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.     

Effect of peat-bran inoculum ofTrichoderma species on biological control ofRhizoctonia solani in lettuce

A range of known biocontrol or plant growth-stimulating species ofTrichoderma orGliocladium were grown on peat-bran substrate to yield between 5×10⁷–3×10¹⁰ colony forming units (cfu's)g^–1 substrate after 14 days growth. Inocula were incorporated into peat:sand potting compost infested withRhizoctonia solani to give 7–8 × 10⁴ cfu's of antagonist g^–1 compost and assessed for biological control activity using lettuce seedlings. Six of the eight antagonists decreased daming-off and three of these consistently increased yield in comparison withR. solani treatment alone.Subsequently, peat-bran inoculum ofT. harzianum isolate TH1 was incorporated at 0.5% w/v intoR. solani infested potting compost. Both autoclaved and nonautoclaved inoculum ofT. harzianum TH1 decreased disease and increased yield. Incorporation of ethyl acetate-extracted autoclaved inoculum ofT. harzianum TH1 resulted in similar levels of biocontrol and improved plant growth as did incorporation of nonautoclaved and autoclavedT. harzianum TH1 inoculum. The need to standardize inocula and controls is emphasized.     

Induction of cytoplasmic leakage from Rhizoctonia solani hyphae by Gliocladium virens and partial characterization of a leakage factor

The biocontrol fungus Gliocladium virens (Gl‐21)was grown on various solid and liquid substrates. Aqueous extracts of wheat bran and peanut hull meal (PHM)as well as spent glucose tartrate broth (GTB), Czapek‐Dox broth (CDB) and potato dextrose broth (PDB) upon which Gl‐21 was grown caused leakage of carbohydrates and electrolytes from hyphae of the soilborne plant pathogen Rhizoctonia solani. In addition, the mycelial weight of R. solani was reduced. Most of the leakage factor was formed in the substrates within six days of incubation. Acidification of the bran and the PHM media before inoculation stimulated production of the leakage factor by G. virens. Size‐fractionation experiments indicated that a combination of factors produced by G. virens induced leakage from R. solani. Although the < 1 k Da fraction from the water‐extracted bran culture contained significant leakage activity, it was less than in the non‐fractionated bran culture. The > 1 k Da fraction from bran culture did not induce leakage activity. When the < 1 k Da and the > 1 k Da fractions were recombined, leakage activity was restored to a level similar to that of the non‐fractionated preparation. Gliotoxin was detected in culture filtrates from G. virens grown on bran and PHM media. Gliotoxin preparations induced leakage of carbohydrates and electrolytes from R. solani and caused a concomitant reduction in mycelial weight, which suggests that it is a leakage factor.     

Cowpea Root Rot Severity and Metabolic Changes in Relation to Manganese Application

Severity of root rot (Rhizoctonia solani and Rhizoctonia bataticola) of cowpea (Vigna unguiculata L.) was reduced by 42.7 and 42.0%, respectively over control following the application of 10 μg/g Mn as manganese sulphate. Reduction in disease incidence was associated with increased levels of polyphenol oxidase (PPO), peroxidase (PO) and total phenols. PO activity was several times more as compared with PPO‐specific activity and increased markedly after infection either with R. solani or R. bataticola. Contrary to PPO and PO, the specific activity of catalase declined sharply. Infection also caused an increase in the content of reducing sugars, Cu, Zn and Mn but a decrease in o‐dihydric phenols, flavanols, total soluble sugars, non‐reducing sugars and Fe contents. It is suggested that Mn at the rate of 10 μg/g soil can be used to manage the root rot of cowpea.     

Combination of Pseudomonas aeruginosa and Pochonia chlamydosporia for Control of Root-Infecting Fungi in Tomato

A plant growth‐promoting rhizobacterium, Pseudomonas aeruginosa strain IE‐6, and a fungal antagonist, Pochonia chlamydosporia, were tested for their ability to inhibit mycelial growth of root‐infecting fungi under laboratory conditions including Macrophomina phaseolina, Fusarium oxysporum, F. solani and Rhizoctonia solani. Biocontrol effectiveness of the bacterium and the fungus alone or in combination was also determined for the control of root‐infecting fungi under field conditions. In a dual‐culture plate assay, the colonies of P. chlamydosporia and P. aeruginosa met each other and no further growth of either organism occurred. Against M. phaseolina, F. solani and R. solani, an ethyl acetate extract of the culture filtrates of P. aeruginosa inhibited fungal growth greater than the hexane extract, but against F. oxysporum the hexane extract caused greater inhibition of fungal growth. By contrast, against M. phaseolina, F. oxysporum and F. solani, the hexane extract of P. chlamydosporia was more effective in the inhibition of fungal growth than the ethyl acetate fraction. Ethyl acetate extracts of P. aeruginosa at 1.0 mg/ml not only inhibited the radial colony growth of R. solani but also lysed the fungal mycelium. P. aeruginosa produced siderophores and hydrogen cyanide under laboratory conditions. Field experiments conducted in 1997 and repeated in 1998 revealed that Pochonia chlamydosporia and P. aeruginosa significantly suppressed the root‐infecting fungi M. phaseolina, F. oxysporum, F. solani and R. solani and that the combination of the two caused greater inhibition of the fungal pathogens than either alone. Application of P. chlamydosporia and P. aeruginosa as a soil drench also resulted in enhanced growth of tomato plants.     

The role of cuticle and epidermal cell wall in resistance of rapeseed and mustard to Rhizoctonia solani

A study was conducted to determine whether the cuticles in two genera of the family Cruciferae are effective barriers to infection by Rhizoctonia solani, and whether differences in cuticle and epidermal cell wall thickness and morphology of epicuticular wax exist between resistant and susceptible cultivars. As Canola/rapeseed (Brassica napus) and mustard (Sinapis alba) plants develop from 1 to 3 weeks of age, they become increasingly resistant to R. solani AG2-1 seedling root rot. Seven-day-old seedlings of S. alba cultivars are invariably more resistant than B. napus cultivars. Brassica napus cultivars do not show an obvious cuticle layer at 1 week but at 3 weeks the presence of a cuticle is seen through autofluorescence with a concomitant increase in resistance to R. solani. Removal of the cuticle from 3-week-old hypocotyls by chloroform treatment results in a decrease in cuticular autofluorescence and a significant increase in disease severity in both resistant and susceptible cultivars. Three-week-old plants of S. alba have a much lower percent disease rating and a significantly (p=0.05) thicker cuticle layer than similar-age plants of B. napus. The results suggest that the cuticle plays an important role in the resistance of S. alba and older plants of B. napus to infection by R. solani.     

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.     

Seedling mycorrhiza: a discussion of origin and evolution in Orchidaceae

Recent phylogenetic analyses confirm the monophyly of Orchidaceae as sister group to the remainder of Asparagales, and identify the sequence of early branching lineages in Orchidaceae. Orchid seedling mycorrhiza (OSM) involving rhizoctonious fungi is distributed widely in all subfamilies, including the first branching ones, and its status as a founding event is thus supported. OSM is recognized as one element in the character syndrome that distinguishes orchid biology, and we argue that OSM was the first to evolve. We also discuss the possible evolutionary origins of OSM in Asparagales. The prevalent mycobionts suggest a derivation from a pathogenic relationship, and sister group comparison offers little support for derivation from other mycorrhizal relationships. A combination of in situ sowings and molecular identification of seedling mycobionts has established that a broad range of fungi besides rhizoctonious mycelia are presently involved in OSM, presumably evolving secondarily and often in parallel in different orchid clades. Structural features and internal patterns of mycobiont behaviour appear to have remained largely the same, implying that OSM needs only minor physiological adjustment to accommodate new mycobionts. Such modifications will have involved checkpoints for recognition/rejection and the formation/breakdown of pelotons. These physiological mechanisms are so far largely unknown. The trophic versatility of the mycobionts and the apparently easy shifts could be a main factor in the ecological adaptability of orchids and proliferation of the family. © 2014 The Linnean Society of London, Botanical Journal of the Linnean Society, 2014, 175 , 313–327.     

实时定量PCR法预测禾谷丝核菌Rhizoctonia cerealis基因组大小

【目的】准确测定基因组大小是进行禾谷丝核菌Rhizoctonia cerealis全基因组序列测定和拼接的基础,本研究旨在利用实时定量PCR方法预测禾谷丝核菌的基因组大小。【方法】首先克隆了禾谷丝核菌R0301菌株翻译延伸因子A基因(tef A)的部分序列,Southern杂交明确该基因在该病菌基因组中为单拷贝。以已测序立枯丝核菌(Rhizoctonia solani)AG1-IA融合群菌株GD118为对照,采用实时定量PCR的方法进行了禾谷丝核菌基因组大小的预测。【结果】实时定量PCR的方法可以比较准确的测定立枯丝核菌基因组的大小,研究首次预测了禾谷丝核菌的基因组大小位于32.2–36.6 Mb之间。【结论】实时定量PCR法是一种快速和简便的预测丝核菌基因组大小的方法。     

Interaction of Rotylenchulus reniformis with Seedling Disease Pathogens of Cotton

The impact of 10 Fusarium species in concomitant association with Rotylenchulus reniformis on cotton seedling disease was examined under greenhouse conditions. In experiment 1, fungal treatments consisted of Fusarium chlamydosporum, F. equiseti, F. lateritium, F. moniliforme, F. oxysporum, F. oxysporum f.sp. vasinfectum, F. proliferatum, F. semitectum, F. solani, and F. sporotrichioides; Rhizoctonia solani; and Thielaviopsis basicola. The experimental design was a 2 × 14 factorial consisting of the presence or absence of R. reniformis and the 12 fungal treatments plus two controls in autoclaved field soil. In experiment 2, the same fungal and nematode treatments were examined in autoclaved or non-autoclaved soil. This experimental design was a 2 × 2 × 14 factorial consisting of field or autoclaved soil, presence or absence of R. reniformis, and the 12 fungal treatments plus two controls. In both tests, Fusarium oxysporum f. sp. vasinfectum, F. solani, R. solani, and T. basicola consistently displayed extensive root and hypocotyl necrosis that was more severe (P ≤ 0.05) in the presence of R. reniformis. Soil treatment (autoclaved vs. non-autoclaved) influenced the impact of the Fusarium species on cotton seedling disease, with disease being more severe in the autoclaved soil. Rotylenchulus reniformis reproduction on cotton seedlings was greater in field soil compared to autoclaved soil (P ≤ 0.05). This study suggests the importance of Fusarium species and R. reniformis in cotton seedling disease.