Knockdown of the atypical protein kinase genes GhABC1K2-A05 and GhABC1K12-A07 make cotton more sensitive to salt and PEG stress

Activity of bc1 complex kinase(ABC1K)is an atypical protein kinase(aPK)that plays a crucial role in plant mitochondrial and plastid stress responses,but little is known about the responses of ABC1Ks to stress in cotton(Gossypium spp.).Here,we identified 40 ABC1Ks in upland cotton(Gossypium hirsutum L.)and found that the Gh ABC1Ks were unevenly distributed across 17 chromosomes.The GhABC1K family members included 35 paralogous gene pairs and were expanded by segmental duplication.The GhABC1K promoter sequences contained diverse cis-acting regulatory elements relevant to hormone or stress responses.The qRT-PCR results revealed that most Gh ABC1Ks were upregulated by exposure to different stresses.Gh ABC1K2-A05 and Gh ABC1K12-A07 expression levels were upregulated by at least three stress treatments.These genes were further functionally characterized by virus-induced gene silencing(VIGS).Compared with the controls,the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced cotton lines exhibited higher malondialdehyde(MDA)contents,lower catalase(CAT),peroxidase(POD)and superoxide dismutase(SOD)activities and reduced chlorophyll and soluble sugar contents under NaCl and PEG stress.In addition,the expression levels of six stress marker genes(Gh DREB2A,Gh SOS1,Gh CIPK6,Gh SOS2,Gh WRKY33,and Gh RD29A)were significantly downregulated after stress in the Gh ABC1K2-A05-and Gh ABC1K12-A07-silenced lines.The results indicate that knockdown of Gh ABC1K2-A05 and Gh ABC1K12-A07 make cotton more sensitive to salt and PEG stress.These findings can provide valuable information for intensive studies of Gh ABC1Ks in the responses and resistance of cotton to abiotic stresses.

Simultaneous determination of five plant hormones in cotton leaves using QuEChERS combined with HPLC‒MS/MS

Background Plant hormones profoundly influence cotton growth,development,and responses to various stresses.Therefore,there is a pressing need for an efficient assay to quantify these hormones in cotton.In this groundbreaking study,we have established QuEChERS-HPLC‒MS/MS method,for the simultaneous detection of multiple plant hormones in cotton leaves,allowing the analysis and quantification of five key plant hormones.Results Sample extraction and purification employed 0.1%acetic acid in methanol and C18 for optimal recovery of plant hormones.The method applied to cotton demonstrated excellent linearity across a concentration range of 0.05–1 mg・L−1,with linear regression coefficients exceeding 0.99.The limits of quantification(LOQs)were 20μg・kg−1 for GA3 and 5μg・kg−1 for the other four plant hormones.Recovery rates for the five plant hormones matrix spiked at levels of 5,10,100,and 1000μg・kg−1 were in the range of 79.07%to 98.97%,with intraday relative standard deviations(RSDs)ranging from 2.11%to 8.47%.The method was successfully employed to analyze and quantify the five analytes in cotton leaves treated with plant growth regulators.Conclusion The study demonstrates that the method is well-suited for the determination of five plant hormones in cotton.It exhibits excellent selectivity and sensitivity in detecting field samples,thus serving as a robust tool for indepth research into cotton physiology.

A glycine-rich nuclear effector VdCE51 of Verticillium dahliae suppresses plant immune responses by inhibiting the accumulation of GhTRXH2

Verticillium dahliae is an important soil-borne fungal pathogen that causes great yield losses in many cash crops.Effectors of this fungus are known to regulate plant immunity but the mechanism much remains unclear.A glycine-rich nuclear effector,VdCE51,was able to suppress immune responses in tobacco against Botrytis cinerea and Sclerotinia sclerotiorum.This effector was a required factor for full virulence of V.dahliae,and its nuclear localization was a requisite for suppressing plant immunity.The thioredoxin GhTRXH2,identified as a positive regulator of plant immunity,was a host target of VdCE51.Our findings show a virulence regulating mechanism whereby the secreted nuclear effector VdCE51 interferes with the transcription of PR genes,and the SA signaling pathway by inhibiting the accumulation of GhTRXH2,thus suppressing plant immunity.

Morphological characteristics,developmental dynamics,and gene temporal expressions across various development stages of Aphis gossypii sexual female

Background Aphis gossypii(Hemiptera:Aphididae)is a worldwide polyphagous phloem-feeding agricultural pest,and it can produce offspring by sexual or asexual reproduction.Compared with dozens of generations by parthenogenesis,sexual reproduction is performed in only one generation within one year,and little is known about the sexual reproduction of A.gossypii.In this study,sexual females of A.gossypii were successfully obtained through a previously established induction platform,and the morphological characteristics,developmental dynamics,and temporal gene expression were examined.Subsequently,signaling pathways potentially involved in regulating the growth,development,and reproduction of sexual females were investigated.Results The morphological observation showed that from the 1st instar nymph to adult,sexual females exhibited a gradually deepened body color,an enlarged body size,longer antennae with a blackened end,and obviously protruding cauda(in adulthood).The anatomy found that the ovaries of sexual females developed rapidly from the 2^(nd)instar nymph,and the embedded oocytes matured in adulthood.In addition,time-course transcriptome analysis revealed that gene expression profiles across the development of sexual females fell into 9 clusters with distinct patterns,in which gene expression levels in clusters 1,5,and 8 peaked at the 2^(nd)instar nymphal stage with the largest number of up-regulated genes,suggesting that the 2^(nd)instar nymph was an important ovary development period.Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis revealed that a large number of genes in the sexual female adult were enriched in the TGF-beta signaling pathway and Forkhead box O(FoxO)signaling pathway,highlighting their important role in sexual female adult development and reproduction.Conclusion The morphological changes of the sexual female at each developmental stage were revealed for the first time.In addition,time-course transcriptomic analyses suggest genes enriched in the TGF-beta signaling pathway and FoxO signaling pathway probably contribute to regulating the development and oocyte maturation of sexual females.Overall,these findings will facilitate the regulating mechanism research in the growth and development of sexual females by providing candidate genes.

Monitoring the little fire ant,Wasmannia auropunctata(Roger 1863),in the early stage of its invasion in China:Predicting its geographical distribution pattern under climate change

Invasive alien ants(IAAs)are among the most aggressive,competitive,and widespread invasive alien species(IAS)worldwide.Wasmannia auropunctata,the greatest IAAs threat in the Pacific region and listed in“100 of the world’s worst IAS”,has established itself in many countries and on islands worldwide.Wild populations of W.auropunctata were recently reported in southeastern China,representing a tremendous potential threat to China’s agricultural,economic,environmental,public health,and social well-being.Estimating the potential geographical distribution(PGD)of W.auropunctata in China can illustrate areas that may potentially face invasion risk.Therefore,based on the global distribution records of W.auropunctata and bioclimatic variables,we predicted the geographical distribution pattern of W.auropunctata in China under the effects of climate change using an ensemble model(EM).Our findings showed that artificial neural network(ANN),flexible discriminant analysis(FDA),gradient boosting model(GBM),Random Forest(RF)were more accurate than categorical regression tree analysis(CTA),generalized linear model(GLM),maximum entropy model(MaxEnt)and surface distance envelope(SRE).The mean TSS values of ANN,FDA,GBM,and RF were 0.820,0.810,0.843,and 0.857,respectively,and the mean AUC values were 0.946,0.954,0.968,and 0.979,respectively.The mean TSS and AUC values of EM were 0.882 and 0.972,respectively,indicating that the prediction results with EM were more reliable than those with the single model.The PGD of W.auropunctata in China is mainly located in southern China under current and future climate change.Under climate change,the PGD of W.auropunctata in China will expand to higher-latitude areas.The annual temperature range(bio7)and mean temperature of the warmest quarter(bio10)were the most significant variables affecting the PGD of W.auropunctata in China.The PGD of W.auropunctata in China was mainly attributed to temperature variables,such as the annual temperature range(bio7)and the mean temperature of the warmest quarter(bio10).The populations of W.auropunctata in southern China have broad potential invasion areas.Developing strategies for the early warning,monitoring,prevention,and control of W.auropunctata in southern China requires more attention.

Thelytokous Diglyphus wani:A more promising biological control agent against agromyzid leafminers than its arrhenotokous counterpart

Diglyphus wani(Hymenoptera:Eulophidae)is a dominant parasitoid that attacks agromyzid leafminers.Two reproductive types occur in D.wani:arrhenotoky(in which virgin females produce only male offspring;and virgin females mate with males to produce bisexual offspring)and thelytoky(in which virgin females produce female offspring).As a potential biological control agent,exploring the differences in the relevant biological parameters of both strains is necessary.In this study,comparisons between the two strains of D.wani were performed by evaluating the life table and host-killing rate.The thelytokous strain exhibited significantly better life table parameters than its arrhenotokous counterpart.Higher values for the intrinsic rate of increase,finite rate of increase,net reproductive rate,and fecundity were found in the thelytokous strain.The thelytokous strain also performed better than the arrhenotokous strain in terms of net parasitism,host-feeding,host-stinging,and total host-killing rates.Thus,populations of the thelytokous strain could grow fast and kill more hosts.In conclusion,the thelytokous strain of D.wani may be the more promising biological agent against agromyzid leafminers compared to its arrhenotokous counterpart.Also,since the thelytokous strain of D.wani is only known to produce females,it should be given priority in future biocontrol applications owing to the cost savings of breeding only females.

Genotypic variation in root morphology, cotton subtending leaf physiology and fiber quality against nitrogen

Background:Nitrogen(N)is important for improving various morphological and physiological processes of cotton but their contribution to fiber quality is still lacking.Aims:The current study aimed to explore the relationship between root morphology,subtending leaf physiology,and fiber quality of contrasting N-efficient cotton genotypes in response to N.Methods:We analyzed the above parameters of CCRI 69(N-efficient)and Xinluzao-30(XLZ-30,N-inefficient)under control(2.5 mmol·L^(-1))and high N(5 mmol·L^(-1))conditions.Results:The results showed that root morphological traits were increased in CCRI-69 under control conditions than high N.Subtending leaf morphology,chlorophyll and carotenoid contents,free amino acids,and soluble proteins were higher under high N as compared with the control.However,soluble sugars,fructose,sucrose contents,and sucrose phosphate synthase were higher under control conditions than high N across the growth stages.Irrespective of the N conditions,all morphological and physiological traits of cotton subtending leaf were higher in CCRI-69 than XLZ-30.Except for fiber uniformity,fiber quality traits like fiber length,strength,micronaire,and elongation were improved under control conditions than high N.Between the genotypes,CCRI-69 had significantly higher fiber length,strength,micronaire,and elongation as compared with XLZ-30.Strong positive correlations were found between root morphology,soluble sugars,sucrose content,and sucrose phosphate synthase activity with fiber quality traits,respectively.Conclusions:These findings suggest that CCRI-69 performed better in terms of growth and fiber quality under relatively low N condition,which will help to reduce fertilizer use,the cost of production,and environmental pollution.

Effect of Indoor Concentrate Feeding vs. Outdoor Grazing on the Expression of Genes Involved in Muscle Growth and Nutrient Content in Japanese Black Steer Muscle

The objective of the present study was to investigate the effect of outdoor grazing on the expression of genes involved in muscle growth and the nutrient contents of skeletal muscle in steers. Ten Japanese Black steers were divided into two groups: grazing (GR) and concentrate (CT) groups. Crude protein, extractable lipid, moisture, fatty acid, cooking loss and Warner Bratzler shear force in muscle tissue were analyzed. The gene expression of myosin heavy chain (MyHC) isoform (2a, 2x and slow), myostatin, follistatin, peroxisome proliferator-activated receptor γ2 (PPARγ2), CCAAT/enhancer binding protein α (C/EBPα), heat shock protein (HSP) 27 and HSP40 in skeletal muscles was evaluated at the end of fattening. Decreases in MyHC-2a and MyHC-2x (fast-twitch fiber type) expression in the longissimus lumborum (LL) muscle were detected in the GR group compared with the CT group;in contrast, an increase in MyHC-slow (slow-twitch fiber type) expression was shown in the GR group. These results suggest that grazing initiated muscle fiber type conversion to slow-twitch from fast-twitch. A decrease in extractable lipid content was observed in the GR group in the LL and semitendinosus (ST) muscles. Crude protein content in the LL muscle in the GR group was higher than in the CT group. MyHC expression in LL muscle in the GR group was greater than in the CT group. A decrease in myostatin and PPARγ2 gene expression was detected in the GR group compared with the CT group in both muscles. Expression of C/EBPα in LL muscle in the GR group was lower than in the CT group. These results suggest that grazing steers at the end of fattening may lead to an increase in protein content and a decrease in fat accumulation in LL and/or ST muscles by regulation of myostatin, MyHC, PPARγ2 and C/EBPα gene expression.

An Improved Open-Top Chamber with Solar-Heated Double Funnels That Can Adapt to All Wind Directions for Simulating Future Global Warming Conditions in Rice Paddy Fields

An Open-Top Chamber with Solar-heated Double Funnels (OTC-SDF2) that uses solar energy as a heating source was developed in this study. Two air entry/exit funnels were connected to the OTC via flat tunnels through which air was warmed by solar radiation. The new apparatus increased the air temperature by approximately 1°C throughout the chamber when more than half the energy of full sunlight was supplied. Although air flow occurs in only two directions, a nearly constant temperature increase was observed for each wind direction. This increase in temperature was stable for 1 mˉs to 4 mˉs wind speeds in every direction. This degree of warming may be adequate for screening high-temperature tolerant plants from medium to weak cultivars. The OTC-SDF2 has the potential to provide moderately high-temperature treatments for screening various cultivars/strains and may be used to evaluate easy, low-cost cropping methods associated with high-temperature stresses.

Small particles,big effects:How nanoparticles can enhance plant growth in favorable and harsh conditions

By 2050,the global population is projected to reach 9 billion,underscoring the imperative for innovative solutions to increase grain yield and enhance food security.Nanotechnology has emerged as a powerful tool,providing unique solutions to this challenge.Nanoparticles(NPs)can improve plant growth and nutrition under normal conditions through their high surface-to-volume ratio and unique physical and chemical properties.Moreover,they can be used to monitor crop health status and augment plant resilience against abiotic stresses(such as salinity,drought,heavy metals,and extreme temperatures)that endanger global agriculture.Application of NPs can enhance stress tolerance mechanisms in plants,minimizing potential yield losses and underscoring the potential of NPs to raise crop yield and quality.This review highlights the need for a comprehensive exploration of the environmental implications and safety of nanomaterials and provides valuable guidelines for researchers,policymakers,and agricultural practitioners.With thoughtful stewardship,nanotechnology holds immense promise in shaping environmentally sustainable agriculture amid escalating environmental challenges.

Field efficacy of insect pathogen, botanical, and jasmonic acid for the management of wheat midge Sitodiplosis mosellana and the impact on adult parasitoid Macroglenes penetrans populations in spring wheat

The wheat midge, Sitodiplosis mosellana, is a serious pest of wheat worldwide. In North America, management of S. mosellana in spring wheat relies on the timely application of pesticides, based on midge adults levels caught in pheromone traps or seen via field scouting during wheat heading. In this context, biopesticides can be an effective alternative to pesticides for controlling S. mosellana within an Integrated Pest Management program. A field study using insect pathogenic fungus Beauveria bassiana GHA, nematode Steinernema Jeltiae with Barricade polymer gel 1%, pyrethrin, combined formulations of B. bassiana GHA and pyrethrin, Jasmonic acid (JA) and chlorpyrifos (chemical check) was performed to determine to which extent they affect midge larval populations, kernel damage levels, grain yield, and quality, and the impacts on adult parasitoid Macroglenes penetrans populations. The results indicated that biopesticides JA and S. Jeltiae were the most effective in reducing larval populations and kernel damage levels, and produced a higher spring wheat yield when compared to the water control at both study locations (East Valier and North Valier, Montana, USA). Increased test weight in wheat had been recorded with two previous biopesticides at East Valier but not for North Valier, when compared over water control. These results were comparable in efficacy to the chlorpyrifos. This study also suggested that B. bassiana and pyrethrin may work synergistically, as exemplified by lower total larval populations and kernel damage levels when applied together. This study did not demonstrate the effect of any treatments on M. penetrans populations.

Traditional Tea-grass Integrated System in Shizuoka as a GIAHS Site: Values and Conservation——Background of Application and Efforts after Registration

The traditional tea-grass integrated system in Shizuoka is the first example of a Globally Important Agriculture Heritage Systems(GIAHS)site in Japan that was proposed by a local government.In this paper,we report the background and circumstances leading to the recognition of this system,its registration as a GIAHS,and its current use after registration.Although semi-natural grasslands have decreased considerably in Japan,we found unique semi-natural grasslands with no pest insects and a rich biodiversity around the tea fields.These grasslands and the farming method employed are known as'Chagusaba',which was registered as a GIAHS in 2013.However,the registration process for GIAHS was not easy,as many local farmers do not understand the value of their traditional farming methods or the GIAHS honor.After registering Chagusaba as a GIAHS,our main agenda was branding.Traditional farming,with time and effort,produces high-quality tea.As labor saving technologies in agriculture have progressed in Japan,the price of high-quality tea has declined,and consequently,the numbers of farmers performing the inefficient traditional farming methods are decreasing.It is necessary to correctly brand the Chagusaba-grown tea and reflect the value of the traditional farming method in the price.The local government of the Shizuoka Prefecture is currently working on implementing a certification system,which includes scientific evaluation using GPS,and the introduction of biodiversity indicators.