Early molecular signatures of responses of wheat to Zymoseptoria tritici in compatible and incompatible interactions

Zymoseptoria tritici, the causal agent of septoria tritici blotch, a serious foliar disease of wheat, is a necrotrophic pathogen that undergoes a long latent period. Emergence of insensitivity to fungicides, and pesticide reduction policies, mean there is a pressing need to understand septoria and control it through greater varietal resistance. Stb6 and Stb15, the most common qualitative resistance genes in modern wheat cultivars, determine specific resistance to avirulent fungal genotypes following a gene‐for‐gene relationship. This study investigated compatible and incompatible interactions of wheat with Z. tritici using eight combinations of cultivars and isolates, with the aim of identifying molecular responses that could be used as markers for disease resistance during the early, symptomless phase of colonization. The accumulation of TaMPK3 was estimated using western blotting, and the expression of genes implicated in gene‐for‐gene interactions of plants with a wide range of other pathogens was measured by qRT‐PCR during the presymptomatic stages of infection. Production of TaMPK3 and expression of most of the genes responded to inoculation with Z. tritici but varied considerably between experimental replicates. However, there was no significant difference between compatible and incompatible interactions in any of the responses tested. These results demonstrate that the molecular biology of the gene‐for‐gene interaction between wheat and Zymoseptoria is unlike that in many other plant diseases, indicate that environmental conditions may strongly influence early responses of wheat to infection by Z. tritici, and emphasize the importance of including both compatible and incompatible interactions when investigating the biology of this complex pathosystem.     

Differential abundance of proteins in response to Beet necrotic yellow vein virus during compatible and incompatible interactions in sugar beet containing Rz1 or Rz2

Rhizomania, caused by Beet necrotic yellow vein virus (BNYVV), is an important disease affecting sugar beet. Control is achieved through planting of resistant varieties; however, following the introduction of Rz1, new pathotypes that overcome resistance have appeared. To understand how BNYVV overcomes resistance, we examined quantitative protein differences during compatible and incompatible interactions when sugar beet is infected with either a traditional A-type strain or with an Rz1 resistance breaking strain. Proteomic data suggest distinct biochemical pathways are induced during compatible and incompatible sugar beet interactions with BNYVV. Pathways including the detoxification of reactive oxygen species, UB/proteasome, and photosynthesis should be studied in more depth to characterize roles in symptom development.     

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.     

Quantitative histological studies of compatible and incompatible interactions between accessions of Macroptilium atropurpureum and Uromyces appendiculatus

Quantitative histological studies of pre-penetration, penetration and post-penetration events were made for one compatible interaction and five interactions showing varying degrees of incompatibility between Uromyces appendiculatus and Macroptilium atropurpureum. Four histologically different resistant responses to infection were observed, CPI 91348 exhibited a response, characterized by significantly fewer pustules per unit area and a longer generation time than the susceptible cultivar Siratro. Significantly fewer urediniospores produced appressoria on 91348 than on Siratro. Resistance in CPI 84997 was associated with the development of large areas of necrotic tissue; 99.8% of colonies showed an indeterminant hypersensitive response. Colony areas for the above three host genotypes were not significantly different at 144 h after inoculation.
The remaining lines developed varying degrees of chlorosis only. Colonies in CQ 1382 and (CPI 92643 grew only slowly, but were still growing 144 h after inoculation and, as with CPI 84997, this growth continued after necrotic tissue appeared (indeterminant hypersensitivity), In CQ 1398, colony growth ceased by 96 144 h after inoculation, coinciding with the appearance of necrotic tissue (determinant hypersensitivity).     

Overexpression of phytoglobin in barley alters both compatible and incompatible interactions with the mildew pathogen Blumeria graminis

This study showed how barley plants can be shifted in their response to isolates of the mildew pathogen Blumeria graminis with different host adaptation by overexpression of the barley phytoglobin gene HvHb1. At early infection stages, plants overexpressing phytoglobin (GPHb1) showed less papilla formation and more hypersensitive response against both virulent and avirulent pathogen isolates compared to the wildtype (WT) plants. The shift was most pronounced in a wheat-adapted isolate (B. graminis f. sp. tritici). At later infection stages, GPHb1 plants infected with a virulent pathogen isolate (A6) showed less leaf chlorosis compared to the WT plants, indicating delayed senescence. The chlorophyll level was significantly higher in A6-infected GPHb1 plants 9 days after inoculation (dai) and the senescence indicators sphingosine-1-phosphate:ceramide ratio and phytol content confirmed delayed senescence. At 14 dai the percentage of fungal DNA was significantly higher on the GPHb1 plants than on WT plants, probably as a result of the delayed senescence. The results show that overexpression of phytoglobin (previously known as plant haemoglobin) can be an important tool to understand disease-related stress effects in plants of agronomic importance and for understanding basic resistance mechanisms. Studying this process in more detail may provide insights into how to alleviate stress-related senescence in plants.     

大麦种子萌发期对渗透胁迫的响应及 抗旱性鉴定指标的筛选

选用4份大麦材料,采用0%、5%、10%、15%、20%(W/V)PEG8000溶液模拟干旱胁迫处理,研究大麦种子萌发期发芽势、发芽率、根冠比、物质转运速率等形态生长发育指标对渗透胁迫的响应,进行抗旱性评价,筛选抗旱性鉴定指标。结果表明,随着渗透胁迫程度的增大,对大麦种子萌发产生显著的抑制作用,各指标对渗透胁迫的响应并不一致;20%PEG8000是较适宜的抗旱性鉴定浓度;初步推断胚根长、胚芽长、胚芽干重、物质转运速率可以作为大麦种子萌发期快速、有效的抗旱性鉴定指标;参试品种抗旱性强弱依次为Z027S078T、Z182U038V、Z037P017Q-1、Z039P046Q。     

不同大豆品种在萌发期对干旱胁迫的生理响应及抗旱性评价

对3个大豆品种(黄豆、晋豆19及郑9007)采用不同浓度PEG-6000(0、12%、14%、16%、18%和20%)模拟干旱胁迫处理,测定种子萌发及抗氧化酶活性等指标。结果表明,不同浓度PEG处理均降低了3个大豆品种的相对发芽势、相对发芽率及相对胚根长,18%PEG处理为合适的干旱胁迫条件。在18%PEG处理下,3个大豆品种胚根的超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性、可溶性糖、可溶性蛋白、脯氨酸及丙二醛(MDA)含量均发生了显著变化。其中,黄豆胚根SOD活性、POD活性、CAT活性、可溶性糖、可溶性蛋白、脯氨酸含量分别为对照的5.41、2.13、2.64、2.84、2.13倍和10.98倍,均高于晋豆19和郑9007,而MDA含量上升为对照的1.59倍,低于晋豆19和郑9007。通过隶属函数法综合评价发现,3个大豆品种的抗旱性强弱依次为黄豆郑9007晋豆19,说明黄豆的抗旱能力高于晋豆19和郑9007。     

不同品种藜麦幼苗对干旱胁迫的生理响应及耐旱性评价

为研究干旱胁迫对不同品种藜麦幼苗生长和生理特性的影响并评价不同品种耐旱性,本试验以4个陇藜系列藜麦品种（即‘陇藜1号’、‘陇藜2号’、‘陇藜3号’和‘陇藜4号’）为材料,设置正常灌水（CK）、轻度水分胁迫（LD,土壤含水量为田间持水量的60%~50%）、中度水分胁迫（MD,土壤含水量为田间持水量的40%~30%）和重度水分胁迫（SD,土壤含水量为田间持水量的20%~10%）4个处理,通过测定幼苗生物量指标和生理生化指标,运用数学分析方法明确测定指标与干旱胁迫的关系,并进行耐旱性评价。结果表明：SD处理各品种幼苗株高分别较CK降低了30.64%、28.36%、32.76%和37.88%,根长分别较CK增加了37.54%、81.46%、43.33%和74.85%,根冠比较CK分别升高了42.86%、33.33%、40.00%和40.00%;‘陇藜1号’ 幼苗叶片叶绿素含量分别较CK处理升高了29.40%,其余3个品种分别较CK 处理降低了33.22%、36.20%和15.99%;各供试品种幼苗叶片可溶性糖含量较CK处理分别升高了0.51%、1.67%、1.19%和2.07%;脯氨酸含量较CK处理分别升高了116.84%、268.07%、798.01%和208.70%;‘陇藜3号’幼苗叶片SOD活性较CK处理升高了6.67%;各供试品种幼苗叶片POD活性较正常处理分别升高了163.96%、62.83%、73.27%和169.50%;幼苗叶片CAT活性较CK处理分别升高了16.87%、11.44%、10.79%和4.30%。说明干旱胁迫下,藜麦幼苗通过增加渗透调节物质含量、增强抗氧化酶活性、清除体内活性氧物质,从而使幼苗适应干旱环境;但当干旱胁迫严重到一定程度时,抗氧化酶活性减弱,植株体内ROS积累,叶片叶绿素合成和干物质积累受到抑制,表现为植株生长受到影响。通过隶属函数法分析可知,供试材料抗旱性表现为‘陇藜1号’=‘陇藜3号’>‘陇藜2号’>‘陇藜4号’。     

水稻白叶枯病成株抗性与过氧化氢含量及几种酶活性变化的关系

 分析了水稻白叶枯病成株抗性与过氧化氢(H₂O₂)含量及过氧化物酶(POD)、多酚氧化酶(PPO)、超氧化物歧化酶(SOD)和过氧化氢酶(CAT)活性变化的关系。试验结果表明:接种白叶枯病菌T7133后,苗期与成株期植株体内H₂O₂含量上升以及POD、PPO和SOD活性增加。与苗期叶片相比,成株期叶片H₂O₂含量高,且PPO和SOD的酶活性增强,而POD的酶活性则降低。苗期和成株期CAT的酶活性均低于对照,成株期比苗期CAT活性更低。这些结果表明,H₂O₂、PPO、SOD和CAT可能与水稻白叶枯病成株抗性之间存在一定的关联,而POD则没有直接关系。     

硬枝碱蓬幼苗对盐旱胁迫的生理响应

以硬枝碱蓬幼苗为材料,通过NaCl和PEG6000设置不同程度的盐旱处理,对硬枝碱蓬苗期抗性生理指标进行测定,研究其的抗盐性和抗旱性。结果表明:1)与对照(0)相比,在NaCl为100~500 mmol/L范围内,随着NaCl浓度升高,硬枝碱蓬幼苗的相对电导率和丙二醛(MDA)含量无显著变化,脯氨酸含量、可溶性糖含量、可溶性蛋白含量、过氧化氢酶(CAT)活性、过氧化物酶(POD)活性呈增加的趋势;2)与对照(0)相比,在渗透势为-0.7~-0.1 MPa范围内,硬枝碱蓬幼苗的相对电导率和丙二醛含量在高渗透时无显著变化,但在低渗透势时显著增加,脯氨酸含量、可溶性糖含量、可溶性蛋白含量、CAT活性随着渗透的下降呈增加趋势,POD活性随着渗透的下降呈先下降后增加的趋势;3)综合盐旱处理下硬枝碱蓬幼苗的生理指标变化,硬枝碱蓬幼苗表现出较好的抗盐性和抗旱性,但低渗透势(-0.7 MPa)会对其幼苗造成一定程度伤害。     

干旱胁迫下五种荒漠灌木苗期的生理响应和抗旱评价

采用盆栽断水法模拟土壤干旱胁迫,研究了五种典型荒漠灌木-小叶锦鸡儿、荒漠锦鸡儿、沙冬青、花棒和霸王的生理适应特性.结果表明:干旱胁迫下,五种灌木苗期叶片保水能力和叶绿素a/b降低,可溶性糖、脯氨酸和MDA含量增加.各灌木SOD、POD和CAT活性变化不尽相同,但都相互协调作用维持在一个平衡态进行,且与MDA含量成正相关...     

Exploring host-pathogen combinations for compatible and incompatible reactions in grapevine downy mildew

Rhizoctonia leguminicola, the traditional name for the causal agent of blackpatch of red clover (Trifolium pratense) and other legumes, produces alkaloids, one of which causes livestock to salivate excessively. Fungal presence is generally confirmed by microscopy, disappearance of symptoms in livestock after removal of suspect forage, and chromatography of the alkaloid slaframine, in legume tissue. Use of the polymerase chain reaction (PCR) to amplify a pathogen-specific DNA fragment would complement the other methods of pathogen identification. Primers were designed to the R. leguminicola ITS region, sequence provided by another laboratory. Two separate primer pairs each amplified a different fragment–one ~250 bp long (expected length 249 bp), and the other 300 to 400 bp long (expected length 368 bp)–in DNA extracted from cultures of R. leguminicola. Under the experimental conditions, the primers to the larger fragment amplified a stronger band, and a minimum of 0.1 ng DNA per reaction was needed to produce a detectable band. With the primers to the 368-bp fragment, a band 300 to 400 bp long was also amplified in DNA extracted from red clover (cultivar Kenland) inoculated with R. leguminicola and harvested 70 h post inoculation. No amplification with this primer set occurred in DNA extracted from mock-inoculated red clover plants, supporting the likelihood that the primers amplified R. leguminicola DNA extracted from inoculated red clover. This primer set did not amplify DNA extracted from a red clover isolate of the legume pathogen Stemphylium sarcinaeforme, or DNA extracted from two isolates of the legume pathogen Colletotrichum trifolii, indicating specificity for R. leguminicola DNA. Lack of amplification of alfalfa DNA indicated that the R. leguminicola primers will be useful for testing for the presence of blackpatch in alfalfa.     

Microscopic detection of reactive oxygen species generation in the compatible and incompatible interactions of Alternaria alternata Japanese pear pathotype and host plants

 Reactive oxygen species (ROS) generation was examined in the interaction of Alternaria alternata Japanese pear pathotype and host plants using three methods: nitro blue tetrazolium (NBT) method for microscopic detection of O₂ ⁻, diaminobenzidine (DAB) methods for microscopic detection of H₂O₂, and cerium chloride methods for ultrastructural detection of H₂O₂. ROS generation was detected by NBT and DAB methods at appressoria on leaves of susceptible cultivars and heat-shocked leaves of resistant cultivars but not in leaves of resistant cultivars. Ultrastructural detection by the cerium chloride method identified ROS generation at cell walls of appressoria and penetration pegs in susceptible, resistant leaves and heat-shocked leaves. These differences in the ultrastructural and microscopic data in resistant areas were due to the restriction of ROS generation in limited areas, the side facing the plant surface, of appressoria and penetration pegs. Therefore, ROS generation was apparently induced regardless of the resistance or susceptibility of the cultivar with the difference being in the volumes generated. After evaluating the pathological role of ROS generation in fungal structures, such generation was found to be associated with early penetration of cell walls in pear plants. Additionally, ROS generation in plants was also found in degrading pectin layers near infected hyphae and in plasma membrane modification sites in susceptible leaves but not in resistant leaves. ROS generation in susceptible leaves might be accompanied with plasma membrane damage, although the role of ROS generation in the pectin layers is not clear. ROS generation in both fungal and plant cells during their interaction was likely associated with the expression of susceptibility. Received: June 3, 2002 / Accepted: July 31, 2002     

ROS and NO production in compatible and incompatible tomato-<Emphasis Type="Italic">Meloidogyne incognita</Emphasis> interactions

Nitric oxide (NO) has been shown to be an essential regulatory molecule in plant response to pathogen infection in synergy with reactive oxygen species (ROS). At the present, nothing is known about the role of NO in disease resistance to nematode infection. We used a resistant tomato cultivar with different sensitivity to avirulent and virulent populations of the root-knot nematode Meloidogyne incognita to investigate the key components involved in oxidative and nitrosative metabolism. We analyzed the superoxide radical production, hydrogen peroxide content, and nitric oxide synthase (NOS)-like and nitrate reductase activities, as potential sources of NO. A rapid NO accumulation and ROS production were found at 12 h after infection in compatible and incompatible tomato-nematode interactions, whereas the amount of NO and ROS gave different results 24 and 48 h after infection amongst compatible and incompatible interactions. NOS-like arginine-dependent enzyme rather than nitrate reductase was the main source of NO production, and NOS-like activity increased substantially in the incompatible interaction. We can envisage a functional overlap of both NO and ROS in tomato defence response to nematode invasion, NO and H₂O₂ cooperating in triggering hypersensitive cell death. Therefore, NO and ROS are key molecules which may help to orchestrate events following nematode challenge, and which may influence the host cellular metabolism.     

Genetic resistance to bacterial blight disease in Persian walnut

Bacterial blight disease of Persian walnut (Juglans regia, L.), caused by Xanthomonas arboricola pv. juglandis (Xaj), leads to significant nut losses in northern, central and western areas of Iran. To identify the natural sources of resistance to disease in the endemic walnut genotypes of Iran, sixteen walnut genotypes, collected from different areas of Hamedan province, were inoculated with Xaj in a randomized complete block design with five replicates for each genotype. Two-year old genotypes were gently sprayed with a suspension of bacteria adjusted to approximately 2 × 10⁹ cfu ml⁻¹ of distilled water in May. Infected leaves were rated for disease 28 and 42 days after inoculation, using a 0 to 5 severity scale, based on the number, size and distribution of lesions on the leaves. Data analyses showed that there were variations among genotypes in response to pathogen. Upon inoculation by bacterial suspension genotype 94 showed the highest resistance to both disease incidence and its progress after 4–6 weeks of infection. Genotype 65 showed high susceptibility to disease and genotype 69 showed high susceptibilities both to disease incidence and its progress after 4–6 weeks of infection.     

白茎盐生草种子萌发特性及其对盐旱胁迫的响应

以白茎盐生草种子为材料,研究不同光照、温度、NaCl和PEG处理对白茎盐生草种子萌发特性的影响.结果表明:1)白茎盐生草萌发时对光暗条件要求不严,3种光暗条件下,白茎盐生草种子均在30℃/35℃时快速达到最大萌发率而完成萌发进程.2)当NaCl浓度为100 mmol/L时,对白茎盐生草种子萌发无显著影响向,但当NaCl浓度为200～ 500mmol/L则会显著抑制白茎盐生草种子的萌发,种子的耐盐(NaCl)阈值和极限值分别为350.94和510.32mmol/L.3)随着干旱胁迫程度的增加,完成萌发进程所用时间增长,最终萌发率显著下降.-0.2 MPa的渗透势对白茎盐生草种子最终萌发率无显著影响,渗透势为-0.4～-1.2 MPa时,种子最终萌发率总体呈显著下降的趋势,种子的耐旱(PEG)阈值和极限值分别为-1.58和-3.09 MPa.综上所述,白茎盐生草种子萌发时表现出较强的抗盐性和极强的抗旱性.