Silencing an ATP-Dependent Caseinolytic Protease Proteolytic Subunit Gene Enhances the Resistance of Rice to Nilaparvata lugens.

The ATP-dependent caseinolytic protease (Clp) system has been reported to play an important role in plant growth, development, and defense against pathogens. However, whether the Clp system is involved in plant defense against herbivores remains largely unclear. We explore the role of the Clp system in rice defenses against brown planthopper (BPH) Nilaparvata lugens by combining chemical analysis, transcriptome, and molecular analyses, as well as insect bioassays. We found the expression of a rice Clp proteolytic subunit gene, OsClpP6, was suppressed by infestation of BPH gravid females and mechanical wounding. Silencing OsClpP6 enhanced the level of BPH-induced jasmonic acid (JA), JA-isoleucine (JA-Ile), and ABA, which in turn promoted the production of BPH-elicited rice volatiles and increased the resistance of rice to BPH. Field trials showed that silencing OsClpP6 decreased the population densities of BPH and WBPH. We also observed that silencing OsClpP6 decreased chlorophyll content in rice leaves at early developmental stages and impaired rice root growth and seed setting rate. These findings demonstrate that an OsClpP6-mediated Clp system in rice was involved in plant growth-defense trade-offs by affecting the biosynthesis of defense-related signaling molecules in chloroplasts. Moreover, rice plants, after recognizing BPH infestation, can enhance rice resistance to BPH by decreasing the Clp system activity. The work might provide a new way to breed rice varieties that are resistant to herbivores.

Silencing a Simple Extracellular Leucine-Rich Repeat Gene OsI-BAK1 Enhances the Resistance of Rice to Brown Planthopper Nilaparvata lugens.

Many plant proteins with extracellular leucine-rich repeat (eLRR) domains play an important role in plant immunity. However, the role of one class of eLRR plant proteins-the simple eLRR proteins-in plant defenses against herbivores remains largely unknown. Here, we found that a simple eLRR protein OsI-BAK1 in rice localizes to the plasma membrane. Its expression was induced by mechanical wounding, the infestation of gravid females of brown planthopper (BPH) Nilaparvata lugens or white-backed planthopper Sogatella furcifera and treatment with methyl jasmonate or abscisic acid. Silencing OsI-BAK1 (ir-ibak1) in rice enhanced the BPH-induced transcript levels of three defense-related WRKY genes (OsWRKY24, OsWRKY53 and OsWRKY70) but decreased the induced levels of ethylene. Bioassays revealed that the hatching rate was significantly lower in BPH eggs laid on ir-ibak1 plants than wild-type (WT) plants; moreover, gravid BPH females preferred to oviposit on WT plants over ir-ibak1 plants. The exogenous application of ethephon on ir-ibak1 plants eliminated the BPH oviposition preference between WT and ir-ibak1 plants but had no effect on the hatching rate of BPH eggs. These findings suggest that OsI-BAK1 acts as a negative modulator of defense responses in rice to BPH and that BPH might exploit this modulator for its own benefit.

Suppression of a leucine-rich repeat receptor-like kinase enhances host plant resistance to a specialist herbivore.

The mechanisms by which herbivores induce plant defenses are well studied. However, how specialized herbivores suppress plant resistance is still poorly understood. Here, we discovered a rice (Oryza sativa) leucine-rich repeat receptor-like kinase, OsLRR-RLK2, which is induced upon attack by gravid females of a specialist piercing-sucking herbivore, the brown planthopper (BPH, Nilaparvata lugens). Silencing OsLRR-RLK2 decreases the constitutive activity of mitogen-activated protein kinase (OsMPK6) and alters BPH-induced transcript levels of several defense-related WRKY transcription factors. Moreover, silencing OsLRR-RLK2 reduces BPH-induction of jasmonic acid and ethylene but promotes the biosynthesis of both elicited salicylic acid and H2 O2 ; silencing also enhances the production of volatiles emitted from rice plants infested with gravid BPH females. These changes decrease BPH preference and performance in the glasshouse and the field. These findings suggest that OsLRR-RLK2, by regulating the plant's defense-related signaling profile, increases the susceptibility of rice to BPH, and that BPH infestation influences the expression of OsLRR-RLK2, suppressing the resistance of rice to BPH.

OsLRR-RLK1, an early responsive leucine-rich repeat receptor-like kinase, initiates rice defense responses against a chewing herbivore.

Plants are constantly exposed to a variety of environmental stresses, including herbivory. How plants perceive herbivores on a molecular level is poorly understood. Leucine-rich repeat receptor-like kinases (LRR-RLKs), the largest subfamily of RLKs, are essential for plants to detect external stress signals, and may therefore also be involved in herbivore perception. Here, we employed RNA interference silencing, phytohormone profiling and complementation, as well as herbivore resistance assays, to investigate the requirement of an LRR-RLK for the initiation of rice (Oryza sativa) defenses against the chewing herbivore striped stem borer (SSB) Chilo suppressalis. We discovered a plasma membrane-localized LRR-RLK, OsLRR-RLK1, whose transcription is strongly up-regulated by SSB attack and treatment with oral secretions of Spodoptera frugiperda. OsLRR-RLK1 acts upstream of mitogen-activated protein kinase (MPK) cascades, and positively regulates defense-related MPKs and WRKY transcription factors. Moreover, OsLRR-RLK1 is a positive regulator of SSB-elicited, but not wound-elicited, levels of jasmonic acid and ethylene, trypsin protease inhibitor activity and plant resistance towards SSB. OsLRR-RLK1 therefore plays an important role in herbivory-induced defenses of rice. Given the well-documented role of LRR-RLKs in the perception of stress-related molecules, we speculate that OsLRR-RLK1 may be involved in the perception of herbivory-associated molecular patterns.

Identification and knockout of a herbivore susceptibility gene enhances planthopper resistance and increases rice yield.

Brown planthoppers (Nilaparvata lugens) and white-backed planthoppers (Sogatella furcifera) are among the most destructive pests on rice. However, plant susceptibility genes have not yet been exploited for crop protection. Here we identified a leucine-rich repeat protein, OsLRR2, from susceptible rice varieties that facilitates infestation by brown planthopper N. lugens. Field trials showed that knockout of OsLRR2 significantly reduced BPH infestation and enhanced natural biological control by attracting natural enemies. Yield of a susceptible variety was increased by 18% in insecticide-treated plots that eliminated planthoppers and by 25% in untreated plots. These findings underscore the pivotal role of OsLRR2, offering a promising pathway for pest population suppression and rice yield increase.

Overexpression of a Cytosolic 6-Phosphogluconate Dehydrogenase Gene Enhances the Resistance of Rice to Nilaparvata lugens.

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.