Research and Development of New High-quality Short-season Cotton Varieties and the Industrial Application of High Efficiency Technologies in Shandong Province——Based on the Innovation of Building a Green and Efficient Farming System

Through the analysis of the impact of changes in the cotton planting industry on cotton production structure and benefit in Shandong under the national cotton production pattern in past ten years,based on the guarantee of food security and the overall ecological regional layout of Shandong s agricultural characteristic and advantageous industries,we proposed that through the development of high-quality short-season cotton and the industrial application of high-efficiency technologies,a green and high-efficiency farming system should be established to comprehensively reduce cotton planting costs,improve comparative benefits,and realize green and efficient optimization technologies that unify the stable production of cotton and high-quality cotton supply and the industrial path supported by the new industrial model.It is aimed to speed up the upgrading and transformation of the existing traditional cotton cropping system,reconstruct a ternary structure of grains,economic crops and fodder in the ecological suitable cotton areas and sub-suitable cotton areas in Shandong,and maintain a series of new green and ecological cotton rotation industrial models including the“double security”of the grain and cotton industries,the ecological“double superiority”of "cotton and fodder",and the high-efficiency"double increase"of cotton and garlic,providing a certain foundation for the research on technological integration innovation and industrial structure optimization of the green,efficient and high-quality development and transformation of the cotton industry in Shandong during the"14 th Five-Year Plan"period.

One-time fertilization at first flowering improves lint yield and dry matter partitioning in late planted short-season cotton

Cotton producers have substantially reduced their inputs(labor,nutrients,and management)mainly by adopting a shortseason cropping management that is characterized by late sowing,high density,and reduced fertilization with one-time application at the first bloom stage without lint yield reduction.However,it has been hypothesized that one-time fertilization at an earlier growth stage could be a more effective and economic management practice.A two-year field experiment was conducted by applying five fertilizer one-time fertilization at 0(FT1),5(FT2),10(FT3),15(FT4),and 20(FT5)days after the first flower appeared in the field and one three-split fertilizer application taken as the conventional control(FT6),making six treatments altogether.Cotton growth period,biomass accumulation,yield,and its formation were quantified.The results showed that the one-time fertilization did not affect the cotton growth progress as compared to FT6,however,the total crop cycles for FT3–FT5 were 3 days shorter.FT1 produced the highest cotton lint yield(1396 kg ha–1),which was similar to the FT6 but higher than the other treatments,and could be attributed to more bolls per unit area and higher lint percentage.Cotton yield was positively correlated with cotton plant biomass accumulated.FT1 had both the highest average(VT)(193.7 kg ha–1 d–1)and the highest maximum(VM)(220.9 kg ha–1 d–1)rates during the fast biomass accumulation period.These results suggest that one-time fertilizer application at the first flower stage might be an adjustment that is more effective than at first bloom,and allowed for easier decision making for application date due to non counting of plants with flowers is needed.

Growth,leaf anatomy,and photosynthesis of cotton(Gossypium hirsutum L.)seedlings in response to four light-emitting diodes and high pressure sodium lamp

Background Light is a critical factor in plant growth and development,particularly in controlled environments.Light-emitting diodes(LEDs)have become a reliable alternative to conventional high pressure sodium(HSP)lamps because they are more efficient and versatile in light sources.In contrast to well-known specialized LED light spectra for vegetables,the appropriate LED lights for crops such as cotton remain unknown.Results In this growth chamber study,we selected and compared four LED lights with varying percentages(26.44%–68.68%)of red light(R,600–700 nm),combined with other lights,for their effects on growth,leaf anatomy,and photosynthesis of cotton seedlings,using HSP lamp as a control.The total photosynthetic photon flux density(PPFD)was(215±2)μmol·m-2·s-1 for all LEDs and HSP lamp.The results showed significant differences in all tested parameters among lights,and the percentage of far red(FR,701–780 nm)within the range of 3.03%–11.86%was positively correlated with plant growth(characterized by leaf number and area,plant height,stem diameter,and total biomass),palisade layer thickness,photosynthesis rate(Pn),and stomatal conductance(Gs).The ratio of R/FR(4.445–11.497)negatively influenced the growth of cotton seedlings,and blue light(B)suppressed stem elongation but increased palisade cell length,chlorophyll content,and Pn.Conclusion The LED 2 was superior to other LED lights and HSP lamp.It had the highest ratio of FR within the total PPFD(11.86%)and the lowest ratio of R/FR(4.445).LED 2 may therefore be used to replace HPS lamp under controlled environments for the study of cotton at the seedling stage.

Identification of SSR markers linked to the abscission of cotton boll traits and mining germplasm in Cotton

Background Cotton is an economically important crop.It is crucial to find an effective method to improve cotton yield,and one approach is to decrease the abscission of cotton bolls and buds.However,the lack of knowledge of the genetic and molecular mechanisms underlying cotton boll abscission traits has hindered genetic improvements.Results Pearson’s correlation analysis revealed a significant positive correlation between boll abscission rates 1(AR1)and boll abscission rates 2(AR2).A genome-wide association study was conducted on 145 loci that exhibited high polymorphism and were uniformly distributed across 26 chromosomes(pair).The study revealed 18,46,and 62 markers that were significantly associated with boll abscission,fiber quality,and yield traits(P<0.05),explaining 1.75%–7.13%,1.16%–9.58%,and 1.40%–5.44%of the phenotypic variation,respectively.Notably,the marker MON_SHIN-1584b was associated with the cotton boll abscission trait,whereas MON_CGR5732a was associated with cotton boll abscission and fiber quality traits.Thirteen of the marker loci identified in this study had been previously reported.Based on phenotypic effects,six typical cultivars with elite alleles related to cotton boll abscission,fiber quality,and yield traits were identified.These cultivars hold great promise for widespread utilization in breeding programs.Conclusions These results lay the foundation for understanding the molecular regulatory mechanism of cotton boll abscission and provide data for the future improvement of cotton breeding.

Journal of Cotton Research Annual Awards

Hangdling Editor of the Year 2023.Journal of Cotton Research editorial office launches the award of Handling editors of the year to recognize handling editors with the most processed manuscripts in Journal of Cotton Research.Here are the Top 4 Handling editors of 2023:DONG Hezhong,Institute of Industrial Crops,Shandong Academy of Agricultural Sciences,China YANG Guozheng,College of Plant Science&Technology,Huazhong Agricultural University,China ZHANG Lizhen,College of Resources and Environmental Sciences,China Agricultural University,China ZHOU Zhiguo,College of Agriculture,Nanjing Agricultural University,China.

Cotton pads-derived carbon materials/reduced graphene oxide modified with polypyrrole for electrode of supercapacitors

This study investigates the influence of electropolymerization conditions on the deposition of polypyrrole(PPy)onto cotton-derived carbon fiber(CF)modified with reduced graphene oxide(rGO)for supercapacitors applications using an experimental/theorical approach.The surface modification of CF by rGO and/or by PPy electrodeposited at 10,25 and 50 mV s^(-1) was thoroughly examined physicochemical and electrochemically.Composite electrodes comprising CF-rGo-PPy,synthesized via electropolymerization at 25 mV s^(-1),demonstrated a remarkable increase in capacitance,showcasing~742 F g^(-1) compared to 153 F g^(-1) for CF.SEM,N_(2)-surface area,XPS,and TD-DFT approach revealed that the higher capacitance observed in CF-rGo-PPy electrodes underscores the influence of morphology and charged nitrogen species on the electrochemical performance of these modified electrodes.Notably,this electrode material achieves a specific capacitance retention of~96%of their initial capacitance after 10000 cycles at 0.5 A g^(-1) measured in a two-electrodes cell configuration.This work also discusses the influence of the scan rate used for pyrrole electropolymerization on the pseudocapacitance contribution of PPy and its possible effect on the porosity of the material.These results highlight the importance of appropriate electropolymerization conditions that allow obtaining the synergistic effect between CF,rGO and PPy.

Alternate cotton-peanut intercropping:a new approach to increasing productivity and minimizing environmental impact

Recent publications have highlighted the development of an alternate cotton-peanut intercropping as a novel strat-egy to enhance agricultural productivity.In this article,we provide an overview of the progress made in the alternate cotton-peanut intercropping,specifically focusing on its yield benefits,environmental impacts,and the underlying mechanisms.In addition,we advocate for future investigations into the selection or development of appropriate crop varieties and agricultural equipment,pest management options,and the mechanisms of root-canopy interactions.This review is intended to provide a valuable reference for understanding and adopting an alternate intercropping system for sustainable cotton production.

Correction:Identification of SSR markers linked to the abscission of cotton boll traits and mining germplasm in Cotton

Correction:J Cotton Res 7,20(2024)https://doi.org/10.1186/s42397-024-00180-3 Following publication of the original article(Shui et al.2024),the author found 5 errors in the published article.1.One of the author’s name has been corrected from Gou Chunping to Guo Chunping.2.The reference(Zhao SQ.2016)in Table 2 has been updated to:Zhao SQ.Analysis on the major gene and multigene mixed inheritance and QTL mapping for early maturity traits in upland cotton.Chin Acad Agric Sci.2016.https://doi.org/10.3969/j.issn.201600501.(in Chinese with English abstract).3.In’Results’part,’Phenotype analysis of 238 cotton boll abscission among cotton accessions’paragraph,the phenotype analysis of AR1 ranging from 19.27%–63.79%,which was wrongly written as 19.27%-63.97%.4.The‘2018KRL’is modified to‘2018KEL’in Table 1.

Strigolactones modulate cotton fiber elongation and secondary cell wall thickening

Cotton is one of the most important economic crops in the world,and it is a major source of fiber in the textile industry.Strigolactones(SLs)are a class of carotenoid-derived plant hormones involved in many processes of plant growth and development,although the functions of SL in fiber development remain largely unknown.Here,we found that the endogenous SLs were significantly higher in fibers at 20 days post-anthesis(DPA).Exogenous SLs significantly increased fiber length and cell wall thickness.Furthermore,we cloned three key SL biosynthetic genes,namely GhD27,GhMAX3,and GhMAX4,which were highly expressed in fibers,and subcellular localization analyses revealed that GhD27,GhMAX3,and GhMAX4 were localized in the chloroplast.The exogenous expression of GhD27,GhMAX3,and GhMAX4 complemented the physiological phenotypes of d27,max3,and max4 mutations in Arabidopsis,respectively.Knockdown of GhD27,GhMAX3,and GhMAX4 in cotton resulted in increased numbers of axillary buds and leaves,reduced fiber length,and significantly reduced fiber thickness.These findings revealed that SLs participate in plant growth,fiber elongation,and secondary cell wall formation in cotton.These results provide new and effective genetic resources for improving cotton fiber yield and plant architecture.

Enhancing cotton resilience to challenging climates through genetic modifications

Cotton is one of the most important fiber crops that plays a vital role in the textile industry.Its production has been unstable over the years due to climate change induced biotic stresses such as insects,diseases,and weeds,as well as abiotic stresses including drought,salinity,heat,and cold.Traditional breeding methods have been used to breed climate resilient cotton,but it requires a considerable amount of time to enhance crop tolerance to insect pests and changing climatic conditions.A promising strategy for improving tolerance against these stresses is genetic engineering.This review article discusses the role of genetic engineering in cotton improvement.The essential concepts and techniques include genome editing via clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein 9(CRISPR-Cas9),overexpression of target genes,downregulation using RNA interference(RNAi),and virus-induced gene silencing(VIGS).Notably,the Agrobacterium-mediated transformation has made significant contributions to using these techniques for obtaining stable transgenic plants.

Barley chitinase genes expression revamp resistance against whitefly (Bemisia Tabaci) in transgenic cotton (Gossypium hirsutum L.)

Background Chitinase is an enzyme that hydrolyzes chitin,a major component of the exoskeleton of insects,including plant pests like whiteflies.The present study aimed to investigate the expression of chemically synthesized barley ch1 and chi2 genes in cotton(Gossypium hirsutum)through Agrobacterium-mediated transformation.Fifty-five putative transgenic cotton plants were obtained,out of which fifteen plants successfully survived and were shifted to the field.Using gene-specific primers,amplification of 447 bp and 401 bp fragments confirmed the presence of the ch1 and chi2 genes in five transgenic cotton plants of the T0 generation.These five plants were further evalu-ated for their mRNA expression levels.The T0 transgenic cotton plants with the highest mRNA expression level and better yield performance in field,were selected to raise their subsequent progenies.Results The T1 cotton plants showed the highest mRNA expression levels of 3.5-fold in P10(2)for the ch1 gene and 3.7-fold in P2(1)for the chi2 gene.Fluorescent in situ hybridization(FISH)confirmed a single copy number of ch1 and chi2(hemizygous)on chromosome no.6.Furthermore,the efficacy of transgenes on whitefly was evaluated through an insect bioassay,where after 96 h of infestation,mortality rates of whitefly were calculated to be 78%–80%in transgenic cotton plants.The number of eggs on transgenic cotton plants were calculated to be 0.1%–0.12 per plant compared with the non-transgenic plants where egg number was calculated to be 0.90–1.00 per plant.Conclusion Based on these findings,it can be concluded that the chemically synthesized barley chitinase genes(ch1 and chi2)have the potential to be effective against insects with chitin exoskeletons,including whiteflies.The transgenic cotton plants expressing these genes showed increased resistance to whiteflies,resulting in reduced egg numbers and higher mortality rates.

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.

Apical meristem transcriptome analysis identifies a role for the blue light receptor gene GhFKF1 in cotton architecture development

Cotton architecture is determined by the differentiation fate transition of axillary meristem(AM),and influences cotton yield and the efficiency of mechanized harvesting.We observed that the initiation of flowering primordium was earlier in early-maturing than that in late-maturing cultivars during the differentiation and development of AM.The RNA-Seq and expression level analyses showed that genes FLAVIN BINDING,KELCH REPEAT,F-BOX1(GhFKF1),and GIGANTEA(GhGI)were in response to circadian rhythms,and involved in the regulation of cotton flowering.The gene structure,predicted protein structure,and motif content analyses showed that in Arabidopsis,cotton,rapseed,and soybean,proteins GhFKF1 and GhGI were functionally conserved and share evolutionary origins.Compared to the wild type,in GhFKF1 mutants that were created by the CRISPR/Cas9 system,the initiation of branch primordium was inhibited.Conversely,the knocking out of GhGI increased the number of AM differentiating into flower primordium,and there were much more lateral branch differentiation and development.Besides,we investigated that proteins GhFKF1 and GhGI can interact with each other.These results suggest that GhFKF1 and GhGI are key regulators of cotton architecture development,and may collaborate to regulate the differentiation fate transition of AM,ultimately influencing plant architecture.We describe a strategy for using the CRISPR/Cas9 system to increase cotton adaptation and productivity by optimizing plant architecture.

Increasing root-lower characteristics improves drought tolerance in cotton cultivars at the seedling stage

Drought is an important abiotic stress factor in cotton production.The root system architecture(RSA)of cotton shows high plasticity which can alleviate drought-related stress under drought stress(DS)conditions;however,this alleviation is cultivar dependent.Therefore,this study estimated the genetic variability of RSA in cotton under DS.Using the paper-based growth system,we assessed the RSA variability in 80 cotton cultivars at the seedling stage,with 0 and10%polyethylene glycol 6000(PEG6000)as the control(CK)and DS treatment,respectively.An analysis of 23 aboveground and root traits in the 80 cotton cultivars revealed different responses to DS.On the 10th day after DS treatment,the degree of variation in the RSA traits under DS(5–55%)was greater than that of CK(5–49%).The 80 cultivars were divided into drought-tolerant cultivars(group 1),intermediate drought-tolerant cultivars(group 2),and drought-sensitive cultivars(group 3)based on their comprehensive evaluation values of drought resistance.Under DS,the root lengthlower,root area-lower,root volume-lower,and root length density-lower were significantly reduced by 63,71,76,and 4%in the drought-sensitive cultivars compared to CK.Notably,the drought-tolerant cultivars maintained their root lengthlower,root area-lower,root volume-lower,and root length density–lower attributes.Compared to CK,the root diameter(0–2 mm)-lower increased by 21%in group 1 but decreased by 3 and 64%in groups 2 and 3,respectively,under DS.Additionally,the drought-tolerant cultivars displayed a plastic response under DS that was characterized by an increase in the root-lower characteristics.Drought resistance was positively correlated with the root area-lower and root length density-lower.Overall,the RSA of the different cotton cultivars varied greatly under DS.Therefore,important root traits,such as the root-lower traits,provide great insights for exploring whether drought-tolerant cotton cultivars can effectively withstand adverse environments.

Single‑cell RNA sequencing opens a new era for cotton genomic research and gene functional analysis

Single-cell RNA sequencing(scRNA-seq)is one of the most advanced sequencing technologies for studying transcriptome landscape at the single-cell revolution.It provides numerous advantages over traditional RNA-seq.Since it was first used to profile single-cell transcriptome in plants in 2019,it has been extensively employed to perform different research in plants.Recently,scRNA-seq was also quickly adopted by the cotton research community to solve lots of scientific questions which have been never solved.In this comment,we highlighted the significant progress in employing scRNA-seq to cotton genetic and genomic study and its future potential applications.

Genetics of biochemical attributes regulating morpho-physiology of upland cotton under high temperature conditions

Background Cotton is a strategically important fibre crop for global textile industry.It profoundly impacts several countries’industrial and agricultural sectors.Sustainable cotton production is continuously threatened by the unpre-dictable changes in climate,specifically high temperatures.Breeding heat-tolerant,high-yielding cotton cultivars with wide adaptability to be grown in the regions with rising temperatures is one of the primary objectives of modern cotton breeding programmes.Therefore,the main objective of the current study is to figure out the effective breed-ing approach to imparting heat tolerance as well as the judicious utilization of commercially significant and stress-tolerant attributes in cotton breeding.Initially,the two most notable heat-susceptible(FH-115 and NIAB Kiran)and tolerant(IUB-13 and GH-Mubarak)cotton cultivars were spotted to develop filial and backcross populations to accom-plish the preceding study objectives.The heat tolerant cultivars were screened on the basis of various morphological(seed cotton yield per plant,ginning turnout percentage),physiological(pollen viability,cell membrane thermostabil-ity)and biochemical(peroxidase activity,proline content,hydrogen peroxide content)parameters.Results The results clearly exhibited that heat stress consequently had a detrimental impact on every studied plant trait,as revealed by the ability of crossing and their backcross populations to tolerate high temperatures.However,when considering overall yield,biochemical,and physiological traits,the IUB-13×FH-115 cross went over particularly well at both normal and high temperature conditions.Moreover,overall seed cotton yield per plant exhibited a posi-tive correlation with both pollen viability and antioxidant levels(POD activity and proline content).Conclusions Selection from segregation population and criteria involving pollen viability and antioxidant levels concluded to be an effective strategy for the screening of heat-tolerant cotton germplasms.Therefore,understanding acquired from this study can assist breeders identifying traits that should be prioritized in order to develop climate resilient cotton cultivars.

Comparative analysis of SIMILAR to RCD ONE(SRO)family from tetraploid cotton species and their diploid progenitors depict their significance in cotton growth and development

Background SRO(Similar to RCD1)genes family is largely recognized for their importance in the growth,develop-ment,and in responding to environmental stresses.However,genome-wide identification and functional characteri-zation of SRO genes from cotton species have not been reported so far.Results A total of 36 SRO genes were identified from four cotton species.Phylogenetic analysis divided these genes into three groups with distinct structure.Syntenic and chromosomal distribution analysis indicated uneven distribu-tion of GaSRO,GrSRO,GhSRO,and GbSRO genes on A2,D5 genomes,Gh-At,Gh-Dt,Gb-At,and Gb-Dt subgenomes,respectively.Gene duplication analysis revealed the presence of six duplicated gene pairs among GhSRO genes.In promoter analysis,several elements responsive to the growth,development and hormones were found in GhSRO genes,implying gene induction during cotton growth and development.Several miRNAs responsive to plant growth and abiotic stress were predicted to target 12 GhSRO genes.Organ-specific expression profiling demonstrated the roles of GhSRO genes in one or more tissues.In addition,specific expression pattern of some GhSRO genes dur-ing ovule development depicted their involvement in these developmental processes.Conclusion The data presented in this report laid a foundation for understanding the classification and functions of SRO genes in cotton.

The DUF579 proteins GhIRX15s regulate cotton fiber development by interacting with proteins involved in xylan synthesis

Cotton provides the most abundant natural fiber for the textile industry.The mature cotton fiber largely consists of secondary cell walls with the highest proportion of cellulose and a small amount of hemicellulose and lignin.To dissect the roles of hemicellulosic polysaccharides during fiber development,four IRREGULAR XYLEM 15(IRX15)genes,GhIRX15-1/-2/-3/-4,were functionally characterized in cotton.These genes encode DUF579 domain-containing proteins,which are homologs of AtIRX15 involved in xylan biosynthesis.The four GhIRX15 genes were predominantly expressed during fiber secondary wall thickening,and the encoded proteins were localized to the Golgi apparatus.Each GhIRX15 gene could restore the xylan deficient phenotype in the Arabidopsis irx15irx15l double mutant.Silencing of GhIRX15s in cotton resulted in shorter mature fibers with a thinner cell wall and reduced cellulose content as compared to the wild type.Intriguingly,GhIRX15-2 and GhIRX15-4 formed homodimers and heterodimers.In addition,the GhIRX15s showed physical interaction with glycosyltransferases GhGT43C,GhGT47A and GhGT47B,which are responsible for synthesis of the xylan backbone and reducing end sequence.Moreover,the GhIRX15s can form heterocomplexes with enzymes involved in xylan modification and side chain synthesis,such as GhGUX1/2,GhGXM1/2 and GhTBL1.These findings suggest that GhIRX15s participate in fiber xylan biosynthesis and modulate fiber development via forming large multiprotein complexes.

Laboratory evaluation of toxicity of selected insecticides against egg and larval stages of cotton pink bollworm, Pectinophora gossypiella (Saunders) (Lepidoptera: Gelechiidae)

Background The cryptic nature of pink bollworm Pectinophora gossypiella(Saunders)larvae enables its reduced vul-nerability to insecticidal control.Further,the development of resistance against Bacillus thuringiensis(Bt)toxins posed a serious threat to transgenic cotton cultivation.This necessitated determining the critical timing of spray applications on the control effectiveness.This study assessed the influence of egg age(freshly laid vs.three-day-old)and the loca-tion of larvae(directly exposed to the insecticide residues on the boll rind vs.burrowed inside the bolls)on insecticide control efficacy.Results The results revealed a significant decrease in the ovicidal activity of tested insecticides with an increase in the age of eggs from one day old to three days old(paired t-test,P<0.05).The larvae directly exposed to the insec-ticide residues on the boll rind were more susceptible(>80%mortality)than the larvae exposed after they had bur-rowed inside the bolls(<49%mortality).The inhibitory effects of tested insecticides on developmental biology were more pronounced in the experiment on pre-larval release insecticide treatment compared with insecticide treatment given post-larval release and entry inside the bolls.Conclusion Egg age influences the insecticide susceptibility,as does the larval location,directly exposed vs bur-rowed inside the bolls.Older eggs and the larvae that had burrowed inside the green bolls of cotton were relatively less susceptible to the insecticide treatments.The toxic effects of insecticides on egg and larval stages were primar-ily ephemeral.These findings are significant for devising a comprehensive strategy for pink bollworm management on a sustainable basis.

Inhibition of protein degradation increases the Bt protein concentration in Bt cotton

Bacillus thuringiensis(Bt)cotton production is challenged by two main problems,i.e.,the low concentration of Bt protein at the boll setting stage and the lowest insect resistance in bolls among all the cotton plant’s organs.Therefore,increasing the Bt protein concentration at the boll stage,especially in bolls,has become the main goal for increasing insect resistance in cotton.In this study,two protein degradation inhibitors(ethylene diamine tetra acetic acid(EDTA)and leupeptin)were sprayed on the bolls,subtending leaves,and whole cotton plants at the peak flowering stage of two Bt cultivars(medium maturation Sikang 1(SK1))and early maturation Zhongmian 425(ZM425)in 2019 and 2020.The Bt protein content and protein degradation metabolism were assessed.The results showed that the Bt protein concentrations were enhanced by 21.3 to 38.8%and 25.0 to 38.6%in the treated bolls of SK1 and ZM425 respectively,while they were decreased in the subtending leaves of these treated bolls.In the treated leaves,the Bt protein concentrations increased by 7.6 to 23.5%and 11.2 to 14.9%in SK1 and ZM425,respectively.The combined application of EDTA and leupeptin to the whole cotton plant increased the Bt protein concentrations in both bolls and subtending leaves.The Bt protein concentrations in bolls were higher,increasing by 22.5 to 31.0%and 19.6 to 32.5%for SK1 and ZM425,respectively.The organs treated with EDTA or/and leupeptin showed reduced free amino acid contents,protease and peptidase activities and significant enhancements in soluble protein contents.These results indicated that inhibiting protein degradation could improve the protein content,thus increasing the Bt protein concentrations in the bolls or/and leaves of cotton plants.Therefore,the increase in the Bt protein concentration without yield reduction suggested that these two protein degradation inhibitors may be applicable for improving insect resistance in cotton production.