叶锈病侵染对大麦碳素代谢的影响

 本研究用¹⁴C同位素示踪法测定叶锈病侵染引起的大麦幼苗光合作用和光合产物分布的病变。试验以植株第一叶和第二叶所得的结果基本相同:接菌7天后病叶总光合量逐渐降低(10天后降低到健叶的60%以下)而单位绿叶面积光合率则明显提高(10天后提高约20%),且这种光合病变与锈病严重度间呈密切的线性相关(r ≥ 0.949,P<0.001);接菌3天后病叶光合产物的输出明显受阻,其截留的¹⁴C物质大量增加(为对照的3倍多)而输运到根和茎部的¹⁴C物质却严重减少。这些病变无疑是叶锈病抑制大麦植物生长发育的主要生理原因之一。     

山定子抗苹果褐斑病菌侵染过程中DNA ladder与类caspases活性的检测

接种褐斑病菌分生孢子悬浮液于山定子与富士苹果叶片,接种后1、3 d和5 d取样,提取DNA,检测DNA ladder;提取样品粗蛋白,用特异性荧光底物检测粗蛋白中类caspase活性,探索山定子受褐斑病菌侵染过程中引起的细胞程序性死亡(PCD)与抗性的关系,检测DNA ladder和类caspases活性变化的规律。研究发现:在接种后1、3 d和5 d,山定子和富士苹果叶片中没有明显DNA ladder的产生;受检测的YVADase、DEVDase、IETDase和VEIDase活性在接种后1 d和3 d没有显著变化,但在接种后5 d,活性均降低30%左右,显著低于对照。研究表明,山定子在褐斑病菌侵染过程中并没有DNA ladder产生,但却伴随着类caspase活性的下降。     

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.     

Effects of wetness period and temperature on development of dark pod spot (Alternaria brassicae) on oilseed rape (Brassica napus)

In controlled environment experiments, when oilseed rape pods or leaves were inoculated with spore suspensions of Alternaria brassicae, the maximum disease incidence (proportion of pods or leaves diseased) increased as wetness period after inoculation increased from 4 to 24 h and as temperature increased to 20°C. There was a clear relationship between disease incidence on pods and incidence on leaves with the same wetness/temperature conditions. Logistic equations described the effects of wetness period after inoculation on disease incidence (number of pods or leaves infected) or disease severity (number of lesions on pods or leaves) using temperature-dependent and tissue-dependent parameters. The time from inoculation to the appearance of the first lesions was shorter on pods than on leaves at temperatures ≤15°C and wetness periods ≤12 h. Two-dimensional response surface equations or simple interpolations from one-dimensional equations were used to develop contour maps of expected disease incidence and severity, respectively, on leaves or pods to estimate the effects of different combinations of wetness period during infection and temperature on disease development.     

Quantitative detection and monitoring of Colletotrichum acutatum in strawberry leaves using real-time PCR

Real-time PCR assays for Colletotrichum acutatum , one of the most important pathogens of strawberry worldwide, were developed using primers designed to the ribosomal DNA internal transcribed spacer 1 (rDNA ITS1) and the β-tubulin 2 gene. Using TaqMan technology, the ITS-based assay could reliably detect as little as 50 fg genomic DNA, 100 copies of target DNA, or 25 conidia. The β-tubulin-based assay was c . 66 times less sensitive, and therefore less suitable for detection purposes. The TaqMan-ITS assay recognized all C. acutatum isolates tested from various intraspecific molecular groups, while no amplification was observed with several other Colletotrichum species or other strawberry pathogens, indicating the specificity of this assay. Detection and quantification of C. acutatum was demonstrated in artificially and naturally infected strawberry leaves. First, C. acutatum was detected in plant mixes of which only 0·001% of the tissue was infected by C. acutatum . Secondly, real-time PCR analysis of leaf samples taken at various times after inoculation indicated that the assay allowed monitoring of growth progression of C. acutatum . This real-time PCR-mediated monitoring of the pathogen was well-correlated with microscopic data, and confirmed that leaf age may play a role in the extent of C. acutatum infection. Finally, the assay allowed detection of C. acutatum in naturally infected and symptomless strawberry leaves collected from production fields and planting material.     

Alteration of photosynthetic performance and source–sink relationships in wheat plants infected by Pyricularia oryzae

Two experiments were carried out to assess the changes associated with photoassimilate production and partitioning in the source–sink relationship of flag leaves and spikes of wheat plants infected with Pyricularia oryzae, the causal agent of blast. Flag leaves and spikes were inoculated at 10 and 20 days after anthesis (daa) with a conidial suspension of P. oryzae. Analysis of chlorophyll a fluorescence using maximal photosystem II quantum efficiency (F_v?F_m), fraction of energy absorbed that is used in photochemistry (YII), quantum yield of non‐regulated energy dissipation (Y(NO)) and quantum yield of regulated energy dissipation (Y(NPQ)), showed an impairment of the photosynthetic performance in both infected flag leaves and spikes, coupled with reduced concentrations of chlorophyll a  + b and carotenoids. Compared to non‐inoculated controls, there was lower capacity for CO₂ fixation by RuBisCO in the infected flag leaves. Similarly, in the infected flag leaves and grains (obtained from infected spikes), there were lower concentrations of soluble sugars, while the hexoses‐to‐sucrose ratio increased in infected flag leaves. Compared to non‐inoculated controls, infected flag leaves showed lower sucrose phosphate synthase (SPS) activity and lower expression of the sucrose synthesis (SuSy) gene, while higher expression and activity of acid invertases also occurred. At the advanced stages of fungal infection, the concentration of starch in grains decreased but remained high for the infected flag leaves. There were reductions in ADP‐glucose pyrophosphorylase activity and the expression of ADP‐glucose pyrophosphorylase genes and a down‐regulation of β‐ and α‐amylase expression at the advanced stages of fungal infection on flag leaves and spikes. In conclusion, the effect of blast on both grain quality and yield can be associated with alterations in both production and partitioning of carbohydrates during the grain filling process.     

TeMV与PWV侵染对西番莲代谢生理及组织微结构的影响

以西番莲品种‘台农1号’为实验材料,通过电镜扫描技术比较观测TeMV和PWV侵染与健康西番莲的叶片、幼嫩果实外果皮组织形态结构,分析测定其叶片生理生化变化,研究西番莲病毒侵染对西番莲叶片和幼嫩果实外果皮组织结构和生理生化的影响。结果表明：TeMV与PWV侵染后,西番莲植株叶片的海绵组织细胞间隙缩小,发生成团堆积状排列,栅栏组织细胞萎缩,呈不规则长条形;幼嫩果实外表皮组织中石细胞层发生严重断裂,排列松散,海绵层细胞变得松散,且发生较严重的木质化现象;叶片与幼嫩果实外表皮表面纹理粗糙、褶皱,气孔保卫细胞萎缩,气孔周围组织皱缩、粗燥;感病果实外果皮表面上的腺毛凸起无序。病毒侵染后,感病植株第1～3叶的蔗糖含量较第4～6叶的高23.11%;健康叶片则相对平衡,降低了淀粉积累、PG酶、纤维素酶与PPO酶活性,提高了感病植株第1～3叶的α-淀粉酶、GS酶与POD酶活性。西番莲受TeMV与PWV侵染后,植株叶片第1～6叶营养物质分配不平衡,相关物质的合成分解与抗氧化防御酶系统受伤害,叶片与果实组织形态结构受到不同程度的损坏,植株正常吸收功能遭到破坏。     

Effects of fludioxonil and pyrimethanil, two fungicides used against Botrytis cinerea, on carbohydrate physiology in Vitis vinifera L

In Vitis vinifera L, photosynthesis and photosynthate partitioning are affected in the presence of fludioxonil and pyrimethanil, two fungicides commonly used in vineyards against Botrytis cinerea Pers. However, the effects were found to be different according to the model studied: plantlets (cv Chardonnay) grown in vitro, fruiting cuttings (cv Chardonnay) and plants grown in vineyards (cvs Chardonnay, Pinot noir and Pinot Meunier). In the plantlets grown in vitro, both fungicides decreased gas exchanges, photosynthetic pigment and starch concentrations in the leaves, whereas soluble carbohydrates transiently accumulated, suggesting that plantlets mobilised starch in response to photosynthesis inhibition caused by fungicides. In the fruiting cuttings, the fungicides did not affect photosynthesis, although fludioxonil caused starch decrease in parallel with sucrose accumulation, suggesting that the fungicide effects were of lower intensity than in vitro. Conversely, in vineyard, the two fungicides stimulated photosynthesis and increased pigment concentrations in the three vine cultivars tested. In the meantime, glucose, fructose and starch levels of the leaves declined after fungicide exposure, whereas sucrose accumulated, indicating that sucrose synthesis increased in the leaves following the fungicide treatment. Among the three varieties, Chardonnay was the most sensitive to the fungicides as revealed by the intensity of the responses and the longer period for recovery. In vineyard, the results suggested that the two fungicides, in addition to inhibiting B cinerea development, had a beneficial effect on vine physiology through the stimulation of leaf carbon nutrition, which may further enable the plant to rapidly make use of its defence reactions.     

条锈菌侵染早期小麦叶片RNA和rRNA的合成

 采用示踪方法研究了条锈菌侵染早期小麦叶片的RNA和rRNA合成。结果表21h期间,表现不亲和性反应的叶片总RNA合成有所增加,而亲和性反应寄主叶片则低rRNA合成未受侵染影响。在接种后39~45h期间,只有呈亲和性反应的寄主叶片RNA和大幅度增加。对rRNA各组分合成的测定表明条锈菌侵染所影响的只是大分子量rRNA,胞质和叶绿体rRNA的合成也存在差别。文中讨论了上述变化的病理学意义。     

Verticillium dahliae Modifies the Concentrations of Proline, Soluble Sugars, Starch, Soluble Protein and Abscisic Acid in Pepper Plants

The objective of this research was to study the levels of some organic solutes, such as total protein, total soluble sugars, starch and proline in leaves, as well as abscisic acid concentration in xylem of pepper plants inoculated with Verticillium dahliae Kleb. Healthy and inoculated plants were always kept well watered. Measurements were made at time intervals after inoculation. Leaf water potential rapidly decreased as a consequence of fungal infection. However, relative water content in leaves only changed significantly from day 20 after inoculation, and such decreases coincided with a sharp build up of proline and total soluble sugars in leaves. Starch and protein levels, as well as abscisic acid concentration in xylem, declined in healthy and inoculated peppers as they became older. However, such decreases were more pronounced in infected plants, especially soon after inoculation. Results suggest that proline and total soluble sugars accumulation could be sensors of the damage caused by the fungal infection.     

黄瓜白粉病菌拮抗真菌鉴定及生防效果评价

本文对黄瓜白粉病菌Sphaerotheca fuliginea拮抗真菌进行了筛选和鉴定及防治效果的评价。结果表较高的拮抗活性。菌株JK-13发酵液处理白粉菌孢子48 h后,其对孢子萌发抑制率为89.93%;发酵液处理发病的离体叶片和盆栽幼苗7 d后,其防治效果分别为88.02%和76.34%。利用形态学和分子生物学技术鉴定该菌为腐皮镰刀菌Fusarium solani。     

Limited movement of Cucumber mosaic virus (CMV) in yellow passion flower in Brazil

Symptoms of Cucumber mosaic virus (CMV) on yellow passion flower ( Passiflora edulis f. flavicarpa ) are characterized by bright yellow mottling on leaves, starting at random points on the vine and diminishing in intensity towards the tip, which becomes symptomless as it grows. To determine whether symptomless portions of vines are CMV-free or represent latent infection, leaves with and without symptoms were collected from infected vines in the field. Biological, serological (plate-trapped antigen enzyme-linked immunosorbent assay, PTA-ELISA), Western blot and dot-blot hybridization assays showed that portions of the vines without symptoms were CMV-free. Vegetatively propagated vines with symptoms showed remission of symptoms on newly developed leaves. One year later, no CMV was detected in the upper leaves of these plants. Mechanically inoculated passion flower seedlings behaved similarly; symptoms were shown by few leaves after inoculation. Afterwards, plants became symptomless and CMV was not detected in the upper leaves or root system, 40 or 85 days after inoculation. The mechanism responsible for remission of symptoms accompanied by CMV disappearance is not known.     

香蕉束顶病毒(BBTV)侵染对寄主内源激素的影响

 本试验用ELISA方法测定了香蕉感染束顶病毒(BBTV)后植株体内的3种内源激素,赤霉素(GA₃)、玉米素类(iPAs)和脱落酸(ABA)的变化。结果表明,感株的GA₃水平在侵染过程中虽有微弱增加,但含量和增长速度显著低于健株对照;iPAs在接种BBTV第14天后明显下降,并维持较低水平;ABA在BBTV侵染后被大量诱导增加并不断积累,在接种后第35天测定含量最高,为对照的3.34倍。试验还同时检测了BBTV在侵染过程中的运转。用间接ELISA测定的接种叶和顶叶的BBTV含量显示:接种21天后BBTV在接种叶和顶叶中还大量增殖,呈系统性分布。但寄主的症状在接种35天后才逐渐在顶叶表现。以上结果表明:香蕉束顶病的症状表现似乎主要与病株中的内源激素的失调有关,而与BBTV在体内的运转并不直接相关     

First report of Pseudomonas tolaasii on Agaricus bisporus in Serbia

Symptoms of brown blotch were observed on cultivated Agaricus bisporus on several mushroom farms in Serbia. Subsequently, samples were collected from nine localities during 2006?C2010 and fluorescent Gram-negative bacterial strains were isolated. All strains caused sunken brown lesions on A. bisporus tissue blocks after artificial inoculation. Isolated strains showed the LOPAT characteristics of the group Va being negative for levan production; the ability to produce a hypersensitivity reaction in tobacco; were oxidase and arginine dihydrolase positive; and lacked pectolytic activity. The results from other tests were as follows: oxidative metabolism of glucose; catalase activity positive; aesculin hydrolisis negative; Tween 80 hydrolysis positive; casein hydrolysis positive; gelatine hydrolysis and nitrate reduction negative; mannitol, erythritol, sorbitol, inositol and trehalose were used as a carbon source but not sucrose, arabinose and D(-) tartrate. Based on these features bacterial strains were identified as Pseudomonas tolaasii. The identity of isolated strains was confirmed by polymerase chain reaction. This is the first report of P. tolaasii in Serbia.     

The enzymes of glycogen and trehalose catabolism from Hysterothylacium aduncum (Nematoda: Anisakidae)

The activity of alpha-amylase, glucoamylase, maltase, trehalase, glycogen phosphorylase and trehalose phosphorylase was measured in extracts from larval and adult Hysterothylacium aduncum (Rudolphi, 1802), parasitic nematode of marine fish. The content of glycogen and trehalose in the worm's body was also determined. Both the hydrolytic and phosphorolytic paths of sugar decomposition are present in H. aduncum. In the larvae glycogen was utilised mainly via the hydrolytic path. In the adults the activities of phosphorolytic enzymes were higher than in the larvae. In both stages the activity of trehalose phosphorylase is present. In adult nematodes it is uncommonly high. The dominating sugars in the adults were glucose and glycogen, while in the larvae it was trehalose.     

Molecular cloning and expression analysis of genes related to phosphatidic acid synthesis in Japanese pear leaves inoculated with <Emphasis Type="Italic">Venturia nashicola</Emphasis>

Phosphatidic acid, which can be generated by the action of phospholipase D (PLD) and by the combined action of phospholipase C (PLC) and diacylglycerol kinase (DGK), is a plant defense signal. To identify the role of the PLD, PLC, and DGK genes encoding these enzymes in pear resistance to the scab fungus Venturia nashicola, we report the cloning of these genes and analysis of their expression in inoculated pear leaves. The results showed that PLD mRNA increased in the leaves of the susceptible pear cv. Kousui 1 day after inoculation (dai), then decreased to the basal level. In resistant pear cv. Kinchaku, PLD mRNA did not change significantly except for a small increase at 2 and 3 dai. However, the level of PLD mRNA in Kinchaku was higher than in Kousui 2 dai. The level of PLC mRNA significantly increased in cvs. Kousui and Kinchaku by 1 dai, peaked at day 2, then decreased to the basal level. Thus, the gene expression pattern did not differ between the two cultivars. PLC gene expression was also induced by environmental stress. The DGK gene seemed to be constitutively expressed and was not induced by inoculation in either the susceptible or resistant cultivars. Together, this data showed that both PLD and PLC are possibly involved in a defense response to V. nashicola in Japanese pear leaves, and the PLD gene seems to play a more important role than does the PLC gene.     

Comparative microscopic and molecular analysis of Thatcher near‐isogenic lines with wheat leaf rust resistance genes Lr2a,Lr3, LrB or Lr9 upon challenge with different Puccinia triticina races

Thatcher near‐isogenic lines (NILs) of wheat carrying resistance gene Lr2a, Lr3, LrB or Lr9 were inoculated with Puccinia triticina races of virulence phenotype BBBD, MBDS, SBDG and FBDJ. Puccinia triticina infection structures were analysed under the fluorescence microscope over a course of 14 days after inoculation (dai). The relative proportion of P. triticina and wheat genomic DNA in infected leaves was estimated with a semiquantitative multiplex PCR analysis using P. triticina‐ and wheat‐specific primers. The occurrence of a hypersensitive response (HR), cellular lignification and callose deposition in inoculated plants was investigated microscopically. In interactions producing highly resistant infection type (IT) ‘0;’, a maximum of two haustorial mother cells per infection site were produced, and there was no increase in the proportion of P.  triticina genomic DNA in infected leaves, indicating the absence of P. triticina growth. In comparison, sizes of P. triticina colonies increased gradually in interactions producing moderately resistant IT ‘1’ and ‘2’, with the highest proportion of P. triticina genomic DNA found in leaves sampled at 14 dai. In interactions producing susceptible IT ‘3–4’, the highest proportion of P. triticina genomic DNA was found in leaves sampled at 10 dai (45·5–51·5%). HR and cellular lignification were induced in interactions producing IT ‘0;’ and ‘1’ at 1 dai but they were not observed in interactions producing IT ‘2’ until 2 dai. No HR or cellular lignification were induced in interactions producing susceptible IT ‘3–4’. Furthermore, a strong deposition of callose was induced in Lr9 + BBBD and Lr9 + FBDJ (IT ‘0;’), whereas this defence response was not induced in resistant or susceptible interactions involving Lr2a, Lr3 or LrB, indicating that Lr9 mediated resistance was different from that conditioned by Lr2a, Lr3 or LrB.     

The possibility of factors other than phytoalexin accumulation preventing fungal growth in the incompatible interactions of soybean with Phytophthora megasperma f. sp. glycinea

Soybean ( Glycine max ) cv. Harosoy 63 is resistant to race 1 and susceptible to race 9 of Phytophthora megasperma f. sp. glycinea (Pmg). In detached primary leaves inoculated with zoospores, growth of race 1 was completely suppressed 16 h after inoculation, while race 9 was unaffected. The amount of the phytoalexin glyceollin that accumulated, however, was not significantly different in either the incompatible or compatible interaction 16 h after inoculation. At the circumference of the inoculated area, a slight accumulation of phytoalexin was observed only in the incompatible interaction 20 h or more after inoculation. Tolerance of race 9 to the phytoalexin was significantly higher than that of race 1 when the phytoalexin was added to agar. Moreover, race 9 degraded glyceollin faster than race 1. On leaves inoculated at separate points with either race, the lesion associated with race 9 never colonized areas inoculated with race 1. These results suggest that factor(s) other than the accumulation of phytoalexin in soybean tissue might cause cessation of growth of Pmg.