Stomatal dynamics are regulated by leaf hydraulic traits and guard cell anatomy in nine true mangrove species

Stomatal regulation is critical for mangroves to survive in the hyper-saline intertidal zone where water stress is severe and water availability is highly fluctuant.However,very little is known about the stomatal sensitivity to vapour pressure deficit(VPD)in mangroves,and its co-ordination with stomatal morphology and leaf hydraulic traits.We measured the stomatal response to a step increase in VPD in situ,stomatal anatomy,leaf hydraulic vulnerability and pressure-volume traits in nine true mangrove species of five families and collected the data of genome size.We aimed to answer two questions:(1)Does stomatal morphology influence stomatal dynamics in response to a high VPD in mangroves?with a consideration of possible influence of genome size on stomatal morphology;and(2)do leaf hydraulic traits influence stomatal sensitivity to VPD in mangroves?We found that the stomata of mangrove plants were highly sensitive to a step rise in VPD and the stomatal responses were directly affected by stomatal anatomy and hydraulic traits.Smaller,denser stomata was correlated with faster stomatal closure at high VPD across the species of Rhizophoraceae,and stomata size negatively and vein density positively correlated with genome size.Less negative leaf osmotic pressure at the full turgor(πo)was related to higher operating steady-state stomatal conductance(gs);and a higher leaf capacitance(Cleaf)and more embolism resistant leaf xylem were associated with slower stomatal responses to an increase in VPD.In addition,stomatal responsiveness to VPD was indirectly affected by leaf morphological traits,which were affected by site salinity and consequently leaf water status.Our results demonstrate that mangroves display a unique relationship between genome size,stomatal size and vein packing,and that stomatal responsiveness to VPD is regulated by leaf hydraulic traits and stomatal morphology.Our work provides a quantitative framework to better understand of stomatal regulation in mangroves in an environment with high salinity and dynamic water availability.

Extremely thin but very robust:Surprising cryptogam trait combinations at the end of the leaf economics spectrum

Leaf economics spectrum(LES)describes the fundamental trade-offs between leaf structural,chemical,and physiological investments.Generally,structurally robust thick leaves with high leaf dry mass per unit area(LMA)exhibit lower photosynthetic capacity per dry mass(Amass).Paradoxically,“soft and thinleaved”mosses and spikemosses have very low Amass,but due to minute-size foliage elements,their LMA and its components,leaf thickness(LT)and density(LD),have not been systematically estimated.Here,we characterized LES and associated traits in cryptogams in unprecedented details,covering five evolutionarily different lineages.We found that mosses and spikemosses had the lowest LMA and LT values ever measured for terrestrial plants.Across a broad range of species from different lineages,Amass and LD were negatively correlated.In contrast,Amass was only related to LMA when LMA was greater than 14 g cm^(-2).In fact,low Amass reflected high LD and cell wall thickness in the studied cryptogams.We conclude that evolutionarily old plant lineages attained poorly differentiated,ultrathin mesophyll by increasing LD.Across plant lineages,LD,not LMA,is the trait that represents the trade-off between leaf robustness and physiology in the LES.

Identification, pathogenicity, and fungicide sensitivity of Eutiarosporella dactylidis associated with leaf blight on maize in China

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 2018and 2020 to identify the disease-causing pathogen. Based on morphological features, pathogenicity tests, and multilocus 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.

Characterization of mulberry leaf instant tea and evaluation of its hypolipidemia effect via regulation of intestinal microbiota

The purpose of this study was to characterize mulberry leaf instant tea(MLIT)powder prepared from the'Longsang No.1'(Morus abla L.cv.Longsang 1)mulberry leaves in Heilongjiang Province(China)and assess its obesity-preventing/relieving effects.A total of 174 compounds including quercetin,chlorogenic acid,1-deoxyecomycin(1-DNJ)related to antihyperlipidemia effects were identified from the MLIT powder.MLIT treatment reversed the Lee's index,fat coefficient,and serum biochemical parameters in both the obesity relieving and obesity preventing mice fed with high-fat diet.In the obesity relieving experiment,the relative abundance of Desulfovibrio in mouse feces decreased after both 0.5%and 1%MLIT treatments.In obesity preventing experiments,mouse with different amount of MLIT treatments showed increased relative abundance of Akkermansia,Bifidobacterium and Lactobacillus,while Deferribacteres,Desulfobacterota decreased.The beneficial bacteria in the intestinal tract of mice treated with MLIT increased.This study proved that MLIT had antihyperlipidemia potential via modulating intestinal microbiota in mice.

Real-Time Multiple Guava Leaf Disease Detection from a Single Leaf Using Hybrid Deep Learning Technique

The guava plant has achieved viable significance in subtropics and tropics owing to its flexibility to climatic environments,soil conditions and higher human consumption.It is cultivated in vast areas of Asian and Non-Asian countries,including Pakistan.The guava plant is vulnerable to diseases,specifically the leaves and fruit,which result in massive crop and profitability losses.The existing plant leaf disease detection techniques can detect only one disease from a leaf.However,a single leaf may contain symptoms of multiple diseases.This study has proposed a hybrid deep learning-based framework for the real-time detection of multiple diseases from a single guava leaf in several steps.Firstly,Guava Infected Patches Modified MobileNetV2 and U-Net(GIP-MU-NET)has been proposed to segment the infected guava patches.The proposed model consists of modified MobileNetv2 as an encoder,and the U-Net model’s up-sampling layers are used as a decoder part.Secondly,the Guava Leaf SegmentationModel(GLSM)is proposed to segment the healthy and infected leaves.In the final step,the Guava Multiple Leaf Diseases Detection(GMLDD)model based on the YOLOv5 model detects various diseases from a guava leaf.Two self-collected datasets(the Guava Patches Dataset and the Guava Leaf Diseases Dataset)are used for training and validation.The proposed method detected the various defects,including five distinct classes,i.e.,anthracnose,insect attack,nutrition deficiency,wilt,and healthy.On average,the GIP-MU-Net model achieved 92.41%accuracy,the GLSM gained 83.40%accuracy,whereas the proposed GMLDD technique achieved 73.3%precision,73.1%recall,71.0%mAP@0.5 and 50.3 mAP@0.5:0.95 scores for all the aforesaid classes.

Gene expression profiles in early leaf of rice(Oryza sativa)and foxtail millet(Setaria italica)

Plant anatomy is patterned early during leaf development which suggests studying the spatial–temporal transcriptomes of primordia will help identify critical regulative and functional genes.We successfully isolated the leaf primordia tissues from the C3grass rice and the C4grass foxtail millet by laser capture microdissection(LCM)and studied the gene expression throughout leaf developmental stages.Our data analysis uncovered the conserved expression patterns of certain gene clusters both in rice and foxtail millet during leaf development.We revealed genes and transcription factors involved in vein formation,stomatal development,and suberin accumulation.We identified 79 candidate genes associated with functional regulation of C4anatomy formation.Screening phenotype of the candidate genes revealed that knock-out of a putative polar auxin transport related gene NAL1 resulted significantly reduced veinal space in rice leaf.Our present work provides a foundation for future analyses of genes with novel functions in grasses and their role in leaf development,in particular the role in leaves with a contrasting C3vs.C4biosynthetic pathway.

Leaf Morphology Genes SRL1 and RENL1 Co-Regulate Cellulose Synthesis and Affect Rice Drought Tolerance

The morphological development of rice(Oryza sativa L.)leaves is closely related to plant architecture,physiological activities,and resistance.However,it is unclear whether there is a co-regulatory relationship between the morphological development of leaves and adaptation to drought environment.In this study,a drought-sensitive,roll-enhanced,and narrow-leaf mutant(renl1)was induced from a semi-rolled leaf mutant(srl1)by ethyl methane sulfonate(EMS),which was obtained from Nipponbare(NPB)through EMS.Map-based cloning and functional validation showed that RENL1 encodes a cellulose synthase,allelic to NRL1/OsCLSD4.The RENL1 mutation resulted in reduced vascular bundles,vesicular cells,cellulose,and hemicellulose contents in cell walls,diminishing the water-holding capacity of leaves.In addition,the root system of the renl1 mutant was poorly developed and its ability to scavenge reactive oxygen species(ROS)was decreased,leading to an increase in ROS after drought stress.Meanwhile,genetic results showed that RENL1 and SRL1 synergistically regulated cell wall components.Our results revealed a theoretical basis for further elucidating the molecular regulation mechanism of cellulose on rice drought tolerance,and provided a new genetic resource for enhancing the synergistic regulation network of plant type and stress resistance,thereby realizing simultaneous improvement of multiple traits in rice.

ZmbZIP27 regulates nitrogen-mediated leaf angle by modulating lignin deposition in maize

In grain crops such as maize(Zea mays),leaf angle(LA)is a key agronomic trait affecting light interception and thus planting density and yield.Nitrogen(N)affects LA in plants,but we lack a good understanding of how N regulates LA.Here,we report that N deficiency enhanced lignin deposition in the ligular region of maize seedlings.In situ hybridization showed that the bZIP transcription factor gene ZmbZIP27 is mainly expressed in the phloem of maize vascular bundles.Under N-sufficient conditions,transgenic maize overexpressing ZmbZIP27 showed significantly smaller LA compared with wild type(WT).By contrast,zmbzip27_(ems)mutant showed larger LA under both N-deficient and N-sufficient conditions compared with WT.Overexpression of ZmbZIP27 enhanced lignin deposition in the ligular region of maize in the field.We further demonstrated that ZmbZIP27 could directly bind the promoters of the microRNA genes ZmMIR528a and ZmMIR528b and negatively regulate the expression levels of ZmmiR528.ZmmiR528 knockdown transgenic maize displayed erect architecture in the field by increasing lignin content in the ligular region of maize.Taken together,these results indicate that ZmbZIP27 regulates N-mediated LA size by regulating the expression of ZmmiR528 and modulating lignin deposition in maize.

A novel transcription factor FnMYB4 regulates pigments metabolism of yellow leaf mutants in Fragaria nilgerrensis

The strawberry species Fragaria nilgerrensis Schlechtendal ex J.Gay,renowned for its distinctive white,fragrant peach-like fruits and strong disease resistance,is an exceptional research material.In a previous study,an ethyl methane sulfonate(EMS)mutant library was established for this species,resulting in various yellow leaf mutants.Leaf yellowing materials are not only the ideal materials for basic studies on photosynthesis mechanism,chloroplast development,and molecular regulation of various pigments,but also have important utilization value in ornamental plants breeding.The present study focused on four distinct yellow leaf mutants:mottled yellow leaf(MO),yellow green leaf(YG),light green leaf(LG),and buddha light leaf(BU).The results revealed that the flavonoid content and carotenoid-to-chlorophyll ratio exhibited a significant increase among these mutants,while experiencing a significant decrease in chlorophyll and carotenoid contents compared to the wild type(WT).To clarify the regulatory mechanisms and network relationships underlying these mutants,the RNA-seq and weighted gene coexpression network(WGCNA)analyses were employed.The results showed flavonoid metabolism pathway was enriched both in MO and YG mutants,while the chlorophyll biosynthesis pathway and carotenoid degradation pathway were only enriched in MO and YG mutants,respectively.Subsequently,key structural genes and transcription factors were identified on metabolic pathways of three pigments through correlation analyses and quantitative experiments.Furthermore,a R2R3-MYB transcription factor,FnMYB4,was confirmed to be positively correlated with flavonoid synthesis through transient overexpression,virus-induced gene silencing(VIGS),and RNA interference(RNAi),accompanying by reoccurrence and attenuation of mutant phenotype.Finally,dual-luciferase(LUC)and yeast one-hybrid assays confirmed the binding of FnMYB4 to the FnFLS and FnF3H promoters,indicating that FnMYB4 positively regulates flavonoid synthesis.In addition,correlation analyses suggested that FnMYB4 also might be involved in chlorophyll and carotenoid metabolisms.These findings demonstrated the pivotal regulatory role of FnMYB4 in strawberry leaf coloration.

The anthocyanin formation of purple leaf is associated with the activation of LfiHY5 and LfiMYB75 in crape myrtle

Purple-leafed plants not only have a higher resistance to biotic and abiotic stresses,but also have higher ornamental value.Anthocyanins are vital for leaf color formation,growth and development of purple leaves.However,the molecular mechanism underlying purple leaf formation in Lagerstroemia indica remains unclear.Metabolomic and transcriptomic analysis of purple-leafed cultivar‘Ebony Embers’and greenleafed cultivar‘Arapahoe’showed that the high expression of anthocyanin structure genes induced hyperaccumulation of cyanidin and pelargonidin derivatives,making the leaves purple.LfiHY5,LfiMYB75 and LfibHLH1 were identified using correlation analysis and weighted gene co-expression network analysis.In‘Arapahoe’‘Ebony Embers’population,LfiHY5 and LfiMYB75 showed significant positive correlation with leaf anthocyanin content.Transient expression of LfiMYB75 and LfiHY5 in tobacco and purple-leafed crape myrtle indicated that the two genes activated anthocyanin synthesis.Yeast two-hybrid analysis showed that LfiMYB75 and LfibHLH1 could form a complex that enhanced anthocyanin synthesis.Yeast monohybrid and dual-luciferase assays confirmed that LfiHY5 activated the expression of LfiMYB75,to activate the transcription of anthocyanin structural genes LfiCHS and LfiANS.Moreover,there were three alleles of LfiHY5 in crape myrtle,and the different sequences had different activation effects on LfiMYB75.In conclusion,the results showed that LfiHY5 led to upregulate the transcription of LfiMYB75,and LfiMYB75 formed a complex with LfibHLH1,which increased the transcription level of LfiCHS and LfiANS to affect anthocyanin synthesis in crape myrtle.

Synergistic effects of planting density and nitrogen fertilization on chlorophyll degradation and leaf senescence after silking in maize

Regulating planting density and nitrogen(N)fertilization could delay chlorophyll(Chl)degradation and leaf senescence in maize cultivars.This study measured changes in ear leaf green area(GLA_(ear)),Chl content,the activities of Chl a-degrading enzymes after silking,and the post-silking dry matter accumulation and grain yield under multiple planting densities and N fertilization rates.The dynamic change of GLA_(ear)after silking fitted to the logistic model,and the GLA_(ear) duration and the GLAearat 42 d after silking were affected mainly by the duration of the initial senescence period(T_(1))which was a key factor of the leaf senescence.The average chlorophyllase(CLH)activity was 8.3 times higher than pheophytinase activity and contributed most to the Chl content,indicating that CLH is a key enzyme for degrading Chl a in maize.Increasing density increased the CLH activity and decreased the Chl content,T1,GLAear,and GLA_(ear) duration.Under high density,appropriate N application reduced CLH activity,increased Chl content,prolonged T1,alleviated high-density-induced leaf senescence,and increased post-silking dry matter accumulation and grain yield.

Metabolic profiles and morphological characteristics of leaf tips among different sweet potato(Ipomoea batatas Lam.)varieties

Sweet potato leaf tips have high nutritional value,and exploring the differences in the metabolic profiles of leaf tips among different sweet potato varieties can provide information to improve their qualities.In this study,a UPLC-Q-Exactive Orbitrap/MS-based untargeted metabolomics method was used to evaluate the metabolites in leaf tips of 32 sweet potato varieties.Three varieties with distinct overall metabolic profiles(A01,A02,and A03),two varieties with distinct profiles of phenolic acids(A20 and A18),and three varieties with distinct profiles of flavonoids(A05,A12,and A16)were identified.In addition,a total of 163 and 29 differentially expressed metabolites correlated with the color and leaf shape of sweet potato leaf tips,respectively,were identified through morphological characterization.Group comparison analysis of the phenotypic traits and a metabolite-phenotypic trait correlation analysis indicated that the color differences of sweet potato leaf tips were markedly associated with flavonoids.Also,the level of polyphenols was correlated with the leaf shape of sweet potato leaf tips,with lobed leaf types having higher levels of polyphenols than the entire leaf types.The findings on the metabolic profiles and differentially expressed metabolites associated with the morphology of sweet potato leaf tips can provide useful information for breeding sweet potato varieties with higher nutritional value.

Rice melatonin deficiency causes premature leaf senescence via DNA methylation regulation

In a study of DNA methylation changes in melatonin-deficient rice mutants,mutant plants showed premature leaf senescence during grain-filling and reduced grain yield.Melatonin deficiency led to transcriptional reprogramming,especially of genes involved in chlorophyll and carbon metabolism,redox regulation,and transcriptional regulation,during dark-induced leaf senescence.Hypomethylation of mCG and mCHG in the melatonin-deficient rice mutants was associated with the expression change of both protein-coding genes and transposable element-related genes.Changes in gene expression and DNA methylation in the melatonin-deficient mutants were compensated by exogenous application of melatonin.A decreased S-adenosyl-L-methionine level may have contributed to the DNA methylation variations in rice mutants of melatonin deficiency under dark conditions.

A growth-regulating factor 7(GRF7)-mediated gene regulatory network promotes leaf growth and expansion in sugarcane

Knowledge of the function of growth-regulating factors(GRFs)in sugarcane(Saccharum officinarum and S.spontaneum)growth and development could assist breeders in selecting desirable plant architectures.However,limited information about GRFs is available in Saccharum due to their polyploidy.In this study,22 GRFs were identified in the two species and their conserved domains,gene structures,chromosome location,and synteny were characterized.GRF7 expression varied among tissues and responded to diurnal rhythm.SsGRF7-YFP was localized preferentially in the nucleus and appears to act as a transcriptional cofactor.SsGRF7 positively regulated the size and length of rice leaves,possibly by regulating cell size and plant hormones.Of seven potential transcription factors binding to the SsGRF7 promoter in S.spontaneum,four showed positive expression patterns,and two showed negative expression patterns relative to SsGRF7.

Rice ONAC016 promotes leaf senescence through abscisic acid signaling pathway involving OsNAP

Senescence-induced NAC(senNAC)TFs play a crucial role in senescence during the final stage of leaf development.In this study,we identified a rice senNAC,ONAC016,which functions as a positive regulator of leaf senescence.The expression of ONAC016 increased rapidly in rice leaves during the progression of dark-induced and natural senescence.The onac016-1 knockout mutant showed a delayed leaf yellowing phenotype,whereas the overexpression of ONAC016 accelerated leaf senescence.Notably,ONAC016 expression was upregulated by abscisic acid(ABA),and thus detached leaves of the onac016-1 mutant remained green much longer under ABA treatment.Quantitative RT-PCR analysis showed that ONAC016 upregulates the genes associated with chlorophyll degradation,senescence,and ABA signaling.Yeast one-hybrid and dual-luciferase assays revealed that ONAC016 binds directly to the promoter regions of OsNAP,a key gene involved in chlorophyll degradation and ABA-induced senescence.Taken together,these results suggest that ONAC016 plays an important role in promoting leaf senescence through the ABA signaling pathway involving OsNAP.

Litter production and leaf nutrient concentration and remobilization in response to climate seasonality in the central Amazon

Litterfall is the largest source of nutrients to for-est soils of tropical rainforests.However,variability in lit-terfall production,nutrient remobilization,and changes in leaf nutrient concentration with climate seasonality remain largely unknown for the central Amazon.This study meas-ured litterfall production,leaf nutrient remobilization,and leaf area index on a forest plateau in the central Amazon.Litterfall was measured at monthly intervals during 2014,while nitrogen,phosphorus,potassium,calcium and mag-nesium concentrations of leaf litter and canopy leaves were measured in the dry and rainy seasons,and remobilization rates determined.Leaf area index was also recorded in the dry and rainy seasons.Monthly litterfall varied from 33.2(in the rainy season)to 87.6 g m^(-2) in the dry season,while leaf area index increased slightly in the rainy season.Climatic seasonality had no effect on concentrations of nitrogen,calcium,and magnesium,whereas phosphorous and potassium responded to rainfall seasonality oppositely.While phosphorous increased,potassium decreased during the dry season.Over seasons,nitrogen,potassium,and phosphorous decreased in leaf litter;calcium increased in leaf litter,while magnesium remained unaffected with leaf aging.Regardless,the five nutrients had similar remobilization rates over the year.The absence of climate seasonality on nutrient remobilization suggests that the current length of the dry season does not alter nutrient remobilization rates but this may change as dry periods become more prolonged in the future due to climate change.

Ethylene accelerates maize leaf senescence in response to nitrogen deficiency by regulating chlorophyll metabolism and autophagy

Leaf senescence is an orderly and highly coordinated process,and finely regulated by ethylene and nitrogen(N),ultimately affecting grain yield and nitrogen-use efficiency(NUE).However,the underlying regulatory mechanisms on the crosstalk between ethylene-and N-regulated leaf senescence remain a mystery in maize.In this study,ethylene biosynthesis gene ZmACS7 overexpressing(OE-ZmACS7)plants were used to study the role of ethylene regulating leaf senescence in response to N deficiency,and they exhibited the premature leaf senescence accompanied by increased ethylene release,decreased chlorophyll content and F_v/F_m ratio,and accelerated chloroplast degradation.Then,we investigated the dynamics changes of transcriptome reprogramming underlying ethylene-accelerated leaf senescence in response to N deficiency.The differentially expressed genes(DEGs)involved in chlorophyll biosynthesis were significantly down-regulated,while DEGs involved in chlorophyll degradation and autophagy processes were significantly up-regulated,especially in OE-ZmACS7 plants in response to N deficiency.A gene regulatory network(GRN)was predicted during ethylene-accelerated leaf senescence in response to N deficiency.Three transcription factors(TFs)ZmHSF4,Zmb HLH106,and ZmEREB147 were identified as the key regulatory genes,which targeted chlorophyll biosynthesis gene ZmLES22,chlorophyll degradation gene ZmNYC1,and autophagy-related gene ZmATG5,respectively.Furthermore,ethylene signaling key genes might be located upstream of these TFs,generating the signaling cascade networks during ethylene-accelerated leaf senescence in response to N deficiency.Collectively,these findings improve our molecular knowledge of ethylene-accelerated maize leaf senescence in response to N deficiency,which is promising to improve NUE by manipulating the progress of leaf senescence in maize.

Fine mapping of two recessive genes TaFLA1 and TaSPL8 controlling flag leaf angle in bread wheat

Flag leaf angle is one of the key target traits in high yield wheat breeding.A smaller flag leaf angle reduces shading and enables plants to grow at a higher density,which increases yield.Here we identified a mutant,je0407,with an 84.34%-89.35%smaller flag leaf angle compared with the wild type.The mutant also had an abnormal lamina joint and no ligule or auricle.Genetic analysis indicated that the ligule was controlled by two recessive genes,which were mapped to chromosomes 2AS and 2DL.The mutant allele on chromosome 2AS was named Tafla1b,and it was fine mapped to a 1 Mb physical interval.The mutant allele on chr.2DL was identified as Taspl8b,a novel allele of TaSPL8 with a missense mutation in the second exon,which was used to develop a cleaved amplified polymorphic sequence marker.F3 and F4 lines derived from crosses between Jing411 and je0407 were genotyped to investigate interactions between the Tafla1b and Taspl8b alleles.Plants with the Tafla1b/Taspl8a genotype had 58.41%-82.76%smaller flag leaf angles,6.4%-24.9%shorter spikes,and a greater spikelet density(0.382 more spikelets per cm)compared with the wild type.Plants with the Tafla1a/Taspl8b genotype had 52.62%-82.24%smaller flag leaf angles and no differences in plant height or spikelet density compared with the wild type.Tafla1b/Taspl8b plants produced erect leaves with an abnormal lamina joint.The two alleles had dosage effects on ligule formation and flag leaf angle,but no significant effect on thousand-grain weight.The mutant alleles provide novel resources for improvement of wheat plant architecture.

Gene expression,transcription factor binding and histone modification predict leaf adaxial-abaxial polarity related genes

Leaf adaxial-abaxial(ad-abaxial)polarity is crucial for leaf morphology and function,but the genetic machinery governing this process remains unclear.To uncover critical genes involved in leaf ad-abaxial patterning,we applied a combination of in silico prediction using machine learning(ML)and experimental analysis.A Random Forest model was trained using genes known to influence ad-abaxial polarity as ground truth.Gene expression data from various tissues and conditions as well as promoter regulation data derived from transcription factor chromatin immunoprecipitation sequencing(ChIP-seq)was used as input,enabling the prediction of novel ad-abaxial polarity-related genes and additional transcription factors.Parallel to this,available and newly-obtained transcriptome data enabled us to identify genes differentially expressed across leaf ad-abaxial sides.Based on these analyses,we obtained a set of 111 novel genes which are involved in leaf ad-abaxial specialization.To explore implications for vegetable crop breeding,we examined the conservation of expression patterns between Arabidopsis and Brassica rapa using single-cell transcriptomics.The results demonstrated the utility of our computational approach for predicting candidate genes in crop species.Our findings expand the understanding of the genetic networks governing leaf ad-abaxial differentiation in agriculturally important vegetables,enhancing comprehension of natural variation impacting leaf morphology and development,with demonstrable breeding applications.

Structural elucidation of mulberry leaf oligosaccharide and its selective promotion of gut microbiota to alleviate type 2 diabetes mellitus

Two oligosaccharide fractions(MLO 2-1 and 2-2)were purified from enzymatic hydrolysate of mulberry leaf polysaccharide.The results of simulated digestion showed that MLO 2-2 was a digestible oligosaccharide,which could be degraded by human digestive juice;while MLO 2-1 possessed the non-digestible property in the upper gastrointestinal tract and performed the function by regulating the gut microbiota.Hence,MLO 2-1 was selected for the further analysis.The structure of MLO 2-1 was elucidated as follow:α-T-Glcp-(1→3)-β-Glcp-(1→5)-α-Araf-(1→5)-α-Araf-1→5)-α-Araf-(1→3)-α-(2-OAc)-Glcp-1.The in vitro fecal fermentation results showed that MLO 2-1 could modulate the composition of gut microbiota.Meanwhile,MLO 2-1 was effectively metabolized by fecal bacteria to produce lactate and short chain fatty acids,especially acetate and butyrate.The specific metabolic pathways of MLO 2-1 by gut microbiota were further illuminated.Gut microbiota analysis revealed that MLO 2-1 selectively promoted the growth of Ligilactobacillus murinus,a commensal bacterium presented a reduced level in T2DM mice.Animal experiments indicated that MLO 2-1 and L.murinus exhibited hypoglycemic activities.These results demonstrated that MLO 2-1 might alleviate T2DM by selectively accelerating the proliferation of L.murinus.