The effect of methyl jasmonate on enzyme activities in wheat genotypes infected by the crown and root rot pathogen <Emphasis Type="Italic">Fusarium culmorum</Emphasis>

The hemibiotrophic pathogen Fusarium culmorum (Fc) causes crown and root rot (CRR) in wheat. In this study, MeJA treatment was done 6 h after pathogen inoculation (hai) to focus the physiological and biochemical responses in root tissue of the susceptible wheat cv Falat, partially resistant cv Pishtaz and the tolerant cv Sumai3 at the beginning of the necrotrophic stage. The results indicate that treatment with MeJA at 6 hai significantly delayed the necrotic progress in cv Falat, whereas no significant difference was seen in other cultivars. The activities of pathogen responsive defense-related enzymes (SOD, CAT, POX, PPO, LOX and PAL), total phenols and callose contents were higher in Sumai3, while treatment with MeJA significantly increased these enzymes activities and total phenols content in Falat, signifying the most sensitive cultivar which had a weak reaction to the pathogen but a strong response to MeJA treatment. Additionally, MeJA treatment decreased the level of H₂O₂ and MDA contents particularly in cv Falat. This is the first work reporting the regulation of defense-related enzymes by MeJA treatment at particular time point of 6 hai suggests the possible role of JA in regulating basal resistance in CRR pathogen–wheat interaction. Taken together, our data add new insights into the mechanism of wheat defense including enzymatic events controlling wheat protection against Fc infection.     

Low-Temperature Effects on Photosynthesis and Correlation with Freezing Tolerance in Spring and Winter Cultivars of Wheat and Rye

Winter cultivars of rye (Secale cereale L., cv Musketeer) and wheat (Triticum aestivum L. cvs Kharkov and Monopol), but not a spring cultivar of wheat (Glenlea), grown at cold-hardening temperatures showed, at high irradiances, a higher proportion of oxidized to reduced primary, stable quinone receptor (QA) than did the same cultivars grown under nonhardening conditions. In addition, there was a positive correlation between the effects of low-growth temperature on this increased proportion of oxidized QA, and a concomitant increase in the capacity for photosynthesis, and LT50, the temperature at which 50% of the seedlings are killed, in cultivars showing different freezing tolerances. This suggests that low-temperature modulation of the photosynthetic apparatus may be an important factor during the induction of freezing resistance in cereals. Finally, the control of photosystem II photochemistry by nonphotochemical quenching of excitation energy was identical for nonhardened and cold-hardened winter rye. However, examination of measuring temperature effects per se revealed that, irrespective of growth temperature, nonphotochemical quenching exerted a stronger control on photosystem II photochemistry at 10[deg] C rather than at 20[deg] C.     

The barley apoptosis suppressor homologue BAX inhibitor-1 compromises nonhost penetration resistance of barley to the inappropriate pathogen Blumeria graminis f. sp. tritici

BAX inhibitor-1 (BI-1) proteins have been characterized as suppressors of programmed cell death in mammals and plants. The barley BI-1 is a suppressor of nonspecific background resistance and mlo-mediated penetration resistance to the biotrophic fungal pathogen Blumeria graminis f. sp. hordei when overexpressed in epidermal cells of barley. We report here that BI-1 expression is also slightly up-regulated during interaction with the inappropriate wheat pathogen Blumeria graminis f. sp. tritici. Significantly, overexpression of BI-1 in single epidermal cells of barley by microprojectile-mediated transformation rendered cells susceptible to penetration by inappropriate B. graminis f. sp. tritici. The degree of transgene-induced accessibility to B. graminis f. sp. tritici was thereby similar to the effect achieved by overexpression of the defense suppressor gene Mlo and could not be further enhanced by double expression of both BI-1 and Mlo. Confocal laser scanning microscopy was used to locate a functional green fluorescing GFP:BI-1 fusion protein in endomembranes and the nuclear envelope of barley epidermal cells. Together, enhanced expression of barley BI-1 suppresses penetration resistance to B. graminis f. sp. tritici, linking barley nonhost resistance with cell death regulation.     

Sucrose metabolism in spring and winter wheat in response to high irradiance, cold stress and cold acclimation

Effects of low‐temperature stress, cold acclimation and growth at high irradiance in a spring (Triticum aestivum L. cv. Katepwa) and a winter wheat (Triticum aestivum L. cv. Monopol) were examined in leaves and crowns with respect to the sucrose utilisation and carbon allocation. Light‐saturated and carbon dioxide (CO₂)‐saturated rates of CO₂ assimilation were decreased by 50% in cold‐stressed spring and winter wheat cultivars. Cold‐ or high light‐acclimated Katepwa spring wheat maintained light‐saturated rates of CO₂ assimilation comparable to those of control spring wheat. In contrast, cold‐ or high light‐acclimated winter wheat maintained higher light and CO₂‐saturated rates of CO₂ assimilation than non‐acclimated controls. In leaves, during either cold stress, cold acclimation or acclimation to high irradiance, the sucrose/starch ratio increased by 5‐ to 10‐fold and neutral invertase activity increased by 2‐ to 2.5‐fold in both the spring and the winter wheat. In contrast, Monopol winter wheat, but not Katepwa spring wheat, exhibited a 3‐fold increase in leaf sucrose phosphate synthase (SPS) activity, a 4‐fold increase in sucrose:sucrose fructosyl transferase activity and a 6.6‐fold increase in acid invertase upon cold acclimation. Although leaves of cold‐stressed and high light‐grown spring and winter wheat showed 2.3‐ to 7‐fold higher sucrose levels than controls, these plants exhibited a limited capacity to adjust either sucrose phosphate synthase or sucrose synthase activity (SS[s]). In addition, the acclimation to high light resulted in a 23–31% lower starch abundance and no changes at the level of fructan accumulation in leaves of either winter or spring wheat when compared with controls. However, high light‐acclimated winter wheat exhibited a 1.8‐fold higher neutral invertase activity and high light‐acclimated spring wheat exhibited an induction of SS(d) activity when compared with controls. Crowns of Monopol showed higher fructan accumulation than Katepwa upon cold and high light acclimation. We suggest that the differential adjustment of CO₂‐saturated rates of CO₂ assimilation upon cold acclimation in Monopol winter wheat, as compared with Katepwa spring wheat, is associated with the increased capacity of Monopol for sucrose utilisation through the biosynthesis of fructans in the leaves and subsequent export to the crowns. In contrast, the differential adjustment of CO₂‐saturated rates of CO₂ assimilation upon high light acclimation of Monopol appears to be associated with both increased fructan and starch accumulation in the crowns.     

Protoplast responses in the epidermis of Allium cepa induced by penetration by Botrytis allii

Penetration of Allium cepa epidermal cells (white, yellow, and red varieties) by Botrytis allii induced a response by host protoplasts in normal tissue which was not observed when penetrations were made in protoplast-free host cell walls. Callose and auto-fluorescing substances (possibly phenolic compounds) were located at the penetration sites only in normal host cells containing protoplasts. Lignin tests were negative. Halos were clearly visible in both types of tissue. Autofluorescence was observed at penetration sites in normal cells of all cultivars but general wall background autofluorescence was not observed in white onions. Autofluorescence was generally yellow green and when treated with ammonium hydroxide became green. Treatment with sodium hydroxide abolished autofluorescence. No attempt was made to isolate the autofluorescing material.     

Loss of actin cytoskeletal function and EDS1 activity,in combination,severely compromises non-host resistance in Arabidopsis against wheat powdery mildew

Plant immunity against the majority of the microbial pathogens is conveyed by a phenomenon known as non-host resistance (NHR). This defence mechanism affords durable protection to plant species against given species of phytopathogens. We investigated the genetic basis of NHR in Arabidopsis against the wheat powdery mildew fungus Blumeria graminis f. sp. tritici (Bgt). Both primary and appressorial germ tubes were produced from individual Bgt conidia on the surface of the Arabidopsis leaves. Attempted infection occasionally resulted in successful penetration, which led to the development of an abnormal unilateral haustorium. Inoculation of a series of Arabidopsis defence-related mutants with Bgt resulted in the attenuation of reactive oxygen intermediate (ROI) production and salicylic acid (SA)-dependent defence gene expression in eds1, pad4 and nahG plants, which are known to be defective in some aspects of host resistance. Furthermore, Bgt often developed bilateral haustoria in the mutant Arabidopsis lines that closely resembled those formed in wheat. A similar decrease in NHR was observed following treatment of the wild-type Arabidopsis plants with cytochalasin E, an inhibitor of actin microfilament polymerisation. In eds1 mutants, inhibition of actin polymerisation severely compromised NHR in Arabidopsis against Bgt. This permitted completion of the Bgt infection cycle on these plants. Therefore, actin cytoskeletal function and EDS1 activity, in combination, are major contributors to NHR in Arabidopsis against wheat powdery mildew.     

水稻受稻瘟菌侵染后发病初期的细胞学反应

采用致病性不同的3个稻瘟菌株接种水稻IR64品种。结果显示：在抗性反应中,病菌接种后水稻细胞中形成原生质颗粒,逐渐向病菌侵入部位聚集;原生质浓缩及沉积的形成、细胞坏死与病菌侵染菌丝的受抑制在时间和空间上是一致的。在中度抗性反应中,原生质颗粒化的时间延迟。在感病反应中,尚未观察到水稻细胞质浓缩现象。病菌侵染后寄主细胞在蓝光激发下的自发荧光表明,在寄主细胞中有多酚类物质的产生和积累。抗性反应中稻瘟菌接种后多酚类物质产生早,荧光范围大而强;中度抗性反应中,多酚类物质产生迟,荧光范围小而弱;而在感病反应中,难以观察到寄主细胞的自发荧光。胼胝质、磷脂酶Dγ(phospholipase Dγ,PLDγ)产生和积累的趋势与多酚类积累的情况大致是相似的。不同互作类型寄主细胞中多酚类物质、胼胝质和PLDγ产生和积累的差异表明,这些物质在水稻的抗病性中起了重要作用。     

诱导处理小麦对蚜虫生长发育的影响及小麦特异基因的表达

分析了麦二叉蚜在小麦98-10-30上不同诱导处理后发育时间、成虫重量和平均相对生长率,以及不同诱导处理下小麦98-10-30特异基因的表达.结果表明,用针刺、蚜虫取食和BTH（benzo(1,2,3)thiadiazole-7-carbothioic acid S-methyl ester）处理98-10-30后,蚜虫发育成成虫的时间缩短,成虫体重下降,平均相对生长率在各处理间无显著差异.对处理后小麦体内特异基因的表达研究表明,LOX、AOS、PDF1.2、PAL1、PR-1和BGL2基因在mRNA的表达量或质上存在差异.PDF1.2基因在蚜虫取食后mRNA的量显著增加,但BTH处理却不诱导PDF1.2基因的表达,同时,BGL2基因在蚜虫取食和BTH处理后,mRNA的量增加,但对照和针刺处理后BGL2基因不表达.不同诱导处理表明,98-10-30诱导的抗性对麦二叉蚜的生长发育有一定影响,蚜虫取食诱导的防卫反应与机械损伤和BTH处理有相似或相同之处,但存在明显差异.这说明蚜虫诱导的抗性是一种特异反应,与受伤反应和抗病反应有重叠交叉之处,但又不同于二者诱导产生的抗性反应.     

Photosystem II Excitation Pressure and Development of Resistance to Photoinhibition (II. Adjustment of Photosynthetic Capacity in Winter Wheat and Winter Rye)

Winter wheat (Triticum aestivum L. cv Monopol), spring wheat (Triticum aestivum L. cv Katepwa), and winter rye (Secale cereale L. cv Musketeer) grown at 5[deg]C and moderate irradiance (250 [mu]mol m-2 s-1) (5/250) exhibit an increased tolerance to photoinhibition at low temperature in comparison to plants grown at 20[deg]C and 250 [mu]mol m-2 s-1 (20/250). However, 5/250 plants exhibited a higher photosystem II (PSII) excitation pressure (0.32-0.63) than 20/250 plants (0.18-0.21), measured as 1 - qP, the coefficient of photochemical quenching. Plants grown at 20[deg]C and a high irradiance (800 [mu]mol m-2 s-1) (20/800) also exhibited a high PSII excitation pressure (0.32-0.48). Similarly, plants grown at 20/800 exhibited a comparable tolerance to photoinhibition relative to plants grown at 5/250. In contrast to a recent report for Chlorella vulgaris (D.P. Maxwell, S. Falk, N.P.A. Huner [1995] Plant Physiol 107: 687-694), this tolerance to photoinhibition occurs in winter rye with minimal adjustment to polypeptides of the PSII light-harvesting complex, chlorophyll a/b ratios, or xanthophyll cycle carotenoids. However, Monopol winter wheat exhibited a 2.5-fold stimulation of sucrosephosphate synthase activity upon growth at 5/250, in comparison to Katepwa spring wheat. We demonstrate that low-temperature-induced tolerance to photoinhibition is not a low-temperature-growth effect per se but, instead, reflects increased photosynthetic capacity in response to elevated PSII excitation pressure, which may be modulated by either temperature or irradiance.     

Benzothiadiazole (BTH) Induces Cell-Death Independent Resistance in Phaseolus vulgaris against Uromyces appendiculatus

Benzo‐(1,2,3)‐thiadiazole‐7‐carbothioic acid S‐methyl ester (BTH), trade name Bion^®, was used to induce resistance in bean cultivars Borlotto Nano Lingua di Fuoco (BLF), Borlotto Taylor, Cannellino, Cannellino Montalbano, Saxa and Top Crop, against rust caused by Uromyces appendiculatus. A single 0.3 mm BTH spray 7 days before inoculation was sufficient to fully control the disease in all the examined cultivars. As regards the more susceptible BLF, either a single treatment 14 days before inoculation, or three applications on the third, fifth and seventh day before inoculation, were equally effective to prevent infection. Histochemical and cytochemical investigations showed that BTH causes hydrogen peroxide (H₂O₂) accumulation in the treated tissues. H₂O₂ deposits were localized in situ for the first time in the apoplast of the leaf epidermis. No cell death was detected at BTH concentrations below the phytotoxicity threshold, suggesting that acquired resistance against bean rust is mainly related to the enhanced activity of anionic peroxidases, promoted by H₂O₂ accumulation, thereby leading to cell wall strengthening. This hypothesis is also supported by the long induction phase required to establish complete resistance.     

Hypersensitive lignification response as the mechanism of non-host resistance of wheat against oat crown rust

Seven-day-old seedlings of the near-isogenic wheat ( Triticum aestivum L.) lines Prelude and Prelude-Sr5, susceptible and resistant to wheat stem rust, respectively, were inoculated with uredospores of the oat crown rust fungus Puccinia coronata Cda. f. sp. avenae Fraser & Led. Fluorescence microscopy revealed that the majority of colonies developed intercellular infection structures including haustorial mother cells and haustoria after penetration of wheat mesophyll cells. All penetrated cells became necrotic, and exhibited bright yellow autofluorescence. This autofluorescence was not extractable with alkali, and fluorescent cells stained positively with phloroglucinol/HCI, suggesting that hypersensitive cell death was correlated with cellular lignification. Accordingly, the lignin biosynthetic enzymes phenylalanine ammonia-lyase (EC4.3.1.5). 4-coumarate:CoA ligase (EC6.2.1.12), cinnamyl-alcohol dehydrogenase (EC1.1.1.149), and peroxidases (EC1.11.1.7) increased in activity during the expression of resistance. The induced pattern of peroxidase iso/ymes closely resembled that observed for highly incompatible wheat/wheat stem rust interactions. Furthermore, an elieitor was extracted from oat crown rust germlings. which induces lignification when injected into the intercellular space of wheat leaves. This elieitor appears to be functionally similar to that isolated from wheat stem rust germlings. The results suggest that the non-host resistance of wheat to the xenopara-site oat crown rust closely resembles the race/cullivar-speeific resistant mechanism of highly resistant wheat varieties to wheat stem rust.     

Genetic analysis and molecular mapping of a new powdery mildew resistant gene Pm46 in common wheat

Powdery mildew (PM), caused by Blumeria graminis f. sp. tritici (Bgt), has become a serious disease and caused severe yield losses in the wheat production worldwide. Resistance gene(s) in wheat cultivars can be quickly overcome by newly evolved pathogen races when these genes are employed for long time or in a large area. It is urgent to search for new sources of resistance to be used in wheat breeding. Tabasco is a German resistant cultivar and a new source of resistance gene(s) to PM. An F(2) population was developed from a cross between Tabasco and a Chinese susceptible cultivar Ningnuo 1. Infection types in 472 F(2) plants and 436 F(2-3) families were evaluated by inoculating plants with isolate Bgt19. Results showed that a single dominant gene, designed Pm46, controlled powdery mildew resistance in Tabasco. This gene was located to the short arm of chromosome 5D (5DS) and flanked by simple sequence repeat markers Xgwm205 and Xcfd81 at 18.9?cM apart. Because another resistance gene Pm2 was also located on 5DS, 15 Bgt isolates were used to inoculate Tabasco and Ulka/8*Cc (Pm2 carrier). The results showed that Tabasco was highly resistant to all of the 15 isolates tested, while Ulka/8*Cc was susceptible to 4 of the isolates, suggesting that Tabasco may carry resistant gene(s) different from Pm2 gene in Ulka/8*Cc. To test the allelism between Pm46 and Pm2, an F(2) population between Tabasco and Ulka/8*Cc was developed. Isolate Bgt2, avirulent to both parents, was used to evaluate the F(2) population and two susceptible plants were identified from 536 progenies with F(2) plants. This result indicated that Pm46 is not allelic to Pm2. Therefore, Pm46 is a new gene for PM resistance identified in this study.     

用半定量RT-PCR方法分析小麦TaMlo-A1c基因的表达

以小麦稳定表达的肌动蛋白基因(Actin)作为对照,利用半定量反转录聚合酶链式反应(Semi-QRT-PCR)技术,对与抗白粉病有关的小麦(TriticumaestivumL.)TaMlo-A1c基因的表达进行了研究。结果发现:TaMlo-A1c基因在小麦的叶、茎、根中均表达,穗中不表达,在白粉菌(Blumeriagraminis(DC.)E.O.Speerf.sp.triticiEm.Marchal,Bgt)诱导不同时间后小麦叶片中的表达稍微有所增强。研究表明,用半定量RT-PCR技术研究小麦基因表达,具有特异性高、操作简便和可靠性强的优点。     

Independent genetic mechanisms mediate turgor generation and penetration peg formation during plant infection in the rice blast fungus

The first barrier to infection encountered by foliar pathogens is the host cuticle. To traverse this obstacle, many fungi produce specialized infection cells called appressoria. MST12 is essential for appressorium-mediated penetration and infectious growth by the rice pathogen Magnaporthe grisea. In this study, we have characterized in detail the penetration defects of an mst12 deletion mutant. Appressoria formed by the mst12 mutant developed normal turgor pressure and ultrastructure but failed to form penetration pegs either on cellophane membranes or on plant epidermal cells. Deletion and site-directed mutagenesis analyses indicated that both the homeodomain and zinc finger domains, but not the middle region, of MST12 are essential for appressorial penetration and plant infection. The mst12 mutant appeared to be defective in microtubule reorganization associated with penetration peg formation. In mature appressoria, the mutant lacked vertical microtubules observed in the wild type. The mst12 mutant also failed to elicit localized host defence responses, including papilla formation and autofluorescence. Our data indicate that generation of appressorium turgor pressure and formation of the penetration peg are two independent processes. MST12 may play important roles in regulating penetration peg formation and directing the physical forces exerted by the appressorium turgor in mature appressoria.     

Defense Reactions in Host and Nonhost Plants Against the Soybean Rust Fungus (Phakopsora pachyrhizi Syd.)

Growth and development of two races of the soybean rust fungus (Phakopsora pachyrhizi Syd.) were compared on host and nonhost plants. Both groups had several lines of defense, each of which could stop a part of attacking uredospores. Germ tubes and appressoria were produced equally well on hosts and nonhosts. A reduced formation of penetration hyphae contributed to the resistance of nonhosts and resistant host genotypes. In the epidermal cells of wheat and barley leaves, lower frequencies of penetration hyphae coincided with the production of papillae-like structures which were not penetrated. The last line of defense of all nonhosts was localized in the epidermal cell where the growth of the penetration hyphae was arrested definitively. The colony development in these species was suppressed completely. In highly resistant host genotypes the last defense reaction occurred later as a hypersensitive cell collapse which interrupted the growth of the rust colony.     

An aluminum-activated citrate transporter in barley

Soluble ionic aluminum (Al) inhibits root growth and reduces crop production on acid soils. Al-resistant cultivars of barley (Hordeum vulgare L.) detoxify Al by secreting citrate from the roots, but the responsible gene has not been identified yet. Here, we identified a gene (HvAACT1) responsible for the Al-activated citrate secretion by fine mapping combined with microarray analysis, using an Al-resistant cultivar, Murasakimochi, and an Al-sensitive cultivar, Morex. This gene belongs to the multidrug and toxic compound extrusion (MATE) family and was constitutively expressed mainly in the roots of the Al-resistant barley cultivar. Heterologous expression of HvAACT1 in Xenopus oocytes showed efflux activity for (14)C-labeled citrate, but not for malate. Two-electrode voltage clamp analysis also showed transport activity of citrate in the HvAACT1-expressing oocytes in the presence of Al. Overexpression of this gene in tobacco enhanced citrate secretion and Al resistance compared with the wild-type plants. Transiently expressed green fluorescent protein-tagged HvAACT1 was localized at the plasma membrane of the onion epidermal cells, and immunostaining showed that HvAACT1 was localized in the epidermal cells of the barley root tips. A good correlation was found between the expression of HvAACT1 and citrate secretion in 10 barley cultivars differing in Al resistance. Taken together, our results demonstrate that HvAACT1 is an Al-activated citrate transporter responsible for Al resistance in barley.