A comprehensive analysis of the WRKY family in soybean and functional analysis of GmWRKY164-GmGSL7c in resistance to soybean mosaic virus.

BACKGROUND: Soybean mosaic disease caused by soybean mosaic virus (SMV) is one of the most devastating and widespread diseases in soybean producing areas worldwide. The WRKY transcription factors (TFs) are widely involved in plant development and stress responses. However, the roles of the GmWRKY TFs in resistance to SMV are largely unclear. RESULTS: Here, 185 GmWRKYs were characterized in soybean (Glycine max), among which 60 GmWRKY genes were differentially expressed during SMV infection according to the transcriptome data. The transcriptome data and RT-qPCR results showed that the expression of GmWRKY164 decreased after imidazole treatment and had higher expression levels in the incompatible combination between soybean cultivar variety Jidou 7 and SMV strain N3. Remarkably, the silencing of GmWRKY164 reduced callose deposition and enhanced virus spread during SMV infection. In addition, the transcript levels of the GmGSL7c were dramatically lower upon the silencing of GmWRKY164. Furthermore, EMSA and ChIP-qPCR revealed that GmWRKY164 can directly bind to the promoter of GmGSL7c, which contains the W-box element. CONCLUSION: Our findings suggest that GmWRKY164 plays a positive role in resistance to SMV infection by regulating the expression of GmGSL7c, resulting in the deposition of callose and the inhibition of viral movement, which provides guidance for future studies in understanding virus-resistance mechanisms in soybean.

Deciphering the crop-soil-enzyme C:N:P stoichiometry nexus: A 5-year study on manure-induced changes in soil phosphorus transformation and release risk.

Carbon:nitrogen:phosphorus (C:N:P) stoichiometry plays a vital role in regulating P transformation in agriculture ecosystems. However, the impact of balanced C:N:P stoichiometry in paddy soil, particularly regarding relative soil P transformation, remains unknown. This study explores the response of C:N:P stoichiometry to manure substitution and its regulatory role in soil P transformation, along with the associated release risk to the environment. Based on a 5-year field study, our findings reveal that replacing 30 % of chemical P fertilizer with pig manure (equal total NPK amounts with chemical P fertilizer treatment, named CFM) increased soil total C without altering soil total P, resulting in an elevated soil C:P ratio, despite the homeostasis of crop stoichiometry. This increase promoted microbial diversity and the accumulation of organic P in the soil. The Proteobacteria and Actinobacteria produced lower C:PEEA metabolism together, and enhanced in vivo turnover of P. Additionally, by integrating high-resolution dialysis (HR-Peeper), diffusive gradients in thin films (DGT), DGT-induced fluxes in the soil (DIFS), and sediment P release risk index (SPRRI) models, we observed that, in addition to organic P, CFM simultaneously increased soil Al-P, thereby weakening the diffusion and resupply capacity of P from soil solids to the solution. Consequently, this decrease in P release risk to the environment was demonstrated. Overall, this study establishes a connection between crop-soil-enzyme C:N:P stoichiometry, soil microorganisms, and soil P biogeochemical processes. The study further evaluates the P release risk to the environment, providing a novel perspective on both the direct and indirect effects of manure substitution on soil P cycling.

Are digital technologies an effective inhibitor of depression among middle-aged and older adults? Micro-level evidence from a panel study.

The increased number of middle-aged and older adults leads to depression in this stratum of the population as a topical social and public health issue. However, the new generation of information technologies has exerted a profound impact on the lives of middle-aged and older adults, and offers potential solutions for alleviating their depression. This study utilizes data from the China Health and Retirement Longitudinal Study (CHARLS) collected between 2011 and 2018 and combines them with city-level traits. The results demonstrate that digital technology can reduce depression levels effectively in this group. Mechanism analysis reveals that digital technology could improve life satisfaction and subjective health status levels, which, in turn, reduces depression levels. Heterogeneity analysis shows that the positive effects of digital technology on depression were more pronounced among middle-aged and older adults with urban household registration compared to the rural population. Finally, recommendations are provided for reducing depression levels among middle-aged and older adults.

CL-SPO2Net: Contrastive Learning Spatiotemporal Attention Network for Non-Contact Video-Based SpO2 Estimation.

Video-based peripheral oxygen saturation (SpO2) estimation, utilizing solely RGB cameras, offers a non-contact approach to measuring blood oxygen levels. Previous studies set a stable and unchanging environment as the premise for non-contact blood oxygen estimation. Additionally, they utilized a small amount of labeled data for system training and learning. However, it is challenging to train optimal model parameters with a small dataset. The accuracy of blood oxygen detection is easily affected by ambient light and subject movement. To address these issues, this paper proposes a contrastive learning spatiotemporal attention network (CL-SPO2Net), an innovative semi-supervised network for video-based SpO2 estimation. Spatiotemporal similarities in remote photoplethysmography (rPPG) signals were found in video segments containing facial or hand regions. Subsequently, integrating deep neural networks with machine learning expertise enabled the estimation of SpO2. The method had good feasibility in the case of small-scale labeled datasets, with the mean absolute error between the camera and the reference pulse oximeter of 0.85% in the stable environment, 1.13% with lighting fluctuations, and 1.20% in the facial rotation situation.

Expression and significance of silent information regulator two homolog 1 in the placenta and plasma of patients with pre-eclampsia-a meta-analysis.

OBJECTIVE: This study aimed to collect, organize, and conduct a meta-analysis of the literature on the expression of silent information regulator two homolog 1 (SIRT1) in the placental tissue and plasma of patients with pre-eclampsia. METHODS: The enrolled patients were divided into two groups: the pre-eclampsia group and the healthy group. This study summarized and analyzed the demographic characteristics of the two groups, including pregnancy age, gestational weeks, parity, gravidity, blood pressure, Body Mass Index, newborn weight, placental weight, and SIRT1 expression in placental tissue and maternal plasma. RESULTS: Eleven studies were included in this research, with 586 cases in the pre-eclampsia group and 479 cases in the control group. Three research studies are reporting immunohistochemistry tests, among which the pre-eclampsia group had a positivity rate of 30.24% (62/205), while the control group had 58.02% (76/131); the two groups have a significant difference (p < 0.05). Two research studies reported the results of ELISA tests, with 107 cases in the pre-eclampsia group and 125 cases in the control group. A comparison of the SIRT1 test results showed a statistically significant difference between the two groups (p < 0.05). Pre-eclampsia group patients had lower gestational weeks, newborn birth weight, and placental weight compared to the healthy control group (all p < 0.05). However, systolic and diastolic blood pressures were higher in the pre-eclampsia group than in the control group (p < 0.05). CONCLUSION: SIRT1 expression is downregulated in pre-eclampsia patients' plasma and placental tissue. Further research is needed to validate this conclusion.

A Combined Weighting Based Large Scale Group Decision Making Framework for MOOC Group Recommendation.

Massive open online courses (MOOC) are free learning courses based on online platforms for higher education, which not only promote the open sharing of learning resources, but also lead to serious information overload. However, there are many courses on MOOCs, and it can be difficult for users to choose courses that match their individual or group preferences. Therefore, a combined weighting based large-scale group decision-making approach is proposed to implement MOOC group recommendations. First, based on the MOOC operation mode, we decompose the course content into three stages, namely pre-class, in-class, and post-class, and then the curriculum-arrangement-movement- performance evaluation framework is constructed. Second, the probabilistic linguistic criteria importance through intercriteria correlation method is employed to obtain the objective weighting of the criterion. Meanwhile, the word embedding model is utilized to vectorize online reviews, and the subjective weighting of the criteria are acquired by calculating the text similarity. The combined weighting then can be obtained by fusing the subjective and objective weighting. Based on this, the PL-MULTIMIIRA approach and Borda rule is employed to rank the alternatives for group recommendation, and an easy-to-use formula for group satisfaction is proposed to evaluate the effect of the proposed method. Furthermore, a case study is conducted to group recommendations for statistical MOOCs. Finally, the robustness and effectiveness of the proposed approach were verified through sensitivity analysis as well as comparative analysis.

Segmentation of biventricle in cardiac cine MRI via nested capsule dense network.

Background: Cardiac magnetic resonance image (MRI) has been widely used in diagnosis of cardiovascular diseases because of its noninvasive nature and high image quality. The evaluation standard of physiological indexes in cardiac diagnosis is essentially the accuracy of segmentation of left ventricle (LV) and right ventricle (RV) in cardiac MRI. The traditional symmetric single codec network structure such as U-Net tends to expand the number of channels to make up for lost information that results in the network looking cumbersome. Methods: Instead of a single codec, we propose a multiple codecs structure based on the FC-DenseNet (FCD) model and capsule convolution-capsule deconvolution, named Nested Capsule Dense Network (NCDN). NCDN uses multiple codecs to achieve multi-resolution, which makes it possible to save more spatial information and improve the robustness of the model. Results: The proposed model is tested on three datasets that include the York University Cardiac MRI dataset, Automated Cardiac Diagnosis Challenge (ACDC-2017), and the local dataset. The results show that the proposed NCDN outperforms most methods. In particular, we achieved nearly the most advanced accuracy performance in the ACDC-2017 segmentation challenge. This means that our method is a reliable segmentation method, which is conducive to the application of deep learning-based segmentation methods in the field of medical image segmentation.

PBP1 plays key roles in sex pheromone reception of the fall armyworm.

Spodoptera frugiperda among the China population employs a four-component sex pheromone blend to accelerate male-female allocation and mating behavior. The underlying molecular mechanism has been incompletely elucidated. In the current study, we showed that differences existed between genders toward the four sex pheromone components, including Z9-14:AC, Z7-12:AC, Z9-12:AC, and Z11-16:AC, in terms of electrophysiological responses and behavioral valences. Male adults were significantly more sensitive to all tested compounds than female adults. Furthermore, ecological outputs may be related to four pheromone-binding proteins, namely, SfruPBP1, SfruPBP2, SfruPBP3, and SfruPBP4. They formed four distinct clades within the lepidopteran phylogeny, and male adults expressed significantly higher levels of SfruPBP1 and SfruPBP2 than female adults. We observed the highest binding affinities of SfruPBP1 toward all four sex pheromone components. SfruPBP4 had moderate binding affinities for Z7-12:AC, Z11-16:AC, and Z9-12:AC, while SfruPBP2 showed binding toward Z9-14:AC. This observation suggests that SfruPBP1 plays a key role in sex pheromone discrimination and drives sexually biased behavioral decisions toward certain pheromone components. These findings will help to develop behavioral-mediating tools as part of integrated pest management approaches for this cross-border pest.

Transcriptome Analysis of Detoxification-Related Genes in Spodoptera frugiperda (Lepidoptera: Noctuidae).

Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is an important pest on maize, and it can cause large yield losses. As S. frugiperda has invaded many developing countries in Africa and Asia in recent years, it could impact food security. Pesticides remain the main method to control S. frugiperda in the field, and this pest has developed resistance to some pesticides. In this study, we used second-generation sequencing technology to detect the gene expression change of S. frugiperda after treatment by LC20 of three pesticides, lufenuron, spinetoram, and tetrachloroamide, which have different modes of actions. The sequence data were first assembled into a 60,236 unigenes database, and then the differential expression unigenes (DEUs) after pesticide treatment were identified. The DEU numbers, Gene Ontology catalog, and Kyoto Encyclopedia of Genes and Genomes pathway catalog were analyzed. Finally, 11 types of unigenes related to detoxification and DEUs after pesticide treatment were listed, and Cytochrome P450, Glutathione S-transferase, and ATP-binding cassette transporter were analyzed. This study provides a foundation for molecular research on S. frugiperda pesticide detoxification.

Genome-wide identification and expression analysis of the coronatine-insensitive 1 (COI1) gene family in response to biotic and abiotic stresses in Saccharum.

BACKGROUND: The coronatine insensitive 1 (COI1) gene is the core member of jasmonate signaling pathway, which is closely related to plant biotic and abiotic resistance. However, there have been no reports on COI1 in sugarcane (Sacharum spp.). Hence, systematically investigating the characteristics of the COI1 multigene family in sugarcane can provide a means to study and manipulate the jasmonic acid signaling pathway. RESULTS: A total of 156 COI1 proteins were obtained from the genomes of 19 land plants, while none were obtained from five algae species. A phylogenetic tree demonstrated that these COI1 proteins were classified into four groups, while 31 proteins of SsCOI1 from Saccharum spontaneum, SbCOI1 from Sorghum bicolor, and ShCOI1 from Saccharum spp. hybrid cultivar R570 clustered into three groups. Synteny analysis and duplication patterns revealed that COI1 genes expanded through various genome replication events and could have experienced strong purifying selective pressure during evolution in S. spontaneum, S. bicolor, and R570. An investigation of cis-acting elements suggests that COI1 genes may be involved in plant growth and development and response to various stresses. Expression analysis implied that 21 SsCOI1 genes were constitutively expressed, and had positive responses to drought, cold, and Sporisorium scitamineum stresses with different expression patterns. Among them, seven SsCOI1 haplotype genes may play different roles in response to methyl jasmonate. Furthermore, the ShCOI1-4, ShCOI1-5, and ShCOI1-6 genes were cloned from Saccharum spp. hybrid cultivar ROC22. Real-time quantitative PCR (RT-qPCR) analysis demonstrated that these three ShCOI1 genes had divergent expression profiles in response to salicylic acid, abscisic acid, polyethylene glycol, cold, and S. scitamineum. CONCLUSIONS: These results suggest that COI1 genes may act in sugarcane growth, development, and response to various stresses via different regulatory mechanisms, which laying a foundation for the functional identification of the sugarcane COI1 gene.

Transcriptome analysis of Holotrichia oblita reveals differentially expressed unigenes related to reproduction and development under different photoperiods.

Holotrichia oblita (Faldermann) (Coleoptera: Scarabaeidae) is an insect whose feeding and mating behaviors occur at night. A scotophase is necessary for H. oblita reproduction. We used RNA sequencing (RNA-seq) to compare the expression patterns of H. oblita at five photoperiods (0:24, 8:16, 12:12, 16:8, and 24:0 h) (L:D). Compared to the control (24:0) (L:D), 161-684 differentially expressed unigenes (DEUs) were found in female samples, while 698-2322 DEUs were found in male samples. For all DEUs, a total of 92-1143 DEUs were allocated to 116-662 categories of gene ontology (GO), and 81-1116 DEUs were assigned into 77-286 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. The iPath diagram showed that the DEUs generated by comparing female and male samples with photoperiods of 0:24 and 24:0, respectively, involved multiple metabolic pathways, such as carbohydrate metabolism, lipid metabolism, nucleotide metabolism, purine metabolism and glutathione metabolism. Most of these DEUs were upregulated. Finally, 13 DEUs related to reproduction and development were selected to confirm the consistency of relative expression between RNA-Seq and quantitative real-time polymerase chain reaction (qRT-PCR). Most of these comparison results agreed well, except for some qRT-PCR results that were not detected in male samples due to their low expression. These results provide useful information for understanding the dark-induced reproduction of H. oblita.

A sugarcane smut fungus effector simulates the host endogenous elicitor peptide to suppress plant immunity.

The smut fungus Sporisorium scitamineum causes the most prevalent disease on sugarcane. The mechanism of its pathogenesis, especially the functions and host targets of its effector proteins, are unknown. In order to identify putative effectors involving in S. scitamineum infection, a weighted gene co-expression network analysis was conducted based on the transcriptome profiles of both smut fungus and sugarcane using a customized microarray. A smut effector gene, termed SsPele1, showed strong co-expression with sugarcane PLANT ELICITOR PEPTIDE RECEPTOR1 (ScPEPR1), which encodes a receptor like kinase for perception of plant elicitor peptide1 (ScPep1). The relationship between SsPele1 and ScPEPR1, and the biological function of SsPele1 were characterized in this study. The SsPele1 C-terminus contains a plant elicitor peptide-like motif, by which SsPele1 interacts strongly with ScPEPR1. Strikingly, the perception of ScPep1 on ScPEPR1 is competed by SsPele1 association, leading to the suppression of ScPEPR1-mediated immune responses. Moreover, the Ustilago maydis effector UmPele1, an ortholog of SsPele1, promotes fungal virulence using the same strategy. This study reveals a novel strategy by which a fungal effector can mimic the plant elicitor peptide to complete its perception and attenuate receptor-activated immunity.

The challenges of COVID-19 control policies for sustainable development of business: Evidence from service industries.

Such large-scale disruptions as the pandemic increase the uncertainty and risk related to business. Therefore, the business continuity management (BCM) has become an essential technical solution for enterprise emergency response. Since the beginning of 2020, the COVID-19 has spread worldwide at an alarming rate causing many threats to sustainable development of the business sector. The decline in consumer demand has hugely impacted service industries, such as wholesale and retail sales, tourism. Enterprise production and operations have faced severe challenges. In this study, we develop a risk factor analysis of BCM under the presence of COVID-19 in China. Based on a statistical survey of 940 enterprises in Hangzhou City, China, this study employs ordinal logistic regression to explore the hindering effect of risk factors introduced by the epidemic on business performance. Then, the interpretive structure model (ISM) is applied to analyze the hierarchical structure of the factors under examination. The key factors influencing the enterprise production and operation during COVID-19 outbreak significantly differ across the sub-sectors of the service industry. Therefore, this paper assesses the resilience of the productive technologies and business models of different industries amid the pandemic. This paper proposes epidemic prevention and control strategy focusing on investment and government regulation to ensure sustainable business development.

A PIP-mediated osmotic stress signaling cascade plays a positive role in the salt tolerance of sugarcane.

BACKGROUND: Plasma membrane intrinsic proteins (PIPs) are plant channel proteins involved in water deficit and salinity tolerance. PIPs play a major role in plant cell water balance and responses to salt stress. Although sugarcane is prone to high salt stress, there is no report on PIPs in sugarcane. RESULTS: In the present study, eight PIP family genes, termed ScPIP1-1, ScPIP1-2, ScPIP1-3, ScPIP1-4, ScPIP2-1, ScPIP2-2, ScPIP2-4 and ScPIP2-5, were obtained based on the sugarcane transcriptome database. Then, ScPIP2-1 in sugarcane was cloned and characterized. Confocal microscopy observation indicated that ScPIP2-1 was located in the plasma membrane and cytoplasm. A yeast two-hybridization experiment revealed that ScPIP2-1 does not have transcriptional activity. Real time quantitative PCR (RT-qPCR) analysis showed that ScPIP2-1 was mainly expressed in the leaf, root and bud, and its expression levels in both below- and aboveground tissues of ROC22 were up-regulated by abscisic acid (ABA), polyethylene glycol (PEG) 6000 and sodium chloride (NaCl) stresses. The chlorophyll content and ion leakage measurement suggested that ScPIP2-1 played a significant role in salt stress resistance in Nicotiana benthamiana through the transient expression test. Overexpression of ScPIP2-1 in Arabidopsis thaliana proved that this gene enhanced the salt tolerance of transgenic plants at the phenotypic (healthier state, more stable relative water content and longer root length), physiologic (more stable ion leakage, lower malondialdehyde content, higher proline content and superoxide dismutase activity) and molecular levels (higher expression levels of AtKIN2, AtP5CS1, AtP5CS2, AtDREB2, AtRD29A, AtNHX1, AtSOS1 and AtHKT1 genes and a lower expression level of the AtTRX5 gene). CONCLUSIONS: This study revealed that the ScPIP2-1-mediated osmotic stress signaling cascade played a positive role in plant response to salt stress.

The CaCA superfamily genes in Saccharum: comparative analysis and their functional implications in response to biotic and abiotic stress.

BACKGROUND: In plants, Calcium (Ca2+) acts as a universal messenger in various signal transduction pathways, including responses to biotic and abiotic stresses and regulation of cellular and developmental processes. The Ca2+/cation antiporter (CaCA) superfamily proteins play vital roles in the transport of Ca2+ and/or other cations. However, the characteristics of these superfamily members in Saccharum and their evolutionary and functional implications have remained unclear. RESULTS: A total of 34 CaCA genes in Saccharum spontaneum, 5 CaCA genes in Saccharum spp. R570, and 14 CaCA genes in Sorghum bicolor were identified and characterized. These genes consisted of the H+/cation exchanger (CAX), cation/Ca2+ exchanger (CCX), EF-hand / CAX (EFCAX), and Mg2+/H+ exchanger (MHX) families, among which the CCX and EFCAX could be classified into three groups while the CAX could be divided into two groups. The exon/intron structures and motif compositions suggested that the members in the same group were highly conserved. Synteny analysis of CaCAs established their orthologous and paralogous relationships among the superfamily in S. spontaneum, R570, and S. bicolor. The results of protein-protein interactions indicated that these CaCA proteins had direct or indirect interactions. Quantitative reverse transcription polymerase chain reaction (qRT-PCR) analysis demonstrated that most members of Saccharum CaCA genes exhibited a similar expression pattern in response to hormonal (abscisic acid, ABA) treatment but played various roles in response to biotic (Sporisorium scitamineum) and abiotic (cold) stresses. Furthermore, ScCAX4, a gene encoding a cytoplasm, plasma membrane and nucleus positioning protein, was isolated from sugarcane. This gene was constitutively expressed in different sugarcane tissues and its expression was only induced at 3 and 6 h time points after ABA treatment, however was inhibited and indued in the whole process under cold and S. scitamineum stresses, respectively. CONCLUSIONS: This study systematically conducted comparative analyses of CaCA superfamily genes among S. spontaneum, R570, and S. bicolor, delineating their sequence and structure characteristics, classification, evolutionary history, and putative functions. These results not only provided rich gene resources for exploring the molecular mechanism of the CaCA superfamily genes but also offered guidance and reference for research on other gene families in Saccharum.

Effect of Photoperiod on Longevity, Food Consumption, and Reproduction of Holotrichia oblita (Coleoptera: Scarabaeidae).

Holotrichia oblita (Faldermann) (Coleoptera: Scarabaeidae) is a major soil insect pest that damages forest trees, crops, and lawns. Adults of H. oblita fly, forage, and mate at night but remain underground during the day. We studied the effect of photoperiod on H. oblita reproduction. H. oblita females laid more eggs at 8:16 (L:D) h and 0:24 (L:D) h than other photoperiods. As the scotophase increased, the preoviposition period decreased and the oviposition period increased. Female longevity exceeded that of males at all photoperiods, and both males and females at 0:24 (L:D) h had the shortest longevity. The number of eggs laid per female increased with increasing food consumption. Females at 8:16 (L:D) h had the greatest food consumption and laid the most eggs, while females at 24:0 (L:D) h had the lowest food consumption and laid few eggs. The food intake of adults increased gradually and decreased slowly after reaching a peak. Females began to lay eggs when their food consumption reached a maximum. These results indicate that a scotophase is necessary for the reproduction of H. oblita. A long scotophase promotes greater oviposition. The effect of photoperiod on reproduction is affected by food intake.

Genome-wide identification, characterization and expression analysis of the carotenoid cleavage oxygenase (CCO) gene family in Saccharum.

Carotenoid cleavage oxygenases (CCOs) play crucial roles in plant growth and development, as well as in the response to phytohormonal, biotic and abiotic stresses. However, comprehensive and systematic research on the CCO gene family has not yet been conducted in Saccharum. In this study, 47 SsCCO and 14 ShCCO genes were identified and characterized in Saccharum spontaneum and Saccharum spp. R570 cultivar, respectively. The SsCCOs consisted of 38 SsCCDs and 9 SsNCEDs, while ShCCOs contained 11 ShCCDs and 3 ShNCEDs. The SsCCO family could be divided into 7 groups, while ShCCO family into 5 groups. The genes/proteins contained similar compositions within the same group, and the evolutionary mechanisms differed between S. spontaneum and R570. Gene Ontology annotation implied that CCOs were involved in many physiological and biochemical processes. Additionally, 41 SsCCOs were regulated by 19 miRNA families, and 8 ShCCOs by 9 miRNA families. Cis-regulatory elements analysis suggested that CCO genes functioned in the process of growth and development or under the phytohormonal, biotic and abiotic stresses. qRT-PCR analysis indicated that nine CCO genes from different groups exhibited similar expression patterns under abscisic acid treatment, while more divergent profiles were observed in response to Sporisorium scitamineum and cold stresses. Herein, comparative genomics analysis of the CCO gene family between S. spontaneum and R570 was conducted to investigate its evolution and functions. This is the first report on the CCO gene family in S. spontaneum and R570, thus providing valuable information and facilitating further investigation into its function in the future.

A class III WRKY transcription factor in sugarcane was involved in biotic and abiotic stress responses.

WRKY transcription factors play significant roles in plant stress responses. In this study, a class III WRKY gene ScWRKY5, was successfully isolated from sugarcane variety ROC22. The ScWRKY5 was a nucleus protein with transcriptional activation activity. The ScWRKY5 gene was constitutively expressed in all the sugarcane tissues, with the highest expression level in the stem epidermis and the lowest in the root. After inoculation with Sporisorium scitamineum for 1 d, the expression level of ScWRKY5 was significantly increased in two smut-resistant varieties (YZ01-1413 and LC05-136), while it was decreased in three smut-susceptible varieties (ROC22, YZ03-103, and FN40). Besides, the expression level of ScWRKY5 was increased by the plant hormones salicylic acid (SA) and abscisic acid (ABA), as well as the abiotic factors polyethylene glycol (PEG) and sodium chloride (NaCl). Transient overexpression of the ScWRKY5 gene enhanced the resistance of Nicotiana benthamiana to the tobacco bacterial pathogen Ralstonia solanacearum, however the transiently overexpressed N. benthamiana was more sensitive to the tobacco fungal pathogen Fusarium solani var. coeruleum. These results provide a reference for further research on the resistance function of sugarcane WRKY genes.

New insights into the evolution and functional divergence of the CIPK gene family in Saccharum.

BACKGROUND: Calcineurin B-like protein (CBL)-interacting protein kinases (CIPKs) are the primary components of calcium sensors, and play crucial roles in plant developmental processes, hormone signaling transduction, and in the response to exogenous stresses. RESULTS: In this study, 48 CIPK genes (SsCIPKs) were identified from the genome of Saccharum spontaneum. Phylogenetic reconstruction suggested that the SsCIPK gene family may have undergone six gene duplication events from the last common ancestor (LCA) of SsCIPKs. Whole-genome duplications (WGDs) served as the driving force for the amplification of SsCIPKs. The Nonsynonymous to synonymous substitution ratio (Ka/Ks) analysis showed that the duplicated genes were possibly under strong purifying selection pressure. The divergence time of these duplicated genes had an average duplication time of approximately 35.66 Mya, suggesting that these duplication events occurred after the divergence of the monocots and eudicots (165 Mya). The evolution of gene structure analysis showed that the SsCIPK family genes may involve intron losses. Ten ScCIPK genes were amplified from sugarcane (Saccharum spp. hybrids). The results of real-time quantitative polymerase chain reaction (qRT-PCR) demonstrated that these ten ScCIPK genes had different expression patterns under abscisic acid (ABA), polyethylene glycol (PEG), and sodium chloride (NaCl) stresses. Prokaryotic expression implied that the recombinant proteins of ScCIPK3, - 15 and - 17 could only slightly enhance growth under salinity stress conditions, but the ScCIPK21 did not. Transient N. benthamiana plants overexpressing ScCIPKs demonstrated that the ScCIPK genes were involved in responding to external stressors through the ethylene synthesis pathway as well as to bacterial infections. CONCLUSIONS: In generally, a comprehensive genome-wide analysis of evolutionary relationship, gene structure, motif composition, and gene duplications of SsCIPK family genes were performed in S. spontaneum. The functional study of expression patterns in sugarcane and allogenic expressions in E. coli and N. benthamiana showed that ScCIPKs played various roles in response to different stresses. Thus, these results improve our understanding of the evolution of the CIPK gene family in sugarcane as well as provide a basis for in-depth functional studies of CIPK genes in sugarcane.

Evaluation of reference genes for quantitative real-time PCR normalization in the scarab beetle Holotrichia oblita.

Quantitative real-time polymerase chain reaction (qPT-PCR) is commonly used to analyze gene expression, however, the accuracy of the normalized results is affected by the expression stability of reference genes. Holotrichia oblita (Coleoptera: Scarabaeidae) causes serious damage to crops. Reliable reference genes in H. oblita are needed for qRT-PCR analysis. Therefore, we evaluated 13 reference genes under biotic and abiotic conditions. RefFinder provided a comprehensive stability ranking, and geNorm suggested the optimal number of reference genes for normalization. RPL13a and RPL18 were the most suitable reference genes for developmental stages, tissues, and temperature treatments; RPL13a and RPS3 were the most suitable for pesticide and photoperiod treatments; RPS18 and RPL18 were the most suitable for the two sexes. We validated the normalized results using odorant-binding protein genes as target genes in different tissues. Compared with the selected suitable reference genes, the expression of OBP1 in antennae, abdomen, and wings, and OBP2 in antennae and wings were overestimated due to the instability of ACTb. These results identified several reliable reference genes in H. oblita for normalization, and are valuable for future molecular studies.