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Current research does not fully elucidate the key compounds and their mechanisms that define the aroma profile of fresh jujube fruits. Therefore, this study conducted a comprehensive analysis of both free and glycosidically bound aroma compounds in fresh jujube fruits of ten cultivars. Utilizing gas chromatography–mass spectrometry (GC-MS), we identified 76 volatile free aroma compounds and 19 glycosidically bound volatile compounds, with esters, aldehydes, and ketones emerging as the predominant volatile compounds in the jujube fruits. Odor activity value (OAV) analysis revealed that the primary aroma profile of the jujubes is characterized by fruity and fatty odors, with β-damascenone being a key contributor to the fruity aroma, and (E)-2-oct-en-1-al and nonanal significantly influencing the fatty aroma. Moreover, the integration of sensory evaluation and partial least squares regression (PLSR) analysis pinpointed octanal, (E)-2-oct-en-1-al, nonanal, β-damascenone, and pentanal as significant contributors to the jujube’s characteristic aroma, while isoamyl acetate was identified as significantly influencing the fatty acid taste. This study not only underscores the complexity of the jujube aroma composition but also highlights the impact of environmental factors on aroma profiles, offering valuable insights into the sensory characteristics of jujube fruits.

In this paper, a tesla-shaped emitter is proposed based on the structure of the “tesla valve” as the source of inspiration, so that the water flow in the channel would produce a variety of energy dissipation phenomena, such as diversion, hedging, and mixing, to explore the hydraulic performance and energy dissipation mechanism of the tesla-shaped emitter. The channel structure parameters were taken as factors, and 16 groups of orthogonal tests were arranged. Based on CFD technology, the pressure–flow relationship curve slope, flow ratio between the main channel and secondary channel, flow field, and head loss of the emitter were calculated and analyzed for different combinations of structural parameters. Based on a significance level α = 0.05 test, the main channel inlet section length (L3) had a significant impact on the curve slope, and the secondary channel length (L1) and main channel inlet section length (L3) had a significant impact on the flow. The multiple linear regression mathematical models between the channel structure parameters and the curve slope and the flow were constructed. The larger the ratio between the main channel and the secondary channel flow, the better the hydraulic performance of the emitter. The channel unit loss coefficient increased linearly with the increase of the emitter inlet pressure, and its value ranged from 4.5769 to 8.1716, with an excellent energy dissipation effect. The hedge mixing of the water flow was the core of the energy dissipation of the tesla-shaped emitter. By appropriately increasing the inlet size of the main channel and other elements to increase the main channel flow and optimize the flow ratio between the main channel and the secondary channel, the mixing was improved, which consequently improved the hydraulic performance of the emitter.

As a key component of a drip irrigation system, the performance of drip irrigation emitters is determined by the flow channel structures and parameters. In this paper, a novel Y-shaped labyrinth-channel emitter was proposed, and the influence of the hydraulic performance was researched under different waist-arc angles (α) and ratios of crown height to chord length (a:b). The flow characteristics and energy dissipation mechanisms of water in the emitter were analyzed by using fluid dynamics software. The analysis showed that, keeping the ratio of crown height to chord length unchanged and with an increase in the waist-arc angle from 90° to 180°, the pressure-drop ratio before and after the water flow through the vortex area increased, and the hydraulic performance of the emitter was improved, specifically: the design flow could be reduced by 6.02–26.7%, and the slope of the curve could be reduced by 9.83–28.1%. The smaller the ratio of crown height to chord length, the weaker the influence of waist-arc angle was on the design flow and hydraulic performance. Keeping the waist-arc angle unchanged, there was a decrease in the ratio of crown height to chord length from 6:30 to 1:30, the vortex strength inside the emitter channel increased, and the hydraulic performance was improved: the design flow could be reduced by 7.56–23.5%, and the slope of the curve could be reduced by 5.43–20.5%. The bigger the waist-arc angle, the weaker the influence of the ratio of crown height to chord length was on the design flow and the hydraulic performance, even to the point of having no effect. When the Y-shaped emitter was close to the pressure-flow curve slope of the commonly-used triangular-channel emitter, the hydraulic performance of the two was similar, and the former could increase the channel width by 25%, as compared to the latter, or shorten the channel length by 44.3%. The results showed that the Y-shaped emitter had better hydraulic performance compared with the triangular-flow-channel emitter commonly used today. Therefore, the Y-shaped emitter has broad application prospects in water-saving irrigation.

The opposed-power reciprocating pump has the characteristics of high pressure, large flow, and high efficiency and energy saving. However, due to the special structure of the opposed-power reciprocating pump, existing theoretical methods cannot analyze its dynamic performance. Therefore, this paper proposes a method of analyzing the power end of the opposed-power reciprocating pump. Firstly, according to the working principle and structural characteristics of the traditional plunger pump, the novel and complex structure of the opposed-power reciprocating pump is analyzed by analogy, and the force analysis model of the crankshaft is established. The dynamic analysis model of the Matlab program is used to solve the dynamic load and section stress in the working process, and the variation law of crankshaft load is obtained. The 25 most critical working conditions are selected for analysis, and the most critical station and section of the crankshaft are obtained. With the connection between ANSYS Workbench and Solidworks, the model is imported into ANSYS Workbench, the load on the crank pin is loaded by APDL command flow, and the static analysis of the crankshaft is carried out to obtain the stress and strain of the crankshaft. Finally, the static and fatigue strength of the dangerous section is checked, and it is proven that the strength and stiffness of the crankshaft meet the design requirements. The results show that the dynamic analysis results of the crankshaft under critical working conditions are consistent with the finite element analysis, verifying the rationality of the method and providing a reference for the improvement and optimized design of the crankshaft of the opposed-power reciprocating pump.

In this paper, a split-flow channel layout with one (group) inlet and two (group) outlets is adopted, based on computational fluid dynamics technology, and compared with the current commonly used channel with one (group) inlet and one (group) outlet emitter. On the premise of the same outlet spacing, the pressure–flow relationship curve and slope of the split-flow emitter were analyzed under the three channel layouts of non-return, single-sided re-entry, and bilateral re-entry, with different channel widths and lengths. When exploring the influence of the channel layout and size on the hydraulic performance of split-flow emitters, the results showed that when the split-flow emitter with a non-return channel is adopted and the hydraulic performance is not reduced, the single-side channel length is half that of the one-in-one-out emitter, meaning the channel width needs to be reduced by 15%. When the channel layout is a single-sided channel re-entry, the hydraulic performance is better than that of the one-in-one-out emitter; if the hydraulic performance of the two remains unchanged, the channel width can be increased by 10% or the single-sided channel length can be reduced by 20%. When the channel layout is a bilateral channel re-entry, the channel width can be increased by nearly 30% if the hydraulic performance of the 2 is consistent, and the single-side channel length is increased by about 50%. When the split-flow emitter adopts a non-return channel layout, the channel width needs to be reduced to ensure the hydraulic performance is consistent. If the layout of single-sided channel re-entry or bilateral channel re-entry is adopted, the hydraulic performance is better than that of the one-in-one-out emitter and the hydraulic performance of the two is consistent. Thus, the channel length can be reduced or the channel width increased, which is beneficial for improving the anti-clogging performance of the emitter.

The maximum gasification rate of corn stalk char (CSC) appeared at high conversion range, and its quite different gasification behaviors from other carbonaceous materials are all derived from the catalytic effect of alkali and alkali earth metals (AAEMs), so it is necessary to study the effect of AAEMs and gasification kinetics of such biomass char. However, there are few systematic discussions about this effect and kinetic modeling. Thus, in this study, CSC samples were prepared in a fast pyrolysis fixed-bed reactor, and its gasification experiments were conducted on a pressurized magnetic suspension balance at various total pressures (0.1–0.7 MPa), steam concentrations (10–70 vol.%) and temperatures (725–900 °C). Moreover, a water-leached CSC (H2O-CSC) was also prepared to evaluate the impact of AAEMs on the gasification performance of CSC, and some well-known models were adopted to describe the gasification behaviors. On the basis of these results, the effect of primary AAEMs on the gasification behaviors of CSC and gasification kinetic modeling were obtained. Results showed total pressure had no obvious influence on the gasification rate of CSC, and the reaction order varied at 0.43–0.55 with respect to steam partial pressures. In addition, the modified random pore model (MRPM) and Langmuir–Hinshelwood (L-H) model were satisfactorily applied to predict the gasification behaviors of CSC. The catalytic effect of AAEMs on CSC gasification was weakened due to water-leaching treatment. A random pore model (RPM) could describe the gasification behavior of H2O-CSC well, followed by grain model (GM) and volumetric model (VM).

Total dissolved gas (TDG) supersaturation, which can be caused by flood discharge, results in gas bubble disease (GBD) in fish and threatens their survival downstream of dams. TDG supersaturation has become a serious environmental problem in the Yangtze River. Few studies have evaluated the effect of TDG supersaturation on fish in natural rivers during periods of flood discharge. To estimate fish tolerance to TDG supersaturation under natural conditions, juvenile Myxocyprinus asiaticus and juvenile Procypris rabaudi were exposed to TDG-supersaturated water for 96 h at various depths (0–0.3 m, 0.3–1.3 m, 1.3–2.3 m and 0–2.3 m) during periods of flood discharge of Dagangshan hydropower station. The results showed that juvenile Procypris rabaudi and juvenile Myxocyprinus asiaticus exhibited obvious GBD signs. An increase in exposure time decreased survival probability of the two species. Deeper water depths can increase the tolerance of juvenile Procypris rabaudi to TDG supersaturation in natural rivers during periods of flood discharge while it cannot improve the survival of juvenile Myxocyprinus asiaticus. Compared with juvenile Myxocyprinus asiaticus, juvenile Procypris rabaudi showed weaker tolerance of TDG supersaturation in shallow water, and juvenile Procypris rabaudi were more vulnerable to TDG supersaturation than juvenile Myxocyprinus asiaticus even if the TDG level (116%) was low.

The rectangular labyrinth emitter is taken as the study object in this article, as we added internal teeth to vortex-free and vortex areas in its lateral channel or lengthened the vertical channel, to change the channel structure. Using the computational fluid dynamics (CFD) method simulates the water flow field, to get the relationship between flow rate and pressure, and the vortexes distribution in channel. The aim of this study is to explore the reasons for the influence of structural change on hydraulic performance of the emitter through the analysis of vortex intensity and its distribution from the perspective of the vortex. The results show that the relative error of simulated results and experimental data was 1.02–2.11%. Adding internal teeth to vortex-free areas in lateral channel can improve hydraulic performance of the emitter; adding them to vortex areas can reduce it. The increase in vortex number and intensity in flow field is the internal reason for the improvement of the emitter’s hydraulic performance. The channel structure changes promote the formation of a larger velocity gradient, and the increase in the velocity gradient in flow field exacerbates vortex formation. Changing channel structure to improve the emitter’s hydraulic performance can promote an increase in the number and intensity of vortexes in the channel.

As important transcription factors, WRKYs play a vital role in the defense response of plants against the invasion of multiple pathogens. Though some WRKY members have been reported to participate in pepper immunity in response to Ralstonia solanacearum infection, the functions of the majority of WRKY members are still unknown. Herein, CaWRKY22b was cloned from the pepper genome and its function against R. solanacearum was analyzed. The transcript abundance of CaWRKY22b was significantly increased in response to the infection of R. solanacearum and the application of exogenous methyl jasmonate (MeJA). Subcellular localization assay in the leaves of Nicotiana benthamiana showed that CaWRKY22b protein was targeted to the nuclei. Agrobacterium-mediated transient expression in pepper leaves indicated that CaWRKY22b overexpression triggered intensive hypersensitive response-like cell death, H2O2 accumulation, and the up-regulation of defense- and JA-responsive genes, including CaHIR1, CaPO2, CaBPR1, and CaDEF1. Virus-induced gene silencing assay revealed that knock-down of CaWRKY22b attenuated pepper’s resistance against R. solanacearum and the up-regulation of the tested defense- and jasmonic acid (JA)-responsive genes. We further assessed the role of CaWRKY22b in modulating the expression of JA-responsive CaDEF1, and the result demonstrated that CaWRKY22b trans-activated CaDEF1 expression by directly binding to its upstream promoter. Collectively, our results suggest that CaWRKY22b positively regulated pepper immunity against R. solanacearum in a manner associated with JA signaling, probably by modulating the expression of JA-responsive CaDEF1.

As an important member of mitogen-activated protein kinase (MAPK) cascades, MAPKs play an important role in plant defense response against biotic and abiotic stresses; however, the involvement of the majority of the MAPK family members against Ralstonia solanacearum and heat stress (HS) remains poorly understood. In the present study, CaMAPK1 was identified from the genome of pepper and its function against R. solanacearum and HS was analyzed. The transcript accumulations of CaMAPK1 and the activities of its native promoter were both significantly induced by R. solanacearum inoculation, HS, and the application of exogenous hormones, including SA, MeJA, and ABA. Transient expression of CaMAPK1 showed that CaMAPK1 can be targeted throughout the whole cells in Nicotiana benthamiana and triggered chlorosis and hypersensitive response-like cell death in pepper leaves, accompanied by the accumulation of H2O2, and the up-regulations of hormones- and H2O2-associated marker genes. The knock-down of CaMAPK1 enhanced the susceptibility to R. solanacearum partially by down-regulating the expression of hormones- and H2O2-related genes and impairing the thermotolerance of pepper probably by attenuating CaHSFA2 and CaHSP70-1 transcripts. Taken together, our results revealed that CaMAPK1 is regulated by SA, JA, and ABA signaling and coordinates responses to R. solanacearum infection and HS in pepper.

Lactobacillus plantarum is a kind of probiotic that benefits the host by regulating the gut microbiota, but it is easily damaged when passing through the gastrointestinal tract, hindering its ability to reach the destination and reducing its utilization value. Encapsulation is a promising strategy for solving this problem. In this study, transglutaminase (TGase)-crosslinked gelatin (GE)/sodium hexametaphosphate (SHMP) hydrogels were used to encapsulate L. plantarum. The effects of TGase concentration and drying method on the physiochemical properties of the hydrogels were determined. The results showed that at a TGase concentration of 9 U/gGE, the hardness, chewiness, energy storage modulus, and apparent viscosity of the hydrogel encapsulation system were maximized. This concentration produced more high-energy isopeptide bonds, strengthening the interactions between molecules, forming a more stable three-dimensional network structure. The survival rate under the simulated gastrointestinal conditions and storage stability of L. plantarum were improved at this concentration. The thermal stability of the encapsulation system dried via microwave vacuum freeze drying (MFD) was slightly higher than that when dried via freeze drying (FD). The gel structure was more stable, and the activity of L. plantarum decreased more slowly during the storage period when dried using MFD. This research provides a theoretical basis for the development of encapsulation technology of probiotics.

Sanguinarine (SAN), as the main active component of a traditional Chinese veterinary medicine, has been widely used in the animal husbandry and breeding industry. However, the metabolites of SA are still uncertain. Therefore, this research aimed to investigate the metabolites of SA based on rats in vivo. The blood, feces, and urine of rats were collected after the oral administration of 40 mg/kg SAN. Ultra-high-performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF-MS/MS) was employed to identify the metabolites of SAN. The elemental composition of sanguinarine metabolites was inferred by analyzing their exact molecular weight, and the structures of the metabolites were predicted based on their fragment ions and cleavage pathways. A total of 12 metabolites were identified, including three metabolites in the plasma, four in the urine, and nine in the feces. According to the possible metabolic pathways deduced in this study, SAN was mainly metabolized through reduction, oxidation, demethylation, hydroxylation, and glucuronidation. This present research has summarized the metabolism of SAN in rats, which is helpful for further studying the metabolic mechanism of SAN in vivo and in vitro.

Background: In this study, an apple foam net packing machine is designed to improve the efficiency of apple foam net packing, reduce the intensity of manual work, and meet the market demand for diverse forms of apple packing. Methods: Based on the analysis of apple shape characteristics, a secondary posture adjustment method for apples with a vertical posture adjustment mechanism and a side posture adjustment mechanism is proposed. Based on the observation of human hand-set nets and the analysis of the mechanical characteristics of nets, a dual-system net opening method combining a net slackening system and a net opening system is proposed. Results: The experiment shows that: the apple foam net sleeve packing machine can reach 99% of the apple’s posture adjustment rate, the net opening rate can reach 99%, and the packing rate can reach 98.10%. The packing speed is 10 s/pcs, and the packing efficiency is 355–365 pcs/h. The length of the fused mesh net sleeve is mainly distributed between 138 and 143 mm, which accounts for approximately 80% of the total number of samples. Conclusions: The apples packed by this machine are neatly packaged according to specifications. It can improve the efficiency of apple foam net sleeve packing and replace manual work.

Conventional reservoir drill-in fluids used for drilling reservoirs in Weizhou Oilfield encounter rheological problems that result in technical problems such as hole-cleaning in openhole horizontal intervals. Hence, novel drill-in fluid was developed by optimizing the additive quantity and particle size distribution. Lab tests showed that novel drill-in fluids boast high low shearing rate viscosity, and provide promising cutting, carrying, and suspension capabilities. Furthermore, the novel drill-in fluids performed well in reservoir protection, with a permeability recovery rate of more than 90%. The novel drill-in fluids also have high inhibition capabilities with a linear expansion rate of mud shale as low as 10%, with a rolling recovery rate of up to 96.48%. Field application results showed no pipe-stuck was encountered during tripping in the horizontal interval when using the novel drill-in fluid. Moreover, by using the novel drill-in fluids, skin factor was reduced from 20.0 to −3.0, and daily oil production was double what was expected. It was concluded that novel drill-in fluids meets the demand of horizontal drilling intervals in Weizhou Oilfield and improves hole-cleaning and reservoir protection in the horizontal well.

Proteins with conserved SET domain play a critical role in plant immunity. However, the means of organization and functions of these proteins are unclear, particularly in non-model plants such as pepper (Capsicum annum L.). Herein, we functionally characterized CaASHH3, a member of class II (the ASH1 homologs H3K36) proteins in pepper immunity against Ralstonia solanacearum and Pseudomonas syringae pv tomato DC3000 (Pst DC3000). The CaASHH3 was localized in the nucleus, and its transcript levels were significantly enhanced by R. solanacearum inoculation (RSI) and exogenous application of salicylic acid (SA), methyl jasmonate (MeJA), ethephon (ETH), and abscisic acid (ABA). Knockdown of CaASHH3 by virus-induced gene silencing (VIGS) compromised peppers’ resistance to RSI. Furthermore, silencing of CaASHH3 impaired hypersensitive-response (HR)-like cell death response due to RSI and downregulated defense-associated marker genes, including CaPR1, CaNPR1, and CaABR1. The CaASHH3 protein was revealed to affect the promoters of CaNPR1, CaPR1, and CaHSP24. Transiently over-expression of CaASHH3 in pepper leaves elicited HR-like cell death and upregulated immunity-related marker genes. To further study the role of CaASHH3 in plant defense in vivo, CaASHH3 transgenic plants were generated in Arabidopsis. Overexpression of CaASHH3 in transgenic Arabidopsis thaliana enhanced innate immunity against Pst DC3000. Furthermore, CaASHH3 over-expressing transgenic A. thaliana plants exhibited upregulated transcriptional levels of immunity-associated marker genes, such as AtNPR1, AtPR1, and AtPR2. These results collectively confirm the role of CaASHH3 as a positive regulator of plant cell death and pepper immunity against bacterial pathogens, which is regulated by signaling synergistically mediated by SA, JA, ET, and ABA.

The WRKY transcription factors (TFs) network is composed of WRKY TFs’ subset, which performs a critical role in immunity regulation of plants. However, functions of WRKY TFs’ network remain unclear, particularly in non-model plants such as pepper (Capsicum annuum L.). This study functionally characterized CaWRKY30—a member of group III Pepper WRKY protein—for immunity of pepper against Ralstonia solanacearum infection. The CaWRKY30 was detected in nucleus, and its transcriptional expression levels were significantly upregulated by R. solanacearum inoculation (RSI), and foliar application ethylene (ET), abscisic acid (ABA), and salicylic acid (SA). Virus induced gene silencing (VIGS) of CaWRKY30 amplified pepper’s vulnerability to RSI. Additionally, the silencing of CaWRKY30 by VIGS compromised HR-like cell death triggered by RSI and downregulated defense-associated marker genes, like CaPR1, CaNPR1, CaDEF1, CaABR1, CaHIR1, and CaWRKY40. Conversely, transient over-expression of CaWRKY30 in pepper leaves instigated HR-like cell death and upregulated defense-related maker genes. Furthermore, transient over-expression of CaWRKY30 upregulated transcriptional levels of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40. On the other hand, transient over-expression of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40 upregulated transcriptional expression levels of CaWRKY30. The results recommend that newly characterized CaWRKY30 positively regulates pepper’s immunity against Ralstonia attack, which is governed by synergistically mediated signaling by phytohormones like ET, ABA, and SA, and transcriptionally assimilating into WRKY TFs networks, consisting of CaWRKY6, CaWRKY22, CaWRKY27, and CaWRKY40. Collectively, our data will facilitate to explicate the underlying mechanism of crosstalk between pepper’s immunity and response to RSI.

‘Candidatus Liberibacter asiaticus’ (CLas) is the pathogenic bacterium that causes the disease Huanglongbing (HLB) in citrus and some model plants, such as Nicotiana benthamiana. After infection, CLas releases a set of effectors to modulate host responses. One of these critical effectors is Sec-delivered effector 1 (SDE1), which induces chlorosis and cell death in N. benthamiana. In this study, we revealed the DEAD-box RNA helicase (DDX3) interacts with SDE1. Gene silencing study revealed that knockdown of the NbDDX3 gene triggers leaf chlorosis, mimicking the primary symptom of CLas infection in N. benthamiana. The interactions between SDE1 and NbDDX3 were localized in the cell membrane. Overexpression of SDE1 resulted in suppression of NbDDX3 gene expression in N. benthamiana, which suggests a critical role of SDE1 in modulating NbDDX3 expression. Furthermore, we verified the interaction of SDE1 with citrus DDX3 (CsDDX3), and demonstrated that the expression of the CsDDX3 gene was significantly reduced in HLB-affected yellowing and mottled leaves of citrus. Thus, we provide molecular evidence that the downregulation of the host DDX3 gene is a crucial mechanism of leaf chlorosis in HLB-affected plants. The identification of CsDDX3 as a critical target of SDE1 and its association with HLB symptom development indicates that the DDX3 gene is an important target for gene editing, to interrupt the interaction between DDX3 and SDE1, and therefore interfere host susceptibility.

In this paper, Nd3+–Yb3+–Er3+-doped β-NaYF4 nanocrystals with different Nd3+ concentrations are synthesized, and the luminescence properties of the upconversion nanoparticles (UCNPs) have been studied under 808-nm excitation for sensitive biological applications. The upconversion luminescence spectra of NaYF4 nanoparticles with different dopants under 808-nm excitation proves that the Nd3+ ion can absorb the photons effectively, and the Yb3+ ion can play the role of an energy-transfer bridging ion between the Nd3+ ion and Er3+ ion. To investigate the effect of the Nd3+ ion, the decay curves of the 4S3/2 → 4I15/2 transition at 540 nm are measured and analyzed. The NaYF4: 20% Yb3+, 2% Er3+, 0.5% Nd3+ nanocrystals have the highest emission intensity among all samples under 808-nm excitation. The UC (upconversion) mechanism under 808-nm excitation is discussed in terms of the experimental results.

GSK3-like kinases have been mainly implicated in the brassinosteroids (BR) pathway and, therefore, in plant growth, development, and responses to abiotic stresses; however, their roles in plant immunity remain poorly understood. Herein, we present evidence that CaSK23, a putative GSK3/SHAGGY-like kinase in pepper, acts as a negative regulator in pepper’s response to Ralstonia solanacearum (R. solanacearum) inoculation (RSI). Data from quantitative RT-PCR (qRT-PCR) showed that the constitutively-expressed CaSK23 in pepper leaves was down-regulated by RSI, as well as by exogenously-applied salicylic acid (SA) or methyl jasomonate (MeJA). Silencing of CaSK23 by virus-induced gene silencing (VIGS) decreased the susceptibility of pepper plants to RSI, coupled with up-regulation of the tested genes encoding SA-, JA-, and ethylene (ET)-dependent pathogenesis-related (PR) proteins. In contrast, ectopic overexpression (OE) of CaSK23 conferred a compromised resistance of tobacco plants to RSI, accompanied by down-regulation of the tested immunity-associated SA-, JA-, and ET-dependent PR genes. In addition, transient overexpression of CaSK23 in pepper plants consistently led to down-regulation of the tested SA-, JA-, and ET-dependent PR genes. We speculate that CaSK23 acts as a negative regulator in pepper immunity and its constitutive expression represses pepper immunity in the absence of pathogens. On the other hand, its decreased expression derepresses immunity when pepper plants are attacked by pathogens.

WRKY transcription factors (TFs) have been implicated in plant growth, development, and in response to environmental cues; however, the function of the majority of pepper WRKY TFs remains unclear. In the present study, we functionally characterized CaWRKY40b, a homolog of AtWRKY40, in pepper immunity. Ralstonia solanacearum inoculation (RSI) in pepper plants resulted in downregulation of CaWRKY40b transcript, and green fluorescent protein (GFP)-tagged CaWRKY40b was localized to the nuclei when transiently overexpressed in the leaves of Nicotiana benthamiana. Virus-induced gene silencing (VIGS) of CaWRKY40b significantly decreased pepper’ susceptibility to RSI. Consistently, the transient over-expression of CaWRKY40b-SRDX (chimeric repressor version of CaWRKY40b) triggered cell death, as indicated by darker trypan blue and DAB staining. CaWRKY40b targets a number of immunity-associated genes, including CaWRKY40 JAR, RLK1, EIN3, FLS2, CNGIC8, CDPK13, and heat shock cognate protein 70 (HSC70), which were identified by ChIP-seq and confirmed using ChIP-real time PCR. Among these target genes, the negative regulator HSC70 was upregulated by transient overexpression of CaWRKY40b and downregulated by silencing of CaWRKY40b, whereas other positive regulators as well as two non-target genes, CaNPR1 and CaDEF1, were downregulated by the transient overexpression of CaWRKY40b and upregulated by CaWRKY40b silencing or transient overexpression of CaWRKY40b-SRDX. In addition, CaWRKY40b exhibited a positive feedback regulation at transcriptional level by directly targeting the promoter of itself. In conclusion, the findings of the present study suggest that CaWRKY40b acts as a negative regulator in pepper immunity against R. solanacearum by transcriptional modulation of a subset of immunity-associated genes; it also represses immunity in the absence of a pathogen, and derepresses immunity upon pathogen challenge.

The WRKY web, which is comprised of a subset of WRKY transcription factors (TFs), plays a crucial role in the regulation of plant immunity, however, the mode of organization and operation of this network remains obscure, especially in non-model plants such as pepper (Capsicum annuum). Herein, CaWRKY22, a member of a subgroup of IIe WRKY proteins from pepper, was functionally characterized in pepper immunity against Ralstonia Solanacearum. CaWRKY22 was found to target the nuclei, and its transcript level was significantly upregulated by Ralstonia Solanacearum inoculation (RSI) and exogenously applied salicylic acid (SA), Methyl jasmonate (MeJA), or ethephon (ETH). Loss-of-function CaWRKY22, caused by virus-induced gene silencing (VIGS), enhanced pepper’s susceptibility to RSI. In addition, the silencing of CaWRKY22 perturbed the hypersensitive response (HR)-like cell death elicited by RSI and downregulated defense-related genes including CaPO2, CaPR4, CaACC, CaBPR1, CaDEF1, CaHIR1, and CaWRKY40. CaWRKY22 was found to directly bind to the promoters of CaPR1, CaDEF1, and CaWRKY40 by chromatin immuno-precipitation (ChIP) analysis. Contrastingly, transient overexpression of CaWRKY22 in pepper leaves triggered significant HR-like cell death and upregulated the tested immunity associated maker genes. Moreover, the transient overexpression of CaWRKY22 upregulated the expression of CaWRKY6 and CaWRKY27 while it downregulated of the expression of CaWRKY58. Conversely, the transient overexpression of CaWRKY6, CaWRKY27, and CaWRKY40 upregulated the expression of CaWRKY22, while transient overexpression of CaWRKY58 downregulated the transcript levels of CaWRKY22. These data collectively recommend the role of CaWRKY22 as a positive regulator of pepper immunity against R. Solanacearum, which is regulated by signaling synergistically mediated by SA, jasmonic acid (JA), and ethylene (ET), integrating into WRKY networks with WRKY TFs including CaWRKY6, CaWRKY27, CaWRKY40, and CaWRKY58.

In this study, 27 polarimetric parameters were extracted from Radarsat-2 polarimetric synthetic aperture radar (SAR) at each growth stage of the rape crop. The sensitivity to growth parameters such as stem height, leaf area index (LAI), and biomass were investigated as a function of days after sowing. Based on the sensitivity analysis, five empirical regression models were compared to determine the best model for stem height, LAI, and biomass inversion. Of these five models, quadratic models had higher R2 values than other models in most cases of growth parameter inversions, but when these results were related to physical scattering mechanisms, the inversion results produced overestimation in the performance of some parameters. By contrast, linear and logarithmic models, which had lower R2 values than the quadratic models, had stable performance for growth parameter inversions, particularly in terms of their performance at each growth stage. The best biomass inversion performance was acquired by the volume component of a quadratic model, with an R2 value of 0.854 and root mean square error (RMSE) of 109.93 g m−2. The best LAI inversion was also acquired by a quadratic model, but used the radar vegetation index (Cloude), with an R2 value of 0.8706 and RMSE of 0.56 m2 m−2. Stem height was acquired by scattering angle alpha ( α ) using a logarithmic model, with an R2 of 0.926 value and RMSE of 11.09 cm. The performances of these models were also analysed for biomass estimation at the second growth stage (P2), third growth stage (P3), and fourth growth stage (P4). The results showed that the models built at the P3 stage had better substitutability with the models built during all of the growth stages. From the mapping results, we conclude that a model built at the P3 stage can be used for rape biomass inversion, with 90% of estimation errors being less than 100 g m−2.

Despite the involvement of many members of the chitinase family in plant immunity, the precise functions of the majority of the members remain poorly understood. Herein, the gene ChiIV3 in Capsicum annuum encoding a chitinase protein containing a chitin binding domain and targeting to the plasma membrane was found to be induced by Phytophthora capsici inoculation (PCI) and applied chitin treatment. Besides its direct inhibitory effect on growth of Phytophthora capsici (P. capsici), ChiIV3 was also found by virus-induced gene silencing (VIGS) and transient overexpression (TOE) in pepper plants to act as a positive regulator of plant cell death and in triggering defense signaling and upregulation of PR (pathogenesis related) genes against PCI. A 5′ deletion assay revealed that pChiIV3−712 to −459 bp was found to be sufficient for ChiIV3’ response to PCI. Furthermore, a mutation assay indicated that W-box−466 to −461 bp in pChiIV3−712 to −459 bp was noted to be the PCI-responsible element. These results collectively suggest that ChiIV3 acts as a likely antifungal protein and as a receptor for unidentified chitin in planta to trigger cell death and defense signaling against PCI.