Nutrient acquisition by insect herbivores affect all aspect of the the lifespan of individauls. For seed-sucking insect herbivores, they face challenges with nutrient acquisition due to requirement for extra-oral digestion of seed contents into a readily-ingestible state. In this study, we demonstrated environmentally-transmitted Caballeronia insecticola allow seed-sucking R. pedestris to overcome challenges with extra-oral digestion. Through the evaluation, first, we found symbiotic insects exhibited enhanced feeding efficiency by consuming significantly larger amount of food per feeding attempt compared to apo-symbiotic insects (P<0.05). Then, we observed feeding behavior modification in the symbiotic insects from the behavior tracking evaluation. Symbiotic insects displayed dichotomic behavior which can be generally divided into early focused feeding and later subdued resting periods. By contrast, apo-symbiotic insects displayed unordered behavior by frequent switches between feeding and walking behavior.

The bean bug, Riptortus pedestris, is one of major hemipteran pests against the soybean, Glycine max in Korea. In previous study, it was suggested that water-taking in the first instar nymphal stage of R. pedestris is certainly necessary for successful ecdysis and their survival. To investigate water-taking behavior of the first instar nymphs of R. pedestris on plant, electrical penetration graphic patterns were analysed while R. pedestris fed on the napa cabbage leaf, Brassica rapa subsp. pekinensis by using EPG system. In the result, a X-waveform was dominantly observed for average 42.0 min out of total recording time (120 min). There was significant positive correlation (r = 0.457, P = 0.000452, n = 55) between body weight increase and cumulative durations of X-waveform. In addition, histological observation of cross-sections indicated that stylet tip in the plant tissue during X-waveform period is located nearby xylem cells, suggesting that X-waveform is closely related to xylem sap feeding behavior of R. pedestris in plant.

The bean bug is one of major hemipteran pests against the soybean, Glycine max in Korea. Gene expression regulation technologies such as RNA interference and gene editing have emerged as a next generation pest control strategy in agriculture. To search genes effective to feeding inhibition with exogenous dsRNA, a total of 26,761 transcripts were obtained from salivary glands of the bean bug and several target gene candidates such as GPCR, OBP, and CSP were profiled in this study. On the other hand, 1st instar nymphs of the bean bug after hatching were highly attracted to water and absorbed about 701.5 ± 137.72 ㎍/individual/day, suggesting that it could be a practical way for RNA interference by feeding exogenous dsRNA dissolved in water. These preliminary results are expected to be helpful to investigate novel pest control strategy of R. pedestris in the future.

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

A novel insect-infecting positive sense single-stranded RNA virus, Riptortus pedestris virus-1 (RiPV-1), was found in the Riptortus pedestris transcriptome data by de novo assembly and further confirmed by RACE method. The genome of RiPV-1 consists of 10,554 nucleotides (nt) excluding the poly(A) tail and contains a single large open reading frame (ORF) of 10,371 nt encoding a 3,456 aa polyprotein and flanked by 71 and 112 nt 5' and 3' noncoding regions, respectively. RiPV-1 genome contains the consensus genome organization of picorna-like RNA helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp) array in that order from the 5' to the 3' end. From the phylogenetic analysis, RiPV-1 was clustered with unassigned insect RNA viruses, APV and KFV, which suggests that these three insect picorna-like viruses might constitute a novel group of insect-infecting RNA viruses. Tissue tropism analysis revealed that RiPV-1 was relatively abundant in the thorax, abdomen, midgut and fat body. Interestingly, RiPV-1 replication was enhanced by Beauveria bassiana JEF-007 infection that was quantified using qRT-PCR. This study identified a novel insect-infecting virus and provided further insight into the relationship between virus, fungus and host.

Beauveria bassiana (Bb) is an entomopathogenic fungus with a wide host range, and is commonly used as an environment-friendly biopesticide. However, the molecular mechanisms of Bb-host interactions are not well understood. Here, RNA isolated from a highly virulent strain of B. bassiana (Bb JEF-007) and Riptortus pedestris (Hemiptera: Alydidae) (bean bug) infected with this strain were subjected to high throughput next generation sequencing (NGS) to analyze and compare transcriptomes. Differentially expressed gene (DEG) analysis showed that 2,381 genes were up-regulated and 2,303 genes were down-regulated upon infection. Most DEGs were classified into the categories of single-organism, cellular and metabolism processes by gene ontology (GO) analysis. Carbon metabolism-related enzymes in the glyoxylate cycle were significantly up-regulated, suggesting a possible role for them in Bb growth in the host. This work provides insight into how entomopathogenic B. bassiana occupies agriculturally harmful bean bug at the late stage, which might be essential during fungal infection.

Bean bug, Riptortus pedestris (Hemiptera: Alydidae) is an agriculturally serious pest in East Asian countries. Chemical pesticides have been contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Herein entomopathogenic Beauveria bassiana JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by contact-exposure and spray methods. The Agrobacterium tumefaciens-mediated transformation of B. bassiana JEF-007 generated random transformants and some mutants showed reduced virulence against Tenebrio molitor (Coleoptera: Tenebrionidae) larvae and R. pedestris nymph. Compared to the wild-type, the two transformants showed remarkably different morphology, conidial production, and thermotolerance. To figure out pathogenicity-related genes, thermal asymmetric interlaced (TAIL) PCR of the random transformants was performed and possibly some virulence-related genes were predicted. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.

Although expression of foreign genes in a crop species has been target techniques for powerful protection against insect pests, classical breeding programs using varietal resistance of a crop are still being processed. It is, however, frequently difficult to find key products expressed from resistance-related genes in the variety, and those action mechanism. Here, an unterminated story about seed of a mungbean (Vigna radiata) variety with high resistance against the adzuki bean weevil and the bean bug is introduced. The two insects cannot survive on seeds of the variety, although they well develop on susceptible ones. Molecular markers linked to the resistance were selected through BAC library screening and near isogenic lines, and finally a bruchid resistance gene was suggested after map-based cloning approaches. Starvation and chemicals were suggested for a resistant mechanism and a related factor through feeding and behavior experiments, respectively. The seed flour was extracted with organic solvents, and isolated into several fractions on chromatography. Several peaks on HPLC in a fraction were related with the high mortality of the bean bug. A partial structure in an isolated chemical was observed before full identification.

Bean bug, Riptortus pedestris is an agriculturally serious pest in East Asian countries, reducing the value of crop quality and loss of income in agribusiness. Chemical pesticides have contributed to the management of the pest, but nowadays insect resistance limits the use of chemical pesticides, thus alternatively new pesticides with different mode of actions such as entomopathogenic fungi are considered. Beauveria bassiana and Metarhizium anisopliae JEF isolates were collected, identified and assayed against bean bugs in laboratory conditions. Some isolates showed >80% virulence by spray and contact-exposure methods. Supernatant showed different level of enzyme activity including chitinase, Pr1 protease and lipase. The Agrobacterium tumefaciens-mediated transformation generated random transformants and some mutants had reduced virulence. TAIL-PCR of the random transformants revealed virulence-related genes. This work can be a strong platform for the functional genetics of bean bug-pathogenic B. bassiana.