A mutation in the promoter of the yellow stripe-like transporter gene in cucumber results in a yellow cotyledon phenotype

Leaf color mutants in higher plants are considered to be ideal materials for studying the chlorophyll biosynthesis,photosynthesis mechanism and chloroplast development.Herein,we identified a spontaneous mutant,yc412,in cultivated cucumber that exhibited yellow cotyledons.The yellow-lethal mutant was diagnosed with an abnormal chloroplast ultrastructure,and reduced photosynthetic capacity and pigment content.Through bulked segregant analysis-based whole-genome sequencing and fine genetic mapping,we narrowed the yellow cotyledons (yc) locus to a 96.8 kb interval on chromosome 3.By resequencing and molecular cloning,we showed that Csyc is a potential candidate gene,which encodes a yellow stripe-like (YSL) transporter.The T to C mutation in the promoter region of Csyc caused the yellow cotyledon phenotype in yc412.Compared to YZU027A (WT),the expression of Csyc was significantly downregulated in the cotyledons of yc412.Silencing of Csyc in cucumber via virus-induced gene silencing resulted in chlorotic leaves,mainly by suppressing the chlorophyll content.Furthermore,a comparative transcriptome analysis revealed that chloroplast-related genes and chlorophyll biosynthesis genes were significantly downregulated in yc412 cotyledons.Our results provide new insights into the molecular function of the YSL transporter in plant chloroplast development and chlorophyll synthesis.

Geographical origin identification of winter jujube(Ziziphus jujuba Dongzao')by using multi-element fingerprinting with chemometrics

Winter jujube(Ziziphus jujuba'Dongzao')is greatly appreciated by consumers for its excellent quality,but brand infringement frequently occurs in the market.Here,we first determined a total of 38 elements in 167 winter jujube samples from the main winter jujube producing areas of China by inductively coupled plasma mass spectrometer(ICP-MS).As a result,16 elements(Mg,K,Mn,Cu,Zn,Mo,Ba,Be,As,Se,Cd,Sb,Ce,Er,Tl,and Pb)exhibited significant differences in samples from different producing areas.Supervised linear discriminant analysis(LDA)and orthogonal projection to latent structures discriminant analysis(OPLS-DA)showed better performance in identifying the origin of samples than unsupervised principal component analysis(PCA).LDA and OPLS-DA had a mean identification accuracy of 87.84 and 94.64%in the testing set,respectively.By using the multilayer perceptron(MLP)and C5.0,the prediction accuracy of the models could reach 96.36 and 91.06%,respectively.Based on the above four chemometric methods,Cd,Tl,Mo and Se were selected as the main variables and principal markers for the origin identification of winter jujube.Overall,this study demonstrates that it is practical and precise to identify the origin of winter jujube through multi-element fingerprint analysis with chemometrics,and may also provide reference for establishing the origin traceability system of other fruits.

Integrated genomic and transcriptomic analysis reveals genes associated with plant height of foxtail millet

Foxtail millet(Setaria italica)is an important C4 model crop;however,due to its high-density planting and high stature,lodging at the filling stage resulted in a serious reduction in yield and quality.Therefore,it is imperative to identify and deploy the genes controlling foxtail millet plant height.In this study,we used a semi-dwarf line 263A and an elite high-stalk breeding variety,Chuang 29 to construct an F2 population to identify dwarf genes.We performed transcriptome analysis(RNA-seq)using internode tissues sampled at three jointing stages of 263A and Chuang 29,as well as bulk segregant analysis(BSA)on their F2 population.A total of 8918 differentially expressed genes(DEGs)were obtained from RNA-seq analysis,and GO analysis showed that DEGs were enriched in functions such as‘‘gibberellin metabolic process”and‘‘oxidoreductase activity”,which have previously been shown to be associated with plant height.A total 593 mutated genes were screened by BSA-seq method.One hundred and seventy-six out of the 593 mutated genes showed differential expression levels between the two parental lines,and seven genes not only showed differential expression in two or three internode tissues but also showed high genomic variation in coding regions,which indicated they play a crucial role in plant height determination.Among them,we found a gibberellin biosynthesis related GA20 oxidase gene(Seita.5G404900),which had a single-base at the third exon,leading to the frameshift mutation at 263A.Cleaved amplified polymorphic sequence assay and association analysis proved the single-base in Seita.5G404900 co-segregated with dwarf phenotype in two independent F2 populations planted in entirely different environments.Taken together,the candidate genes identified in this study will help to elucidate the genetic basis of foxtail millet plant height,and the molecular marker will be useful for marker-assisted dwarf breeding.

Transcriptome Analysis of oserf922 Mutants Reveals New Insights into Rice Blast Resistance

Rice blast disease,caused by fungal Magnaporthe oryzae,severely threatens food security.The susceptibility(S) genes have emerged to support pathogenesis,and disruption of S gene usually confers increased resistance to multiple pathogen isolates.Rice ethylene response factor gene Os ERF922 is a potential S gene in blast disease.However,how Os ERF922 negatively regulates resistance against M.oryzae is still unknown.

Effects of a decade of organic fertilizer substitution on vegetable yield and soil phosphorus pools, phosphatase activities, and the microbial community in a greenhouse vegetable production system

Partial substitution of chemical fertilizers by organic amendments is adopted widely for promoting the availability of soil phosphorus(P)in agricultural production.However,few studies have comprehensively evaluated the effects of longterm organic substitution on soil P availability and microbial activity in greenhouse vegetable fields.A 10-year(2009–2019)field experiment was carried out to investigate the impacts of organic fertilizer substitution on soil P pools,phosphatase activities and the microbial community,and identify factors that regulate these soil P transformation characteristics.Four treatments included 100%chemical N fertilizer(4 CN),50%substitution of chemical N by manure(2 CN+2 MN),straw(2 CN+2 SN),and combined manure with straw(2 CN+1 MN+1 SN).Compared with the 4 CN treatment,organic substitution treatments increased celery and tomato yields by 6.9-13.8%and 8.6-18.1%,respectively,with the highest yields being in the 2 CN+1 MN+1 SN treatment.After 10 years of fertilization,organic substitution treatments reduced total P and inorganic P accumulation,increased the concentrations of available P,organic P,and microbial biomass P,and promoted phosphatase activities(alkaline and acid phosphomonoesterase,phosphodiesterase,and phytase)and microbial growth in comparison with the 4 CN treatment.Further,organic substitution treatments significantly increased soil C/P,and the partial least squares path model(PLS-PM)revealed that the soil C/P ratio directly and significantly affected phosphatase activities and the microbial biomass and positively influenced soil P pools and vegetable yield.Partial least squares(PLS)regression demonstrated that arbuscular mycorrhizal fungi positively affected phosphatase activities.Our results suggest that organic fertilizer substitution can promote soil P transformation and availability.Combining manure with straw was more effective than applying these materials separately for developing sustainable P management practices.

Draft genome sequence of cauliflower(Brassica oleracea L.var.botrytis)provides new insights into the C genome in Brassica species

Cauliflower is an important variety of Brassica oleracea and is planted worldwide.Here,the high-quality genome sequence of cauliflower was reported.The assembled cauliflower genome was 584.60 Mb in size,with a contig N50 of 2.11 Mb,and contained 47,772 genes;56.65%of the genome was composed of repetitive sequences.Among these sequences,long terminal repeats(LTRs)were the most abundant(32.71%of the genome),followed by transposable elements(TEs)(12.62%).Comparative genomic analysis confirmed that after an ancient paleohexaploidy(γ)event,cauliflower underwent two whole-genome duplication(WGD)events shared with Arabidopsis and an additional whole-genome triplication(WGT)event shared with other Brassica species.The present cultivated cauliflower diverged from the ancestral B.oleracea species~3.0 million years ago(Mya).The speciation of cauliflower(~2.0 Mya)was later than that of B.oleracea L.var.capitata(approximately 2.6 Mya)and other Brassica species(over 2.0 Mya).Chromosome no.03 of cauliflower shared the most syntenic blocks with the A,B,and C genomes of Brassica species and its eight other chromosomes,implying that chromosome no.03 might be the most ancient one in the cauliflower genome,which was consistent with the chromosome being inherited from the common ancestor of Brassica species.In addition,2,718 specific genes,228 expanded genes,2 contracted genes,and 1,065 positively selected genes in cauliflower were identified and functionally annotated.These findings provide new insights into the genomic diversity of Brassica species and serve as a valuable reference for molecular breeding of cauliflower.

The puzzle of the missing meat: Food away from home and China's meat statistics

From 1985, an increasing gap has emerged between the official statistical measures of meat production and meat consumption in China, which has raised concerns from many researchers using such data. In this paper we report the results of 428 observations(survey of 107 urban and rural households×4 quarters) from 7 provinces conducted in 2010, and compare them with the official statistical data from the National Bureau of Statistics of China(NBSC). We conclude that the main reason for the discrepancy is due to the underreporting of consumption, which is due mainly to the omission of consumption away from home.

Attraction of bruchid beetles Callosobruchus chinensis (L.) (Coleoptera: Bruchidae) to host plant volatiles

Host-plant volatiles play an important role as cues for herbivores in search of resources, mates and oviposition sites in complex environments. Plant volatile-based attractants can be developed for pest monitoring and control. Previously, we indicated that mated female adults of Callosobruchus chinensis showed choice preference behavior toward 2-hexenal and benzaldehyde. Our objective here was to investigate the synergistic effect of host-derived attractive volatiles in attracting C. chinensis under laboratory and field conditions in Shanxi Province, China. We hypothesized that the ratio and concentration of volatiles derived from Vigna radiata play critical roles for C. chinensis in locating this host. Therefore, we collected and identified the volatiles of mungbean by using headspace collection and GC-MS. The effectiveness of different ratios and concentrations of two compounds (2-hexenal and benzaldehyde) that elicit C. chinensis searching behavior were examined in Y-tube olfactometer assays. The combination of 300 μg μL^–1 2-hexenal and 180 μg μL^–1 benzaldehyde loadings exhibited a synergistic effect on attracting C. chinensis (82.35%). Compared to control traps, the adults were significantly attracted to traps baited with blends, and more attraction to females than males was found for blend traps in the field experiments. Our results suggest that blends of this specific concentration and ratio of benzaldehyde and 2-hexenal can be used in traps as attractants for C. chinensis monitoring and control in the field.

Impacts of Four Invasive Asteraceae on Soil Physico-Chemical Properties and AM Fungi Community

Invasive Asteraceae are an important group of plants and might have universal impacts on invaded ecosystems. However, research data on the ecological impacts of specific plants are still lacking. Chromolaena odorata, Ageratina adenophora, Flaveria bidentis, and Mikania micrantha are four typical alien Asteraceae in China. The involvement of soil biota, in particular, the community structure of arbuscular mycorrhizal (AM) fungi in their invasion process was tested in present research. It was found that invasion by the four Asteraceae changed, to different extents, the nutrient levels in soils. Invasion by C. odorata, A. adenophora, and F. bidentis followed common patterns: 1) decreasing pH value;2) increasing the soil AM fungal diversity and species richness. Invasion by all four Asteraceae tested increased nitrogen pools and accelerated nitrogen fluxes with a decrease in the phosphorus pool, especially available phosphorus. It was suggested that mycorrhizal symbiosis might partially promote successful invasion of these Asteraceae by affecting the metabolism of phosphorus in soil. The impacts on soil ecosystems of these Asteraceae tested were also species-specific, and different invasion strategies were exhibited.

Tracking and Monitoring Leaf Development, Coupling Law and Regulation Techniques during Flowering Period of Hybrid Foxtail Millet (<i>Setaria italica</i>(L.) P. Beauv.) Parental Lines

The determining factor of Setaria italica (L.) P. Beauv. is the coupling of its flowering stage and outcrossing rate which leads to low and unstable seed yields in self-pollinated foxtail millet hybrids and thereby limits their large-scale application. In this study, Datong 27, Datong 29 and gu 83 were screened and identified through meticulous observations of their pollination habitats. High exposure rate, degree of exposure and plump of stigma are good factors to accept foreign pollen. Datong 27 and Datong 29 have some additional characteristics, such as long filaments and exposed and full anthers that contain a large amount of pollen. We transformed into a series of stigma-exposed and plump sterile lines that easily accepted exotic pollen. New restorer lines with anthers that were full of powder and exhibited quick recovery, which improved the parental lines’ heterosexual characteristics. By tracking and monitoring the leaf development of the new sterile and restorer lines, a coupling law of leaf development was determined and a series of flowering control measures were formulated. These factors ensured that the parental lines encounter one another during the flowering stage. By utilizing fertilizer and water, the vitality of the female stigma, amount of powder scattered and powder loosening time were prolonged, which increased hybrid seed yields from 1500 to 3000 kg/hm2. These findings were helpful in resolving the technical problems of seed production that restricted the propagation of foxtail millet hybrids and supporting future large-scale applications.

Genetic Diversity and Classification of Chinese Elite Foxtail Millet [Setaria italica (L.) P. Beauv.] Revealed by Acid-PAGE Prolamin

Arid and semi-arid regions of China account for more than half of the country. Because of drought resistance and high nutritive value, elite foxtail millet (Setaria Italica (L.) P. Beauv.) is one of the most important cereal crops in China. Evaluation of germplasm and genetic diversity of foxtail millet is still in its infancy, but prolamin could play an important role as a protein marker. To investigate the genetic diversity and population structure of foxtail millet from different ecological zones of China, 90 accessions of foxtail millet were collected from three major ecological areas: North, Northwest, and Northeast China. The prolamin contents were examined by acid polyacrylamide gel electrophoresis (acid-PAGE). Five to twenty-two prolamin bands appeared in tested varieties, of which were polymorphic, so prolamin patterns of foxtail millet varieties can be used in variety identification and evaluation. Structure analysis identified six groups, which matches their pedigree information but not their geographic origins. This indicated a high degree (87.78%) of consistency with a phylogenetic classification based on SSR. The results showed prolamin banding patterns were an effective method for analyzing foxtail millet genetic variability.

Effect of Multi-parameter Environmental Factors on Cucumber Leaf Surface Wetness

In this study, artificial leaf resistance was used to simulate leaf wetness. Specific to the solar greenhouse environment in Tianjin, microclimate monitoring equipment was installed for the collection of temperature group and humidity group data, as well as solar radiation and leaf wetness in the greenhouse. In order to reduce the complexity of multivariate factor prediction and ensure the richness of selected data types, correlation analysis was made to the 2 groups of data, screening 5 000 groups of data, including the humidity group data RH, RH_(20), RH_(40), temperature group data T, T_(20), T_(40), and solar radiation W. The data were then analyzed by principal component analysis, screening out 4 groups of principal components to show the leaf wetness index.

Research and Experiment on Performance Stability of Facility Green-house Environmental Monitoring System

At present,the safety and stability of most facility greenhouse environment monitoring systems are seldom considered. In order to improve the stability of data transmission in environment and prevent the system failure caused by the fault of coordinator,a mechanism based on Zigbee coordinator to improve the stability of the whole system is proposed to ensure the security of wireless data transmission. Finally,the system is tested,and the results show that the system can effectively ensure the fault-free transmission of collected environmental data.

Overview of Quality Standardization for Traditional Chinese Medicine

This paper presentsan overview of the standardizationprocess for TraditionalChinese Medicine (TCM)Quality,and reports on the latestresearch projects.It also highlightsthe roles of quality andmodernization in this standardization.

Combining nitrogen effects and metabolomics to reveal the response mechanisms to nitrogen stress and the potential for nitrogen reduction in maize

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.

Utilizing modified ubi1 introns to enhance exogenous gene expression in maize(Zea mays L.) and rice(Oryza sativa L.)

The phenomenon of intron-mediated enhancement(IME) was discovered in 1990 based on the observation that plant introns can stimulate gene expression,particularly in monocots.However,the intrinsic mechanism of IME remains unclear because many studies have yielded various results depending on the promoter,reporter gene,flanking sequences of the intron,and target cell or tissue.In this study,the effect of the first intron of the maize ubiquitin gene(ubi1 intron) was investigated by changing insertion sites,deleting specific regions and mutating individual motifs in maize(Zea mays L.)and rice(Oryza sativa L.) using ubi1 intron-containing GUS(β-glucuronidase) constructs.In maize callus,the integration of the full-length ubi1 intron into the GUS coding sequence at the +13,+115 and +513 positions by particle bombardment increased GUS activity approximately five-,four- and two-fold,respectively.Eight truncated ubi1 introns in the pSG(13i)N construct significantly influenced GUS gene expression to different degrees in transient assays.Notably,the 3' region deletions significantly reduced the IME effect,whereas a 142-nt deletion,pSG(13i-P5)N,in the 5' region caused a 1.5-fold enhancement relative to pSG(13i)N.Furthermore,four site-directed mutageneses were performed in pSG(13i-P5)N;these constructs resulted in the up-regulation of GUS gene expression to different levels.The most effective modified ubi1 intron,pSG(13i-M4)N,was further evaluated and proved in rice using transient experiments.In addition,the sequences flanking the GUS insertion significantly influenced the IME effect of the vectors that were constructed.The modified ubi1 intron had the potential application on crop genetic engineering.

Long-term straw addition promotes moderately labile phosphorus formation, decreasing phosphorus downward migration and loss in greenhouse vegetable soil

Phosphorus(P) leaching is a major problem in greenhouse vegetable production with excessive P fertilizer application. Substitution of inorganic P fertilizer with organic fertilizer is considered a potential strategy to reduce leaching, but the effect of organic material addition on soil P transformation and leaching loss remains unclear. The X-ray absorption nearedge structure(XANES) spectroscopy technique can determine P speciation at the molecular level. Here, we integrated XANES and chemical methods to explore P speciation and transformation in a 10-year field experiment with four treatments: 100% chemical fertilizer(4 CN), 50% chemical N and 50% manure N(2CN+2MN), 50% chemical N and 50% straw N(2CN+2SN), and 50% chemical N and 25% manure N plus 25% straw N(2CN+2 MSN). Compared with the 4 CN treatment, the organic substitution treatments increased the content of labile P by 13.7–54.2% in the 0–40 cm soil layers, with newberyite and brushite being the main constituents of the labile P. Organic substitution treatments decreased the stable P content;hydroxyapatite was the main species and showed an increasing trend with increasing soil depth. Straw addition(2CN+2SN and 2CN+2 MSN) resulted in a higher moderately labile P content and a lower labile P content in the subsoil(60–100 cm). Moreover, straw addition significantly reduced the concentrations and amounts of total P, dissolved inorganic P(DIP), and particulate P in leachate. DIP was the main form transferred by leaching and co-migrated with dissolved organic carbon. Partial least squares path modeling revealed that straw addition decreased P leaching by decreasing labile P and increasing moderately labile P in the subsoil. Overall, straw addition is beneficial for developing sustainable P management strategies due to increasing labile P in the upper soil layer for the utilization of plants, and decreasing P migration and leaching.

The Gray Mold Spore Detection of Cucumber Based on Microscopic Image and Deep Learning

Rapid and accurate detection of pathogen spores is an important step to achieve early diagnosis of diseases in precision agriculture.Traditional detection methods are time-consuming,laborious,and subjective,and image processing methods mainly rely on manually designed features that are difficult to cope with pathogen spore detection in complex scenes.Therefore,an MG-YOLO detection algorithm(Multi-head self-attention and Ghost-optimized YOLO)is proposed to detect gray mold spores rapidly.Firstly,Multi-head self-attention is introduced in the backbone to capture the global information of the pathogen spores.Secondly,we combine weighted Bidirectional Feature Pyramid Network(BiFPN)to fuse multiscale features of different layers.Then,a lightweight network is used to construct GhostCSP to optimize the neck part.Cucumber gray mold spores are used as the study object.The experimental results show that the improved MG-YOLO model achieves an accuracy of 0.983 for detecting gray mold spores and takes 0.009 s per image,which is significantly better than the state-of-the-art model.The visualization of the detection results shows that MG-YOLO effectively solves the detection of spores in blurred,small targets,multimorphology,and high-density scenes.Meanwhile,compared with the YOLOv5 model,the detection accuracy of the improved model is improved by 6.8%.It can meet the demand for high-precision detection of spores and provides a novel method to enhance the objectivity of pathogen spore detection.

Insights into the regulation of C-repeat binding factors in plant cold signaling

Cold temperatures, a major abiotic stress,threaten the growth and development of plants, worldwide. To cope with this adverse environmental cue, plants from temperate climates have evolved an array of sophisticated mechanisms to acclimate to cold periods,increasing their ability to tolerate freezing stress. Over the last decade, significant progress has been made in determining the molecular mechanisms underpinning cold acclimation, including following the identification of several pivotal components, including candidates for cold sensors, protein kinases, and transcription factors. With these developments, we have a better understanding of the CBF-dependent cold-signaling pathway. In this review,we summarize recent progress made in elucidating the cold-signaling pathways, especially the C-repeat binding factor-dependent pathway, and describe the regulatory function of the crucial components of plant cold signaling.We also discuss the unsolved questions that should be the focus of future work.

Current Soil Nutrient Status of Intensively Managed Greenhouses

Nine districts covering the main greenhouse vegetable areas in Tianjin Municipality of the North China Plain were selected for the soil investigation in 2010 to survey the current soil nutrient status (soil available N, P and K), acidification and salinization due to excessive input of fertilizers in greenhouses in Tianjin. The study showed that, in particular, soil available P content increased with the age of greenhouses. In contrast, our results did not reveal higher K accumulation and lowered pH in the greenhouse soils compared with cultivation in open fields. Over-fertilization, causing high NO 3 accumulation, most likely resulted in salinity problems in the greenhouses. Ninety percent of the investigated greenhouse soils had electrical conductivity values of saturated paste extracts of 2-10 d S m-1 , which might affect the yields of vegetable crops like green bean, pepper, cabbage, carrot, eggplant, lettuce, spinach, celery, cucumber and tomato. The findings of our survey of the current fertility and salinity problems in greenhouse soils suggest that there is an urgent need to improve the farmers' practices and strategies in fertilization management in greenhouses of China. Because education and the agricultural technical extension services may play a more important role in avoiding overuse of fertilizers, we suggest that current nutrient management practices should be improved in the near future through training of local farmers in farmers' schools and through strengthening the agricultural extension services with practical techniques.