Automated Counting of Rice Planthoppers in Paddy Fields Based on Image Processing

A quantitative survey of rice planthoppers in paddy fields is important to assess the population density and make forecasting decisions. Manual rice planthopper survey methods in paddy fields are time-consuming, fatiguing and tedious. This paper describes a handheld device for easily capturing planthopper images on rice stems and an automatic method for counting rice planthoppers based on image processing. The handheld device consists of a digital camera with WiFi, a smartphone and an extrendable pole. The surveyor can use the smartphone to control the camera, which is fixed on the front of the pole by WiFi, and to photograph planthoppers on rice stems. For the counting of planthoppers on rice stems, we adopt three layers of detection that involve the following：（a） the first layer of detection is an AdaBoost classifier based on Haar features;（b） the second layer of detection is a support vector machine（SVM） classifier based on histogram of oriented gradient（HOG） features;（c） the third layer of detection is the threshold judgment of the three features. We use this method to detect and count whiteback planthoppers（Sogatella furcifera） on rice plant images and achieve an 85.2% detection rate and a 9.6% false detection rate. The method is easy, rapid and accurate for the assessment of the population density of rice planthoppers in paddy fields.

The insecticide resistance in two planthoppers from three areas to three insecticides

Migrating insects brown planthopper(BPH),Nilaparvata lugens St(?)l andwhite-backed planthopper(WBPH),Sogatella furcifera Horvath are both mostharmful insects on rice in China.Chemical control is thought to be the best wayto manage them,but it may cause insecticide resistance.Methamidophos,buprofezin,and imidacloprid were the three insecticides often used.In 2000and 2001,resistance of the field populations collected from three areas,i.e.Dongtai,Jiangsu Province,Anqing,Anhui Province,and Guilin,GuangxiAutonomous Region,to the three insecticides was monitored.The toxicities ofthe three insecticides were determined with the female adult(3-5 d after e-mergence)and by the topical application method with the hand microapplicator.

Survey on Spawning Characteristics of Rice Planthoppers in Different Rice Varieties

The fecundity and spawning locations of rice planthoppers in several rice varieties were surveyed in the test. The results showed that spawning behavior of rice planthoppers obeyed obvious niche distribution; rice planthopper preferred to lay eggs in the leaf sheath with spawning mark distance more than 20 cm, while they liked to lay eggs in the leaves with spawning mark distance of 10 - 20 cm. The average fecundity in leaf sheaths was obviously higher than that in leaves, indicating that leaf sheaths were more suitable for spawning of rice planthoppers than leaves.

Relationship of egg parasitoids of rice planthoppers between rice and non-rice habitats

Egg parasitoids,Anagrus spp.and Oligositaspp.have been found to be important biologi-cal agents in suppressing rice planthoppers inAsian countries.However,most of these stud-ies have been confined to conditions within ricefields.In this study,field surveys were madeto study the floral and faunal diversity in vari-ous habitats surrounding rice fields.The riceand grass(Echinochloa spp.)plants were in-

Long range migration of rice planthoppers and associated meteorological conditions in Yulin,Ganzhou,and Huangyan

In recent years,it has been demonstrated that thelow-level jet streams(LLJs)developed in thesouth of the Meiyu(Baiu)front play a critical role inthe overseas migrations of the planthoppers intoJapan and Korea. However, the involement of theLLJs during the process of sequential displacements

The Influence of Transgenic cry1Ab/cry1Ac,cry1C and cry2A Rice on Non-Target Planthoppers and Their Main Predators Under Field Conditions

Transgenic Bt rice has been shown to be an effective means of controlling Lepidoptera pests of rice. However, the potential roles of transgenic rice on planthoppers and their predators need to be investigated before its commercialization. Population density, species dominance and population dynamics are important parameters of arthropods populations in field. So the impacts of three transgenic Bt rice strains expressing crylAb/crylAc, crylC and cry2A on population density, species dominance and population dynamics of three species of planthoppers （Nilaparvata lugens, Sogatella furcifera and Laodelphax striatellus） and their three main predators （ Cyrtorhinus lividipennis, Pirata subpiraticus and Theridium octomaculatum） were evaluated at three sites in Hubei Province, China, in the current study. The results showed that among three species of planthoppers, both in transgenic and non-transgenic rice field, the predominant species ofplanthoppers within phytophagous guild was S. furcifera at any site either growing season （46-50%）. Significantly higher population density ofN. lugens was observed in T2A-1 field relative to Minghui 63 field at Wuxue in 2010. The species dominance of predator, P. subpiraticus, in TT51 field was significantly higher than that in T 1 C-19 and T2A-1 fields in 2009 at Xiaogan site. Sampling date significantly influenced six arthropods except for P. subpiraticus in 2010. The interaction between rice strain^sampling date had no significant adverse effects on the population dynamics of three species of planthoppers and their predators, except for several individual species in 2009. The interaction among rice strain^sampling date^sampling site also had no significant effect on six arthropods except for S. furcifera in 2009. The results indicated that transgenic Bt rice expressing crylAb/crylAc, cry2A and crylC had no significant adverse effects on the population dynamics of three planthoppers and their predators in most investigated data and sampling site.

Automated detection and identification of white-backed planthoppers in paddy fields using image processing

A survey of the population densities of rice planthoppers is important for forecasting decisions and efficient control. Tra- ditional manual surveying of rice planthoppers is time-consuming, fatiguing, and subjective. A new three-layer detection method was proposed to detect and identify white-backed planthoppers （WBPHs, Sogatella furcifera （Horvath）） and their developmental stages using image processing. In the first two detection layers, we used an AdaBoost classifier that was trained on a histogram of oriented gradient （HOG） features and a support vector machine （SVM） classifier that was trained on Gabor and Local Binary Pattern （LBP） features to detect WBPHs and remove impurities. We achieved a detection rate of 85.6% and a false detection rate of 10.2%. In the third detection layer, a SVM classifier that was trained on the HOG features was used to identify the different developmental stages of the WBPHs, and we achieved an identification rate of 73.1%, a false identification rate of 23.3%, and a 5.6% false detection rate for the images without WBPHs. The proposed three-layer detection method is feasible and effective for the identification of different developmental stages of planthoppers on rice plants in paddy fields.

Gapless Genome Assembly of ZH8015 and Preliminary Multi-Omics Analysis to Investigate ZH8015's Responses Against Brown Planthopper Infestation

Accurate genomic information is essential for advancing genetic breeding research in specific rice varieties.This study presented a gapless genome assembly of the indica rice cultivar Zhonghui 8015(ZH8015)using Pac Bio HiFi,Hi-C,and ONT(Oxford Nanopore Technologies)ultra-long sequencing technologies,annotating 43037 gene structures.Subsequently,utilizing this genome along with transcriptomic and metabolomic techniques,we explored ZH8015's response to brown planthopper(BPH)infestation.Continuous transcriptomic sampling indicated significant changes in gene expression levels around 48 h after BPH feeding.Enrichment analysis revealed particularly significant alterations in genes related to reactive oxygen species scavenging and cell wall formation.Metabolomic results demonstrated marked increases in levels of several monosaccharides,which are components of the cell wall and dramatic changes in flavonoid contents.Omics association analysis identified differentially expressed genes associated with key metabolites,shedding light on ZH8015's response to BPH infestation.In summary,this study constructed a reliable genome sequence resource for ZH8015,and the preliminary multi-omics results will guide future insect-resistant breeding research.

Synergistic effects of the entomopathogenic fungus Isaria javanica and low doses of dinotefuran on the efficient control of the rice pest Sogatella furcifera

The rice planthopper,Sogatella furcifera,is a piercing-sucking insect pest of rice,Oryza sativa.It is responsible for significant crop yield losses,and has developed moderate to high resistance to several commonly used chemical insecticides.We investigated the effects of the insect fungal pathogen Isaria javanica,alone and in combination with the chemical insecticide dinotefuran,on S.furcifera under both laboratory and field conditions.Our results show that I.javanica displays high infection efficiency and mortality for different stages of S.furcifera,reducing adult survival,female oviposition and ovary development.Laboratory bioassays showed that the combined use of I.javanica with a low dose(4-16 mg L^(-1))of dinotefuran resulted in higher mortality in S.furcifera than the use of I.javanica or dinotefuran alone.The combined treatment also had more significant effects on several host enzymes,including superoxide dismutase,catalase,peroxidase,and prophenol oxidase activities.In field trials,I.javanica effectively suppressed populations of rice planthoppers to low levels(22-64%of the level in untreated plots).Additional field experiments showed synergistic effects,i.e.,enhanced efficiency,for the control of S.furcifera populations using the combination of a low dose of I.javanica(1×10^(4) conidia mL^(-1))and a low dose of dinotefuran(~4.8-19.2%of normal field use levels),with control effects of>90%and a population level under 50 insects per 100 hills at 3-14 days post-treatment.Our findings indicate that the entomogenous fungus I.javanica offers an attractive biological control addition as part of the integrated pest management(IPM)practices for the control of rice plant pests.

Various infection status and molecular evidence for horizontal transmission and recombination of Wolbachia and Cardinium among rice planthoppers and related species

Wolbachia and Cardinium are widely distributed and are considered important for their ability to disturb reproduction and affect other fitness-related traits of their hosts. By using multilocus sequence typing （MLST）, RFLP （restriction fragment length poly- morphism） and 16S ribosomal DNA gene sequencing methods, we extensively surveyed Wolbachia and Cardinium infection status of four predominant rice planthoppers and one kind of leafhopper in different rice fields. The results demonstrated that Sogatellafurcifera （Horv^ith） and Laodelphax striatellus （Fallrn） were infected with the same Wolbachia strain （wStri）, while Nilaparvata lugens （St^l） and its closely related species Nilaparvata muiri China were infected with two phylogeneticlly distant strains, wLug and wMui, respectively. Three new Wolbachia strains （provisionally named wMfasl, wMfas2 and wMfas3） were detected in the leafhopper Macrostelesfascifrons （Stal）. Only S. furcifera was co-infected with Cardinium, which indicated that the distribution of Cardinium in these rice planthop- pers was narrower than that of Wolbachia. Unambiguous intragenic recombination events among these Wolbachia strains and incongruent phylogenetic relationships show that the connections between different Wolbachia strains and hosts were more complex than we expected. These results suggest that horizontal transmission and host associated special- ization are two factors affecting Wolbachia and Cardinium infections among planthoppers and their related species.

Lipidomic analyses reveal enhanced lipolysis in planthoppers feeding on resistant host plants

The brown planthopper(BPH)(Nilaparvata lugens St?l)is a highly destructive pest that seriously damages rice(Oryza sativa L.)and causes severe yield losses.To better understand the physiological and metabolic mechanisms through which BPHs respond to resistant rice,we combined mass-spectrometry-based lipidomics with transcriptomic analysis and gene knockdown techniques to compare the lipidomes of BPHs feeding on either of the two resistant(NIL-Bph6 and NIL-Bph9)plants or a wild-type,BPH susceptible(9311)plant.Insects that were fed on resistant rice transformed triglyceride(TG)to phosphatidylcholine(PC)and digalactosyldiacylglycerol(DGDG),with these lipid classes showing significant alterations in fatty acid composition.Moreover,the insects that were fed on resistant rice were characterized by prominent expression changes in genes involved in lipid metabolism processes.Knockdown of the NlBmm gene,which encodes a lipase that regulates the mobilization of lipid reserves,significantly increased TG content and feeding performance of BPHs on resistant plants relative to dsGFP-injected BPHs.Our study provides the first detailed description of lipid changes in BPHs fed on resistant and susceptible rice genotypes.Results from BPHs fed on resistant rice plants reveal that these insects can accelerate TG mobilization to provide energy for cell proliferation,body maintenance,growth and oviposition.

The combination of NlMIP and Gαi/q coupled-receptor NlA10promotes abdominal vibration production in female Nilaparvata lugens(Stal)

For various sexually mature insects,including the brown planthopper(BPH,Nilaparvata lugens),the abdominal vibration(AV)signal is the initiation of the mating process,and itiscritical to the successofmating.Currently,there are few studies on the genetic and molecular mechanisms of AV regulation.Ourprevious AV-related transcriptomic study in female BPH identified myoinhibitory peptide(NlMIP)as a gene that potentially affects AV statusinfemales,buthow NlMIP affects AV status remains unknown.In this study,we confirmed that NlMIP regulates AV production and mating behavior in female BPH.When the RNAi knockdown efficiency of NlMIP was 59.00%,the probability of females producing AV and the mating rate in 1 h decreased by 38.89 and 61.11%,respectively.In addition,six mature peptides of NlMIP were synthesized and they were able to regulate AV production and mating behavior in females,with NlMIP2having the strongest effect.The A-family neuropeptide GPCR 10(NlA10)was found to be a potential receptor for NlMIP based on a phylogenetic tree analysis and the fact that NlMIP mature peptides effectively activated NlA10.After NlA10was knocked down,the probability of females producing AV and the mating rate in 1 h had reductions of 28.89 and 43.33%,respectively.When activated by NlMIP2,NlA10 coupled the Gαi/q signalling pathways,thereby inhibiting the downstream AC/cAMP/PKA,activating the PLC/Ca^(2+)/PKC signalling pathways and then activating MEK1/2 in a cascade to mediate the phosphorylation of ERK1/2,and finally regulating the AV of females.These results provide a basis for the prevention and control of the brown planthopper pest by disrupting female AV.

Studies on Insecticidal Activities and Active Ingredients of Stephania kwangsiensis Lo.

Insecticidal activities and active ingredients of Stephania kwangsiensis Lo. were studied for the first time. The results showed that all parts of S. kwangsiensis Lo. had contact activity against brown planthoppers, Nilaparvata lugens Stal, and the contact activity of methanol extract from root tubers was the highest, with a LD50 value being 1.5794 lag/female. l-roemerine was isolated from root tubers of S. kwangsiensis Lo. and identified, and it was the main active ingredient. l-roemerine had high contact toxicity to brown planthoppers, with a LD50 value being 0.0443 lag/female. Contact toxicity of l-roemerine to brown planthoppers was 7.48 times that of malathion, the convientional chemical insecticide used for controlling brown planthoppers.l-roemerine also had stomach poison activity against brown planthoppers.

基于主成分回归法的稻飞虱百穴虫量冠层光谱检测(英文)

[ Objective] Aiming at problems of early warning for occurrence of rice pests and dynamic monitoring of rice planthopper in field, a detection model for rice planthopper populations was established based on PCR with spectrum detection technology, r Method] Canopy reflectance data were collected using FieldSpeo 3 spectrometer in paddy field, and rice planthoppers populations in hundred hills were detected simultaneously. The sample size was 71, and there were 51 samples in the calibration set and 20 samples in the prediction set. Modeling band was 350 -1 139 nm, and the original spectra were pretreated by first order differential. [ Result] The correlation coefficient of measured values and predictive values was 0. 78, and the RMSEP was 161. [ Conlmion] Spectrum detection was able to be used in investigation and forecasting of rice planthoppere.

稻飞虱迁飞气象条件等级预报研究(英文)

By comparative analysis on the meteorological conditions and occurrences of rice planthoppers in Guilin of Guangxi during recent 5 years,the temperature,precipitation,wind direction,wind velocity,humidity were chosen as the factors that affected the migration of rice planthopper in Guilin.Thinking of meteorological conditions and injurious number in early period,regression analysis method was used to establish the work system for grade forecast of meteorological conditions that affected the migration of rice planthopper.The forecast factors and targets were divided as 5 grades in the work system.Using one-week's weather forecast conclusion that local observatory had published,whether the meteorological conditions and injurious number were favorable or not was analyzed synthetically.The meteorological conditions grades that affected the migration of rice planthopper for each day in future 1-7 days were predicted.This is a practical forecast work system and the forecast accuracy for each day is larger than 70%.The work system has positive function in the manufacture practice.

超级稻甬优6号褐飞虱种群繁殖速率的密度制约效应(英文)

[ Objective ] The paper was to study the density restriction effects of reproductive rate of rice brown planthopper （BPH） population in super indiea and japonica hybrid rice Yongyou 6. [Method] Different amounts of rice brown planthoppor were released to super indiea and japonica hybrid flee Yongyou 6 during fillering, booting and gain filling stage, respectively. The reproductive rate and variation dynamics of rice brown planthoppor in super rice was also investigated and analyzed. [Result] Tillering stage： the population quantity of rice brown planthoppor during booting and gain filling stage gradually increased with the increasing basal amount of its population during tillering stage; when the population density of rice brown planthopper during grain filling stage reached a certain limit （ 〉 250 head/cluster）, the population density during milking stage showed decrease trend with the increasing of such density; the development of rice brown planthoppor population in super rice Yongyou 6 showed the characteristics of fast in middle and slow in two ends. Booting stage： the development of rice brown planthopper pop- ulation from booting to gain filling stage increased with the increasing basal amount of density, and their correlation during milking stage was not significant; flee brown planthopper population still had high reproduction capacity in the middle and late stage of super rice, which had latent risk on rice yield. Gain filling stage ： when population density of rice brown planthopper 〈 70 head/cluster, the development of it population increased with the increase of basal amount of density, which had great threat to yield. [ Conclusion] The result provided theoretical basis for establishment and improvement of monitoring early warning system, cost-ef- fective control of occurrence and damage of rice brown planthopper, and improvement of the sustained control ability against rice brown planthopper.

Induced defense responses in rice plants against small brown planthopper infestation

The small brown planthopper(SBPH), Laodelphax striatellus Fallén(Homoptera: Delphacidae), is a serious pest of rice(Oryza sativa L.) in China. To understand the mechanisms of rice resistance to SBPH, defense response genes and related defense enzymes were examined in resistant and susceptible rice varieties in response to SBPH infestation. The salicylic acid(SA) synthesis-related genes phenylalanine ammonia-lyase(PAL), NPR1, EDS1 and PAD4 were induced rapidly and to a much higher level in the resistant variety Kasalath than in the susceptible cultivar Wuyujing 3 in response to SBPH infestation. The expression level of PAL in the Kasalath rice at 12 h post-infestation(hpi) increased 7.52-fold compared with the un-infested control, and the expression level in Kasalath was 49.63, 87.18, 57.36 and 75.06 times greater than that in Wuyujing 3 at 24, 36, 48 and 72 hpi, respectively. However, the transcriptional levels of the jasmonic acid(JA) synthesis-related genes LOX and AOS2 in resistant Kasalath were significantly lower than in susceptible Wuyujing 3 at 24, 36, 48 and 72 hpi. The activities of the defense enzymes PAL, peroxidase(POD), and polyphenol oxidase(PPO) increased remarkably in Kasalath in response to SBPH infestation, and were closely correlated with the PAL gene transcript level. Our results indicated that the SA signaling pathway was activated in the resistant Kasalath rice variety in response to SBPH infestation and that the gene PAL played a considerable role in the resistance to SBPH.

Development and Identification of Introgression Lines from Cross of Oryza sativa and Oryza minuta

Introgression line population is effectively used in mapping quantitative trait loci（QTLs）,identifying favorable genes,discovering hidden genetic variation,evaluating the action or interaction of QTLs in multiple conditions and providing the favorable experimental materials for plant breeding and genetic research.In this study,an advanced backcross and consecutive selfing strategy was used to develop introgression lines（ILs）,which derived from an accession of Oryza minuta（accession No.101133） with BBCC genome,as the donor,and an elite indica cultivar IR24（O.sativa）,as the recipient.Introgression segments from O.minuta were screened using 164 polymorphic simple sequence repeat（SSR） markers in the genome of each IL.Introgressed segments carried by 131 ILs covered the whole O.sativa genome.The average number of homozygous O.minuta segments per introgression line was about 9.99.The average length of introgressed segments was approximate 14.78 cM,and about 79.64% of these segments had sizes less than 20 cM.In the genome of each introgression line,the O.minuta chromosomal segments harbored chromosomal fragments of O.sativa ranging from 1.15% to 27.6%,with an overall average of 8.57%.At each locus,the ratio of substitution of O.minuta alleles had a range of 1.5% 25.2%,with an average of 8.3%.Based on the evaluation of the phenotype of these ILs,a wide range of alterations in morphological and yield-related traits were found.After inoculation,ILs 41,11 and 7 showed high resistance to bacterial blight,brown planthopper and whitebacked planthopper,respectively.These O.minuta-O.sativa ILs will serve as genetic materials for identifying and using favorable genes from O.minuta.

Pyramiding blast,bacterial blight and brown planthopper resistance genes in rice restorer lines

Rice blast, bacterial blight （BB） and brown planthopper （BPH） are the three main pests of rice. This study investigated pyr-amiding genes resistant to blast, BB and BPH to develop restorer lines. Ten new lines with blast, BB and/or BPH resistance genes were developed using marker-assisted selection （MAS） technique and agronomic trait selection （ATS） method. Only HR13 with resistance genes to blast, BB and BPH was obtained. In addition to blast and BB resistance, four lines （HR39, HR41, HR42, HR43） demonstrated moderate resistance to BPH, but MAS for BPH resistance genes were not conducted in developing these four lines. These data suggested that there were unknown elite BPH resistance genes in the Zhongzu 14 donor parent. A more effective defense was demonstrated in the lines withPi1 andPi2 genes although the weather in 2012 was favorable to disease incidence. Blast resistance of the lines with a single resistance gene,Pita, was easily inlfuenced by the weather. Overal, the information obtained through pyramiding multiple resistance genes on developing the restorer lines is helpful for rice resistance breeding.

Effects of Temperature on Functional Response of Anagrus nilaparvatae Pang et Wang (Hymenoptera:Mymaridae) on the Eggs of Whitebacked Planthopper,Sogatella furcifera Horvth and Brown Planthopper,Nilaparvata lugens Stl

Understanding the temperature affecting parasitic efficiency is critical to succeed in utilizing parasitoid as natural enemy in pest management. Laboratory studies were carried out to determine the effects of temperature on parasitoid preference of female Anagrus nilaparvatae Pang et Wang （Hymenoptera：Mymaridae） to the eggs of whitebacked planthopper （WBPH）, Sogatella furcifera Horváth and brown planthopper （BPH）, Nilaparvata lugens Stl to build a composite model describing changes in parasitic response along a temperature gradient （18, 22, 26, 30, 34°C）. The results showed that attack responses of A. nilaparvatae on WBPH and BPH were the best described by a Type II functional response. The two parameters, attack rates （a） and handling times （Th）, of A. nilaparvatae to both eggs were influenced by the temperature. The maximum attack rates to WBPH （1.235） and BPH （1.049） were at 26 and 34°C, respectively, and the shortest handling times to WBPH （0.063） and BPH （0.057） were at 30 and 26°C, respectively. However, the optimal temperature for parasitic efficiency of A. nilaparvatae to WBPH and BPH eggs was both at 26°C, which showed that the present microclimate temperature of the habitat in the paddyfield was beneficial to A. nilaparvatae and indicated that parasitic efficiency of A. nilaparvatae would be impaired by global warming.