马铃薯立枯丝核菌拮抗菌QHZ11的实时荧光定量PCR快速检测与应用

【背景】植物根际促生菌(plant growth-promoting rhizobacteria,PGPR)在根际的定殖是其发挥作用的基础,直观有效的跟踪技术和定量方法是研究PGPR在根际原位分布规律的重要工具。【目的】建立一种马铃薯黑痣病病原菌——立枯丝核菌拮抗菌QHZ11的实时荧光定量PCR快速检测体系,并检测拮抗菌QHZ11在马铃薯根际的动态变化。【方法】根据GenBank中登录的类芽孢杆菌及近源菌株gyrB基因序列差异筛选特异性引物,优化反应条件;通过盆栽试验对马铃薯根际拮抗菌进行快速检测。盆栽试验设3个处理,T1:对照(无菌水,CK);T2:QHZ11菌悬液灌土(QHZ11);T3:将功能菌在有机肥中进行二次固体发酵制成生物有机肥(BOF11)。【结果】筛选出拮抗菌QHZ11的专用引物为gyrB-F/gyrB-R;建立的拮抗菌QHZ11实时荧光定量PCR检测方法特异性好、灵敏度高且重复性较好,线性相关系数为0.999 8,检测组内变异系数均在1%以内,扩增效率为0.9,可检测出1×103-1×1010copies/g-soil的拮抗菌,具有检出限低和扩增效率高的特点。盆栽试验...     

In vitro and greenhouse evaluations of Trichoderma isolates for biological control of potato wilt disease (Fusarium solani)

The biological efficacy of Trichoderma species may differ due to variations in ecosystems. This study was conducted to assess the biocontrol efficacy of some native Trichoderma isolates against Fusarium solani, an important causal agent of potato wilt disease under laboratory and greenhouse conditions at Shahrood Agricultural Research Centre, Shahrood, Iran, during 2006–2007. Fourteen isolates were collected among which eight showed promising ability in inhibiting the growth of the pathogen through dual culture and production of volatile and non-volatile inhibitors but T. brevicompactum (T1), T. longibrachiatum (T5) and T. asperellum (T2) were almost better than other isolates in inhibiting the mycelial growth of the pathogen in comparison to control in the above three tests (p ≤ 0.01). Isolates performing mycoparasitism under in vitro condition were evaluated against the disease in pot culture under greenhouse condition. In all treatments in which Trichoderma isolates + F. solani were involved lower disease incidence was noticed in comparison to Fusarium-infested control (p ≤ 0.05). Best disease control was observed in potted plants treated with F. solani + T. longibrachiatum (T5) with 6.25% disease incidence in comparison to Fusarium-infested control, in which disease incidence was observed to be 75%. Interaction of T. brevicompactum (T1) and F. solani also indicated good control of the disease by 12.50% of disease incidence.     

Studies on inhibitory activities of antibodies against phytopathogenic fungi: an immunological approach

The major problem in agriculture is that of the fungal pathogens. In this era, where biological control is at focus, and is the centre of crop protection as well as environmental protection, the synthesis of new bio bodies is of utmost need. Fungicides available in the market, though of potential, are pathogen specific and highly pollutive. An attempt was made to raise polyclonal antibodies against Aspergillus niger. Following the particular standardised immunisation schedule, regular injections and periodic tapping were carried out. The IgG purified was used to check cross-reactivity with different crop fungi like Fusarium solani, Fusarium oxysporum f. sp. cicer (FOC), Rhizoctonia bataticola, Botryodiplodia theobromae, Curvularia sp., Alternaria porri and also with Aspergillus niger by two different methods. Liquid test media and the radial growth inhibition test performed in solid media were used to check the inhibition of fungi and cross-reactivity. The results were subjected to analysis of variance (ANOVA) by using Tukey's test at the significance level of p < 0.05. The antibodies were active against all the fungi for more than 15 days except for A. niger in which from the seventh day onwards spore germination was observed. The probable role of antibodies to detect the common antigenic molecule that may be present in all the tested fungi and their role in inhibiting these fungi is discussed.     

A synergism between oxalic acid and polygalacturonases in the depolymerization of potato tuber tissue

Rhizoctonia bataticola produced oxalic acid in vitro and in vivo during pathogenesis of patato tuber. Polygalacturonase (PG) was also detected in culture filtrates of the rot-causing organism. Levels of maceration and cell death in tuber tissue were higher when a mixture of oxalic acid and PG was used than when either oxalic acid or PG were used alone.     

Epidemics of Rhizoctonia Root Rot in Association with Biological and Physicochemical Properties of Field Soil in Bean Crops

During two growing seasons (2008 and 2009), the associations of Rhizoctonia root rot (RRR) with a number of soil properties were determined at different growth stages in 122 commercial bean fields in Zanjan, Iran. Mean RRR incidence at a level of 4–25% sand content was lower than that at 45–65% level. Damage by fly puparia had no significant effect on RRR incidence and occurrence. A greater RRR incidence was detected in field soils treated with fungicides compared with non‐treated soils. A lower RRR incidence was associated with the highest level of soil organic matter (1.2–1.8) compared with the lowest level, 0.4–0.8. The highest RRR incidence corresponded with no rhizobial nodulation compared with highly nodulated bean roots. RRR incidence was negatively correlated with soil silt and organic matter content at R6–7 and R9 growth stages. RRR‐affected fields were recognized with a greater soil pH (V3) and sand content (R9), and a lower silt (R9) and organic matter content (R6–7 and R9) in comparison with RRR‐free fields. Loadings and linear regressions between RRR incidence and principal component scores indicated that the most effective soil characteristic linked to the disease was silt at V3, sand at R6–7 and organic matter at R9 stage. This new epidemiological information extends our knowledge of the bean–RRR–soil interaction on a regional basis.     

A comparison ofin vitro andin vivo effect of clay minerals,humic acid and micronutrients on the activity of fungicides againstRhizoctonia solani

Activity of nine fungicides against mycelial growth ofRhizoctonia solani in potato dextrose broth and in pot tests as seed treatment against cowpea seedling rot in infested soil was differentially in fluenced by clay minerals, humic acid and micronutrients. Humic acid, extracted from farmyard manure, considerably lowered the activity, bothin vitro andin vivo, of all fungicides except chloroneb. Montmorillonite caused substantial decrease in disease control by fungicides but enhanced the toxicity of 2-methoxyethyl mercury chloride (MEMC), and quintozene in culture. Kaolinite inactivated carbendazim, benomyl and thiophanatemethyl in nutrient broth but had little effect on disease control by these fugicides. The six micronutrients altered the activity of fungicides to varying levels often without definite correlation betweenin vitro andin vivo results. The results ofin vitro growth inhibition tests are largely inapplicable to dieseas control tests in infested soil mainly due to the differences in the ambient conditions of the two systems.     

Mycorrhizal differences between genuine roots and tuberous roots of adult plants ofSpiranthes sinensis var.amoena (Orchidaceae)

Genuine roots ofSpiranthes sinensis var.amoena were infected with the mycorrhizal fungusRhizoctonia repens immediately after root formation in autumn. Infection by the mycorrhizal fungus extended, reaching a maximum the following early summer. The amount of living mycorrhizal fungus in the genuine roots dramatically declined in the flowering season, and then the roots decomposed. Tuberous roots were formed in spring. Mycorrhizas were limited to local infections and did not spread along the roots. The infection level of living mycorrhizal fungus in the tuberous roots was less than in the genuine roots throughout the year. The amount of dead fungal coils in the tuberous roots increased as the tuberous roots aged. The mycorrhizal characteritics of tuberous roots ofS. sinensis var.amoena were totally different from those of genuine roots although the tuberous roots morphologically resembled the genuine roots. Contribution No. 96, Laboratories of Plant Pathology and Mycology, Institute of Agriculture and Forestry, University of Tsukuba.     

Use of DAPI for Anastomosis Group Typing of Strains of the Fungus Rhizoctonia solani

Strains of Rhizoctonia solani, a common soil-borne, pathogenic fungus of plants, are assigned to one of 11 anastomosis groups (AGs) based on the occurrence of imperfect fusions (anastomoses) between hyphae of a non-typed strain and a tester strain of one of the 11 AG's. Imperfect fusion is characterized by the death of one or more cells in each of the hyphae involved in the fusion. Although hyphae from branches of the same strain of JR. solani may fuse with each other (self-fusion), cell death does not occur. Cell death is accompanied by nuclear degradation and granulation, or plasmolysis of the cytoplasm, which often is not visible using bright-field microscopy. When the DNA-binding fluorochrome DAPI (4', 6-diamidino-2-phenylindole) is used and the hyphal fusions viewed under fluorescence microscopy, no nuclei are observed in fused hyphal cells from two strains of the same AG of R. solani Because DAPI reacts only with living nuclei, lack of staining is presumptive evidence that the fused cells are dead as a result of imperfect fusion. The use of DAPI reduces the time required for making AG determinations compared to standard methods because it eliminates the need to assess cell wall dissolution and cytoplasmic fusion. Also, it is not necessary to trace the hyphae involved in the fusion to their respective origins to ensure that self-fusion has not occurred.     

喀麦隆芋艿根腐病病原菌鉴定

从采自喀麦隆不同地区的芋艿(Xanthosoma mafaffa L.)病根及田土中分离菌株,分别从雅温得(Yaound(?))和巴马约(Mbalmayo)分离到的腐霉菌株XPMY和XPMM1,根据其形态学及生理学特征鉴定为群结腐霉Pythium myriotylum,用上述腐霉菌株的游动孢子悬液或菌丝体片断悬液人工接种,表现叶黄化及根腐等典型症状。用经常伴随群结腐霉的立枯丝核菌Rhizoctonia solani及茄镰孢Fusarium solani人工接种后均未表现症状。研究结果表明:群结腐霉Pythium myriotylum Drechsler是喀麦隆芋艿根腐病的致病菌。