花魔芋软腐病原真菌分离鉴定

魔芋软腐病是魔芋生产过程中的重要病害,也是限制魔芋产业发展的主要因素。目前,已有报道魔芋软腐病主要由细菌引起,鲜有真菌引起魔芋球茎软腐发病的报道。为明确云南曲靖市花魔芋(Amorphophallus konjac)软腐病的病原种类和侵染特征,该研究通过组织分离法,对采集自云南曲靖市的花魔芋病样进行了真菌的分离,通过形态学结合基于ITS与LSU序列分析的分子鉴定方法对分离真菌进行鉴定,并根据柯赫氏法则进行致病性测定,并对鉴定出的病原真菌同魔芋软腐病原细菌进行了双回接试验分析。结果表明:(1)从形态学和分子水平鉴定了轮纹镰刀菌(Fusarium concentricum)、尖孢镰刀菌(F. oxysporum)和F. ambrosium 3种镰刀菌,1种毛霉属真菌(Mucor sp.),1种根霉属真菌(Rhizopus sp.),1种青霉属真菌(Penicillium sp.)和1种粉红螺旋聚孢霉属真菌(Clonostachys sp.)。(2)统计分析发现,轮纹镰刀菌的相对丰度最高,为45.45%。(3)柯赫氏法则检测发现轮纹镰刀菌具有致病性。(4)轮纹镰刀菌和病原细菌胡萝卜果胶杆菌(Pectobacterium aroidearum)双接种魔芋球茎发现软腐病发病更快,病变组织重量显著高于单接种轮纹镰刀菌或果胶杆菌处理。综上表明,魔芋软腐病可能是由真菌和细菌复合侵染引发。该研究结果为魔芋软腐病的防治提供了理论依据。     

Fungi associated with storage rots of cocoyams (Colocasia spp.) in Nsukka,Nigeria

Cocoyam (Colocasia spp.) corms and cormels showing spoilage symptoms were collected from many stores in Nsukka locality and examined for rot and associated fungal pathogens. Aspergillus niger, Botryodiplodia theobromae, Corticium rolfsii, Geotrichum candidum, Fusarium oxysporum, and F. solani were recovered from rotten cocoyams. The representative isolates of these species caused cocoyam rot in pathogenicity tests. The rot due to A. niger, B. theobromae and C. rolfsii was extensive resulting in complete maceration of cocoyam tissue. Potassium sorbate (0.1 mg/ml) protected cocoyams from fungal rot with the exception of C. rolfsii.     

云南两个花魔芋主产区软腐病害植株腐烂球茎中的菌群组成及特征

【目的】软腐病是侵染魔芋的主要病害,其危害性严重,广泛传播会导致花魔芋绝产,尚无有效防控措施。魔芋软腐病的发生及暴发性传播与病原菌及其菌群有较大相关性。本研究旨在明确云南2个主产区的软腐病害花魔芋球茎及其根际土壤中的主要致病菌和优势微生物种类,分析其菌群结构特征,从而为花魔芋软腐病害的防控提供支撑。【方法】研究采集云南富源和永平2个产区的花魔芋软腐病样品,应用Illumina NovaSeq 6000测序平台进行微生物宏基因组测序和分析。同时采用选择性培养基、多级纯化培养技术以及电镜超微形态解析,分别对病害腐烂球茎中的致病菌及优势菌类进行分离鉴定和观察验证。【结果】两个花魔芋主产区软腐病害植株球茎及土壤中的微生物都非常丰富,共检出107门2 502属15 721种微生物。这2个产区的花魔芋软腐病害主要致病菌均为胡萝卜软腐坚固杆菌(Pectobacterium carotovorum),此病原菌与嗜麦芽寡养单胞菌(Stenotrophomonas maltophilia)的优势生长是2个产区软腐病害植株腐烂球茎中菌群的主要特征。此外,同产区的病害组织与土壤样品之间的菌群组成差异较大,但这2类样品中的优势菌群组成的区域性差异较小。【结论】两个花魔芋主产区软腐病害球茎组织中菌群与该区土壤菌群的相关性较低,土壤菌群区域性差异比相应的病害组织菌群区域性差异要大。因此,主要病原菌和共生菌的优势生长突破了产地差异影响,成为了病害组织菌群的主要特征,使得2个主产区软腐病害花魔芋球茎中的微生物生态系统具有高度相似性。     

Molecular detection of Pectobacterium species causing soft rot of <Emphasis Type="Italic">Amorphophallus konjac</Emphasis>

Soft rot disease of Amorphophallus konjac is caused by Pectobacterium species. Infected corms are considered a primary and important source of inocula. Based on the 16S rDNA sequences of the soft rot pathogens, one pair of specific primers was designed to identify the soft rot disease by real-time PCR and the other two were used to identify the pathogens of Pectobacterium carotovorum subsp. carotovorum. and P. chrysanthemi respectively. According to the results, a single cell of Pectobacterium could be detected by real-time PCR with the designed primer pair, while at least 100 bacteria were required for conventional PCR. Moreover, the two special primers can directly and accurately authenticate to Pectobacterium carotovorum subsp. carotovorum and P. chrysanthemi by the conventional PCR system without testing the pathogenicity, biochemical and phenotypic characterizations and so on. In conclusion, the PCR-based techniques showed several significant advantages in identifying the soft rot pathogens from konjac, such as higher sensitivity, rapidness and precision, and it could be widely used in seed quarantine.     

Leaf Spot of Alternanthera sessilis and Its Implications for Food Crops

A leaf spot of Alternanthera sessilis caused by Fusarium pallidoroseum (syn. F. semitectum) is described. During pathogenicity tests, the fungus caused leaf spots of Amarantbus sp. Abelmoschus esculentus (okra), Celosia sp. and Sorghum bicolor. It also caused rots of tubers of Dioscorea rotundata, D. alata, Solanum tuberosum, corms of Colocasia esculenta, bulbs of Allium cepa and roots of Daucus carota when wounded. Unwounded tubers, corms, bulbs and roots of these named plants were not infected except roots of Daucus carota which disintegrated between 4 and 6 days after inoculation.     

An insight into spore dispersal of <Emphasis Type="Italic">Ganoderma boninense</Emphasis> on oil palm

The disease of oil palm caused by Ganoderma boninense, although universally referred to as Ganoderma basal stem rot, occurs in three very distinct phases, with basal stem rot only part of the disease cycle. G. boninense also causes a seedling disease and an upper stem rot. An understanding of spore dispersal provides an insight into where spores of G. boninense have a role in the infection process. This role will be discussed in relation to each of these three infection phases. This understanding is a critical component of developing a successful disease control strategy.     

Reactions of six cultivars of rice (Oryza sativa L.) to three fungal diseases

Abstract

Fungal diseases are the most important threat for rice production in the world. Rice fields were visited and sampled from diseased bushes to identify fungal diseases in Kohgiluyeh and Boyer-Ahmad province, in the south-west of Iran. Fungi were isolated from diseased tissues by Agar-plate method, and purified by single-spore or hyphal-tip methods. They were identified by studying morphological characteristics. The virulence of isolated fungi was evaluated on six dominant rice cultivars, Champa of temperate region, Champa of subtropical region, Gerdeh, Shamim, Fajr and Tarom, under greenhouse conditions. The experiment was set up in a factorial experiment in completely randomized design with four replicates. The incidence of three fungal diseases: brown spot caused by Curvularia australiensis and Bipolaris cookei, foot rot caused by Fusarium globosum and Fusarium sambucinum, and black root rot caused by Exserohilum pedicellatum, was proven in this study. Although, all of the tested cultivars significantly showed different reaction to these fungi, but the cultivars Champa of subtropical region and Tarom were resistant to foot rot as well as the black root rot and the cultivar Gerdeh was resistant to brown spot disease.     

Pathogenicity of Fusarium oxysporum to Easter lily, narcissus and gladiolus

Isolates of Fusarium oxysporum from roots, bulbs and stems of Easter lilies (Lilium longiflorum) differed widely in pathogenicity and also, apparently, in tissue specificity. Virulent isolates caused a typical basal rot and root rot (but not a wilt) in which the mycelium advanced intercellularly through the scales and basal plates. Mildly pathogenic isolates became established in mature or senescent outer scales, at first producing only superficial effects, but further growth of mycelium occurred as the outer scales died and sometimes continued until the dead tissues were permeated and chlamydospores were formed. The underlying scales were then colonized. The modes of pathogenicity and survival in Easter lily were compared with those of the F. oxysporum formae causing bulb rots of gladiolus and narcissus. It is suggested that advance of hyphae by penetration between the cells of the vascular parenchyma, which is common in isolates causing rots in bulbs and corms, represents a stage in the evolution of the truly vascular habit among fusaria.     

Storage rots of taro (Colocasia esculenta) in the British Solomon Islands

Several fungicides chosen for their ability to control the pathogens previously isolated from stored corms of taro failed to prevent severe rotting. This result led to a reappraisal of the organisms involved in the initial stages of decay. Isolations made from stored corms during the first 5 days showed that Phytophthora colocasiae and Pythium splendens were the dominant fungi in the rots. Later Botryodiplodia theobromae rapidly colonized the corms to complete the decay. Attempts to reduce losses by leaving petiole bases, cormels and roots attached only succeeded in delaying infection by a few days. Corms placed in soil in well-drained pits stored relatively well up to 4 wk without impaired taste. Fungal rots were completely eliminated in corms stored in the soil but bacterial rots caused by Erwinia chrysanthemi were responsible for some decay.     

Pectolytic Bacteria Associated with Soft Rot of Calla Lily (Zantedeschia spp.) Tubers

Soft rot is the most important disease on calla lily in Poland. The isolation of the presumptive pathogen from symptomatic tubers on nutrient agar yielded bacteria with different colony morphology. Of 41 isolates collected, 10 showed pectolytic activity on crystal violet pectate medium and caused soft rot on potato slices. All pectolytic bacteria appeared to be Gram‐negative rods producing typical soft rot on inoculated leaf petioles of calla lily. Bacteria with colonies which morphologically resembled those used for inoculation were re‐isolated from diseased petioles. Their identification was based on phenotypic characters and sequence of the gene fragment coding 16S rRNA. It was found that, in addition to Pectobacterium carotovorum subsp. carotovorum, soft rot of calla lily can be caused by Pectobacterium carotovorum subsp. atrosepticum, Pseudomonas marginalis, Pseudomonas veronii and Chryseobacterium indologenes. The latter two are described for the first time as plant pathogens. The pectolytic activity of all identified bacteria, except that of P. carotovorum subsp. atrosepticum, was lower than that of P. carotovorum subsp. carotovorum, but strains of P. veronii showed a higher activity than P. marginalisand C. indologenes species.     

The involvement of ethylene in liatris corm dormancy

Corms of liatris (L. spicata, cv. Callilepsis) show a seasonal dormancy, being most active in the November harvest and least active in June. Storage of dormant corms at 3 °C for about 9 weeks resulted in a complete break of dormancy. This was accompanied by a sharp temporal increase in their rate of ethylene production, which was more pronounced in the buds than in the parenchyma tissue. Application of ethylene to the corms in the form of ethrel solution increased both ethylene production rate and sprouting. The ethylene-forming activity from ACC, measured both in vivo and in vitro, was higher in corms producing more ethylene. However, the content of 1-aminocyclopropane-1-carboxylic acid (ACC) of the corms was inversely related to their ethylene production rate. Ethylene thus seems to be involved in the dormancy control of liatris corms, and its production is apparently regulated mainly by the activity of the membranous ethylene-forming system.     

Black Rot in Zea mays Caused by Olpitrichum tenellum in China

Since 1997, severe epidemics of a black rot disease of maize (Zea mays) have occurred at Yuanmou County, Yunnan Province, China. Olpitrichum tenellum, identified by morphology of conidia and conidiophores was recovered from infected plant material. Its strong infectivity on maize and causal role in inducing black rot was confirmed by pathogenicity tests in the greenhouse. This is the first report of black rot of maize caused by O. tenellum in China.     

Disease development of Pythium root rot in bulbous Iris and Crocus

Iris bulbs and Crocus corms were planted at two planting dates in sandy soil infested with Pythium spp. At monthly intervals during the growing season root rot infection was assessed over 3 consecutive years and disease development curves were predicted for both crops. The disease development was remarkably different for Iris and Crocus and the curve shape was determined by the crop rather than by the Pythium species. Planting date had a significant effect on disease development in both crops. No correlation was found between disease development and soil temperature.     

Antifungal activity of some plant extracts on some pathogenic fungi

The inhibitory activity of five plant extracts viz. Artemisia absinthium L., Rumex obtusifolius L., Taraxacum officinale Weber ex Wiggers, Plantago lanceolata L. and Malva sylvestris L. were evaluated against the mycelial growth of three fungi Alternaria alternata (Fr.) Keissler, Penicillium expansum Link ex Thom. and Mucor piriformis Fisher that cause rot diseases in fruits and vegetables resulting in low yield and quality of fruits and vegetables. Results revealed that all the concentrations of plant extracts brought about significant inhibition in the mycelial growth of these pathogenic fungi. However, the highest concentration caused maximum inhibition in the mycelial growth followed by lower concentrations of plant extracts. The extract of A. absinthium leaves at highest concentration (S) proved highly effective in inhibiting the mycelial growth of all these pathogenic fungi followed by other plant extracts. These plants thus may have potential as the new natural fungicide for management of fungal rot diseases.     

Characterization of blackleg and soft rot from potato in northern Thailand

Blackleg and soft rot of potato cause economic loss through reduced yield and quality. The causal agents of bacterial blackleg and soft rot of potato were identified based on biological data and sequence analyses of the 16S rDNA gene. Between 2016 and 2018, diseased potato stems and tubers were collected in Chai Prakan District, Chiang Mai Province, and Chiang Khum District, Pa Yao Province. The symptoms included black stem lesions, soft rot on tubers, wilting, break down of the stem vascular ring and foliar yellowing. Of 13 bacterial isolates, five were identified as Pectobacterium carotovorum subsp. brasiliense, four‐Dickeya dadantii, two‐Pseudomonas putida and two‐Bacillus altitudinis. Pathogenicity tests of P. carotovorum subsp. brasiliense and D. dadantii resulted in lower leaves turning yellow and wilting followed by blackleg symptoms on lower stems and maceration of tuber tissue. Symptoms caused by P. putida were yellowing and wilting of leaves. B. altitudinis caused yellowing of the lower leaves and wilting followed by drying of leaf tissue. This is a first report of these bacterial pathogens causing blackleg and soft rot of potato in Thailand.     

Comparison of methods to evaluate the potential of fungal growth on decay of spruce wood after short-time treatment

Several analytical methods were compared to evaluate characteristic wood decaying fungi for their potential to depolymerise lignin on spruce wood particles. Wood samples were treated with the white rot fungi Phlebia brevispora, Ceriporiopsis subvermispora, Merulius tremellosus, Pycnoporus sanguineus, Trametes pubescens and with the brown rot fungus Gloeophyllum trabeum. The UV absorbancies of crude ethanol extracts, total extractives content from sequential extraction, ligninolytic enzyme activities, lignin solubilisation and decrease of lignin content were compared. It was shown, that, in early decay stages, UV absorbancies of crude ethanol extracts and total extractives content correlate well with lignin degradation, increase of acid soluble lignin and increased production of ligninolytic enzymes (total peroxidase). Lignin content was determined using FT-NIR spectroscopy as well as by wet-chemical analysis, indicating a very good correlation between the two methods. According to the different analytical methods, the tested fungi can be classified into three categories based on their characteristic behaviour: brown rot, “slow” and “fast” white rot.     

Differential selection pressure exerted by root rot disease on the microbial communities in the rhizosphere of avocado (Persea americana Mill.)

Despite the high economic impact of root rot disease, knowledge regarding the rhizosphere microflora of avocado trees affected by root rot is limited. Metagenomics was applied to identify the difference in the rhizosphere microflora of avocado trees with and without visible symptoms of root rot. Approximately, 446,970 common gene catalogues differed between them, confirming that root rot affected the bacterial and fungal communities in the rhizosphere. The proportion of bacterial genera, namely, Labilithrix, Sorangium, Sandaracinus and Pedosphaera showed a decrease while Phenylobacterium, Rhizomicrobium, Candidatus Solibacter and Silvibacterium genera were increased by root rot. The proportion of fungal genera, namely, Pseudogymnoascus, Moelleriella, Mortierella, Lepidopterella, Babjeviella, Lachancea, Macrophomina, Pneumocystis, Sugiyamaella and Cyphellophora showed an increase while Cryptococcus, Verticillium, Bipolaris, Pyrenochaeta, Rhizophagus, Cenococcum and Neonectria genera were inhibited by root rot. Moreover, the proportion of the top 10 bacteria in the rhizosphere of symptomatic trees was significantly higher, and that of the top 10 fungi was significantly lower, compared to the asymptomatic trees. Principal component analysis based on abundance analysis and function prediction showed that in symptomatic trees, the bacterial community was more concentrated, while the fungal community was more dispersed. The differences in the responses of bacterial and fungal genera suggested that the pathogenic fungi exert varying selection pressure on the microflora. Moreover, root rot affected the metabolism of carbohydrates, lipids and amino acids in bacteria, and the global and overview maps, carbon metabolism and processing of genetic and environmental information in fungi, which might result in differential selection pressure.     

Identification of Colletotrichum Species Causing Bitter Rot of Apple and Pear in Croatia

Symptoms of bitter rot were observed on apple and pear fruits in the field and in storage in Croatia between 2009 and 2011. Fifteen Colletotrichum isolates from apple and two from pear were collected and identified by sequencing of the ITS1 and ITS2 regions of the ribosomal DNA. Phylogenetic analysis revealed that ten isolates from apple and two isolates from pear could be identified as Colletotrichum fioriniae, five isolates from apple were clustered in Colletotrichum clavatum, while one isolate was in the Colletotrichum acutatum A7 group. All isolates caused typical bitter rot symptoms when inoculated on apple and pear fruits.     

Endangered Cactus Restoration: Mitigating the Non-Target Effects of a Biological Control Agent (Cactoblastis cactorum) in Florida

The moth Cactoblastis cactorum (Berg), the poster child of weed biological control in Australia, has recently invaded the United States and threatens native cacti. Concern is greatest for the endangered semaphore cactus, Opuntia corallicola, of which only two known populations exist in the wild. We made three separate outplantings of O. corallicola, designed to bolster the number of extant cacti and to test the effectiveness of three different treatments to protect the cacti from Cactoblastis. In one outplanting, we tested the associational susceptibility hypothesis and found that cacti planted more than 20 m away from the common prickly pear cactus, Opuntia stricta, which act as a reservoir of Cactoblastis, were just as frequently attacked and killed by Cactoblastis as cacti planted within 5 m. In addition, Cactoblastis attack was greater in the shade than in the sun. In the second outplanting, we minimized the attack from Cactoblastis by using protective cages planted at least 500 m from O. stricta in areas not inhabited by cacti. Cages attracted the attention of local animals, which destroyed the cages and trampled the cacti inside to death. Crown rot caused high mortality in this outplanting. In the third outplanting, again conducted at least 500 m away from O. stricta, fertilization did not reduce crown rot mortality. We suggest that increasing populations of O. corallicola in Florida, by means of outplantings, will remain a challenge because of death from Cactoblastis when planted in areas where cacti normally grow and because of death from crown rot in areas where they do not. Because Cactoblastis is moving rapidly northward and westward and has already reached Charleston, South Carolina, rare cacti in the rest of the U.S. Southeast may be in danger. Eventually, many cactus species in the U.S. South, Southwest, and Mexico will likely be threatened by this moth.