BackgroundTreatment of plants with HrpNEa, a protein of harpin group produced by Gram-negative plant pathogenic bacteria, induces plant resistance to insect herbivores, including the green peach aphid Myzus persicae, a generalist phloem-feeding insect. Under attacks by phloem-feeding insects, plants defend themselves using the phloem-based defense mechanism, which is supposed to involve the phloem protein 2 (PP2), one of the most abundant proteins in the phloem sap. The purpose of this study was to obtain genetic evidence for the function of the Arabidopsis thaliana (Arabidopsis) PP2-encoding gene AtPP2-A1 in resistance to M. persicae when the plant was treated with HrpNEa and after the plant was transformed with AtPP2-A1.ResultsThe electrical penetration graph technique was used to visualize the phloem-feeding activities of apterous agamic M. persicae females on leaves of Arabidopsis plants treated with HrpNEa and an inactive protein control, respectively. A repression of phloem feeding was induced by HrpNEa in wild-type (WT) Arabidopsis but not in atpp2-a1/E/142, the plant mutant that had a defect in the AtPP2-A1 gene, the most HrpNEa-responsive of 30 AtPP2 genes. In WT rather than atpp2-a1/E/142, the deterrent effect of HrpNEa treatment on the phloem-feeding activity accompanied an enhancement of AtPP2-A1 expression. In PP2OETAt (AtPP2-A1-overexpression transgenic Arabidopsis thaliana) plants, abundant amounts of the AtPP2-A1 gene transcript were detected in different organs, including leaves, stems, calyces, and petals. All these organs had a deterrent effect on the phloem-feeding activity compared with the same organs of the transgenic control plant. When a large-scale aphid population was monitored for 24 hours, there was a significant decrease in the number of aphids that colonized leaves of HrpNEa-treated WT and PP2OETAt plants, respectively, compared with control plants.ConclusionsThe repression in phloem-feeding activities of M. persicae as a result of AtPP2-A1 overexpression, and as a deterrent effect of HrpNEa treatment in WT Arabidopsis rather than the atpp2-a1/E/142 mutant suggest that AtPP2-A1 plays a role in plant resistance to the insect, particularly at the phloem-feeding stage. The accompanied change of aphid population in leaf colonies suggests that the function of AtPP2-A1 is related to colonization of the plant.

AtMYB44 is a transcription factor that functions in association with the ethylene-signaling pathway in Arabidopsis thaliana. The pathway depends on ETHYLENE INSENSITIVE2 (EIN2), an essential component of ethylene signaling, to regulate defense responses in the plant following treatment with HrpN(Ea), a harpin protein from a bacterial plant pathogen. Here, we show that AtMYB44 regulates induced expression of the EIN2 gene in HrpN(Ea)-treated Arabidopsis plants. A HrpN(Ea) and ethylene-responsive fragment of the AtMYB44 promoter is sufficient to support coordinate expression of AtMYB44 and EIN2 in specific transgenic Arabidopsis. In the plant, the AtMYB44 protein localizes to nuclei and binds the EIN2 promoter; the HrpN(Ea) treatment promotes AtMYB44 production, binding activity, and transcription of AtMYB44 and EIN2. AtMYB44 overexpression results in increased production of the AtMYB44 protein and the occurrence of AtMYB44-EIN2 interaction under all genetic backgrounds of wild-type Arabidopsis and the etr1-1, ein2-1, ein3-1, and ein5-1 mutants, which have defects in the ethylene receptor ETR1 and the signal regulators EIN2, EIN3, and EIN5. However, AtMYB44 overexpression leads to enhanced EIN2 expression only under backgrounds of wild type, ein3-1, and ein5-1 but not etr1-1 and ein2-1, suggesting that ethylene perception is necessary to the regulation of EIN2 transcription by AtMYB44.

The harpin protein HrpN Ea induces Arabidopsis resistance to the green peach aphid by activating the ethylene signalling pathway and by recruiting EIN2, an essential regulator of ethylene signalling, for a defence response in the plant. We investigated 37 ethylene-inducible Arabidopsis transcription factor genes for their effects on the activation of ethylene signalling and insect defence. Twenty-eight of the 37 genes responded to both ethylene and HrpN Ea, and showed either increased or inhibited transcription, while 18 genes showed increased transcription not only by ethylene but also by HrpN Ea. In response to HrpN Ea, transcription levels of 22 genes increased, with AtMYB44 being the most inducible, six genes had decreased transcript levels, and nine remained unchanged. When Arabidopsis mutants previously generated by mutagenicity at the 37 genes were surveyed, 24 mutants were similar to the wild type plant while four mutants were more resistant and nine mutants were more susceptible than wild type to aphid infestation. Aphid-susceptible mutants showed a greater susceptibility for atmyb15, atmyb38 and atmyb44, which were generated previously by T-DNA insertion into the exon region of AtMYB15 and the promoter regions of AtMYB38 and AtMYB44. The atmyb44 mutant was the most susceptible to aphid infestation and most compromised in induced resistance. Resistance accompanied the expression of PDF1.2, an ethylene signalling marker gene that requires EIN2 for transcription in wild type but not in atmyb15, atmyb38, and atmyb44, suggesting a disruption of ethylene signalling in the mutants. However, only atmyb44 incurred an abrogation in induced EIN2 expression, suggesting a close relationship between AtMYB44 and EIN2.

SummaryTRANSPARENT TESTA GLABRA (TTG) proteins that contain the WD40 protein interaction domain are implicated in many signalling pathways in plants. The salicylic acid (SA) signalling pathway regulates the resistance of plants to pathogens through defence responses involving pathogenesis-related (PR) gene transcription, activated by the NPR1 (nonexpresser of PR genes 1) protein, which contains WD40-binding domains. We report that tobacco (Nicotiana tabacum) NtTTG2 suppresses the resistance to viral and bacterial pathogens by repressing the nuclear localisation of NPR1 and SA/NPR1-regulated defence in plants. Prevention of NtTTG2 protein production by silencing of the NtTTG2 gene resulted in the enhancement of resistance and PR gene expression, but NtTTG2 overexpression or NtTTG2 protein overproduction caused the opposite effects. Concurrent NtTTG2 and NPR1 gene silencing or NtTTG2 silencing in the absence of SA accumulation compensated for the compromised defence as a result of the NPR1 single-gene silencing or the absence of SA. However, NtTTG2 did not interact with NPR1 but was able to modulate the subcellular localisation of the NPR1 protein. In the absence of NtTTG2 production NPR1 was found predominantly in the nucleus and the PR genes were expressed. By contrast, when NtTTG2 accumulated in transgenic plants, a large proportion of NPR1 was retained in the cytoplasm and the PR genes were not expressed. These results suggest that NtTTG2 represses SA/NPR1-regulated defence by sequestering NPR1 from the nucleus and the transcriptional activation of the defence-response genes.

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Harpin proteins secreted by phytopathogenic bacteria have been shown to activate the plant defense pathway, which involves transduction of a hydrogen peroxide (H(2)O(2)) signal generated in the apoplast. However, the way in which harpins are recognized in the pathway and what role the apoplastic H(2)O(2) plays in plant defenses are unclear. Here, we examine whether the cellular localization of Hpa1(Xoo), a harpin protein produced by the rice bacterial leaf blight pathogen, impacts H(2)O(2) production and pathogen resistance in Arabidopsis thaliana. Transformation with the hpa1 (Xoo) gene and hpa1 (Xoo) fused to an apoplastic localization signal (shpa1 (Xoo)) generated h pa1 (Xoo)- and sh pa1 (Xoo)-expressing transgenic A . t haliana (HETAt and SHETAt) plants, respectively. Hpa1(Xoo) was associated with the apoplast in SHETAt plants but localized inside the cell in HETAt plants. In addition, Hpa1(Xoo) localization accompanied H(2)O(2) accumulation in both the apoplast and cytoplasm of SHETAt plants but only in the cytoplasm of HETAt plants. Apoplastic H(2)O(2) production via nicotinamide adenine dinucleotide phosphate (NADPH) oxidase (NOX) located in the plasma membrane is a common feature of plant defenses. In SHETAt plants, H(2)O(2) was generated in apoplasts in a NOX-dependent manner but accumulated to a greater extent in the cytoplasm than in the apoplast. After being applied to the wild-type plant, Hpa1(Xoo) localized to apoplasts and stimulated H(2)O(2) production as in SHETAt plants. In both plants, inhibiting apoplastic H(2)O(2) generation abrogated both cytoplasmic H(2)O(2) accumulation and plant resistance to bacterial pathogens. These results suggest the possibility that the apoplastic H(2)O(2) is subject to a cytoplasmic translocation for participation in the pathogen defense.

The cDNA from Nicotiana tabacum encoding Putrescine N-methyltransferase (PMT), which catalyzes the first committed step in the biosynthesis of tropane alkaloids, has been introduced into the genome of a scopolamine-producing Hyoscyamus niger mediated by the disarmed Agrobacterium tumefaciens strain C58C1, which also carries Agrobacterium rhizogenes Ri plasmid pRiA4, and expressed under the control of the CaMV 35S promoter. Hairy root lines transformed with pmt presented fivefold higher PMT activity than the control, and the methylputrescine (MPUT) levels of the resulting engineered hairy roots increased four to fivefold compared to the control and wild-type roots, but there was no significant increase in tropane alkaloids. However, after methyl jasmonate (MeJA) treatment, a considerable increase of PMTase and endogenous H6Hase as well as an increase in scopolamine content was found either in the transgenic hairy roots or the control. The results indicate that hairy root lines over-expressing pmt have a high capacity to synthesize MPUT, whereas their ability to convert hyoscyamine into scopolamine is very limited. Exposure to MeJA strongly stimulated both polyamine and tropane biosynthesis pathways and elicitation led to more or less enhanced production simultaneously.

and likely recurrence. Therefore, it is necessary to develop more effective treatment methods. With the in depth study of the various dynamics and heterogeneous characteristics of cancer, a new type of "nanobiomedicine" that combines imaging diagnostic methods and effective tumorremoving therapeutic drugs have garnered significant attention. Nanobiomedicine refers to the application of nanomaterials in the field of biomedicine for the diagnosis and treatment of diseases. In recent years, many nanomaterials combined with biological therapy have appeared. Among them, 2D nanomaterials have stood out in the diagnosis and treatment of cancer and have become one of the most valuable materials for nanomedicine applications. [1][2][3] 2D nanomaterials have attracted considerable attention because of their fewlayer nanostructures, large lateral planar structures, and unique physical and chemical properties. In recent years, many studies have been conducted on various 2D nanomaterial systems for biomedical applications. 2D nanomaterials include graphene, transition metal disulfide, black phosphorus, layered double hydroxide, palladium, hexagonal boron nitride, 2D transition metal-carbon (nitrogen) compounds, and other materials. [2] Their surface area, unique surface chemical functions, and inherent optical properties make them suitable for a variety of applications in nanobiomedicine. Previous studies showed that 2D nanomaterials can be used for drug and gene loading, drug delivery, [4,5] photothermal therapy (PTT), [6] diagnostic imaging, [7] and other applications. [8] MXenes have been extensively studied as novel 2D nanomaterials. They are a large class of 2D nanomaterials with planar structures, including transition metal carbides, nitrides, or carbonitrides. In 2011, Ti 3 C 2 T x , which was the earliest synthesized MXene material, developed from the MAX phase of Ti 3 AlC 2 . [3] MXenes show great potential for application in the field of biomedicine, including in PTT, diagnostic imaging, and biosensing, because the 2D nano-planar structure renders them rich in anchor points. Thus, they act as excellent drug delivery agents for carrying cancer-targeting drug molecules or other proteins. Besides, MXenes exhibit strong near-infrared (NIR) absorption, electronic transparency, and X-ray attenuation abilities. They have adjustable designs and strategies for the synthesis of the original structure of the MAX phase. MXenes can also reduce cytotoxicity and enhance biological stability and histocompatibility through surface modification, which provides the possibility of in vivo applications. [9] Research on 2D nanomaterials is still in its early stages. Most studies have focused on elucidating the unique properties of the materials, whereas only few reports have described the biomedical applications of 2D nanomaterials. Recently, important questions about the interaction of 2D MXene nanomaterials with biological components have been raised. 2D MXenes are monolayer atomic nanosheets derived from MAX phase ceramics. As a new t...