Electrical penetration graphs from Nilaparvata lugens on resistant and susceptible rice varieties

The feeding behaviour of Nilaparvata lugens (Stal.) was electronically monitored on three rice varieties showing different levels of resistance in the Philippines, over an 8 h period, using a DC amplifier and a video-assisted observation method to simultaneously record honeydew production.Six electrical penetration graph (EPG) patterns are described by their frequency and amplitude characteristics. Patterns 4 and 5 were associated with the production of two different types of honeydew; basic honeydew containing amino acids was produced during Pattern 4 and acidic honeydew with no amino acids was produced during Pattern 5. These patterns are thought to indicate phloem and xylem ingestion by N. lugens respectively.Insects produced shorter periods of Pattern 4 on moderately resistant IR46 and resistant IR62 rice varieties than on susceptible IR22, although the time taken to produce the first period of Pattern 4 from the start of EPG recording did not differ between varieties. This suggests that N. lugens has the ability to initiate phloem ingestion on all three varieties, but that resistance mechanisms in IR46 and IR62 prevent sustained phloem ingestion, at least within the first 8 h of contact.

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Résumé Le comportement alimentaire de N. lugens Stahl a été enregistré életroniquement pendant 8 heures aux Philippines sur 3 variétés de riz présentant différents degrés de résistance, en utilisant un amplificateur DC associé à un système vidéo pour enregistrer simultanément la production de miellat.Six types d'enregistrements de pénétration (EPG) sont décrits avec leurs caractéristiques de fréquence et d'amplitude. Les types 4 et 5 sont associés à la production de 2 types différents de miellat; le miellat de base contenant des acides aminés est produit pendant le type 4, et le miellat acide sans acides aminés est produit pendant le type 5. On considére que ces types indiquent respectivement l'ingestion de phloème et xylème par N. lugens.Les périodes de type 4 ont été plus courtes sur les variétés modérément résistante IR46 et résistante IR62 que sur la variété sensible IR22, bien que le temps passé à produire la première période de type 4 depuis le début des enregistrements des EPG ne diffère pas suivant les variétés. Ceci laisse penser que N. lugens a la possibilité de commencer la consommation de phloème sur les trois variétés, mais que les mécanismes de résistance chez IR46 et IR62 empêchent une ingestion soutenue de phloème, au moins pendant les 8 premières heures de contact.

     

Variation in feeding behaviour of Nilaparvata lugens on resistant and susceptible rice varieties

The feeding behaviour of Nilaparavata lugens was monitored on three rice varieties showing different levels of resistance in the Philippines, using a video-assisted observation method. N. lugens made more frequent, shorter probes on the moderately resistant IR46 and resistant IR62 rice varieties than on the susceptible IR22. Honeydew production was significantly lower on the resistant varieties though insect weight gains in 24 h were similar on IR46 and IR22, both being significantly greater than on the highly resistant variety.Population development, growth index and damage ratings were low on IR62 indicating antibiosis and/or non preference. When IR46 plants were infested as seedlings population increase, growth index and damage ratings were similar to those on the susceptible IR22. When infested at a later stage of plant growth the damage rating showed a moderate level of resistance though some population development was maintained, indicating antibiosis and tolerance. N. lugens started probing less frequently after surface exploration on both resistant varieties than on IR22 suggesting the presence of a resistance factor associated with the surface waxes of these varieties.

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Résumé Le comportement alimentaire de Nilaparvata lugens sur variétés de riz, sensible (IR22), partiellement résistante (IR46) et fortement résistante (IR62), a été contrôlé avec une méthode associant la vidéo à l'observation. N. lugens faisait des piqûres plus fréquentes et plus brèves sur IR46 et IR62, que sur la variété sensible. La production de miellat était significativement plus faible sur les variétés résistantes, bien que les gains de poids des insectes aient été les mêmes en 24 h sur IR46 et IR22, les deux étant significativement supérieurs à celui sur IR62.La croissance de la population, l'indice de croissance et le taux de dégâts étaient tous plus faibles sur IR62, ce qui révèle une antibiose et/ou une absence de préférence. Quand la contamination des IR46 a au lieu au stade semis, la croissance de population, l'indice de croissance et le taux de dégâts étaient semblables à ceux de la variété sensible IR22. Quand la contamination avait lieu à un stade ultérieur, le laux de dégâts révélait un niveau modéré de résistance bien qu'une certaine croissance de population se soit maintenue, ce qui révèle antibiose et tolérance.Après exploration de la surface des feuilles des deux variétés résistantes, N. lugens sondait moins fréquemment que sur IR22, ce qui laisse présumer un facteur de résistance associé aux cires superficielles de ces variétés.

     

水稻品种中抗褐飞虱抗原次生物质的分析

借助高效液相色谱(HPLC)技术,研究了130份对褐飞虱生物型Ⅱ具有不同抗性水平的水稻样品(26个品种)中13个次生物质含量(峰面积)的差异;运用主成分分析和多元回归分析,建立了水稻品种抗性级别的预测模型：Y=3．4593-0．02491X1+0．08475X2-0．04227X8+0．1174X12结果表明,水稻的抗性水平与峰面积值之间极显著相关(r^2=0．84,P<0．01),峰1、峰2、峰8、峰12对应的次生化合物是影响水稻对褐飞虱生物型Ⅱ抗性水平的主要抗原次生化合物;水稻品种中起抗虫作用的抗原次生物质不止一种,而是几种的组合,而且它们对水稻抗虫性的贡献权重是不完全相同的．     

Responses by the brown planthopper,Nilaparvata lugens,to conspecific density on resistant and susceptible rice varieties

This study examines the nature of intraspecific interactions among Nilaparvata lugens (Stål) (Hemiptera: Delphacidae) planthoppers feeding on resistant and susceptible rice varieties. Planthopper nymphs produced less honeydew and gained less weight when feeding on rice variety IR62 (resistant) compared to susceptible rice varieties. A series of bioassays was conducted that varied N. lugens nymph densities on IR62 and IR22 (susceptible). Increasing nymph density facilitated feeding by conspecifics; however, intraspecific competition increased mortality of nymphs on IR62 (but rarely on IR22). Furthermore, nymph weights declined with increasing conspecific density on IR22, and the effects were weak on IR62. More female nymphs than males survived on IR62 but this was not affected by density. Nitrogenous fertilizer increased competition among N. lugens on young plants of IR22, but not on IR62. Results indicate that nymphs have a low efficiency in accessing resources when feeding on IR62, even where the plants have received fertilizer. Female‐biased survival and biomass compensation for mortality may promote population recovery after development on the resistant plant and accelerate adaptation to the resistant variety.     

抗性水稻品种对褐飞虱和白背飞虱种群参数的影响

在半自然状态下 ,用生命表方法初步比较了褐飞虱Nilaparvatalugens和白背飞虱Sogatellafurcif era在 4个抗褐飞虱水稻品种ASD7,IR3 6,JX89和Mudgo上的种群参数。初步的结果表明 ,白背飞虱在 4个抗褐飞虱水稻品种上具有较高的繁殖率 ,较短的发育时间和较大的种群增长。对褐飞虱各种群参数影响最大的是JX89,对白背飞虱影响最大的是Mudgo。与感性品种TN1比较 ,这些抗褐飞虱的水稻品种对 2种稻飞虱均具有不同程度的抗性。这些结果说明 ,抗褐飞虱水稻品种对白背飞虱同样具有一定的抗性 ,但白背飞虱对这种抗性具有较强的适应性 ,如果在一个地区长期推广抗褐飞虱的水稻品种 ,势必会导致白背飞虱的大发生     

Quantitative genetic analysis of biotypes of the brown planthopper Nilaparvata lugens: heritability of virulence to resistant rice varieties

The virulence of Nilaparvata lugens Stål (Homoptera: Delphacidae) to a resistant variety of rice, Oryza sativa L., is suggested to be under polygenic control. To determine whether the virulence of N. lugens is a quantitative character that has continuous distribution or a threshold character that has a few discontinuous phenotypic forms and is determined by some underlying continuous variable, I examined the frequency distribution of honeydew excretion that has been used as a measure of ability of attacking the rice plants, and the relationship between honeydew excretion and the oviposition rate of N. lugens females using a japonica rice line Saikai 190 which has a resistance gene Bph 1. The frequency distribution in honeydew excretion significantly differed from the normal distribution, but two distributions below and above 10 mg honeydew excretion for two days did not significantly deviate from normality, suggesting a bimodal distribution. There were significant differences in the proportion of females ovipositing and the number of eggs deposited between the female groups that excreted 0–10 mg honeydew and more than 20 mg honeydew. Within these female groups, however, the reproductive performances were not different. Thus, virulence of N. lugens can be analyzed as a threshold character that has two distinct phenotypes, virulent and avirulent. I estimated the heritabilities of the virulence of N. lugens by parent-offspring regression using the percentage of virulent females in a full-sib family. The estimates of heritability were 0.41 on Saikai 190 and 0.55 on ASD7, carrying the bph 2 gene. These results show that the N. lugens population has substantial genetic variation in virulence. The regression coefficients on female and male parents were similar, suggesting similar genetic contributions by both parents. When the parental families were examined on Saikai 190 and the progeny on ASD7, the regression coefficient approached zero. There may be a low genetic correlation between virulences to the two varieties.     

温度对褐飞虱抗/感吡虫啉品系种群生长的影响

与敏感性品系相比 ,经过连续 1 4代筛选的褐飞虱抗性品系对吡虫啉具有 48.5倍的抗性。通过构建不同温度下敏感品系和抗性品系的实验种群生命表 ,发现不适温度对 2个品系的生长发育和繁殖都有明显的影响 ,而且这种影响在 2个品系间存在差异。高温 ( 3 2℃ )和低温 ( 1 8℃ )导致 2个品系若虫存活率、羽化率、交配率、产卵量和孵化率明显下降。在 2 5℃ ,3 2℃和 1 8℃下 ,敏感品系的种群趋势指数分别为 1 64 3 ,2 6 7和 5 8 5 ,抗性品系的相应值分别为 81 4,9 5和 2 3 0。     

Genome-wide analysis of Nilaparvata lugens nymphal responses to high-density and low-quality rice hosts

The brown planthopper （BPH） Nilaparvata lugens is an economically impor- tant pest on rice plants. In this study, the higher population density and yellow-ripe stage of rice plants were used to construct adverse survival conditions （ASC） against BPH nymphs. Simultaneously, the low population density and tillering stage of rice plants were used to establish a suitable survival condition （SSC） as a control. Solexa/Illumina sequencing was used to identify genes of BPH nymphs responding to ASC. Significantly longer duration development of BPH nymphs and significantly lower brachypterous ratio of BPH adults were observed by ASC compared with SSC. A total of 2 544 differentially expressed genes （DEGs） were obtained and analyzed by BLASTx, Gene Ontology and KEGG Orthology. Gene ontology analysis revealed that the DEGs were mainly involved in categories of cell, cell part, cellular process, binding, catalytic, organelle and metabolic processes. 1138 DEGs having enzyme commission numbers were assigned to different metabolic pathways. The largest clusters were neurodegenerative diseases （137, 12.0%）, followed by carbohy- drate metabolism （113, 9.9%）, amino acid metabolism （94, 8.3%）, nucleotide metabolism （76, 6.7%）, energy metabolism （64, 5.6%）, translation （60, 5.3%）, lipid metabolism （58, 5.1%）, and folding, sorting and degradation （52, 4.6%）. Expressing profile of 11 DEGs during eight nymphal developmental stages of BPH were analyzed by quantitative real- time polymerase chain reaction. The 11 genes exhibited differential expression between ASC and SSC during at least one developmental stage. The DEGs identified in this study provide molecular proof of how BPH reconfigures its gene expression profile to adapt to overcrowding and low-quality hosts.     

十六个水稻品种(系)对褐飞虱的抗虫性评价

本文通过苗期抗性鉴定、田间抗虫性鉴定、稻田节肢动物功能团优势度比较及产量测定的方法来评价16个水稻品种(系)对褐飞虱Nilaparvata lugens(Stl)的抗性。结果表明,有12个品种(系)对褐飞虱表现为抗级,2个品系表现为中抗。这14个抗虫-中抗的品种(系)在成株期也表现一定的抗虫性,且与对照TN1之间差异显著。本试验条件下,广占63-3S/华恢15、广两优106、Y58S/华15、广两优476、广占63-4S/华恢15这5个品种(系)不仅抗虫性好,而且产量也较高,分别比TN1和主栽品种扬两优6号高63.49%～66.57%和3.47%～11.57%,具有广泛的开发应用前景。     

褐飞虱侵害后不同水稻品种根及地上部氮、磷、钾含量的变化

利用浓硫酸-过氧化氢消煮法,研究了不同耐、感虫品种水稻分蘖期在褐飞虱侵害胁迫后根及地上部间营养成分含量的变化情况。结果表明：褐飞虱侵害协优963后3天,根及地上部N、P、K含量、6天根及地上部N含量、9天地上部N含量在60、120头/株侵害后变化不明显;6天根及地上部P含量、6天根K含量、9天地上部P含量、9天根及地上部K含量在120头/株侵害后显著下降,60头/株侵害后变化不明显;6天地上部K含量、9天根N含量在60、120头/株侵害后均显著下降。对于协优63,3天地上部N含量、3天根及地上部P含量、6天及9天地上部N、P含量在60、120头/株侵害后变化不明显;6天根N、P含量、根及地上部K含量、9天根N含量在120头/株侵害后显著下降,60头/株侵害后下降不明显;3天、9天根及地上部K含量、9天根部P含量在60、120头/株侵害后均显著下降。表明不同水稻品种体内不同营养物质含量在褐飞虱侵害后变化不同,协优63较协优963敏感;K含量变化最明显,其次为P,最后为N;而且根比地上部对褐飞虱的反应敏感。     

Photosynthesis and translocation of assimilates in rice plants following phloem feeding by the planthopper Nilaparvata lugens (Homoptera: Delphacidae)

Experiments were conducted to measure the effect of feeding by the planthopper Nilaparvata lugens (St?l) on photosynthesis and the translocation of assimilates in rice plants, Oryza sativa L. We used mature japonica rice plants and applied the 13CO2 feeding method to evaluate those physiological effects. The photosynthetic rate was suppressed by N. lugens infestation, especially at the lower leaf position, with rates 30% lower than that of control plants at the booting stage. Leaf nitrogen concentration in infested plants was also lower than that in control plants. After flowering, the chlorophyll content and total plant dry weight were reduced by N. lugens. Stem and sheath dry weights were significantly reduced by N. lugens infestation, whereas panicle dry weight was not affected. Little effect was found on disruption in translocation of assimilates, even when 13CO2 was supplied to the infested leaves. Results suggested that removal of assimilates and reduction in photosynthesis by N. lugens have the greatest effect on growth and yield of rice plants as compared with the disruption in the translocation of assimilates. Plant death can occur by N. lugens infestation if the amount of energy supplied is less than that required for tissue maintenance.     

Pesticide-induced susceptibility of rice to brown planthopper Nilaparvata lugens

The effects of rice plants treated with various pesticides (jingganmycin, bisultap and methamidophos) on feeding, survival rates and population growth of Nilaparvata lugens Stål (Homoptera: Delphacidae), susceptibility of the treated rice plants and amounts of free amino acids and sucrose were studied. Experiments indicated that the effects of the tested pesticides were dependent on nymphal age, pesticide and their dose and time after application. Jingganmycin at 75 g a.i. ha^–1 significantly increased the N. lugens population. Both jingganmycin and bisultap increased the survival rate of N. lugens nymphs. The feeding rate of the insects was also affected by the pesticide application, but the effect varied between nymphal age and time after application and lasted no longer than 15 d. Results clearly indicated that pesticide application increased the susceptibility of rice plants to N. lugens. Although the free amino acids in rice plants did not change with the pesticide treatments, the concentration of sucrose significantly decreased 5 d after application and the C/N ratio significantly decreased in jingganmycin treated plants 5 d and 10 d after application.     

褐飞虱侵害对不同抗性水平水稻根部吸收氮、磷、钾的影响

为了解褐飞虱Nilaparvata lugens侵害后水稻抗性水平与根系吸收氮（N）、磷（P）、钾（K）的关系,对不同抗性水平的水稻受褐飞虱侵害后的平均受害水平、植株功能损失指数及水稻吸收水培液营养元素N,P,K情况进行了研究。结果表明：不同水稻品种（TN1、协优63、协优963和超级培矮64S/E32）接种褐飞虱后,TN1受害最严重,协优63和超级培矮64S/E32 次之;受害最轻的是协优963,30头/株侵害后植株功能损失指数仅为0.661。40,60和80头/株侵害水稻后根系对K吸收下降程度最显著,其次为P,最后为N;且随褐飞虱侵害时间的延长（6 d, 9 d）影响愈显著。60头/株侵害TN1、协优63、协优963和超级培矮64S/E32后9 d,根系对K吸收下降率分别为164.11%,74.61%,55.16%和46.60%。由此可见,随水稻品种抗性水平的下降,接种褐飞虱后,水稻根部对水培液营养元素吸收下降程度愈显著。本研究结果可对深入阐明不同抗性水平水稻抗（耐）虫机制与根系吸收能力关系提供参考。     

褐飞虱对抗性水稻品种Mudgo个体致害性指标

在28±1℃下,观测了褐飞虱Nilaparvata lugens羽化24?h内短翅型雌成虫在敏感水稻品种TN1及抗性品种Mudgo上的寿命及体重增量。结果表明,取食抗性品种时,试虫的平均增重与寿命之间表现出明显的正相关。寿命在7天内的个体,存活期间平均体重增量为负值,而存活7天以上的个体则平均增重明显。进一步研究了初羽化的褐飞虱长翅型雌成虫在TN1及Mudgo上取食第1～4天的体重增量、寿命及产卵量,结果发现试虫在Mudgo上的这3个生物学参数之间存在明显正相关。其中,寿命及体重增量较好地反映出害虫个体对抗性品种的致害能力,可作为个体致害性指标。根据研究结果,提出将羽化后在Mudgo上存活7天以上或最初4天内体重增量大于0.1mg的雌成虫定为能够致害该抗性品种的个体。     

Survival rate and stylet penetration behavior of current Korean populations of the brown planthopper,Nilaparvata lugens,on resistant rice varieties

To understand the development of host plant resistance-breaking ability of the current BPH populations in Korea, we conducted nymphal survivorship tests and electrical penetration graph (EPG) studies on susceptible and resistant rice varieties with four different BPH populations, which were collected in the early 1980s (S-BPH) and in 2005, 2006, and 2007. The S-BPH had low survival rates on resistant rice varieties carrying either Bph1 or bph2. However, the current BPH populations have high resistance-breaking ability on the varieties with their elevated survival rates, whereas their survival rates were still low on the other resistant varieties, Gayabyeo (Bph1  the other unknown gene) and Rathu Heenati (Bph3). The EPG analysis also revealed that the ratio of BPH that could reach the phloem sap ingestion waveform (N4-b) within 15 h on the resistant rice varieties containing Bph1 or bph2 was higher in the current BPH populations (16.7–50%) than in the S-BPH population (0-4.2%). However, the pre-reaching time from the penetration start to the first N4-b waveform in the current BPH populations was significantly longer on resistant varieties (Bph1 or bph2) than on susceptible varieties. Furthermore, the total duration of N4-b waveform was significantly shorter on the resistant varieties.From these results, we suggest that, although the current BPHs collected in Korea have a high resistance-breaking ability through the increase of survival rate on resistant rice varieties carrying either Bph1 or bph2, they still have some difficulties feeding on the phloem sap of the resistant rice varieties.     

Characterization of phosphate esters in the shoots and roots of alfalfa plants susceptible and resistant to bacterial wilt

Acid-soluble phosphate esters were determined in extracts of plant material after a 24 h exposure of the roots of intact alfalfa plants to nutrient media labelled with³²P_i. Similar phosphate ester patterns were found in 2-, 3-, 8-, and 9-week-old plants with the exception of Gra-P which could be detected only in shoot extracts. However, phosphate ester levels differed in the shoots and roots. Whereas Fru-P₂, Glc-6-P, Fru-6-P, and adenine nucleotides were more abundant in the shoots, Grn-P and P-choline levels were higher in the roots. Certain differences in the levels of P-esters were also recorded between plants susceptible and resistant to bacterial wilt.     

水稻品种对褐飞虱的田间抗性及其生态学机制

评价了7个水稻品种(浙粳22、宜香845、中组14、秀水123、二优倍九、感虫对照品种TN1和抗虫对照品种IR36)在田间成株期对褐飞虱Nilaparvata lugens(Stl)的抗性表现,并初步分析了它们的抗性机制。结果表明,在田间网室人工诱发条件下,浙粳22、宜香845、秀水123和中组14等4个品种的抗性水平与抗虫品种IR36相同,均小于3级,表现为抗虫,而二优培九则与感虫品种TN1相同,表现为感虫。在自然发生条件下的水稻分蘖期至乳熟期,宜香845和中组14上褐飞虱数量仅为TN1上的7.84%～24.44%,秀水123和浙粳22则为TN1上的30.19%～60.27%。在4个抗性品种上褐飞虱的产卵量和卵孵化率明显减少,而若虫存活率仅为TN1上的20%～30%。与IR36相同,4个抗性品种上褐飞虱的若虫存活率和种群增长倍数均显著低于TN1上,说明他们对褐飞虱的抗性主要表现为抗生性。     

水稻活性成分的分离纯化及其对褐飞虱若虫的活性测定

水稻植株内的抗性次生物质是影响水稻生化抗性的主要因子之一。运用硅藻土系统溶剂分离法和硅胶柱层析分离方法分离纯化手段,从水稻(Oryza sativa L.)抗虫品种IR36甲醇粗提物中得到一种针状晶体,初步认为该晶体为一种糖甙黄酮类物质,通过该物质对褐飞虱Nilaparvata lugens(Stal)生物型Ⅱ若虫的活性测定,表明该种晶体对褐飞虱若虫的存活率、体重增加和羽化均具有显著的不良影响,不利于褐飞虱若虫的生长发育。     

Physiological responses of resistant and susceptible buffalograsses to Blissus occiduus (Hemiptera: Blissidae) feeding

The impact of Blissus occiduus Barber feeding on resistant ('Prestige') and susceptible ('378') buffalograsses, Buchlo? dactyloides (Nuttall) Engelmann, was evaluated through measurement of carbon exchange rate, light and carbon assimilation (A-C(i)) curves, chlorophyll a fluorescence, and nonstructural carbohydrates. No significant differences in carbon exchange rates were observed between infested and control plants for 378 at 5 and 10 d after infestation; however, at 20 d after chinch bug introduction, significant differences in carbon exchange rates between infested and control 378 plants were detected. Carbon exchange rates were similar between infested and control Prestige plants at 5,10, and 20 d after infestation, suggesting that resistant plants can allocate energy for recovery from chinch bug injury. Significant differences in the photochemical efficiency of photosystem II (PSII) and the apparent photosynthetic electron transport ratio were observed between infested and control 378 plants, whereas, no significant differences in the photochemical efficiency of PSII and the electron transport ratio were detected between control and infested Prestige plants. Blissus occiduus-infested 378 and Prestige plants consistently had similar or higher levels of nonstructural carbohydrates compared with their respective control plants. These data suggest that both resistant and susceptible buffalograsses increase levels of nonstructural carbohydrates in response to B. occiduus feeding. This research also suggests compensatory photosynthesis takes place in Prestige but not in 378.