Huan Liu

The berries of Seabuckthorn (<italic>Hippophae rhamnoides</italic>L.) are traditional medicinal foods that have been used by Tibetans and Mongolians for thousands of years.

Aphids secrete diverse repertoires of salivary effectors into host plant cells to promote infestation by modulating plant defence. The greenbug Schizaphis graminum is an important cereal aphid worldwide. However, the secreted effectors of S. graminum are still uncharacterized. Here, 76 salivary proteins were identified from the watery saliva of S. graminum using transcriptome and proteome analyses. Among them, a putative salivary effector Sg2204 was significantly up-regulated during aphid feeding stages, and transient overexpression of Sg2204 in Nicotiana benthamiana inhibited cell death induced by BAX or INF1. Delivering Sg2204 into wheat via the type III secretion system of Pseudomonas fluorescens EtAnH suppressed pattern-triggered immunity (PTI)-associated callose deposition. The transcript levels of jasmonic acid (JA)- and salicylic acid (SA)-associated defence genes of wheat were significantly down-regulated, and the contents of both JA and SA were also significantly decreased after delivery of Sg2204 into wheat leaves. Additionally, feeding on wheat expressing Sg2204 significantly increased the weight and fecundity of S. graminum and promoted aphid phloem feeding. Sg2204 was efficiently silenced via spray-based application of the nanocarrier-mediated transdermal dsRNA delivery system. Moreover, Sg2204-silenced aphids induced a stronger wheat defence response and resulted in negative impacts on aphid feeding behaviour, survival and fecundity. Silencing of Sg2204 homologues from four aphid species using nanocarrier-delivered dsRNA also significantly reduced aphid performance on host plants. Thus, our study characterized the salivary effector Sg2204 of S. graminum involved in promoting host susceptibility by suppressing wheat defence, which can also be regarded as a promising RNAi target for aphid control.

Aphid salivary proteins are critical in modulating plant defence responses. Grain aphid Sitobion miscanthi is an important wheat pest worldwide. However, the molecular basis for the regulation of the plant resistance to cereal aphids remains largely unknown. Here, we show that SmCSP4, a chemosensory protein from S. miscanthi saliva, is secreted into wheat plants during aphid feeding. Delivery of SmCSP4 into wheat leaves activates salicylic acid (SA)-mediated plant defence responses and subsequently reduces aphid performance by deterring aphid feeding behaviour. In contrast, silencing SmCSP4 gene via nanocarrier-mediated RNAi significantly decreases the ability of aphids to activate SA defence pathway. Protein-protein interaction assays showed that SmCSP4 directly interacts with wheat transcriptional factor TaWRKY76 in plant nucleus. Furthermore, TaWRKY76 directly binds to the promoter of SA degradation gene Downy Mildew Resistant 6 (DMR6) and regulates its gene expression as transcriptional activator. SmCSP4 secreted by aphids reduces the transcriptional activation activity of TaWRKY76 on DMR6 gene expression, which is proposed to result in increases of SA accumulation and enhanced plant immunity. This study demonstrated that SmCSP4 acts as salivary elicitor that is involved in activating SA signalling defence pathway of wheat by interacting with TaWRKY76, which provide novel insights into aphid-cereal crops interactions and the molecular mechanism on induced plant immunity.

Aphid salivary effectors play important roles in modulating plant defense responses. The grain aphid Sitobion miscanthi is one of the most economically important cereal aphids worldwide. However, little information is available on the identification and functional analysis of salivary effectors of S. miscanthi. In this study, a candidate salivary effector Sm9723 was identified, which was specifically expressed in aphid salivary glands and highly induced during the aphid feeding phase. Transient overexpression of Sm9723 in Nicotiana benthamiana suppressed BAX and INF1-induced cell death. Further, Sm9723 overexpression inhibited N. benthamiana defense responses by reducing pattern-triggered immunity associated callose deposition and expression levels of jasmonic and salicylic acid-associated defense genes. In addition, the salivary effector Sm9723 of S. miscanthi was effectively silenced through nanocarrier-mediated dsRNA delivery system. After silencing Sm9723, fecundity and survival of S. miscanthi decreased significantly, and the aphid feeding behavior was also negatively affected. These results suggest salivary effector Sm9723 is involved in suppressing plant immunity and is essential in enabling aphid virulence, which could be applied as potential target gene for RNAi-mediated pest control of S. miscanthi.