陆地棉优质纤维渐渗系中外源遗传组分的鉴定与分析

鲁原343是一个渐渗了海岛棉优质纤维基因的陆地棉种质,对其渐渗的优质纤维片段进行鉴定,对利用优质纤维渐渗系改良陆地棉品种的纤维品质具有重要意义。本研究以综合性状优良的转基因抗虫棉鲁棉研22号与鲁原343杂交构建作图群体,利用317对SSR引物对鲁原343和鲁棉研22号进行多态性分析,有24对引物表现多态。利用这些引物进一步和TM-1、优质纤维渐渗片段的供体Ash imoun i作比较,初步鉴定出10个SSR位点与海岛棉渐渗有关。利用这些标记分析(鲁棉研22×鲁原343)F2群体的标记基因型和纤维品质性状的关系,6个标记与纤维品质显著相关,涉及到4条染色体。其中与伸长率相关的标记BNL2986(R2=5.87%)和与长度、细度相关的标记NAU751(R2=6.62%,6.01%)同位于16号染色体的连锁群LG1上,标记间距离为17.7 cM;与纤维成熟度相关的标记BNL3590(R2=8.62%)和与成熟度、伸长率相关的标记BNL3971(R2=15.0%,9.79%)位于2号染色体的连锁群LG3上,标记间距离为4.5 cM;与纤维强度相关的标记BNL3279(R2=8.12%)和与细度相关的标记BNL827(R2=13.94%)分别位于LGD02和25号染色体上。     

Molecular Cloning and Characterization of a Cytokinin Dehydrogenase Gene from Upland Cotton (Gossypium hirsutum L.)

Cytokinins are plant hormones that play crucial roles in plant growth and development. Cytokinin dehydrogenase (CKX), regarded as a main negative regulator in cytokinin metabolism in plants, irreversibly degrades cytokinins into adenine/adenosine moiety. A CKX homologous gene, designated GhCKX, was cloned from upland cotton (Gossypium hirsutum L.). Transgenic tobacco plants over-expressing GhCKX showed a typical cytokinin-deficient phenotype, while CKX-silenced tobacco plants exhibited cytokinin over-producing phenotype. Tissue specifically enhancing the expression of GhCKX in the ovule epidermis of transgenic cotton led to a significant decrease of trans-zeatin and trans-zeatin riboside contents in the ovule. The decline of cytokinins resulted in a significant decrease in fiber initials on a single ovule. Our results indicate that GhCKX encodes a functional CKX, and cytokinins may be required for the initiation of cotton fiber cells.     

Molecular Tagging and Mapping of Quantitative Trait Loci for Lint Percentage and Morphological Marker Genes in Upland Cotton

Using 219 F2 Individuals developed by crossing the genetic standard line TM-1 and the multiple dominant marker line T586 In Gossyplum hirsutum L., a genetic linkage map with 19 linkage groups was constructed based on simple sequence repeat （SSR） markers. Compared with our tetraploid backboned molecular genetic map from a （TM-1xHal 7124）xTM-1 BC1 population, 17 of the 19 I｜nkage groups were combined and anchored to 12 chromosomes （sub-genomes）. Of these groups, four morphological marker genes In T586 had been mapped Into the molecular linkage map. Meanwhile, three quantitative trait loci for lint percentage were tagged and mapped separately on the A03 linkage group and chromosome 6.     

利用染色体片段置换系定位水稻落粒性主效QTL

水稻落粒性是与其生产密切相关的重要性状之一。以7个染色体片段置换系为材料, 采用重叠群代换作图法对控制落粒性的2个主效QTL进行定位。结果表明, 104个SSR标记在亲本间具有多态性, 多态率为68.0%; 4个置换系的落粒性与亲本日本晴的落粒性相似, 表现难落粒。3个置换系与亲本93-11的落粒性相似, 表现易落粒; 7个染色体片段置换系在第1和第6染色体上检出7个置换片段, 其长度分别为23.6、16.5、 6.6、 9.9、 10.4、 20.2和7.1 cM; qSH-1-1被定位在第1染色体RM472－RM1387之间, 遗传距离约为6.6 cM。qSH-6-1为新发现的落粒性主效QTL, 被定位在第6染色体RM6782－RM3430之间,遗传距离约为4.2 cM。利用染色体片段置换系能准确地定位水稻落粒性QTL, qSH-1-1与qSH-6-1的鉴定和初步定位为其进一步的精细定位、图位克隆及分子标记辅助选择奠定了基础。     

利用染色体片段置换系定位水稻落粒性主效QTL

水稻落粒性是与其生产密切相关的重要性状之一。以7个染色体片段置换系为材料,采用重叠群代换作图法对控制落粒性的2个主效QTL进行定位。结果表明,104个SSR标记在亲本间具有多态性,多态率为68.0%;4个置换系的落粒性与亲本日本晴的落粒性相似,表现难落粒。3个置换系与亲本93-11的落粒性相似,表现易落粒;7个染色体片段置换系在第1和第6染色体上检出7个置换片段,其长度分别为23.6、16.5、6.6、9.9、10.4、20.2和7.1 cM;qSH-1-1被定位在第1染色体RM472-RM1387之间,遗传距离约为6.6 cM。qSH-6-1为新发现的落粒性主效QTL,被定位在第6染色体RM6782-RM3430之间,遗传距离约为4.2 cM。利用染色体片段置换系能准确地定位水稻落粒性QTL,qSH-1-1与qSH-6-1的鉴定和初步定位为其进一步的精细定位、图位克隆及分子标记辅助选择奠定了基础。     

Partial Dominance,Overdominance and Epistasis as the Genetic Basis of Heterosis in Upland Cotton (Gossypium hirsutum L.)

Determination of genetic basis of heterosis may promote hybrid production in Upland cotton (Gossypium hirsutum L.). This study was designed to explore the genetic mechanism of heterosis for yield and yield components in F_{2: 3} and F_{2: 4} populations derived from a hybrid ‘Xinza No. 1’. Replicated yield field trials of the progenies were conducted in 2008 and 2009. Phenotypic data analyses indicated overdominance in F₁ for yield and yield components. Additive and dominance effects at single-locus level and digenic epistatic interactions at two-locus level were analyzed by 421 marker loci spanning 3814 cM of the genome. A total of 38 and 49 QTLs controlling yield and yield components were identified in F_{2: 3} and F_{2: 4} populations, respectively. Analyses of these QTLs indicated that the effects of partial dominance and overdominance contributed to heterosis in Upland cotton simultaneously. Most of the QTLs showed partial dominance whereas 13 QTLs showing overdominance in F_2:3 population, and 19 QTLs showed overdominance in F_2:4. Among them, 21 QTLs were common in both F_{2: 3} and F_{2: 4} populations. A large number of two-locus interactions for yield and yield components were detected in both generations. AA (additive × additive) epistasis accounted for majority portion of epistatic effects. Thirty three complementary two-locus homozygotes (11/22 and 22/11) were the best genotypes for AA interactions in terms of bolls per plant. Genotypes of double homozygotes, 11/22, 22/11 and 22/22, performed best for AD/DA interactions, while genotype of 11/12 performed best for DD interactions. These results indicated that (1) partial dominance and overdominance effects at single-locus level and (2) epistasis at two-locus level elucidated the genetic basis of heterosis in Upland cotton.     

Mapping and genomic targeting of the major leaf shape gene (L) in Upland cotton (Gossypium hirsutum L.)

Key message

A major leaf shape locus (L) was mapped with molecular markers and genomically targeted to a small region in the D-genome of cotton. By using expression analysis and candidate gene mapping, two LMI1 -like genes are identified as possible candidates for leaf shape trait in cotton.

Abstract

Leaf shape in cotton is an important trait that influences yield, flowering rates, disease resistance, lint trash, and the efficacy of foliar chemical application. The leaves of okra leaf cotton display a significantly enhanced lobing pattern, as well as ectopic outgrowths along the lobe margins when compared with normal leaf cotton. These phenotypes are the hallmark characteristics of mutations in various known modifiers of leaf shape that culminate in the mis/over-expression of Class I KNOX genes. To better understand the molecular and genetic processes underlying leaf shape in cotton, a normal leaf accession (PI607650) was crossed to an okra leaf breeding line (NC05AZ21). An F₂ population of 236 individuals confirmed the incompletely dominant single gene nature of the okra leaf shape trait in Gossypium hirsutum L. Molecular mapping with simple sequence repeat markers localized the leaf shape gene to 5.4 cM interval in the distal region of the short arm of chromosome 15. Orthologous mapping of the closely linked markers with the sequenced diploid D-genome (Gossypium raimondii) tentatively resolved the leaf shape locus to a small genomic region. RT-PCR-based expression analysis and candidate gene mapping indicated that the okra leaf shape gene (L ^o ) in cotton might be an upstream regulator of Class I KNOX genes. The linked molecular markers and delineated genomic region in the sequenced diploid D-genome will assist in the future high-resolution mapping and map-based cloning of the leaf shape gene in cotton.     

棉花高频体细胞胚胎发生及再生体系建立

棉花的体细胞胚胎发生率低,限制了转基因棉花的发展。为了扩大可再生棉花的基因型,发展新疆优质棉花的特点,以新疆的主要栽培品种新陆早33为材料,以下胚轴为外植体,使用葡萄糖,麦芽糖作为主要碳源,用Phytagel固化,对不同激素的浓度组合和培养条件进行探索,对棉花胚性愈伤的诱导到体细胞胚胎的发生阶段进行优化,体细胞胚胎成熟以后进行萌发培养可以获得正常的植株。通过试验证明,有利于棉花的愈伤组织生长的激素浓度为KT 0.1 mg/L,2,4-D 0.1 mg/L,下胚轴的中部诱导愈伤形态最好。体细胞胚胎发生阶段以KT,IBA组合,以低盐培养基进行子叶胚的成苗,建立了再生体系。     

陆地棉叶绿体铜锌超氧化物歧化酶基因的克隆与表达

以陆地棉‘CRI36＇的叶片为材料,使用RACE技术克隆到了棉花叶绿体Cu/Zn-SOD酶基因。基因序列全长共1 043 bp,含有完整的开放阅读框。推导的氨基酸序列分析显示含有叶绿体信号肽,和已知植物的叶绿体Cu/Zn-SOD酶蛋白的氨基酸残基的同源性在66%～74%之间。基因的表达谱分析显示：棉花叶绿体Cu/Zn-SOD酶基因主要在叶片、茎中表达,根、花和下胚轴中没有检测到信号,即基因的表达主要在棉花的绿色组织。不同生育期的表达谱结果证实：该基因主要在苗期表达,以后表达逐渐减少。用pET-21a（＋）构建了原核表达载体,在大肠杆菌BL21（DE3）的表达结果显示：表达后得到一个29.0 kD的新蛋白,其分子量与预期目标一致。对SOD酶活性的分析证实,重组菌的酶活性显著增加,证明克隆的基因具有活性。     

陆地棉新型核雄性不育系21A的初步研究

利用辐射诱变和回交育种成功培育出21A雄性不育系,经遗传分析和等位性测验发现,该不育性受1对隐性基因控制,它与国内发现的msc1、msc2、msc3、msc74个陆地棉隐性不育基因彼此是非等位基因;采用NCⅡ遗传交配设计,用21A不育系作母本,5个常规棉品种作父本,配制杂交组合,对10个组合的F1产量和品质性状的竞争优势和配合力进行分析,结果表明,该不育系杂种优势明显,一般配合力高,易于筛选优良的杂交组合,在生产优质杂交种方面具有很大的利用价值。     

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。     

矮秆陆地棉品种陆矮1号的生长发育特性研究

本研究以矮秆陆地棉品种陆矮1号为材料,以株高正常品系97004H和陆地棉遗传标准系TM-1为对照,通过田间试验,比较研究了不同品种的生长发育规律。结果表明：陆矮1号苗期形态与对照品种无显著差异,随着生育进程株高差异逐渐增大,现蕾后主茎日增长量极显著低于对照品种,日增长高峰与对照品种一致,但高峰期的日增长量则不到对照品种的一半,盛花期以后主茎基本停止生长;最终株高只有对照品种的1／2左右。单株果枝数及主茎伸长节间数显著少于97004H和TM-1,每果枝的果节数及果枝长度与对照相比有极显著差异,主茎第12节以下节间长度与对照品种无显著差异,上部节间显著短于对照品种。株高差异主要发生在盛蕾期以后,由中上部节间缩短引起。陆矮1号生育期较短,结铃部位和成铃时间集中。     

水稻外观品质的数量性状基因位点分析

利用由98个家系组成的Nipponbare(粳)／Kasalath(秒)∥Nipponbare回交重组自交系(backcross inbred lines,BILs)群体(BC1F9)及其分子连锁图谱,采用复合区间作图的方法,在2个不同年份对粒长、粒宽、粒形、垩白率、垩白大小、垩白度和透明度等7个稻米外观品质性状的数量性状基因位点(Quantiative trait loci,QTL)进行了定位分析。共定位到33个四QTLs,单个性状QTL数目在4-7个之间,以垩白率最多,为7个;粒长和垩白大小次之,为5个;其他性状均为4个,表明该组合外观品质是由多基因控制的数量性状。单个QTL对性状变异解释率粒长为6．2％-15．2％,粒宽为8．3％-32．5％,长宽比为6．8％-19．8％,垩白率为6．4％-28．5％,垩白大小为6．1％-16．9％,垩白度为9．3％-17．2％,透明度为5．6％-25．2％．QTL在染色体上成集中分布的特点,第3染色体C1488-C563、第5染色体R830-R3166和R1436-R2289、第6染色体R2147-R2171均有3个以上的QTLs分布。比较2年的检测结果表明,外观品质性状的QTL定位都受环境影响,但不同性状受影响的程度差异很大。粒长和粒形的QTL定位受环境影响很小,垩白率、垩白大小和垩白度的QTL定位受环境影响很大。     

水稻粒长QTL定位与主效基因的遗传分析

该研究利用短粒普通野生稻矮杆突变体和长粒栽培稻品种KJ01组配杂交组合F_1,构建分离群体F_2;并对该群体粒长进行性状遗传分析,利用平均分布于水稻的12条染色体上的132对多态分子标记对该群体进行QTL定位及主效QTLs遗传分析,为进一步克隆新的主效粒长基因奠定基础,并为水稻粒形育种提供理论依据。结果表明:(1)所构建的水稻杂交组合分离群体F_2的粒长性状为多基因控制的数量性状。(2)对543株F_2分离群体进行QTL连锁分析,构建了控制水稻粒长的连锁遗传图谱,总长为1 713.94 cM,共检测出24个QTLs,只有3个表现为加性遗传效应,其余位点均表现为遗传负效应。(3)检测到的3个主效QTLs分别位于3号染色体的分子标记PSM379～RID24455、RID24455～RM15689和RM571～RM16238之间,且三者对表型的贡献率分别为54.85%、31.02%和7.62%。(4)在标记PSM379～RID24455之间已克隆到的粒长基因为该研究新发现的主效QTL位点。     

棉花高品质纤维性状的主基因与多基因遗传分析

利用主基因与多基因混合遗传模型联合分析方法 ,通过纤维强度不同的 5个亲本配制的 8个组合 ,研究了棉花主要纤维品质性状的遗传。联合分析发现 ,在不同性状不同组配方式的 14个组合中 ,有 12个存在主基因 ,表明了纤维性状主基因存在的普遍性 ,以F2∶3 家系的预测效果最好 ;双亲纤维品质性状均存在较大差异的组合——— 72 35×TM1F2 代强度主基因的遗传率为 0 .196 ,麦克隆值为 0 .32 0 ,长度为 0 .139,回交世代的主基因遗传率小。除纤维长度总的显性效应为较高的正值外 ,其余各纤维性状的主基因显性与多基因显性的总和为负值或接近 0 ,杂合状态下大多数纤维品质性状表型值会偏向中亲值或低亲值 ,单纯依靠表型选择效率低。因此 ,很有必要对棉花品质性状进行分子标记辅助育种选择     

陆地棉核不育株扦插繁殖与宿生保持及杂种优势利用

利用广西南宁冬季无霜冻的气候特点,对陆地棉(Gossypium hirsutum L.)洞A细胞核雄性不育扦插株与宿生株及其杂交一代进行比较研究.结果表明:(1)1 a生扦插株开花较早,茎粗、主茎节间长度、铃重、子指4个性状显著好于实生株,但扦插株的种子产量显著低于实生株,其原因是果枝扦插形成的僵苗占扦插株的23.04%,而且扦插株全为不育株,实生株中则有50%左右的可育株;(2)2 a生扦插株性状与实生株无显著差异;(3)由扦插繁殖的不育株和种子繁殖的不育株配制的杂交F1代的产量与纤维品质无显著差异.因此认为,扦插繁殖陆地棉核不育株用于多年杂交制种是可行的,选择营养枝扦插是陆地棉核不育株扦插繁殖需要注意的关键问题. 宿生株及其杂交一代进行比较研究.结果表明:(1)1 a生扦插株开花较早,茎粗、主茎节间长度、铃重、子指4个性状显著好于实生株,但扦插株的种子产量显著低于实生株,其原因是果枝扦插形成的僵苗占扦插株的23.04%,而且扦插株全为不育株,实生株中则有50%左右的可育株;(2)2 a生扦插株性状与实生株无显著差异;(3)由扦插繁殖的不育株和种子繁殖的不育株配制的杂交F1代的产量与纤维品质无显著差异.因此认 ,扦插繁殖陆地棉核不育株用于多年杂交制种是可行的,选择营养枝扦插是陆地棉核不育株扦插繁殖需     

陆地棉产量性状QTLs的分子标记及定位

用我国的高产栽培品种泗棉3号和美国栽培品种TM-1为材料,构建F₂和F_2∶3作图群体,应用301对SSR引物和1040个RAPD引物,对产量性状QTLs进行了分子标记筛选,结果共筛选出了37对SSR多态性引物和10个RAPD多态性引物的49个位点,鉴定出了控制产量性状变异的主效QTLs。定位于第9染色体的连锁群,分别具有控制铃重、衣分和籽指的主效QTLs,铃重的2个QTLs分别解释F_2∶3群体表型变异的18.2%和21.0%;在F₂群体检测到的1个衣分QTL解释表型变异的25%,另一个衣分QTL在F₂群体和F_2∶3群体都检测到,解释F₂群体衣分的24.9%的表型变异,解释F_2∶3群体衣分的5.9%的表型变异;在F_2∶3群体铃重的一个QTL的同一位置同时检测到一个籽指QTL,它解释15.6%的表型变异,是一因多效或是紧密连锁的两个QTLs,有待进一步研究。本研究标记的产量性状主效QTLs可用于棉花产量性状的标记辅助选择。     

Mapping of Quantitative Trait Loci for Butter Content and Hardness in Cocoa Beans (Theobroma cacao L.)

Cocoa butter is an important raw material for the chocolate, pharmaceutical, and cosmetic industries. The butter content and quality in cocoa beans are genetically controlled characteristics, and affect its commercial value and industrial applicability. In the present work, an F₂ population derived from the cross between the ICS-1 and Scavina-6 cocoa clones was used for molecular mapping. A linkage map was constructed based on amplified fragment length polymorphism, random amplified polymorphic DNA, and simple sequence repeat markers, resulting in a total of 273 markers, distributed in 14 linkage groups (LGs). Phenotyping of butter content was performed after ether extraction and butter hardness was determined by sweeping differential calorimetry. One quantitative trait locus (QTL) associated to butter content was mapped at linkage group 9 (LG9) and two QTLs for butter hardness were identified at linkage groups 9 and 7 (LG9 and LG7). The two QTLs mapped at the LG9 explained 51.0% and 28.8% of the phenotypic variation for butter content and hardness, respectively. These QTLs were concentrated in the same map region, suggesting a close genetic linkage or pleiotropic effect. The QTLs identified may be useful in further marker-assisted selection breeding programs aimed at cocoa butter quality improvement.