Production of conidia by Peronosclerospora sorghi on sorghum crops in Zimbabwe

Factors affecting the production of conidia of Peronosclerospora sorghi , causing sorghum downy mildew (SDM), were investigated during 1993 and 1994 in Zimbabwe. In the field conidia were detected on nights when the minimum temperature was in the range 10–19°C. On 73% of nights when conidia were detected rain had fallen within the previous 72 h and on 64% of nights wind speed was < 2.0 m s⁻¹. The time period over which conidia were detected was 2–9 h. Using incubated leaf material, conidia were produced in the temperature range 10–26°C. Local lesions and systemically infected leaf material produced 2.4–5.7 × 10³ conidia per cm². Under controlled conditions conidia were released from conidiophores for 2.5 h after maturation and were shown to be well adapted to wind dispersal, having a settling velocity of 1.5 × 10⁻⁴ m s⁻¹. Conditions that are suitable for conidia production occur in Zimbabwe and other semi-arid regions of southern Africa during the cropping season.     

Pollen mimicry during infection of blueberry flowers by conidia of Monilinia vaccinii-corymbosi

Infection of blueberry flowers by Monilinia vaccinii-corymbosi, which causes mummy berry disease, occurs via the gynoecium and involves conidial germination on the stigmatic surface followed by hyphal ingress into the stylar canal and subsequent colonization of the ovary. The extent to which these events mimic pollen–pistil interactions during pollination was investigated. Similar to blueberry pollen tubes, conidial germ tubes of M. vaccinii-corymbosi adhered selectively to imprints of stylar transmitting tract tissue on nitrocellulose membrane, with adhesion in both cases occurring at the tips of the tubes. By contrast, hyphae of the related Monilinia fructicola, which is nonpathogenic on blueberry and does not cause gynoecial infection, adhered indiscriminately to the entire membrane. Using monoclonal antibodies, the presence of epitopes of esterified and unesterified pectins and of arabinogalactan proteins (AGPs), which have been implicated in adhesion and pollen tube guidance in other plant species, was documented on blueberry pollen tubes in vitro. Epitopes of certain AGPs, but not of pectins, were also localized on conidia and hyphae of M. vaccinii-corymbosi. However, such epitopes were also detected on M. fructicola, suggesting that they are unlikely to be a discriminatory factor between fungi capable or not capable of gynoecial infection. Microscopic observation of inoculated pistils showed that similar to pollen tubes, hyphae of M. vaccinii-corymbosi tracked the lobes of the stylar lumen, grew directionally (i.e. with very limited branching) in close proximity to cells of the stylar adaxial epidermis and to one another, and were surrounded by extracellular matrix. By contrast, hyphae of M. fructicola, while being able to ingress into the style, branched profusely within the stylar canal, showing no directional growth or affinity to specific regions of the lumen. We propose these results as evidence of specialized opportunism by M. vaccinii-corymbosi, whereby fungal hyphae appear to mimic host pollen tubes and take advantage of an infrastructure intended to support host reproduction in order to facilitate infection of the ovary.     

Patterns of release of the secondary conidia of Claviceps africana, the sorghum ergot pathogen in Australia

Trials were conducted in southern Queensland, Australia between March and May 2003, 2004 and 2005 to study patterns of hourly and daily release of the secondary conidia of Claviceps africana and their relationships with weather parameters. Conidia were trapped for at least one hour on most (> 90%) days in 2003 and 2004, but only on 55% of days in 2005. Both the highest daily concentration of conidia, and the highest number of hours per day when conidia were trapped, were recorded 1–3 days after rainfall events. Although the pattern of conidial release was different every day, the highest hourly conidial concentrations occurred between 10·00 hours and 17·00 hours on 73% of all days in the three trials. Hours when conidia were trapped were characterized by higher median values of temperature, windspeed and vapour pressure deficit, lower relative humidity, and leaf wetness values of 0%, than hours when no conidia were recorded. The results indicate that fungicides need to be applied to the highly ergot-susceptible male sterile (A-) lines of sorghum in hybrid seed production blocks and breeders' nurseries as soon as possible after rainfall events to minimize ergot severity.     

Effect of inoculum composition on infection of French bean leaves by conidia of Botrytis cinerea

Inoculation of leaves of French bean (Phaseolus vulgaris) with sprays or small drops of a suspension of conidia ofBotrytis cinerea gave rise to spreading lesions, lesions remaining restricted in size or to no visible necrosis. The type of reaction depended on the composition of the inoculum. In studies with drop inoculations with buffered inocula some of the factors involved were analyzed. The formation of spreading lesions depended on pH, type and molarity of the buffer, presence of glucose, and concentration of conidia in the inoculum. If the phosphate buffer used in most of the inocula was replaced by monobasic phosphate, similar results were obtained. The reactions were not influenced by the proportion of K⁺ or Na⁺ ions in the phosphate buffer. Inoculations with conidia suspended in a solution of 0.067 M phosphate buffer (pH 5.0) or monobasic phosphate and 0.11 M glucose always evoked a susceptible reaction, i. e. the formation of spreading lesions.     

番茄斑萎病毒系统侵染我国大蒜

番茄斑萎病毒Tomato spotted wilt tospovirus(TSWV)是严重危害世界经济作物的一种病毒,寄主范围广泛。我们在研究中发现番茄斑萎病毒能侵染我国大蒜Allium sativum L.。利用摩擦接种法将TSWV接种到健康大蒜上,结果显示:接种14 d后大蒜新生叶片出现褪绿和白色斑点症状。ELISA检测显示大蒜叶片汁液与TSWV的单克隆抗体产生血清学反应,采用TSWV N基因的引物对大蒜叶片总RNA进行RT-PCR,结果扩增出约800 bp的条带,在NCBI上BLAST显示与TSWV YN5576的同源性最高,为98.52%。这些数据表明番茄斑萎病毒系统侵染我国大蒜。     

Epidemiology of brown rot (Monilinia fructigena) on apple: infection of fruits by conidia

Effects of fruit maturity, wound age, temperature and the duration of wetness periods on infection of apple fruits by conidia of the brown rot fungus, Monilinia fructigena , were studied. Inoculation of fruits on potted apple trees and harvested mature fruits showed that wounding was essential for infection by M. fructigena . On potted trees, there was a significant difference between the susceptibility of cvs Cox and Gala and this difference depended on wound age. The incidence of brown rot was affected greatly by fruit maturity and wound age. Wounds on younger fruits were more resistant to infection than those on older fruits, whilst the older the wound, the more resistant it was to infection. Furthermore, the degree of wound age-related resistance was greater on younger fruits than on older fruits. These relationships were well described by regression models. The effect of the duration of wetness periods was very small: increasing the duration of wetness periods reduced the incidence of brown rot on older wounds. For detached fruits, all those wounded were rotted after inoculation, except for those in two treatments under 20°C on fruits with wounds which were 8 days old. The incubation period of the fungus was generally very short. Wound age was the single most important factor influencing the length of the incubation period; the incubation period increased as wound age increased.     

Effects of temperature and continuous and interrupted wetness on the infection of pear leaves by conidia of Venturia nashicola

Experiments were conducted to determine: (i) the effects of temperature and duration of continuous wet periods on the infection of pear seedlings by conidia of Venturia nashicola , the causal agent of pear scab; and (ii) the effects of the length and temperature of dry interrupting periods on the mortality of infecting conidia. Average number of scab lesions per leaf increased with increasing duration of wetness. Logistic models adequately described the change in the average number of scab lesions per leaf at 5, 10, 15, 20 and 25°C over the wetness duration. At 30°C, only a few lesions developed. Simple polynomial models satisfactorily described the relationship of the three logistic model parameters (maximum number of lesions, rate of appearance and the time to 50% of the maximum number of lesions) with temperature. The optimum temperature for infection was found to be approximately 20°C. The relationship between mortality and the length of a dry period interrupting an infection process can be satisfactorily described by an exponential model. The rate of mortality at 10, 16 and 22°C did not differ significantly, but was significantly less than that at 28°C.     

Accumulation of sorghum phytoalexins induced by Colletotrichum graminicola at the infection site

Microspectrophotometry was performed on intact, pigmented vesicle-like inclusions within living sorghum cells that were accumulating phytoalexins as a response to attempted fungal infection. The results indicate that the deoxyanthocyanidin phytoalexins are present in inclusions. Moreover, the phytoalexin concentration within a single inclusion, based on luteolinidin, was calculated to be 0·15 m. The amounts of luteolinidin and apigeninidin in cells involved in the phytoalexin response at individual infection sites were also determined. The data showed that luteolinidin accumulated to levels of 0·48–1·20 ng/cell whereas apigeninidin accumulated to levels of 0·24–0·91 ng/cell. The results of both analyses confirmed that at the infection site the deoxyanthocyanidins accumulate to levels in substantial excess of those required for inhibition of the fungus Colletotrichum graminicola.     

Systemic infection of petunia by mechanical inoculation with tomato golden mosaic virus

Samenvatting Aangetoond werd datPetunia hybrida systemisch kan worden geïnfecteerd met het tomato golden mosaic virus (TGMV), een virus dat behoort tot de groep van de geminivirussen. Mechanische inoculatie van petuniaplanten met TGMV gaf in de systemisch geïnfecteerde bladeren symptomen, die eerder in een aantal andere Solanaceae waren waargenomen. Daar in eerdere proeven petunia niet met TGMV kon worden geïnfecteerd en DNA-replicatie en symptoomontwikkeling wel optrad in, voor de beide genomen van het virus, transgene planten, werd gesuggereerd dat het hier een geval betrof van uitbreiding van de waardplantenreeks.De hier gepresenteerde resultaten kunnen echter tot andere conclusies leiden. Het is namelijk mogelijk, dat bepaalde F₁-hybriden van petunia resistenter zijn tegen het virus. Verschillen in de symptoomontwikkeling zijn echter ook niet uit te sluiten en zouden veroorzaakt kunnen worden door premunitie als gevolg van de aanwezigheid van het manteleiwit in opnieuw geïnfecteerde cellen.     

The role of infection time in the differential response of sorghum cultivars to Striga hermonthica infection

Striga hermonthica is an important parasitic weed that severely reduces yields of sorghum in sub‐Saharan Africa. Pot experiments with the sensitive sorghum cultivar CK60‐B and the tolerant Tiémarifing were conducted in 1999 and 2000 to investigate the role of infection time on the interaction between sorghum and Striga hermonthica. Timing of Striga inoculation was used to establish delays of one and two weeks in first attachment of the parasite. In 1999, early Striga inoculation resulted in a relatively early first Striga attachment on CK60‐B. Although first infection of Tiémarifing occurred one week later, an identical final number of emerged Striga plants was observed. Plants of CK60‐B were more severely affected and supported a higher total Striga biomass. Only with this cultivar the interaction between host and parasite was significantly affected by delayed infection. Parasite biomass was most sensitive and already significantly reduced following a 1‐week delay in infection time. With a further 1‐week delay, an additional reduction in parasite biomass was accompanied by a strong and significant increase in total and panicle dry weight of the host plant. In 2000, first infection of CK60‐B was relatively late and occurred simultaneously with first infection of Tiémarifing and no significant effect of delayed infection on Striga biomass or host‐plant performance was observed. The results indicate that the influence of delayed infection strongly depends on actual infection time and confirm that earlier observed differences in time of first infection between the two cultivars do contribute to the more tolerant response of Tiémarifing to Striga infection.     

Influence of environmental conditions in a glasshouse on conidia ofBotrytis cinerea and on post-harvest infection of rose flowers

Quantification and horizontal distribution of air-borne inoculum ofBotrytis cinerea in a rose crop in a glasshouse of 300 m² was studied in 1991 and 1992. Conidia ofB. cinerea were caught in spore traps consisting of an agar medium selective forB. cinerea in Petri dishes placed within the crop, at flower height 1 m above the ground. Spore catches were counted as colonies, after incubation. Lesions due to conidial infection were counted on petals of rose flowers, also after incubation. Relative humidity (RH) and temperature within the glasshouse and global radiation and windspeed outside were recorded during the experiments. The horizontal distribution ofB. cinerea in a rose crop grown under glass was fairly uniform in both years. In 1991 a clear seasonal pattern in the number of colonies could not be found. In 1992 the number of colonies were high in August, September and October. The number of lesions on rose flowers showed a distinct pattern in both years. In August, September and October many lesions were counted whereas in the other months few lesions appeared. In linear regression analysis, variation in numbers of colonies (spore catches) could not be explained by environmental factors recorded during the experiments. Linear regression accounted for 76 and 63% of the variation in the number of lesions on rose flowers in 1991 and 1992, in terms of relative humidity (positively correlated), global radiation outside the glasshouse (negatively correlated) and, numbers of colonies on spore traps (positively correlated). The results in the rose crop suggest that RH, global radiation and spore density in glasshouses are important variables in regulating the numbers of lesions during storage and transport. The numbers of spores in glasshouses are dependent on the production system. A glasshouse with a system resulting in wet dead tissue on the ground give higher amount of spores in the glasshouse air and through that high numbers of lesions on flowers. On roses outside the glasshouses very high numbers of lesions were counted sometimes, mostly during and after rain showers, as a result of rain-deposition of spores onto the flowers.     

Measuring cysteine biosynthesis activity from serine in extracts from sorghum, corn and grass weeds, and their metolachlor susceptibility

In vitro assay procedures for measuring the activity of cysteine biosynthesis from serine (CBS), which is a coupled reaction catalyzed by serine acetyltransferase and cysteine synthase, were developed using crude extracts from sorghum shoots. Cysteine biosynthesis from serine activity was dependent on acetyl‐CoA concentrations (up to 1.5 mmol L^?1), serine (at least up to 20 mmol L^?1) and sulfide (up to 0.25 mmol L^?1), respectively, and was proportional to the protein concentration in the reaction mixture below 0.4 mg mL^?1. The reaction rate was 6.6 nmol min^?1 per mg of protein during the first 5 min, but increased to 45.6 nmol min^?1 per mg of protein between 30 and 45 min after reaction initiation. Sorghum had the highest CBS total activity (222.4 nmol min^?1 per g of fresh weight), and large crabgrass had the lowest CBS total activity (4.7 nmol min^?1 per g of fresh weight) when CBS activity in shoots was extracted from sorghum, corn, johnsongrass, barnyardgrass, goosegrass, green foxtail and large crabgrass. Similar results were obtained for CBS specific activity (nmol min^?1 per mg of protein). There was no correlation between total CBS activity and susceptibility to metolachlor; however, when corn was excluded, a correlation of R² = 0.690 was found. Flurazole seed treatment (1.25 g per kg of seed) conferred metolachlor resistance by sorghum, and enhanced total CBS activity and non‐protein thiol content by 27 and 61%, respectively. The increase in thiol content presumably contributed to metolachlor tolerance in sorghum. From these results, the difference in CBS activity partially contributes to the selectivity to metolachlor among certain grass species, and to the safening action of flurazole by increasing thiol content.     

Pre‐infection processes of Botryosphaeriaceae spp.: adhesion of conidia to different substrata

Species of Botryosphaeriaceae are important wound pathogens of grapevines as causal agents of botryosphaeria dieback, but the behaviour of their conidia pre‐infection is unknown and may be important for disease development. Adhesion properties of conidia were investigated for Botryosphaeria dothidea, Neofusicoccum luteum and N. parvum on substrata with different affinities for water. Greatest adhesion on any surface was reached after 5 min for isolates N. luteum MM558, B. dothidea 007 and N. parvum G652 (53·1, 54·0 and 50·6%, respectively) and for N. luteum isolate CC445 after 20 min (61·4%). As conidia adhered well to all artificial substrata, it appeared as if the attachment process was nonspecific. Overall, surface wettability did not play a major role in the adhesion of conidia. Spore surface proteins appeared to play a role in the adhesion process because treatment of conidia of N. luteum MM558 with a protease completely prevented adhesion. Histochemical labelling of conidia and germlings with Coomassie brilliant blue (specific for proteins) was positive for all isolates, with a blue ‘halo’ often seen surrounding conidia or near the germ tube emergence point after incubation times conducive to germination. Alcian blue also stained material surrounding conidia after longer incubation times, which indicated that mucopolysaccharide and protein production may be involved in a second phase of adhesion.     

Effects of temperature, relative humidity and duration of wetness period on germination and infection by conidia of the pear scab pathogen (Venturia nashicola)

Experiments were conducted to determine the effects of temperature, relative humidity (RH) and duration of wetness period on in vitro germination of conidia and infection of detached pear leaves by Venturia nashicola , the causal agent of pear scab. Conidia germinated only in near-saturation humidity (RH > 97%). The final percentage germination (24 h after inoculation) at 100% RH without free water was less than half that in free water. Conidia germinated over the range of temperatures tested (5–30°C); the optimum temperature for germination was ≈21°C. Changes in percentage germination of conidia over time were fitted by logistic models at each individual temperature. Polynomial models satisfactorily described the relationships between two (rate and time to 50% of maximum germination) of the three logistic model parameters and temperature. The minimum length of the wetness period for successful infection of detached pear leaves by conidia was observed at several temperatures. The shortest length of wetness period required for infection was 7 h at 22°C. Two polynomial models fitted well the relationship between the minimum wetness duration required for infection, and temperature.     

Triazine absorption by excised corn root tissue and isolated corn root protoplasts

Absorption of four triazine herbicide analogs [ametryn (2-(ethylamino)-4-(isopropylamino)-6-(methylthio)-s-triazine), atrazine (2-chloro-4-(ethylamino)-6-(isopropylamino)-s-triazine), atratone (2-methoxy-4-(ethylamino)-6-(isopropylamino)-s-triazine), and hydroxyatrazine (2-hydroxy-4-(ethylamino)-6-(isopropylamino)-s-triazine)] was compared using excised corn (Zea mays L.) root segments and isolated corn root protoplasts. The tissue absorbed ametryn, atrazine, and atratone for only 20 min. Ametryn and atrazine permeated tissue to passive equilibrium with the ambient solution in 10 min. Atratone permeated to 65 and 82% of passive equilibrium in 10 and 30 min, respectively. In contrast, hydroxyatrazine concentration in tissue was only 15 and 70% of the ambient concentration at 30 min and 24 hr, respectively. However, hydroxyatrazine permeated frozen/thawed tissue to 90% of passive equilibrium in 10 min. Protoplast absorption of ametryn and atratone was complete in 10 sec; hydroxyatrazine absorption by protoplasts did not reach a plateau until 5 min. Protoplasts absorbed the triazines to greater than passive equilibrium. Three kinetically homogeneous pools were detected for ametryn, atrazine, and atratone, whereas elution of hydroxyatrazine produced four pools. The three pools for atrazine were confounded by metabolism of atrazine to hydroxyatrazine. Pools for the triazines could not be identified as the free space, cytoplasm, and vacuole as proposed previously for mineral ions. Although the plasma membrane impeded diffusion of hydroxyatrazine, all analogs penetrated into the symplast.     

亚洲玉米螟危害蛀孔在春玉米上的分布及其与产量损失的关系

亚洲玉米螟1、2代幼虫在河北张家口春玉米上的危害蛀孔 ,分布在雌穗着生节 (含穗柄 )及其上、下1～3节上的百分比分别为89.9%和85.7%。 1代幼虫蛀孔42.0%分布在雌穗下 1～3节 ,26.8%分布在雌穗上1～3节 ,每株平均有1.8个和2.1个蛀孔。玉米品种逐单 1号和自 330分别减产 19.3%和 23.5%。蛀孔数与产量呈极显著负相关 ,每株增加 1个蛀孔 ,两个品种产量损失率分别为 11.04%和 13.17%。雌穗上部 1个蛀孔的产量损失系数为3.9和 2.9,雌穗下部 1个蛀孔为 16.4和 12.4。 2代幼虫蛀孔 45.7%分布在雌穗以上茎节 ,28.5%分布在雌穗以下茎节 ,每株有 1～6个蛀孔时 ,蛀孔数与产量损失相关系数均未达到 5%的显著标准。     

The ultrastructure of germinating conidia of Penicillium digitatum inhibited by sec-butylamine

Electron micrographs of germinating conidia of P. digitatum showed osmiophilic inclusions scattered in the ground plasm or located within membrane-bound vacuoles where they were associated with various membranes and “myelin-like” structures. Cytochemical studies suggested that the osmiophilic inclusions within the vacuoles contained phospholipids and proteins. These phospholipoprotein inclusions may provide precursors needed for the assembly of membrane-bound organelles formed during germination. In conidia germinated in the presence of fungistatic concentrations of sec-butylamine (SBA) the vacuoles enlarged and the phospholipoprotein inclusions decreased in size and appeared to disintegrate and give rise to membrane-bound vesicles. SBA may deprive the conidium of materials normally utilized for membrane formation by the developing organelles.     

玉米抵御玉蜀黍尾孢菌侵入的生理机制

为深入探讨玉米抗灰斑病的机制,以抗/感玉米灰斑病自交系78599-1、OH43Ht N和掖478、K12为材料,采用比色皿法和RT-PCR法相结合研究了接种玉蜀黍尾孢菌毒素后玉米叶片中防御酶活性、丙二醛(MDA)含量及防御酶合成相关基因SOD、CAT、APX和GR的表达量。结果显示,接种该病菌毒素后,抗、感材料叶片中防御酶活性均升高,多数在接种7 d时达到峰值,且抗病材料防御酶活性峰值高于感病材料;供试材料叶片中MDA含量均降低,且抗病材料低于感病材料;供试材料叶片中SOD、CAT和GR的表达量均上升,其中CAT的表达量在78599-1、掖478和K12接种5d时达到峰值,灰度值分别为228.67、161.33和178.00,与其酶活性变化趋势一致;SOD和GR的变化规律与其酶活性变化不一致;APX的表达量仅在OH43Ht N中上升,接种7 d后达到峰值。表明抗病材料调控防御酶活性的能力强,防御酶基因的表达与其酶活性变化存在关联性。     

玉米抵御鞘腐病菌侵染的生理机制

为深入探讨玉米抗鞘腐病的机制,以高抗品种浚单20、9058及中抗品种郑单958、郑58为材料,测定了接种层出镰孢后苯丙氨酸解氨酶、过氧化物酶、超氧化物歧化酶、多酚氧化酶、过氧化氢酶、木质素含量及抗病相关基因PR-1、PR-2a、LOX、MPI和GAPc的表达量.结果显示:接种后不同抗性材料叶鞘部位的防御酶活性均上升,多数在6h达到峰值,且高抗材料酶活性增幅较大;供试材料的木质素含量均上升,且高抗材料的含量高于中抗材料;PR-1、MPI、GAPc和PR-2a等抗病相关基因的表达呈上升趋势,而LOX的表达量变化趋势不明显.研究表明玉米对鞘腐病的抗性与防御酶活性、木质素含量及其抗病相关基因的表达量呈正相关.