棉花幼苗和离体根无性系中棉酚的动态变化研究

以陆地棉三对色素腺体近等基因系以及海岛棉不同色素腺体基因型为材料 ,对种子萌发后的子叶、根系和离体根系无性系的棉酚及其旋光体含量进行了系统的分析 ,结果表明 ,有色素腺体棉种子萌发后的子叶棉酚含量呈下降趋势 ,而无色素腺体棉则呈积累趋势 ;棉花根系中的棉酚含量与子叶有相同趋势 ,但变幅显著小于子叶。有色素腺体棉和无色素腺体棉的离体根系无性系中的棉酚含量均随着培养时间而持续增加 ,培养 2 5d的棉花根系无性系中的棉酚含量可达到 0 .8%～0 .9% ,说明棉酚主要由棉花根系合成。棉酚旋光体含量测定结果表明 ,棉花种子萌发初期的子叶左旋棉酚和右旋棉酚含量与其种仁相一致 ,但种子萌发 1 4d后 ,二者的含量几乎相等 ;离体根系无性系 ,以及无色素腺体棉的子叶和根系中的棉酚均为消旋棉酚 ,表明棉花生长过程中合成的棉酚为消旋棉酚。此外 ,本文还就棉酚及其旋光体的合成和代谢进行了讨论。     

棉花色素腺体和棉酚对棉铃虫抗药性的诱导作用及其酯酶同工酶谱分析

本研究以3对陆地棉色素腺体近等基因系为材料,结合不同棉酚含量的半人工饲料,研究棉花色素腺体和棉酚对棉铃虫抗药性的诱导作用。结果表明,取食于有色素腺体棉叶的棉铃虫,其抗药性显著高于取食于相应的无色素腺体近等基因系棉叶的棉铃虫,色素腺体可诱导棉铃虫对菊酯类和有机磷类农药产生抗药性。不同棉酚含量的半人工饲料试验结果表明,在一定范围内,棉铃虫的抗药性随饲料中的棉酚含量的增加而增加,当棉酚含量超过0．3％时,初孵棉铃虫难于饲养成活。棉铃虫体内酯酶同工酶谱分析结果表明,色素腺体对棉铃虫体内酯酶同工酶谱的谱带数无显著影响,但对其中一些谱带的宽度和颜色有一定程度的影响;棉酚的作用与色素腺体相似,但饲以0．225％的棉酚的棉铃虫体内出现两条新的谱带。经鉴定,新出现的谱带为羧酸酯酶。     

棉花黄萎病抗性与过氧化物酶同工酶分析

采用等电聚焦电泳方法 ,对棉花二对抗黄萎病近等基因系以及三对色素腺体近等基因系的子叶进行过氧化物同工酶 (POD)分析。结果表明 ,色素腺体和棉酚含量与棉花抗性没有必然的联系 ;环境条件对棉花抗性表现有一定的影响 ,但对子叶中 POD同工酶谱带无显著的影响 ;4条 POD同工酶酶带 (P4、P6、P7、P10 )的表现与棉花抗黄萎病性有关 ,这些酶带的特殊表现可用于棉花抗病性辅助鉴定和辅助育种     

棉花无腺体近等基因系差异表达基因分析

为了揭示棉花腺体发育的分子机制,本研究利用RNA-Seq对棉花无腺体性状近等基因系Z12/Z12YW的幼嫩叶片进行转录组测序,寻找差异表达基因。转录组测序结果表明,与有腺体棉Z12相比,无腺体棉Z12YW中共有306个基因表达发生变化,其中282个基因下调,24个基因上调。差异表达基因GO(Gene ontology)功能注释显示,细胞组分、胞外基质组分的功能变化以及细胞杀伤生物学过程在腺体形成过程中发挥重要作用;COG(Cluster of orthologous groups)功能和KEGG(Kyoto encyclopedia of genes and genomes)通路显著性富集发现,棉酚等萜烯物质的合成主要受甲羟戊酸途径上游基因的表达调控。此外,本研究共筛选到13个差异表达转录因子,可能在腺体发育和棉酚合成过程中发挥关键作用。     

无棉毒素棉花品种的引进和利用

目前生产上栽培的棉花在棉籽仁和根、茎、叶等部位的色素腺中,含有黑褐色的腺体,俗称油腺。这种腺体是一种多酚类的物质,其中以棉酚含量最高,占24.4—30.4%,其他还有许多棉酚的衍生物质,它们与蛋白     

棉花腺体性状基因的遗传与克隆研究进展

摘要：棉酚腺体是棉属植物特有的性状,发掘和利用控制腺体和棉酚合成的基因,对深入阐明棉酚的生物合成机理以及通过基因工程手段培育新型棉花品种具有重要的理论和现实意义。本文综述了近年来棉花棉酚腺体性状基因的遗传与克隆研究的最新进展,并对该研究中存在的问题进行分析和展望。     

陆地棉不同色素腺体基因对比克氏棉子叶色素腺体延缓形成性状的遗传效应

用(亚洲棉×比克氏棉)F1双二倍体作母本与陆地棉的不同色素腺体基因型进行杂交,获得5个具有不同色素腺体基因的［(亚洲棉×比克氏棉)F1双二倍体×陆地棉］F1种间三元杂种。不同色素腺体基因对种间三元杂种植株和花器形态性状无明显效应,但对其色素腺体表现和棉酚含量具有较大的影响。用陆地棉单节显性系1(G12G12gl3gl3)配制     

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。     

棉酚旋光体柱前衍生化-HPLC分析与4个栽培棉种之间棉酚含量比较

采用2%D-氨基丙醇+10%冰醋酸+88%乙腈作为棉酚提取和衍生化试剂对消旋棉酚和棉花样品进行热衍生化处理,用C18色谱柱和乙腈流动相进行高效液相色谱分析。该方法色谱结果表现为主峰清晰而杂峰少,左右旋棉酚出峰时间相差7 min,实验精度良好。用该方法对4个栽培种的25份材料的种仁和幼苗的根、茎、叶的左右旋棉酚含量进行了系统的检测。结果表明,海岛棉的种子和根系的总棉酚含量最高,其次是陆地棉;2个二倍体栽培棉种的种子和根系的总棉酚显著低于四倍体棉种,而2个二倍体栽培棉种的叶片总棉酚含量显著高于四倍体棉种。大多参试材料的种子和幼苗各部位的左右旋棉酚含量均有差异,其中海岛棉的左旋棉酚含量略高于右旋棉酚,陆地棉的右旋棉酚含量略高于左旋棉酚,而2个二倍体棉种的左旋棉酚只占总棉酚的1/3。无色素腺体棉的种仁和叶片中总棉酚含量极低,并以左旋棉酚为主,其种仁中只有左旋棉酚而无右旋棉酚;茎秆中总棉酚含量以及左右旋棉酚比例与一般有色素腺体常规棉相似,但根系中的总棉酚含量显著高于对应的有色素腺体棉,却以右旋棉酚为主。     

陆地棉色素腺体与不同棉酚旋光体含量之间的相关性研究

【目的】研究不同基因型棉花不同器官的色素腺体与棉酚旋光体含量之间的相关性。【方法】以6个陆地棉材料中棉所12、中棉所12(无)、珂字棉312、珂字棉312(无)、IF11和冀棉14为对象,精确测定主要器官的色素腺体密度和大小,并采用柱前衍生拆旋方法进行高效液相色谱分析,测定其相应的棉酚旋光体含量,然后对色素腺体和棉酚旋光体含量进行相关性分析。【结果】低酚棉全株无色素腺体,但在各器官中仍能检测到低于0.02%(质量分数)的少量游离棉酚存在。有酚棉各器官均具有典型的色素腺体,其中棉仁的色素腺体最密,苞叶最稀;色素腺体大小(直径)以铃壳最大,花瓣和棉仁最小。不同器官中,总棉酚含量以棉仁最高,苞叶最小;棉仁和花瓣的右旋棉酚含量大于左旋棉酚,而萼片、铃壳、上部叶、中部叶和下部叶的左旋棉酚含量大于右旋棉酚。有酚棉的色素腺体密度与大小之间呈极显著负相关;而总棉酚含量与不同棉酚旋光体含量之间,以及左右旋棉酚含量之间呈极显著正相关。色素腺体密度与总棉酚含量以及不同棉酚旋光体含量之间呈极显著的正相关,而色素腺体大小与右旋棉酚含量之间呈显著的负相关。【结论】棉仁与花瓣的总棉酚含量存在高度的相关性,因此,可以通过花瓣的棉酚含量来判断棉仁的棉酚含量。     

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。     

陆地棉植株上色素腺体空间分布模式的多样性及其选择

利用陆地棉全株有色素腺体与无色素腺体的品种杂交而获得的6世代群体,研究了植株13个器官色素腺体的分布.基于器官上色素腺体的有无,发现植株色素腺体的空间分布具有10种模式.每一模式代表着植株色素腺体密度的特定等级,不同模式所包含的着生色素腺体器官的种类和数量不同.同一模式中各器官的色素腺体密度与其群体平均密度值的差亦不同,据此,作者提出了色素腺体的早生器官、迟生器官、同生器官和缓生器官的概念,并运用空间分布模式,为在实践中选样或保留理想模式提出了更为可靠的方法.     

棉酚及色素腺体对棉花抗枯萎病性的作用

本文对低酚棉与有酚棉的抗病和感病品种及其杂交后代进行了研究。结果表明，在种子中棉酚含量及色素腺体密度与品种的抗枯萎病性没有相关性。在幼苗中，低酚棉幼苗棉酚含量均显著高于有酚棉品种，但它们之间的抗枯萎病性也没有显著差异。在低酚棉与有酚棉品种杂交的Ｆ２代群体中，无色素腺体和稀色素腺体类型苗的抗病性明显高于腺体密度较高的类型苗。对７个杂交组合Ｆ２代中有色素腺体和无色素腺体幼苗的抗病性分析表明，无色素腺体类型苗的病指显著低于有色素腺体类型苗的病指。幼苗人工接种枯萎菌后，棉酚含量表现升高，但抗病品种低于感病品种。     

Effects of the dominant glandless gene Gl2e on agronomic and fibre characters of Upland cotton

Seven pairs of near‐isogenic lines (glandless vs. glanded) and the recurrent parents were used to determine the effects of the dominant glandless gene from ‘Hai 1’(Gossypium barbadense) on agronomic, fibre, and seed characters in Upland cotton, Gossypium hirsutum, backgrounds. The results showed that there were no apparent linkage associations of the glandless gene on most agronomic, fibre and seed characters of Upland cotton, except for seed quality. The glandless line derived from ‘Liaomian 7’had significantly more protein (489.6 g/kg), and that from H237 had significantly more oil (362.4 g/kg) and had the largest oil index (2.70 g) and protein index (3.03 g). The gossypol content of seed in dominant glandless lines in Upland cotton was very low (<0.04 g/kg). Therefore, it is suggested that the glandless gene can play an important role in breeding glandless or low seed‐gossypol Upland cotton cultivars.     

Genetic variation and comparative analysis of thrips resistance in glandless and glanded cotton under field conditions

Glandless cotton can be grown for cottonseed free of toxic gossypol to be used as food and feed for non-ruminant animals. However, one of the most important limiting factors preventing its commercial production is its higher insect damage than conventional glanded cotton. Thrips is the one of the most important insect pests in the early growing season that may cause yield losses. In this study, 28, 29, 26, and 2 glandless cotton lines were compared with glanded control Acala 1517-08 and other glanded lines for resistance to the Western flower thrips, Frankliniella occidentalis (Pergande) in four replicated field tests each containing 32 genotypes. In the same field, 28 glanded commercial cultivars and 78 glanded breeding lines were compared with Acala 1517-08 and Acala 1517-99 in three other tests with 32 genotypes each. The experimental layouts allowed a comprehensive comparative analysis of thrips resistance within and between glandless and glanded cotton. Overall, glandless cotton had similar or lower thrips damages than glanded cotton, indicating that the glandless trait may serve as a genetic factor for suppressing thrips damage. As compared with Acala 1517-08 which represented one of the most thrips resistant genotypes among glanded cotton tested, glandless Acala GLS and many selections from glandless germplasm were more resistant, while some were similar to Acala 1517-08, indicating that genetic factors other than the glandless trait also affect thrips resistance in cotton. The estimates for broad-sense heritability for thrips resistance were moderate, indicating that thrips resistance is selectable. This is corroborated by the identification of many thrips resistant lines from a cross between Acala 1517-08 and Acala GLS. This study has laid a foundation for a more detailed study using most resistant lines with desirable agronomic traits in multiple environments.     

Introgression of a gene for delayed pigment gland morphogenesis from Gossypium bickii into upland cotton

The presence of gossypol and its derivatives above the WHO/FAO standards (0.02–0.04%) in cotton seed oil and meal limits its usage as food and feed. To the contrary, the presence of pigment glands filled with gossypol and its derivatives helps to protect cotton plants from phytophageous pests. Thus a desirable cultivar would have glandless seeds on a glanded plant. This paper describes results on the successful introgression of this trait from Gossypium bickii into cultivated upland cotton. Five different tri‐specific hybrids (ABH1, ABH2, ABH3, ABH4 and ABH5) were obtained by crossing the amphidiploid F₁ (G. arboreum × G. bickii) with different gland genotypes of G. hirsutum as male parent. The hybrids were highly sterile, and their chromosome configuration at meiosis metaphase 1 (M1) in pollen mother cell (PMC) was 2n = 52 = 41.04 I + 4.54 II + 0.57 III + 0.04 IV. All five hybrids were similar in morphological characters, except for the gland expression and gossypol contents. The hybrid (ABH3) derived from genotype Gl₂Gl₂gl₃gl₃ of upland cotton (a single gene dominant line) had completely introgressed the target trait of G. bickii. While ABH1 and ABH2, which derived from recessive (gl₂gl₂gl₃gl₃) or dominant (GlGl) glandless upland cotton genotypes, had glandless seeds too, but the density and size of the glands on the plant were reduced significantly.     

Development of high-gossypol cotton plants with low-gossypol seeds using trispecies bridge crosses and in vitro culture of seed embryos

The objective of this work is to develop an upland cotton, Gossypium hirsutum L. [2n = 4x =52, 2(AD)h], having a reduced level of gossypol in the seeds for food and feed uses, and a high level of gossypol in the remaining organs to limit pest incidence. Using G. sturtianum Willis (2n = 2x = 26, 2C1) as donor and G. thurberi Torado (2n = 2x = 26, 2D1) or G. raimondii Ulbrich (2n = 2x = 26, 2D5) as bridge species, two trispecies hybrids G. thurberi– G. sturtianum– G. hirsutum and G. hirsutum– G. raimondii– G. sturtianum were synthesized. Both trispecies hybrids were male sterile. Recurrent backcrossing to G. hirsutum as pollinator and selfing of the second backcross (BC) progenies resulted in seeds which were rescued by in vitro culture. In total, 1208 flowers of the trispecies hybrids and their BC progenies yielded 192 seed embryos from which 62 plants were obtained. Cytogenetic analyses indicated a relatively high frequency of chromosome pairing and chiasmata. The gland levels in backcross seeds ranged from glandless seeds to normally glanded seeds. All vegetative parts of those hybrids were glanded, but a wide range of variability for gland density was observed on leaf, stem, bract and calyx. Plants derived from seeds having a reduced level of gossypol constitute very interesting germplasm to develop a cultivated glanded cotton with low-gossypol seeds. This revised version was published online in July 2006 with corrections to the Cover Date.