小麦的种和品种对白粉病抗性的初步研究

 在对白粉病具不同抗性的11个小麦的种和品种上进行的按种试验表明,在接种后8小时内,分生孢子达到最高萌发率(约为60%),并形成附着孢。在抗病的和感病的小麦叶片上,分生孢子的萌发率和萌发速度没有差异。接种后12小时,在感病的材料上,发现真菌侵入寄主,但在抗病的材料上,发现最早侵入时间,至少要比在感病材料上约晚4小时。接种后36小时,在高感的材料上,萌发的分生孢子有83%已侵入寄主,其中约有70~80%已形成吸胞。在感病、抗病和高抗材料上的侵入率,分别为70,45,27%。形成或开始形成吸胞的分别为55~61,14~27,4~8%。在抗病材料的表皮细胞内,真菌形成的吸胞较小,并有点畸形。在抗病材料的叶面上,真菌的菌丛稀薄,产孢也很少。     

Environmental Factors Affecting Production, Release, and Field Populations of Conidia of Alternaria alternata, the Cause of Brown Spot of Citrus

ABSTRACT Alternaria brown spot, caused by Alternaria alternata pv. citri, affects many tangerines and their hybrids, causing loss of immature leaves and fruit and reducing the marketability of the remaining fruit. Conidial production of A. alternata was greatest on mature leaves moistened and maintained at near 100% relative humidity (RH) for 24 h, whereas leaves that had been soaked or maintained at moderate RH produced few conidia. Conidial release from filter paper cultures and infected leaves was studied in a computer-controlled environmental chamber. Release of large numbers of conidia was triggered from both substrates by sudden drops in RH or by simulated rainfall events. Vibration induced release of low numbers of conidia, but red/infrared irradiation had no effect. In field studies from 1994 to 1996, air sampling with a 7-day recording volumetric spore trap indicated that conidia were present throughout the year with periodic large peaks. The number of conidia captured was not closely related to rainfall amounts or average wind speed, but was weakly related to the duration of leaf wetness. Likewise, disease severity on trap plants placed in the field weekly during 1995 to 1996 was not closely related to conidial numbers or rainfall amounts, but was weakly related to leaf wetness duration. Sufficient inoculum appears to be available to allow infection to occur throughout the year whenever susceptible host tissue and moisture are available.     

玉米大斑病菌人工接种方法研究

 The efficient and accurate method of inoculating Exserohilum turcicum on maize is most important for studying the corn leaf blight. In this study, four approaches including spore suspension spraying, inoculation with grinding infected maize leaves, inoculation liquid inoculum into the leaf whorl and inoculation with inoculated sorghum seeds were compared with natural induction in fields during different growth stages of maize in the greenhouse and field. The results showed that disease incidence of all varieties tested were up to 100% by inoculating with inoculated sorghum seeds during the booting stage (11 to 12 leaf stages). This method will not only guarantee the enough pathogen inocula but also be easy to use, and could be an effective way for disease resistance identification in field and greenhouse.      

A comparison of inoculation techniques for inducing aflatoxin contamination and<Emphasis Type="Italic">Aspergillus flavus</Emphasis> kernel infection on corn hybrids in the field

Aspergillus flavus inoculation techniques were compared on aflatoxin-resistant and -susceptible corn hybrids for inducing aflatoxin contamination andA. flavus kernel infection. A dry carrier technique was comparable to the standard inoculation techniques (the side-needle and a spray technique) in differentiating between the resistant and the susceptible hybrids in the first year of the study. However, only hybrids inoculated with the side-needle technique had statistically different levels of aflatoxin andA. flavus kernel infection in the second year of the study. In a second study, a modified pinbar technique with inoculations near the tip or base of the ear was compared with the side-needle technique. When developing ears were inoculated near the base with the modified pinbar, adequate levels of aflatoxin were induced both years to distinguish between the resistant and susceptible corn hybrids. The modified pinbar technique has the potential of being a useful tool in evaluating corn germplasm for aflatoxin resistance. http://www.phytoparasitica.org posting May 4, 2007.     

摩洛哥脐血橙植株上一种机械传播性病毒的鉴定

 本文采用叶片接种法和枝条接种法对25个柑桔种类体细胞杂种或品种进行了抗脚腐病鉴定,结果表明:体细胞杂种中枳+酸橙、枳+甜橙为高抗类型;酸橙+兰普来檬、枳+印度酸桔;蚝壳刺+哈姆甜橙、酸橙+澳洲指桔为抗性;酸橙+印度酸桔为中抗。建立了叶片接种法的抗病性分级标准。初步分析了柑桔体细胞杂种的抗性规律。     

Ralstonia solanacearum preferential colonization in the shoot apical meristem explains its pathogenicity pattern in tomato seedlings

Ralstonia solanacearum causes a lethal bacterial wilt disease in many plants by colonizing the vascular tissues of the hosts. Upon inoculation of tomato seedlings through either leaf or root, the wilting symptoms occur first at the apical region and then proceed downward along the shoot. The systemic order of the disease initiation and progression in the host, independent of the site of pathogen inoculation, is yet to be investigated. To understand the disease progression more clearly, we have carried out a systematic study of the pathogen localization by GUS staining of inoculated tomato seedlings, at 24-hour intervals from 0 days post-inoculation (dpi) to 5 dpi. In both inoculation methods, pathogen colonization was observed at 1 dpi at the apical meristem as well as the cotyledon leaves, where the disease initiates. As the disease progressed, colonization by the pathogen towards the lower region of the shoot was observed. Disease consistency and pathogenicity magnitude were observed to be higher using the leaf inoculation method than the root inoculation method. Several R. solanacearum transposon-induced mutants that were reduced in virulence by root inoculation but virulent by leaf inoculation were obtained. Using GUS staining, it was observed that these mutants were unable to localize in the shoot region when inoculated in the root. Our study indicates that the apical meristem and the cotyledon leaves are the first regions to be colonized in inoculated tomato seedlings, which might explain the disease initiation from this region.     

玉米、高粱幼苗丝黑穗病菌的侵染部位研究

 应用幼苗分器官涂抹接种的方法,对玉米两个高感自交系进行了丝黑穗病菌侵染部位的精确测定。三年结果一致表明,侵染以胚芽为主,根为次要。在胚芽上,中胚轴的侵染高于胚芽鞘。在根系中,各种根均能侵染,其中以胚根的感染度稍高,但各种不定根和侧根的数量众多,因此在侵染上仍占相当比重。分生区为各器官的有效侵染点,中胚轴的分生区在紧邻胚芽鞘节下,胚芽鞘和中胚轴的相继反复感染,可以导致很高的发病率。出土前的幼龄植株为主要感染阶段,因而决定根系在总体侵染中不居主要地位。扩大对不同抗性的四个自交系和四个单交种作进一步的测定表明,主要侵染器官及其主次关系在不同品种上基本保持与上述高感自交系相同的趋势。     

苹果炭疽叶枯病病原Glomerella cingulata及其侵染过程

为明确苹果炭疽叶枯病病原菌围小丛壳Glomerella cingulata的侵染致病特征,在分离获得该病原菌的基础上,采用形态学观察、ITS序列分析和致病性测定对其进行了鉴定,并利用光学和扫描电子显微镜对病原菌在嘎啦苹果叶片上的侵染过程进行了研究.结果表明,在陕西咸阳地区分离获得的9株病原菌均为围小丛壳G.cingulata.25 ℃下接种9 h后,分生孢子中间产生隔膜,双胞化,并萌发产生芽管和附着胞;24 h后分生孢子的2个细胞均可萌发并形成芽管及附着胞,部分芽管顶端可产生次级分生孢子;48 h后次级分生孢子萌发形成附着胞;72 h后,附着胞下形成的侵染钉可直接入侵寄主,在表皮细胞内形成初生菌丝和次生菌丝,此时叶片表面已出现褐色斑点.接种7 d后叶片病斑处出现分生孢子盘和子囊壳.表明陕西省近年出现的苹果炭疽叶枯病病原菌为围小丛壳G.cingulata,该病菌在嘎啦叶片上的一些特殊侵染行为可能是导致该病害易在短时间内暴发的重要原因.     

Effects of leaf wetness duration, relative humidity, light and dark on infection and sporulation by Didymella rabiei on chickpea

No infection occurred at less than 95% relative humidity (r.h.) when chickpea plants were dried after inoculation with conidia of Didymella rabiei. Infection was significant when the dry leaves were exposed to 98% r.h. for 48 h. When inoculated plants were subjected to different leaf wetness periods, some infection occurred with 4 h wetness, and disease severity increased with wetness duration according to an exponential asymptote, with a maximum value after about 18 h. Germination of conidia and germ tube penetration increased linearly with increasing wetness periods when recorded 42 h after inoculation. With a 24-h wetness period, germination of conidia was first observed 12 h after inoculation and increased linearly with time up to 52 h (end of the experiment). Dry periods immediately after inoculation, followed by 24-h leaf wetness, reduced disease severity; as the dry period increased the severity decreased. Disease severity increased with increasing periods of darkness after inoculation. The number of pycnidia and the production of conidia on infected leaves increased only slightly with high r.h. (either in the light or in the dark), but large increases occurred over an 8-day period when the leaves were kept wet.     

Comparison of different inoculating methods to evaluate the pathogenicity and virulence of Aspergillus niger on two maize hybrids

A two-year field study was conducted to determine the effects of artificial inoculation techniques on the pathogenicity and virulence of Aspergillus niger kernel infection on two maize hybrids. Test plants included in the study were hybrids resistant and susceptible to Aspergillus flavus to determine if the host resistance mechanisms that limited A. flavus infection would also suppress A. niger infection. Ears were inoculated with the silk-channel, side-needle, and spray techniques 7?days after midsilk (50% of the plants in a plot had silk emerging). Ears were also inoculated with a modified-pinbar technique 21?days after midsilk. Kernel infection in 2008 in inoculated plants ranged from 2% to 11% and from 2% to 45% in the resistant and susceptible hybrids, respectively. In 2009, kernel infection in inoculated plants ranged from 13% to 32% and from 10% to 67% in the resistant and susceptible, respectively. The silk-channel, side-needle, and modified-pinbar techniques produced significantly higher levels of kernel infection in the susceptible hybrid in both years than the spray technique. When hybrids were compared, the silk-channel, side-needle, and modified-pinbar techniques induced significantly higher levels of infections in the susceptible hybrid than in the resistant hybrid in 2008 and 2009. The level of A. niger pathogenicity and virulence increased when conidia were placed inside the husks of developing ears by wounding (modified-pinbar and side-needle techniques) or non-wounding (silk-channel technique) inoculation methods. Although A. niger kernel infection was significantly lower in the A. flavus resistant hybrid compared to the A. flavus susceptible hybrid, A. niger infection levels were much higher than A. flavus infection levels typically observed in both of these hybrids in past studies.     

巴西橡胶树品系对棒孢霉落叶病的抗病性鉴定

采用巴西橡胶树棒孢霉落叶病病原菌的分生孢子接种和其毒素接种分别测定了国内天然橡胶主产区9个主栽品系的抗病性,结果表明过滤灭菌毒素划伤接种橡胶离体小叶片可以产生与分生孢子接种相似的症状,而不划伤接种则不产生病害症状;121℃、20 min湿热灭菌毒素接种无论划伤与否均不产生病害症状;分生孢子和病原菌毒素两种接种方法所测橡胶品系的抗病性之间有很好的一致性,说明毒素接种可以用来代替病原菌接种测定橡胶品系的抗病性。抗病性测定结果显示,供试9个巴西橡胶树品系中,PB86、热研88-13、南华1、PR107、热研7-33-97对病原菌表现出高抗,而RRIM600、南强1-97、热研44-9则表现高感,GT1则表现中感。     

Cacao leaf procyanidins increase locally and systemically in response to infection by Moniliophthora perniciosa basidiospores

Flavan-3-ol monomers and oligomers from dimers to decamers were quantitatively determined by HPLC-FLD in leaves of four Theobroma cacao varieties. Cacao flush leaves had higher procyanidin content than mature leaves. Moniliophthora perniciosa basidiospore infection results in an increase in cacao leaf procyanidins in the inoculated leaves, as well as in the uninoculated, adjacent, older leaves on an actively growing branch, resulting in both a local and systemic increase in procyanidin content in response to infection. Procyanidin oligomers increased significantly in leaves of the less susceptible genotype Scavina 6 upon infection, and total procyanidin levels were higher than in Cacao Comum a more disease susceptible genotype. Selection of genotypes with high constitutive and/or inducible content of procyanidins in young flush leaves may be a strategy to develop witches’ broom disease resistant cultivars.     

Effect of method of inoculation, inoculum density and seedling age at inoculation on the expression of resistance of cocoa (Theobroma cacao L.) to Verticillium dahliae Kleb.

Soil drench and stem puncture inoculation were compared as methods for selecting cocoa cultivars with resistance to Verticillium dahliae. Disease progress was more rapid and induced symptoms were more severe following stem puncture and, under glasshouse conditions, differences between cultivars were detected 15 days after inoculation. Moreover, using stem puncture, inoculum densities of 10⁴ conidia/ml were sufficient to differentiate resistant and susceptible cultivars, whereas with the soil drench method, inoculum densities of 10⁷ conidia/ml were necessary. Although a substantially higher proportion of plants were affected by stem puncture inoculation, the resistance of cultivar Pound-7 remained effective at high inoculum densities of 10⁸ conidia/ml. With either method, older seedlings were more susceptible to V. dahliae than younger ones. However, with stem puncture, 15-day-old seedlings were sufficiently susceptible for a valid disease assessment. In contrast, with soil inoculation, 60-day-old plants were required. In a nursery trial with 15-day-old seedlings, seven cocoa genotypes previously selected as resistant, moderately resistant or susceptible to Verticillium dahliae , on the basis of root inoculation, were ranked in the same order when stem punctured. Stem puncture inoculation of young seedlings is cost-effective in terms of time and space, and is therefore recommended for screening of cocoa for wilt resistance, especially in large-scale breeding programmes.     

Influence of Temperature and Wetness Duration on Conidia and Appressoria of Colletotrichum acutatum on Symptomless Strawberry Leaves

ABSTRACT Strawberry leaves (cv. Tristar) inoculated with Colletotrichum acuta-tum conidia were incubated at 10, 15, 20, 25, 30, and 35 degrees C under continuous wetness, and at 25 degrees C under six intermittent wetness regimes. The number of conidia and appressoria was quantified on excised leaf disks. In order to assess pathogen survival, inoculated leaves were frozen and incubated to induce acervular development. Germination, secondary3 conidiation, and appressorial development were significantly (P /= 0.95) related to appressorial populations prior to this treatment and was greatest following periods of continuous wetness. Production of secondary conidia and appressoria of C. acutatum on symptomless strawberry leaves under a range of environmental conditions suggests that these processes also occur under field conditions and contribute to inoculum availability during the growing season.     

Effects of temperature and wetness duration on conidial infection, latent period and asexual sporulation of Pyrenopeziza brassicae on leaves of oilseed rape

Experiments in controlled environments were carried out to determine the effects of temperature and leaf wetness duration on infection of oilseed rape leaves by conidia of the light leaf spot pathogen, Pyrenopeziza brassicae . Visible spore pustules developed on leaves of cv. Bristol inoculated with P. brassicae conidia at temperatures from 4 to 20°C, but not at 24°C; spore pustules developed when the leaf wetness duration after inoculation was longer than or equal to approximately 6 h at 12–20°C, 10 h at 8°C, 16 h at 6°C or 24 h at 4°C. On leaves of cvs. Capricorn or Cobra, light leaf spot symptoms developed at 8 and 16°C when the leaf wetness duration after inoculation was greater than 3 or 24 h, respectively. The latent period (the time period from inoculation to first spore pustules) of P. brassicae on cv. Bristol was, on average, approximately 10 days at 16°C when leaf wetness duration was 24 h, and increased to approximately 12 days as temperature increased to 20°C and to 26 days as temperature decreased to 4°C. At 8°C, an increase in leaf wetness duration from 10 to 72 h decreased the latent period from approximately 25 to 16 days; at 6°C, an increase in leaf wetness duration from 16 to 72 h decreased the latent period from approximately 23 to 17 days. The numbers of conidia produced were greatest at 12–16°C, and decreased as temperature decreased to 8°C or increased to 20°C. At temperatures from 8 to 20°C, an increase in leaf wetness duration from 6 to 24 h increased the production of conidia. There were linear relationships between the number of conidia produced on a leaf and the proportion of the leaf area covered by 'lesions' (both log₁₀-transformed) at different temperatures.     

Spatial and temporal variation of reactive oxygen species and antioxidant enzymes in o-hydroxyethylorutin-treated tomato leaves inoculated with Botrytis cinerea

Pretreatment of detached tomato leaves with o -hydroxyethylorutin reduced the percentage leaf area affected, and delayed the appearance of necrosis, following inoculation with conidial suspensions in droplets of the grey mould fungus Botrytis cinerea . o -Hydroxyethylorutin delayed, but did not inhibit, in vitro germination of conidia, although overall percentage germination was reduced compared with water controls. Both the reactive oxygen species (ROS) – superoxide anions and hydrogen peroxide – increased twice as much in o -hydroxyethylorutin-treated leaf tissue 2 and 6 h postinoculation with B. cinerea conidia in tissues under inoculation drops, as well as in surrounding tissues, whereas in plants not pretreated with the compound ROS generation was noticed later, and only in tissues under inoculation drops. Compared with these compounds, changes in the levels of hydroxyl radicals, lipid peroxidation and the activity of the enzymes superoxide dismutase, guaiacol peroxidase, ascorbate peroxidase and catalase were largely unchanged. In stimulating ROS in inoculated tomato tissue, B. cinerea appeared to be affected directly pre- and postinfection, but indirect effects increasing host resistance cannot be excluded.     

Infection process of Phoma koolunga on stem and leaf tissue of resistant and susceptible field pea (Pisum sativum)

This study investigates the infection process of Phoma koolunga on field pea (Pisum sativum) stems and leaves using different susceptible and resistant pea genotypes for each tissue, viz. 05P778‐BSR‐701 (resistant) and 06P830‐(F5)‐BSR‐5 (susceptible) for stems and ATC 866 (resistant) and ATC 5347 (susceptible) for leaves. On both resistant and susceptible genotypes, light and scanning electron microscopy showed P. koolunga conidia infect stem and leaf tissues directly via appressoria or stomatal penetration, but with more infections involving formation of appressoria on stems than on leaves. On leaves of the resistant genotype, at 72 h post‐inoculation, P. koolunga penetrated more frequently via stomata (5.2 conidia per 36 893 μm²) than by formation of appressoria (1.8 conidia); yet no such difference was observed on stems of the resistant genotype. In contrast, at the same time point, the number of conidia infecting the susceptible genotype by formation of appressoria on either stems (135 conidia) or leaves (11.3 conidia) was significantly greater than via stomata (8 and 7.3 conidia, stems and leaves, respectively). Mean germ tube length of germinating P. koolunga conidia on both stems (29.8 μm) and leaves (32.9 μm) of the resistant genotype was less than on the susceptible genotype (40.5 and 63.7 μm, stem and leaves, respectively). In addition, there were differences in the number of germ tubes emerging from conidia on resistant and susceptible genotypes. These are the first insights into the nature of leaf and stem resistance mechanisms operating in field pea against P. koolunga.     

Peroxidase activity in rice leaves infected withHelminthosporium oryzae in relation to lesion maturation

Two-week-old seedlings of two rice varieties, resistant CH13 and susceptible Benibhog, were inoculated with a conidial suspension ofHelminthosporium oryzae. The resistant host exhibited greater peroxidase activity than the susceptible one when infected with the pathogen. Enzyme activities were measured 0, 24, 48, 72, 96 and 120 h after infection. Peroxidase activity in diseased leaves bearing young lesions,i.e., 24 h after inoculation, was not appreciably different from that in non-infected leaves. At the intermediate and mature stages of lesion development,ri.e., 72 and 120 h after inoculation, a marked increase in peroxidase activity was observed. The pattern of increased enzyme activity was similar in both varieties but the whole event proceeded faster in the resistant variety. No marked change in peroxidase activity in healthy leaves of both the varieties was noticed with increase of seedling age.     

Hypersensitive reaction in alfalfa (Medicago sativa) induced by alfalfa mosaic virus

Alfalfa (Medicago sativa) cv. Beaver was screened for resistance to alfalfa mosaic virus (AIMV). A1MV infection in clonal plants was tested by double-antibody sandwich ELISA 4 and 8 weeks after inoculation with purified A1MV (1 mg/ml in 0025 M phosphate buffer, pH 7 0). Twelve clones showed a hypersensitive reaction 3–4 days after inoculation and the infection was restricted to the inoculated leaves. All the plants of hypersensitive clones consistently produced local lesions when inoculated with purified AIM V. In contrast, plants inoculated with AIMV in crude sap remained symptomless, although AIMV was detectable in inoculated leaves. The remaining 16 clones were susceptible to AIMV and showed systemic infection.