番茄青枯病内生拮抗细菌的筛选

 从广西一些市县采集番茄茎标本分离得到55个细菌菌株,分属为芽孢杆菌(Bacillus spp.)、黄单胞菌(Xanthomonas spp.)、假单胞菌(Pseudomonas spp.)和欧文氏菌(Erwinia spp.),其中芽孢杆菌为优势种群。经回接测试,有36个菌株为番茄植株内生菌。这些内生菌只有7个菌株对番茄青枯病菌有拮抗作用,芽孢杆菌B₄₇菌株对番茄青枯病菌拮抗作用较强,经室内和田间初步防治测定,它对番茄青枯病有较好的防治效果。     

Genetic diversity and structure of Hemileia vastatrix populations on Coffea spp.

Coffee leaf rust is the most limiting disease for coffee cultivation in Brazil. Despite its importance, relatively little is known about the genetic diversity of Hemileia vastatrix, the rust causal agent. In this work, the DNA from 112 monopustule isolates from different geographic locations and coffee genotypes were analysed by amplified fragment length polymorphisms (AFLP). The objectives were to assess the influence of the host and geographic origin on the diversity and population differentiation in H. vastatrix. The fungal population showed a low level of genotypic diversity. Gene diversity (h) was 0·027 and the hypothesis of random mating in the total population was rejected, but evidence for recombination was found for two subpopulations (São Paulo and Paraná). The analysis of molecular variance revealed that 90% of the genetic distribution of the pathogen occurs among isolates within the subpopulation (states or host of origin). There was no correlation between geographic and genetic distance (r = ?0·024, P = 0·74), which together with the high number of migrants and the low degree of differentiation in populations of H. vastatrix, is consistent with the fact that the inoculum is probably easily dispersed by wind over long distances, allowing dispersal of the pathogen among coffee growing areas in Brazil. Therefore, it is difficult to predict the durability of resistant sources to coffee rust. The recommendation for the breeding programmes is thus to incorporate multigenic resistance as a control strategy.     

The effect of fungicides on fungal antagonists of onion white rot and selection of dicarboximide-resistant biotypes

The sensitivity of four fungal antagonists (Chaetomium globosum, Trichoderma harzianum, T. viride, Trichoderma sp.) to six fungicides was evaluated. The fungi were insensitive to captan, mancozeb and thiram but were sensitive to benomyl (EC₅₀ < 0 3/μg/ml) and the two dicarboximides iprodione and procymidone (EC₅₀ < 3 3/μg/ml). Conidia and mycelia of the four fungi were exposed to a series of ultraviolet irradiations in order to select biotypes resistant to benomyl and iprodione. Resistance to benomyl was not induced but a number of biotypes of each fungus were isolated from irradiated cultures that showed resistance to iprodione (EC₅₀ values of up to 56·4, 177·8, 171·5 and 216·4/μg/ml for Chaetomium globosum, Trichoderma harzianum, T. viride and Trichoderma sp., respectively). With three exceptions, all selected biotypes retained their fungicide-resistance after being extensively cultured on fungicide-free medium. In general, growth of the fungicide resistant biotypes on MEA plates and in MYE liquid medium was equal to that of the corresponding wild-type strains. One resistant biotype of Trichoderma sp. (C62UV3) grew significantly (P < 0·05) better both on solid and in liquid medium. This resistant biotype was also observed to have greatly enhanced conidial production on agar plates. All fungicide resistant-biotypes retained the ability to antagonize the pathogen Sclerotium cepivorum in dual culture and, in two instances (A53UV1, A53UV5), exhibited greater antagonism as evidenced by the production of larger inhibition zones. Similarly, with two exceptions, the resistant biotypes retained the ability to control onion white rot disease caused by S. cepivorum in the glasshouse. In particular, two fungicide-resistant biotypes (Trichoderma sp. C62UV3 and Chaetomium globosum A53UV1) reduced white rot of onion more effectively than did their respective wild-type parents.     

The effect of leaf age on infection of Coffea genotypes by Hemileia vastatrix

The age of a leaf at the time of inoculation affected both development of infection structures on the leaf surface and subsequent sporulation. In the case of the susceptible genotypes, cv. Caturra and line NC-169, there was a decrease in the number of appressoria formed, with an increase in age of fully expanded leaves. On the selected resistant line, NP-547, significantly fewer appressoria were formed on the fourth leaf pair than on younger or older leaf pairs. Appressoria were formed significantly more readily over stomata of leaf pairs one, two, three, four and six than over five and seven of the susceptible selections. On the youngest leaf pair a significantly large percentage of appressoria formed away from stomata compared with the percentages obtained for older leaf pairs, where a large proportion of germinated uredospores did not form appressoria. The reaction type of the susceptible genotypes was a greater value on younger than older leaf pairs, and the reverse was true of the resistant genotype. The latent period (inoculation to sporulation) was shorter on NC-169 than on cv. Caturra. No uredospores formed on NP-547. Uredosorus density decreased with an increase in leaf age on the susceptible genotypes.     

The effect of leaf age on incomplete resistance of coffee to Hemileia vastatrix

The influence of leaf age on incomplete resistance to race II of coffee leaf rust (Hemileia vastatrix), which is the most common race in Brazil, was tested in the greenhouse and in the field. Three stages of leaf age were distinguished: young leaves which had just fully expanded and still had a shiny appearance on the day of inoculation, adult leaves of 1 to 4 months old, and old leaves of 6 to 12 months old which were formed in the foregoing growing season.The effect of leaf age varied with the coffee genotype. With the susceptibleCoffea arabica cvs Mundo Novo and Catuai no important effect of leaf age on latency period and reaction type was observed. Lesion density, depending on inoculation method, was affected to some extent, older leaves showing lower values. With genotypes of theC. canephora cv. Kouillou, which varied in level of incomplete resistance, adult leaves appeared to be more resistant than young and old leaves. This adult leaf resistance was expressed mainly by a lower lesion density, but also by a longer latency period and a lower reaction type.Within the hybrid populations Icatu and Catimor, genotypes with race-specific incomplete resistance were tested and others of unknown specificity. Resistance was expressed mainly by a low reaction type. Generally, resistance decreased with increasing leaf age, which was occasionally associated with a complete change in reaction type. Contrary to cv. Kouillou, in Icatu and Catimor no higher susceptibility of young leaves than of adult leaves was observed.The incomplete resistance to race I of the differential CIFC H152/3, heterozygous for S_H4, was also better expressed in young leaves, older ones becoming gradually more susceptible.It is concluded that for screening incomplete resistance to coffee leaf rust leaves of different ages should be tested.Samenvatting Het effect van bladleeftijd op incomplete resistentie van koffie tegen fysio II van koffieroest (Hemileia vastatrix), het algemeen voorkomende fysio in Brazilië, werd getoetst in de kas en in het veld. Drie stadia in de bladleeftijd werden onderscheiden: jong blad dat net volgroeid was en nog glansde op de dag van inoculatie, volwassen blad van 1 tot 4 maanden oud en oud blad van 6 tot 12 maanden oud dat gevormd was in het voorafgaande groeiseizoen.Het effect van bladleeftijd varieërde met het koffiegenotype. Bij de vatbareCoffea arabica rassen Mundo Novo en Catuai werd geen belangrijke invloed waargenomen van bladleeftijd op latentieperiode en reactietype. Lesiedichtheid werd, afhankelijk van de inoculatiemethode, wel enigszins beïnvloed. Bij vijf genotypen van hetC. canephora ras Kouillou, die varieerden in niveau van incomplete resistentie, bleek volwassen blad veelal resistenter dan jong en oud blad. Deze resistentie uitte zich voornamelijk door een lagere lesiedichtheid maar ook door een langere latente periode en een lager reactietype.Van de hybride populaties Icatu en Catimor werden enkele genotypen met fysiospecifieke incomplete resistentie getoets alsmede genotypen waarvan de specificiteit van de resistentie onbekend was. Verschillen in resistentie uitten zich voornamelijk door verschillen in reactietype. Over het algemeen nam de resistentie af bij toenemende bladouderdom. Anders dan bij de cv. Kouillou werd bij Icatu en Catimor geen grotere vatbaarheid van jong ten opzichte van volwassen blad waargenomen. Bij sommige genotypen kwam resistantie zelfs alleen maar tot uiting in het jonge blad, terwijl het volwassen blad normaal vatbaar was. Een soortgelijk effect van bladleeftijd werd waargenomen bij inoculatie van de koffiedifferentieel CIFC H153/3, die heterozygoot is voor het resistentiegen S_H4, met het incompatibele fysio I.Er wordt geconcludeerd dat voor het bepalen van incomplete resistentie tegen koffieroest bladeren van verschillende ouderdom getoetst dienen te worden.     

The effect of light intensity on incomplete resistance of coffee to Hemileia vastatrix

Resistance of coffee to race II ofHemileia vastatrix was tested in different environments at light intensities (LI) from 17 to 100% of total outdoor radiation. Nine treatments, in which three levels of LI before inoculation were combined with three levels of LI after inoculation, were applied to seedlings of the susceptible cv. Mundo Novo. Higher LI before inoculation induced a significant increase in lesion density, whereas the opposite was observed for treatments after inoculation. Maximum differences in lesion density were threefold. The interaction between pre-and post-inoculation treatments was also significant. Necrosis of lesions occurred under extremely high LI after inoculation.Genotypes of the Icatu population and ofCoffea canephora cv. Kouillou, which varied in disease level in the field, were tested in different environments, constant LI being applied before and after inoculation. Most genotypes were more resistant at low LI than at high LI, paralleling the results obtained for the control cv. Mundo Novo. With cv. Kouillou, sporulating lesion density, latency period and reaction type were significantly affected by LI and genotype. The interaction between LI and genotypes was significant for sporulating lesion density and reaction type, mainly because the most resistent genotype was not affected, or affected in opposite direction, by LI.Environment affected the expression of the resistance gene S_H4. Observations on a segregating F2 population indicated dominant gene action in the greenhouse (low LI) and incomplete dominant to nearly recessive gene action in the nursery (high LI). Incomplete dominance was expressed by heterogeneous to susceptible reaction types of heterozygote plants (S_H4s_H4), under high LI.Some ecological and breeding aspects of the observed effect of LI on resistance to coffee leaf rust are discussed.Samenvatting Resistentie van koffie tegen fysio II vanHemileia vastatrix werd getoetst in milieus bij lichtintensiteiten (LI), die varieerden van 17 tot 100% van de totale instraling. Negen behandelingen, bestaande uit de combinaties van drie niveaus van LI vóór inoculatie en drie ná inoculatie, werden toegepast op zaailingen van het vatbareCoffea arabica ras Mundo Novo. Toenemende LI vóór inoculatie veroorzaakte een significante toename in lesiedichtheid, terwijl het tegenovergestelde werd waargenomen bij de behandeling na inoculatie. Maximale verschillen in lesiedichtheid waren drievoudig. De interactie tussen behandelingen vóór en ná inoculatie was ook significant. Bij extreem hoge LI ná inocultie trad necrose van de lesies op.Genotypen van de Icatu populatie en van hetC. canephora ras Kouillou, met verschillende ziektescores in het veld, werden beproefd in verschillende milieus, waarbij een constante LI voor en na inoculatie werd toegepast. De resistentie van de meeste genotypen kwam beter tot uiting bij lage LI dan bij hoge LI, wat ook waargenomen werd voor het controle ras Mundo Novo. Bij het ras Kouillou werden de dichtheid van sporulerende lesies, de latentieperiode en het reactietype significant beïnvloed door LI en genotype. De interactie tussen LI en genotype was ook significant voor dichtheid van sporulerende lesies en voor reactietype, voornamelijk doordat het meest resistente genotype niet, of in de omgekeerde richting, beïnvloed werd door LI.De expressive van het resistentiegen S_h4 bleek ook afhankelijk van het milieu. Waarnemingen aan een uitsplitsende F2-populatie duidden op een dominante genwerking in de kas (lage LI) en een incompleet dominante, of bijna recessieve, genewerking in de kwekerij (hoge LI). Deze incomplete dominantie uitte zich d.m.v. heterogene tot vatbare reactietypes van heterozygote planten (S_H4s_H4) onder hoge LI.Enkele ecologische en veredelingstechnische aspecten van de waargenomen invloed van LI worden besproken.     

Temperature requirements for germination,germ tube growth and appressorium formation of urediospores of Hemileia vastatrix

Requirements for germination, germ tube growth and appressorium formation of urediospores ofHemileia vastatrix, the causal organism of coffee leaf rust, were investigated by applying treatments with constant temperatures, ranging from 10 to 31°C, and with variable temperatures. Observations were made after 4, 6, 8 and 24 h of incubation of the spores in distilled water on glass slides. The lower and upper limits for germination were estimated to be 13 and about 30°C, respectively. Germination was quickest at 22 to 28°C, whereas appressoria formed more rapidly at 13 to 16°C. After 24 h of incubation, a broad optimum, from 16 to 28°C, was observed for germinationn and total appressorium formation. The shape of the appressoria was torpedo-like or roundish at 13 to 19°C, whereas at higher temperatures their shape was predominantly irregular. Germ tube length increased linearly with time, with an optimum at 19 to 22°C. The degree of branching of the germ tube was positively correlated with germ tube length. Germination percentage and appressorium formation were higher on leaf disks than on glass slides. With variable temperatures, significant correlation were found between temperature sum and percentage of germination, appressorium formation and germ tube length.The results show that germination and appressorium formation can occur at lower temperatures than reported in literature. Penetration byH. vastatrix seems te be realized most rapidly in nature, if, after wetting of the spores, a decline in temperature occurs from initially ca. 23 to 17°C within a few hours. Such conditions prevail during evening and early night in many areas where Arabica coffee is grown.Samenvatting De temperatuurbehoeften voor kieming en appressoriumvorming van uredosporen vanHemileia vastatrix werden onderzocht door incubatie bij constante temperaturen, 10 tot 31°C, en variabele temperaturen. Waarnemingen werden gedaan na 4, 6, 8 en 24 uur incubatie van de sporen in gedestilleerd water op glasplaatjes. De onder- en bovengrens voor kieming bleek bij 13 en ca 30°C te liggen. De kieming kwam het snelst op gang bij 22 tot 28°C en de appressoriumvorming bij 13 tot 16°C. Na 24 uur incubatie werd een breed optimum waargenomen van 16 tot 28°C voor het kiempercentage en het totale aantal appressoria. De vorm van de appressoria was torpedoachtig of rond bij 13 tot 19°C, terwijl bij hogere temperaturen de vorm meestal onregelmatig was. De lengte van de kiembuis nam vrijwel rechtlijnig toe met de tijd, met een optimum van 19 tot 22°C. De vertakkingsgraad van de kiembuizen was positief gecorreleerd met de kiembuislengte.Kieming en appressoriumvorming verliepen beter op de bladschijven dan op glasplaatjes. Bij de behandelingen met variabele temperaturen werden significante correlaties waargenomen tussen de temperatuursom en de kiembuislengte en ook tussen de temperatuursom en het kiem- en appressoriumpercentage.De resultaten tonen aan dat kieming en vooral appressoriumvorming bij lagere temperaturen kan plaatsvinden dan verondersteld in de literatuur. Ze suggereren dat penetratie vanH. vastatrix het snelst gerealiseerd kan worden in de natuur als, na bevochtiging van de sporen, een temperatuurdaling plaatsvindt van aanvankeljk ca. 23 naar ca. 17°C binnen een paar uur. Deze conditie komt veelvuldig voor in de namiddag en het begin van de nacht in vele gebieden waar Arabica-koffie wordt verbouwd.     

马铃薯内生细菌的分离及环腐病拮抗菌的筛选鉴定

 本研究从大同、太原和内蒙古等地采集马铃薯块茎,分离到240株内生细菌,通过离体抑菌作用测定,共得到55株对环腐病菌有拮抗作用的菌株,占菌株总数的22.9%,抑菌圈半径最大的可达13 mm。按抑菌圈半径大小将拮抗菌分为强、中、弱三类。从中筛选出9个对环腐病等病菌具有较强拮抗作用的内生菌株进行了细菌学鉴定,结果表明:118为荧光假单胞生物型V (Pseudomonas fluorescens biovar V);110为短小芽孢杆菌(Bacillus pumilus);085为嗜热脂肪芽孢杆菌(Bacillus stearothermophilus);069为草生欧文氏菌(Erwinia herbicola);043为草莓黄单胞菌(Xanthomonas fragariae);A-10'、T3为芽孢杆菌属(Bacillus);H1-6为荧光假单胞菌(Pseudomonas fluorescens);116为短小杆菌属(Curtobacterium)。     

The use of leaf disk inoculations in assessing resistance to coffee leaf rust (Hemileia vastatrix)

The suitability of inoculations of leaf disks of 1.8 cm diameter in determining resistance of coffee toHemileia vastatrix, the causal agent of coffee leaf rust, was studied. Results obtained by this method were similar to those obtained by greenhouse tests with respect to reaction types of coffee plants with complete and/or major gene resistance. The efficacy of the method in assessing incomplete resistance was tested on 19 plants ofCoffea canephora cv. Kouillou, which varied in level of disease in the field. Four series of inoculation were carried out in four different months of the year, and six components of resistance were assessed. The analysis of multiple correlation, applied to the average data of the four series, indicated that 79% of the variation in disease in the field could be explained by the observations in the leaf disk test. For the individual series this percentage varied from 58 to 70. The coefficients of correlation between the six components were significant and high. The percentage of leaf disks with sporulating lesions was found to be the most suitable component for assessing incomplete resistance.The number of lesions per leaf disk was affected substantially by the hour of the day of leaf collection and by light intensity in the field. No effect was observed of the size of the disks (1 to 2 cm in diameter) and of the leaf wetness period after inoculation (24 and 48h). Results were more consistent when the inoculum was applied in droplets of 0.025 ml than when the inoculum was sprayed onto the disks. No genotype x treatment interactions were observed for the hour of leaf collection, for the size of the leaf disk, for the inoculation method or for the leaf wetness period.It is concluded that the leaf disk method, if adequately standardized, can be a very useful tool in breeding for coffee leaf rust resistance and also in basic research on the coffee —H. vastatrix relationship.Samenvatting De geschiktheid van inoculaties van bladschijven van 1.8 cm diameter voor de bepaling van resistantie van koffie tegenHemileia vastatrix, de veroorzaker van koffieroest, werd nagegaan. Voor het bepalen van het reactietype van koffieplanten, met complete en/of monogene resistentie, bleek de bladschijfmethode resultaten op te leveren die vergelijkbaar waren met die van kasproeven. De geschiktheid van de methode voor het bepalen van onvolledige resistentie werd beproefd bij 19 planten, behorende tot het ras Kouillou vanCoffea canephora, die varieerden in veldaantasting. Vier inoculatie series werden uitgevoerd in vier verschillende maanden van het jaar, waarbij zes resistentie-componenten werden bepaald. Multipele correlatie voor de gemiddelden van de vier series toonde aan dat 79% van de variatie in veldaantasting te verklaren was door de waargenomen componenten in de bladschijftoets. Voor de individuele series varieerde dit percentage tussen de 58 en 70. De zes componenten vertoonden onderling een sterke mate van correlatie. Het percentage bladschijven met sporulatie bleek de meest geschikte component te zijn voor het schatten van onvolledige resistentie.Het aantal lesies per bladschijf werd duidelijk beïnvloed door het uur van de dag waarop de bladeren werden geplukt en de hoeveelheid licht waaraan de bladeren waren blootgesteld in het veld. De grootte van de bladschijven (1 à 2 cm in diameter) en de bladnatperiode na inoculatie (24 en 48 uur) bleken hierop geen effect te hebben. De inoculatiemethode, waarbij druppels van 0,025 ml werden gebruikt, bleek meer consistente resultaten te geven dan de methode waarbij het inoculum op de bladschijven werd gespoten. Er werd geen genotype x behandeling interactie waargenomen voor het uur van de dag waarop de bladeren werden geplukt, de grootte van de bladschijven, in inoculatiemethode en de duur van de bladnatperiode.Geconcludeerd wordt dat de bladschijfmethode, mits toegepast in gestandaardiseerde vorm, een zeer bruikbaar hulpmiddel kan zijn bij de veredeling van koffie op roestresistantie en bij het onderzoek naar de relatie tussen koffie enH. vastatrix.     

月季根癌病病原菌分离及抗病资源初步筛选

从月季品种金玛丽、曼海姆、杏花村、梅郎口红的根部肿瘤组织中纯化了9株分离物,根据其在MW选择性培养基上的单菌落形态初步判断其为根癌土壤杆菌。以根癌土壤杆菌高度保守的virD2和ipt基因的部分序列设计引物对分离的菌株进行PCR检测,其中有6株菌株能扩增出virD2和ipt基因片段,为根癌土壤杆菌毒性菌株。采用针刺涂抹法接种向日葵、荷花蔷薇幼茎,6株菌株均能在供试植株上形成肿瘤,一个月后肿瘤大小有显著差异,说明分离获得的根癌土壤杆菌株毒性有差异。以强毒性菌株J-5-1在田间接种了部分蔷薇属野生资源,以鉴定其对根癌病的抗性。根据发病率、肿瘤大小、肿瘤干质量以及感病后植株生长状况将植物的根癌病抗性分为高度抗病、中度抗病、中度感病、高度感病4个类型。     

Hypersensitive cell death and post-haustorial defence responses arrest the orange rust (Hemileia vastatrix) growth in resistant coffee leaves

The growth of a coffee orange rust fungus (Hemileia vastatrix Berk and Br.) isolate (race II) and the sequence of responses it induced in leaves of resistant Coffea arabica L. and C. congensis Froehner as well as on a susceptible C. arabica were investigated cytologically and biochemically. The percentages of germinated urediospores and of appressoria formed over stomata as well as the fungal growth inside leaf tissues were similar in resistant and susceptible leaves until the 3rd day after the inoculation. In the susceptible leaves, at the majority of the infection sites (70%) the fungus pursued its growth without apparent inhibition while in the resistant leaves the fungus ceased its growth with higher frequency (34% in C. arabica and 54% in C. congensis) after the formation of at least one haustorium. The first signs of incompatibility, detected 2 days after the inoculation, were cytologically expressed by hypersensitive host cell death (HR), host cell wall autofluorescence and haustoria encasement with callose and β-1,4-glucans. Biochemically, two peaks of phenylalanine ammonia-lyase (PAL) activity were detected by 2 and 5 days after the inoculation. The 1st peak coincided with the early accumulation of phenolic compounds and with the beginning of cell death. The 2nd peak could be related to later accumulation of phenols and the lignification of the host cell walls. About 5–7 days after the inoculation, ultrastructural observations revealed the accumulation of a material partially crystallized in the intercellular spaces around the senescent hyphae, next to dead host cells and in close association with the middle lamella that initially labelled for pectins. It also contained polysaccharides and phenolic-like compounds. Cellulose, hemicellulose, extensins, hydroxyproline-rich glycoproteins and proteins were not detected. The hypertrophy of the host cells in the infection area were also observed around 12 days after the inoculation corresponding macroscopically to the reaction flt.In susceptible plants, cell death was also observed 3 days after the inoculation but only in a reduced percentage of infection sites in which the fungus aborted at an early stage. A late haustorium encasement and stimulation of PAL activity were also observed but these delayed host responses did not prevent fungal growth and sporulation.The intercellular material, only observed in the resistant plants, is here reported for the first time and although its role is unknown it might be the result of plant cell death.     

Qualitative and quantitative variation in pathogenicity of races of coffee leaf rust (Hemileia vastatrix) detected in the State of São Paulo,Brazil

Between 1977 and 1981, seven qualitatively distinct new races of coffee leaf rust (Hemileia vastatrix) were detected in breeding plots in the State of São Paulo, Brazil. Four races carry complex virulence against the resistance genes S_H1, S_H2, S_H4 and S_H5 ofCoffea arabica. Three races match unidentified resistance genes ofC. canephora. Two of these were isolated from cv. Kouillou and one from the hybrid population Icatu. Pending further identification, these races were indicated by the number of their type cultures (Is. 2, 10 and 11). Is. 2 and Is. 10 showed extra virulence to some coffee genotypes and decreased virulence to other coffee genotypes, suggesting stabilizing selection. Three rust isolates were detected which differed quantitatively from the common rust race II. Is. 1 was moderately virulent to the coffee differential for S_H3 in the laboratory but avirulent in the greenhouse, indicating a host x pathogen x environment interaction. Is. 3 and 12 showed levels of virulence intermediate between race II and Is. 2 and 10, respectively. The results show that incomplete resistance of coffee toH. vastatrix, at various levels, can be race-specific. The nature of race formation of coffee leaf rust in Brazil and breeding strategies for obtaining durable resistance are discussed.     

Evidence for hyperparasitism of coffee rust (Hemileia vastatrix) by the entomogenous fungus, Lecanicillium lecanii, through a complex ecological web

The entomogenous fungus, Lecanicillium lecanii is hyperparasitic on Hemileia vastatrix , the cause of coffee leaf rust in the laboratory, and has frequently been observed attacking it in the field. The existence of a complex ecological web involving the spatially clustered mutualism of an ant ( Azteca instabilis ) with a scale insect ( Coccus viridis ), where the scale insect was infected by L. lecanii , prompted a search for a spatial correlation between the attack of L. lecanii on the scale insect and the incidence of rust in a commercial coffee crop. A weak but statistically significant effect of hyperparasitic control of coffee rust was observed on two distinct scales: in a 45-ha plot and on a scale of approximately 10 m. It was concluded that this effect was linked to an indirect effect of the ant–coccid mutualism, where L. lecanii was a parasite of the coccid.     

Use of fungal antagonists for biocontrol of damping-off and sclerotinia diseases

Two mycoparasites, Pythium oligandrum and Coniothyrium minitans, have been tested for their ability to act as disease biocontrol agents. P. oligandrum oospores, grown in a cane molasses liquid medium and coated onto cress and sugar-beet seeds using commercial thin-film or pelleting techniques, gave significant control of damping-off in cress and sugar-beet caused by Pythium ultimum and Aphanomyces cochlioides respectively, in glasshouse pot trials. In some cases, the control was equivalent to fungicide drenches or standard fungicide seed treatments, but little control was achieved with any treatment when the pathogen inoculum potential in the soil was high. Pre-planting application of a solid substrate preparation of C. minitans gave reproducible control of sclerotinia disease in the glasshouse. The degree of control was equivalent to that achieved with regular foliar sprays of vinclozolin, when there was less than 40% disease in the control plots naturally infested with Sclerotinia sclerotiorum. At higher disease levels biocontrol was lost. Nevertheless, C. minitans survived in the soil for over one year and continued to degrade sclerotia and reduce apothecial production. The commercial potential of these biocontrol agents is discussed.     

Biological control of Rhizoctonia solani on potatoes by antagonists. 4. Inoculation of seed tubers with Verticillium biguttatum and other antagonists in field experiments

Inoculation of seed potatoes with the mycoparasiteVerticillium biguttatum, isolate M73 (combined withGliocladium roseum in 1981, either alone or mixed with isolate M180 plus antibiotics-producing isolates ofAzotobacter chroococcum in 1982) repeatedly proved successful in reducingRhizoctonia solani on stolons and stems. In field experiments, this ultimately led to a reduced formation of sclerotia on new tubers, particularly in neutral sandy loam and clay loam soils. In 1981 inoculation with antagonists led, when compared with no inoculation, to average reductions of 22 and 42% for the harvest from clean, and 15 and 26% for the harvest from infected seed tubers grown on slightly acid sandy soils and on neutral loam soils, respectively. The harvest from clean, inoculated seed tubers had the lowest sclerotium index. In 1982 inoculation of seed tubers planted in slightly acid sandy soils gave reductions of the sclerotium index of up to 22%. In the neutral marine loam soils considerable reductions were often achieved, viz., in slightly infected loams 51–68% and in rather heavily infected ones 4–43%. Chemical disinfection of seed tubers proved effective only in loam soils that were slightly infested withR. solani. In both years inoculation of seed tubers with antagonists led to significantly lower sclerotium indices of the harvest (p=0.1% in 1981; p=5% in 1982). V. biguttatum was present more frequently and in greater densities on stems and stolons of plants from inoculated than from non-inoculated seed tubers. The latter were colonized by wildV. biguttatum strains from the soil, apparently less effective antagonists.Early in the season, the soil temperature was too low for growth ofV. biguttatum. Nevertheless, inoculation of tubers that were planted early resulted in a considerable cotrol ofR. solani.Samenvatting Het beënten van poters met de opRhizoctonia solani parasiterende schimmelVerticillium biguttatum isolaat M73 in combinatie metGliocladium roseum (1981) of metV. biguttatum M73 alleen of in combinatie met isolaat M180 plus antibiotische isolaten van de bacterieAzotobacter chroococcum (1982), bleek effectief in het terugdringen of het onderdrukken vanR. solani op stengels en stolonen en het verminderen van de aantasting.Beënting van het pootgoed leidde tot een vermindering van de sclerotium (lakschurft)-vorming op de nieuwe knollen, vooral in klei-en zavelgronden. In 1981 leidde beënting van poters tot reductie in de sclerotiumvorming van gemiddeld 22 en 42% voor de oogst uit schoon en 15 en 26% voor de oogst uit besmet pootgoed geteeld op respectievelijk zandgrond en klei- en zavelgrond.In 1982 leidde beënten van de poters uitgeplant in licht zure zandgrond tot een gemiddelde reductie van de sclerotiumindex van de oogst van 22%. In zwaar besmette zandgrond trad evenwel geen reductie op; de infectiedruk was hier te groot. In de neutrale zavel- en kleigronden, vaak ook in de zwaarder besmette percelen werden aanzienlijke reducties bereikt, in de licht besmette gemiddeld 51–68% en in de zwaarder besmette 4–43%. Ontsmetten van pootgoed bleek alleen effectief in percelen die licht metR. solani waren besmet.In beide jaren bleek beënten van pootgoed met antagonisten te resulteren in een significant lagere sclerotiumindex van de oogst (p=0,1% in 1981; p=5% in 1982). V. biguttatum was veel vaker en meer aanwezig op de ondergrondse stengeldelen en stolonen van planten uit beënt pootgoed dan op die van niet beënte poters. De laatsten werden gekoloniseerd door wilde stammen vanV. biguttatum uit de grond, die vaak minder effectieve antagonisten waren. Beënting van vroeg gepote knollen — als de temperatuur nog te laag is voor de groei vanV. biguttatum — leverde toch gunstige resultaten op.     

Combination and rotation of bacterial antagonists to control powdery mildew on pumpkin

Journal of Plant Diseases and Protection - Mixtures of micro-organisms may lead to more effective control of plant pathogens than biocontrol with single species, but little data exists on the...     

Selection of antagonists from compost to control soil-borne pathogens

Journal of Plant Diseases and Protection - One-hundred and one microorganisms have been selected and tested for their antagonistic activity towards soil-borne plant pathogens from a compost...     

Recombination and selection in populations of plant pathogens

A theoretical model has been used to study the dynamics of the frequencies of the following: a virulence gene which is selected by part of the host plant population; an unnecessary virulence gene, which is not required for infection of the host; and the gametic disequilibrium between the two genes. If the two genes are not initially in gametic equilibrium, the frequency of the unnecessary virulence may be altered greatly by hitch-hiking selection, because of the increased frequency of the selected virulence. The hitchhiking effect is strongest if reproduction is entirely asexual, but can still be significant if the frequency of recombination is less than the fraction of the host population which consists of selectively resistant plants. The frequency of recombination may be reduced if reproduction is partly clonal, rather than fully sexual, or if the two genes are linked. Selection against unnecessary virulence may give rise to complex dynamics of both virulence alleles; in particular, the frequency of an unnecessary virulence can rise substantially, by hitch-hiking selection, even if there is some sex or recombination. The direction in which the unnecessary virulence's frequency changes depends on the sign of the gametic disequilibrium between it and the selected gene, and on the existence of selection against unnecessary virulence. If there is no such selection, the long-term dynamics of genotype frequencies in a largely asexual pathogen population may be unpredictable. Consequently, disease control strategies based on planned replacements of one resistance gene by another are unlikely to be effective.