On the apple proliferation symptom display and the canopy colonization pattern of “<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma mali” in apple trees

Notwithstanding the availability of several different real time PCR protocols for “Candidatus Phytoplasma mali”, it is still unclear how informative is the estimation of the concentration of phytoplasma cells in the leaves of apple proliferation infected trees, and how the reliability of the estimations may be affected by an erratic and uneven distribution of the pathogen in the host. Here we investigated these issues systematically and showed that phytoplasma concentration varies significantly among seasons, but not between two cultivars that appeared to have different degree of susceptibility on the basis of the symptoms displayed. In fully symptomatic trees sampled at the end of the season the phytoplasmas were detectable in most leaves, but in more than half of the leaves at low concentrations. Both the pattern of colonization of the canopy and the amount of phytoplasmas varied greatly in trees that show symptom remission, although a direct relation between symptom severity and colonization could not be established. The sampling of the apple canopy for the purpose of evaluation of concentration of “Candidatus Phytoplasma mali” should take into consideration the complex pattern of colonization and seasonal variation.     

Detection of the fire blight biocontrol agent <Emphasis Type="Italic">Bacillus subtilis</Emphasis> BD170 (Biopro<Superscript>®</Superscript>) in a Swiss apple orchard

Fire blight, caused by Erwinia amylovora, is a major disease threat to apple, pear and other pome fruit worldwide. The disease is widespread in Europe and has recently become established in Switzerland. Antibiotics are the most effective controls used in North America but these are not permitted for agricultural use in most European countries. A newly registered biological control product Biopro^®, based on the antagonist Bacillus subtilis strain BD170, is being used as an alternative strategy for fire blight management. A specific molecular marker was developed for monitoring the spread of this agent on blossoms after Biopro^® spray application in a Swiss apple orchard throughout the bloom period for 2years. Direct spraying resulted in efficient primary colonisation of pistils in flowers that were open at the time of treatment. Subsequent bacterial dissemination (secondary colonisation) of flowers that were closed or at bud stage at the time of treatment was observed but was found to be dependent on the timing of treatments relative to bloom stage in the orchard. Foraging honeybees were shown to be disseminators of Biopro^®. We also report detection of the biocontrol agent in honey collected from hives where bees were exposed by placing Biopro^® at the entrance or in the hatching nest and from hives that were simply placed in sprayed orchards.     

Effect of polyploidisation on the response of apple (<Emphasis Type="Italic">Malus</Emphasis> × <Emphasis Type="Italic">domestica</Emphasis> Borkh.) to <Emphasis Type="Italic">Venturia inaequalis</Emphasis> infection

Spinach is one of the most nutritious green-leaf vegetables. In the spinach production, diseases cause a significant loss in both yield and quality. Improving disease resistance is one of the major challenges in spinach breeding. Arabidopsis nucleoporin CONSTITUTIVE EXPRESSER OF PATHOGENESIS-RELATED GENES 5 (CPR5) functions as a negative regulator of plant cell death and immunity as cpr5 mutant exhibits spontaneous cell death and heightened immunity. In addition, CPR5 play a role in trichome development as the majority of cpr5 mutant trichomes are branchless whereas wild type trichomes are often three-branched. In the spinach genome, we identified a homolog of Arabidopsis CPR5, referred to as Spinacia oleracea CPR5 (SoCPR5). To investigate the function of SoCPR5, we introduced SoCPR5 into Arabidopsis cpr5 mutant. Our data showed that both spontaneous cell death and heightened immunity were suppressed in the SoCPR5-transgenic cpr5 mutants, verifying that SoCPR5 functions as its Arabidopsis counterpart in plant cell death and immunity. SoCPR5 also fully restored wild type trichome phenotype of the cpr5 mutant. Our study therefore indicates that the function of SoCPR5 is conserved between plant species and SoCPR5 can be applied for genetic manipulation of plant immunity in spinach.     

Development of a qPCR test to detect and quantify Elsinoë piri in unsprayed and organic apple orchards and assessment of apple cultivar susceptibility to the disease

Recently, Elsinoë piri has become an important pathogen in unsprayed and organic apple orchards. The increasing demand for organic apples as well as the limited use of fungicides imposed by European legislation has turned this pathogen, which has been known for over a century in Europe, into a resurgent phytosanitary threat. It also represents a new trait to be taken into account in apple breeding programmes. As E. piri is extremely difficult to isolate and causes symptoms that can be confused with those caused by other pathogens, a quantitative PCR (qPCR) test has been developed based on the internal transcribed spacer (ITS) region of the rDNA gene. This test, which displays a low limit of detection, allows the early detection of the fungus in apple leaves. As a quantitative method, it is a promising tool for breeding purposes as it can be used on leaves to assess the level of quantitative resistance of apple cultivars to the disease. The molecular test is also useful to gain knowledge on the epidemiology of this re-emerging fungal disease. Spore trapping conducted in 2015, 2016 and 2020 using Rotorod devices and the qPCR test showed that airborne inoculum is released at the end of the growing season. Tests were also carried out to demonstrate that the pathogen was detected in buds during winter. A comparison of 18 E. piri isolates based on the sequencing of five genome regions highlighted a high genotypic diversity. All these isolates were detected with the qPCR test developed in this study.     

Translocation of apple proliferation phytoplasma via natural root grafts – a case study

Apple proliferation (AP), a phytoplasma-induced disease of apple trees, was proven to be transmitted through infected grafting material and sap-sucking insects. To date there are little firm data on disease propagation in the field via natural root grafts. This question was thus addressed in the present case study by investigating trees of a 24-year old commercial apple orchard (‘Red Chief’ on MM 111), where the existence of root connections was discovered accidentally. After having displayed specific AP symptoms, nine trees were cut down and the stubs were infiltrated or brushed with glyphosate. Herbicide injury, however, remained not only restricted to the treated stubs, but also spread to approximately 50 neighbouring trees. Surprisingly, none of the pollinators (‘Granny Smith’ on M 9) growing interjacently and alternating between herbicide-damaged main crop trees was affected. Respective to the position of the nine AP-infected and glyphosate-treated cut stumps, four sections in the orchard were defined, from which a total of 122 trees was sampled and analysed using qualitative real-time PCR for detection of AP phytoplasma. The pathogen was found in 71.4% of ‘Red Chief’ trees with severe herbicide damage and 18.8% of trees with partial herbicide damage. None of the 31 investigated pollinators was AP-infected. Our data indicate that root connections seem to play a role for the spread of AP phytoplasma at least in older orchards and between trees on vigorous rootstocks.     

Size does matter – susceptibility of apple for grey mould is affected by cell size

Apple is a seasonal product that is stored for long periods of time, during which fungal-caused decay can occur. Previous infection experiments of intact Jonagored apple with Botrytis cinerea displayed apparent differences in lesion expansion rate with respect to inoculation position on the fruit (on, above or below the equator). The goal of this study was to investigate whether these differences are consistent or not and if so, to relate them to fruit characteristics. The study involved measuring the hue angle of the intended inoculation spots prior to inoculation, and firmness and total soluble solids content of fruit from the same batch. Results showed that firmness correlated somewhat (−0.55 and −0.72 for shadow and sun side, respectively) with lesion diameter expansion rate. In a subsequent step, X-ray imaging was carried out for samples from each position. Analysis of 3D reconstructions by microcomputed tomography of these diffraction images showed that cell size was strongly correlated (0.996) to lesion diameter expansion rate. Finally, it was investigated if cell size could also be used to rank different apple cultivars for their susceptibility to B. cinerea. The result shows that there is a clear distinction between Jonagold and Golden Delicious (non-blushing variety), which have a smaller cell size, and Braeburn and Kanzi, which have a larger cell size (overall correlation of 0.87). This indicates that cell size may also be an important factor in determining susceptibility across cultivars.     

Managing apple scab (Venturia inaequalis) and powdery mildew (Podosphaera leucotricha) using Adem™

Current control of scab and mildew of apple in the UK requires the routine application of fungicides at 7 - 14-day intervals to achieve the blemish-free fruit required by the market. Such practices are generally effective, but with increased public concern about pesticides and rising costs to the grower, they are now less acceptable. The use of disease-warning systems offers scope for optimizing fungicide use by better timing of sprays. Adem^? is a PC-based system that warns of the risk of scab, mildew, Nectria fruit rot and canker and fireblight. In a 'key stage' strategy, warnings by Adem^? for scab and mildew integrated with practical pest and disease control resulted in similar or better control than a routine programme, with the additional benefit of reduced fungicide inputs and costs even in seasons exceptionally favourable for these diseases. Maximum savings in fungicide use were made by applying sprays curatively in response to scab warnings. However, this approach resulted in increased disease incidence to the crop and scab infection of fruit even in seasons unfavourable for scab attack.     

新疆额尔齐斯河流域“2005•6•1”大洪水分析

2005年6月1-2日,额尔齐斯河全流域发生了突发性融雪和降雨混合型大洪水,各支流洪水峰高、量大、来势凶猛,其中额尔齐斯河源头支流--库依尔特斯河发生有实测资料（50 a）以来最大洪水,布尔津河、哈巴河发生有实测资料（54 a）以来第2、第3位洪水,给当地造成较大经济损失。根据河流实测洪水、降水量、气温、高空和环流形势等资料,对这次洪水的成因、过程、特点进行分析。结果表明：① 在额尔齐斯河流域融雪与降雨混合型大洪水的形成过程中,前期气温和大尺度降水天气起重要作用;② 从[JP2]有实测洪水记录的50多年以来,第4次发生全流域性洪水,在全球气候变暖和随阿勒泰地区冬季降水量明显增多的形势下,灾害性洪水出现的可能性将增加;③ 由于融雪与降雨混合型洪水过程较长、峰量较大,因此,洪水挟沙能力较强,在设计蓄水工程时要引起重视;④ 对下游有较大城市或重点水利工程的山区河流,要加强防洪预警系统的建设力度,以减轻灾害性洪水造成的损失。     

Characterization of the defence response to Venturia inaequalis in ‘Honeycrisp’ apple,its ancestors,and progeny

The apple cultivar Honeycrisp exhibits genetic resistance to apple scab. The characterization of the macroscopic and microscopic responses on leaves infected by the pathogen Venturia inaequalis is described. The macroscopic resistance reactions observed in ‘Honeycrisp’, its parent ‘Keepsake’, and grandparents ‘Frostbite’ and ‘Northern Spy’ ranged from 0 (no reaction) to chlorotic flecking, stellate chlorosis, necrotic flecking, and sporulation. No hypersensitive response was observed. The resistance response occurred as early as 7 days post inoculation (dpi) at the same time that susceptible plants exhibited macroscopic signs of the disease. The resistance reactions were similar in the progeny population of ‘Honeycrisp’ × ‘Twin Bee Gala’, although they were delayed to 10–14 dpi possibly due to variable greenhouse conditions. This population segregated 3 resistant:1 susceptible, which suggests the presence of two genes in ‘Honeycrisp’ and agrees with the finding that different responses within ‘Honeycrisp’ to mixed inoculum are due to differential recognition of pathogen effectors.     

Mechanisms of induced resistance in lettuce against <Emphasis Type="Italic">Bremia lactucae</Emphasis> by DL<Emphasis Type="Bold">-</Emphasis>β<Emphasis Type="Bold">-</Emphasis>amino<Emphasis Type="Bold">-</Emphasis>butyric acid (BABA)

BABA induced local and systemic resistance in lettuce (Lactuca sativa) against the Oomycete Bremia lactucae. Structure-activity analysis showed no induced resistance by related amino-butanoic acids or β-alanine. The R-enantiomer of BABA induced resistance whereas the S-enantiomer did not, suggesting binding to a specific receptor. Other compounds known to be involved in SAR signaling, including abscisic acid, methyl-jasmonate, ethylene, sodium-salicylate and Bion® (BTH) did not induce resistance. Systemic translocation of ¹⁴C-BABA and systemic protection against downy mildew were tightly correlated. BABA did not affect spore germination, appressorium formation, or penetration of B. lactucae into the host. Epifluorescence and confocal microscopy revealed that BABA induced rapid encasement with callose of the primary infection structures of the pathogen, thus preventing it from further developing intercellular hyphae and haustoria. Invaded host cells treated with BABA did not accumulate phenolics, callose or lignin, or express HR. In contrast, cells of genetically-resistant cultivars accumulated phenolics, callose and lignin and exhibited HR within one day after inoculation. The callose synthesis inhibitor DDG did not inhibit callose encasement nor compromised the resistance induced by BABA. PR-proteins accumulated too late to be responsible for the induced resistance. DAB staining indicated that BABA induced a rapid accumulation of H₂O₂ in the penetrated epidermal host cells. Whether H₂O₂ stops the pathogen directly or via another metabolic route is not known.     

Does <Emphasis Type="Italic">Puccinia hemerocallidis</Emphasis> regularly host-alternate between <Emphasis Type="Italic">Hemerocallis</Emphasis> and <Emphasis Type="Italic">Patrinia</Emphasis> plants in Japan?

Daylily rust fungus, Puccinia hemerocallidis, was proven to host-alternate between a wild daylily, Hemerocallis fulva var. longituba, and a patrinia, Patrinia villosa. No proof was obtained for the early belief that the fungus is pathogenic to plantainlilies, Hosta species, in addition to daylilies, Hemerocallis species. The fungus seems to alternate regularly between daylilies and patrinias in Japan because most daylily species are deciduous, and a vegetatively reproducing stage of the pathogen does not seem capable of successfully overwintering free of the living host tissue.     

苄&#183;丙&#183;异WP防除直播水稻田杂草的效果

在水稻直播田用38％苄&#183;丙&#183;异WP25～50g／667m^2,不仅对稗草、千金子有特效,而且对稻田常见的鸭舌草、节节菜、异型莎草、陌上菜、牛毛毡等杂草均有很好的防除效果。38％苄&#183;丙&#183;异WP易对水稻秧苗产生不同程度白化现象,但后期会恢复,对秧苗素质及分蘖无不良影响。     

大棚菜防治"二害"措施

大棚菜的“二害”即肥分危害 ,高温危害。对大棚菜产量、产值影响很大 ,防治措施如下 :一、肥分危害的防治１．深施肥料。化肥宜深施或溶解在水中施。２．适量施肥。大棚内流失少 ,施肥量原则上低于露地蔬菜。３．施熟有机肥。施用有机肥 ,特别是人畜粪尿或栏肥要充分腐熟发酵后施用。４．适时换气。不管施用哪种肥料 ,皆不宜于傍晚进行 ,施肥后要加强通风换气 ,夜间也要保持一定的通风量 ,以防止肥料分解时产生有害气体在棚内积累 ,对蔬菜产生毒害。５．施肥距离要适宜。追施尿素等速效肥料不宜离蔬菜根部太近 ,以防烧根。作叶肥施用 ,浓度…     

Seasonal colonisation of apple trees by ‘Candidatus Phytoplasma mali’ revealed by a new quantitative TaqMan real-time PCR approach

Apple proliferation (AP), caused by ‘Candidatus Phytoplasma mali’, is an economically important disease affecting many apple-growing areas in Europe. A new TaqMan real-time PCR assay was established for absolute quantification of ‘Ca. P. mali’ by using a single-copy gene of the host plant as a reference, which is amplified with the pathogen DNA in a single-tube reaction. Normalised estimates of phytoplasma concentration are ultimately expressed as the number of phytoplasma cells per host plant cell. The assay was used to monitor the ‘Ca. P. mali’ titre over the course of two growing seasons in roots and branches of symptomatic and asymptomatic but AP-positive apple trees. All 252 root samples from symptomatic and asymptomatic trees tested positive, with an average number of 59.8 ± 5.68 (standard error) and 55.1 ± 9.83 ‘Ca. P. mali’ per host cell, respectively. From the 378 shoot samples analysed, 81% of the symptomatic and only 20% of the asymptomatic samples were AP-positive with an average number of 9.4 ± 1.04 and 0.7 ± 0.13 ‘Ca. P. mali’ per host cell, respectively. This strengthens evidence that not the pathogen occurrence alone but the presence of a certain quantity of ‘Ca. P. mali’ in the aerial tree sections is involved in symptom expression. In addition, pronounced seasonality of the phytoplasma concentration was found, not only in branches, but also for the first time in roots of symptomatic and asymptomatic apple trees. Highest phytoplasma levels in roots were detected from December to May.     

First report of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma malaysianum’ associated with <Emphasis Type="Italic">Elaeocarpus</Emphasis> yellows of <Emphasis Type="Italic">Elaeocarpus zollingeri</Emphasis>

“Elaeocarpus yellows” (ELY) is a widely reported phytoplasma disease of Elaeocarpus zollingeri trees in Japan. The phytoplasma associated with ELY (ELY phytoplasma) had not been identified at the species level because its 16S rRNA sequence had yet to be reported. Here, we report the results of a sequence analysis based on 16S rRNA and secA gene sequences, which showed that the ELY phytoplasma is related to ‘Candidatus Phytoplasma malaysianum’. To our knowledge, this is the first report showing the occurrence of ‘Ca. P. malaysianum’ outside Malaysia and the infection of E. zollingeri by the phytoplasma.     

48％柴油&#183;毒死蜱

本产品对害虫除具有胃毒和触杀作用外，还具有较强的渗透作用，同时对害虫有黏着性，用于防治十字花科蔬菜菜青虫。     

敦煌地区“6•16”暴雨的热力和动力特征

使用高空和地面定时观测资料、区域站降水资料、云图和NCEP 1°×1°再分析资料,对2011年6月15-16日敦煌暴雨的影响系统、干侵入、垂直涡度收支、视热源和视水汽汇进行了分析。结果表明：这次暴雨是中高纬度天气系统与低纬度天气系统相互作用,在敦煌附近上空形成中尺度涡旋造成的结果。在低涡生成初期,低涡南侧对流层高层有明显的干侵入发生;低涡快速发展期,干侵入达到最强;对流层中下层辐散作用项对垂直涡度正贡献最大,在对流层中层水平平流项和垂直输送项对垂直涡度正贡献起主要作用;低涡成熟期,干侵入开始减弱。对流层下层辐散作用项对垂直涡度正贡献进一步加强,对流层中层水平平流项正贡献作用减弱,垂直输送项正贡献作用增强。视热源与视水汽汇加热强度变化与低涡发生、发展的不同阶段紧密相关,视热源、视水汽汇中心与垂直上升运动中心相对应,说明大气加热与大气上升运动密切相关,水汽凝结潜热释放加速垂直上升运动,垂直输送在视热源、视水汽汇中起主要作用。     

The first detection of ‘<Emphasis Type="Italic">Candidatus</Emphasis> Phytoplasma trifolii’ in <Emphasis Type="Italic">Rhododendron</Emphasis> hybridum

Four Rhododendron hybridum plants (from cvs Moravanka and Don Juan), all exhibited symptoms of shortened axillary shoots, reduced leaves with vein clearing and yellowing, undeveloped flowers, and general stunting in a rhododendron nursery garden in southern Bohemia in 2007. Electron microscopy examination of ultra-thin sections revealed the presence of numerous polymorphic phytoplasma-like bodies in the phloem tissue of leaf midribs and petioles. The phytoplasma etiology of this disease was further confirmed by polymerase chain reaction (PCR) using universal phytoplasma primers. Restriction fragment length polymorphism (RFLP) analysis of amplification products obtained with a R16F2/R16R2 primer pair from all symptomatic plants indicated the presence of phytoplasma from the 16SrVI-A subgroup. A detailed comparison of the amplified sequences and phylogenetic analysis confirmed that the phytoplasma belonged to the subgroup 16SrVI-A (clover proliferation phytoplasma group). This is the first report of the natural occurrence of ‘Candidatus Phytoplasma trifolii’ in plants of Rhododendron hybridum.