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。     

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.     

澳洲野生棉种的子叶棉酚动态及与色素腺体形态的关系

对斯特提棉、南岱华棉、澳洲棉、纳尔逊氏棉和比克氏棉五个澳洲野生棉种在种子萌发过程中的子叶棉酚含量进行了测定和分析。结果表明,五个澳洲野生棉种子叶中的棉酚积累过程与无色素腺体陆地棉品种基本相似,其休眠种子和萌发２４ｈ以内的子叶均检测不到棉酚,为典型的低酚棉类型。当种子萌发４８ｈ以后,其子叶开始出现棉酚,但含量极低;在子叶开始出现棉酚１～２ｄ后,棉酚含量开始迅速增加,到种子萌发第９ｄ达到最大值。此外,讨论了澳洲野生棉种的子叶色素腺体延缓形成的组织结构与棉酚积累的关系,以及该性状的遗传调控机理等。     

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.     

一个新的棉花腺体形成基因的遗传鉴定

湘X9628陆地棉突变体具有植株有腺体、种子低棉酚的特性。种子棉仁中的棉酚含量为0.0354%,低于国家标准。它的种子无腺体,当种子萌发后,长出的真叶有腺体,但比常规陆地棉品种少。遗传分析表明,控制湘X9628植株有腺体、种子低棉酚特性由二对重叠隐性基因所控制。等位性测验表明,其中一对基因是gl2的等位基因,另一对是gl     

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。     

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.     

澳洲野生棉种子叶色素腺体延缓形成的组织结构观察

对５个澳洲野生棉种休眠种子、种子发育过程中的子叶和种子萌发过程中的子叶进行连续切片观察,并以比克氏棉为例进行透视电镜观察。结果表明,具有子叶色素腺体延缓形成性状的澳洲野生棉种及其杂交后代的休眠种子,虽肉眼看不到色素腺体,但具有一群不同于其它细胞的特殊细胞群,其整体结构类似色素腺体的雏形。因此,提出了“色素腺体原”的新概念。澳洲野生棉种在开花后的幼胚器官形态建成期均先出现色素腺体细胞,然后经数次分裂形成一团色素腺体细胞群,挤压周围细胞而形成为色素腺体原,并保持该结构至种子成熟。当种子萌发１６ｈ后,色素腺体细胞解体,形成色素腺体。此外,本研究对澳洲野生棉种子叶色素腺体延缓形成性状的遗传调控等问题进行了讨论。     

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。     

棉酚合成及棉花腺体形成相关基因的研究进展

棉酚是限制棉籽开发利用的重要因子。培育低酚棉,特别是采用基因工程方法创制低酚棉是解决棉籽开发利用难题的关键。棉酚合成以及棉酚储存场所棉花腺体形成相关基因的研究,是基因工程创制低酚棉的基础。综述了国内外对于棉酚合成途径、棉花腺体形成相关基因的研究进展以及利用基因工程手段创制低酚棉的相关实例,以期为相关研究者提供参考。     

Genetic variation and selection within glandless cotton germplasm

There is a renewed interest in research and use of glandless (free of gossypol) Upland cotton (Gossypium hirsutum L.) that can produce edible seeds for human food and animal feed. However, there was a lack of information on yield potential of existing glandless germplasm since intermittent breeding activities for glandless cotton were stopped in the U.S. before 2000. The objectives of the present study were to test obsolete and exotic glandless germplasm for possible production and evaluate progress in direct pedigree selection within the existing glandless germplasm in eight field tests. In multiple tests, the glandless cotton Acala GLS yielded only 65–80 % lint of the glanded control Acala 1517-08 and 46–75 % lint of commercial transgenic cultivars. Most of obsolete glandless germplasm and their selections yielded <70 % of Acala 1517-08 and only three selections yielded 82–89 % of the control. Genetic variation in yield and fiber quality traits was seen from significant differences between selections within the same glandless germplasm, indicating the existence of residual genetic variation. Most importantly, three selections from exotic glandless germplasm out-yielded Acala 1517-08 by 4–9 %, one of which had desirable fiber quality traits. These promising glandless lines with comparable yields to commercial cotton make it possible to revitalize the glandless cotton industry. More field tests will be needed to evaluate their yield potential and adaptability in the U.S. Cotton Belt.     

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。     

司笃克氏棉（G.stockii）色素腺体的形态建成与棉酚动态研究

对司笃克氏棉幼胚发育过程和种子萌发过程中的色素腺体形态建成进行连续的切片观察和棉酚含量分析。结果表明,司笃克氏棉的种仁中含有肉眼可见的色素腺体,为典型的有色素腺体类型;然而种仁中的棉酚含量仅为0.0058%,为典型的低酚棉类型。司笃克氏棉种子成熟过程中的色素腺体形态建成有两种情况：（1）色素腺体细胞直接形成     

色素腺体和棉酚对陆地棉茎尖培养的影响

本研究以陆地棉三对色素腺体近等基因系为材料,研究色素腺体和棉酚对棉花茎尖培养的影响.结果表明,外植体的色素腺体和棉酚性状对于棉花茎尖培养有较大的影响,有色素腺体棉的茎尖比对应的无色素腺体近等基因系更易诱导产生不定根和再生植株.无色素腺体棉根尖的不定根出生时间晚于对应的有色素腺体近等基因系7天左右.棉酚试验     

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。     

Expression of the “<Emphasis Type="Italic">glanded-plant</Emphasis> and <Emphasis Type="Italic">glandless-seed</Emphasis>” trait of Australian diploid cottons in different genetic backgrounds

The expression of the “glanded-plant and glandless-seed” trait was assessed using High Performance Liquid Chromatography (HPLC) analysis methods in different Gossypium hybrids obtained by crossing Australian diploid cottons and various diploid and tetraploid species. Significant variation in the gossypol content in the seed was observed among the analyzed genotypes. HPLC data demonstrated that the gossypol synthesis repression mechanism in the Australian diploid species belonging to C and G genomes was dominant but did not confirm its preferential functioning against A genome species bearing GL₂ locus. About 10% of the produced seeds had total gossypol content lower than the limit imposed by the World Health Organisation (600 ppm) for the use of cotton flour in food and feed. HPLC analysis of the terpenoids aldehyde (TA) contents in the aerial parts of the hybrids showed important qualitative and quantitative variability. This result could indicate a certain separation between pigment gland morphogenesis and terpenoid synthesis mechanisms in cotton.     

Lowering seed gossypol content in glanded cotton (Gossypium hirsutum L.) lines

Cottonseed is a rich source of high quality protein, but its value as an animal feed is limited by gossypol, a toxic polyphenolic compound contained in glands located throughout the plant. This compound helps protect the plant from pests. Totally glandless varieties have been developed, but not adopted as these plants are left vulnerable to pests. This study describes a breeding strategy to decrease the levels of gossypol in the seed while maintaining a high enough concentration of toxin in vegetative plant parts to offer protection from pests. Preliminary studies indicated that crosses between varieties with different gland densities and distributions produced a range of glanding patterns. By selecting within the resulting progeny, we have identified F₇ generation progeny that have <0.30% total gossypol in the seed, while still possessing glands at critical locations on the vegetative plant parts. These new lines will be a valuable source of germplasm for developing low seed gossypol varieties. Seed from these varieties would provide a new source of inexpensive protein for animal feeding rations.