Relationship between resistance to pine wilt disease and the migration or proliferation of pine wood nematodes

To study the relationship between resistance to pine wilt disease and the migration or proliferation of pine wood nematodes (PWN) (Bursaphelenchus xylophilus), we conducted experiments using clonally-propagated Japanese black pines (Pinus thunbergii) with pre-evaluated individual resistance levels. Bark including the cortical resin canals—one of the main migration pathways of PWN—was removed by girdling, but neither the migration of PWN nor symptom development of pine wilt disease were inhibited by this treatment in non-resistant clones. Histological observations showed no significant differences in the lumen area or the number of cortical- and xylem- axial resin canals between resistant and susceptible clone groups from a half-sib family. A bioassay using methanol extracts from resistant and susceptible clones showed that extracts from both clones showed similar attractant effects to PWN, but neither had repellent effects. The resistant clones were multi-inoculated with PWN into three split points to mimic migration in the stem. The proportion of damaged plants was not significantly different from that in single-inoculated plants (control). In this experiment, the number of PWN detected from partially-damaged plants was much higher than that from non-damaged plants. An inoculation test using stem cuttings showed that the population of PWN increased in susceptible cuttings at 1–20 days after inoculation (dai), while it remained unchanged or gradually decreased in resistant cuttings. These findings suggested that the factors contributing to resistance were associated with inhibiting the proliferation of PWN, rather than inhibiting their migration.     

两种分子技术检测松木中松材线虫的效果评价

为更好地解决当前松材线虫Bursaphelenchus xylophilus检测工作中频现的假阴性问题,通过不同分子技术检测效果的比较评价,摸索出了一套可提高松材线虫检测准确性的技术体系。针对早期感病松木的低线虫量,建立了线虫的最大量分离方法和基于rDNA ITS序列的分子检测体系,该体系可高效检测出单条松材线虫,准确率达93.75%。以来自浙江省不同疫区的96份松木样品为检测材料,比较了SCAR标记和ITS序列2种分子检测技术的阳性率。结果显示,通过首次PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率分别为52.08%和55.21%,SCAR标记的检测阳性率为30.21%;通过第二次或巢式PCR,ITS_Ⅰ和ITS_Ⅱ序列的检测阳性率提高到了97.92%和100.00%,SCAR标记的检测阳性率提高到了59.38%,表明基于ITS序列的检测阳性率明显高于SCAR标记,且通过第二次或巢式PCR方法可进一步提高检测灵敏度,降低假阴性率。因此,通过线虫的最大量分离并基于rDNA ITS序列的分子检测可明显提高检测准确性,更适用于松材线虫的常规检测。     

Comparison of the reproductive potential of <Emphasis Type="Italic">Monochamus galloprovincialis</Emphasis> on two pine species under laboratory conditions

Monochamus galloprovincialis Olivier (Coleoptera: Cerambycidae) is one of the vectors of the pinewood nematode Bursaphelenchus xylophilus (Steiner and Buhrer) Nickle (Nematoda: Parasitaphelenchidae), the causal agent of pine wilt in susceptible pine trees. It is therefore important to study both the biological and ecological characteristics of the beetle. In this study reproductive potentials of female beetles reared on two pine species were investigated under laboratory conditions and compared using fertility tables constructed for beetles reared on each tree species. Beetle population parameters were compared between tree species. The intrinsic rate of increase and associated population parameters of beetles reared on Pinus sylvestris L. were consistently higher than those reared on P. nigra Arnold. This result suggests that P. sylvestris has a significantly greater positive effect on the reproductive potential of M. galloprovincialis populations than does P. nigra.     

三株芽胞杆菌菌株对松材线虫低温存活与繁殖的影响

为明确细菌在松材线虫生态适应性中的作用,本研究选择与致病相关的松材线虫伴生细菌GD1、马尾松内生细菌GD2以及具有杀松材线虫活性的湿地松内生细菌NJSZ-13为试验对象,测定经这3株芽胞杆菌菌株3个浓度低温驯化10、15和20 d后,在冷冻条件下松材线虫强毒虫株AMA3、中毒虫株AA3和弱毒虫株YW4的存活率和繁殖量。结果表明:低温驯化15 d和20 d后3株菌株对不同毒力线虫的活力影响较驯化10 d后的更显著。在低温驯化15 d、-20℃冷冻处理1 h后,5×10⁶CFU/mL浓度菌株GD1处理下,虫株AMA3、AA3和YW4的存活率分别为77.22%、83.68%和84.26%,与对照差异显著;5×10⁵CFU/mL浓度菌株GD1处理下,虫株AMA3、AA3和YW4的存活率分别为75.76%、80.67%和81.50%,与对照差异显著。5×10⁶CFU/mL和5×10⁵CFU/mL浓度菌株GD2处理下,与GD1处理组结果相似,菌株NJSZ-13处理组则与菌株GD1和GD2的结果相反。低温驯化15...     

Further studies on the role of monoterpenes in pine host selection and oviposition ofThaumetopoea pityocampa

In a young mixed pine plantation (Pinus nigra andP. pinea) in central Italy, the influence of the pine needle monoterpenes (R)-(+)-limonene, (S)-(−)-limonene and (1S)-(−)-β-pinene on the oviposition activity of the pine processionary moth,Thaumetopoea pityocampa (Denis et Schiffermüller), was assessed. Two application methods were compared: spraying the trees with the substances emulsified with water, and setting up polyethylene dispensers from which the monoterpenes evaporated. During the moth’s flight period weekly treatments (at 500 mg of monoterpene /l water) were carried out and the evaporation from the polyethylene dispensers (each with 5 g of a single monoterpene) was checked on each occasion by weighing the dispensers. The GLM analysis did not uncover any difference between the methods of application or between the pine species.T. pityocampa laid a significantly (P=0.01) lower number of egg masses on the trees treated with (1S)-(−)-β-pinene than on those treated with the other substances and on the control trees. A ∼45% reduction in egg mass number was obtained on the pines treated with (1S)-(−)-β-pinene, independent of the level of infestation, method of application or pine species. (R)-(+)-limonene and (S)-(−)-limonene were not effective. http://www.phytoparasitica.org posting July 29, 2008.     

Effect of log seasonality on reproductive potential ofMonochamus galloprovincialis reared in scots pine logs under laboratory conditions

Insect reproduction may be affected by a number of factors including seasonal changes in larval or adult nutrition. The effect of season on the reproductive potential ofMonochamus galloprovincialis (Olivier) females reared inPinus sylvestris L. (Scots pine) logs was investigated by constructing fertility tables for each log that differed only in the season they were cut. Population parameters were compared among three seasonal cohorts. The intrinsic rate of increase and most of the associated population parameters of beetles that emerged from logs cut during spring were higher than for beetles emerged from summer and autumn logs. Slight differences were found between summer and autumn cohorts. We suggest that seasonal differences in the nutritional quality of logs caused differences inM. galloprovincialis survival and reproductive potential. http://www.phytoparasitica.org posting March 16, 2008.     

小麦籽粒营养物质和次生物质含量与其对玉米象抗性的关系

为明确生产栽培小麦品种对玉米象Sitophilus zeamais(Motschulsky)的抗性机制,采用选择性和非选择性试验法系统评估了9个小麦品种对玉米象的抗性,并分析了小麦籽粒营养物质和次生物质含量与其抗性的关系。结果显示,不同小麦品种对玉米象的抗性存在显著差异,其中陕麦139和西农88的不选择性和抗生性较弱,小偃22、周麦16和周麦11较强;玉米象成虫对西农165选择性最强,虫口数为152.00头,而卵至子代成虫发育历期最长,为35.06 d,子代成虫数、重量损失率和敏感系数均较低,分别为19.67头、5.37%和8.30;对周麦18选择性最差,敏感性居中。玉米象成虫对不同小麦品种的选择性与各物质含量,以及总酚、蛋白质含量与各抗性指标均无显著相关性,但阿魏酸含量与子代成虫数、敏感系数及重量损失率显著负相关;可溶性糖含量与子代成虫数、敏感系数及重量损失率显著正相关,与卵至子代成虫发育历期显著负相关。表明小麦品种对玉米象的抗生性与其籽粒中阿魏酸和可溶性糖含量相关,阿魏酸含量越高、可溶性糖含量越低,小麦对玉米象的抗性越强。     

Incidence and distribution of Heterobasidion and Armillaria and their influence on canopy gap formation in unmanaged mountain pine forests in the Swiss Alps

Various disturbance factors on different spatial scales can lead to the creation of canopy gaps in forest ecosystems. In this study, we investigated the role of root rot fungi in the formation of canopy gaps in the Swiss National Park in the Central Alps. Dying or recently dead mountain pine (Pinus mugo subsp. uncinata trees (n = 172) and saplings (n = 192) from 42 canopy gaps were assessed for Armillaria and Annosum root rot. Heterobasidion annosum s.str. proved to be the dominant pathogen and was isolated from 49% of the trees and 64% of the saplings. Armillaria was found on 13% of the trees and 20% of the saplings. Three Armillaria species, A. borealis, A. cepistipes, and A. ostoyae, were identified. Armillaria ostoyae was the most frequent species, accounting for 72% of all Armillaria isolates. A total of 31 (74%) gaps were associated with H. annosum, and six (14%) with A. ostoyae. The remaining gaps were either associated with both pathogens (7%) or with other, unknown, factors (5%). Our findings suggest that the two pathogenic fungi, H. annosum s.str. and A. ostoyae, are the main reason for the large-scale mortality of mountain pines and the creation of canopy gaps in high elevation forests of the Swiss National Park.     

Effects of fertilization with ammonium sulphate and potassium sulphate on the development of Sphaeropsis sapinea in Corsican pine

Potted plants ofPinus nigra subsp.laricio were fertilized with ammonium sulphate (AS) and potassium sulphate (PS) each year during 3 consecutive years. The then 5-year-old plants were artificially inoculated withSphaeropsis sapinea. The fungus caused much bigger bark necroses in plants that had been fertilized with AS, than in plants fertilized with PS. It is concluded thatP. nigra growing in poor sandy soil becomes more susceptible toS. sapinea when fertilized with an excess of AS. This may explain partly the increased incidence ofS. sapinea inP. nigra stands in areas with a high rate of atmospheric NH₄-deposition in the Netherlands.     

Characterization of two races of the stem and bulb nematode (Ditylenchus dipsaci) in Israel

Two distinct races ofDitylenchus dipsaci in Israel were identified: one, which infects and damages onion and garlic, reproduces on pea, but does not infect phalaris grass; and a second, which infects and damages phalaris—and, probably, also ‘Saia’ oats—but fails to infect onion and garlic. A new ‘garlic’ race of the nematode does not appear to have been introduced into Israel together with the ‘Lavinia’ garlic cultivar, as previously speculated, but rather the introduced Lavinia clone is highly susceptible to the existing ‘onion and garlic’ race ofD. dipsaci.     

Mechanisms of resistance in cotton to whitefly (<Emphasis Type="Italic">Bemisia tabaci</Emphasis>): Tolerance

Screen house experiments were conducted under no choice conditions, to determine the tolerance, a mechanism of resistance, in cotton under a population pressure of 150 and 300 pairs of whiteflies Bemisia tabaci (Gennadius) (Homoptera: Aleyrodidae). Twelve cotton genotypes, viz., four resistant (NHH 44, Supriya, LD694 and PA 183), four susceptible (F 846, IS-376/4/1/20/72, LK 861 and RS 2013) and four with moderate reaction (T x Maroon 2-78, MRC 6304Bt, RS 2098 and CNH 911) to B. tabaci were assessed in terms of decrease in plant height, number of nodes and chlorophyll content (a, b and total) from control. The study revealed that all three characters are important to identify tolerance. A strong tolerance mechanism is operating in NHH 44. The genotypes LD 694 and PA 183 were rated as moderately tolerant, however, IS 376/4/1/20/72, Tx Maroon 2-78, MRC 6304Bt, F 846 and CNH 911 exhibited no tolerance mechanism and were categorized as the most susceptible genotypes.     

High resistance to rice yellow mottle virus in two cultivated rice cultivars is correlated with failure of cell to cell movement

Rice yellow mottle virus (RYMV) accumulation in protoplasts and whole plants was investigated in two highly resistant cultivars, Tog5681 (Oryza glaberrima) and Gigante (Oryza sativa). Three susceptible cultivars, i.e. one O. glaberrima Tog5673 and two O. sativa (IR64, Ac. 2428), and a partially resistant cultivar (Azucena) were used as control. After inoculation, accumulation of coat protein (CP) and viral RNA were monitored on protoplasts, inoculated leaves, sheaths of inoculated leaves and newly infected leaves by serological and Northern blot analysis. Viral RNA accumulated to a similar extent in protoplasts from all cultivars studied. In contrast, three distinct in planta behaviors were noted. In susceptible plants (IR64, Tog5673 and Ac. 2428), there was high CP and RNA accumulation at 5 d.p.i. in whole plants, suggesting that cell to cell and vascular movements occurred before 5 d.p.i. in inoculated leaves. The second behavior concerned Azucena, which showed a delay (around 7 d.p.i.) of viral accumulation in inoculated leaves. The third behavior involved the highly resistant cultivars Tog5681 and Gigante. CP and viral RNA were not detected in these cultivars. The comparison of viral accumulation in protoplasts and plants suggested that resistance of the highly resistant cultivars Tog5681 (O. glaberrima) and Gigante (O. sativa) was not due to the inhibition of virus replication but rather to the failure of cell to cell movement.     

Relationships between Weather Variables, Airborne Spore Concentrations and Severity of Leaf Blight of Garlic Caused by Stemphylium vesicarium in Spain

Ascospores and conidia released into the air were recorded around plots on which garlic debris infected by Stemphylium vesicarium were fixed onto the soil surface. Symptoms in garlic trap plots located in the vicinity of infected debris, started in March and developed during April–May to reach disease incidence close to 100%, final disease severity values being lower in 1993 and 1995 than in 1994 and 1996. Whereas daily concentrations of ascospores were rather erratic, with 30% of captures between 0 and 6 h, conidia showed a daily periodicity with highest concentrations between 12 and 18 h, with a pronounced peak between 14 and 16 h, and lowest values at night. Ascospore release occurred mainly in February and March. It coincided with rainfall periods, 14 h with vapour pressure deficit 5 mb and solar radiation <145 W m^–2 on the current day of the capture. In contrast, greatest captures of conidia started in late April and were prevalent in May, and were associated with rainfall in days previous to the capture in which rather high temperature occurred and solar radiation was 109–345 W m^–2. Among the weather variables considered, rainfall appeared directly related to the aerial concentration of ascospores and conidia. The role of relative humidity seemed essential when rainfall did not occur. There was a relationship between conidia concentration in the air and number of hours with temperature in the range 12–21 °C. Ascospore production was not essential for infections to take place, since primary infection from conidia may occur and disease can develop from them readily.     

Mode of transmission and morphological structures of two <Emphasis Type="Italic">Eremothecium</Emphasis> species between <Emphasis Type="Italic">Riptortus pedestris</Emphasis> and soybean

Eremothecium ashbyi and E. coryli were present in the maxillary and mandibular stylet pouches of the contaminated heads of the true bug Riptortus pedestris collected during a field survey, 2007. Ascospores of E. ashbyi and E. coryli were taken up in the stylet pouch when R. pedestris fed on soybean seeds infected with the respective fungus. In the case of E. ashbyi, the ascospores had subsequently germinated and were recognized as large masses of mycelia in the stylet pouch. In contrast, masses of E. coryli including buds of irregular size and shape were recognized about 3 days after. These results proved that these yeasts are taken up and become lodged in the stylet pouch when the insect feeds on infected host plants. In stylet sheaths stained with erythrosine, E. ashbyi was found as fragments of mycelia and E. coryli was found as small buds. These observations yield evidence to confirm that E. ashbyi and E. coryli are transmitted as fragments of mycelia and as small buds, respectively, after the insect has fed on infected host plants.     

Local and systemic resistance to fungal pathogens triggered by an AVR9-mediated hypersensitive response in tomato and oilseed rape carrying the Cf-9 resistance gene

Tomato and transgenic oilseed rape plants expressing the Cf-9 resistance gene develop a hypersensitive response (HR) after injection of the corresponding Avr9 gene product. It was investigated whether induction of a HR conferred resistance to different fungal pathogens in tomato and oilseed rape. Induction of an AVR9 mediated HR at the pathogen infection site delayed the development of the biotrophs Oidium lycopersicum in tomato and Erysiphe polygoni in oilseed rape, but enhanced the development of the necrotrophs Botrytis cinerea and Alternaria solani in tomato and Sclerotinia sclerotiorum in oilseed rape. Interestingly, delayed fungal disease development was observed in plant tissues surrounding the HR lesion regardless of whether a necrotrophic or biotrophic pathogen was used. In tomato, AVR9 injection induced systemic expression of PR1, PR2 and PR3 defence genes but did not induce systemic resistance to O. lycopersicum, B. cinerea or A. solani. In oilseed rape, AVR9 injection temporarily induced systemic resistance to Leptosphaeria maculans and E. polygoni, but did not induce detectable systemic expression of PR1, PR2 or Cxc750. These results give new insights into the potential uses of an induced HR to engineer disease resistance.     

几种生化物质与西瓜抗蚜性的相关性

为了初步探讨西瓜抗蚜性的营养与次生代谢物质防御效应,采用分光光度法测定并分析了几种生化物质与西瓜抗蚜性的相关性。结果表明,抗性品种黑皮、绿美人、黑美人和惠兰的游离氨基酸、脯氨酸和可溶性氮含量分别为2.22~2.27、0.23~0.26和1.88~3.03 mg/g,显著低于感性品种金美人、小玲、甜美人和花绿的3.24~3.83、0.67~0.87和5.26~5.82 mg/g;抗性品种的糖/氮比为5.38~7.94,显著高于感性品种;抗、感品种间可溶性糖和维生素C(Vc)含量无显著差异;瓜蚜为害4 d后的抗性品种叶组织中的总酚和单宁酸含量显著增加,感性品种显著降低;丙二醛含量增加幅度显著高于抗性品种。研究表明,西瓜品种抗蚜性与糖/氮比及总酚、单宁酸含量均呈显著正相关,与可溶性氮、游离氨基酸、脯氨酸及丙二醛含量均呈显著负相关,但与可溶性糖和Vc含量无显著相关性。     

基于EPG技术分析杭菊两主栽品种对三种菊蚜抗性及其相关抗性物质

为比较杭白菊主栽品种早、晚小洋菊对菊小长管蚜Macrosiphoniella sanborni、棉蚜Aphis gossypii、桃蚜Myzus persicae的抗性,通过刺吸电位技术(electrical penetration graph,EPG)研究菊蚜在菊顶叶上刺吸行为,并检测了菊顶叶主要化合物。结果表明:在晚小洋菊上,菊小长管蚜的E1、E2波平均持续时间4.31、3.47 min,分别短于在早小洋菊上的4.63、3.75 min;棉蚜、桃蚜的E1、E2波平均持续时间分别为4.32、4.72 min和4.92、4.64 min;3种蚜虫的平均刺探次数均大于在早小洋菊上的平均刺探次数。聚类分析结果显示,2个主栽品种之间抗性存在差异。早小洋菊顶叶平均可溶性糖和可溶性蛋白含量分别为2.71 mg/g和25.36 mg/g,均高于晚小洋菊;早小洋菊顶叶总酚含量为0.24 mg/g,显著低于晚小洋菊;早、晚小洋菊总黄酮含量分别为3.46 mg/g和3.37 mg/g。早小洋菊顶叶可溶性糖、可溶性蛋白含量基本上与每种菊蚜的E1、E2波持续时间显著正相关,总酚含量、总黄酮含量与每种菊蚜的E1或E2波持续时间显著负相关。推测早小洋菊对于菊长管蚜的抗性稍弱于晚小洋菊,2种杭白菊对棉蚜或桃蚜的抗性相当,且菊叶中的这4种物质含量与抗蚜性相关。     

A comparative study of resistance to powdery mildew in wild emmer wheat in the seedling and adult plant stage

An isometric virus was isolated from cucumber plants growing in a plastic house in Crete and showing stunting and bright yellow mosaic of the leaves. Based on host range, properties in crude sap, behaviour during purification, electron microscopy and serology, the virus was identified as an isolate of artichoke yellow ringspot nepovirus. Ecological data corroborate transmission of the virus via the soil.Samenvatting Uit komkommerplanten in plastic-foliekassen op Kreta werd een bolvormig virus geïsoleerd; de aangetaste komkommerplanten vertoonden dwerggroei en helder geel mozaïek op de bladeren. Gebaseerd op de resultaten verkregen uit onderzoek met het virus naar de waardplantenreeks, de eigenschappen in perssap, zuivering, elektronen-microscopie en serologie kon het virus worden geïdentificeerd als een strain van het artichoke yellow ringspot nepovirus. Waarnemingen op het gebied van de ecologie wijzen op overdracht van het virus via de grond.