The vegetative insecticidal protein Vip3 produced by Bacillus thuringiensis is a tetramer composed of five domains. Its N-terminal is mainly related to the stability of the structure, while the C-terminal is a potential specific receptor binding region. The Vip3 family of vegetative insecticidal proteins contains 14 holotypes and more than 110 proteins. The proposed mode of action of Vip3 shares some similarities with that of the crystal proteins (Cry proteins), in that both undergo activation (proteolytic processing) in the insect midgut, bind to receptors on the surface of the midgut cells, make pores that lead to cell lysis, and eventually death of the insect. In consideration of different active mechanisms to insects as well as their insecticidal spectrum complementarity, the stack strategy of two kinds of genes (vip3 and cry) has been widely used in development of new insect-resistant transgenic crops worldwide; and Vip3Aa transgenic crops such as maize and cotton have been commercially planted in the United States, Brazil and other countries, in order to delay and manage the Cry protein resistance from some target pests such as Spodoptera frugiperda and others. It has been reported that a high resistance to Vip3 toxins by some insects could be rapidly evolved under the toxin selective press in the laboratory, and also field monitoring to several species has confirmed resistance occurrence of Vip3 in natural environments. Therefore, monitoring and management of target pest resistance are much necessary for industrialization of Vip3 insect-resistant transgenic crops.

RNA interference (RNAi) is a highly conserved gene-silencing phenomenon, induced by 20-30 nucleotide (nt) small molecules of non-coding RNA in vivo. Three non-coding RNAs, such as small interfering RNA (siRNA), microRNA (miRNA) and PIWI-interacting RNA (piRNA), play important roles in RNAi of eukaryotes including insects. The applications of RNAi in pest control are mainly on the research of gene function, transgenic insect-resistant plants and novel nucleic acid pesticides. Before the applications, the safety risks in terms of non-target effects, target pest resistance and environmental persistence of interfering RNAs need to be strictly evaluated. In this paper, the mechanism of RNAi technology, application and safety of RNAi in agro-forestry pest control are described, which provide some theoretical basis for RNAi technology in pest control.

Rice planthoppers, mainly including Nilaparvata lugens, Sogatella furcifera, and Laodelphax striatellus, are one of the most important insect pests of rice in China. In this review, we summarized the latest research progress on the molecular basis of important genetic characteristics (wing-morph differentiation, fecundity, insecticide resistance) of rice planthoppers, interactions among rice, rice planthoppers, natural enemies of rice planthoppers and other organisms, mechanisms underlying rice planthopper outbreak, and management of rice planthoppers. Finally, we suggest that future studies should further dissect the molecular basis of biology and ecology related to rice planthopper outbreak, and find the coordination mechanisms between intensified agriculture and rice ecosystem resistance at the micro level, so as to maintain or improve the rice ecosystem resistance, and achieve sustainable management of rice planthoppers in the context of intensified agriculture.

Insects are capable of inducing the production of heat shock proteins when exposed to environmental stresses or under specific physiological conditions such as diapause. The transcriptional activation of heat shock proteins is usually controlled by the heat shock transcription factor (HSF), a class of transcription factors that are widely present in the organism. There are numerous HSF genes in plants and mammalian species, while previous studies suggested that many insects only had single HSF gene. Under normal conditions, HSF proteins are often present in the inactive form in cells, and they are activated upon shock, and then transferred to the nucleus, where they bind to specific functional domains of target genes and eventually activate the transcription of target genes to improve the tolerance of the whole organism. This paper reviewed the research progress on the basic structure, transcriptional regulation, and biological function of HSF in insects to deepen the understanding of the regulatory role of HSF in stress response mechanisms of insects, and provide new ideas for agroforestry pest control.

In recent years, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (CRISPR/Cas) system-mediated gene editing technology has become an important research tool in modern plant molecular biology and agricultural breeding due to its simplicity, high efficiency, generality and accuracy. Plant virus disease seriously endangers the normal growth and development of plants, and causes devastating damages to crop production worldwide. CRISPR/Cas gene editing technology can effectively target DNA virus and RNA virus sequences, and improve the resistance of host plants to viruses. In addition, gene editing technology can edit plant endogenous genes which are conducive to viral infection or replication to create new antiviral germplasm resources. This study reviewed the CRISPR/Cas gene editing technology and elaborated the research progress and examples of using this system to achieve antiviral infection. The potential application prospects and limitations of virus-induced gene editing system were also discussed. Finally, the advantages and challenges of CRISPR/Cas gene editing technology in plant antiviral research were pointed out, which will be useful for guiding innovation of germplasms for virus resistance.

Rice blast caused by Magnaporthe oryzae is a worldwide disease, which seriously affects the global rice production. The rapid variation of M. oryzae resulted in a decline in resistant variety screening and pesticide control. Therefore, revealing the pathogenic mechanism of M. oryzae at the molecular level can provide target molecules for rice disease control and new fungicide development. The utilization of endophytic fungi from wild rice to control rice blast provides a new idea for green management. By excavating the resources of wild rice endophytic fungi, identifying functional strains with the activities of promoting growth, immune induction and resistance to adverse conditions, and exploring the interaction mechanism between rice and endophytic fungi, it provides a basic theory for the development of endophytic fungi as new biocontrol agents. This review mainly summarized the novel research results on the pathogenic mechanism of M. oryzae and the functional mechanism of endophytic fungi in rice in recent years, and look forward to the future development direction of a new green prevention and control strategy for M. oryzae.

Fire blight is an important disease caused by Gram-negative bacterium Erwinia amylovora that severely harms Rosaceae plants, and it is an important factor to restrict the development of pear and apple industries in China. Erwinia amylovora is highly virulent with a wide host range, and thus it is difficult to find a unified target for its eradication. With the emergence of drug-resistant strains, control of the disease becomes more difficult. In this paper, the disease characteristics, epidemiology, management methods and the mechanisms underlying E. amylovora pathogenicity and plant resistance were reviewed, which could provide references to further research and efficient control of fire blight.

negevirus (nege-like virus) is a newly proposed virus taxon of insect-specific RNA viruses and has been found in various insect species. In this study, the complete genome of a novel nege-like virus, named as “Nbu Aulacophora lewisii nege-like virus 1” (NbuALNV-1), was identified from a single leaf beetle, Aulacophora lewisii. NbuALNV-1 is a positive-sense single-stranded RNA virus with the genome of 9 832 nucleotide (nt) in length (excluding the polyA tail), with a 5′ untranslated region (UTR) of 292 nt and a 3′ UTR of 77 nt, and six open reading frames (ORFs). Five conserved domains of negeviruses were predicted to be encoded by the genome of NbuALNV-1, consisting of the viral methyltransferase domain, RNA virus helicase domain, and RNA-dependent RNA polymerase (RdRp) domain in ORF1, and the DISB-ORF2_chro domain and SP24 domain in the other two ORFs. Phylogenetic analysis of RdRp amino acid sequences of NbuALNV-1, together with Hubei Wuhan insect virus 9, Aphis glycines virus 3, and Fort Crockett virus showed that they formed as an “unclassified” clade and closely related to members in plant viruses of the family Kitaviridae. Furthermore, typical characteristics of virus-derived small interfering RNAs were observed by NbuALNV-1-derived small interfering RNAs, indicating that NbuALNV-1 might induce the host RNA interfering antiviral immune response. In summary, a nege-like virus was identified in A. lewisii, and NbuALNV-1 is the first negevirus discovered in the insect order Coleoptera.

The whitefly Bemisia tabaci, an important agricultural insect pest, secretes saliva into plants while feeding on them. Our previous study found that the whitefly secretes the salivary effector protein BtArmet (Bemisia tabaci arginine rich, mutated in early stage of tumors) to inhibit plant resistance by targeting tobacco cystatin. In this study, we continued to screen tobacco proteins that interact with the salivary effector protein BtArmet. Yeast two-hybrid and bimolecular fluorescence complementation (BiFC) assays showed that Nicotiana tabacum WRKY51 interacted with BtArmet. The up-regulation of NtWRKY51 gene in tobacco was significantly induced by whitefly infestation. However, virus-induced gene silencing (VIGS) assays combined with bioassay of whiteflies showed that the fecundity of NtWRKY51-silenced tobacco plants by female whiteflies was significantly lower than that of the negative control (tobacco with silenced empty vector). Silencing NtWRKY51 gene had no effect on phytohormone signaling pathways mediated by salicylic acid or jasmonic acid in tobacco plants. These results provide references for further investigations on the molecular mechanisms underlying plant resistance to whiteflies.

Siniphanerotomella He, Chen et van Achterberg, 1994 is an endemic genus to China with only two species currently known worldwide. A third species from Guangdong Province in China, S. chengjiaani Tang et Chen, sp. nov., is described and illustrated. A key to all known species of the genus is also provided in this study.

Neuropeptides are crucial for life activities and environmental fitness of insects, which are potential targets for pest control. The brown planthopper (Nilaparvata lugens) is an important pest on rice in Asia. Thus, functional screening analysis of genes encoding neuropeptides and their receptors in N. lugens were conducted. The cDNA sequences of 41 neuropeptides (containing one alternative splicing transcript) and 44 receptors were verified by polymerase chain reaction (PCR). Four neuropeptide genes (NlCCAP, NlETH, NlOKA and NlPK) and two receptor genes (NlA36 and NlA46) were identified to cause high mortality through RNA interference (RNAi), showing potential in pest control. These genes knockdown resulted in the relative survival rate of adult less than 0.2 when injected with double-stranded RNA (dsRNA) at the third instar, and had the potential to be target genes for pest control. The results of RNAi also demonstrated that neuropeptides, such as crustacean cardioactive peptide (CCAP), ecdysis triggering hormone (ETH) and bursicon were conserved in physiological function, while other neuropeptides (eg. prothoracicotropic hormone, eclosion hormone) which related to ecdysis in other insects were undefined in physiological function in N. lugens. Our study provides a basis for further exploration of neuropeptides’ physiological function in N. lugens.

In order to establish an accurate detection method for royal jelly freshness, the major royal jelly proteins (MRJPs) in royal jelly by trypsin enzymatic hydrolysis were identified by ultra-high performance liquid chromatography-tandem quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF/MS). The primary sequence data of eight MRJPs (MRJP1-7 and MRJP9) were identified by proteomics analysis platform. Then three specific marker polypeptides were obtained according to the criteria of highly easy enzymolysis and high stability, including YNGVPSSLNVISK (MRJP1), TLQMIAGMK (MRJP2), and LTVAGESFTVK (MRJP3). The three specific marker polypeptides of MRJP1-3 and their isotopic marker peptides were synthesized, respectively, and an ultra-high performance liquid chromatography-triple quadrupole tandem mass spectrometry (UPLC-TQMS) method for the quantitative detection of MRJP1-3 in royal jelly was established. The results showed that the MRJP1 polypeptide had a good linear relationship in the range of 10-1 600 ng/mL, and MRJP2 and MRJP3 polypeptides had a good linear relationship in the range of 10-600 ng/mL, and the correlations of determination (R²) were all more than 0.99. Finally, by analyzing the degradation degrees of MRJP1-3 in royal jelly with the aging time, it was confirmed that the degradation rates of MRJP1-3 were all positively correlated with the aging time at 40 ℃ (R²＞0.9). It is indicated that the MRJP1-3 detection method established in this study is highly specific, which provides technical support for the quality evaluation of royal jelly.

To explore the courtship, mating and oviposition behavior of Chilo suppressalis (Walker) in a high dose sex pheromone environment, the mating disruption was simulated in a wind tunnel, and the sex pheromone titers were analyzed in combination with behavioral observation and chemical methods. The results showed that in the wind tunnel, the high dose of Z11-16:Ald and the ternary blend [m(Z11-16:Ald)∶ m(Z9-16:Ald)∶m(Z13-18:Ald)=10∶1∶1.2] of 3.1 mg per spraying significantly inhibited the mating rates and sex pheromone titers of female moths, but no significant differences were found between the two treatments. There were no significant differences in the number of mating pairs of male and female moths and the time interval between each spraying. The mating rates of male moths with the removal of antennae decreased to (8.3±1.1)% and (10.0±8.9)%, respectively, in the environment of spraying or not spraying high dose Z11-16:Ald. The number of laid eggs and egg hatching rates were not affected in the high dose sex pheromone environment. The ages of female moths significantly affected the mating rates with male moths. The mating rates of 1, 3 and 5 day-old female moths with 1 day-old male moths were (60.0±5.2)%, (31.7±6.0)%, and (8.3±3.1)%, respectively. The number of laid eggs and egg hatching rates of female moths decreased with the ages. In conclusion, the high dose sex pheromone Z11-16:Ald and complete blend can inhibit the mating rates and sex pheromone titers of C. suppressalis, and have no impact on the number of laid eggs and egg hatching rates of female moths, but the ages of female moths can affect the number of laid eggs and egg hatching rates; and the decrease in mating rates might be related to the inactivation of olfactory system of male moths.

The brown planthopper (Nilaparvata lugens) is an important rice pest. In this study, the tomographic images of adult brown planthopper were obtained by using micro-computer tomography technology (Micro CT). Three-dimensional models of internal tissues and organs of the brown planthopper were established by combining manual modeling and deep learning. Three-dimensional models of the central nervous system, muscle tissue, alimentary canal and reproductive system of the brown planthopper were obtained. These models preserved the original morphologies of these structures and accurately restored the spatial arrangement of all tissues and organs within the brown planthopper. Measurements of the internal organization of insects were analyzed using Dragonfly software. This study established and refined the three-dimensional reconstruction technique of insect tissues and organs, which can contribute to a more precise view of the internal organization structure of insects and can be used for phenotypic observation of insect morphology and organ development. It provides a new technique for insect development investigation and gene function analysis in entomology.

The intraguild predation (IGP) level among the main generalist natural enemies is related to their population occurrence and the effectiveness of “green control” strategies for insect pests in agroecosystem. In this study, the detection systems of quantitative real-time polymerase chain reaction (qPCR) probe were first established and then optimized for analyzing the DNA of Pardosa pseudoannulata and Paederus fuscipes in ratoon rice fields. Then the detection systems were used to analyze the IGP levels between the two predators in ratoon rice fields. On this basis, the IGP and their biological and abiotic factors affecting the IGP level were studied under indoor conditions. The results showed that the primers and probes designed for the two predators had strong specificity. The established and optimized qPCR detection systems had good amplification repeatability and high sensitivity for the target genes of the two predators. After the system was used to detect 1 527 individuals of the two predators from ratoon rice fields and the indoor IGP tests, it was found that there was a ubiquitous and strong bidirectional IGP between the two predators in the rice field. The role transformation of intraguild predator or prey in the IGP and the influence of ambient temperature on the IGP level were closely related to their relative body sizes (developmental stages). The IGP level can be affected through extraguild preys and abiotic factors and their combination to various extents. These results enrich the IGP theory among arthropod predators in the rice ecosystem, and lay good foundations for further IGP study of natural enemies.

As an important quarantine pest in the world, it is extremely crucial to identify some mosquitoes at ports in China through online image comparisons. The typical morphometric characters of head, thorax and abdomen of adult female mosquitoes were divided into multiple anatomical features under the frame of the current taxonomic system in China, and these features were further split into two sub-selections in the dichotomous parallel mode, and then these features and sub-selections were linked with relative entities of mosquito genera by the internationally popular insect classification diagnostic software Lucid Professional v3.5. Finally, a latest online key to mosquito taxonomy of China (Diptera: Culicidae) was generated by a classification and diagnosis network service system Lucid Key Server v1.0. This key was based on a matrix of 34 candidate features, 68 alternative sub-selections, 89 precise illustrations of the representatives of each genus, along with the quality database of mosquito genera created by a biodiversity automatic database building software Fact Sheet Fusion v1.0. The users could distinguish the unknown specimen from 2 subfamilies Anophelinae and Culicinae, 10 tribes, 21 genera (including 2 subgenera) by clicking on any of the alternative features or sub-selections. To sum up, this interactive multiplex intelligence identification system to mosquito genera has good image comparison and recognition functions, and can provide an effective tool in the morphological identification of important vectors of mosquito-borne diseases.

The Chinese banana industry is under grave threat by Fusarium wilt of banana caused by Fusarium oxysporum f. sp. cubense (Foc). However, it is still obscure how to control this devastating disease effectively. In this study, the fungicidal activities of four triazole fungicides were tested by hyphae growth rates of Foc tropical race 4 (Foc TR4). The results showed that the fungicidal activities of these four fungicides were prothioconazole＞tebuconazole＞propiconazole＞myclobutanil. The morphology changes and membrane integrity of Foc TR4 in response to these four fungicides were evaluated by morphological observation of hyphae, and detection of relative conductivity and malondialdehyde (MDA) concentrations. Compared with the control group, all these four fungicides can cause abnormalities including increased hyphae branching, irregular distorted, shrivelled, sunken and flattened in hypha cells of Foc TR4. The MDA concentrations and membrane permeability of hypha cells were significantly elevated in all treatment groups. The activities of the cytochrome P450 enzymes were increased significantly so were the expression levels of cytochrome P450 sterol 14α-demethylase (CYP51) genes CYP51-1 and CYP51-3 upon treatment with these fungicides. Molecular docking and surface plasmon resonance (SPR) assays were employed to demonstrate interaction modes of four fungicides with CYP51 of Foc TR4. Even though prothioconazole showed less binding affinity with CYP51 than other three fungicides, it showed the strongest fungicidal activity against Foc TR4, which indicated the specificity of its mode of interaction with CYP51. This study can provide some theoretical references for the screening and rational design of novel fungicides for controlling Fusarium wilt of banana pathogen of Foc.