Microtubule dynamics in compatible and incompatible interactions of soybean hypocotyl cells with Phytophthora sojae

The arrangement of microtubules in soybean ( Glycine max ) cells was examined during compatible and incompatible interactions of hypocotyls of soybean cv. Harosoy (susceptible) and cv. Haro 1272 (resistant) with race 1 of the soybean-specific pathogen Phytophthora sojae . Both reaction types were similar during the first 3 h after zoospore inoculation in terms of the number of cells penetrated, and depth penetrated into the cortex. By 3 h postinoculation, clear differences had developed between the two interaction types: incompatible interactions were characterized by a hypersensitive response that was confined to single penetrated cells; while compatibly responding cells appeared unchanged. Both types of response were characterized by autofluorescence of cell walls or cytoplasm and, at 6 h after inoculation, complete disorganization of cell cytoplasm. Reorientation and loss of microtubules was seen in the early stages of the incompatible interaction in association with cellular hypersensitivity, but not in compatible responses. In cells adjacent to those that reacted hypersensitively, there was little evidence of change in microtubule orientation. Treatment of hypocotyls with the microtubule depolymerizer oryzalin prior to inoculation did not alter the compatible response, but led to breakdown of the incompatible response. Changes in microtubule orientation and state are thus among the first structural changes that are visible within cells during incompatibility in this system.     

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.     

A light and scanning-electron microscopic study of infection of tomato plants by virulent and avirulent races of Cladosporium fulvum

Infection of tomato plants byCladosporium fulvum Cooke was studied using light and scanning-electron microscopy. Races 1.2.3 and 4 ofCladosporium fulvum were used, whereas tomato cultivars, carrying the Cf2 gene (susceptible to race 1.2.3 and immune to race 4) and the Cf4 gene (immune to race 1.2.3 and susceptible to race 4) served as differentials. No differences were observed in growth between compatible and incompatible combinations during germination, subsequent formation of runner hyphae and stomatal penetration. Runner hyphae did not show directional growth towards stomata. Penetration usually occurred on the third or fourth day after inoculation. In compatible combinations the fungus grew intercellularly, often in close contact with spongy mesophyll cells. Under optimal conditions it did not cause visible damage to plant cells during early stages of infection. Under suboptimal conditions in winter, the host cells often reacted with callose deposition, but growth of the fungus did not appear to be inhibited. Ten to twelve days after inoculation conidiophores emerged through the stomata and produced conidia. In incompatible combinations fungal growth was arrested one to two days after penetration and confined to stomata and surrounding cells. Very soon the host cells, in contact with the fungus, deposited extensive amounts of callose. Later these cells turned brown and collapsed. At the surface of the host cells, contacted by fungal hyphae, abundant extracellular material could be observed by scanning-electron microscopy. Removing the epidermis of leaves before inoculation delayed the resistant response. On stripped leaves the rate of fungal growth was equal for both interactions up to ten days after inoculation, but the incompatible combination lacked sporulation.     

水稻与稻瘟病菌非亲和性互作中重要防御酶活性变化规律研究

 选以CO39为背景的水稻抗稻瘟病近等基因系,与稻瘟菌生理小种ZC₁₃(菌株97-151a)组成的3类典型非亲和性互作,以亲和性互作为对照,对各互作中过氧化物酶(POD)、苯丙氨酸解氨酶(PAL)、几丁质酶及β-1,3-葡聚糖酶的活性变化规律进行了系统研究。完全非亲和性互作C101A51/97-151a、高度非亲和性互作C101L AC/97-151a及中度非亲和性互作C104 PKT/97-151a,POD比活性接种后即开始明显升高,48h前达到高峰,升高趋势一直持续到7d完全显症时,幅度基本与各互作非亲和程度呈正相关;亲和性互作CO39/97-151a接种后40 h POD比活性才开始升高,4~6 d达到高峰,峰值也较大。3类非亲和性互作PAL比活性在接种后0 h或16 h开始较明显升高,整个互作中形成3~4个较明显的峰;亲和性互作中PAL比活性一直明显下降。3类非亲和性互作外切几丁质酶比活性接种后即开始升高,基本一直保持升高趋势,在40 h前幅度较大,并形成1~3个较高的峰;亲和性互作外切几丁质酶比活性接种后即开始大幅度升高直至完全显症,48h后幅度远高于非亲和性互作。3类非亲和性互作β-1,3-葡聚糖酶比活性在24 h内开始较明显升高,在48h前形成2~3个较明显的峰;亲和性互作在接种后β-1,3-葡聚糖酶比活性即开始升高,在48h后显著高于非亲和性互作。讨论了POD、PAL、几丁质酶及β-1,3-葡聚糖酶参与水稻抗稻瘟病的可能性。     

Ultrastructural Observations and DNA Degradation Analysis of Pepper Leaves Undergoing a Hypersensitive Reaction to Xanthomonas campestris pv. Vesicatoria

Ultrastructural details of the hypersensitive reaction induced by infiltration with avirulent race 2 Xanthomonas campestris pv. vesicatoria in pepper Early Calwonder-10R leaves (incompatible interaction) are reported. Affected cells displayed plasmalemma undulations and disruption, lysis of the chloroplast membrane, degeneration of other organelles, general cytoplasm disorganisation and, often, protoplast shrinkage. The nuclei contained large masses of electron-dense material, apparently formed by chromatin aggregation. In many cases a single chromatin-like layer was deposited on the inner side of the nuclear envelope leaving a finely granular matrix in the centre of the nucleus; the nucleolus usually disappeared. The nuclear envelope was sometimes ruptured and the internal matrix leaked into the cytoplasm. The content of many affected cells eventually coagulated and became very electron-dense. The walls often collapsed. All these alterations were especially visible in spongy mesophyll cells at sites where bacteria occurred in the intercellular spaces. Although some of the nuclear and cytoplasmic alterations recall certain aspects of apoptotic cell death, molecular determinations did not reveal any DNA degradation in hypersensitively reacting tissues. The first cell alterations in leaves infected with the virulent bacterial race 1 (compatible interaction) were observed only 27h after inoculation, when the cytoplasm of some cells showed limited internal disorganisation and plasmolysis at sites where bacterial colonies developed.     

The Effect of Soil Moisture and Cabbage Amendment on the Thermoinactivation of Phytophthora nicotianae

The analysis of the effect of soil water matric potential and temperature regimes on the inactivation of chlamydospores of Phytophthora nicotianae in cabbage amended soils was evaluated using three matric potentials (0, -10, and -30kPa), temperature regimes of 1.5h at 44°C, 5h at 41°C and 8h at 35°C, or 3h at 47°C, 5h at 44°C and 8h at 35°C, with a baseline temperature of 25°C during the rest of the day. The results indicated that survival of P. nicotianae was lowest in saturated soil; and as temperature increased, survival of the pathogen decreased at all soil water matric potentials evaluated. Cabbage amendments can enhance the effect of the heat treatment, further decreasing the pathogen population. The soil water matric potentials evaluated represent optimum levels for the study of thermal inactivation. However, under field conditions lower potentials may be found. Extending the range of soil water matric potentials and the treatment time would allow better comparisons with the field data. There is a clear indication that one irrigation period prior to solarization would provide enough moisture to inactivate the primary inoculum of P. nicotianae in the top soil under field conditions; however, other factors may affect the effectiveness of solarization, reducing or enhancing its potential.     

Study of Defense-related Gene Expression in Grapevine Leaves and Berries Infected with Botrytis cinerea

Defense responses of grapevine towards Botrytis cinerea were investigated. The expression of genes coding for proteins involved in defense were studied: (a) phenylalanine ammonia-lyase (PAL) and stilbene synthase (StSy), (b) an acidic chitinase (VCH3) and a basic chitinase (VCHIT1b), and (c) a polygalacturonase inhibitor protein (PGIP). Since no PGIP was known in grapevine, a complete cDNA sequence was first characterized by PCR and RACE-PCR amplifications. RNAs isolated from infected leaves and infected berries were analysed by semi-quantitative and real-time RT-PCRs. In infected leaves, the expression of PAL, StSy, PGIP and VCH3 genes occurred 6hours post inoculation (hpi). Increase of VCHIT1b gene expression was delayed (24hpi). Maximum levels of induction of these genes were observed at 48hpi, except for the VCH3 gene (24hpi). Activation of these defense responses was not sufficient to stop B. cinerea spread. In berries, no VCH3 gene expression was detected. Maximum levels of induction were observed in stage 3 (loss of berry colour and abundant production of conidia) for the PAL and PGIP genes, and in stage 4 (shrivelled berry) for the StSy and VCHIT1b genes.     

Fine Structure of the Early Interaction of Lily Roots with Fusarium oxysporum f.sp. lilii

The early interaction of lily roots with the cortical rot pathogen Fusarium oxysporum f.sp. lilii was studied using roots of lily bulblets grown in Hoagland's solution, inoculated with the pathogen, and sampled up to 48h later. Conidia produced germ tubes within 6h, which extended towards and into the mucilage covering the root elongation zone, and along and into the anticlinal grooves and middle lamellae of epidermal cells. By 24–48h, infecting hyphae had reached the periclinal walls and intercellular spaces between the epidermis and the outermost cells of the cortex. Penetration of intercellularly growing hyphae directly across host cell walls was not observed; invasion of the cell lumen only occurred by gradual infringing of hyphae upon successive primary wall layers. Non-cellulosic wall appositions rich in vesicles and covered by a cellulosic protective-like layer were formed in response to approaching hyphae in resistant cv.Connecticut King, but rarely in susceptible cv. Esther which seemed more susceptible to plasmolysis and rot. Finger-like projections of the appositions into the host cell cytoplasm likely represent early stages of transfer cell formation.     

Role of Salicylic Acid in Systemic Resistance Induced by Pseudomonas spp. Against Pythium aphanidermatum in Cucumber Roots

Pseudomonas corrugata strain 13 and P. aureofaciens strain 63-28, applied to roots, induced systemic resistance against Pythium aphanidermatum in cucumber roots. Salicylic acid (SA) from bacterial culture or plant tissues was quantified by high performance liquid chromatography. Both strains produced SA in King's B broth and also induced cucumber root to accumulate endogenous SA one day after bacterial inoculation. Using a split root system, more SA accumulated in roots treated with bacteria than in distant roots on the opposite side of the root system in the first two days, but this difference disappeared after 3–4 days. SA levels were significantly higher in plants treated with bacteria compared to the split control, from one to five days after bacterization. SA did not inhibit mycelial growth of Pythium aphanidermatum at 100–200µgml^–1 in vitro, but higher levels inhibited mycelial growth. Zoospore germination increased at concentrations of 10–500µgml^–1, but decreased at 1000µgml^–1 compared to lower concentrations. Exogenously applied SA failed to induce local or systemic resistance against a challenge infection by the pathogen in planta. The results of this study show that exogenous applied SA does not induce systemic resistance to cucumber root rot caused by P. aphanidermatum, but endogenous SA accumulation in cucumber roots may be involved in induced systemic resistance.     

Suppressive effect of potassium silicate on powdery mildew of strawberry in hydroponics

From 1996 to 1997, potassium silicate (SiO₂) was tested at 0, 25, 50, and 100mgl^–1 in hydroponics to control powdery mildew. Other elements were added in the usual amounts, and the strawberries were cultivated hydroponically in a greenhouse for 4 months (from October to January). The powdery mildew spread in the control plot, but little mildew developed in the plot with 25mgl^–1 silicate, and none in plots with more than 50mgl^–1 silicate. The suppressive effect lasted for about 4 months on fruits and even longer on leaves. On analysis of mineral content in the leaves, only the silicate content differed markedly between the control and treated plants. Nitrogen, phosphate, potassium, and calcium contents did not differ greatly. The maximum silicate content was about 24 times that of the control, and disease severity decreased significantly when the content was more than 1.5% in the leaves. The hardness of the strawberry leaves, measured by a creep meter, was increased by the silicate treatment.     

The Role of Trichoderma harzianum Protease in the Biocontrol of Botrytis cinerea

The role of protease of Trichoderma harzianum in the biocontrol of Botrytis cinerea was examined. Two isolates of T. harzianum were compared for their ability to produce protease in liquid culture medium and on the surface of bean leaves. The biocontrol agent T. harziaum T39 produced 58mU/ml of protease and T. harzianum NCIM1185 produced 54mU/ml on the 5th day of growth in liquid culture medium. On bean leaves, combinations of B. cinerea and T. harzianum isolates were examined for the synthesis of protease. The protease activities were 0.9 and 0.6mU/ml for T. harzianum T39 and NCIM1185, respectively, and 0.5mU/ml for B. cinerea alone after 48h of incubation. In the presence of T. harzianum T39 culture liquid containing protease, a 55% reduction in B. cinerea germination and a 80% reduction in the germ tube length were observed after 17h of incubation in vitro. When T. harzianum isolates were added to B. cinerea on bean leaves, increased synthesis of protease was observed (1.0 and 1.2mU/ml for T39 and NCIM1185, respectively). In the presence of T. harzianum NCIM1185 protease, although the rate of germination was reduced, B. cinerea attained 98% germination after 17h of incubation. The hydrolytic enzymes produced by B. cinerea, endo-polygalacturonase (PG) and exoPG were partially deactivated by protease from the T. harzianum isolates. Carboxymethyl cellulase was deactivated only by protease of NCIM1185. On the surface of bean leaves, the protease (obtained from liquid culture medium of T. harzianum isolates) resulted in 56–100% reduction of disease severity. The culture liquid containing protease synthesized on the surface of bean leaves treated with B. cinerea and with T. harzianum was collected and added to fresh leaves infected by B. cinerea. There was 56–100% and 30–75% reduction of disease severity with liquid droplet collected from the leaves treated with T. harzianum T39 and NCIM1185, respectively. Increased control of disease was obtained by combining the conidia of T. harzianum isolates with protease obtained from culture media. Protease inhibitors, trans-epoxysuccinyl-L-leucylamido-(4-guanidino)butane (E64), antipain hydrochloride, and a mixture of inhibitors, but not pepstatin A, fully or partially nullified the biocontrol effect of T39. T39 was found to be a poor producer of chitinase and -1,3-glucanase in vitro. These enzymes were not detected on leaves treated with T39. Involvement of protease in biocontrol of B. cinerea is suggested.     

Parasitoids of Saperda populnea (L.) (Coleoptera: Cerambycidae) on aspen ( Populus tremula L.) in Bulgaria

The parasitoids of Saperda populnea (L.) (Col.: Cerambycidae) were studied in Bulgaria during the period 1997–2001. Galls with pest larvae were collected from young aspen trees at eight locations (Sofia, Kokaliane, Plana, Churek, Gorni Lom, Gintzi, Dolno Kamartzi and Klisura) and examined under laboratory conditions. Four species were reared from 1118 galls containing overwintering pest larvae: Iphiaulax impostor (Scop.) (Hym.: Braconidae), Dolichomitus populneus (Ratz.), Schreineria populnea (Gir.) (Hym.: Ichneumonidae) and Billaea irrorata (Meig.) (Dipt.: Tachinidae). All of the parasitoids were solitary. Schreineria populnea was found as a new parasitoid of the host in Bulgaria. B. irrorata emerged mainly from late-stage S. populnea larvae; the remaining species from both early- and mid-stage larvae. In the parasitoid complex of S. populnea, the most numerous were B. irrorata (59.8%) and D. populneus (29.4%). The percentage of host larval parasitism varied from 2.4 to 33.3, with an average of 9.7%. B. irrorata was the most important in reducing the pest number. The average mortality of S. populnea caused by the tachinid was 5.8%, and the highest mortality observed in individual study was 20.2%.     

Prey consumption during development as well as longevity and reproduction of Typhlodromus pyri Scheuten (Acari, Phytoseiidae) at higher temperatures in the laboratory

The predatory mite Typhlodromus pyri Scheuten (Acari, Phytoseiidae) has been reported as an important predator of the European red mite, Panonychus ulmi (Koch) (Acari, Tetranychidae) in apple culture and vineyards at below 25°C. However, sufficient biological data was lacking on its efficiency at temperatures above 25°C. Therefore, the purpose of the present laboratory work was to obtain experimental data on prey consumption during development as well as longevity and reproduction of T. pyri on apple leaf discs and in Plexiglas cells at constant temperatures of 25±2°C and 30±2°C with P. ulmi as prey.The results showed that mean daily and total prey consumption by both the nymphs and adults of T. pyri decreased significantly on both the arenas as the temperature was increased from 25°C to 30°C, whereby adult prey consumption, both mean daily and total, was higher than that of nymphs. Prey consumption by both the nymphs and adults was significantly higher in the Plexiglas cells than on the leaf discs at both temperatures. Mean total prey consumption during nymphal development was 16.1 () and 12.8 () at 25°C compared to 7.0 () and 5.8 () preys at 30°C on the apple leaf discs and 46.0 () and 38.5 () at 25°C compared to 25.2 () and 20.3 () preys at 30°C in the Plexiglas cells. Mean duration of nymphal development was similar for the two sexes at the same temperature, but it was longer at 25°C than at 30°C. It was 6.0 and 4.0days on the apple leaf discs while 7.0 and 6.0days in the Plexiglas cells at 25°C and 30°C, respectively. Mean daily and total prey consumption by both male and female adults also decreased with the increasing temperature, whereby the females consumed more than double the mean total number of prey than the males on both the arenas of observation and at both temperatures: 355.4 versus 149.7 preys at 25°C and 192.2 versus 85.6preys at 30°C on the leaf discs and 826.8 versus 374.5 preys at 25°C and 488.9 versus 187.9 preys at 30°C in the Plexiglas cells. Longevity of the females was longer than males on both arenas and at both temperatures and it was longer at 25°C than at 30°C. Mean total longevity on the apple leaf discs was 68.3 () and 50.8 () days at 25°C compared to 52.5 () and 36.8 () days at 30°C, while in the Plexiglas cells it was 91.0 () and 65.8 () days at 25°C compared to 75.3 () and 48.5 () days at 30°C. Reproduction in females also decreased significantly with increasing temperature. It decreased from 62.0 to 39.0 eggs/female on the leaf discs and 75.0 to 47.1 eggs/female in the Plexiglas cells. The females laid significantly higher numbers of eggs at both temperatures in the Plexiglas cells than on the leaf discs. Oviposition period in females was 30days at 25°C on both the arenas, while at 30°C it was 26days on the apple leaf discs and 27days in the Plexiglas cells.     

Influences of host age, sex ratio, population density, and photoperiod on parasitism by Trichogramma evanescens Westw. (Hym., Trichogrammatidae)

Moth species Ephestia kuehniella and Sitotroga cerealella are serious pests in cereal-based food processing facilities and stores in Turkey. Control of these pests is undertaken by regular space treatment of infested areas with pesticides. An alternative control method could be the release of parasitic wasps of the genus Trichogramma. In laboratory tests, we use T. evanescens as a parasitic wasp reared on the eggs of Ephestia kuehniella. Adult wasps emerging from the host were maintained in glass tubes at 27±1° C, 60–70±5% r.h and L14:D10 and fed on honey solution. Adults of T. evanescens in vials without food enclosed from host eggs and all died within 1.8days; in comparison, in vials with honey, live adults were evident for 15.2days. Fresh (6–48h) and old (72–96h) host eggs were offered to T. evanescens and fresh eggs were more accepted than old. Host acceptance of females with males in vials was better than without males. The number of females in the vials also influenced the parasitization rate and single female parasitized more eggs. Adults living in light regime (L14:D10 and L6:D18) parasitized more eggs than in total darkness. Results are discussed with relation to enhancing parasitoid effectiveness in biological control in processing facilities and mills.     

Influence of mixed biocide GCSC-BtA on the pupae and adult stages of Apanteles plutellae Kurd. (Hym., Braconidae) and its host. Plutella xylostella (L.) (Lep., Plutellidae)

This paper deals with the influence of the mixed biocide GCSC-BtA on the pupal and adult stages of Apanteles plutellae Kurd. (Hym., Braconidae) and its host, Plutella xylostella (L.) (Lep., Plutellidae). The results show that mortalities of the pupae of P. xylostella in the direct-dip bioassay were 84.67%, that of the adults in the residue bioassay at 1.2500mg/ml concentration of GCSC-BtA were 78.00% which were significantly higher than the mortality values for the pupae with 54.62% and adults with 48.13% of A. plutellae. In contrast, cypermethrin showed extremely high toxicity to the pupae with 94.58% and adults with 86.00% mortality values of A. plutellae as compared to the low mortality values of 42.14% for the pupae and 32.11% for the adults of P. xylostella, with the same concentrations and bioassay methods. The LC₅₀ values of GCSC-BtA were 0.3402, 0.5516 and 1.2405, 1.9480mg/ml for the pupae and adults of P. xylostella and A. plutellae, respectively, while the LC₅₀ values for cypermethrin were 1.5652, 2.3471 and 0.1096, 0.1152mg/ml, respectively. GCSC-BtA was found more toxic to the pupae and adults of P. xylostella and safer to the pupae and adults of A. plutellae than cypermethrin. The possibilty of using GCSC-BtA against P. xylostella under partial control by A. plutellae in vegetable fields is discussed.     

Germination and invasion by ascospores and pycnidiospores of <Emphasis Type="Italic">Leptosphaeria maculans</Emphasis> on spring-type <Emphasis Type="Italic">Brassica napus</Emphasis> canola varieties with varying susceptibility to blackleg

The infection processes of ascospores and pycnidiospores of Leptosphaeria maculans were studied on cotyledons of six cultivars of spring-type Brassica napus: one with resistance controlled by a single dominant gene (cv. Surpass 400), three with polygenic resistance (cvs. Dunkeld, Grouse, and Outback), and two susceptible cultivars (Westar and Q2). On all cultivars, ascospore germination, penetration, and development of symptoms on cotyledons were much earlier than that with pycnidiospores. At 2h after inoculation ascospores began to germinate, by 4h about 50% had germinated, and by 6–8h 85%–90% had germinated. In contrast, pycnidiospores began to germinate 1 day after inoculation (dai) and reached only 50% germination by 3 dai. Ascospores began germinating from terminal cells and then later from the interstitial cells. Pycnidiospores germinated predominantly from one end and sometimes from both ends. Germ tubes from ascospores penetrated stomata as early as 4h after inoculation, whereas those from pycnidiospores penetrated at 2 dai. Symptom development with ascospores was 2 days earlier than that with pycnidiospores. Symptoms on Surpass 400 were evident as early as 3–5 dai with ascospores and 5–7 dai with pycnidiospores. However, on other cultivars, symptoms were not evident until 10 dai with ascospores and 12 dai with pycnidiospores. This report is the first on differences in the infection processes by the two spore types. Ascospore and pycnidiospore attachment, germination, and penetration did not differ between resistant and susceptible cultivars, but there were major differences after penetration. Under high humidity, 80%–90% of stomata of susceptible Westar and Q2 had aerial hyphae emerging from stomatal pores. However, fewer stomata (5%–10%) had aerial hyphae on Surpass 400 by 10 dai with ascospores and 12 dai with pycnidiospores, but even these were usually poorly developed. Host differences in spring-type B. napus in relation to production of aerial hyphae have not previously been reported. In Surpass 400, rapid necrosis of guard cells occurred within a few hours of penetration by either type of spore, and subsequently one or a few cells immediately adjacent to the penetration site died. This necrosis then spread to the cells around the penetration site to form a hypersensitive response (in the form of a small, dark lesion) to both ascospores and pycnidiospores. This is the first detailed report on interactions between spring-type B. napus and L. maculans in relation to single dominant gene-based resistance. Neither the cultivars with polygenic resistance nor the susceptible cultivars had such a response.     

白叶枯病菌与非亲和水稻IRBB4悬浮细胞互作中蛋白类激发子的纯化和鉴定

 本研究对水稻白叶枯病菌与水稻悬浮细胞非亲和互作中蛋白类激发子进行了分离纯化和鉴定.白叶枯病菌JXOV与水稻IRBB4和IR24悬浮细胞互作36 h后的上清液,经Q-Sepharose阴离子交换层析柱分离,对分离的各组分进行抗病性诱导测定,结果表明JXOV与IRBB4非亲和互作的上清液中存在蛋白类激发子.有活性的蛋白组分经阴离子交换层析柱Mono-Q进一步纯化后,SDS-PAGE分析鉴定出2个具激发活性的蛋白,其分子量分别为17.2 kD和49.2 kD,等电点分别为5.8和6.2.利用上述激发子处理水稻能减少病斑长度并诱导水稻防卫酶活性的增加.     

Analysis of Defense-related Proteins in Stem Tissue of Carnation Inoculated with a Virulent and Avirulent Race of Fusarium oxysporum F.sp. Dianthi

The aim of this study was to learn more about the accumulation of defense-related proteins in stem tissue from carnation cultivar Pallas inoculated with 2 near-isogenic races, the avirulent race 1 and the virulent race 8 of Fusarium oxysporum f.sp. dianthi. Stem tissue was used, from which the epidermis, cortex and medulla were peeled off from the vascular cylinder. It appeared that chitinase activity was constitutively expressed in the intercellular fluids (IFs) of untreated leaves, stems and roots of carnation. The total chitinase activity in the IFs of stem tissue increased with time after inoculation. This increase was similar after inoculation with the virulent, the avirulent race and water. At least four chitinase isoenzymes, three acidic and one basic isoform, were detected in the IFs of inoculated plants. In contrast, total 1,3--glucanase activity was not detected in the IFs of untreated leaves, stems and roots. Furthermore, the increases in 1,3--glucanase activity in IFs of stem tissue were markedly higher in the compatible and incompatible interactions than in the water control, indicating that this activity is specially induced by elicitors common to both races 1 and 8 of Fusarium oxysporum f.sp. dianthi. Using an antiserum against 1,3--glucanase P3 of tomato, 2 bands were detected on immunoblots in the IFs of stem tissue inoculated with races 1 and 8. No bands were visible after inoculation with water. Total peroxidase activity increased with time in all combinations. One basic and one acidic peroxidase isoform were present in these IFs. Peroxidase activity in a cell wall fraction prepared from stem tissue was clearly higher, and its increase faster, than the activity in the soluble stem fraction. These increases were similar in the virulent, the avirulent race and the water control. The growth of the fungus Trichoderma viride was inhibited by the IFs obtained from stem tissue inoculated with the virulent and the avirulent race of Fusarium oxysporum f.sp. dianthi. However, the growth of Fusarium oxysporum f.sp. dianthi itself was not affected by these IFs.     

Infection of Linseed by Alternaria linicola; Effects of Inoculum Density, Temperature, Leaf Wetness and Light Regime

Controlled environment studies were conducted to determine the effects of inoculum density, temperature, leaf wetness and light regime on the infection of linseed by Alternaria linicola. The % cotyledons and leaves with symptoms, and the disease severity (% leaf area with symptoms) increased linearly when the inoculum density increased from 1×10³ to 1×10⁵ conidiaml^–1. The first symptoms appeared on cotyledons and leaves 4 and 6 days after inoculation, respectively. Eight hours of leaf wetness were sufficient to initiate the disease at 25°C but not at 15°C, when 10-h periods of leaf wetness were required. % leaf area with symptoms was lower at 15°C than that at 25°C irrespective of the leaf wetness periods tested. Interruption of a continuous leaf wetness period by a 12-h dry period, occurring at any time between 1 and 18h after inoculation, decreased the % cotyledons with symptoms and the disease severity, with the greatest reductions (60% and 100%, respectively) being observed when the dry period began 6h after inoculation. A. linicola conidia were able to exploit successive 12-h periods of leaf wetness cumulatively to infect linseed plants. Disease incidence and severity were positively correlated with the dark period following inoculation, but they were negatively related to the length of the initial light period. Our findings suggest that infection of linseed by A. linicola and further development of symptoms can occur under unfavourable environmental conditions.