The use of PTI-marker genes to identify novel compounds that establish induced resistance in rice

Compounds that establish induced resistance (IR) in plants are promising alternatives for the pesticides that are progressively being banned worldwide. Screening platforms to identify IR-establishing compounds have been developed, but none were specifically designed for monocot plants. Here, we propose the use of an RT-qPCR screening platform, based on conserved immunity marker genes of rice as proxy for IR induction. Central regulators of biotic stress responses of rice were identified with a weighted gene co-expression network analysis (WGCNA), using more than 350 microarray datasets of rice under various sorts of biotic stress. Candidate genes were narrowed down to six immunity marker genes, based on consistent association with pattern-triggered immunity (PTI), both in rice plants as in rice cell suspension cultures (RCSCs). By monitoring the expression of these genes in RCSCs upon treatment with candidate IR-inducing compounds, we showed that our marker genes can predict IR induction in rice. Diproline, a novel IR-establishing compound for monocots that was detected with these marker genes, was shown to induce rice resistance against root-knot nematodes, without fitness costs. Gene expression profiling of the here-described PTI-marker genes can be executed on fully-grown plants or in RCSCs, providing a novel and versatile tool to predict IR induction

Altering the sex pheromone cyclo(L-Pro-L-Pro) of the diatom Seminavis robusta towards a chemical probe

As a major group of algae, diatoms are responsible for a substantial part of the primary production on the planet. Pennate diatoms have a predominantly benthic lifestyle and are the most species-rich diatom group, with members of the raphid clades being motile and generally having heterothallic sexual reproduction. It was recently shown that the model species Seminavis robusta uses multiple sexual cues during mating, including cyclo(l-Pro-l-Pro) as an attraction pheromone. Elaboration of the pheromone-detection system is a key aspect in elucidating pennate diatom life-cycle regulation that could yield novel fundamental insights into diatom speciation. This study reports the synthesis and bio-evaluation of seven novel pheromone analogs containing small structural alterations to the cyclo(l-Pro-l-Pro) pheromone. Toxicity, attraction, and interference assays were applied to assess their potential activity as a pheromone. Most of our analogs show a moderate-to-good bioactivity and low-to-no phytotoxicity. The pheromone activity of azide- and diazirine-containing analogs was unaffected and induced a similar mating behavior as the natural pheromone. These results demonstrate that the introduction of confined structural modifications can be used to develop a chemical probe based on the diazirine- and/or azide-containing analogs to study the pheromone-detection system of S. robusta

Bioactivity-guided isolation of rosmarinic acid as the principle bioactive compound from the butanol extract of Isodon rugosus against pea aphid, Acyrthosiphon pisum

Aphids are agricultural pest insects that transmit viruses and cause feeding damage on a global scale. Current pest control involving the excessive use of synthetic insecticides over decades has led to multiple forms of aphid resistance to most classes of insecticides. In nature, plants produce secondary metabolites during their interaction with insects and these metabolites can act as toxicants, antifeedants, anti-oviposition agents and deterrents towards the insects. In a previous study, we demonstrated that the butanol fraction from a crude methanolic extract of an important plant species, Isodon rugosus showed strong insecticidal activity against the pea aphid, Acyrthosiphon pisum. It was however not known as which compound was responsible for such activity. To further explore this finding, current study aimed to exploit a bioactivity-guided strategy to isolate and identify the active compound in the butanol fraction of I. rugosus. As such, reversed-phase flash chromatography, acidic extraction and different spectroscopic techniques were used to isolate and identify the new compound, rosmarinic acid as the bioactive compound in I. rugosus. Insecticidal activity of rosmarinic acid was carried out using standard protocols on A. pisum. The data was analyzed using qualitative and quantitative statistical approaches. Considering that a very low concentration of this compound (LC90 = 5.4 ppm) causes significant mortality in A. pisum within 24 h, rosmarinic acid could be exploited as a potent insecticide against this important pest insect. Furthermore, I. rugosus is already used for medicinal purposes and rosmarinic acid is known to reduce genotoxic effects induced by chemicals, hence it is expected to be safer compared to the current conventional pesticides. While this study highlights the potential of I. rugosus as a possible biopesticide source against A. pisum, it also provides the basis for further exploration and development of formulations for effective field application

Bioactivity-guided isolation of rosmarinic acid as the principal bioactive compound from the butanol extract of Isodon rugosus against the pea aphid, Acyrthosiphon pisum

Aphids are agricultural pest insects that transmit viruses and cause feeding damage on a global scale. Current pest control practices involving the excessive use of synthetic insecticides over many years have resulted in aphid resistance to a number of pesticides. In nature, plants produce secondary metabolites during their interaction with insects and these metabolites can act as toxicants, antifeedants, anti-oviposition agents and deterrents towards the insects. In a previous study, we demonstrated that the butanol fraction from a crude methanolic extract of an important plant species, Isodon rugosus showed strong insecticidal activity against the pea aphid, Acyrthosiphon pisum. To further explore this finding, the current study aimed to exploit a bioactivity-guided strategy to isolate and identify the active compound in the butanol fraction of I. rugosus. As such, reversed-phase flash chromatography, acidic extraction and different spectroscopic techniques were used to isolate and identify the new compound, rosmarinic acid, as the bioactive compound in I. rugosus. Insecticidal potential of rosmarinic acid against A. pisum was evaluated using standard protocols and the data obtained was analyzed using qualitative and quantitative statistical approaches. Considering that a very low concentration of this compound (LC90 = 5.4 ppm) causes significant mortality in A. pisum within 24 h, rosmarinic acid could be exploited as a potent insecticide against this important pest insect. Furthermore, I. rugosus is already used for medicinal purposes and rosmarinic acid is known to reduce genotoxic effects induced by chemicals, hence it is expected to be safer compared to the current conventional pesticides. While this study highlights the potential of I. rugosus as a possible biopesticide source against A. pisum, it also provides the basis for further exploration and development of formulations for effective field application

Bioactivity-guided isolation of rosmarinic acid as the principal bioactive compound from the butanol extract of Isodon rugosus against the pea aphid, Acyrthosiphon pisum (vol 14, e0215048, 2019)

[This corrects the article DOI: 10.1371/journal.pone.0215048.]

Mating type specific transcriptomic response to sex inducing pheromone in the pennate diatom Seminavis robusta

Sexual reproduction is a fundamental phase in the life cycle of most diatoms. Despite its role as a source of genetic variation, it is rarely reported in natural circumstances and its molecular foundations remain largely unknown. Here, we integrate independent transcriptomic datasets to prioritize genes responding to sex inducing pheromones (SIPs) in the pennate diatomSeminavis robusta. We observe marked gene expression changes associated with SIP treatment in both mating types, including an inhibition of S phase progression, chloroplast division, mitosis, and cell wall formation. Meanwhile, meiotic genes are upregulated in response to SIP, including a sexually induced diatom specific cyclin. Our data further suggest an important role for reactive oxygen species, energy metabolism, and cGMP signaling during the early stages of sexual reproduction. In addition, we identify several genes with a mating type specific response to SIP, and link their expression pattern with physiological specialization, such as the production of the attraction pheromone diproline in mating type - (MT-) and mate-searching behavior in mating type + (MT+). Combined, our results provide a model for early sexual reproduction in pennate diatoms and significantly expand the suite of target genes to detect sexual reproduction events in natural diatom populations

Alkaloids isolated from Ephedra alata : characterization and protective effects against cisplatin-induced liver and kidney injuries in mice

Cisplatin (CP) is one of the most efficacious chemotherapeutic antitumor drugs. Oxidative stress has been proven to be involved in CP-induced toxicity. The aim of this study was designed to assess the protective effects of Ephedra alata alkaloids extract (AE) on liver and kidney injuries induced by CP. The 1H-RMN analysis of AE extract detected the presence of ephedrine, pseudoephedrine, methylephedrine and methylpseudoephedrine. To evaluate the effect of AE extract on CP-toxicity, the mice were administrated with 150 mg/kg of AE for 7 days, and the liver and kidney injury models were established by single intraperitoneal injection of CP (20mg/kg) on the fourth day. Compared with the model group, the activities of aspartate aminotransferase, alanine aminotransferase and the content of creatinine in serum all decreased in mice treated with AE extract. Meanwhile, the activities of superoxide dismutase, catalase increased, and the content of malondialdehyde and DNA damage decreased. In addition, the histopathologic aspects showed that the pathological changes of liver and kidney were found in the model group reduced after treatment with AE. Therefore, AE could reduce the damage of liver and kidney caused by CP by reducing the level of oxidative stress, and improving the antioxidant, capacity of the body

Diproline-induced resistance to parasitic nematodes in the same and subsequent rice generations : roles of iron, nitric oxide and ethylene

Induced resistance (IR) is a plant phenotype characterized by lower susceptibility to biotic challenges upon elicitation by so-called IR stimuli. Earlier, we identified diproline (cyclo(l-Pro-l-Pro)) as IR stimulus that protects rice (Oryza sativa) against the root-knot nematode Meloidogyne graminicola (Mg). In the current study, detailed transcriptome analyses at different time points, and under uninfected and nematode-infected conditions revealed that this rice IR phenotype is correlated with induction of genes related to iron (Fe), ethylene (ET) and reactive oxygen species (ROS)/reactive nitrogen species (RNS) metabolism. An infection experiment under Fe limiting conditions confirmed that diproline-IR is only effective under optimal Fe supply. Although total root Fe levels were not affected in diproline-treated plants, phytosiderophore secretion was found to be induced by this treatment. Experiments on mutant and transgenic rice lines impaired in ET or ROS/RNS metabolism confirmed that these metabolites are involved in diproline-IR. Finally, we provide evidence for transgenerational inheritance of diproline-IR (diproline-TIR), as two successive generations of diproline-treated ancestors exhibited an IR phenotype while themselves never being exposed to diproline. Transcriptome analyses on the offspring plants revealed extensive overlap between the pathways underpinning diproline-IR and diproline-TIR. Although diproline induces significant systemic changes in global DNA methylation levels early after treatment, such changes in DNA methylation were not detected in the descendants of these plants. To our knowledge, this is the first report of TIR in rice and the first transcriptional assessment of TIR in monocots

Insecticidal activity of plant-derived extracts against different economically important pest insects

With the aim of selecting potential botanical insecticides, seven plant extracts (Daphne mucronata (Family: Thymelaeaceae), Tagetes minuta (Asteraceae), Calotropis procera (Apocynaceae), Boenninghausenia albiflora (Rutaceae), Eucalyptus sideroxylon (Myrtaceae), Cinnamomum camphora (Lauraceae) and Isodon rugosus (Lamiaceae)) were screened for their toxic effects against four important agricultural pest insects, each representing a separate insect order; pea aphids of Acyrthosiphon pisum (Hemiptera), fruit flies of Drosophila melanogaster (Diptera), red flour beetles of Tribolium castaneum (Coleoptera), and armyworms of Spodoptera exigua (Lepidoptera). Aphids were the most susceptible insect with 100% mortality observed after 24 h for all the plant extracts tested. Further bioassays with lower concentrations of the plant extracts against aphids, revealed the extracts from I. rugosus (LC50 36 ppm and LC90 102 ppm) and D. mucronata (LC50 126 ppm and LC90 198 ppm) to be the most toxic to aphids. These most active plant extracts were further fractionated into different solvent fractions on polarity basis and their insecticidal activity evaluated. While all the fractions showed considerable mortality in aphids, the most active was the butanol fraction from I. rugosus with an LC50 of 18 ppm and LC90 of 48 ppm. Considering that high mortality was observed in aphids within 24 h of exposure to a very low concentration of the butanol fraction from I. rugosus, we believe this could be exploited and further developed as a potential plant-based insecticide against sucking insect pests, such as aphids