆菌、胡萝卜软腐欧文氏杆菌、番茄疮痂病菌、番茄灰霉病菌、番茄叶霉病菌、棉花枯萎病菌、瓜果腐霉的抑制作用。叶提取物未检测出抗菌活性。棘刺提取物对枯草芽孢杆菌和番茄疮痂病菌都表现出一定的抗菌活性,最低抑菌浓度(MIC)分别为2.5 mg/mL和10 mg/mL。棘刺乙醇提取物在浓度为1 mg/mL时,对4种植物病原真菌的生长都表现出抑制活性,其中对瓜果腐霉生长的抑制率为28.4%。将棘刺乙醇提取物分为石油醚等5个极性不同的萃取部分,其中乙酸乙酯部分对枯草芽孢杆菌、番茄疮痂病菌、欧文氏杆菌都具有明显的抑制作用,且活性强于其它的极性段,对枯草芽孢杆菌和番茄疮痂病菌的MIC分别为1.25 mg/mL和5 mg/mL。乙酸乙酯部分对瓜果腐霉和棉花枯萎病菌的生长也有较强的抑制作用,在1mg/mL时抑制率分别为45.8%和23.9%。研究结果表明,皂英抗菌活性成分主要存在于棘刺中的乙酸乙酯极性段和正丁醇极性段中。     

Fusarium confers protection against several mycelial pathogens of pepper plants

Inoculation of nonhost pepper ( Capsicum annuum ) plants with the tomato wilt pathogen, Fusarium oxysporum f.sp. lycopersici (FOL), caused no symptoms and the fungus was not recovered from any part of the plant. FOL, however, partially protected pepper plants from subsequent infection with Phytophthora capsici , Verticillium dahliae or Botrytis cinerea by significantly reducing the percentage of diseased plants and the appearance and intensity of symptoms. FOL did not inhibit the mycelial growth of these pathogens in vitro . The protection induced by FOL against Botrytis was inhibited by 1-methylcyclopropene (MCP), an inhibitor of ethylene perception, suggesting the involvement of this hormone in the signalling of FOL-induced resistance. The activities of β-1,3-glucanase and peroxidase 48 h after FOL induction were similar to those in control plants. Chitinase activity, however, was higher in the stems of plants inoculated with FOL. A study of the levels of phenolic compounds revealed that cell-wall-bound phenolics were more abundant in plants treated with FOL, especially in stems, while soluble phenolic contents did not differ.     

玉米纹枯病拮抗内生细菌的筛选

从玉米成株期根、茎组织中分离获得232株内生细菌,在离体条件下筛选获得20株对玉米纹枯病菌具有显著拮抗作用的内生细菌,其中7株为枯草芽孢杆菌,占拮抗菌株的35%.B20-120、B20-006、B20-122、B21-072、B21-016、B20-070等6株拮抗细菌的发酵液对玉米种子萌发没有影响,进一步对其内生性(以抗利福平120 mg·mL-1为标记)及其对玉米的促生作用和玉米纹枯病的防治效果进行了研究.结果表明,供试6个拮抗菌株对玉米生长没有抑制作用,有的甚至有促进作用,并且能在体内繁殖,具可转移性.拮抗菌株B20-006和B20-120对纹枯病的防治效果最好,菌液浸种处理苗期防治效果分别为67.9%和62.3%,成株期分别为40.2%和39.1%.     

茄科植物内生细菌的分离及拮抗菌的筛选

从番茄和辣椒植株中共分离到9株对植物病原真菌具有拈抗作用的内生细菌。其中，来自樱桃番茄的内生细菌株系BS-1和湘研辣椒的内生细菌株系BS-4对辣椒根腐病菌等具有较强的拈抗作用，两者在植株体内均可以定殖。BS-1和BS-4均属于芽孢杆菌。     

Isolation of Pathogenicity Mutants of Fusarium oxysporum f. sp. melonis by Insertional Mutagenesis

Restriction enzyme-mediated integration (REMI) mutagenesis was used to isolate mutants of Fusarium oxysporum f. sp. melonis impaired in pathogenicity. The race 2 strain Mel02010 was transformed with linearized pSH75, conferring resistance to hygromycin B, with or without the enzyme used to linearize the plasmid. Addition of restriction enzymes did not affect the transformation frequency. A total of 2929 REMI transformants were tested for pathogenicity to three melon cultivars, Amus, Ogon 9 and Ohi. The race 2 strains are pathogenic to Amus and Ogon 9, but not to Ohi. Of 43 transformants with reduced pathogenicity on susceptible melon cultivars, 12 mutants were examined in detail for pathogenicity, vegetative growth and integrative mode of pSH75. The levels of pathogenicity varied among these mutants. Two mutants (B48 and B137) almost completely lost pathogenicity to both susceptible cultivars, and the others had reduced pathogenicity. Mutants B48, B241, B886 and X36 were also impaired in vegetative growth. Mutant B809 was a biotin auxotroph. By DNA gel blot analysis, nine mutants were found to contain a single copy of the transformation vector. These mutants may thus be useful in isolating genes involved in pathogenicity. Received 22 December 2000/ Accepted in revised form 16 April 2001     

Studies on amoebae and cysts associated with the isolation of Spongospora subterranea f.sp. subterranea in vitro

New evidence is presented to support the contention that the amoeba/cyst colonies isolated from surface-sterilized Spongospora subterranea f.sp. subterranea -infected potato tubers and spore balls have a saprophytic phase but are contaminants and not S. subterranea. Amoebae isolated from infected tissues and spore balls formed colonies associated with bacteria on 1% water agar at 18°C and encysted after 5–7 days. These cysts were morphologically distinct from the resting spores of S. subterranea and were formed singly or in a layer, unlike the spore ball (cystosorus) of S. subterranea . Amoebae, cysts and mixtures of amoebae and cysts in primary, secondary and tertiary subcultures failed to infect tomato roots. PCR amplification of DNA from amoebae, cysts and spore balls using the S. subterranea -specific primer pair SsF/R generated a 434-bp product from S. subterranea spore balls only and not from amoebae or cysts. When an amoeba/cyst-specific primer pair AmF/R was designed and used for PCR amplification, a single 411-bp product was generated from DNA of amoebae and cysts, but not from DNA of S. subterranea spore balls. These results are discussed in relation to earlier reports claiming the successful isolation of S. subterranea and other plasmodiophorids in vitro .     

Extracts of killedPenicillium chrysogenum Induce resistance against Fusarium wilt of melon

Dry fungal biomass ofPenicillium chrysogenum (dry mycelium), a waste product of the pharmaceutical industry, was extracted with water and applied to the roots of melon plants before or after inoculation withFusarium oxysporum f.sp.melonis (Font). Seedlings (4–6 days after emergence) treated with either acidic dry mycelium extract (DME) or neutralized dry mycelium extract (NDME) were protected against challenge infection withFom. A single drench with 2–5% DME applied 12–72 h before inoculation provided significant control of the disease compared with water-drenched, challenged seedlings. No protection was seen in plants treated 0–6 h before inoculation or 0–48 h after inoculation. Neither DME nor NDME (0.5–5%) had any effect on fungal growthin vitro, which implied that disease controlin vivo was mediated by induced resistance. The resistance induced by DME protected melon plants not only against race 1,2, but also against the three other races of the pathogen, indicating a race-non-specific resistance againstFom. Both DME and NDME significantly increased peroxidase activity and free L-proline content in seedlings 12 h and 48 h after soil drench, respectively. Resistance to Fusarium wilt was significantly associated with elevated levels of peroxidase activity but not with free L-proline content. Thus, peroxidase might be involved in the defense mechanisms activated by DME or NDME. http://www.phytoparasitica.org posting Aug. 31, 2001.     

小麦白粉病菌闭囊壳萌发及其对三唑酮敏感基线的建立

在没有甾醇脱甲基抑制剂 (DMI)类药剂用药历史的湖北利川山区麦田采集小麦白粉病菌闭囊壳 ,带回室内萌发 ,从每样本单孢子堆中分离得到61个野生菌株。采用叶段法测得每个菌株的EC₅₀,建立起小麦白粉病菌对三唑酮的相对敏感基线EC₅₀=(0.1090±0.0048)μg/ml。