Investigation of GhFAT Genes Related to Seed Oil Content and Fatty Acid Composition in Gossypium hirsutum L.

Fatty Acyl-ACP thioesterase(FAT)is a key enzyme controlling oil biosynthesis in plant seeds.FATs can be divided into two subfamilies,FATA and FATB according to their amino acid sequences and substrate specificity.The Upland cotton genome contains 20 GhFAT genes,amongst which 6 genes were of the GhFATA subfamily and 14 of the GhFATB subfamily.The 20 GhFAT genes are unevenly distributed on 14 chromosomes.The GhFATA genes have 5 or 7 exons and the GhFATB genes have 6 or 7 exons.All GhFAT proteins have the conserved Acyl-ACP_TE domain and PLN02370 super family,the typical characteristics of plant thioesterases.Analyses of the expression level of GhFATs and the compositions of fatty acid in 5-60 days-post-anthesis seeds showed that the ratio of saturated fatty acids to unsaturated fatty acids was consistent with the expression profile of GhFATB12,GhFATB3,and GhFATB10;the ratio of monounsaturated fatty acid to polyunsaturated fatty acids was consistent with the expression profile of GhFATA3.The oil contents of mature cottonseeds were positively correlated with the contents of palmitic acid and linolenic acid as well as seed vigor.These results provide essential information for further exploring the role(s)of the specific GhFATs in determining oil biosynthesis and cottonseed compositions.

多环境下陆地棉(Gossypium hirsutum L.)重组自交系铃重与衣分性状的QTL分析

本研究利用以SGK9708为母本,0-153为父本构建的196个陆地棉重组自交系(F6:8)构建了包含186个标记,总长827.84cM,标记间平均距离4.45cM,覆盖棉花基因组18.6%的遗传连锁图谱,并对7个环境下的铃重和衣分性状进行QTL定位和上位性互作分析。利用两种分析软件(WinQTLcart2.5和QTLNetwork2.0)共同定位了多个环境下稳定表达的5个主效QTLs(qBW-1-1,qBW-1-2,qLP-2-1,qLP-2-2和qLP-4-2)。利用QTLNetwork2.0分别检测到4对铃重上位性互作QTLs和7对衣分上位性互作QTLs,以背景位点间的互作和加性效应位点与背景位点间的互作为主。除主效QTL外,上位性效应也是陆地棉铃重和衣分性状的重要遗传基础。本研究定位的5个主效QTLs为选择高铃重、高衣分品种的棉花分子标记辅助育种提供了重要依据。

棉花(Gossypium hirsutum L.)原生质体培养的体细胞胚胎发生及植株再生

以陆地棉(G.hirsutum L.)3118、晋棉4号和柯字棉312等3个品种为材料,取种子无菌苗的下胚轴诱导愈伤组织,建立胚性细胞悬浮培养系。以2％纤维素酶Ono-zuka RS和0.1％离析软化酶Y-23组成的混合酶液,从细胞悬浮培养物游离出原生质体。用含低融点琼脂糖的K_3培养基包埋原生质体,采用园柱及薄层漂浮平板培养方式。3个品种的原生质体培养都得到了愈伤组织,其中晋棉4号和柯字棉312品种的愈伤组织转到低激素水平或无激素的MS-2培养基上诱导产生胚状体,胚状体在无激素的MS-1培养基上形成了植株。

Screening of drought resistance indices and evaluation of drought resistance in cotton(Gossypium hirsutum L.)

Sixteen cotton cultivars widely planted in China were sowed under five different drought concentrations(0,2.5,5,7.5,and 10%)using PEG6000 to screen the indices of drought resistance identification and explore the drought resistance of different cotton cultivars.Eighteen physiological indices including root,stem,and leaf water contents(RWC,SWC,and LWC),net photosynthetic rate(Pn),the maximum photochemical quantum yield(Fv/Fm),the actual photochemical quantum yield(ΦPSII),non-photochemical quenching coefficient(NPQ),leaf water potential(LWP),osmotic potential(Ψs),leaf relative conductivity(REC),leaf proline content(Pro),leaf and root soluble protein contents(LSPC and RSPC),leaf and root malondialdehyde(MDA)contents(LMDA and RMDA),root superoxide dismutase,peroxidase,and catalase activities(RSOD,RPOD,and RCAT)were measured.Results indicated the 18 physiological indices can be converted into five or six independent comprehensive indices by principal component analysis,and nine typical indices(Fv/Fm,SWC,LWP,Pro,LMDA,RSPC,RMDA,RSOD,and RCAT)screened out by a stepwise regression method could be utilized to evaluate the drought resistance.Moreover,the 16 cotton cultivars were divided into four types:drought sensitive,drought weak sensitive,moderate drought resistant,and drought resistant types.The resistance ability of two selected cotton cultivars(drought resistant cultivar,Dexiamian 1;drought sensitive cultivar,Yuzaomian 9110)with contrasting drought sensitivities were further verified by pot experiment.Results showed that the responses of final cotton biomass,yield,and yield composition to drought were significantly different between the two cultivars.In conclusion,drought resistant cultivar Dexiamian 1 and drought sensitive cultivar Yuzaomian 9110 were screened through hydroponics experiment,which can be used as ideal experimental materials to study the mechanism of different cotton cultivars with contrasting drought sensitivities in response to drought stress.

棉花(Gossypium hirsutum L.)花粉壁的发育

用光镜和透射电镜观察研究了棉花花粉壁的发育过程.棉花花粉壁的发生始于四分孢子时期,四分体形成后,在小孢子的质膜与胼胝质壁之间逐渐形成多糖性质的原外壁.游离小孢子时期,在一定间隔的原外壁中小孢子质膜表面积累外壁物质并形成基粒棒,之后在基粒棒的下部和上部分别横向延展并接合形成基足层和覆盖层.花粉内壁发生时期较晚,大约在大液泡形成之前.但内层和外层花粉壁均在小孢子有丝分裂之前同时建成.构建花粉壁的物质主要来自绒毡层的周原质团和小孢子原生质体的代谢分泌.小孢子胞质中在四分体时期形成的巨型内质网复合体是较为特殊的细胞结构,在游离小孢子时期,巨型内质网不断产生大量的内含微粒物的运输小泡,这种小囊泡表现出向质膜趋向运动并进一步与质膜融合.推测由内质网复合体产生的这些小囊泡是小孢子原生质体向质膜外运输花粉壁构建物质的主要形式.

Relationship between plant canopy characteristics and photosynthetic productivity in diverse cultivars of cotton(Gossypium hirsutum L.)

Genotype and plant type affect photosynthetic production by changing the canopy structure in crops.To analyze the mechanism of action of heterosis and plant type on canopy structure in cotton(Gossypium hirsutum L.),we had selected two cotton hybrids(Shiza 2,Xinluzao 43) and two conventional varieties(Xinluzao 13,Xinluzao 33) with different plant types in this experiment.We studied canopy characteristics and their correlation with photosynthesis in populations of different genotypes and plant types during yield formation in Xinjiang,China.Canopy characteristics including leaf area index(LAI),mean foliage tilt angle(MTA),canopy openness(DIFN),and chlorophyll relative content(SPAD).The results showed that LAI and SPAD peak values were higher and their peak values arrived later,and the adjustment capacity of MTA during the flowering and boll-forming stages was stronger in Xinluzao 43,with the normal-leaf,pagoda plant type,than these values in other varieties.DIFN of Xinluzao 43 remained between0.09 and 0.12 during the flowering and boll-forming stages,but was lower than that in the other varieties during the boll-opening stage.Thus,these characteristics of Xinluzao 43 were helpful for optimizing the light environment and maximizing light interception,thereby increasing photosynthetic capability.The photosynthetic rate and photosynthetic area were thus affected by cotton genotype as changes in the adjustment range of MTA,increases in peak values of LAI and SPAD,and extension of the functional stage of leaves.Available photosynthetic area and canopy light environment were affected by cotton plant type as changes in MTA and DIFN.Heterosis expression and plant type development were coordinated during different growth stages,the key to optimizing the canopy structure and further increasing yield.

Somatic Embryogenesis and Plant Regeneration from Two Recalcitrant Genotypes of Gossypium hirsutum L.

An improved protocol has been developed for somatic embryogenesis and plant regeneration of recalcitrant cotton cultivars. High callus frequencies and embryogenic tissue were developed in MSB medium supplemented with gradient concentrations of KT and 2,4-D, their concentration decreasing from 0.1 to 0.01 mg·L^-1. Somatic embryos were successfully incubated in 1/2 macronutrient MSB suspension supplemented with 0.5 g· L^-1 glutamine and 0.5 g·L^-1 asparagine. Decrease in macronutrient concentration of MSB significantly alleviated browning and was beneficial to suspension cells. Transformation of somatic embryos into plants was induced in MSB medium supplemented with 3% sucrose, 0.5 g·L^-1 glutamine, 0.5 g·L^-1 asparagine, and 6.0 g·L^-1 agar. The effect of sucrose as carbohydrate was better than that of glucose for plant germination. Using this protocol, regenerated plantlets from the CCRI521 and Zhongzhi86-6 reached to as much as 19.6 and 18.5% somatic embryos, respectively.

Molecular Cloning and Characterization of Three Novel Genes Related to Fatty Acid Degradation and Their Responses to Abiotic Stresses in Gossypium hirsutum L.

Fatty acid metabolism is responsible not only for oilseed metabolism but also for plant responses to abiotic stresses. In this study, three novel genes related to fatty acid degradation designated GhACX, Gh4CL, and GhMFP, respectively, were isolated from Gossypium hirsutum acc. TM-1. The phylogenetic analysis revealed that amino acid sequences of GhACXand GhMFP have the highest homology with those from Vitis vinifera, and Gh4CL has a closer genetic relationship with that from Camellia sinensis. Tissue- and organ-specific analysis showed that the three genes expressed widely in all the tested tissues, including ovules and fiber at different developing stages, with expressed preferentially in some organs. Among them, GhACX showed the most abundant transcripts in seeds at 25 d post anthesis （DPA）, however, GhMFP and Gh4CL have the strongest expression level in ovules on the day of anthesis. Based on real-time quantitative RT-PCR, the three genes were differentially regulated when induced under wounding, methyl jasmonate （MeJA）, cold, and abscisic acid （ABA） treatments. The characterization and expression pattern of three novel fatty acid degradation related genes will aid both to understand the roles of fatty acid degradation related genes as precursor in stress stimuli and to elucidate the physiological function in cotton oilseed metabolism.

Introduction of Bacillus thuringiensis(Bt)gene does not reduce potassium use efficiency of Bt transgenic cotton(Gossypium hirsutum L.)

Background:Potassium(K)deficiency has become a common field production problem following the widespread adoption of Bacillus thuringiensis(Bt)transgenic cotton(Gossypium hirsutum L.)worldwide.The purpose of this study was to clarify whether the introduction of Bt gene directly reduces the K-use efficiency of cotton to induce K deficiency.Results:The cotton variety,Jihe 321(wild type,WT)and its two Bt(Cry1Ac)-transgenic overexpression lines(OE-29317,OE-29312)were studied in field with low soil-test K+(47.8 mg·kg^(−1)).In the field with low soil-test K+,only OE-29317 had less biomass and K+accumulation than the WT at some growth stages.Both Bt lines produced similar or even greater seed cotton yield than WT in the field.When the Bt gene(~70%)in OE-29317 and OE-29312 plants was silenced by virus-induced gene silencing(VIGS),the VIGS-Bt plants did not produce more biomass than VIGSgreen fluorescent protein(control)plants.Conclusions:The introduction of Bt gene did not necessarily hinder the K use efficiency of the cotton lines under this study.

陆地棉(Gossypium hirsutum L.)单体的诱发和鉴定

以隐性亲本(芽黄 v_1,v_6,v_8,v_(10),v_(11),T582,TM-1)作母本,授以辐射处理的显性亲本(TM-1,2256-61,陕1155,泗棉2号,Rd,T586)的花粉,进行单体的诱发研究。通过对 F_1表型为隐性或畸形的植株,在现蕾时观察其减数分裂时的染色体构型,发现有单倍体的存在。并将单体作母本,用相应的易位系测验,鉴定了诱发得到的单体株分别为单体1、6、7。辐射剂量为800伦琴和500伦琴,其诱致单体的频率分列为20.5%和9.8%。发现高剂量比低剂量更能诱致染色体的丢失,特别是小染色体的丢失,并且高剂量诱致的染色体结构变异也增加。

Genome-wide identification and expression analysis of the GhIQD gene family in upland cotton(Gossypium hirsutum L.)

Background:Calmodulin(CaM)is one of the most important Ca^(2+)signaling receptors because it regulates diverse physiological and biochemical reactions in plants.CaM functions by interacting with CaM-binding proteins(CaMBPs)to modulate Ca^(2+)signaling.IQ domain(IQD)proteins are plant-specific CaMBPs that bind to CaM by their specific CaM binding sites.Results:In this study,we identified 102 GhIQD genes in the Gossypium hirsutum L.genome.The GhIQD gene family was classified into four clusters(Ⅰ,Ⅱ,Ⅲ,andⅣ),and we then mapped the GhIQD genes to the G.hirsutum L.chromosomes.Moreover,we found that 100 of the 102 GhIQD genes resulted from segmental duplication events,indicating that segmental duplication is the main force driving GhIQD gene expansion.Gene expression pattern analysis showed that a total of 89 GhIQD genes expressed in the elongation stage and second cell wall biosynthesis stage of the fiber cells,suggesting that GhIQD genes may contribute to fiber cell development in cotton.In addition,we found that 20 selected GhIQD genes were highly expressed in various tissues.Exogenous application of MeJA significantly enhanced the expression levels of GhIQD genes.Conclusions:Our study shows that GhIQD genes are involved in fiber cell development in cotton and are also widely induced by MeJA.Thw results provide bases to systematically characterize the evolution and biological functions of GhIQD genes,as well as clues to breed better cotton varieties in the future.

Genome-wide association study of micronaire using a natural population of representative upland cotton(Gossypium hirsutum L.)

Background: Micronaire is a comprehensive index reflecting the fineness and maturity of cotton fiber.Micronaire is one of the important internal quality indicators of the cotton fiber and is closely related to the value of the cotton fiber.Understanding the genetic basis of micronaire is required for the genetic improvement of the trait.However,the genetic architecture of micronaire at the genomic level is unclear.The present genome-wide association study(GWAS)aimed to identify the genetic mechanism of the micronaire trait in 83 representa:tive upland cotton lines grown in multiple environments.Results GWAS of micronaire used 83 upland cotton accessions assayed by a Cotton 63 K Illumina Infinium single nucleotide polymorphism(SNP)array.A total of 11 quantitative trait loci(QTLs)for micronaire were detected on 10 chromosomes.These 11 QTLs included 27 identified genes with specific expression patterns.A novel QTL,qFM-A12–1,included 12 significant SNPs,and GhFLA9 was identified as a candidate gene based on haplotype block analysis and on strong and direct linkage disequilibrium between the significantly related SNPs and gene.GhFLA9 was expressed at a high level during secondary wall thickening at 20∼25 days post-anthesis.The expression level of GhFLA9 was significantly higher in the low micronaire line(Msco-12)than that in the high micronaire line(Chuangyou-9).Conclusions: This study provides a genetic reference for genetic improvement of cotton fiber micronaire and a foundation for verification of the functions of GhFLA9.

Evaluation of genetic diversity in short duration cotton(Gossypium hirsutum L.)

Background: Cotton(Gossypium hirsutum L.) is an important fiber crop in Bangladesh. Genetic diversity among the genotypes of a germplasm has a great importance for cotton breeding. An experiment was carried out at the experimental field of Cotton Research, Training and Seed Multiplication Farm, Sreepur, Gazipur during the cropping season of 2015-2016 with 100 genotypes to evaluate genetic diversity of cotton genotypes for short duration using field performance.Results: The genotypes under study were grouped into ten clusters through multivariate analysis using GENSTAT-5.Cluster III contained maximum number of genotypes(16) while cluster X contained the least number of genotypes(7).The inter cluster distances were larger than intra cluster distances in all cases suggesting wider genetic diversity among the genotypes of different clusters. The maximum and minimum inter cluster distances were observed between clusters II and V(10.78) and clusters VIII and IX(3.30), respectively. The results indicated diverse and close relationship among the genotypes of those clusters. Earliness index, single boll weight and days to boll opening showed the higher contribution to the genetic divergence among 19 characters.Conclusion: Based on the results of genetic diversity and earliness index, the genotypes from cluster Ⅱ could be used as parent in hybridization program for the development of short duration cotton variety.

Ameliorative effects of potassium on drought-induced decreases in fiber length of cotton(Gossypium hirsutum L.) are associated with osmolyte dynamics during fiber development

Fiber length of cotton(Gossypium hirsutum L.)decreases under drought stress,potassium(K)could diminish the decreased caused by drought,but the mechanism associated with this alleviation effect is not clear.We evaluated the effect of K on fiber elongation using two cotton cultivars,Simian 3 and Siza 3,grown in well-watered and drought-stressed conditions.Potassium fertilizer(K2O)was applied 0,150,or 300 kg ha?1 in each growing condition.Drought stress reduced the final fiber length due to a decline in the maximum rate of rapid elongation(Vmax,mmday?1).The application of K alleviated the droughtinduced fiber length reduction by increasing Vmax.At 10 and 15 days post-anthesis(DPA),drought significantly reduced osmotic potential(OP)and increased K+and malate contents at all K rates,relative to well-watered conditions,which was associated with increased activities of phosphoenolpyruvate carboxylase(PEPC),V-ATPase,PPase,and PM H+-ATPase in cotton fiber.However,the relative contribution of K+and malate to OP declined under drought in comparison with well-watered condition.Compared with control without K,K application decreased OP and increased the accumulation of osmolytes(K+,malate and soluble sugar)as well as the activities of related enzymes in fiber irrespective of water treatments.Moreover,K application increased osmotic adjustment during drought,and improved the contribution of K+and malate to OP,especially under drought stress.This study showed that drought decreased fiber length by reducing Vmax,and K application ameliorates the decline in fiber elongation due to drought by enhancing osmolytes accumulation and their contribution to OP in fiber cells.

Comparative Analysis of Salinity-Induced Proteomic Changes in Cotton (Gossypium hirsutum L.)

Salt stress on cotton varieties of distinct salinity tolerance can induce expression of different proteins. Zhong 07, a salt-tolerant variety and Zhong s9612, a salt-sensitive variety, were utilized as experimental materials. The leaves of trefoil seedlings treated with or without 0.4% NaCl for 24 h were harvested for whole-protein extraction. Two-dimensional technology, combined with mass spectroscopy (MS) analysis and protein database searching, was employed to detect differentially expressed proteins and determine their identities and biological functions. Compared with the control, Zhong 07 showed 10 differentially expressed proteins under salt stress, of which 6 were upregulated and 4 were downregulated. Meanwhile, 12 differentially expressed proteins were detected in Zhong s9612 under salt stress, of which 10 were upregulated and 2 were downregulated. In the matrix-assisted laser desorption-ionization/time of flight-time of flight/MS analysis, 14 differentially expressed proteins were successfully identified, including the ribulose-1, 5-bisphosphate carboxylase/oxygenase (RuBisco) large subunit-binding protein subunit alpha (RuBisco α), luminal binding protein (LBP), heat shock protein 70 (Hsp1, 2, 3), pathogenesis-related protein class 10 (PR-10), quinoneoxidoreductase-like protein (QOR), S-adenosylmethioninesyn-thetase (SAMS), enolase (EN), and RuBisco large subunit-binding protein subunit beta (RuBisco β). Cellular function is ultimately executed by proteins, and cotton varieties with different salt tolerance can be influenced by salt stress to various degrees, which can provide certain theoretical foundation for the identification of salt tolerance of cotton varieties. The findings also provide some proteins, such as the RuBisco large subunit binding proteins α and β subunits, OEE2 protein, HSP70, and S-adenosylmethionine synthetase, which can be used as protein markers of salt-to-lerance before- and post-treatment, making a big difference in salt-tolerance identification in cotton.

An Integrated Genetic,Physical and Transcript Map of Homoeologous Chromosomes 12 and 26 in Upland Cotton(Gossypium hirsutum L.)

While Upland cotton(Gossypium hirsutum L.) represents 95% of the world production,its genetic improvement is hindered by the shortage of effective genomic tools and resources.The

Selection and Characterization of a Novel Glyphosate Tolerant Upland Cotton(Gossypium hirsutum L.) Mutant(R1098)

Stepwise selection approach was adopted to obtain glyphosate-tolerant upland cotton mutant(R1098) from the embryogenic calli of Coker 312(Gossypium hirsutum L.).The calli were transferred to selection medium and multi-step selection pressure process was carried out until the

Biochemical Genetic Mechanism and QTLs of Early Maturing without Yield Loss in Short-season Upland Cotton(Gossypium hirsutum L.)

The short season cotton(SSC) was important Upland plant ecotype(Gossypium hirsutum L.).The growth of SSC was very short that is 105 ～ 110 days(after planting). SSC could increase

Cloning and Function Characteristic of GhDWF4,an Ortholog of Arabidopsis DWF4 from Upland Cotton(Gossypium hirsutum L.)

As one of the longest cells characterized in plant kingdom,cotton fibers were regarded as an ideal material for studying plant cell growth and development.In recent years,several reports revealed

Heat Stress Related Gene Expression in Gossypium hirsutum L.

Abiotic stress is a major limiting factor to crop productivity,and heat stress is one of the important elements for reduced crop production.Plants respond to heat stress at molecular and cellular levels as well as physiological level.Heat stress alters expression patterns of numerous genes in plants.