Leaves are the main places for photosynthesis and organic synthesis of cotton.  Leaf shape has important effects on the photosynthetic efficiency and canopy formation, thereby affecting cotton yield.  Previous studies have shown that LMI1 is the main gene regulating leaf shape. In this study, the LMI1 gene (LATE MERISTEM IDENTITY1) was inserted into the 35S promoter expression vector, and cotton plants overexpressing LMI1(OE) were obtained through genetical transformation.  Statistical analysis of the biological traits of T₁ and T₂ populations showed that compared to wild type (WT), OE plants had significant larger leaves, thicker stems and significantly increased dry weight.  Furthermore, plant sections of the main vein and petiole showed that the number of cell in those tissues of OE plants increased significantly.  In addition, RNA-seq analysis revealed differential expression of genes related to gibberellin synthesis and NAC gene family (genes containing the NAC domain) in OE and WT plants, suggesting that LMI1 is involved in secondary wall formation and cell proliferation, and promotes stem thickening.  Moreover, GO (Gene Ontology) analysis enriched the terms of calcium ion binding, and KEGG (Kyoto Encyclopedia of Genes and Genomes) analysis enriched the terms of fatty acid degradation, phosphatidylinositol signal transduction system, and cAMP signal pathway.  These results suggested that LMI1 OE plants were responsive to gibberellin hormone signals, and altered messenger signal (cAMP, Ca²⁺) which amplified this function, to promote the stronger above ground vegetative growth.  This study found the LMI1 soared the nutrient growth in cotton, which is the basic for higher yield.

Maize (Zea mays L.) is a monoecious grass species with separate male and female inflorescences which form the tassel and ear, respectively.  The mature ear inflorescences usually bear hundreds of grains, thus directly influence maize grain production and yield.  Here, we isolated a recessive maize mutant, tasselseed2016 (ts2016), which exhibits pleiotropic inflorescence defects and a reduction in grain yield.  These defects include loss of determinacy and identity in meristems and floral organs, as well as a lack of the lower floret abortion in maize ear, and the smaller grain size.  Using map-based cloning and allelic test, we identified and confirmed a microRNA gene MIR172e as the target gene controlling these related traits.  Furthermore, our evidence uncovered a new potential miR172e/ETHYLENE RESPONSIVE ELEMENT BINDING197 (EREB197) regulatory module which controls the abortion of lower floret in maize ear. Transcriptome analysis revealed that the mutation of MIR172e represses multiple biological processes, particularly the flower development and hormone-related pathways in maize ear.  Additionally, we found the mutation in the DNA sequence of MIR172e affects in RNA transcription, resulting in elongation blockage at the mutant site.  Our results reveal the function and molecular mechanism of MIR172e in maize inflorescences and grain yield, and this study deepens our knowledge of maize inflorescence development.

Dry-hot wind stress causes losses in wheat productivity in major growing regions worldwide, especially for winter wheat in the Huang-Huai-Hai Plain of China, and both the occurrence and the severity of such events are likely to increase with global climate change.  To investigate the recovery of physiological function and yield formation using a non-commercial new chemical regulator (NCR) following dry-hot wind stress, we conducted a three-year field experiment (2018-2021), and sprayed tap water (control), monopotassium phosphate (CKP), NCR at both the jointing and flowering stage (CFS), and NCR only at the jointing stage (FSJ) or flowering stage (FSF).  The leaf physiology, biomass accumulation and translocation, grain-filling process, and yield components in winter wheat were investigated.  Among the single spraying treatments, the FSJ treatment was beneficial to accumulation of dry matter before anthesis, and to a larger increase of maximum grain-filling rate and mean grain-filling rate.  The FSF treatment performed better in maintaining a high relative chlorophyll content as SPAD value, and low rate of excised leaf water loss in flag leaves, promoting dry matter accumulation and contribution to grain after anthesis, prolonging the duration of grain filling, and inducing the period until maximum grain-filling rate to be reached earlier.  The CFS treatment was better than any other treatments in relieving effects of dry-hot wind.  The exogenous NCR treatments significantly increased grain yields by 12.45–18.20% in 2018–2019, 8.89–13.82% in 2019–2020, and 8.10–9.00% in 2020–2021.  The conventional measure of CKP treatment only increased grain yield by 6.69% in 2020–2021.  The CFS treatment had the greatest mitigating effect on loss of yield under dry-hot wind stress, following by the FSF and FSJ treatments, and the CKP treatment had a minimal effect.  In summary, the CFS treatment could be used as the main chemical control measure for wheat stress resistance and yield stability in areas with high incidence of dry-hot wind.  This can effectively regulate green retention and water status of leaves, promote dry matter accumulation and efficient translocation, improve grain-filling process, and ultimately reduce yield losses.

Seed aging tolerance during storage is generally an important trait for crop production, yet the role of small auxin-up RNA genes in conferring seed aging tolerance is largely unknown in rice.  In this study, one small auxin-up RNA gene, OsSAUR33, was found to be involved in the regulation of seed aging tolerance in rice.  The expression of OsSAUR33 was significantly induced in aged seeds compared with unaged seeds during the seed germination phase.  Accordingly, the disruption of OsSAUR33 significantly reduced seed vigor compared to the wild type (WT) in response to natural storage or artificial aging treatments.  The rice OsSAUR33 gene promotes the vigor of aged seeds by enhancing their reactive oxygen species (ROS) level during seed germination, and the accumulation of ROS was significantly delayed in the aged seeds of Ossaur33 mutants in comparison with WT during seed germination.  Hydrogen peroxide (H₂O₂) treatments promoted the vigor of aged seeds in various rice varieties.  Our results provide timely theoretical and technical insights for the trait improvement of seed aging tolerance in rice.

Cropland nitrate leaching is the major nitrogen (N) loss pathway, and it contributes significantly to water pollution. However, cropland nitrate leaching estimates show great uncertainty due to variations in input datasets and estimation methods. Here, we presented a re-evaluation of Chinese cropland nitrate leaching, and identified and quantified the sources of uncertainty by integrating three cropland area datasets, three N input datasets, and three estimation methods. The results revealed that nitrate leaching from Chinese cropland averaged 6.7±0.6 Tg N yr⁻¹ in 2010, ranging from 2.9 to 15.8 Tg N yr⁻¹ across 27 different estimates. The primary contributor to the uncertainty was the estimation method, accounting for 45.1%, followed by the interaction of N input dataset and estimation method at 24.4%. The results of this study emphasize the need for adopting a robust estimation method and improving the compatibility between the estimation method and N input dataset to effectively reduce uncertainty. This analysis provides valuable insights for accurately estimating cropland nitrate leaching and contributes to ongoing efforts that address water pollution concerns.

Chinese cabbage is an important leafy vegetable crop with high water demand and susceptibility to drought stress.  To explore the molecular mechanisms underlying the response to drought, we performed a transcriptome analysis of drought-tolerant and -sensitive Chinese cabbage genotypes under drought stress, and uncovered core drought-responsive genes and key signaling pathways.  A co-expression network was constructed by a weighted gene co-expression network analysis (WGCNA) and candidate hub genes involved in drought tolerance were identified.  Furthermore, abscisic acid (ABA) biosynthesis and signaling pathways and their drought responses in Chinese cabbage leaves were systemically explored.  We also found that drought treatment increased the antioxidant enzyme activities and glucosinolate contents significantly.  These results substantially enhance our understanding of the molecular mechanisms underlying drought responses in Chinese cabbage.

Rapeseed (Brassica napus L.) is the second most widely grown premium oilseed crop globally, mainly for its vegetable oil and protein meal.  One of the main goals of breeders is producing high-yield rapeseed cultivars with sustainable production to meet the requirements of the fast-growing population.  Besides the pod number, seeds per silique (SS), and thousand-seed weight (TSW), the ovule number (ON) is a decisive yield determining factor of individual plants and the final seed yield.  In recent years, tremendous efforts have been made to dissect the genetic and molecular basis of these complex traits, but relatively few genes or loci controlling these traits have been reported thus far.  This review highlights the updated information on the hormonal and molecular basis of ON and development in model plants (Arabidopsis thaliana).  It also presents what is known about the hormonal, molecular, and genetic mechanism of ovule development and number, and bridges our understanding between the model plant species (A. thaliana) and cultivated species (B. napus).  This report will open new pathways for primary and applied research in plant biology and benefit rapeseed breeding programs.  This synopsis will stimulate research interest to further understand ovule number determination, its role in yield improvement, and its possible utilization in breeding programs. 

Plant chlorophyll biosynthesis and chloroplast development are two complex processes that are regulated by exogenous and endogenous factors.  In this study, we identified OsDXR, a gene encoding a reductoisomerase that positively regulates chlorophyll biosynthesis and chloroplast development in rice.  OsDXR knock-out lines displayed the albino phenotype and could not complete the whole life cycle process.  OsDXR was highly expressed in rice leaves, and subcellular localization indicated that OsDXR is a chloroplast protein.  Many genes involved in chlorophyll biosynthesis and chloroplast development were differentially expressed in the OsDXR knock-out lines compared to the wild type.  Moreover, we found that the RNA editing efficiencies of ndhA-1019 and rpl2-1 were significantly reduced in the OsDXR knock-out lines.  Furthermore, OsDXR interacted with the RNA editing factor OsMORF1 in a yeast two-hybrid screen and bimolecular fluorescence complementation assay.  Finally, disruption of the plastidial 2-C-methyl-derythritol-4-phosphate pathway resulted in defects in chloroplast development and the RNA editing of chloroplast genes.

The physiological and metabolic differences in maize under different nitrogen (N) levels are the basis of reasonable N management, which is vital in improving fertilizer utilization and reducing environmental pollution.  In this paper, on the premise of defining the N fertilizer efficiency and yield under different long-term N fertilization treatments, the corresponding differential metabolites and their metabolic pathways were analyzed by untargeted metabolomics in maize.  N stress, including deficiency and excess, affects the balance of carbon (C) metabolism and N metabolism by regulating C metabolites (sugar alcohols and tricarboxylic acid (TCA) cycle intermediates) and N metabolites (various amino acids and their derivatives).  L-alanine, L-phenylalanine, L-histidine, and L-glutamine decreased under N deficiency, and L-valine, proline, and L-histidine increased under N excess.  In addition to sugar alcohols and the above amino acids in C and N metabolism, differential secondary metabolites, flavonoids (e.g., kaempferol, luteolin, rutin, and diosmetin), and hormones (e.g., indoleacetic acid, trans-zeatin, and jasmonic acid) were initially considered as indicators for N stress diagnosis under this experimental conditions.  This study also indicated that the leaf metabolic levels of N2 (120 kg ha^–1 N) and N3 (180 kg ha^–1 N) were similar, consistent with the differences in their physiological indexes and yields over 12 years.  This study verified the feasibility of reducing N fertilization from 180 kg ha^–1 (locally recommended) to 120 kg ha^–1 at the metabolic level, which provided a mechanistic basis for reducing N fertilization without reducing yield, further improving the N utilization rate and protecting the ecological environment.

Natural rubber (NR) is an irreplaceable biopolymer of economic and strategic importance owing to its unique physical and chemical properties.  The Pará rubber tree (Hevea brasiliensis (Willd. ex A. Juss.) Müll. Arg.) is currently the exclusive commercial source of NR, and it is primarily grown in plantations restricted to the tropical and subtropical areas of Southeast Asia.  However, current Pará rubber production barely meets the sharply increasing global industrial demand for rubber.  Petroleum-based synthetic rubber (SR) has been used to supplement the shortage of NR but its industrial performance is not comparable to that of NR.  Thus, there is an urgent need to develop new productive rubber crops with broader environmental adaptability.  This review summarizes the current research progress on alternative rubber-producing plants, including horticultural plants (Taraxacum kok-saghyz Rodin and Lactuca L. species), woody plants (Parthenium argentatum A. Gray and Eucommia ulmoides Oliv.), and other plant species with potential for NR production.  With an emphasis on the molecular basis of NR biosynthesis revealed by a multi-omics approach, we highlight new integrative strategies and biotechnologies for exploring the mechanism of NR biosynthesis with a broader scope, which may accelerate the breeding and improvement of new rubber crops. 

Antimicrobial resistance has become a global problem that poses great threats to human health.  Antimicrobials are widely used in broiler chicken production and consequently affect their gut microbiota and resistome.  To better understand how continuous antimicrobial use in farm animals alters their microbial ecology, we used a metagenomic approach to investigate the effects of pulsed antimicrobial administration on the bacterial community, antibiotic resistance genes (ARGs) and ARG bacterial hosts in the feces of broiler chickens.  Chickens received three 5-day courses of individual or combined antimicrobials, including amoxicillin, chlortetracycline and florfenicol.  The florfenicol administration significantly increased the abundance of mcr-1 gene accompanied by floR gene, while amoxicillin significantly increased the abundance of genes encoding the AcrAB-tolC multidrug efflux pump (marA, soxS, sdiA, rob, evgS and phoP).  These three antimicrobials all led to an increase in Proteobacteria.  The increase in ARG host, Escherichia, was mainly attributed to the β-lactam, chloramphenicol and tetracycline resistance genes harbored by Escherichia under the pulsed antimicrobial treatments.  These results indicated that pulsed antimicrobial administration with amoxicillin, chlortetracycline, florfenicol or their combinations significantly increased the abundance of Proteobacteria and enhanced the abundance of particular ARGs.  The ARG types were occupied by the multidrug resistance genes and had significant correlations with the total ARGs in the antimicrobial-treated groups.  The results of this study provide comprehensive insight into pulsed antimicrobial-mediated alteration of chicken fecal microbiota and resistome.

Nipah virus (NiV) is a member of the genus Henipavirus of the family Paramyxoviridae, and is an emerging zoonotic virus (Singh et al. 2019). NiV was first observed in 1998 and this outbreak triggered an uncontrollable epidemic lasting until 1999, during which more than a million pigs were destroyed (Ang et al. 2018). Since then, several South and Southeast Asian countries have experienced relatively small sporadic outbreaks almost every year, with mortality rates of up to 40-75 % in humans infected with NiV (Raj Kumar et al. 2019). Therefore, NiV not only does terrible damage to the farming industry but is also a serious threat to human public health and safety. Fruit bats are recognized as natural reservoir hosts for NiV, and other species of bats, as well as pigs, horses, dogs, and humans can also be infected (Soman Pillai et al. 2020). Bats act as asymptomatic carriers of the virus, but they excrete the virus through their saliva, urine, semen, and feces (Soman Pillai et al. 2020). NiV infection in humans and animals primarily occurs through the consumption of date palm juice and fruits that have been contaminated by fruit bats (Bruno et al. 2022). Human-to-human transmission is also possible (Garbuglia et al. 2023). NiV infection in humans and pigs can result in fatal encephalitis and severe respiratory disease (Ma et al. 2019, Ming-Yen et al. 2020). Because of its high pathogenicity and mortality, as well as the lack of effective treatments or vaccines for humans or animals (Gómez Román et al. 2022), NiV is included in the list of epidemiological threats requiring urgent R&D action in the World Health Organization (WHO) R&D Blueprint as one of the pathogens. WHO has therefore prepared a technical brief aimed at guiding countries in their preparedness planning for a NiV event, particularly in countries that have not yet reported a NiV event. The technical brief suggests that early diagnosis of NiV can enhance the survival chances of an infected individual and can also prevent transmission to others.

Currently, the detection methods for NiV are mainly based on traditional serological methods and molecular diagnostic methods. Of the serological detection methods, ELISA, phage plaque assay, and immunofluorescence staining do not require special instruments or equipment, but the NiV-infected samples need to be processed in a Biosafety-Level-4 (BSL-4) laboratory, which is risky for operators and is not available in all regions (Diwakar D et al. 2015, Fischer et al. 2018, Gary et al. 2001). Real-time reverse transcriptase polymerase chain reaction (RT-PCR) tests are the preferred method for viral detection due to their high sensitivity and ability to detect the virus in patients at the earliest stages of infection. However, Real-time RT-PCR requires sophisticated instruments and equipment, as well as specialized operator personnel, and cannot fully satisfy the need for early diagnosis of NiV infection in situ or in poor areas lacking these conditions (Wu et al. 2022). Therefore, the development of a rapid, specific, and sensitive assay that does not require complex instrumentation is critical for the early detection of NiV.

Nucleic acid isothermal amplification technology, characterized by its rapidity, sensitivity, and specificity, is capable of meeting the demands for speed and simplicity detection, and has significant practical application value (Zhao et al. 2015). Of these methods, recombinase-aided amplification (RAA) technology, with its simplicity and ease of use, is ideal for early diagnosis in resource-limited settings (James et al. 2018). Recombinant enzyme-based isothermal amplification tests have been used to develop three rapid assays for NiV that can detect 1,000 copies μL^-1 for synthetic NiV RNA in less than 30 min (Pollak et al. 2023). However, these assays require the transfer of the RPA amplification products, exposing the nucleic acid-rich samples to the environment, and leading to potential aerosol contamination. In the present study, a fully enclosed device was employed, combining reverse transcription recombinase-aided amplification technology with lateral flow immunochromatography (RT-RAA-VF), to establish a rapid and sensitive assay for the detection of NiV. This assay avoids false positives caused by aerosols, does not require sophisticated instruments and equipment, does not require specialized personnel to operate, and holds the potential to fulfill the requirements for early diagnosis during the initial stages of a NiV infection outbreak even in poor and remote areas.

To detect all known NiV strains, 55 P gene sequences of different NiV strains, including NiV-B (transmitted in Bangladesh and India), NiV-M (transmitted in Malaysia), and NiV-T (transmitted in Thailand) were retrieved from GenBank (https: //www.ncbi.nlm.nih.gov/) and compared using MAFFT version 7. The highly conserved region of the P gene was selected as the target and was used to design the specific RAA primers and probe by the Prime Primer 5.0 software. The probe contained 55 nucleotides with the tetrahydrofuran (THF) residue replacing the adenine at position 37. The 5' end of the probe was labeled with the fluorescent marker FAM, while the 3' end was labeled with a blocking group comprising a C3 spacer. The 5' end of the reverse primer was labeled with biotin (Table 1). The primers and probe were synthesized by Sangon Biotech Co., Ltd. (Shanghai, China). The RNA transcript of NiV P (Genebank: NC_002728) and the positive plasmid pcDNA3.1-NiV-P were also synthesized by Sangon Biotech. In the presence of NiV RNA, the reverse transcriptase is activated to reverse transcribe the NiV RNA to cDNA, and the RAA amplification reaction is followed activated by two priming oligonucleotides to produce targets for probe annealing hybridization. The THF site of the probe can then be cleaved by the nfo enzyme to create a new 3'-hydroxyl group, which is used as a start site for polymerase extension, thus converting the probe into a primer that, together with a reverse primer with biotin, produces double-stranded amplification products labeled with two antigens (FAM and biotin). Subsequently, the reaction tube, which contains the amplified products, is positioned within a sealed and disposable nucleic acid visualization paper device. The amplified DNA products are capable of adhering to streptavidin-coated gold nanoparticles, thereby forming a conjugate that is subsequently immobilized by the anti-FAM antibody present on the test line (T line), which manifests as a red stripe. Meanwhile, any streptavidin-labeled gold nanoparticles that are not associated with the amplified DNA are trapped by the anti-streptavidin antibody at the control line (C line) (Fig. 1-A).

To find the optimal RT-RAA reaction time, 10-fold gradient dilutions of the plasmid pcDNA3.1-NiV-P (5×10¹⁰ copies μL^-1) were used as targets for RT-RAA-VF assay. The reaction was performed at three distinct temperatures—42 °C, 39 °C, and 37 °C—for a consistent period of 20 min. The outcomes indicated that at 42 °C, the emergence of discernible bands on both the C and T lines, indicating of a positive result, was observed at plasmid concentrations of 5 copies μL^-1 or greater. At 39 °C and 37 °C, the positive results were only observed at plasmid concentrations of 500 copies μL^-1 or more. These results suggested that 42 °C was the optimal reaction temperature for the NiV RT-RAA assay (Fig. 1-B). To determine the most efficient amplification time, the assay was executed for time intervals of 15, 20, 25, and 30 min. Analysis of the data provided in Appendix A confirmed that the optimal amplification duration for the assay was 20 min.

Next, we assessed the sensitivity of the assay using 10-fold gradient dilutions of NiV P RNA transcripts, reacting at 42 °C for 20 min. The results showed that this assay could detect as few as 5 copies μL^-1 of RNA transcripts (Fig. 1-C). We further compared the sensitivity of this assay with that of the real-time RT-PCR method currently recommended by the World Organisation for Animal Health (WOAH), using 10-fold serial dilutions of RNA extracted from NiV-infected Vero E6 cells as templates.  The results showed that the two methods shared an identical detection limit, and both were capable of detecting NiV RNA from infected cells diluted 10,000 times (Appendix B). To evaluate the specificity of this assay, the nucleic acids of pathogens that can cause neurological symptoms similar to NiV were also tested, including Hendra virus (HeV), herpes simplex virus type 1 (HSV-1), rabies virus (RABV) and Streptococcus suis. The results showed that our established assay only recognized the nucleic acids of NiV, with no cross-reactivity observed with other pathogens (Fig. 1-D).

It has been reported that NiV can be transmitted by droplets, or by contact with throat or nasal secretions, from the respiratory tract of patients or sick pigs. To validate the clinical applicability of this assay, we mixed NiV RNA with RNA extracted from human saliva samples to simulate NiV-infected clinical samples. The simulated NiV clinical samples were evaluated using the RT-RAA-VF assay as well as real-time RT-PCR. Of all 47 samples tested, 25 samples were prepared by 3-, 4-, and 5-fold gradient dilutions of NiV RNA using healthy human RNA as the diluent, and 22 samples were healthy human RNA samples without NiV RNA. The RT-RAA-VF assay was able to distinguish between the 25 positive and 22 negative samples, and the real-time RT-PCR assay also detected 25 positive and 22 negative samples, with Ct values ranging from 25 to 40 for positive samples (Fig. 1-E to G). We also simulated the clinical NiV infected swine samples. Among all 28 samples, 15 samples were obtained by 3-, 4- and 5-fold gradient dilutions of NiV RNA with healthy swine RNA as the diluent, and 13 samples were healthy swine RNA samples without NiV RNA. The results demonstrated that the RT-RAA-VF assay was in accordance with the real-time RT-PCR and was capable of effectively differentiating between 15 positive samples and 13 negative samples (Fig 1-H to J). The assay is therefore expected to be an alternative assay for the clinical detection of NiV, being faster, more specific, and more sensitive than real-time RT-PCR and without the need for complex instrumentation.

As human societies evolve, the combination of urbanization and climate change has resulted in the destruction of bat habitats, leading to increased contact between bats and humans. This has further elevated the risk of NiV infection for both humans and animals (Jonathan A et al. 2004, Kessler et al. 2018). NiV outbreaks have been reported in four countries - Bangladesh, India, Malaysia, and Singapore. However, fruit bats (the natural reservoir host of NiV) have been found in several Southeast Asian countries, suggesting the potential for NiV outbreaks in previously unaffected areas (Mangesh et al. 2022). Given the incubation period associated with NiV, the potential for cross-regional spread due to the movement of infected individuals cannot be dismissed. Consequently, it is imperative to focus on the rapid detection of NiV infections to enhance the survival prospects for those affected and to mitigate the risk of further transmission of the virus to others. In the present study, we developed an RT-RAA-VF assay for the detection of NiV using a sealed disposable nucleic acid visualization test paper device which effectively eliminates false positives caused by aerosol contamination. The assay demonstrated a sensitivity of 5 copies μL^-1 for detecting NiV RNA at 42 °C for 20 minutes. The assay exhibits high specificity, with no cross-reactivity observed against other paramyxoviruses or neurological pathogens. Moreover, when evaluated on simulated clinical samples, it demonstrated 100% concordance with the results obtained from the real-time RT-PCR method. With its rapidity, specificity, and sensitivity features, this assay holds promise as an effective tool for early diagnosis of NiV infection.

Crop improvement is crucial for addressing the global challenges of food security and sustainable agriculture.  Recent advancements in high-throughput phenotyping technologies and artificial intelligence (AI) have revolutionized the field, enabling rapid and accurate assessment of crop traits on a large scale.  The integration of AI and machine learning algorithms with high-throughput phenotyping data has unlocked new opportunities for crop improvement.  AI algorithms can analyze and interpret large datasets, extracting meaningful patterns and correlations between phenotypic traits and genetic factors.  These technologies have the potential to revolutionize plant breeding programs by providing breeders with efficient and accurate tools for trait selection, reducing the time and cost required for variety development.  However, further research and collaborations are needed to overcome the challenges and fully unlock the power of high-throughput phenotyping and AI in crop improvement.  By leveraging AI algorithms, researchers can efficiently analyze phenotypic data, uncover complex patterns, and establish predictive models that enable precise trait selection and crop breeding.  The aim of this review is to explore the transformative potential of integrating high-throughput phenotyping and AI in crop improvement.  The review will encompass an in-depth analysis of recent advancements and applications, highlighting the numerous benefits and challenges associated with high-throughput phenotyping and intelligence.

Elevating soil water content (SWC) through irrigation was one of the simple mitigation measures to improve crop resilience to heat stress.  The response of leaf function such as photosynthetic capacity based on chlorophyll fluorescence during the mitigation has received limited attention, especially in field condition. The 2-yr field experiment with three treatments (control treatment (CK); high temperature treatment (H); high temperature together with elevating SWC treatment (HW)) was carried out during grain filling with two maize hybrids at a typical station in North China Plain.  Averagely, the net photosynthetic rate (P_n) was improved by 20% in HW treatment and a 1-3°C decrease in canopy temperature compared with H treatment in two years. Furthermore, the higher SWC in HW treatment significantly improved actual photosynthetic rate (Phi2), linear electron flow (LEF), variable fluorescence (F_v) and maximal potential quantum efficiency (F_v/F_m) for both hybrids.  Meanwhile, different responses in chlorophyll fluorescence between hybrids were also observed.  The higher SWC in HW treatment significantly improved thylakoid proton conductivity (gH⁺) and maximal fluorescence (F_m) for the hybrid ZD958.  For the hybrid XY335, the proton conductivity of chloroplast ATP synthase (vH⁺) and minimal fluorescence (F₀) was increased by the SWC.  The SEM model further showed that SWC had significantly positive relationships with P_n, LEF, and F_v/F_m.  The elevating SWC alleviated heat stress with the delayed leaf senescence to prolong the effective period of photosynthesis and enhanced leaf photosynthetic capacity by improving Phi2, LEF, F_v and F_v/F_m.  This research demonstrates that elevating SWC through enhancing leaf photosynthesis during grain filling would be an import mitigation measure to adapt to warming climate in maize production.

Maize (Zea mays L.) is an economically vital grain crop that is cultivated worldwide. In 2011, a maize foliar disease was detected in Lingtai and Lintao counties in Gansu Province, China. The characteristic signs and symptoms of this disease include irregular chlorotic lesions on the tips and edges of infected leaves and black punctate fruiting bodies in dead leaf tissues. Given favourable environmental conditions, this disease spread to areas surrounding Gansu. In this study, infected leaves were collected from Gansu and Ningxia Hui Autonomous Region between 2018 and 2020 to identify the disease-causing pathogen. Based on morphological features; pathogenicity tests; and multi-locus phylogenetic analysis involving internal transcribed spacer (ITS), 18S small subunit rDNA (SSU), 28S large subunit rDNA (LSU), translation elongation factor 1-alpha (TEF), and β-tubulin (TUB) sequences, Eutiarosporella dactylidis was identified as the causative pathogen of this newly discovered leaf blight. Furthermore, an in vitro bioassay was conducted on representative strains using six fungicides, and both fludioxonil and carbendazim were found to significantly inhibit the mycelial growth of E. dactylidis. The results of this study provide a reference for the detection and management of Eutiarosporella leaf blight.

Streptococcus agalactiae is one of the most common pathogens that cause bovine mastitis worldwide. Identifying pathogen prevalence and virulence factors is critical for developing prevention and control approaches. Herein, 1161 milk samples from various dairy farms in China (n=558) and Pakistan (n=603) were collected between 2019-2021 and were subjected to S. agalactiae isolation. Prevalence, serotyping, virulence genes, and antibiotic-resistant genes of S. agalactiae were evaluated by PCR assay. All isolates were characterized for haemolysis, biofilm production, cytotoxicity, adhesion, and invasion on bovine mammary epithelial cells. The prevalence of S. agalactiae-induced mastitis in cattle was found to be considerably higher in Pakistan than in China. Jiangsu and Sindh provinces had the highest area-wise prevalence in China and Pakistan, respectively. Serotypes Ia and II were prevalent in both countries, whereas serotype III was found only in Pakistan. Moreover, all isolates tested positive for PI-2b gene but negative for PI-1 and PI-2a genes. All isolates harboured cfb, cylE, hylB, and fbsB virulent genes, whereas many of them lacked bibA, rib and bca. However, the absence of bac and scp genes in Chinese isolates and cspA in Pakistani isolates was noted, while spb1 and lmb were not detected in isolates of both countries. Pakistani isolates, particularly serotype Ia-positive, had a considerably higher ability to produce biofilm, haemolysis, cytotoxicity, adhesion, and invasion than Chinese isolates. Most of the isolates were phenotypically resistant to tetracycline, erythromycin, and clindamycin and genotypic resistance was confirmed by the presence of ermA, ermB, tetM and tetO genes. Our study highlights the antimicrobial resistance profile and virulence-related factors contributing to the epidemiological spread of mastitis-causing S. agalactiae in China and Pakistan. The findings may facilitate future studies designed to develop improved treatment and control strategies against this pathogen. 

Plant height is a key plant architectural trait that affects the seed yield, harvest index and lodging resistance in Brassica napus L., although the genetic mechanisms affecting plant height remain unclear.  Here, a semi-dwarf mutant, df34, was obtained by ethyl methanesulphonate-induced mutagenesis.  Genetic analysis showed that the semi-dwarf phenotype is controlled by one semi-dominant gene, which was located on chromosome C03 using a bulked segregant analysis coupled with whole-genome sequencing, and this gene was named BnaSD.C3.  Then BnaSD.C3 was fine-mapped to a 297.35-kb segment of the “Darmor-bzh” genome, but there was no potential candidate gene for the semi-dwarf trait underlying this interval.  Furthermore, the interval was aligned to the Zhongshuang 11 reference genome.  Finally, combining structural variation analysis, transcriptome sequencing, phytohormone analyses and gene annotation information, BnaC03G0466900ZS and BnaC03G0478900ZS were determined to be the most likely candidate genes affecting the plant height of df34.  This study provides a novel major locus for breeding and new insights into the genetic architecture of plant height in B. napus

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a devastating pest of citrus fruits.  After successful mating, adult females insert their eggs into the ripened fruit, resulting in moldy and rotten fruit and causing great economic losses for the citrus industry.  In the field, flies initiate copulatory behaviors as twilight approaches, and decreasing light intensity in this period is the normal stimulus for copulation.  In this study, ten light intensities ranging from 0–30 000 lux were set to identify the typical intensity that strongly regulates the copulation behavior of B. dorsalis.  Three light intensities found to regulate the copulation behavior were then selected to verify their effects on adult male wing fanning and female chemotaxis towards 2,3,5-trimethylpyrazine (TMP).  At last, strong light and complete darkness were artificially combined in the lab to verify whether they could prevent copulation to inform behavioral manipulation of oriental flies in the future.  The results indicated that adult flies generally initiated copulatory behaviors at low light intensity (<1 000 lux).  
Stronger light significantly prevented copulation in proportion to intensity, with nearly no copulation events initiated when light intensity was above 20 000 lux.  Both male wing fanning and female chemotaxis towards TMP were attenuated as light intensity became stronger.  However, at 10 000 lux, males still fanned their wings to a certain extent while TMP completely lost its attractiveness to females.  In the darkness, adults did not initiate any sexual behaviors, e.g., copulation, wing fanning, or chemotaxis to TMP.  One hour of strong light (10 000 lux) combined with continuous darkness completely prevented mating.  These results show that light condition is an essential factor for copulatory behaviors in the oriental fruit fly.  Researchers could thus manipulate light conditions artificially or disrupt the molecular target in flies’ light transduction pathway to develop environmentally-friendly techniques to control this pest.

Banana is a significant crop, and three banana leaf diseases, including Sigatoka, Cordana and Pestalotiopsis, have the potential to have a serious impact on banana production. Existing studies are insufficient to provide a reliable method for accurately identifying banana leaf diseases. Therefore, this paper proposes a novel method to identify banana leaf diseases. First, a new algorithm called K-scale VisuShrink algorithm (KVA) is proposed to denoise banana leaf images. The proposed algorithm introduces a new decomposition scale k based on the semi-soft and middle course thresholds, the ideal threshold solution is obtained and substituted with the newly established threshold function to obtain a less noisy banana leaf image. Then, this paper proposes a novel network for image identification called Ghost ResNeSt-Attention RReLU-Swish Net (GR-ARNet) based on Resnet50. In this, the Ghost Module is implemented to improve the network's effectiveness in extracting deep feature information on banana leaf diseases and the identification speed; the ResNeSt Module adjusts the weight of each channel, increasing the ability of banana disease feature extraction and effectively reducing the error rate of similar disease identification; the model's computational speed is increased using the hybrid activation function of RReLU and Swish. Our model achieves an average accuracy of 96.98% and a precision of 89.31% applied to 13021 images, demonstrating that the proposed method can effectively identify banana leaf diseases.

非洲猪瘟在我国爆发，给我国养殖业带来巨大的经济损失，目前无可用的商业化疫苗和药物，其相关研究限定于在高等级生物安全实验室中完成，生物安全防护中消毒剂的合理使用尤为重要。MICRO-CHEM PLUS（MCP）作为一种复合型季铵盐类消毒剂在高等级生物安全实验室广泛使用，而针对非洲猪瘟病毒（African swine fever Virus，ASFV）的灭活效果未见相关报道。本文我们探究不同病毒载量、不同消毒剂浓度、不同作用时间及不同作用温度对于灭活非洲猪瘟病毒效果的影响。研究结果表明高病毒载量需要更高浓度的MCP才能将ASFV完全灭活，较低浓度的MCP需要延长作用时间才能达到完全灭活ASFV的效果，不同的作用温度对于MCP灭活ASFV的效果无影响。应用干雾消毒机将5%MCP进行房间喷雾消毒，当浓度达到0.06L/m3，用ASFV、大肠杆菌和金色葡萄球菌作为生物指示剂，可以达到终末消毒效果，但对于枯草芽孢杆菌作为生物指示剂还有部分活菌残留；当浓度为0.03L/m3时，对于大肠杆菌或金黄色葡萄球菌作为生物指示剂，也可以达到了终末消毒效果。该研究为在特定环境中合理使用 MCP 提供了科学依据，可以用于操作ASFV的高等级生物安全实验室的消毒以及猪场非洲猪瘟感染的预防。

Passion fruit (Passiflora edulis Sims) is a vine of the Passiflora genus in the Passifloraceae family.  The extracted components include flavonoids and terpenoids, which have good anti-anxiety and anti-inflammatory effects in humans.  In this study, we analyzed the transcriptomes of four tissues of the ‘Zixiang’ cultivar using RNA-Seq, which provided a dataset for functional gene mining.  The de novo assembly of these reads generated 96 883 unigenes, among which 61 022 unigenes were annotated (62.99% yield).  In addition to its edible value, another important application of passion fruit is its medicinal value.  The flavonoids and terpenoids are mainly derivatives of luteolin, apigenin, cycloartane triterpenoid saponins and other active substances in leaf extracts.  A series of candidate unigenes in the transcriptome data that are potentially involved in the flavonoid and terpenoid synthesis pathways were screened using homology-based BLAST and phylogenetic analysis.  The results showed that the biosynthesis of triterpenoids in passion fruit comes from the branches of the mevalonate (MVA) and 2-C-methyl-D-erythritol 4-phosphate/1-deoxy-D-xylulose 5-phosphate (MEP/DOXP) pathways, which is different from the MVA pathway that is used in other fruit trees.  Most of the candidate genes were found to be highly expressed in the leaves and/or flowers.  Quantitative real-time PCR (qRT-PCR) verification was carried out and confirmed the reliability of the RNA-Seq data.  Further amplification and functional analysis of these putative unigenes will provide additional insight into the biosynthesis of flavonoids and terpenoids in passion fruit.

Malus prunifolia Borkh. ‘Fupingqiuzi’ has significant ecological and economic value and plays a key role in germplasm development and resistance research.  However, its long juvenile phase and high heterozygosity are barriers to the identification of ‘Fupingqiuzi’ progeny with excellent traits.  In-vitro regeneration techniques and Agrobacterium-mediated genetic transformation systems can efficiently produce complete plants and thus enable studies of gene function.  However, optimal regeneration and genetic transformation systems for ‘Fupingqiuzi’ have not yet been developed.  Here, we evaluated the factors that affect the in-vitro regeneration and transformation of ‘Fupingqiuzi’.  The best results were obtained when transverse leaf sections were used as explants, and they were grown in dark culture for three weeks with their adaxial sides contacting the culture medium (MS basal salts, 30 g L⁻¹ sucrose, 8 g L⁻¹ agar, 5 mg L⁻¹  6-benzylaminopurine (6-BA), 2 mg L⁻¹ thidiazuron (TDZ), and 1 mg L⁻¹ 1-naphthlcetic acid (NAA), pH 5.8).  A genetic transformation system based on this regeneration system was optimized: after inoculation with A. tumefaciens solution for 8 min, 4 days of co-culture, and 3 days of delayed culture, the cultures were screened with cefotaxime (150 mg L⁻¹) and kanamycin (15 mg L⁻¹).  We thus established an efficient regeneration and genetic transformation system for ‘Fupingqiuzi’, enabling the rapid production of transgenic material.  These findings make a significant contribution to apple biology research

Ear-related traits are often selection targets for maize improvement.  This study used an immortalized F₂ (IF₂) population to elucidate the genetic basis of ear-related traits.  Twelve ear-related traits (namely, row number (RN), kernel number per row (KNPR), ear length (EL), ear diameter (ED), ten-kernel thickness (TKT), ear weight (EW), cob diameter (CD), kernel length (KL), kernel width (KW), grain weight per ear (GW), 100-kernel weight (HKW), and grain yield per plot (GY)), were collected from the IF₂ population.  The ear-related traits were comprised of 265 crosses derived from 516 individuals of the recombinant inbred lines (RILs) under two separated environments in 2017 and 2018, respectively.  Quantitative trait loci (QTLs) analyses identified 165 ear traits related QTLs, which explained phenotypic variation ranging from 0.1 to 12.66%.  Among the 165 QTLs, 19 underlying nine ear-related traits (CD, ED, GY, RN, TKT, HKW, KL, GW, and KNPR) were identified across multiple environments and recognized as reliable QTLs.  Furthermore, 44.85% of the total QTLs showed an overdominance effect, and 12.72% showed a dominance effect. Additionally, we found 35 genomic regions exhibiting pleiotropic effects across the whole maize genome, and 17 heterotic loci (HLs) for RN, EL, ED and EW were identified.  The results provide insights into genetic components of ear-related traits and enhance the understanding of the genetic basis of heterosis in maize. 

Wheat relative species are important for agriculture production, functional genomics study and wheat improvement as useful genetic resources.  In this study, a regeneration related wheat gene TaWOX5 was applied to establish the Agrobacterium-mediated transformation systems of Triticum monococcum, hexaploid triticale, and rye (Secale cereale L.) using their immature embryos.  Transgenic plants were efficiently generated.  During the transformation process, the Agrobacterium infection efficiency was assessed by histochemical staining for β-glucuronidase (GUS).  Finally, the transgenic nature of regenerated plants was verified by polymerase chain reaction (PCR)-based genotyping for presence of the GUS and bialaphos resistance (bar) genes, histochemical staining for GUS protein, and QuickStix strip assay for bar protein. The transformation efficiency of T. monococcum genotype PI428182 was 94.4%; the efficiencies of four hexaploid triticale genotypes Lin456, ZS3297, ZS1257, and ZS3224 were 52.1, 41.2, 19.4, and 16.0%, respectively; the transformation efficiency of rye cultivar Lanzhou Heimai was 7.8%.  Fluorescence in situ hybridization (FISH) and genomic in situ hybridization (GISH) analysis indicated that the GUS transgenes were integrated into the distal or near centromere (proximal) region of the chromosomes in transgenic T. monococcum and hexaploid triticale plants; in the transgenic hexaploid triticale plants, the foreign DNA fragment was randomly integrated into the AABB genome and RR genome.  Furthermore, the transgene was proved to be almost stably inherited in the next generation by Mendel’s law.  The findings in this study will promote genetic improvement for grain or forage production of the three plant species and for functional genomics study of cereal species including wheat.