Effect of Elevated Atmospheric CO2 and Temperature Increases on the Severity of Basil Downy Mildew Caused by Peronospora belbahrii Under Phytotron Conditions

Three experimental trials have been carried out on the basil (Ocimum basilicum)–downy mildew (Peronospora belbahrii) pathosystem, under phytotron conditions, to evaluate the effect of simulated elevated atmospheric CO₂ concentrations and temperatures as well as that of their interaction. Six CO₂ and temperature combinations were tested to establish their effect on disease development. The photosynthetic efficiency (PI) and chlorophyll content index (CCI) of the basil plants were monitored throughout the trials. Average disease incidence was 43.8% under standard conditions (18–22°C and 400–450 ppm of CO₂)_, while average disease severity was 22.1%. In the same temperature regime, a doubled level of CO₂ caused a significant increase in both disease incidence and severity. When temperatures ranged between 18 and 26°C, CO₂ at 800–850 ppm increased disease incidence. At the highest temperatures tested, that is at 26–30°C, which are not favourable for downy mildew development, the increase in CO₂ had no significant effect on disease incidence. A decreasing trend of PI was observed for the PI values of the inoculated plants. This trend was particularly pronounced for high CO₂ levels at the end of the experiment. In the same way as for disease development, lower values were recorded for the inoculated plants at the end of the experiment at 18–22°C for both CO₂ concentrations and at 22–26°C for 850 ppm of CO₂. The non‐inoculated plants showed higher photosynthetic efficiency than the inoculated plants. Similar trends were also observed for the CCI, thus confirming that downy mildew incidence and severity, which in particular caused foliar damage at high CO₂ concentrations, led to a decrease in the physiological performances.     

The Effect of Downy and Powdery Mildew on Grapevine (Vitis vinifera L.) Leaf Gas Exchange

The impact of powdery (Uncinula necator) and downy mildew (Plasmopara viticola) on grapevine leaf gas exchange was analysed. Gas exchange measurements (assimilation A, transpiration E, stomatal conductance g_s, intercellular concentration of CO₂C_i) were made on three different leaf materials: (i) healthy tissue of diseased leaves, (ii) infected tissue of diseased leaves, (iii) healthy tissue of healthy leaves (control treatment). Using the same source of leaf tissue, photosynthetic pigment concentration (chlorophyll a, b) and fluorescence levels (minimal fluorescence F₀, maximal fluorescence F_m and the optimal quantum yield [F_m ? F₀]/F_m) were determined to explain the mechanism of action of the two diseases on leaf assimilation. The results indicated that powdery and downy mildew reduced the assimilation rates, not only through a reduction in green leaf area (visual lesions), but also through an influence on gas exchange of the remaining green leaf tissues, determining a ‘virtual lesion’. The ratios between virtual and visual lesions were higher in powdery mildewed leaves than in the downy mildewed leaves. The photosynthetic fluorescence level (F_v/F_m) was affected by neither of the two pathogens. The reduction in intercellular concentration of CO₂ and photosynthetic pigment may explain the lower assimilation rates in the healthy tissues of powdery and downy mildewed leaves respectively.     

Elevated atmospheric carbon dioxide and ozone alter soybean diseases at SoyFACE

Human driven changes in the Earth's atmospheric composition are likely to alter plant disease in the future. We evaluated the effects of elevated carbon dioxide (CO₂) and ozone (O₃) on three economically important soybean diseases (downy mildew, Septoria brown spot and sudden death syndrome‐SDS) under natural field conditions at the soybean free air concentration enrichment (SoyFACE) facility. Disease incidence and/or severity were quantified from 2005 to 2007 using visual surveys and digital image analysis, and changes were related to microclimatic variability and to structural and chemical changes in soybean host plants. Changes in atmospheric composition altered disease expression, but responses of the three pathosystems varied considerably. Elevated CO₂ alone or in combination with O₃ significantly reduced downy mildew disease severity (measured as area under the disease progress curve‐AUDPC) by 39–66% across the 3 years of the study. In contrast, elevated CO₂ alone or in combination with O₃ significantly increased brown spot severity in all 3 years, but the increase was small in magnitude. When brown spot severity was assessed in relation to differences in canopy height induced by the atmospheric treatments, disease severity increased under combined elevated CO₂ and O₃ treatment in only one of the 3 years. The atmospheric treatments had no effect on the incidence of SDS or brown spot throughout the study. Higher precipitation during the 2006 growing season was associated with increased AUDPC severity across all treatments by 2.7 and 1.4 times for downy mildew and brown spot, respectively, compared with drought conditions in 2005. In the 2 years with similar precipitation, the higher daily temperatures in the late spring of 2007 were associated with increased severity of downy mildew and brown spot. Elevated CO₂ and O₃ induced changes in the soybean canopy density and leaf age likely contributed to the disease expression modifications.     

Genetic dissection of a TIR‐NB‐LRR locus from the wild North American grapevine species Muscadinia rotundifolia identifies paralogous genes conferring resistance to major fungal and oomycete pathogens in cultivated grapevine

The most economically important diseases of grapevine cultivation worldwide are caused by the fungal pathogen powdery mildew (Erysiphe necator syn. Uncinula necator) and the oomycete pathogen downy mildew (Plasmopara viticola). Currently, grapegrowers rely heavily on the use of agrochemicals to minimize the potentially devastating impact of these pathogens on grape yield and quality. The wild North American grapevine species Muscadinia rotundifolia was recognized as early as 1889 to be resistant to both powdery and downy mildew. We have now mapped resistance to these two mildew pathogens in M. rotundifolia to a single locus on chromosome 12 that contains a family of seven TIR‐NB‐LRR genes. We further demonstrate that two highly homologous (86% amino acid identity) members of this gene family confer strong resistance to these unrelated pathogens following genetic transformation into susceptible Vitis vinifera winegrape cultivars. These two genes, designated r esistance to P lasmopara v iticola (MrRPV1) are the first resistance genes to be cloned from a grapevine species. Both MrRUN1 and MrRPV1 were found to confer resistance to multiple powdery and downy mildew isolates from France, North America and Australia; however, a single powdery mildew isolate collected from the south‐eastern region of North America, to which M. rotundifolia is native, was capable of breaking MrRUN1‐mediated resistance. Comparisons of gene organization and coding sequences between M. rotundifolia and the cultivated grapevine V. vinifera at the MrRUN1/MrRPV1 locus revealed a high level of synteny, suggesting that the TIR‐NB‐LRR genes at this locus share a common ancestor.     

Effects of elevated atmospheric CO2 and temperature on the management of powdery mildew of zucchini

The impact of combined environmental factors, such as temperature and CO₂, on the control of the powdery mildew of zucchini, caused by Podosphaera xanthii, and of different control measures has been studied on plants grown in phytotrons. Five experimental trials were conducted, and the powdery mildew severity of both treated and untreated zucchini plants was found to be significantly affected by the interaction between temperature (three different regimes: 16–18; 18–22; 22–26°C), CO₂ (two concentrations: 400–450 and 800–850 ppm) and the treatments. However, at the end of the trials, the efficacy of all the products was not affected by the different, tested environmental conditions. Sulphur consistently provided the highest disease control (75%–85% efficacy). Among the resistant inducers that were tested, calcium oxide was the most effective, in terms of powdery mildew control under all the conditions tested in phytotrons, reducing disease severity from 46% to 61%. Foliar applications of phosphite (14%–28% efficacy), Ampelomyces quisqualis (12%–23% efficacy) and potassium silicate (13%–24% efficacy) only slightly reduced the disease severity for all the tested temperature regimes and CO₂ concentrations, compared to the untreated control. The results obtained under our experimental conditions show that a possible increase in CO₂ concentration and temperature, which is expected for the next few years, should not influence the efficacy of the tested resistance inducers or of sulphur against powdery mildew on zucchini. Moreover, the suppressive effect of calcium oxide is in light of its possible use in greenhouses for zucchini powdery mildew control under 400–450 ppm of CO₂ and under enriched condition of 800–850 ppm of CO₂.     

An ethoxylated surfactant enhances the penetration of the sulfated laminarin through leaf cuticle and stomata,leading to increased induced resistance against grapevine downy mildew

Some β‐1,3‐glucans and particularly sulfated laminarin (PS3) are known as resistance inducers (RIs) in grapevine against the downy mildew. However, their efficacy in vineyard is still often too low, which might be caused by a limited penetration through the leaf cuticle following spray application. We used ¹⁴C‐sucrose uptake experiments with grapevine leaves in order to select a surfactant as saccharide penetration enhancer. Our results showed that although sucrose foliar uptake was low, it was strongly enhanced by Dehscofix CO125 (DE), a highly ethoxylated surfactant. Fluorescent saccharides were then produced and laser scanning microscopy was used to analyze their foliar diffusion pattern in Arabidopsis thaliana and grapevine. Interestingly, sucrose and PS3 were seemingly able to penetrate the leaf cuticle only when formulated with DE. Diffusion could preferentially occur via stomata, anticlinal cell walls and trichomes. In grapevine, PS3 penetration rate was much higher on the stomateous abaxial surface of the leaf than on the adaxial surface. Finally, using DE allowed a higher level of downy mildew control by PS3, which corroborated diffusion observations. Our results have practical consequences for the improvement of treatments with saccharidic inducers on grape. That is, formulation of such RIs plays a critical role for their cuticular diffusion and consequently their efficacy. Also, spray application should preferentially target the abaxial surface of the leaves in order to maximize their penetration.     

不同品种葡萄抗霜霉病特性与叶片POD、PPO活性关系的研究

在霜霉病盛发期,对8804、梅尔诺、品丽珠3个葡萄品种(系)叶片中的PPO和POD活性变化进行了测定.结果显示,8804的PPO和POD活性较大,并保持相当长时间的高活性值,而梅尔诺、品丽珠叶片中PPO和POD活性较小;8804的PPO酶活性变化范围高于其它2个品种,但POD酶活性变化范围却低于后者.葡萄叶片中PPO和POD活性与葡萄霜霉病抗性之间存在一定的相关性,且不同抗感品种间PPO和POD酶活性存在极显著差异.研究结果表明,8804较梅尔诺、品丽珠对霜霉病具有较强的抗性.     

Future CO2 concentrations, though not warmer temperatures, enhance wheat photosynthesis temperature responses

The temperature dependence of C3 photosynthesis is known to vary according to the growth environment. Atmospheric CO2 concentration and temperature are predicted to increase with climate change. To test whether long-term growth in elevated CO2 and temperature modifies photosynthesis temperature response, wheat (Triticum aestivum L.) was grown in ambient CO2 (370 micromol mol(-1)) and elevated CO2 (700 micromol mol(-1)) combined with ambient temperatures and 4 degrees C warmer ones, using temperature gradient chambers in the field. Flag leaf photosynthesis was measured at temperatures ranging from 20 to 35 degrees C and varying CO2 concentrations between ear emergence and anthesis. The maximum rate of carboxylation was determined in vitro in the first year of the experiment and from the photosynthesis-intercellular CO2 response in the second year. With measurement CO2 concentrations of 330 micromol mol(-1) or lower, growth temperature had no effect on flag leaf photosynthesis in plants grown in ambient CO2, while it increased photosynthesis in elevated growth CO2. However, warmer growth temperatures did not modify the response of photosynthesis to measurement temperatures from 20 to 35 degrees C. A central finding of this study was that the increase with temperature in photosynthesis and the photosynthesis temperature optimum were significantly higher in plants grown in elevated rather than ambient CO2. In association with this, growth in elevated CO2 increased the temperature response (activation energy) of the maximum rate of carboxylation. The results provide field evidence that growth under CO2 enrichment enhances the response of Rubisco activity to temperature in wheat.     

Association of RGA-SSCP markers with resistance to downy mildew and anthracnose in grapevines

Downy mildew (Plasmopara viticola) and anthracnose (Sphaceloma ampelinum) are two major diseases that severely affect most grapevine (Vitis vinifera) cultivars grown commercially in Thailand. Progress of conventional breeding programs of grapevine for improved resistance to these diseases can be speeded up by selection of molecular markers associated with resistance traits. We evaluated the association between 13 resistance gene analog (RGA)-single-strand conformation polymorphism (SSCP) markers with resistance to downy mildew and anthracnose in 71 segregating progenies of seven cross combinations between susceptible cultivars and resistant lines. F(1) hybrids from each cross were assessed for resistance to downy mildew and anthracnose (isolates Nk4-1 and Rc2-1) under laboratory conditions. Association of resistance traits with RGA-SSCP markers was evaluated using simple linear regression analysis. Three RGA-SSCP markers were found to be significantly correlated with anthracnose resistance, whereas significant correlation with downy mildew resistance was observed for only one RGA-SSCP marker. These results demonstrate the usefulness of RGA-SSCP markers. Four candidate markers with significant associations to resistance to these two major diseases of grapevine were identified. However, these putative associations between markers and resistance need to be verified with larger segregating populations before they can be used for marker-assisted selection.     

Surfactin and fengycin contribute to the protection of a Bacillus subtilis strain against grape downy mildew by both direct effect and defence stimulation

Bacillus subtilis GLB191 (hereafter GLB191) is an efficient biological control agent against the biotrophic oomycete Plasmopara viticola, the causal agent of grapevine downy mildew. In this study, we show that GLB191 supernatant is also highly active against downy mildew and that the activity results from both direct effect against the pathogen and stimulation of the plant defences (induction of defence gene expression and callose production). High-performance thin-layer chromatography analysis revealed the presence of the cyclic lipopeptides fengycin and surfactin in the supernatant. Mutants affected in the production of fengycin and/or surfactin were thus obtained and allowed us to show that both surfactin and fengycin contribute to the double activity of GLB191 supernatant against downy mildew. Altogether, this study suggests that GLB191 supernatant could be used as a new biocontrol product against grapevine downy mildew.     

供氮和增温对倍增二氧化碳浓度下荫香叶片光合作用的影响

供给0～0.6 mg N的盆栽荫香(Cinnamomum burmannii)幼树分别生长在倍增CO ₂(+CO₂,731 μmol·mol^-1)和正常空气CO ₂浓度(CO ₂,365 μmol·mol^-1)的生长箱内,昼夜温度分别为25/23 ℃和32/25 ℃,自然光照下生长30 d.以生长在CO₂和25/23 ℃下的植株为对照研究增温和氮对+CO₂叶片光合作用的影响.结果表明,在+CO₂和25/23 ℃下无氮和氮处理植株的平均光合速率(Pnsat)较+CO₂和32/25 ℃下的叶片高5.1%,温度增高降低叶片Pnsat;而Pnsat随供氮而增高.在+CO₂条件下,生长在32/25 ℃下的叶片Rubisco最大羧化速率(Vcmax)和最大电子传递速率(Jmax)较25/23 ℃下的低(P＜0.05),温度增高降低+CO₂下叶片的Vcmax和Jmax在+CO2下叶片光合呼吸速率(Rp)较低,生长温度增高提升Rp.在CO₂下生长温度从25/23 ℃增至32/25 ℃,叶片的Rubisco含量(NR)和Rubisco活化中心浓度(M)降低,而供氮能增高NR和M.供氮能减缓温度增高对倍增CO₂下荫香叶片光合作用的限制.     

Passive heat treatment of sweet basil crops suppresses Peronospora belbahrii downy mildew

Sweet basil (Ocimum basilicum) is an annual herb crop grown in polyethylene‐covered structures in Israel. It is Israel's leading herb crop, grown in warm regions of the country. Downy mildew (caused by Peronospora belbahrii) is a severe disease in Israel and in many other crop‐growing regions worldwide. Experiments were carried out to identify potential climate‐management techniques for suppression of this disease on basil in non‐heated greenhouses. Disease severity was evaluated under commercial‐like conditions in three experiments, with 8–10 walk‐in tunnels at each location. Pathogen inoculum was introduced into all walk‐in tunnels. Regression analysis was performed between the disease values and air temperature, relative humidity (RH) and soil temperature. Downy mildew severity was negatively related to high (>25°C) air temperature, RH in the range of 65–85% and high (>21°C) soil temperature. The increase in air temperature did not result in a significant increase in leaf temperature; canopy surface median temperatures only reached 30°C. Symptomless plants from relatively warmer tunnels (peak temperatures of 45–48°C) that were transferred to conditions that promote downy mildew (22 ± 2°C, RH > 95%) became severely diseased, showing sporulation of P. belbahrii, suggesting that infection occurred but at the high temperatures symptom expression/tissue colonisation was suppressed. Pot experiments in which aerial and subterranean plant organs were differentially heated revealed that treating the roots with a high temperature (26–31°C), similar to the soil temperatures in the warmer greenhouses, while maintaining the upper plant parts at ambient temperature (20°C), suppresses canopy downy mildew. The effect lasted for 1–2 weeks after the plants were removed from the heated soil treatments and maintained under optimal conditions for pathogen development. Furthermore, oospores were found in the symptomatic leaves. Oospores are minimally affected by high temperature, and therefore the high temperature presumably did not affect pathogen survival. In conclusion, the effect of high greenhouse temperature on basil downy mildew may not result from a direct negative effect of high temperature on the pathogen but from an indirect high‐temperature effect on the host, rendering it less susceptible to pathogen development.     

亚热带森林四种建群树种叶片叶肉导度对适度高温的响应

利用CO2气体交换和叶绿素荧光法,研究了适度高温(38℃)下亚热带森林4种建群树种叶片叶肉导度(gm).结果表明:阳生或早期演替树种荷木叶片gm较中后期演替的中生性或耐荫树种红椎、藜蒴和黄果厚壳桂高(P<0·05).生长在全日光下的叶片gm亦较遮荫的高(P<0·05).温度从25℃升至38℃,4个树种的平均叶肉导度Q10值为(1·59±0·27).gm对温度的响应依赖于植物种类和光环境.由于CO2在水中扩散的Q10值为1·25,gm除受温度影响外,还受到蛋白质的调控.适度高温可引起gm上升,导致叶绿体羧化部位的CO2浓度和羧化速率增高.无论是全日照还是遮荫处理,与荷木相比,适度增温可显著提高红椎、藜蒴和黄果厚壳桂叶绿体羧化速率,说明适度高温对演替中后期树种的演替进程有利.     

持续低温弱光对黄瓜叶片气体交换、叶绿素荧光猝灭和吸收光能分配的影响

持续常温弱光(25℃/18℃,l00umol m-2 s-1)、低温弱光(12℃/12℃,100 umol m-2 s-1和7℃/7℃,l00μmolm-2s-1)均导致黄瓜生长减慢或停滞、叶绿素含量、气孔导度和净光合速率、光合电子传递速率下降以及胞间CO2浓度上升.常温弱光和12℃弱光处理对光系统II的最大光化学效率Fv/Fm无显著影响,而7℃弱光处理导致Fv/Fm的可逆性下降.常温弱光和7℃、12℃弱光处理均导致了光化学反应速率的降低以及天线热耗散和反应中心过剩能量的增加.在胁迫后,12℃弱光0比7℃弱光更有利于植株光合功能的恢复.     

Loop-mediated isothermal amplification assay for the detection of Plasmopara viticola infecting grapes

Grapes downy mildew caused by obligate oomycete plant pathogen Plasmopara viticola is a devastating disease worldwide, resulting in significant yield and quality losses. A field survey was conducted in two major grapes cultivated areas of Tamil Nadu for the incidence of grapevine downy mildew. The disease incidence was 43.42%–76.69%, and the highest disease incidence of 76.69% was observed in the Theni district. Totally eight P. viticola isolates were collected from different places in Coimbatore and Theni districts. These isolates were confirmed through microscopic observation and sequencing of COX 2 gene, and the phylogenetic tree was developed to study their phylogenetic relationship among the isolates which shows 97–100% sequence similarity with other P. viticola isolates and less sequence similarity with Plasmopara species. The loop-mediated isothermal amplification (LAMP) assay was developed based on the CesA4 gene sequence of P. viticola. The assay developed was more sensitive as it detected P. viticola genomic DNA up to 20 fmg. LAMP assay specificity was proved by carrying out the assay with genomic DNA extracted from other Oomycetes and fungal plant pathogens. Finally, LAMP assay was validated by testing seventy-eight grapevine leaf samples collected from seven different locations. LAMP assay showed a positive reaction in sixty-two samples tested out of seventy-eight samples tested. Therefore, the LAMP assay described should helpful for early and specific detection of downy mildew pathogen and help in mitigating disease incidence.     

籽用美洲南瓜叶片对白粉病菌的抗逆生理响应

以籽用美洲南瓜(Cucurbita pepo L.)白粉病抗病品系F2和感病品系M3为试材,在人工气候箱内接种白粉病生理小种2US孢子悬浮液,考察在接种白粉病菌后南瓜幼苗植株与白粉病菌的互作、叶片活性氧代谢及保护酶活性的变化,探讨南瓜抵御白粉病的生理机制。结果表明：(1)与感病品系M3相比,接种白粉病菌后,抗病品系F2叶片上病原菌发育缓慢,较难侵染叶片。(2)抗病品系F2在感病初期叶片H₂O₂、O₂^-·含量迅速升高后逐渐下降,而感病品系在感病初期H₂O₂、O₂^-·含量上升缓慢,在达最大值后始终保持较高水平,且感病品系叶片MDA含量始终高于抗病品系;组织化学染色分析发现,抗病品系叶片着色比感病品系快,之后着色面积有所减少并趋于较低水平。(3)抗病品系F2和感病品系M3叶片抗氧化酶CAT、SOD、POD活性及PAL、PPO活性在接种白粉病菌后均显著增加,但抗病品系的活性及其增幅均高于感病品系。研究发现,籽用美洲南瓜抗病品系叶片上白粉病菌发育缓慢,较难受到侵染,生成菌丝体后叶片上粉状斑点较小;抗病品系在被白粉病菌侵染初期依靠活性氧的增加抵御病原菌的入侵,随着活性氧含量增加抗病品系通过迅速增加自身抗氧化酶活性来防止氧化胁迫;与感病品系相比,抗病品系在受病原菌侵染后能迅速增加PAL、PPO活性以抵御病原菌侵染。