Protein Elicitor EsxA Induces Resistance to Seedling Blight and PR Genes Differential Transcription in Rice

Protein elicitors can induce plant systemic resistance to pathogens. In an earlier study, we cloned an EsxA gene from the plant growth-promoting rhizobacterium Paenibacillus terrae NK3-4 and expressed it in Pichia pastoris. In addition to being important for the pathogenicity of animal pathogens, EsxA can also induce an immune response in animals. While, we found the exogenously expressed EsxA has the activity of elicitor, which can trigger hypersensitive response and reactive oxygen species burst in leaves as well as enhanced rice plant growth. The effects of EsxA on seedling blight (Fusarium oxysporum) resistance and gene transcription, including pathogenesis-related (PR) genes in rice were evaluated. The germination rate was 95.0% for seeds treated with EsxA and then inoculated with F. oxysporum, which was 2.8-times higher than that of F. oxysporum-infected control seeds that were not treated with EsxA (Con). The buds and roots of EsxA-treated seedlings were 2.4- and 15.9-times longer than those of Con seedlings. The plants and roots of seedlings dipped in an EsxA solution and then inoculated with F. oxysporum were longer than those of the Con seedlings. Theplant length, number of total roots, and number of white roots were respectively 23.2%, 1.74-times, and 7.42-times greater for the seedlings sprayed with EsxA and then inoculated with F. oxysporum than for the Con seedlings. The EsxA induction efficiency (spray treatment) on seedling blight resistance was 60.9%. The transcriptome analysis revealed 1137 and 239 rice genes with EsxA-induced up-regulated and down-regulated transcription levels, respectively. At 48 h after the EsxA treatment, the transcription of 611 and 160 genes was up-regulated and down-regulated, respectively, compared with the transcription levels for the untreated control at the same time-point. Many disease resistance-related PR genes had up-regulated transcription levels. The qPCR data were consistent with the transcriptome sequencing results. EsxA triggered rice ISR to seedling blight and gene differential transcription, including the up-regulated transcription of rice PR genes. These findings may be relevant for the use of EsxA as a protein elicitor to control plant diseases.

Biocontrol Potential of Purified Elicitor Protein PeBL1 Extracted from Brevibacillus laterosporus Strain A60 and Its Capacity in the Induction of Defense Process against Cucumber Aphid (Myzus persicae) in Cucumber (Cucumis sativus)

The Cucumber aphid (Myzus persicae), a destructive cucumber aphid usually managed by chemical pesticides, is responsible for enormous annual agricultural losses. A protein elicitor, PeBL1, was investigated in the present work for its ability to induce a defense response against M. persicae in cucumber. The rates of population growth (Intrinsic rate of increase) of M. persicae (second and third generations) decreased with PeBL1-treated cucumber seedlings as compared to positive (water) and negative 70.58 μg mL−1 controls (50 mM Tris-HCl, pH 8.0). In an assay on host selection, M. persicae had a preference for colonizing control plants as compared to the PeBL1-treated cucumber seedlings. The nymphal development time of the aphid was extended with the PeBL1-treated cucumber seedlings. Likewise, fecundity was reduced, with less offspring produced in the PeBL1-treated cucumber seedlings as compared to the positive (water) and negative 70.58 μg mL−1 controls (50 mM Tris-HCl, pH 8.0). The cucumber leaves treated with PeBL1 had a hazardous surface environment for M. persicae, caused by trichomes and wax formation. Jasmonic acid (JA), salicylic acid (SA), and ethylene (ET) levels were significantly higher, exhibiting significant accumulation in the PeBL1-treated cucumber seedlings. The following results showed that PeBL1 considerably altered the height of the cucumber plant and the surface structure of the leaves to minimize M. persicae reproduction, and it prevented colonization. Defensive processes also included the activation of pathways (JA, SA, and ET). This study provides evidence of biocontrol for the use of PeBL1 in cucumber defense against M. persicae.

Putative Role of a Yet Uncharacterized Protein Elicitor PeBb1 Derived from Beauveria bassiana ARSEF 2860 Strain against Myzus persicae (Homoptera: Aphididae) in Brassica rapa ssp. pekinensis

This study reports the characterization of protein elicitor PeBb1 derived from entomopathogenic fungus Beauveria bassiana ARSEF-2860 strain and its putative role in induced systemic resistance in Brassica rapa ssp. pekinensis against green peach aphid Myzus persicae. The sequence of purified elicitor protein was matched with the genomic sequence of a hypothetical protein BBA_10269 from B. bassiana ARSEF-2860 (GenBank Accession No. XP_008603588.1). The protein-encoding gene PeBb1 contained 534 bp cDNA encoding a polypeptide of 177 amino acids with a molecular mass of 19 kDa. The recombinant elicitor protein was expressed in Escherichia coli using pET-28a (+) expression vector and induced necrosis in the leaves of tobacco. The effects of elicitor protein on aphid M. persicae was determined by applying three different concentrations of PeBb1 (i.e., 26, 35, 53 μM) on B. rapa plants at 4-leaf stage and the treated plants were exposed to newly emerged (0–6 h old) apterous adult aphids. Bioassay results showed significant (p < 0.05) sub-lethal effects of the exogenous application of PeBb1 elicitor on M. persicae. Moreover, the RT-qPCR gene expression analyses showed a significant up-regulation of most of the key genes linked to ethylene (ET)- and jasmonic acid (JA)-associated plant defense pathways in elicitor-treated plants. These results not only recommend the putative utilization of PeBb1 elicitor protein in future biological pest control strategies against phloem-feeding insect pests such as M. persicae, but also help in better comprehension of the mechanisms through which beneficial fungi trigger the induced plant resistance.

Protein Elicitor PeBL1 of Brevibacillus laterosporus Enhances Resistance Against Myzus persicae in Tomato

Myzus persicae, a destructive aphid of tomato usually managed by chemical pesticides, is responsible for huge annual losses in agriculture. In the current work, a protein elicitor, PeBL1, was investigated for its capacity to induce a defense response against M. persicae in tomato. Population growth rates of M. persicae (second and third generation) decreased with PeBL1 treatments as compared with controls. In a host selection assay, M. persicae showed preference for colonizing control plants as compared to tomato seedlings treated with PeBL1. Tomato leaves treated with PeBL1 gave rise to a hazardous surface environment for M. persicae due to formation of trichomes and wax. Jasmonic acid (JA), salicylic acid (SA), and ethylene (ET) showed significant accumulation in tomato seedlings treated by PeBL1. The following results showed that PeBL1 significantly modified the tomato leaf surface structure to reduce reproduction and deter colonization by M. persicae. Defense processes also included activation of JA, SA, and ET pathways. The study provides evidence for use of PeBL1 in the protection of tomato from M. persicae.

Response mechanism of hypocrellin colorants biosynthesis by Shiraia bambusicola to elicitor PB90

The valuable medicine Shiraia bambusicola P. Henn. and its major active substance hypocrellin exert unique curative effects on skin diseases, diabetes, and cancers. The wild S. bambusicola is endangered due to its harsh breeding conditions and long growth cycle. It is one of the effective ways to utilize the resources sustainably to produce hypocrellin by fermentation of S. bambusicola. PB90 is a protein elicitor isolated from Phytophthora boehmeriae to induce the useful metabolites production in fungi. In this work, PB90 was selected to promote the synthesis hypocrellin by S. bambusicola. To evaluate the effect of PB90 on S. bambusicola, it was found that the induced cells showed decreased biomass, increased cell wall permeability, rapid induction of secondary metabolites, and significant increase of some enzyme activities, which confirmed a strong activation of phenylalanine/flavonoid pathways. Studies on signal molecules and gene expression level in S. bambusicola treated with PB90 have found that hydrogen peroxide (H2O2) and nitric oxide (NO) are necessary signal molecules involved in the synthesis of hypocrellin in elicited cells, and increased their signal levels through mutual reaction. We have showed for the first time, the response mechanism of hypocrellin biosynthesis from S. bambusicola to PB90, which may be not only establish a theoretical foundation for the application of PB90 to the mass production of S. bambusicola, but can also motivate further research on the application of PB90 to the conservation and sustainable utilization of other medical fungi.

Protein Elicitor PeaT1 Efficiently Controlled Barley Yellow Dwarf Virus in Wheat

Barley yellow dwarf virus (BYDV), transmitted by the wheat aphid, generates serious wheat yellow dwarf disease and causes great losses in agriculture. Induced resistance has attracted great attention over recent years as a biological method to control plant pathogens and herbivores. Protein elicitor PeaT1 induces defense response in plants against fungi, viruses, and aphids. In this study, wheat seeds and seedlings were soaked and sprayed with 30 μg/mL PeaT1, respectively. Then seedlings were inoculated with BYDV by viruliferous Schizaphis graminum to detect the control efficiency of PeaT1-induced resistance against BYDV. The control efficiency was over 30% on the 14th and 21st days after the inoculation access period. Quantitative real time polymerase chain reaction (Q-RT-PCR) tests showed that there was less mRNA from the BYDV coat protein in PeaT1-treated wheat seedlings than in the control group. Electrical penetration graph (EPG) tests showed that virus transmission vector S.graminum took a longer time to find probe and feeding sites on PeaT1-treated wheat seedlings. Additionally, PeaT1-treated wheat seedlings gained higher plant height and more chlorophyll a&b. These results showed that PeaT1 efficiently controlled BYDV by inhibiting BYDV proliferation, reducing the virus transmission ability of S. graminum and alleviating the symptoms of dwarfism and yellow colouring caused by BYDV. This study provided a new integrated way to control BYDV biologically.