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.

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.

Genetic variability predicting breeding potential of upland cotton(Gossypium hirsutum L.)for high temperature tolerance

Background High temperature stress at peak flowering stage of cotton is a major hindrance for crop potential.This study aimed to increase genetic divergence regarding heat tolerance in newly developed cultivars and hybrids.Fifty cotton genotypes and 40 F1(hybrids)were tested under field conditions following the treatments,viz.,high temperature stress and control at peak flowering stage in August and October under April and June sowing,respectively.Results The mean squares revealed significant differences among genotypes,treatments,genotype×treatment for relative cell injury,chlorophyll contents,canopy temperature,boll retention and seed cotton yield per plant.The genetic diversity among 50 genotypes was analyzed through cluster analysis and heat susceptibility index(HSI).The heat tolerant genotypes including FH-Noor,NIAB-545,FH-466,FH-Lalazar,FH-458,NIAB-878,IR-NIBGE-8,Weal-AGShahkar,and heat sensitive,i.e.,CIM-602,Silky-3,FH-326,SLH-12 and FH-442 were hybridized in line×tester fashion to produce F1 populations.The breeding materials’populations(40 F1)revealed higher specific combining ability variances along with dominance variances,decided the non-additive type gene action for all the traits.The best general combining ability effects for most of the traits were displayed by the lines,i.e.,FH-Lalazar,NIAB-878 along with testers FH-326 and Silky-3.Specific combining ability effects and better-parent heterosis were showed by the crosses,viz.,FH-Lalazar×Silky-3,FH-Lalazar×FH-326,NIAB-878×Silky-3,and NIAB-878×FH-326 for seed cotton yield and yield contributing traits under high temperature stress.Conclusion Heterosis breeding should be carried out in the presence of non-additive type gene action for all the studied traits.The best combiner parents with better-parent heterosis may be used in crossing program to develop high yielding cultivars,and hybrids for high temperature stress tolerance.

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.

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.

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.

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.

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.

Comparative performance of hybrid generations reveals the potential application of F_(2)hybrids in upland cotton

Background:The utilization of heterosis has greatly improved the productivity of cotton worldwide.However,a major constraint for the large-scale promotion of F_(1) hybrid cotton is artificial emasculation and pollination.This study proposed the potential utilization of F_(2) hybrids to improve upland cotton production through a comparative evaluation of hybrid generations.Results:Eight upland cotton varieties were analyzed and crosses were made according to NCII incomplete diallel cross-breeding design in two cotton belts of China.Variance analysis revealed significant differences in agronomic,yield,and fiber quality in both generations and environments.The broad-sense heritability of agronomic and yield traits was relatively higher than quality traits.Furthermore,the narrow-sense heritability of some traits was higher in F_(2) than in the F_(1) generation in both cotton belts.Overall,parental lines Zhong901,ZB,L28,and Z98 were observed with maximum combining ability while combinations with strong special combining ability were ZB×DT,L28×Z98,and ZB×851.The yield traits heterosis was predominant in both generations.However,the level of heterosis was altered with trait,hybrid combination,generation,and environment.Interestingly,L28×Z98 performed outstandingly in Anyang.Its lint yield(LY)was 24.2%higher in F_(1) and 11.6%in F_(2) than that of the control Ruiza 816.The performance of SJ48×Z98 was excellent in Aral which showed 36.5%higher LY in F_(2)and 10.9%in F_(2)than control CCRI 49.Further results revealed most hybrid combinations had shown a low level of heterosis for agronomic and fiber quality traits in both generations.Comparatively,ZB×DT and L28×Z98 showed hybrid vigor for multiple traits in both generations and cotton belts.It is feasible to screen strong heterosis hybrid combinations with fine fiber in early generations.In the two environments,the correlation of some traits showed the same trend,and the correlation degree of Anyang site was higher than that of Aral site,and the correlation of some traits showed the opposite trend.According to the performance of strong heterosis hybrid combinations in different environments,the plant type,yield and fiber traits associated with them can be improved according to the correlation.Conclusions:Through comparative analysis of variance,combining ability,and heterosis in F_(2)and F_(2)hybrids in different cotton belts,this study proposed the potential utilization of F_(2)hybrids to improve upland cotton productivity in China.

A suppressed gene in integument cells of a fiberless seed mutant in upland cotton

A fiberless seed mutant (fl) was identified in a commercial cotton ( Gossypium hirsutum L.) variety Xu Zhou 142 (FL). This phenotype is associated with lack of fiber cell initiation in the outer integument of the ovule, as was characterized by analysis of genes related to fiber differentiation and development. Two genes, fl E6 and FL E6, were cloned from fl integument cells and FL fiber or integument cells, respectively. Compared with FL E6, fl E6 showed a dramatic change in nucleotide sequence: (1) FL E6 contained a tandem repetitive sequence in which GGCTCA (Gly Ser) is repeated five times between the 82nd and the 93rd codon from the first ATG codon, while in fl E6 the same sequence is repeated four times; (2) The fl E6 gene encodes a polypeptide of 241 amino acids but lacks two codons between the 90th and 93rd codon and three between the 171st and 174th relative to FL E6; (3) There are also 12 nucleotide substitutions which would result in 7 amino acid differences between fl E6 and FL E6. Analysis of RT PCR and Northern Blot showed that expression of the fl E6 gene is suppressed in the fl integument cells, but highly expressed in FL fiber cells. The difference between fl E6 and FL E6 may be associated with lower expression of fl E6 in the fl integument cells. Searches of protein databases with the FL E6 gene sequence showed similarity to the protein backbones of two arabinogalactan proteins (AGPs), one from the filtrate of suspension cultured cells of Pyrus communis (AGPPc2) and the other from Nicotiana alata (AGPNa2). Although the function of the FL E6 protein in differentiation and development of cotton fiber cells is not known, the data indicate that the mutation of fl E6 gene from FL E6 gene may inhibit the fiber cell initiation from epidermal cells of the outer integument of the ovule.

Effect of Implantation Machine Parameters on N^+ ion Implantation for Upland Cotton(Gossypium hirsutum L.)Pollen

Effect of parameters of ion implantation machine, including ion energy, total dose, dose rate, impulse energy and implantation interval on the pollen grains of upland cotton implanted with nitrogen ion beam were studied. The best parameters were screened out. The results also showed that the vacuum condition before the nitrogen ion implantation does not affect the pollen viability.

Pyramiding Breeding by Marker-Assisted Recurrent Selection in Upland Cotton: Selected Effects on Resistance to Helicoverpa armigera

The coincidence rates were more than 96% among the instar-weighted average of bioassaysin the lab, the percentage of resistance to Km in the field and the percentage of plantscontaining Bt gene. So, the performance of resistance to Km in the field can be used torepresent the transgenic Bt gene for selecting the resistance to bollworm. The instar-weighted averages were 30.585, 24.182, 16.615, 10.601, 10.123, 7.440 and 7.215 for theC0, P1, M1, M2, MP1, P2 and MP2 populations, respectively. The variance analysisindicated that the instar-weighted average in C0 was greatly significantly higher thanthat in all other populations, i.e., the performance of resistance to bollworm in C0 washighly significantly lower than all other populations. And the resistance in P1 wasgreatly lower than that of M1, M2, MP1, P2 and MP2, and M1 greatly lower than that of M2,MP1, P2 and MP2. There were no significant differences among M2, MP1, P2 and MP2. Withinthe populations of the first cycle selection, MP1 and M1 were greatly significantlyhigher than P1, and MP1 significantly higher than M1. The populations of the second cycleselection were significantly higher than their initial population M1, but no significantdifference among them. The boll size, seed index, the percent of the first harvest yield,fiber length, strength and elongation of the resistant plants to bollworm were significantlylower than that of sensitive plants to bollworm. And the yield of seed and lint cottonof the resistant plant to bollworm were lower than that of the sensitive to bollworm, butno significant difference between them. The boll numbers per plant, lint percent andmicronaire of the resistant plants to bollworm were significantly higher than that of thesensitive plant to bollworm.

Development of Transgenic Restorer of Cytoplasmic Male Sterility in Upland Cotton

A glutathione S-transferase gene (gst) has been introduced into restorers of cytoplasmic male sterility in upland cotton (Gossypium hirsutum L.) using Agrobacterium-mediated transformation. A trans-genie restorer, signed as 'Zheda strong restorer', which has strong restorability to male sterility, was selected from progeny plants of transformants. When compared with an American restorer 'DES-HAF277', the fertility restorability of 'Zheda strong restorer' to male sterility was been enhanced by 25.8% in the percentage of viable pollens of hybrid (sterile line × restorer) F1 The hybrid gave 3.6 more bolls per plant, 10.1% less aborted seeds and 10.6% more lint yield when 'Zheda strong restorer' was used as male parent than when 'DES-HAF277' was. Southern and Northern bloting analysis showed that the foreign gsf gene was detectable and highly expressed in 'Zheda strong restorer'.

QTL and genetic analysis controlling fiber quality traits using paternal backcross population in upland cotton

Background:Genetic improvement in fiber quality is one of the main challenges for cotton breeders.Quantitative trait loci(QTL)mapping provides a powerful approach to dissect the molecular mechanism in fiber quality traits.In present study,F14 recombinant inbred line(RIL)population was backcrossed to paternal parent for a paternal backcross(BC/P)population,deriving from one upland cotton hybrid.Three repetitive BC/P field trials and one maternal backcross(BC/M)field trial were performed including both two BC populations and the original RIL population.Results:In total,24 novel QTLs are detected for fiber quality traits and among which 13 QTLs validated previous results.Thirty-five QTLs in BC/P populations explain 5.01%–22.09%of phenotype variation(PV).Among the 35 QTLs,23 QTLs are detected in BC/P population alone.Present study provides novel alleles of male parent for fiber quality traits with positive genetic effects.Particularly,qFS-Chr3–1 explains 22.09%of PV in BC/P population,which increaseds 0.48 cN·tex−1 for fiber strength.A total of 7,2,8,2 and 6 QTLs explain over 10.00%of PV for fiber length,fiber uniformity,fiber strength,fiber elongation and fiber micronaire,respectively.In RIL population,six common QTLs are detected in more than one environment:qFL-Chr1–2,qFS-Chr5–1,qFS-Chr9–1,qFS-Chr21–1,qFM-Chr9–1 and qFM-Chr9–2.Two common QTLs of qFE-Chr2–2(TMB2386-SWU12343)and qFM-Chr9–1(NAU2873-CGR6771)explain 22.42%and 21.91%of PV.The region between NAU4034 and TMB1296 harbor 30 genes(379 kb)in A05 and 42 genes(49 kb)in D05 for fiber length along the QTL qFL-Chr5–1 in BC/P population,respectively.In addition,a total of 142 and 46 epistatic QTLs and QTL×environments(E-QTLs and QQEs)are identified in recombinant inbred lines in paternal backcross(RIL-P)and paternal backcross(BC/P)populations,respectively.Conclusions:The present studies provide informative basis for improving cotton fiber quality in different populations.

Intron-Targeted Intron-Exon Splice Conjunction (IT-ISJ) Marker and Its Application in Construction of Upland Cotton Linkage Map

To develop a new DNA maker, which could be used in genetic diversity analysis and genetic map construction in plants, IT-ISJ （intron targeted intron-exon splice junction） primer combinations, which were designed according to the intronexon splice junction conserved sequences, were used to construct cotton genetic linkage map in the present study. 49 out of 704 IT-ISJ primer combinations showed polymorphism between upland cotton high quality cultivar Yumian 1 and multiple dominant gene line T586, and the polymorphic primer combinations accounted for 7.0% of total primer combinations. 49 IT-ISJ primer combinations were used to genotype 270 F2：7 recombinant inbred lines developed from （Yumian 1 × T586） F2, and 58 IT-ISJ loci were obtained. 58 IT-ISJ, together with 150 SSR and 8 morphological loci, were used to conduct linkage analysis, and a linkage map including 22 linkage groups and 113 loci （49 IT-ISJ, 62 SSR, and 2 morphological loci） was constructed. The linkage map covered 714.5 cM with an average interval of 6.3 cM between two markers, accounting for 16.1% of cotton genome. The present study demonstrated that the polymorphism of IT-ISJ marker is high, and it could be effectively applied in plant genetic map construction.