陆地棉与澳洲2个野生棉杂种细胞学研究

本文首次报道了陆地棉×南岱华棉杂种Ｆ１的细胞学研究结果，并与陆地棉Ｘ斯特提棉的杂种进行了对比分析。供分析的细胞数均在７００个以上，中期Ⅰ染色体构型种类占总ＰＭＣ的百分率（构型率）均在７％以下，陆地棉与斯特提棉和南岱华棉杂种Ｆ１的染色体平均构型分别为２１．９７Ⅰ＋７，９７Ⅱ＋０．３５Ⅲ＋０．０１Ⅳ和２０．６２Ⅰ＋８．３７Ⅱ＋０．５２Ⅲ＋０．０２Ⅳ，单一细胞环状二价体、染色体桥和二价体交叉率分别是０．５３与０．４８、７．４４与７．８９和１．０７及１．０６。在２个杂种中均观察到八价体，但为数极少。中期Ⅰ时期二价体和多价体集中于细胞赤道面上，而单价体分散在周围。后期Ⅰ及以后，单价体分布完全随机，形成多个染色体块或片段。四分体形成时期，小孢子数目和大小及形状变异均较大。主要表现在小孢子不是规则的团体，体积大小相差可达２～４倍，单一ＰＭＣ中的数目从ｌ至１３，但其四分体占多数，均在６０％以上。且微核变幅较大，０～６个。经过比较分析，说明两个棉种亲缘关系很近，与陆地棉的种间关系差异也不大。     

四倍体栽培棉与索马里棉异源五倍体杂种的细胞遗传学研究

花粉母细胞(PMC)压片观察{[陆地棉(G.hirsutum L)×索马里棉(G.somalense Grke)]×海岛棉(G.barbadense)}的三元杂种及[(陆地棉×索马里棉)×陆地棉]的二元杂种中期Ⅰ染色体构型,这两个杂种都是异源五倍体(2n=5x=65),比较了两个杂种形成的多价体、二价体及单价体的频率。三元杂种具有多价体的PMC占22.03％,具有棒状二价体的PMC占57.29％,而二元杂种分别为8.33％和31.74％。三元杂种形成异源多价体和棒状二价体的频率比二元杂种高一倍。以上结果暗示在三交种的异源五倍体减数分裂中,存在同源染色体与部分同源染色体竞争联会现象,进而增加了异源易位的频率,有利于E_2染色体组基因渐渗到AD染色体组。     

球茎大麦在大麦育种上的应用研究

本文介绍栽培大麦(2x)与球茎大麦(4x)种间杂种的产生及其细胞学鉴定结果。二者杂交成胚率平均可达48.43%,胚培成苗率平均可达25.64%。杂种F1染色体数为21,通过C-带鉴定表明,7条为母本栽培大麦染色体、14条为父本球茎大麦染色体。F1花粉母细胞中染色体构型:单价体4-7个、三价体1-3个,其余为二价体。杂种植株形态为父本球茎     

大豆雄性不育突变体NJ89-1的细胞学研究

通过对大豆雄性不育突变体NJ89-1不育株与可育株小 孢子母细胞减数分裂的比较研究,发现NJ89-1雄性不育株小孢子母细胞减数分裂高度变异,联会异常,出现单价体、染色体落后、染色体桥、染色体不均等分离等;在形成四分体的早期阶段,小孢子母细胞里见到微核,以后产生大小不等的二分体、三分体、四分体及多分体;单核小孢子     

特异同源三倍体水稻材料SAR-3细胞学研究

以特异同源三倍体水稻材料SAR-3中15个株系为研究材料, 对同源三倍体减数分裂中染色体行为, 特别是染色体分离方式进行了研究. 结果表明: 同源三倍体SAR-3花粉母细胞减数分裂中期Ⅰ染色体形成单价体、二价体和三价体; 后期Ⅰ染色体分离遵守随机分配规律, 但有一定比例整倍性二分体出现; 后期Ⅰ、后期Ⅱ染色体落后是形成单倍性     

抗白粉病普通小麦-簇毛麦新种质的遗传学及生化鉴定

普通小麦-簇毛麦抗白粉病新种质94G22-1和94-G33-1都对白粉病免疫。其体细胞染色体数为42,花粉母细胞减数分裂中期Ⅰ染色体构型为21个二价体。用感病小麦品种对其测交,F_1代全部对白粉病表现免疫,F_1代花粉母细胞减数分裂中期Ⅰ染色体构型为20个二价体加2个单价体。测交F_2抗感比符合3:1的预期分离比例。体细胞快速荧光原位杂交表明94G22-1、94G33-1都带有两条簇毛麦染色体;染色体C-分带证明这两条簇毛麦染色体是一对6V;重双端体分析确认6V代换的普通小麦染色体是6D。在谷草转氨酶(GOT)同工酶电泳图潜的GOT-2活性区,94G22-1、94G33-1显示了3条酶带,其中1条来自簇毛麦,由6V上的基因编码,亚基组成分析亦确证94G22-1、94G33-1是6D(6V)异代换系。     

四倍体栽培棉与二倍体野生棉杂种PMC-FISH研究

 首次将PMC-FISH（花粉母细胞荧光原位杂交）技术应用于四倍体栽培棉与二倍体野生棉杂交所形成的三倍体杂种棉F₁中,成功的鉴定了目标染色体中的单价体、双价体、多价体,还发现在非目标染色体上有星状和片段状的杂交信号。在以A组棉基因组DNA作为探针的PMC-FISH中,分别对三倍体杂种棉F₁及其母本四倍体栽培棉的减数分裂中期I的染色体构型进行了鉴定。结果显示,四个三倍体杂种棉F₁的减数分裂时期染色体的构型分别是：陆地棉(AD)₁×雷蒙德氏棉（D₅）为1IIIaad + 1IIaa + 4IIad + 7IIdd + 5Ⅰa + 7Ⅰd;海岛棉(AD)₂×旱地棉（D₄）为1Ⅴaaaad +1IIIadd + 2IIad + 8IIdd + 6Ⅰa + 5Ⅰd;陆地棉(AD)₁×澳洲棉（C₃）为2IIIadd (adc or acc) + 1IIaa + 9Ⅰa + 4IIcc (dc or dd) + 14Ⅰd (c);陆地棉(AD)₁×南岱华棉（C_1-n）为：1IIaa + 1IIad (ac)+ 10Ⅰa + 6IIcc (dc or dd) + 13Ⅰd (c)。然而,两个四倍体棉染色体的构型都是13IIaa + 13IIdd。上述结果还显示, AD×D的两个杂种组合中,二价体多,单价体少,AD×C的两个杂种组合中,二价体少,单价体多;并且AD×D型杂种中A亚组染色体单体的数目比其在AD×C型杂种中的更少。这说明,与C染色体组相比较,D染色体组与四倍体棉种的亲缘关系更近。同时,我们的结果也证明PMC-FISH技术在分析三倍体杂种棉F1的亲缘关系中起着不可取代的作用。     

三元杂种及回交后代的遗传性状和细胞学研究

三元杂种[陆地棉×(亚洲棉×比克氏棉)]的回交后代BC4F1发生疯狂分离,在各个性状上都表现出特殊的变异类型,在育性上分离出三种类型可育株,部分可育株,不育株。对这三种类型和三元杂种F1花粉母细胞减数分裂的观察表明,三元杂种F1及回交后代的不育株减数分裂表现异常,大多染色体不能联会配对,以单价体的形式存在,且有落后染色体、三价体、四体环的出现,在末期形成了不正常的四分孢子和多分孢子,这是不育的主要原因;部分可育株,大部分染色体联会配对成棒状二价体,个别为单价体;可育株染色体配对正常,能形成可育的花粉粒。     

(亚洲棉×比克氏棉)F1双二倍体不育系的花粉败育机理研究

用亚洲棉与比克氏棉进行杂交,对杂种F1进行染色体加倍,获得(亚洲棉×比克氏棉)F1双二倍体。(亚洲棉×比克氏棉)F1双二倍体后代中分离出可育和不育两种类型。不育系形态特征与可育系相似,整体性状似父本比克氏棉,多数花为闭花授粉。花粉母细胞减数分裂染色体观察结果表明,不育系与可育系细胞的染色体数目相同,但不育系的单价体较多,染色体构型为2n=52=7.07I+18.13II+2.00III+0.67IV,二价体平均交叉数为1.91;可育系的染色体构型为2n=52=3.09I+23.79II+0.39III+0.04IV,二价体平均交叉数为1.91。另外,不育虽能形成明显的赤道板,但异常的纺缍体使在赤道板上的染色体方向各异,后期I染色体分向两极数目不均等,一些配对染色体成整体运动,而不分向两极。中期II中出现染色体不同步。后期II出现各种畸形孢子群,结果产生各类不育花粉粒。     

四种栽培棉合成的四元杂种F1细胞遗传学研究

用(亚洲棉×草棉)F1进行染色体加倍,再与(陆地棉×海岛棉)F1进行杂交,产生亚洲棉、草棉、陆地棉和海岛棉四元杂种.对四种栽培棉的四元杂种F1花粉母细胞减数分裂各时期染色体行为观察结果表明,四元杂种F1的减数分裂异常,染色体丢失现象普遍,部分染色体不能联会配对,以单价体的形式存在,并出现三价体、四价体、五价体等多价体,在减数分裂的末期形成大量的不正常多分孢子,导致四元杂种F1高度不育.然而,四元杂种F1减数分裂中大多染色体仍能配对,形成环状或棒状二价体,并有较多的三价体、四价体、五价体等多价体,表明四种栽培棉种之间的亲缘关系相对较近,可以通过遗传重组产生综合四个栽培棉种性状的新种质.     

Investigation on Evolutionary Relationships of the Subgenomes in Interspecific Triploid Cotton via Meiotic FISH

We report in this paper primary studies on interspecific species of cotton vis GISH (genomic in situ hybridization).We use interspecific triploid hybrids (F1 from hybridization of allotetraploid cultivated species with diploid A,D,or C genome species) and two cultivated tetraploids to study the chromosome paring during meiosis of pollen mother cellls (PMCs) and to estimate the consequences on synapsis between these three subgenomes after synthetic polyploid formation.     

Chromosome doubling and fertility study of Alstroemeria aurea × A. caryophyllaea

Self-pollinations of a diploid (2n = 2x = 16) interspecific hybrid Alstroemeria aurea × A. caryophyllaea, resulted in no seeds. Backcrosses of the hybrid with parent A. aurea did not produce any seeds. In an attempt to restore the hybrid fertility, an efficient in vitro procedure has been developed and applied effectively in the chromosome doubling of the diploid hybrid. Forty-one percent of the treated plants were proven to be truly tetraploid by chromosome counts and stomatal measurements after applying 0.2 to 0.6% colchicine for 6 to 24 hours. Over 87.5% of these colchicine-induced tetraploids were stable and retained their tetraploidy after one year of growth. The fertility of the hybrid was not restored although the pollen stainability was increased from 0 to 12% after chromosome doubling. Cytological studies on the pollen mother cells (PMCs) of the sterile diploid hybrids revealed abnormal meiotic behaviors. In addition, aneuploid chromosome numbers, ranging from 2n = 1 to 2n = 18, were observed in over 45% of the PMCs examined. PMCs of the colchicine-induced tetraploids showed that meiotic chromosome pairings were normal in most cases (1.59_I + 15.07_II + 0.05_III + 0.03IV). These results indicate that the sterility of this hybrid is not only caused by parental chromosome differences, but other complex fertility/sterility-regulating mechanisms are involved too. This revised version was published online in July 2006 with corrections to the Cover Date.     

渝棉1号优质纤维QTL的标记与定位

利用陆地棉遗传标准系TM-1和优质品种渝棉1号组建了(TM-1×渝棉1号) F₂和F_2:3分离群体。通过5 544对SSR引物对亲本进行筛选,获得178个多态性标记,用其中138个构建了总长为959.7 cM的遗传图谱,覆盖棉花基因组的19%。应用复合区间作图法分析了该组合的F₂单株和F₃家系纤维品质性状,共检测到12个纤维品质数量性状基因座(QTL),包括1个纤维长度的、4个纤维强度的、3个马克隆值的、3个整齐度的和1个伸长率的,分别解释各性状表型变异的6.1%、5.31%~14.62%、7.88%~19.17%、7.4%~11.71%和8.26%。研究还发现Chr.23和Chr.24是优质纤维QTLs的富集区。     

Genome discrimination and chromosome pairing in the Hordeum chilense × Aegilops tauschii amphiploid

Tritordeum (X Tritordeum Ascherson et Graebner) is a synthetic amphiploid belonging to the Triticeae tribe, which resulted from crosses between Hordeum chilense and wheat. It presents useful agronomic traits that could be transferred to wheat, widening its genetic basis. In situ hybridisation with total genomic DNA from H. chilense and cloned, repetitive DNA sequences (pTa71 and pAs1) probes were used to discriminate the parental origin of all chromosomes, to analyse the chromosome pairing and to identify the chromosomes in pollen mother cells (PMCs) at metaphase I of the tritordeum line HT251 (H^chH^chDD, 2n = 4x = 28). The H. chilense total genomic DNA and the ribosomal sequence pTa71 probes, allowed the unequivocal discrimination of the 14 chromosomes of H^ch genome-origin and the 14 chromosomes of D genome-origin. Chromosome pairing analysis revealed meiotic irregularities such as reduced percentage of PMCs with complete homologous pairing, high frequency of univalents, most of H. chilense-origin and a reduced frequency of intragenomic multivalents from both genomes. The H. chilense genome revealed high meiotic instability. After individual chromosome identification at metaphase I with the pAs1 probe, we found the occurrence of pairing between chromosomes of different homoeology groups. The possible interest of the tetraploid tritordeum in the improvement of other Triticeae species is also discussed.     

偏凸-柱穗山羊草双二倍体SDAU18 PMC MI染色体数目异常

为探讨偏凸-柱穗山羊草双二倍体SDAU18减数第一分裂中期(PMCMI)染色体丢失特点,利用改良卡宝品红压片法,对SDAU18及其双亲的PMCMI染色体构型进行观察。结果表明:在SDAU18的同一枚花药中,部分PMCsMI出现了不同程度的染色体丢失现象,发生染色体丢失的PMCMI频率平均为4.64%,染色体数目异常的PMCsMI的平均染色体构型为:2n=12.77I+5.26IIR+1.57IIO+0.02III=26.49。其双亲的PMCMI的染色体数目均表现正常,均由14个二价体组成。SDAU18PMCSMI染色体数目变异的特点比较罕见。     

棉花纤维特异/优势表达基因的染色体定位

 棉纤维是研究植物细胞伸长和细胞壁建成以及纤维素生物合成的优良模型,迄今为止,已经分离了许多纤维特异/优势表达的基因。为了便于这些基因的图位克隆使其能够应用于棉花纤维品质的改良中,本研究采用分离群体定位法和Blast分析法对这些基因进行染色体定位。利用陆地棉、海岛棉BC₁种间分离群体,将GhCFE定位在第6染色体,GhGLP1-250定位在第19染色体。Blast分析将11个基因定位到棉花染色体上。这些基因与棉纤维的伸长和细胞壁的合成相关,与这些基因连锁标记的获得有助于棉花纤维长度和强度的分子标记辅助选择。     

Seed trait evaluation of <Emphasis Type="Italic">Gossypium barbadense</Emphasis> L. chromosomes/arms in a <Emphasis Type="Italic">G. hirsutum</Emphasis> L. background

Investigation of cotton nutritional components is important because its seeds provide a useful nutritional profile and can possibly serve as a biofuel resource. In this study, five cultivars, 13 cotton chromosome substitution (CS-B) lines, their donor parent, '3-79', and their recurrent parent, 'TM-1', were evaluated for seed traits over four environments. A mixed linear model approach with the jackknife method was employed to estimate variance components and to predict genotypic effects for each seed trait. Genotypic effects were more important than genotype by environment interaction for all seed traits. Chromosome associations with these seed traits were detected using the comparative method by comparing the differences between each CS-B line and TM-1. For example, chromosome 4 of 3-79 in TM-1 background was associated with reduced seed index (SI), embryo percentage, protein percentage while associated with increased seed oil percentage and seed fiber percentage. Other chromosome associations with these seed traits were also observed in this study. SI was highly correlated with three seed index traits: seed protein index, seed oil index (OI), and seed fiber index. Lint percentage, boll number, and lint yield were positively correlated with protein percentage while negatively correlated with SI and OI. SI and seed fiber content exhibited negative correlations with micronaire but positive correlations with fiber length and strength. Results suggested that agronomic traits and seed nutrition components can be improved simultaneously.     

野生棉与陆地棉异源四倍体的合成与性状鉴定

棉属具有丰富的种质资源,将抗病性从野生棉转移到陆地棉中,以此提高陆地棉抗病性是一条理想的育种途经。本研究通过陆地棉和C组野生澳洲棉远缘杂交获得杂种F1,对其进行染色体加倍,将加倍的异源六倍体再与B组野生绿顶棉杂交,产生陆地棉、澳洲棉、绿顶棉的异源四倍体杂种。异源四倍体杂种继承了亲本的部分性状,同时也产生了如蔓生等特异性状;该杂种减数分裂后期染色体不能均等分离,四分体时期有多分体和微核出现;SSR结果发现,该杂种不仅具有三个亲本的特征条带,并且产生了新的重组成分。本研究不仅为该异源四倍体杂种不育提供了直接原因,也为进一步探讨有效的育性恢复方法及棉花新种质创制提供了依据,同时为利用野生棉资源控制棉花黄萎病育种探索培育了中间材料。     

Genetic effects of nine <Emphasis Type="Italic">Gossypium barbadense</Emphasis> L. chromosome substitution lines in top crosses with five elite Upland cotton <Emphasis Type="Italic">G. hirsutum</Emphasis> L. cultivars

Crosses between Gossypium barbadense L and Gossypium hirsutum L. (Upland cotton) have produced limited success in introgressing fiber quality genes into the latter. Chromosome substitution lines (CSBL) have complete chromosomes or chromosome arms from G. barbadense, line 3-79, substituted for the corresponding chromosome or arms in G. hirsutum in a near isogenic background of TM-1. We top crossed nine CSBL and their parents (TM-1 and 3-79) with five cultivars. Parental lines and their F₂ populations were evaluated in four environments for agronomic and fiber quality traits. The CSBL and their F₂ hybrids showed wide ranges for both agronomic and fiber traits of economic importance. Genetic analysis showed that additive variances were larger than dominance variances for lint percentage, boll weight, lint yield, fiber length, strength, elongation, micronaire, and yellowness; whereas, dominance variances were larger than additive variances only for uniformity of fiber length and equal for fiber reflectance. For all traits, except boll weight and lint yield, significant additive effects of one or more chromosomes from 3-79 in TM-1 background were greater than the corresponding TM-1 chromosome. In addition, we identified specific chromosomes from G. barbadense (3-79) that carry alleles for improvements in specific fiber quality traits in Upland cotton. Favorable additive effects of individual chromosomes or chromosome segments from 3-79 relative to corresponding chromosomes or chromosomes segments from TM-1 were identified in this study as follows: Lint percentage, chromosome/arms 10, 16-15; longer fibers, chromosome/arms 01, 11sh, 26Lo; more uniform fibers, chromosomes/arms 01, 11sh, 10, 17-11; stronger fibers, chromosome/arms 01, 11sh, 12sh, 26Lo, 17-11; fiber elongation, chromosomes/arms 01, 11sh, 26Lo, 10, 17-11; reduced fiber micronaire, chromosome/arms 01, 12sh, 4-15, 16-15, 17-11; fibers with more reflectance, chromosome/arms 10, 4-15, 16-15, 17-11; fiber with less yellowness, chromosome arms 4-15, 17-11. Based on the present study, we concluded that by using CSBL, favorable fiber quality alleles can be introgressed into Upland cotton, thus greatly improving the breeder’s ability for improvement of Upland cotton for a variety of traits. These data should provide useful genetic information to the cotton breeding industry at large.