陆地棉EST-SSRs在向日葵中的通用性研究

利用陆地棉EST-SSR s(Expressed sequence tags-s im p le sequence repeats)在陆地棉和向日葵品种中进行了通用性分析。利用本实验室合成的1 500对陆地棉EST-SSR s在4个陆地棉品种中进行了扩增,有效扩增比率为95%,扩增产物符合预期大小(50~500 bp);每对引物可以检测到1~6个数目不等的片段(a lle le),平均约为2.7个;每2个陆地棉之间大约有7%~9%的多态性。从1 500对陆地棉EST-SSR s引物中选取400对引物在向日葵中进行了扩增,有效扩增比率为63%,每对引物可以检测到1~6个数目不等的片段,平均为1.7个,片段大小介于50~500 bp之间。选取了200对在向日葵中得到有效扩增的引物对向日葵G 101的父母本进行了多态性筛选,约25%引物具有多态性。结果表明,陆地棉EST-SSR s在向日葵中具有较高的通用性,可用于向日葵比较基因组研究和分子标记研究。     

Genomic divergence in cotton germplasm related to maturity and heterosis

Commercial varieties of upland cotton(Gossypium hirsutum) have undergone extensive breeding for agronomic traits, such as fiber quality, disease resistance,and yield. Cotton breeding programs have widely used Chinese upland cotton source germplasm(CUCSG) with excellent agronomic traits. A better understanding of the genetic diversity and genomic characteristics of these accessions could accelerate the identification of desirable alleles. Here, we analyzed 10,522 high-quality singlenucleotide polymorphisms(SNP) with the CottonSNP63 K microarray in 137 cotton accessions(including 12 hybrids of upland cotton). These data were used to investigate the genetic diversity, population structure,and genomic characteristics of each population and the contribution of these loci to heterosis. Three subgroups were identified, in agreement with their knownpedigrees, geographical distributions, and times since introduction. For each group, we identified lineagespecific genomic divergence regions, which potentially harbor key alleles that determine the characteristics of each group, such as early maturity-related loci. Investigation of the distribution of heterozygous loci, among 12 commercial cotton hybrids, revealed a potential role for these regions in heterosis. Our study provides insight into the population structure of upland cotton germplasm. Furthermore, the overlap between lineagespecific regions and heterozygous loci, in the high-yield hybrids, suggests a role for these regions in cotton heterosis.     

用RAPD标记评估我国棉花品种遗传多样性

利用ＲＡＰＤ标记对１７个不同来源的我国棉花品种（系）进行遗传多样性评估。２００个随机引物扩增出１１３条多态性片段。１７个品种（系）依据染色体组的不同可以分为３大类。它们之间遗传相似性系数从０．２０００到０．８４５１。３个大面积推广品种鄂荆１号,中棉所１２号和苏棉３号遗传基础十分相近。陆地棉与阔叶棉杂交产生的野生种质系７５８１,与其他陆地棉品种（系）遗传差异显著。不同育种单位在育种过程中种质使用具有     

陆地棉超矮杆突变性状质量遗传规律分析

分析了陆地棉超矮突变体的质量遗传规律, 结果显示, 该性状是受一对完全隐性基因控制的质量性状; 在国内外未见陆地棉中同类突变的报道, 暂将它的基因符号定为du。该突变体在自然条件下生长, 最终株高仅10.5 cm, 不能正常开花、结铃和吐絮; 但通过外源激素处理, 最终株高可以达57.8 cm, 并能正常结铃吐絮。     

建国以来我国棉花品种遗传基础的分子标记分析

采用MPD分子标记、遗传距离和聚类分析方法,研究建国以来我国有代表性的166个棉花主栽品种(或品系)的遗传多样性。41个RAPD标记Nei’s遗传距离(GD)与两组不同来源实验数据的表型性状欧氏距离(UD)间相关系数分别为0．6445(n＝1770)和0．7078(n＝7140),表明RAPD可以揭示棉花品种间遗传差异。通过对不同棉种、不同品种类型、不同时期、不同种植区域和不同来源的棉花品种(系)遗传差异的比较,探讨我国棉花品种的遗传基础。各层次上遗传差异的比较表明：在我国主栽棉花品种中,海岛棉品种遗传基础窄于陆地棉品种;我国自育陆地棉品种的遗传基础窄于国外引进品种;杂交陆地棉品种的遗传基础窄于常规品种;上世纪80年代以后陆地棉品种遗传基础窄于70年代品种;长江棉区品种遗传基础窄于黄淮棉区品种,西北内陆棉区品种窄于长江棉区品种。启示我们如何在我国棉花育种的全局和不同层面上把握和制定拓宽棉花育种遗传基础的策略和手段,并为进步深入探讨建国以来我国棉花品种遗传改良规律打下基础。     

海岛棉与陆地棉叶绿素含量变化的差异研究

以5份陆地棉和4份海岛棉为材料,研究了整个发育进程中陆地棉和海岛棉叶绿素的变化规律之间的差异,结果显示：陆地棉与海岛棉的叶绿素a、叶绿素b和叶绿素a＋b各有不同动态变化规律,其叶绿素a/b在整个发育进程中的变化也有差异。海岛棉与陆地棉的叶绿素a变化趋势有较大差别,陆地棉的叶绿素a含量在6月18日（苗期）最高,峰值出现在8月6日,海岛棉峰值、最大值均在7月4日（现蕾期）;叶绿素b含量表现为陆地棉、海岛棉峰值均在8月6日（花铃期）,但海岛棉平均值高于陆地棉;陆地棉的叶绿素a＋b和叶绿素a曲线相似,海岛棉的叶绿素a＋b和叶绿素b曲线相似;陆地棉叶绿素a/b值表现苗期最大,然后迅速下降,海岛棉a/b值表现普遍低于陆地棉,其变化趋势为前期在7月4日出现峰值,然后下降再升高。实验说明陆地棉和海岛棉叶绿素的合成机制、调控机理可能不同。     

Genome- Wide Analysis and Characterization of the TRX Gene Family in Upland Cotton

Thioredoxins (TRX) are small molecules of proteins that are present in all organisms. TRXs play an important role in diverse functions of plant growth and development. In this study, we performed genome-wide, characterization and expression levels of TRX gene family in cotton. A total of 150 GhTRX proteins were identified in upland cotton and classified into five subfamilies based on their domain compositions. Phylogenetic tree analysis divided TRX genes into seven subgroups. GhTRX genes covered all upland cotton chromosomes, with duplicated gene events. Ka/Ks ratio of three gene pairs was less than 1, suggesting purifying selection. The functions of GhTRX genes were studied using gene ontology, protein localization, and promoter analysis. Furthermore, six GhTRX genes were randomly selected to examine their expression level in cotton development and under various exogenous treatments. The genes showed high expressions in various tissues and at different stages of leaf senescence, also showed high expression under abscisic acid, ethylene, drought, and salinity. This study reveals the first report of TRX family genes in upland cotton. However further studies are needed to elucidate their specific functions in cotton plant.     

转FBP7::iaaM基因陆地棉种质冀资139纤维品质性状杂种优势分析

提高棉花(Gossypium hirsutum)产量兼顾改良纤维品质是棉花育种的重要目标, 而优良种质创新是品种改良的基础。FBP7::iaaM基因能够调控棉花胚珠表皮细胞IAA的含量, 进而促进棉纤维发育的起始。利用含有FBP7::iaaM基因的种质IF1-1, 通过常规杂交育种手段实现了目的基因向骨干亲本的转移, 培育了优良陆地棉种质冀资139, 并用4个不同类型的陆地棉品系对冀资139纤维品质性状进行了杂种优势分析。结果表明, FBP7::iaaM基因及其调控的优良性状可以在骨干育种亲本中传递, 具有较高的育种价值; 转FBP7::iaaM基因的冀资139具有综合性状优良、优质和高衣分等特点; 纤维长度、比强度及马克隆值的遗传主要由基因加性效应控制, 这为其作为骨干亲本的应用提供了理论依据。     

Genomic analyses reveal the genetic basis of early maturity and identification of loci and candidate genes in upland cotton (Gossypium hirsutum L.)

Although upland cotton (Gossypium hirsutism L.) originated in the tropics, this early maturity cotton can be planted as far north as 46°N in China due to the accumulation of numerous phenotypic and physiological adaptations during domestication. However, how the genome of early maturity cotton has been altered by strong human selection remains largely unknown. Herein, we report a cotton genome variation map generated by the resequencing of 436 cotton accessions. Whole‐genome scans for sweep regions identified 357 putative selection sweeps covering 4.94% (112 Mb) of the upland cotton genome, including 5184 genes. These genes were functionally related to flowering time control, hormone catabolism, ageing and defence response adaptations to environmental changes. A genome‐wide association study (GWAS) for seven early maturity traits identified 307 significant loci, 22.48% (69) of which overlapped with putative selection sweeps that occurred during the artificial selection of early maturity cotton. Several previously undescribed candidate genes associated with early maturity were identified by GWAS. This study provides insights into the genetic basis of early maturity in upland cotton as well as breeding resources for cotton improvement.     

田间条件下海岛棉和陆地棉花铃期叶片光保护的机制

研究海岛棉(Gossypium barbadense)和陆地棉(G. hirsutum)两个棉花栽培种的光合作用特性, 探讨两个栽培种光合机构的光抑制以及防御保护机制, 以期为新疆棉花高光效品种选育和高产高效栽培实践提供理论基础。在新疆生态气候条件下, 系统测定了海岛棉和陆地棉的叶片运动、叶片接受光量子通量密度(PFD)、叶片温度、叶绿素荧光参数、气体交换参数和光呼吸速率的日变化。研究结果表明: 陆地棉叶片的“横向日性”较强而海岛棉较弱, 导致海岛棉叶片接受PFD较低, 但其叶片温度较高。海岛棉叶片的光合速率和气孔导度均显著低于陆地棉。在8:00-10:00 (北京时间, 下同)海岛棉叶片的光呼吸速率略低于陆地棉, 其余时间段海岛棉和陆地棉叶片的光呼吸速率相似。不同栽培种间, 叶片的最大光化学效率和实际光化学效率的日变化均无明显差异。除14:00-16:00以外, 海岛棉叶片的表观电子传递速率和光化学猝灭系数均显著低于陆地棉。8:00以后, 海岛棉叶片的非光化学猝灭显著高于陆地棉。因此, 在新疆生态气候条件下, 海岛棉和陆地棉叶片“横向日性”运动能力和气孔导度的差异导致叶片所处的光温环境不同, 同时造成海岛棉叶片的碳同化能力较低。为阻止光合电子传递链的过度还原, 减轻光合机构的光抑制, 陆地棉叶片主要通过光合机构的电子流途径耗散激发能, 而海岛棉叶片通过热耗散途径和相对较高的光呼吸能力来耗散激发能。     

Island Cotton Gbve1 Gene Encoding A Receptor-Like Protein Confers Resistance to Both Defoliating and Non-Defoliating Isolates of Verticillium dahliae

Verticillium wilt caused by soilborne fungus Verticillium dahliae could significantly reduce cotton yield. Here, we cloned a tomato Ve homologous gene, Gbve1, from an island cotton cultivar that is resistant to Verticillium wilt. We found that the Gbve1 gene was induced by V. dahliae and by phytohormones salicylic acid, jasmonic acid, and ethylene, but not by abscisic acid. The induction of Gbve1 in resistant cotton was quicker and stronger than in Verticillium-susceptible upland cotton following V. dahliae inoculation. Gbve1 promoter-driving GUS activity was found exclusively in the vascular bundles of roots and stems of transgenic Arabidopsis. Virus-induced silencing of endogenous genes in resistant cotton via targeting a fragment of the Gbve1 gene compromised cotton resistance to V. dahliae. Furthermore, we transformed the Gbve1 gene into Arabidopsis and upland cotton through Agrobacterium-mediated transformation. Overexpression of the Gbve1 gene endowed transgenic Arabidopsis and upland cotton with resistance to high aggressive defoliating and non-defoliating isolates of V. dahliae. And HR-mimic cell death was observed in the transgenic Arabidopsis. Our results demonstrate that the Gbve1 gene is responsible for resistance to V. dahliae in island cotton and can be used for breeding cotton varieties that are resistant to Verticillium wilt.     

陆地棉主要产量相关性状的SSR标记关联分析

高产优质育种是我国棉花育种的主要目标。寻找与目标性状关联的分子标记,可克服常规育种的盲目性,提高分子标记辅助选择育种的准确性。本研究对118份陆地棉种质资源的衣分、单铃重、单株铃数及子指等4个产量相关性状进行2年2点的表型鉴定,并利用覆盖全基因组的、有多态性的214对SSR标记进行标记与性状的关联分析。结果表明:118份材料的4个产量相关性状表型变异丰富,平均变异系数的变幅在6.1%~19.1%之间,且在各环境中表现较为稳定;基因型分析表明,214对标记共检测到460个等位变异,基因多样性指数平均为0.5151,PIC值平均为0.4587,表明该批标记具有较多的等位变异数和较高的基因多样性;群体结构分析表明该批材料可分为4个亚群,且各类群中材料与地理来源无对应关系;关联分析结果显示,在显著条件下(-log10P1.3,P0.05),共有39个标记位点能够在2个及2个以上的环境中同时检测到,其中有4个标记位点同时与2个以上性状相关联,进一步比较发现,有7个位点与前人研究结果一致,其余32个位点为新发现的位点。研究结果可为陆地棉产量性状遗传改良的分子标记辅助选择提供理论依据。     

New red flower germplasm lines of cotton selected from hybrid of Gossypium hirsutum × G. bickii

By means of dropping GA3 (50 ppm) and NAA (40 ppm) on the hybrid boll-embryo culture in vitro, one F1 plant of G. hirsutum G. bickii was obtained; when F1 branches were grafted on upland cotton and then back-crossed with upland cotton under short-day and cooler-night condition, some BC1 seeds could be harvested. The characteristic segregation was very violent in early generation. Through 3 times of back-crossing and selecting, ten stable hybrid lines with the character of both male parent (viz. red petal-purple spot and strong fibre) and female par-ent (plant type, earliness, white fibre, lint length, etc. ) were established. These lines were assigned as HB red flow-er lines (HBRL). Transference of character of G. bickii to upland cotton was proved to be successful for the first time. These new germplasms may play an important role in both the genetic research and new cotton variety breeding.     

Dynamic infection of Verticillium dahliae in upland cotton

• Verticillium wilt, an infection caused by the soilborne fungus Verticillium dahliae, is one of the most serious diseases in cotton. No effective control method against V. dahliae has been established, and the infection mechanism of V. dahliae in upland cotton remains unknown.
• GFP‐tagged V. dahliae isolates with different pathogenic abilities were used to analyse the colonisation and infection of V. dahliae in the roots and leaves of different upland cotton cultivars, the relationships among infection processes, the immune responses and the resistance ability of different cultivars against V. dahliae.
• Here, we report a new infection model for V. dahliae in upland cotton plants. V. dahliae can colonise and infect any organ of upland cotton plants and then spread to the entire plant from the infected organ through the surface and interior of the organ.
• Vascular tissue was found to not be the sole transmission route of V. dahliae in cotton plants. In addition, the rate of infection of a V. dahliae isolate with strong pathogenicity was notably faster than that of an isolate with weak pathogenicity. The resistance of upland cotton to Verticillium wilt was related to the degree of the immune response induced in plants infected with V. dahliae. These results provide a theoretical basis for studying the mechanism underlying the interaction between V. dahliae and upland cotton. These results provide a theoretical basis for studying the mechanism underlying the interaction between V. dahliae and upland cotton.

     

海岛棉和陆地棉叶片光合能力的差异及限制因素

研究海岛棉和陆地棉两个栽培种间叶片的光合特性及组织解剖结构特性,揭示不同棉花栽培种叶片的光合能力的差异,探讨如何进一步提高海岛棉的光合物质生产能力,以期为高光效棉花品种选育和高产高效栽培实践提供理论基础。在新疆气候生态条件下,选用北疆棉区有代表性的海岛棉品种新海22号和陆地棉品种新陆早13号为试验材料,分别测定了两棉花栽培种叶片的净光合速率（PN）、气孔导度（Gs）及叶片温度的日变化,PN-PPFD（光量子通量密度）响应曲线,PN-Ci（胞间CO2浓度）响应曲线以及叶绿素含量、叶片面积、比叶重和地上部生物量等指标,并观察了叶片的形态解剖结构特性。研究结果表明,当新陆早13号叶片被固定于水平方向上以后,日进程中新海22号的叶片温度显著高于新陆早13号。在上午和下午光强较低时,新海22号和新陆早13号的PN和Gs无明显差异;12:00h至16:00h（北京时间,下同）,新海22号叶片的PN和Gs均显著低于新陆早13号。新海22号和新陆早13号叶片的PN-Ci响应曲线无明显差异。新海22号叶片的Pmax（最大光合速率）低于新陆早13号,而两者的AQY（表观量子效率）无显著差异。新海22号叶片的栅栏组织厚度、比叶重和生物量均显著低于新陆早13号,而叶绿素含量和叶片面积均显著高于新陆早13号。因此,田间条件下,海岛棉叶片的实际光合能力低于陆地棉,但两者具有相似的光合潜力。气孔导度是导致海岛棉和陆地棉叶片实际光合能力存在差异的重要原因,而栅栏组织较薄限制了海岛棉叶片光合潜力的进一步发挥。     

新型标记SRAP在棉花F2分离群体及遗传多样性评价中的适用性分析

将新型分子标记SRAP(Sequence-related Amplified Polymorphism)应用于棉花的遗传研究,并建立了完整的PCR反应体系,此体系稳定可靠、扩增效果好、可重复性强。采用30个SRAP引物组合对海岛棉品种“Pima90”和陆地棉品种“邯郸208”进行比较扩增,29个引物组合可以获得多态性扩增,显示了较高的多态性。对上述两个品种的F2群体进行检测,共产生149个多态性条带,平均每个组合产生5．14个,单引物组合最多可产生13个多态性条带。用SRAP标记对11份陆地棉材料进行遗传多样性检测,30个引物组合中15个组合有多态性,得到22个多态性条带,显示了较高的多态性比率。研究结果表明,SRAP标记可在棉花分子生物学领域中广泛应用。     

陆地棉GST基因家族全基因组分析

谷胱甘肽转移酶(glutathione-S-transferase, GST)是一种普遍存在的具有多功能的超家族蛋白,在植物初次生代谢、逆境胁迫、胞间信号传递等方面具有重要作用;同时,作为配体其在植物激素代谢以及物质转运方面也发挥作用。为了解析陆地棉(Gossypium hirsutum L.) GST基因家族的信息,本研究对该基因家族成员的种类、进化关系、物理定位、基因结构和保守基序以及表达模式进行了分析。结果显示,在陆地棉全基因组中共含有70个GST基因,进化树和基因结构分析将该家族分为U族、F族、T族、Z族、EF1Bγ族和TCHQD族。基因定位分析发现,除了AD/At2、AD/At4、AD/At5、AD/Dt5、AD/Dt10号染色体上没有GST基因外,其他染色体上都有GST基因,并且在AD/At9、AD/Dt7、AD/Dt12、AD/Dt13这4条染色体上出现基因簇。对F族(Phi类) 9个GST基因进行荧光定量分析,结果表明,除GhGSTF1可能为假基因外,GhGSTF2~9等8个基因在陆地棉根、茎、叶以及各个发育时期的纤维中均有表达;结合生物信息学分析,推测GhGSTF8可能参与原花青素/花青素的转运和积累;GhGSTF4、GhGSTF6和GhGSTF9可能在调节陆地棉的生长和胁迫反应中起作用,而GhGSTF2、GhGSTF3、GhGSTF5和GhGSTF7的功能还有待进一步研究。本研究为陆地棉GST基因家族的分子进化及功能研究提供了理论依据。     

广西南部宿生陆地棉的产量构成因素及其通径分析

棉花宿生栽培对于雄性不育系的多年生保持和杂交制种,固定杂种优势、加速杂种优势利用和保存种质资源具有重要意义。探讨陆地棉宿生栽培的产量构成因素及其对产量形成的影响,为陆地棉宿生栽培提供理论依据。采用随机区组设计,对三年生、二年生宿生陆地棉的产量构成因素及其对产量形成的影响进行比较研究。结果表明:与一年生棉比较,宿生棉单位面积总铃数明显增多,多数品种的单铃重无明显变化,衣分变化因品种而异。三年生和二年生各因素对产量影响的趋势一致,总铃数影响最大,衣分影响次之,两者主要是直接影响;单铃重通过总铃数的间接影响较大。一年生各因素对产量影响略有不同。宿生栽培的总铃数明显增多,是增产的主要原因,同时应选择衣分高、受宿生年限影响小的品种。     

High-resolution sequencing of nine elite upland cotton cultivars uncovers genic variations and breeding improvement targets

Structural variations (SVs) are critical factors affecting genome evolution and important traits. However, identification results and functional analyses of SVs in upland cotton are rare. Here, based on the genetic relationships, breeding history and cumulative planting area of upland cotton in China, nine predominant cultivars from the past 60 years (1950s–2010s) were selected for long read sequencing to uncover genic variations and breeding improvement targets for this crop. Based on the ZM24 reference genome, 0.88–1.47 × 10⁴ SVs per cultivar were identified, and an SV set was constructed. SVs affected the expression of a large number of genes during fiber elongation, and a transposable element insertion resulted in the glandless phenotype in upland cotton. Six widespread inversions were identified based on nine draft genomes and high-throughput chromosome conformation capture data. Multiple haplotype blocks that were always associated with aggregated SVs were demonstrated to play a pivotal role in the agronomic traits of upland cotton and drove its adaptation to the northern planting region. Exotic introgression was the source of these haplotype blocks and increased the genetic diversity of upland cotton. Our results enrich the genome resources of upland cotton, and the identified SVs will promote genetic and breeding research in cotton.     

New red flower germplasm lines of cotton selected from hybrid ofGossypium hirsutum XG. bickii

By means of dropping GA₃(50 ppm) and NAA (40 ppm) on the hybrid boll-embryo culturein vitro, one F₁ plant ofG. hirsutum × G. bickii was obtained; when F₁ branches were grafted on upland cotton and then back-crossed with upland cotton under short-day and cooler-night condition, some BC₁ seeds could be harvested. The characteristic segregation was very violent in early generation. Through 3 times of back-crossing and selecting, ten stable hybrid lines with the character of both male parent (viz. red petal-purple spot and strong fibre) and female parent (plant type, earliness, white fibre, lint length, etc.) were established. These lines were assigned as HB red flower lines (HBRL). Transference of character ofG. bickii to upland cotton was proved to be successful for the first time. These new germplasms may play an important role in both the genetic research and new cotton variety breeding.