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.

Analysis of Resistance-related Proteins in Rice Against Brown Planthopper by Two-dimensional Electrophoresis

A recombinant inbred population (RI) was constructed from a cross between B5, an introgression. line from the wild rice Oryza officinalis Wall. ex Watt, and susceptible cultivar Minghui 63 ( O. sativa L.). The brown planthopper ( BPH) resistances of RI lines were evaluated. Based on bulked segregant analysis (BSA), two protein bulks were made by extracting proteins from equally mixed seedlings of extremely resistant and susceptible plants selected from the RI population, respectively. Two-dimensional electrophoresis was used to detect the changes of polypeptide pattern. Results showed that a protein P40 ( pI 6.3, Mw 40 kD) was significantly reduced or vanished after BPH infestation for 48 h in the susceptible bulk, while it remained uninfluenced in the resistant bulk. In connection with the physiological changes of the resistant and susceptible lines subjected to BPH sucking, we suppose that the protein P40 is related to the interaction responses of lice plants to BPH infestation.

Relationship between Rice Planthopper Occurrence Area in China and Atmospheric Circulation Indices

[Objective] This study aimed to establish models based on atmospheric cir- culation indices for forecasting the area attacked by rice planthopper every year, and to provide guide for preventing and controlling planthopper damage. [Method] The data related to rice planthopper occurrence and atmospheric circulation were collected and analyzed with the method of stepwise regression to establish the prediction models. [Result] The factors significantly related to the area attacked by rice plan-thopper were selected. Two types of prediction models were established. One was for Sogatella furcifera （Horvath）, based on Atlantic-Europe circulation pattern W in October in that year, Pacific polar vortex area index in October in that year, North America subtropical high index in August in that year, Atlantic-Europe circulation pattern W in June in that year, northern boundary of North America subtropical high in February in that year, Atlantic-Europe polar vortex intensity index in October in that year and Asia polar vortex intensity index in November in the last year; the other type of prediction models were for Nilaparvata lugens （Stal）, based on the Eastern Pacific subtropical high intensity index in July in that year, northern hemi- sphere polar vortex area index in October in the last year, Asia polar vortex strength index in November in the last year, north boundary of North America-At- lantic subtropical high in September in that year, north boundary of North Africa-At- lantic-North America subtropical high in January in that year, sunspot in September of the last year and eastern Pacific subtropical high area index in September in that year. [Conclusion] With the stepwise regression, the forecasting equations of the rice planthopper occurrence established based on the atmospheric circulation indices could be used for actual forecast.

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.

Spectrum Detection of Rice Planthopper Populations from Canopy Reflectance Based on Principal Component Regression

[ 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.

Study on the Meteorological Condition Grade Forecast for the Migration of Rice Planthopper

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.

Density Restriction Effects of Reproductive Rate of Rice Brown Planthopper Population in Super Rice Yongyou 6

[ 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.

The Control Effect of Prevention and Control Technology Combination against Southern Rice Black-Streaked Dwarf Disease

Objective] This study aimed to explore a better prevention and control technology combination against southern rice black-streaked dwarf disease. [Method] The control effects of seed dressing, antiviral agents and fly net along with 25%pymetrozine against southern rice black streaked dwarf disease were determined. [Result] When the seeds were dressed with 60% imidacloprid （FS, 2 g a.i./kg）, and the rice seedlings were sprayed once with 25% pymetrozine （WP, 360 g/hm2） 10 d before the transplanting and sprayed twice with 30% Dufulin （WP, 800×） at three-leaf stage and 7 d after the transplanting respectively, the control efficiency reached 88.05%. When fly net and pesticides were applied simultaneously, the fly net was lifted and 25% pymetrozine （WP） was spayed once on the rice seedlings at six-leaf stage and the seedlings were hardened for 3 d and sprayed once with 25%pymetrozine （WP） 10 d after the transplanting, the control efficiency reached 80.50%. [Conclusion] Seed dressing or applying antiviral agents alone can not better control diseases. The southern rice black-streaked dwarf disease can be better con-trol ed by seed dressing, along with spraying of planthopper-kil ing agents at two-leaf stage, 3 d before the transplanting and 7 d after the transplanting, respectively. If condition al ow, fly net can be instal ed to achieve better control effect.