Plants undergo several but very precise molecular, physiological, and biochemical modulations in response to biotic stresses. Mitogen-activated protein kinase (MAPK) cascades orchestrate multiple cellular processes including plant growth and development as well as plant responses against abiotic and biotic stresses. However, the role of MAPK kinases (MAPKKs/MKKs/MEKs) in the regulation of plant resistance to herbivores has not been extensively investigated. Here, we cloned a rice MKK gene, OsMKK3, and investigated its function. It was observed that mechanical wounding, infestation of brown planthopper (BPH) Nilaparvata lugens, and treatment with methyl jasmonate (MeJA) or salicylic acid (SA) could induce the expression of OsMKK3. The over-expression of OsMKK3 (oe-MKK3) increased levels of jasmonic acid (JA), jasmonoyl-L-isoleucine (JA-Ile), and abscisic acid (ABA), and decreased SA levels in rice after BPH attack. Additionally, the preference for feeding and oviposition, the hatching rate of BPH eggs, and BPH nymph survival rate were significantly compromised due to over-expression of OsMKK3. Besides, oe-MKK3 also augmented chlorophyll content but impaired plant growth. We confirm that MKK3 plays a pivotal role in the signaling pathway. It is proposed that OsMKK3 mediated positive regulation of rice resistance to BPH by means of herbivory-induced phytohormone dynamics.

Gibberellins (GAs) play pivotal roles in plant growth and development, and in defenses against pathogens. Thus far, how the GA-mediated signaling pathway regulates plant defenses against herbivores remains largely unknown. In this study, we cloned the rice GA receptor gene OsGID1, whose expression was induced by damage from the brown planthopper (BPH) Niaparvata lugens, mechanical wounding, and treatment with salicylic acid (SA), but not jasmonic acid. The overexpression of OsGID1 (oe-GID1) decreased BPH-induced levels of SA, H₂O₂, and three SA-pathway-related WRKY transcripts, but enhanced BPH-induced levels of ethylene. Bioassays in the laboratory revealed that gravid BPH females preferred to feed and lay eggs on wild type (WT) plants than on oe-GID1 plants. Moreover, the hatching rate of BPH eggs on oe-GID1 plants was significantly lower than that on WT plants. In the field, population densities of BPH adults and nymphs were consistently and significantly lower on oe-OsGID1 plants than on WT plants. The increased resistance in oe-GID1 plants was probably due to the increased lignin level mediated by the GA pathway, and to the decrease in the expression of the three WRKY genes. Our findings illustrated that the OsGID1-mediated GA pathway plays a positive role in mediating the resistance of rice to BPH.

The brown planthopper (BPH) impacts both rice yield and quality. The exogenous application of abscisic acid (ABA) and jasmonic acid (JA) has been previously shown to induce rice resistance to BPH; however, the regulation of rice-mediated defense by these plant growth regulators is unclear. We applied exogenous JA and ABA to rice and analyzed molecular responses to BPH infestation. Nine RNA libraries were sequenced, and 6218 differentially expressed genes (DEGs) were generated and annotated. After ABA + BPH and JA + BPH treatments, 3491 and 2727 DEGs, respectively, were identified when compared with the control (BPH alone). GO enrichment and KEGG pathway analysis showed that the expression of several JA pathway genes (OsAOS2, encoding allene oxide synthase; OsOPR, 12-oxo-phytodienoic acid reductase; and OsACOX, acy1-CoA oxidase) were significantly up-regulated after ABA + BPH treatment. Furthermore, exogenous JA increased the expression of genes involved in ABA synthesis. Meanwhile, the expression levels of genes encoding WRKY transcription factors, myelocytomatosis protein 2 (MYC2) and basic leucine zippers (bZIPs) were up-regulated significantly, indicating that ABA and JA might function together to increase the expression of transcription factors during the rice defense response. The DEGs identified in this study provide vital insights into the synergism between ABA and JA and further contribute to the mechanistic basis of rice resistance to BPH.

The oriental armyworm, Mythimna separata (Lepidoptera: Noctuidae) is a harmful pest which causes heavy loss in agricultural crops. Sex identification of M. separata is very important for understanding the biology and management of this pest. Development of fast and convenient methods to identify M. separata female and male is the necessary prerequisite for conducting gender-related studies. In this study, new methods to differentiate sex of M. separata at pupal and adult stages were investigated. Our results showed that posterior abdominal segments containing genital organs which are morphological traits to differentiate female and male. Genital openings are important characters for the female and male pupae differentiation. For female pupa, it locates on the ventral side of eighth and ninth abdominal segments and appears as a visible longitudinal suture; for male pupa, it locates on the ventral side of the ninth abdominal segment and appears as a short slit surrounded by a pair of visible semicircular lumps. Bristle numbers are another indicator for sex differentiation of adults. Each female has three-bristled frenulum and each male has one bristled-frenulum near the base of hind wing of each side of moth body. We confirmed the two sex differentiation methods manually by dissecting genitalia of adults and found their genger was as same as what we examined the sex of pupae and adults using the morphological methods.

The pentose phosphate pathway (PPP) plays an important role in plant growth and development, and plant responses to biotic and abiotic stresses. Yet, whether the PPP regulates plant defenses against herbivorous insects remains unclear. In this study, we cloned a rice cytosolic 6-phosphogluconate dehydrogenase gene, Os6PGDH1, which encodes the key enzyme catalyzing the third step in the reaction involving the oxidative phase of the PPP, and explored its role in rice defenses induced by brown planthopper (BPH) Nilaparvata lugens. Levels of Os6PGDH1 transcripts were detected in all five examined tissues, with the highest in outer leaf sheaths and lowest in inner leaf sheaths. Os6PGDH1 expression was strongly induced by mechanical wounding, infestation of gravid BPH females, and jasmonic acid (JA) treatment. Overexpressing Os6PGDH1 (oe6PGDH) decreased the height of rice plants and the mass of the aboveground part of plants, but slightly increased the length of plant roots. In addition, the overexpression of Os6PGDH1 enhanced levels of BPH-induced JA, jasmonoyl-isoleucine (JA-Ile), and H₂O₂, but decreased BPH-induced levels of ethylene. Bioassays revealed that gravid BPH females preferred to feed and lay eggs on wild-type (WT) plants over oe6PGDH plants; moreover, the hatching rate of BPH eggs raised on oe6PGDH plants and the fecundity of BPH females fed on these were significantly lower than the eggs and the females raised and fed on WT plants. Taken together, these results indicate that Os6PGDH1 plays a pivotal role not only in rice growth but also in the resistance of rice to BPH by modulating JA, ethylene, and H₂O₂ pathways.