陆地棉子叶色素腺体延缓形成性状的表现与组织结构观察

用(亚洲棉×比克氏棉)F1双二倍体作母本与陆地棉单节显性系1(Gl2Gl2gl3gl3)进行杂交和连续回交,育成了具有子叶色素腺体延缓形成特性的陆地棉种质系———ABH0318。该种质的休眠种子观察不到色素腺体,但解剖镜下可看到子叶边缘处有小量稀小的色素腺体。组织切片观察发现,ABH0318休眠种子内含有3种不同类型的色素腺体结构:(1)具有色素腺雏形但无腺腔的色素腺体原,约占总色素腺体或结构的50%左右;(2)发育不全的色素腺体,其体积较小,腺腔内无内含物,约占25%左右;(3)正常色素腺体类型,分布于子叶的边缘,约占25%左右。种子萌发后,第一类色素腺体结构中的细胞迅速溶解,形成腺腔,并迅速形成正常色素腺体,而第二、三类色素腺体结构无明显的变化。     

有酚与低酚陆地棉亲本配制的种间三元杂种腺体遗传性状研究

本研究对（亚洲棉Ｘ比克氏棉）Ｘ陆地棉种间三元杂种植株和花器形态特性、腺体表现和棉酚含量进行了比较和研究。结果表明，用有酚棉和低酚棉作陆地棉亲本配制的不同类型的种间三元杂种在植株和花器形态性状上无显著差异；但在各器官和组织上的腺体表现和棉酚含量有明显的差异。用低酚棉配制的三元杂种完全表现有比克氏棉子叶腺体延缓发生性状；而用有酚棉配制的三元杂种在种子上尚有少量腺体，但包括花器在内的其它器官和组织均为有腺体类型，腺体密度和大小与一般有酚棉相同。此外，本文对比克氏棉子叶腺体延缓发生性状的遗传和腺体与棉酚含量的关系等问题进行了讨论。     

比克氏棉色素腺体形态建成与棉酚代谢特性的研究

【目的】对比克氏棉(G.bickii Prokh)种子形成过程和种子萌发过程中的色素腺体形态建成特点、色素腺体与棉酚含量的动态关系及其分子机理进行探究。【方法】以比克氏棉为材料,进行组织切片观察、棉酚含量测定并分析对种子萌发期色素腺体形成及棉酚合成基因的表达谱。【结果】比克氏棉在种胚形成过程中,子叶上形成大量色素腺体原,并在种子萌发后12 h形成腔体结构,转换成成熟的色素腺体;比克氏棉种胚形成过程及种子萌发过程中均未检测到棉酚,但从种子萌发12 h开始可以检测到较低含量的棉酚。种子萌发期色素腺体形成及棉酚合成相关基因表达谱分析结果表明,GoPGF基因在比克氏棉种子萌发12 h的子叶上表达量较高,与比克氏棉色素腺体形成有关;法尼基焦磷酸合成酶(FPS)基因在种子萌发24 h的子叶上表达量较高,可能与棉酚向子叶色素腺体转运有关;CYP706B1基因在种子萌发0 h较高,HMG1基因在种子萌发0 h、36 h和48 h的子叶上表达量均较高,说明这两个基因与种子萌发后色素腺体腔的形成有关,并参与棉酚的贮运;杜松烯合酶基因(CAD1-A)和HMG2基因与比克氏棉色素腺体形成无密切关系。【结论】种子萌发后的12 h为比克氏棉子叶色素腺体形态建成的关键时期,且色素腺体的形态建成与棉酚出现的时间具有同步性。GoPGF、FPS、CYP706B1、HMG1参与色素腺体的形成及棉酚的转运。     

比克氏棉和司笃克氏棉色素腺体形态建成的组织结构观察

对具有子叶色素腺体延缓形成性状的比克氏棉和种子有色素腺体但低酚的司笃克氏棉的休眠种子、幼胚发育过程和种子萌发过程中的子叶进行连续切片观察。结果表明,比克氏棉的种仁中虽无肉眼可见的色素腺体,但具有色素腺体原,其色素腺体形态建成从开花后子叶形态建成后开始到种子萌发后12h才完成。司笃克氏棉在幼胚发育过程中,也出现色素腺体原的结构,但保持的时间较短,到种子成熟时,其种仁中的色素腺体原大多已发育成正常的色素腺体,且色素腺体空腔中含有某些有色物质,腔壁明显不同于比克氏棉或一般有色素腺体棉。司笃克氏棉的色素腺体性状为介于一般有色素腺体棉和具有子叶色素腺体延缓形成性状棉种的中间过渡类型,这种性状在低酚棉育种研究中具有重要的实践意义。     

五个澳洲野生棉种色素腺体和棉酚性状研究

本文研究了斯特提棉（Ｇ．ｓｔｕｒｔｉａｎｕｍ）、南岱华棉（Ｇ．ｎａｎｄｅｗａｒｅｎｓｅ）、澳洲棉（Ｇ．ａｕｓ－ｔｒａｌｅ）、纳尔逊氏棉（Ｇ．ｎｅｌｓｏｎｉ）和比克氏棉（Ｇ．ｂｉｃｋｉ）五个澳洲野生棉种的种仁和植株各主要器官的色素腺体和棉酚及其旋光体含量。结果表明，五个棉种均具有种子无色素腺体、植株有色素腺体的特性，但五个棉种植株各器官上色素腺体的分布差异较大，特别是花瓣，其中斯特提棉和南岱华棉的花瓣具有正常的色素腺体，比克氏棉和澳洲棉的花瓣具有少量的色素腺体，纳尔逊氏棉的花瓣无色素腺体。五个棉种种仁棉酚含量极低，但植株各器官均含有较高的棉酚；棉酚旋光体含量测定结果表明，五个棉种的叶片中均只有左旋棉酚，而不含右旋棉酚，其中斯特提棉和南岱华棉两个种的左旋棉酚含量显著低于其余三个棉种     

棉属三染色体组(A、D、G)杂种的细胞遗传学研究

对棉属 A、D、G三个染色体组合成的三元杂种 [陆地棉× (亚洲棉×比克氏棉 ) ]及反交和回交后代 ,四元杂种 [(亚洲棉×比克氏棉 )× (陆地棉×夏威夷棉 ) ]进行了细胞学研究。结果表明 ,三元杂种 F1以及回交后代 BC4 F1分离出的不育株、四元杂种 F1不育的主要原因是花粉母细胞减数分裂异常 ,主要表现为 :大部分染色体为单价体 ,仅有少量配对成二价体 ,后期 两极分配不均匀 ,出现多极分离现象 ,并有落后染色体出现。末期出现大量多分孢子和不正常的四分孢子 ,从而不能形成正常的花粉粒。三元杂种回交后代 BC4 F1中的部分可育型 ,花粉母细胞进行减数分裂时 ,大部分染色体都能联会配对 ,仅个别为单价体 ,因而形成的花粉粒仅部分正常。 BC4 F1的可育型花粉母细胞进行减数分裂时 ,染色体配对正常 ,表明育性恢复 ,结实正常。此外 ,四元杂种花粉母细胞进行减数分裂时 ,在后期 形成了染色体“桥”     

(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。     

亚比棉×海岛棉杂种F_1的遗传分析

为了验证杂种的真实性,利用常规压片法和RAPD技术对亚比棉×海岛棉杂种F1及其亲本进行了研究。细胞学研究表明,3种杂种(亚洲棉×比克氏棉)×海岛棉的核型公式为:2n=4x=52=44m(2SAT)+8sm(2SAT),相对长度变异范围为6.91%～1.99%,臂比值为1.01～2.31之间,平均臂比为1.37,核型类型为2B。RAPD分析表明,亚比棉×海岛棉杂种F1共扩增出60条DNA带,其中具亚洲棉特异带的占总带数的3.3%;来自比克氏棉的带占8.3%;具海岛棉特异带占18.3%;来自亚比棉的特异带占10%;杂种的特异带占8.3%。     

亚洲棉、比克氏棉和海岛棉种间杂种的合成及性状研究

用（亚洲棉×比克氏棉）Ｆ１双二倍体为母本与海岛棉的不同色素腺体基因型进行杂交，获得（亚洲棉×比克氏棉）×海岛棉种间杂交三种杂种（简称（亚×比）×海三种杂种，下同）．（亚×比）×海三种杂种植株整体性状似海岛棉亲本，但营养生长旺盛，植株高大，并兼有三个亲本的一些形态特性。（亚×比）×海三种杂种花粉高度不育，但雌配子部分可育。海岛棉的不同色素腺体基因型对（亚×比）×海三种杂种的色素腺体在现有较大的影响，其中Ｇｌ２Ｇｌ２Ｇｌ３Ｇｌ３所产生的（亚×比）×海三种杂种，其种子尚有少量色素腺体，但植株为正常色素腺体类型；而ｇｌ２ｇｌ２ｇｌ３ｇｌ３所产生的（亚×比）×海三种杂种虽种子无色素腺体，但植株为少色素腺体类型，Ｇｌｅ２对（亚×比）×海三种杂种的色素腺体的影响与ｇｌ２ｇｌ２ｇｌ３ｇｌ３相同，只是杂种植株色素腺体的减少现象是更明显。此外，本文对比克氏棉子叶色素腺体延缓发生性状的遗传及其转育途径和技术进行了讨论。     

亚洲棉、比克氏棉和陆地棉异源四倍体的合成

为了将野生比克氏棉的种子无腺体,而植株上有腺体的珍贵特性转育到栽培棉种上,先用二倍体亚洲棉(A_2A_2)与比克氏棉(G_1G_1)杂交成异源二倍体A_2G_1,将杂种染色体加倍,合成异源四倍体 A_2A_2G_1G_1,再与栽培四倍体陆地棉杂交,组成三交种合成异源四倍体。并对三种杂种的性状遗传学及细胞学进行了分析研究。为培育棉、油、蛋白质三位一体的抗虫新棉种提供了种质。     

棉花海陆杂交纤维品质性状遗传参数分析

为了研究棉花海陆杂交纤维品质性状的遗传规律,本试验采用5个陆地棉品种（系）作母本与5个海岛棉品种（系）杂交,配制25个杂交组合,随机区组试验设计,重复2次。主要检测杂交种F1代纤维品质性状,以小区平均数进行相应的方差分析及遗传参数估计。结果表明：陆地棉亲本间变异除马克隆值外,其他纤维品质性状均差异显著或极显著,而海岛棉亲本间除伸长率外,其他性状均存在显著或极显著差异;各纤维品质性状双亲互作效应均不显著。另外,遗传参数结果表明：绒长、比强度和马克隆值的遗传变异系数与表型变异系数变化趋势基本吻合,且均大于环境变异系数。整齐度和伸长率的遗传变异系数均小于环境变异系数,说明整齐度和伸长率的变异中环境因素的作用不容忽视。绒长、比强度和马克隆值的广义遗传率较高,在采用海陆杂交改良棉花纤维品质性状时,适宜在早代选择优异株系。     

Relationship between molecular marker heterozygosity and hybrid performance in intra- and interspecific hybrids of cotton

The present study was conducted to investigate the relationship between parental molecular marker diversity and hybrid performance in both intra‐ and interspecific hybrids of cotton to evaluate the feasibility of predicting hybrid performance using molecular markers. Three cytoplasmic male sterile (CMS) lines were crossed with 10 restorer lines to produce 22 F₁ hybrids during 2003. Of 22 F₁s, 14 hybrids were intraspecific (Gossypium hirsutum × G. hirsutum) and eight interspecific (G. hirsutum × G. barbadense). These 22 F₁ hybrids and their parents were evaluated for yield and fibre quality traits at Zhejiang University, Hangzhou, China during 2004 and 2005. Genetic distances (GD) among the parents were calculated from 56 random‐amplified polymorphic DNAs (RAPD) and 66 simple sequence repeat (SSR) marker data, and their correlation with hybrid performance and heterosis were analysed. The parents could be discriminated into G. hirsutum and G. barbadense clusters by cluster analysis based on both RAPD and SSR markers data. The correlation (r = 0.503, P ≤ 0.05) was calculated between GD_rapd (GD based on RAPD markers) and GD_ssr (GD based on SSR markers). Correlation of GD with hybrid performance and heterosis differed considerably between intra‐ and interspecific hybrids. The correlation between GD and hybrid performance was non‐significant for most of traits within the hybrids of G. hirsutum species. However, it was significantly and positively correlated for fibre length, fibre strength and elongation in interspecific hybrids. The relationship between GD and heterosis was observed to be positively significant for boll weight within hybrids of G. hirsutum with significant and negative correlations for fibre length and elongation. In conclusion, the power of predicting hybrid performance using molecular markers in cotton is low. But, the relationship between SSR marker heterozygosity and hybrid performance can be used to predict fibre length during interspecific hybrid cotton breeding.     

High-performance liquid chromatographic analysis of anthocyanins in induced amphidiploids of Iris laevigata Fisch. × I. ensata Thunb.

Summary Anthocyanins of colchicine-induced amphidiploids of I. laevigata × I. ensata were analyzed by the HPLC procedure and compared with those of parental species and F₁ hybrids of I. laevigata × I. ensata. The amphidiploids were characterized by seventeen anthocyanins, and exhibited all anthocyanins of the parental species and the F₁ hybrids. Malvidin 3RGac5G and petunidin 3RGac5G in anthocyanins detected were major pigments in the amphidiploids, which exhibited strongly the malvidin 3RGac5G-petunidin 3RGac5G type of I. ensata rather than the petunidin 3RGac5G-malvidin 3RGac5G type of I. laevigata. Moreover, a higher degree of anthocyanin content was observed in the amphidiploids and the F₁ hybrids. Namely, the anthocyanin content of the amphidiploids and the F₁ hybrids were 2.81 and 2.45 times as much as that of the mid-parent. This implies that there is a complementary effect on anthocyanin synthesis of parental genes in them. Finally, the utility of the amphidiploids was discussed.Contribution from the Laboratory of Plant Breeding, Faculty of Agriculture, Miyazaki Univ., No. 64.     

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。     

Identification and Analysis of Fiber Quality and Yield Related Traits of Interspecific (Gossypium hirsutum L. × G. barbadense L.) Hybrids

[Objective] The aim of this study is to study the hereditary of heterosis of fiber quality and yield-related traits in the upland-island interspecific hybrids, and breed new interspecific hybrid varieties with high yield and fine fiber quality. [Method] In this study, 12 upland cotton materials and 5 sea-island cotton materials were selected to determine the fiber quality and yield traits of their parents and F₁ in Lin’an, Zhejiang and Sanya, Hainan. [Result] It was found that fiber length and fiber strength of F₁ (Gossypium hirsutum × G. barbadense) generally had significant mid-parent heterosis (MPH), some hybrid combinations showed strong over-parent heterosis (OPH), fiber length had a small coefficient of variation between the two places and could be stably inherited. And in terms of yield, seed cotton weight, lint weight, and lint percentage of some upland-island hybrids had MPH, but they were still significantly lower than those of upland cotton parents. [Conclusion] Two long-staple cotton hybrid combinations T035 and T044 with 5A grade high-quality were obtained, and an excellent material of G. barbadense Ta10-280 was screened. This study provides valuable data for the genetic law of fiber quality heterosis of upland-island hybrid cotton.     

哈克尼西棉细胞质对陆地棉主要经济性状的影响

利用2个恢复系与6个陆地棉品系正反交,结合人工辅助授粉,研究了杂种F1的经济性状表现。结果表明:杂种F1各经济性状广泛地受核基因控制;哈克尼西棉细胞质对F1的衣分、衣指、籽指及绒长有显著影响,且作用程度较高;多数性状不存在明显的核质互作效应。同核异质杂种相比较,哈克尼西棉细胞质F1衣指较低,籽指较高,衣分降低4~7个百分点;绒长增加1~2mm;纤维细度和断长也表现一定负效应。     

Somatic hybrids between Gossypium hirsutum L. (4×) and G. davidsonii Kellog (2×) produced by protoplast fusion

Somatic hybrids were obtained from electrofused protoplasts derived from embryogenic suspension cultures of tetraploid cotton (G. hirsutum L. cv. Coker 201) and embryogenic callus of diploid wild cotton G. davidsonii. The regenerants were initially identified as hybrids by RAPD (random amplified polymorphic DNA) analysis. Subsequently, observation on chromosome counting, morphology and SSR (simple sequence repeat) confirmed the hybrid status. Cytological investigation of the metaphase root-tip cells of the regenerated plants revealed there were 74 to 84 chromosomes in the plants, close to the expected 78 chromosomes. SSR analysis revealed the regenerated plants contained specific genomic fragments from both fusion partners, further confirmed their hybridity. The morphology of the plants was intermediate between the two fusion partners. The regenerants were difficult to develop into mature plants because their roots browned and they wilted from the stem apex before forming 3 to 5 true leaves. The hybrid plants were transferred to soil by grafting in vitro onto rootstocks.     

Application of Gibbereilic Acid on Diploid and Tetraploid Cotton Hybridization

Gibberellic acid growth regulator was used to develop interspecific hybrids between tetraploid and diploid species to increase the genetic variability in cotton.     

瑟伯氏棉和异常棉的陆地棉导入系的EST-SSR和gSSR分析

 用EST-SSR和gSSR分析了瑟伯氏棉和异常棉的8个导入系。结果表明,SSR的扩增带可以分为两大类：第一大类是导入系和受体晋棉6号之间没有差异;第二大类是导入系和受体晋棉6号之间存在差异。第一大类可分为5个小亚类;第二大类中,对于EST SSR来说,可分为4个小亚类：一是在晋棉6号中出现的条带在部分导入系中消失;二是条带大小与晋棉6号相比有变化;三是部分导入系中出现了瑟伯氏棉和异常棉的条带,但与晋棉6号相同的条带消失了;四是部分导入系中出现了供体和受体都没有的特异条带,而在另一些导入系中条带大小发生变化。对于gSSR来说,第二大类中又多了两个小亚类：即部分导入系中除了供体和晋棉6号都没有的条带;或供体和晋棉6号的条带同时出现在导入系中。这些供体和受体异常条带的增加和消失可能是SSR区域和SSR侧翼序列变化的结果。与gSSR相比,EST-SSR在导入系和晋棉6号之间的变化更大,这可能是导致导入系比晋棉6号纤维品质更优的原因。