Specialist aphids feeding causes local activation of salicylic and jasmonic acid signaling in Arabidopsis veins

Aphids, the phloem sap feeders, probe into leaf tissues and activate a complex network of plant defence responses. Phytohormonal signaling plays a major role in this network; however, the dynamics of the signals spreading is yet to be clarified. Despite the growing knowledge about transcriptomic changes upon infestation, results often differ due to sampling, varying strongly between the tissues collected at the single feeding site, individual leaves, pooled infested leaves, or whole plant rosettes. This study focuses on activation of salicylic and jasmonic acid signals in Arabidopsis leaves during infestation by cabbage aphid (Brevicoryne brassicae) in high spatio-temporal resolution. We used genetically encoded fluorescent biosensors, histochemistry and qRT-PCR to precisely map activation of distinct branches of phytohormonal signaling. We found a rapid induction of salicylic and jasmonic acid signaling markers in cells surrounding stylet puncture, co-localizing with callose deposition. For both PR1 and JAZ10 we detected activation at 24 hpi, increasing and spreading along the veins until 72 hpi and, to a lesser extent, within the epidermal pavement cells. The SA signaling wave appeared in parallel with JA-associated, and continued to increase in time. Our results first show a local activation of SA- and JA-related responses after stylet penetration of Arabidopsis leaves and bring a detailed insight into the spatio-temporal complexity of plant defence activation during specialist aphid attack.

Rickettsia increases its infection and spread in whitefly populations by manipulating the defense patterns of the host plant

ABSTRACT The whitefly Bemisia tabaci is a destructive agricultural pest that frequently harbors various species of secondary symbionts including Rickettsia. Previous studies have revealed that the infection of Rickettsia can improve whitefly performance on food plants; however, to date, no evidence has shown, if, and how, Rickettsia manipulates the plant-insect interactions. In the current study, the effects of Rickettsia persistence on the induced plant defenses and the consequent performance of whitefly B. tabaci were investigated. Results revealed that Rickettsia can be transmitted into plants via whitefly feeding and remain alive within the cotton plants for at least 2 weeks. The different expression genes of cotton plants were mostly concentrated in the phytohormone signaling pathways, the marker genes of jasmonic-acid signaling pathway (AOC, AOS, LOX, MYC2) were significantly downregulated, while the marker genes of the salicylic-acid signaling pathway (WRKY70, PR-1) were upregulated. Biological experiments revealed that the fecundity of Rickettsia negative B. tabaci significantly increased when they fed on Rickettsia-persistent cotton plants. Taken together, we provide experimental evidence that the persistence of Rickettsia and its induced defense responses in cotton plants can increase the fitness of whitefly and, by this, Rickettsia may increase its infection and spread within its whitefly host.

Integrated transcriptomic and metabolomic analyses reveal regulation of terpene biosynthesis in the stems of Sindora glabra

Sesquiterpenes are important defensive secondary metabolites that are synthesized in various plant organs. Methyl jasmonate (MeJA) plays a key role in plant defense responses and secondary metabolism. Sindora glabra produces abundant sesquiterpenes in their trunks and was subjected to investigation after MeJA treatment in order to characterize the molecular mechanisms underlying the regulation of sesquiterpene biosynthesis in plant stems and further our understanding of oleoresin production in trees. A total of 14 types of sesquiterpenes in the stems of mature S. glabra trees were identified. The levels of two sesquiterpenes, α-copaene and β-caryophyllene, significantly increased after MeJA treatment. Differentially expressed genes involved in terpenoid backbone biosynthesis were significantly enriched over time, while the expression of JAZ genes involved in the jasmonic acid signaling pathway and TGA genes involved in the salicylic acid signaling pathway was significantly enriched at later time points after treatment. Two new terpene synthase genes, SgSTPS4 and SgSTPS5, were also identified. Following MeJA treatment, the expression levels of SgSTPS1, SgSTPS2, and SgSTPS4 decreased, while SgSTPS5 expression increased. The major enzymatic products of SgSTPS4 were identified as β-elemene and cyperene, while SgSTPS5 was identified as a bifunctional mono/sesquiterpene synthase that could catalyze FPP to produce nine types of sesquiterpenes, including α-copaene and β-caryophyllene, while SgSTPS5 could also use GPP to produce geraniol. Dramatic changes in the amounts of α-copaene and β-caryophyllene in response to MeJA were correlated with transcriptional expression changes of SgSTPS5 in the wood tissues. In addition, the transcription factors MYB, NAC, ARF, WRKY, MYC, ERF, and GRAS were co-expressed with terpene biosynthesis genes and might potentially regulate terpene biosynthesis. Metabolite changes were further investigated with UPLC-TOF/MS following MeJA treatment. These results contribute to the elucidation of the molecular mechanisms of terpene biosynthesis and regulation as well as to the identification of candidate genes involved in these processes.

Heterologous microProtein expression identifies LITTLE NINJA, a dominant regulator of jasmonic acid signaling

MicroProteins are small, often single-domain proteins that are sequence-related to larger, often multidomain proteins. Here, we used a combination of comparative genomics and heterologous synthetic misexpression to isolate functional cereal microProtein regulators. Our approach identified LITTLE NINJA (LNJ), a microProtein that acts as a modulator of jasmonic acid (JA) signaling. Ectopic expression ofLNJinArabidopsisresulted in stunted plants that resembled the decupleJAZ(jazD) mutant. In fact, comparing the transcriptomes of transgenicLNJoverexpressor plants andjazDrevealed a large overlap of deregulated genes, suggesting that ectopicLNJexpression altered JA signaling. Transgenic Brachypodium plants with elevatedLNJexpression levels showed deregulation of JA signaling as well and displayed reduced growth and enhanced production of side shoots (tiller). This tillering effect was transferable between grass species, and overexpression ofLNJin barley and rice caused similar traits. We used a clustered regularly interspaced short palindromic repeats (CRISPR) approach and created a LNJ-like protein inArabidopsisby deleting parts of the coding sentence of theAFP2gene that encodes a NINJA-domain protein. Theseafp2-crisprmutants were also stunted in size and resembledjazD. Thus, similar genome-engineering approaches can be exploited as a future tool to create LNJ proteins and produce cereals with altered architectures.