Genetic dissection and origin of pleiotropic loci underlying multilevel fiber quality traits in upland cotton(Gossypium hirsutum L.)

Cotton fiber quality is a persistent concern that determines planting benefits and the quality of finished textile products.However,the limitations of measurement instruments have hindered the accurate evaluation of some important fiber characteristics such as fiber maturity,fineness,and neps,which in turn has impeded the genetic improvement and industrial utilization of cotton fiber.Here,12 single fiber quality traits were measured using Advanced Fiber Information System(AFIS)equipment among 383 accessions of upland cotton(Gossypium hirsutum L.).In addition,eight conventional fiber quality traits were assessed by the High Volume Instrument(HVI)System.Genome-wide association study(GWAS),linkage disequilibrium(LD)block genotyping and functional identification were conducted sequentially to uncover the associated elite loci and candidate genes of fiber quality traits.As a result,the previously reported pleiotropic locus FL_D11 regulating fiber length-related traits was identified in this study.More importantly,three novel pleiotropic loci(FM_A03,FF_A05,and FN_A07)regulating fiber maturity,fineness and neps,respectively,were detected based on AFIS traits.Numerous highly promising candidate genes were screened out by integrating RNA-seq and qRT-PCR analyses,including the reported GhKRP6 for fiber length,the newly identified GhMAP8 for maturity and GhDFR for fineness.The origin and evolutionary analysis of pleiotropic loci indicated that the selection pressure on FL_D11,FM_A03 and FF_A05 increased as the breeding period approached the present and the origins of FM_A03 and FF_A05 were traced back to cotton landraces.These findings reveal the genetic basis underlying fiber quality and provide insight into the genetic improvement and textile utilization of fiber in G.hirsutum.

Identification of candidate genes for early-maturity traits by combining BSA-seq and QTL mapping in upland cotton(Gossypium hirsutum L.)

Cotton breeding for the development of early-maturing varieties is an effective way to improve multiple cropping indexes and alleviate the conflict between grains and cotton in the cultivated fields in China.In the present study,we aimed to identify upland cotton quantitative trait loci(QTLs)and candidate genes related to early-maturity traits,including whole growth period(WGP),flowering timing(FT),node of the first fruiting branch(NFFB),height of the node of the first fruiting branch(HNFFB),and plant height(PH).An early-maturing variety,CCRI50,and a latematuring variety,Guoxinmian 11,were crossed to obtain biparental populations.These populations were used to map QTLs for the early-maturity traits for two years(2020 and 2021).With BSA-seq analysis based on the data of population 2020,the candidate regions related to early maturity were found to be located on chromosome D03.We then developed 22 polymorphic insertions or deletions(InDel)markers to further narrow down the candidate regions,resulting in the detection of five and four QTLs in the 2020 and 2021 populations,respectively.According to the results of QTL mapping,two candidate regions(InDel_G286-InDel_G144 and InDel_G24-InDel_G43)were detected.In these regions,three genes(GH_D03G0451,GH_D03G0649,and GH_D03G1180)have nonsynonymous mutations in their exons and one gene(GH_D03G0450)has SNP variations in the upstream sequence between CCRI50 and Guoxinmian 11.These four genes also showed dominant expression in the floral organs.The expression levels of GH_D03G0451,GH_D03G0649 and GH_D03G1180 were significantly higher in CCRI50 than in Guoxinmian 11 during the bud differentiation stages,while GH_D03G0450 showed the opposite trend.Further functional verification of GH_D03G0451 indicated that the GH_D03G0451-silenced plants showed a delay in the flowering time.The results suggest that these are the candidate genes for cotton early maturity,and they may be used for breeding early-maturity cotton varieties.

Mining elite loci and candidate genes for root morphology-related traits at the seedling stage by genome-wide association studies in upland cotton(Gossypium hirsutum L.)

Root system architecture plays an essential role in water and nutrient acquisition in plants,and it is significantly involved in plant adaptations to various environmental stresses.In this study,a panel of 242 cotton accessions was collected to investigate six root morphological traits at the seedling stage,including main root length(MRL),root fresh weight(RFW),total root length(TRL),root surface area(RSA),root volume(RV),and root average diameter(AvgD).The correlation analysis of the six root morphological traits revealed strong positive correlations of TRL with RSA,as well as RV with RSA and AvgD,whereas a significant negative correlation was found between TRL and AvgD.Subsequently,a genome-wide association study(GWAS)was performed using the root phenotypic and genotypic data reported previously for the 242 accessions using 56,010 single nucleotide polymorphisms(SNPs)from the CottonSNP80K array.A total of 41 quantitative trait loci(QTLs)were identified,including nine for MRL,six for RFW,nine for TRL,12 for RSA,12 for RV and two for AvgD.Among them,eight QTLs were repeatedly detected in two or more traits.Integrating these results with a transcriptome analysis,we identified 17 candidate genes with high transcript values of transcripts per million(TPM)≥30 in the roots.Furthermore,we functionally verified the candidate gene GH_D05G2106,which encodes a WPP domain protein 2in root development.A virus-induced gene silencing(VIGS)assay showed that knocking down GH_D05G2106significantly inhibited root development in cotton,indicating its positive role in root system architecture formation.Collectively,these results provide a theoretical basis and candidate genes for future studies on cotton root developmental biology and root-related cotton breeding.

GhWDL3 is involved in the formation and development of fiber cell morphology in upland cotton(Gossypium hirsutum L.)

Background Cotton fiber is a model tissue for studying microtubule-associated proteins(MAPs).The Xklp2(TPX2)proteins that belong to the novel MAPs member mainly participate in the formation and development of microtubule(MT).However,there is a lack of studies concerning the systematic characterization of the TPX2 genes family in cotton.Therefore,the identification and portrayal of G.hirsutum TPX2 genes can provide key targets for molecular manipula-tion in the breeding of cotton fiber improvement.Result In this study,TPX2 family genes were classified into two distinct subclasses TPXLs and MAP genes WAVE DAMP-ENED2-LIKE(WDLs)and quite conservative in quantity.GhWDL3 was significantly up-regulated in 15 days post anthe-sis fibers of ZRI-015(an upland cotton with longer and stronger fiber).GhWDL3 promotes all stem hairs to become straight when overexpressed in Arabidopsis,which may indirectly regulate cotton fiber cell morphology during fiber development.Virus induced gene silencing(VIGS)results showed that GhWDL3 inhibited fiber cell elongation at fiber development periods through regulating the expression of cell wall related genes.Conclusion These results reveal that GhWDL3 regulated cotton fiber cell elongation and provide crucial information for the further investigation in the regulatory mechanisms/networks of cotton fiber length.

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.

Characterization of early maturing elite genotypes based on MTSI and MGIDI indexes:an illustration in upland cotton(Gossypium hirsutum L.)

Background Globally,the cultivation of cotton is constrained by its tendency for extended periods of growth.Early maturity plays a potential role in rainfed-based multiple cropping system especially in the current era of climate change.In the current study,a set of 20 diverse Gossypium hirsutum genotypes were evaluated in two crop seasons with three planting densities and assessed for 11 morphological traits related to early maturity.The study aimed to identify genotype(s)that mature rapidly and accomplish well under diverse environmental conditions based on the two robust multivariate techniques called multi-trait stability index(MTSI)and multi-trait genotype-ideotype distance index(MGIDI).Results MTSI analysis revealed that out of the 20 genotypes,three genotypes,viz.,NNDC-30,A-2,and S-32 accomplished well in terms of early maturity traits in two seasons.Furthermore,three genotypes were selected using MGIDI method for each planting densities with a selection intensity of 15%.The strengths and weaknesses of the genotypes selected based on MGIDI method highlighted that the breeders could focus on developing early-maturing genotypes with specific traits such as days to first flower and boll opening.The selected genotypes exhibited positive genetic gains for traits related to earliness and a successful harvest during the first and second pickings.However,there were negative gains for traits related to flowering and boll opening.Conclusion The study identified three genotypes exhibiting early maturity and accomplished well under different planting densities.The multivariate methods(MTSI and MGIDI)serve as novel approaches for selecting desired genotypes in plant breeding programs,especially across various growing environments.These methods offer exclusive benefits and can easily construe and minimize multicollinearity issues.

Expression analysis of the R2R3-MYB gene family in upland cotton and functional study of GhMYB3D5 in regulating Verticillium wilt resistance

Improving plant resistance to Verticillium wilt(VW),which causes massive losses in Gossypium hirsutum,is a global challenge.Crop plants need to efficiently allocate their limited energy resources to maintain a balance between growth and defense.However,few transcriptional regulators specifically respond to Verticillium dahliae and the underlying mechanism has not been identified in cotton.In this study,we found that the that expression of most R2R3-MYB members in cotton is significantly changed by V.dahliae infection relative to the other MYB types.One novel R2R3-MYB transcription factor(TF)that specifically responds to V.dahliae,GhMYB3D5,was identified.GhMYB3D5 was not expressed in 15 cotton tissues under normal conditions,but it was dramatically induced by V.dahliae stress.We functionally characterized its positive role and underlying mechanism in VW resistance.Upon V.dahliae infection,the up-regulated GhMYB3D5 bound to the GhADH1 promoter and activated GhADH1expression.In addition,GhMYB3D5 physically interacted with GhADH1 and further enhanced the transcriptional activation of GhADH1.Consequently,the transcriptional regulatory module GhMYB3D5-GhADH1 then promoted lignin accumulation by improving the transcriptional levels of genes related to lignin biosynthesis(GhPAL,GhC4H,Gh4CL,and GhPOD/GhLAC)in cotton,thereby enhancing cotton VW resistance.Our results demonstrated that the GhMYB3D5 promotes defense-induced lignin accumulation,which can be regarded as an effective way to orchestrate plant immunity and growth.

Exogenous application of bio-stimulants and growth retardants improve nutrient absorption and fiber quality in upland cotton

Background Natural and synthetic plant growth regulators are essential for plant health,likewise these regulators also play a role in increasing organic production productivity and improving quality and yield stability.In the present study,we have evaluated the effects of foliar applied plant growth regulators,i.e.,moringa leaf extract(MLE)and mepiquat chloride(MC)alone and in combination MC and MLE on the conventional cotton cultivar(CIM 573)and transgenic one(CIM 598).The growth regulators were applied at the start of bloom,45 and 90 days after blooming.Results The application of MC and MLE at 90 days after blooming significantly improved the relative growth rate,net assimilation rate,the number of bolls per plant,and seed cotton yield.Likewise,the combined application of MLE and MC at 90 days after blooming significantly boosted the nitrogen uptake in locules,as well as the phosphorus and potassium uptake in the leaves of both cotton cultivars.The application of MLE alone has considerably improved the nitrogen uptake in leaves,and phosphorus and potassium contents in locules of Bt and conventional cotton cultivars.Similarly,Bt cotton treated with MLE at 90 days after blooming produced significantly higher ginning out turn and oil contents.Treatment in combination(MLE+MC)at 90 days after blooming produced considerably higher micronaire value,fiber strength,and staple length in conventional cultivar.Conclusion The natural growth enhancer,MLE is a rich source of minerals and zeatin,improving the nutrient absorption and quality of cotton fiber in both conventional and Bt cotton cultivars.

Relation Analysis of Several Agronomic Traits and Single Plant Lint Yield in Upland Cotton with High Quality

[ Objective] The purpose was to study the correlation between main agronomic traits and single plant lint yield in upland cotton with high quality. [ Method] Twenty-four upland cotton lines with high quality were analyzed for single plant lint yield and 10 agronomic traits in a randomized, complete block at the agriculture experimental station of JXAU, Nanchang, China in 2007. They were divided into three types （ high, medium, low yield） based on single plant lint yield by Ward＇s method. A total of 11 traits of three types were compared. Correlation a- nalysis and stepwise regression analysis of 10 agronomic traits to single plant lint yield were carried out. [ Result] There existed statistically sig- nificant difference in bolls per plant, ~int percentage, lint index, fruit node numbers, growth period for three yield types. The high yield type ex- hibited the highest bolls per plant, lint percentage and lint index, the lowest fruit node numbers and the shortest growth period. Bolls per plant, boll weight and single plant lint yield were significantly and positively correlated. Fruit node numbers and single plant lint yield were significantly and negatively correlated. Bolls per plant, boll weight, and fruiting position number were the most important factors influencing single plant lint yield of upland cotton lines with high quality. [ Conclusion ] The results will have certain significance for the development of upland cotton variety with high quality.

Obtaining High Pest_resistant Transgenic Upland Cotton Cultivars Carrying cry1Ac3 Gene Driven by Chimeric OM Promoter

Hypocotyl segments from aseptic seedlings of two important cultivars of upland cotton ( Gossypium hirsutum L.) in Northwest China, 'Xinluzao_1', 'Jinmian_7', 'Jinmian_12' and 'Jihe_321' were transformed respectively by two efficient plant expression plasmids pBinMoBc and pBinoBc via Agrobacterium tumefaciens . In pBinMoBc, cry 1Ac3 gene, which encodes the Bt toxin, is under the control of chimeric OM promoter. In pBinoBc, it is under control of CaMV 35S promoter. After co_cultivation with Agrobacterium tumefimpfaciens LBA4404 (containing pBinMoBc or pBinoBc), kanamycin_resistant selection, somatic embryos were induced and regenerated plants were obtained. Then the regenerated plantlets were grafted to untransformed stocks in greenhouse to produce descendants. The integration of cry 1Ac3 gene and its expression in T 2 generation of transgenic cotton plants were confirmed by Southern hybridization and Western blotting. The analyses of insect bioassay indicated that the transgenic plants of both constructions have significant resistance to the larvae of cotton bollworm ( Heliothis armigera ) and that cry 1Ac3 gene driven by chimeric OM promoter could endue T 2 generation cotton with high pest_resistant ability, implicating that it has a profound application in genetic engineering to breed new pest_resistant cotton varieties.

Effects on Agronomic Traits of M_1 by Pollen of Upland Cotton Irradiated by ^(60)Co-γ Ray

[Objective] The aim was to provide reference for research on radiation and breeding of cotton pollen through irradiating common ripe pollen grain of upland cotton by 60Co-γ Ray of varied doses. [Method] Ripe pollen grains of upland cotton were irradiated by 60Co-γ Ray with doses of 5, 10, 15, and 20 Gy, respectively, to learn radiation effect and select appropriate dose. [Result] Most properties of M1 obviously showed variation when dose was over 10 Gy; vitality, growth, and fertility were greatly inhibited when dose was 15 Gy which was almost semi-lethal concentration, and variation species were richest at the same time, which provided materi- als for practical breeding. [Conclusion] 60Co-γ Ray of 15 Gy is more suitable for mutagenesis research on ripe pollen grains of upland cotton.

QTL Mapping of Yield and Yield Components for Elite Hybrid Derived-RILs in Upland Cotton

A population of 180 recombinant inbred lines （RILs） was developed by single seed descended from the cross of high yield Upland cotton （Gossypium hirsutum L.） varieties Zhongmiansuo12 （ZMSI2） and 8891, the two parents of Xiangzamian2 （XZM2）. A genetic linkage map consisting of 132 loci and covering 865.20 cM was constructed using the RIL population chiefly with SSR markers. Yield and yield components were investigated for RILs in three environments in China. The purpose of the present research was to analyze the relationship between yield and its components and to map QTL for yield and yield components in cotton. QTL were tagged with data sets from single environment （separate analysis） and a set of data from means of the three environments （joint analysis）. A total of 34 QTL for yield and yield components were independently detected in three environments, whereas fifteen QTL were found in joint analysis. Notably, a stable lint percentage QTL qLP-A10-1 was detected both in joint analysis and in two environments of separate analysis, which might be of special value for marker-assisted selection. The QTL detected in the present study provide new information on improving yield and yield components. Results of path analysis showed that bolls/plant had the largest contribution to lint yield, which is consistent with the mid-parent heterosis value in F1. Accordingly, in cotton breeding, bolls/plant can be considered first and other yield components measured as a whole to implement variety enhancement and hybrid selection of cotton.

Comparison of the Activity Differences in Two Terminal Oxidases between Upland Cotton and Sea-island Cotton

[Objective] This study was to reveal the differences in two terminal oxdiases,so as to provide theoretical basis for the selection of upland and sea-land hybrid materials. [Methods] Using 7 shares of upland cotton cultivars and 4 shares of sea-land cultivars as experimental materials,the change differences in AAO and PPO activities during whole growth period of the materials tested were studied. [Results] Two terminal oxdiases both in upland and sea-land cotton cultivars assumed similar change laws during whole growth period. AAO activity assumed a low-high trend,and there is a peak at the early stage. PPO activity presented two peaks,and that at the early stage was higher than at the late stage; the first peak is obviously higher than the second for sea-land cultivars,and two peaks are not remarkably different for upland cultivars. [Conclusion] The differences in AAO and PPO activities of sea-land and upland cotton cultivars observed on Jul.4th and Aug.6th,may provide guidance for the selection of upland and sea-land hybrids.

Bioinformatics Analysis of PHYB Gene in Upland Cotton(Gossypium hirsutum)

[Objective] This study was conducted to clarify the biological information of PHYB genes in upland cotton （Gossypium hirsutum）. [Method] Two PHYB genes were identified from the genome database of allotetraploid cotton （G. hirsutum L. acc. TM-1）, and were found to be distributed on subgenomes A10 and D10. And then bioinformatic analysis on these two genes were performed. [Result] The PHYB genes of upland cotton had the same motifs and domains with the PHYB genes in other plant species, and even the number and location of the motifs and domains of these PHYB genes were consistent. The PHYB amino acid sequence alignment and the phylogenetic tree constructed based on PHYB amino acid sequence of these plant species indicated that the two PHYB genes in upland cotton had higher homology and closer evolutionary relationships with cocoa （Theobroma cacao）, but lower similarity to PHYB genes in monocotyledonous plants, such as rice （Oryza saitva） and corn （Zea mays）. The comparison of PHYB gene structure also revealed that plant PHYB gene was more conserved during evolution. The autophosphorylation of dozens of phosphorylation sites in upland cotton PHYB gene may be essential for the functions of phytochromes and plays a significant role in regulating phytochrome-mediated signal transduction pathways. [Conclusion] The results of this paper will provide a theoretical basis for the cloning and functional research of PHYB genes.

Development of a core set of SNP markers for the identifi cation of upland cotton cultivars in China

Considering the advantages of single nucleotide polymorphisms（SNP） in genotyping and variety identification, the first set public SNP markers at Cotton Marker Database（http：//www.cottonmarker.org/） were validated and screened across standard varieties of cotton distinctness, uniformity and stability（DUS） test, aiming to obtain an appropriate set of core SNP markers suitable for upland cotton cultivars in China. A total of 399 out of 1 005 SNPs from 270 loci including 170 insertions-deletions（In Dels） were evaluated for their polymorphisms among 30 standard varieties using Sanger sequencing. As a result, 147 loci were sequenced successfully, 377 SNPs and 49 In Dels markers were obtained. Among the 377 SNP markers, 333 markers（88.3%） were polymorphic between Gossypium hirsutum and G. barbadense, while 164 markers（43.5%） were polymorphic within upland cotton. As for In Del markers, the polymorphic rate is relatively lower than that of SNP both between species and within species. The homozygous DNA locus ratio of 121 SNPs was higher than 86.2% while that of other 43 SNPs was less than 70%. Only 64 SNPs displayed completely homozygous genotypes among all of the detected upland cotton varieties with 100% homozygous DNA locus ratio. At last, a set of 23 pairs of core SNPs were achieved in view of avoidance of linkage, with polymorphism information content（PIC） values varying from 0.21 to 0.38 with an average of 0.28. Genotype characteristics and genetic diversity were analyzed based on the set of core markers, while 40 pairs of core simple-sequence repeats（SSR） primers comprised of 10 sets of four multiplex PCR combinations were also used for analysis based on fluorescence detection system. Comparison results indicated that the genetic diversity level was almost equal, while various varieties were significantly different from each other. Genetic relationship revealed by SSR markers is related to geographic source to a certain extent. Meanwhile clustering results analyzed by SNP markers are more consistent with kinship, which demonstrated that the screen strategy for core SNP marker is effective.

Major Gene Identification and Quantitative Trait Locus Mapping for YieldRelated Traits in Upland Cotton(Gossypium hirsutum L.)

Segregation analysis of the mixed genetic model of major gene plus polygene was used to identify the major genes for cotton yield-related traits using six generations P1, P2, F1, B1, B2, and F2 generated from the cross of Baimian 1 x TM-1. In addition to boll size and seed index, the major genes for the other five traits were detected： one each for seed yield, lint percentage, boll number, lint index; and two for lint yield. Quantitative trait locus/loci （QTL） mapping was performed in the F2 and F2：3 populations of above cross through molecular marker technology, and a total of 50 QTL （26 suggestive and 24 significant） for yield-related traits were detected. Four common QTL were discovered： qLP-3b（F2）/qLP-3（F2：3） and qLP-19b （F2）/qLP-19（F2：3） for lint percentage, qBN-17（F2）/qBN-17（F2：3） for boll number, and qBS-26b（F2）/qBS-26（F2：3） for boll size. Especially, qLP- 3b（Fz）/qLP-3（F2：3）, not only had LOD scores 〉3 but also exceeded the permutation threshold （5.13 and 5.29, respectively）, correspondingly explaining 23.47 and 29.55% of phenotypic variation. This QTL should be considered preferentially in marker assisted selection （MAS）. Segregation analysis and QTL mapping could mutually complement and verify, which provides a theoretical basis for genetic improvement of cotton yield-related traits by using major genes （QTL）.

Characterization of Gh SERK2 and its expression associated with somatic embryogenesis and hormones level in Upland cotton

Somatic embryogenesis （SE） is one of the most important steps during regeneration of cotton, but the molecular mechanism of SE remains unclear. SOMATIC EMBRYOGENSIS RECEPTOR KINASE （SERK） gene is known to function in SE. A homolog GhSERK2 （accession number： JF430801） was cloned from Upland cotton and characterized for its functions in SE. GhSERK2 expressed in different tissues and showed higher expression level in floral organs than vegetative ones with the highest levels in ovule and anther. GhSERK2 expressed during SE with a high level at globular embryos stage. Upon treatment with indole-3-butytic acid （IBA）, the transcription level of GhSERK2 was induced and promoted SE subsequently. A 2-day treatment of 2,4-dichlorophenoxyacetic acid （2,4-D） induced the expression of GhSERK2, but treatments of 2,4-D for longer periods sharply inhibited the GhSERK2 transcription level of embryogenic callus （EC）. The levels of hormones, including 3-indoleacetic acid （IAA）, abscisic acid （ABA）, and brassinosteroid （BR）, were increased in the initial calli induced from the over-expression of GhSERK2 cotton. Our results indicated that GhSERK2 expression was associated with induction of SE and closely related to hormone levels during tissue culture in Upland cotton, and the gene might play an important role in regeneration of cotton.

Improvement of copper tolerance of Arabidopsis by transgenic expression of an allene oxide cyclase gene,GhA OC1, in upland cotton(Gossypium hirsutum L.)

Allene oxide cyclase(AOC, E 5.3.99.6) is an essential enzyme in the jasmonic acid(JA)biosynthetic pathway and mediates a wide range of adaptive responses. In this report, five AOC genes(Gh AOC1–Gh AOC5) were cloned from upland cotton(Gossypium hirsutum L.),sequenced, and characterized. Real-time PCR analysis indicated that the transcripts of Gh AOCs were abundantly expressed in roots and less in fibers, and regulated in cotton plants under methyl jasmonate(Me JA) and Cu Cl2 stresses. To investigate the role of Gh AOC under copper stress, transgenic Arabidopsis plants overexpressing cotton Gh AOC1 under control of the Cauliflower mosaic virus 35S(Ca MV 35S) promoter were generated. Compared to untransformed plants, Gh AOC1-overexpressing Arabidopsis thaliana plants exhibited markedly higher survival rate, shoot fresh weight, shoot dry weight, and photosynthetic efficiency, and reduced cell membrane damage and lipid peroxidation under copper stress.This study provides the first evidence that Gh AOC1 plays an important role in copper stress tolerance.

Transcriptome and QTL analyses reveal candidate genes for fiber quality in Upland cotton

With increasing demand for high-quality cotton,it is desirable to identify genes involved in fiber development for molecular improvement of cotton.In this study,780 differentially expressed genes(DEGs)were identified in developing fibers at 10 days post-anthesis(DPA)in Gossypium hirsutum acc.DH962 and G.hirsutum cv.Jimian 5 using RNA-seq.Of 15 stable QTL for fiber quality identified in the same two parents in previous studies,4,3,6,1,and 1 QTL were associated with fiber length(FL),fiber strength(FS),micronaire(MIC),fiber elongation(FE)and fiber length uniformity ratio(FU),respectively.Integration of DEGs and QTL allowed the identification of 31 genes in 9 QTL regions,of which 25 were highly expressed in fibers based on the transcriptome datasets and 9 were preferentially expressed in different stages of fiber development.Gh_A01G0453(GhDTX19),Gh_D07G1581 and Gh_D04G0942 were expressed specifically in 5 and 10 DPA fibers,with Gh_D04G0942 showing low expression in other tissues except pistil.Gh_D07G1799(GhGAUT9),Gh_D11G0326(GhVPS29),Gh_D11G0333(GhTCP14),and Gh_D11G0334(GhNRP2)were preferentially expressed in 5 or 10 DPA fibers;Gh_A01G0397(GhABCG10)and Gh_D07G1744 were expressed specifically in 20 and 25 DPA fibers.These results suggest candidate genes for molecular improvement of cotton fiber quality.

Development and identification of Verticillium wilt-resistant upland cotton accessions by pyramiding QTL related to resistance

Cotton Verticillium wilt is a serious soil-borne disease that leads to significant losses in fiber yield and quality worldwide. Currently, the most effective way to increase Verticillium wilt resistance is to develop new resistant cotton varieties. Lines 5026 and 60182 are two Verticillium wilt-resistant upland cotton accessions. We previously identified a total of 25 quantitative trait loci（QTLs） related to Verticillium wilt resistance from 5026 and 60182 by assembling segregating populations from hybridization with susceptible parents. In the current study, using 13 microsatellite markers flanking QTLs related to Verticillium wilt resistance, we developed 155 cotton inbred lines by pyramiding different QTLs related to Verticillium wilt resistance from a filial generation produced by crossing 5026 and 60182. By examining each allele＇s effect and performing multiple comparison analysis, we detected four elite QTLs/alleles（q-5/NAU905-2, q-6/NAU2754-2, q-8/NAU3053-1 and q-13/NAU6598-1） significant for Verticillium wilt resistance, pyramiding these elite alleles increased the disease resistance of inbred lines. Furthermore, we selected 34 elite inbred lines, including five lines simultaneously performing elite fiber quality, high yield and resistance to V. dahliae, 14 lines with elite fiber quality and disease resistance, three lines with high yield and disease resistance, and 12 lines with resistance to V. dahliae. No correlation between Verticillium wilt resistance and fiber quality traits/yield and its components was detected in the 155 developed inbred lines. Our results provide candidate markers for disease resistance for use in marker-assisted breeding（MAS）, as well as elite germplasms for improving important agronomic traits via modern cotton breeding.