脱落酸对棉花体细胞胚胎发生的影响

【目的】研究脱落酸(Abscisic acid, ABA)对棉花体细胞胚胎发生过程中下胚轴脱分化和再分化的影响,优化体细胞胚胎发生体系和初步解析脱落酸调控棉花体细胞胚胎发生分子机制。【方法】以棉花品种中棉所24(CCRI 24)下胚轴为外植体,设置5个ABA浓度0、0.02、0.04、0.06、0.08μmol·L^-1,分别以A0、A1、A2、A3、A4表示,添加至MSB(MS培养基+B5维生素)培养基诱导愈伤和胚性愈伤,研究ABA对棉花下胚轴初始细胞脱分化、愈伤组织诱导和胚性愈伤组织诱导的影响。【结果】ABA促进下胚轴初始细胞脱分化;显著提高愈伤组织的脱分化率和增殖率;0.02μmol·L^-1ABA显著提高胚性愈伤分化率,0.04~0.08μmol·L^-1ABA显著降低胚性愈伤分化率。ABA处理后胚性愈伤和非胚性愈伤的增殖率均显著提高且质地受到影响。0.02~0.08μmol ABA处理下,LBD和LBD在愈伤起始期上调表达。0.02μmol·L^-1ABA处理下,在愈伤增殖早期和中期BBM、LEC1和AGL15上调表达,愈伤增殖后期FUS3、LEA、ABI3基因上调表达。【结论】脱落酸调控的棉花体细胞胚胎发生与相关标记基因的时空性表达密切相关,这些基因表达水平的增加是ABA调控愈伤和胚性愈伤分化的分子基础。     

Construction of a genetic linkage map and QTL analysis of fiber-related traits in upland cotton (Gossypium hirsutum L.)

A genetic linkage map with 70 loci (55 SSR, 12 AFLP and 3 morphological loci) was constructed using 117 F₂ plants obtained from a cross between two upland cotton cultivars Yumian 1 and T586, which have relatively high levels of DNA marker polymorphism and differ remarkably in fiber-related traits. The linkage map comprised of 20 linkage groups, covering 525 cM with an average distance of 7.5 cM between two markers, or approximately 11.8% of the recombination length of the cotton genome. The present genetic linkage map was used to identify and map the quantitative trait loci (QTLs) affecting lint percentage and fiber quality traits in 117 F_2:3 family lines. Sixteen QTLs for lint percentage and fiber quality traits were identified in six linkage groups by multiple interval mapping: four QTLs for lint percentage, two QTLs for fiber 2.5% span length, three QTLs for fiber length uniformity, three QTLs for fiber strength, two QTLs for fiber elongation and two QTLs for micronaire reading. The QTL controlling fiber-related traits were mainly additive, and meanwhile including dominant and overdominant. Several QTLs affecting different fiber-related traits were detected within the same chromosome region, suggesting that genes controlling fiber traits may be linked or the result of pleiotropy.     

Genetic analysis of earliness in upland cotton ( Gossypium hirsutum L.). II. Yield and lint percentage

Inheritance and interrelationships of seed cotton and lint yields were evaluated in a diallel analysis involving seven early maturing parents of different origin and a commercial variety. Lint yield showed relatively little additive variance and low heritability, whereas lint percentage showed the opposite. Highest yields were shown by the least determinate and slowest-maturing genotypes; yields generally decreased as determinacy increased and rate of maturity accelerated. Except for date for first open boll, components of earliness showed no associated with yield.     

Genetic association of within‐boll yield components and boll morphological traits with fibre properties in upland cotton (Gossypium hirsutum L.)

Knowledge of genetic relationships between within‐boll yield components and fibre quality is essential for simultaneous improvement of lint yield and fibre quality in upland cotton (Gossypium hirsutum L.). Nine parents and their F₁ progeny with reciprocals from a 3 × 6 factorial mating design were grown in 2008 and 2009. Seven within‐boll yield components and two boll morphological traits and the three fibre quality parameters were analysed based on a conditional additive/dominance (AD) genetic model. Results showed that boll length contributed to the largest proportion of phenotypic, additive and dominance variances for UHM length; seed index contributed to the largest proportion of phenotypic and additive variances for fibre strength; boll width made the largest contribution to phenotypic and additive variances and the second largest contribution to dominance‐by‐environment interaction variance for micronaire, indicating that they played an important role than the other traits for fibre length, strength and fineness/maturity, respectively. It is worthy of note that those correlations between boll shape and fibre quality apply only to the nine parents and the resultant hybrids in this study and do not imply a cause and effect relationship.     

Lint development and properties of fifteen fuzzless seed lines of Upland cotton (Gossypium hirsutum L.)

Fifteen fuzzless seed lines in “obsolete” backgrounds of Upland cotton (Gossypium hirsutum L.) were obtained from the National Cotton Germplasm Collection and evaluated for fuzzless seed genotype, lint percent, and lint quality. Fourteen of these fifteen lines were found to be homozygous for the dominant fuzzless seed allele N ₁. Only one line was homozygous for the recessive fuzzless seed allele n ₂. The measured lint percent of each line was very stable through time, however, large variability existed between many of the N ₁ lines ranging from 0.7 to 23.6% lint. The lint percent for the n ₂ line was 24.4%. Scanning electron microscopy was used to differentiate patterns of lint initiation on 1 day post anthesis ovules. General patterns included: first, lint initiation restricted to the chalazal end of the seed crest; second, lint initiation along the seed crest and laterally around the chalazal end of the ovule; and third, lint initiation covered all but the micropylar end of the ovule. Lint quality was evaluated for each line using the Advanced Fiber Information System (AFIS) which included measurements of length, maturity, and fineness. The means of each measurement varied among the fuzzless seed lines with significant differences between fiber length, short fiber content, immature fiber content, fineness, and maturity ratio. No correlation was found between lint percent and any of the lint quality measurements. These lines will provide a valuable resource for the study of fiber initiation and lint quality.     

新疆陆地棉铃柄长与棉花产量及纤维品质的关系

摘要:【目的】为了解新疆棉花铃柄长与主要农艺性状、产量性状以及纤维品质之间的关联关系,【方法】通过两年比较试验,跟踪调查了棉花主栽品种（系）的主要农艺性状、产量性状以及纤维品质,总结了两年的主要气候数据,并将这些与铃柄长做了比较分析,【结果与结论】结果表明：棉花各品种的铃柄长与棉花主要生长阶段6-8月的最高平均气温呈负相关的关系;与平均最低气温和平均日照时数呈正相关;与有效果枝、单株铃、果枝始节和始节高度成正相关,与株高、无效果枝成负相关;与衣分、果枝数和折合亩产均呈正相关的关系,与单铃重呈负相关;与纤维的上半部平均长度、马克隆值呈正相关,与整齐度指数大致呈正相关,50%参试棉花品种（系）的铃柄长与纤维断裂比强度呈正相关的关系,剩余50%呈负相关的关系。     

新疆棉花4个主栽品种的体细胞胚胎发生及植株再生

以新疆4个主栽棉花品种新陆中20、新陆早24、新陆早33和03298为材料, 通过不同浓度的激素组合成功地诱导获得了体细胞胚并进一步发育成苗。研究发现, 所用的4种激素组合均能有效诱导愈伤组织, 其中又以0.02 mg L^-1或0.10 mg L^-1 KT和0.1 mg L^-1 2,4-D组合的诱导效果最佳; 两个诱导措施有利于胚性愈伤组织的产生, 即沿中柱纵切棉花下胚轴切段, 并以纵切面接触培养基; 愈伤组织诱导培养基中KNO₃用量加倍。挑选黄绿色、灰绿色或浅绿色的质地疏松的愈伤组织继代于无激素且KNO₃含量加倍的培养基中可产生胚性愈伤组织, 并在高比例KT/2, 4-D(0.05 mg L^-1或0.10 mg L^-1 KT和0.01 mg L^-1 2,4-D)促进下发育成胚。借助在培养基上垫滤纸产生干燥作用, 并间隔使用强透气效果的棉塞对培养三角瓶进行透气处理, 体细胞胚可成熟发育并产生根系发达的正常再生植株。应用此法, 4个实验材料在6~8个月内即可获得大量再生苗。     

光合作用信号途径调控陆地棉矮化的机理研究

【目的】了解光合作用信号途径基因调控陆地棉矮化的机理。【方法】以现蕾期陆地棉矮化突变体LA-1及其近等基因系LH-1茎尖为材料,通过同位素标记相对和绝对定量(Isobaric tags for relative and absolute quantitation, i TRAQ)结合液相质谱串联(LC-MS/MS)的定量蛋白质组学技术及定量反转录-聚合酶链式反应(Quantitative reverse-polymerase chain reaction, qRT-PCR)技术,分析两种材料中蛋白水平及m RNA水平基因表达情况,并通过生理生化方法检测两种材料光合能力差异。【结果】光合作用信号途径差异表达蛋白PsbO、PsaE、PsaH、PetF-1、PetF-2在LA-1中表达量下调,PetC和delta表达量上调。m RNA水平,除delta基因在两种材料中表达量无显著差别外,其它基因与蛋白水平具有同样的表达趋势。现蕾后,LA-1的净光合速率(P_n)、气孔导度(Cond)显著小于LH-1,蒸腾速率(Tr)极显著小于LH-1,而胞间CO2浓度(Ci)无显著差异。与LH-1相比,LA-1的实际光化学效率(Y_Ⅱ)、光系统Ⅱ(PS Ⅱ)的潜在活性(F_v/F_0)显著减少,叶绿素荧光非光化学猝灭(NPQ)显著增大。【结论】陆地棉矮化突变体LA-1的矮化与光合作用信号途径存在相关性,为进一步研究LA-1矮化的分子机理及棉花的矮化育种提供基础。     

三个陆地棉水孔蛋白基因的克隆与表达分析

从陆地棉SSH-cDNA文库的测序结果中得到一条具有完整ORF的陆地棉水孔蛋白基因序列,将其命名为GhAQP2。以该基因编码的氨基酸序列为探针,在棉花EST数据库经同源搜索得到2个相似性较高的EST,利用RACE技术获得其全长cDNA序列,将其基因命名为GhAQP3和GhAQP4。基因结构分析发现GhAQP2和GhAQP3各有4个外显子,3个内含子;GhAQP4有3个外显子,2个内含子。生物信息分析表明3个基因编码蛋白均含有6个跨膜区,2个NPA结构域,其氨基酸序列具备MIP超家族典型的蛋白保守区序列特征。多序列比对发现3个基因的氨基酸序列与其他物种PIP2类水孔蛋白氨基酸序列具有很高的同源性。qRT-PCR分析表明,GhAQP2在纤维伸长后期优势表达,GhAQP3在下胚轴和子叶中高表达,GhAQP4在纤维伸长前期优势表达,推测3个基因在不同的组织中发挥作用。GhAQP2在20DPA优势表达,为研究该基因在纤维伸长向次生壁加厚期转化过程中的表达调控提供了重要信息。     

Regulatory Mechanism of the Photosynthetic Pathway in Dwarfed Upland Cotton (Gossypium hirsutum L.)

[Objective] Here, we examined the mechanism of the photosynthetic pathway in dwarfed upland cotton (Gossypium hirsutum L.). [Methods] The upland cotton dwarf line LA-1 and the near-isogenic line LH-1 were used as research materials. We screened for dwarf-related differentially expressed genes at protein and mRNA levels by applying the isobaric tags for relative and absolute quantitation method in combination with LC-MS/MS quantitative proteomic technology and quantitative reverse-polymerase chain reaction. The difference in the photosynthetic abilities between the two materials were detected using physiological and biochemical methods. [Results] An analysis on the differential expression of proteins encoded by photosynthetic system elements revealed that PsbO, PsaE, PsaH, PetF-1 and PetF-2 were down-regulated, while PetC and delta were up-regulated in LA-1. The expression trends of the mRNA levels were the same as at the protein levels, except for those of the delta gene. The net photosynthetic rate, stomatal conductance and transpiration rate in LA-1 were lower than those LH-1, but the intercellular CO₂ concentration was not significantly different after budding. This indicated that the non-stomatal factor led to a decreased net photosynthetic rate in LA-1. Chlorophyll fluorescence detection revealed that the actual photosynthetic efficiency and potential photochemical activity of photosystem II in LA-1 were significantly decreased, while non-photochemical quenching was significantly increased. [Conclusion] The dwarfism of LA-1 is related to the photosynthetic pathway, and the results lay a foundation for exploring key dwarf-related genes and the molecular basis for dwarfism in upland cotton.     

南疆不同类型海岛棉品种棉铃内同化物积累与分配特性的研究

 以南疆主栽海岛棉品种为材料,分析不同花期铃部器官同化物质积累动态、再度分配及吐絮棉铃的分配率。结果表明：同化物积累随时间呈“S”型曲线,用Logistic方程进行拟合均达到显著水平。同化物积累达最大值所需时间表现为棉纤维＞种子＞铃壳,新海28比优系193提前5～10 d;同化物再度分配随着花期的推移,铃壳率不断增大,子重率、经济系数、子棉/壳重逐渐减小。铃壳对子棉增重的贡献率,新海28大于优系193;棉铃的胚珠数、种子数、结子效率随开花时间的推移逐步增大,新海28单铃子房胚珠数、单铃种子数比优系193略低,而结子效率稍高;新海28具有开花早,铃期短,铃壳率小,结子效率高,铃壳同化物质的再分配对子棉增重的贡献率、子重率、经济系数、子棉与干重的比值均大于优系193,这是其早开花、早结铃、早吐絮及丰产的遗传生理基础。     

陆地棉钾转运体基因GhHAK5启动子的克隆与功能分析

KUP/HAK/KT钾转运体基因的转录调控是植物响应低钾胁迫的一项重要机制。克隆和分析棉花钾转运体基因的启动子,不仅有助于了解其表达模式及调控机制,对于改良棉花的钾吸收特性也具有重要意义。陆地棉钾转运体基因GhHAK5是一个在根中特异性高表达的基因,其表达受低钾胁迫诱导,目前关于该基因启动子的功能还不清楚。本研究以陆地棉品种百棉1号为材料,通过PCR方法对GhHAK5上游2000bp启动子片段(pGhHAK5)进行克隆,并通过转化拟南芥、GUS组织定位和低钾诱导表达特性分析来研究其功能。结果表明, pGhHAK5除具有TATA-box和CAAT-box等基本顺式作用元件外,还含有多个响应于光、逆境胁迫、植物激素和生物钟等的顺式作用元件。pGhHAK5与雷蒙德氏棉pGrHAK5在重要调控元件的数量和位置分布上具有较高的一致性,均具有5个参与根特异性表达调控的元件(ATAAAAT)和1个参与低钾条件下转录调控的ARF转录因子结合位点(TGTCNN)。GUS组织化学染色结果显示,转基因拟南芥幼苗的叶脉和胚轴维管束组织染色较深,根系染色较浅;成熟期转基因拟南芥植株的根、叶脉和花萼维管束组织染色...     

Association mapping identifies markers related to major early‐maturating traits in upland cotton (Gossypium hirsutum L.)

Association mapping is a promising tool to identify genes for quantitative traits. A total of 172 upland cotton (Gossypium hirsutum L.) accessions were assessed. The panel was phenotyped for four major early‐maturating traits across multiple environments and genotyped using 331 polymorphic simple sequence repeat (SSR) markers. The gene diversity of the 331 markers averaged 0.40 (range: 0.04–0.78), and the polymorphism information content (PIC) averaged 0.34 (range: 0.04–0.75). Thirty‐nine significant markers related to early‐maturating traits were simultaneously detected in at least two environments. Among the 39 markers, 11 for seed period (SP), seven for bud period (BP), six for flower and boll period (FBP), and 15 for growth period (GP). These markers were stable and could be used for marker‐assisted selection (MAS). Favourable alleles for SP, BP, FBP and GP were explored. These alleles can be directly used in MAS to improve cotton early maturity. This study lays the foundation for analysing the genetic mechanisms underlying early maturity, as well as the use of MAS to target traits in cotton.     

陆地棉产量组分对皮棉产量的遗传贡献分析

采用加性-显性-母性及其与环境互作的遗传模型,对5个陆地棉亲本及其F1代20个组合6个产量组分和皮棉产量的两年资料,利用估算条件方差分量和预测条件遗传效应值的统计方法进行了贡献分析.结果表明,产量组分对皮棉产量表型值的贡献变化范围为-46%～80%;铃数对皮棉产量有很高的显性贡献率(CRD=96%),其次是铃重(CRD=41%);铃数对皮棉产量的母性×环境互作遗传方差贡献率最大(CRME=98%),其次是铃重(CRME=77%)、衣指(CRME=62%)和子指(CRME=42%);铃数对皮棉产量的加性、母性效应贡献因亲本而异;对亲本皮棉产量的母性×环境互作贡献最大的产量组分性状因不同亲本而有所不同,表明各亲本具有其独特的遗传和发育特性,亲本1的铃重对其皮棉产量的母性效应在环境1中有最大的贡献(MEi(C→T)= 5.49 g),在环境2中,亲本3的铃重有最大的母性×环境互作效应贡献(MEi(C→T)=0.71 g),亲本5的铃数具有最大的母性×环境互作效应贡献(MEi(C→T)=0.51 g).多数杂交组合皮棉产量的显性效应主要受铃数和铃重影响,因此铃数和铃重可作为间接选择组合皮棉产量显性效应的指标.     

The inheritance model for the fiberless trait in upland cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) line SL1-7-1: variation on a theme

Segregating populations were developed to evaluate the inheritance of the fiberless seed phenotype of upland cotton (Gossypium hirsutum L.) line SL1-7-1. We report the inheritance of fuzzy, fuzzless and fiberless seed from crosses of SL1-7-1 with wildtype DP5690, Mexican fuzzless seed UA 3-3 (accession 143), Ballard fuzzless seed (accession 243), and MD17. Results from the F₁, F₂ and F_2:3 progeny derived from the SL1-7-1 X DP5690 indicated that the expression of the fiberless phenotype fit a three loci model with one locus being the dominant fuzzless seed allele N ₁ . The other two loci were tested to verify whether they were allelic to either recessive fuzzless seed alleles n ₂ or n ₃ . Using the segregation ratios of the F₂ progeny derived from the 143 X SL1-7-1 cross and F₂-derived F₃ families from SL1-7-1 X DP5690 with fuzzy seed (lacked N ₁ ), it is proposed that SL1-7-1 lacks the recessive n ₂ allele, but contains the n ₃ allele in the genotype of SL1-7-1. The third locus was previously not characterized and has been designated as fl ₁ (fiberless), therefore, the genotype for the fiberless phenotype of SL1-7-1 is N ₁ N ₁ fl ₁ fl ₁ n ₃ n ₃ . Fiberless lines MD17 X SL1-7-1 were crossed to verify similarities in genotypes between line and the genotype model predictability. Various combinations of the homozygous and heterozygous expression of N ₁ , n ₂ , n ₃ and fl ₁ allele produced plants with lower lint percentages.     

Hydrogen peroxide reduces heat‐induced yield losses in cotton (Gossypium hirsutum L.) by protecting cellular membrane damage

We investigated the effect of various growth substances such as hydrogen peroxide, salicylic acid (SA), moringa leaf‐extract (MLE) and ascorbic acid (ASA) on leaf physiology and seed cotton yield (SCY) of heat‐stressed cotton. Cotton plants were exposed to elevated temperatures at three reproductive stages, either by staggering planting time in the field or by increasing growth cabinet temperatures (38/24°C and 45/30°C) in glasshouse. Elevated temperature at any reproductive phase significantly damaged cellular membrane and reduced SCY. Plants exposed to 38/24°C and 45/30°C in glasshouse produced 63% and 22% lower SCY, respectively, compared with plants under optimal temperature ((32/20°C). In response to high temperature, cotton plants up‐regulated activities of anti‐oxidative enzymes e.g. peroxidase and ascorbic acid. However, this defensive system could not protect cellular membrane of stressed plants from extreme temperature (38 and 45°C). In contrast, growth substances such as H2O2, ASA and MLE significantly increased anti‐oxidative enzymes activity to an extent, which reduced heat‐induced damage to cellular membrane. No significant effect of any regulator was observed on SCY under optimum temperatures; although H2O2, MLE and ASA significantly increased SCY of heat‐stressed cotton. Hydrogen peroxide increased SCY of April and May thermal regimes crops by 16% (averaged across both sowing dates) under field, while it caused 14% and 20% increase in SCY of plants exposed to sub (38/24°C) and supra optimal (45/30°C) thermal regimes under glasshouse. We concluded that growth regulators, specifically, H2O2 can protect cotton crops from heat‐induced cellular membrane damage by up‐regulating antioxidant defense system.     

棉花磷胁迫响应基因GhWRKY6克隆及功能分析

【目的】提高棉花在低磷胁迫下的抗性,挖掘棉花磷胁迫相关功能基因。【方法】用生物信息学方法分析Gh WRKY6基因的结构特征和进化关系;构建基因超表达载体转化拟南芥,分析转基因拟南芥在低磷胁迫下的抗性;利用荧光定量PCR(Real-time quantitative polymerase chain reaction, qRT-PCR)分析拟南芥中磷信号途径Marker基因表达量的变化。【结果】以陆地棉cDNA为模板克隆得到Gh WRKY6基因,生物信息学分析表明该基因序列含有一个WRKY保守结构域,是WRKY家族转录因子。在低磷胁迫下,转基因拟南芥与野生型相比,种子萌发速度、主根长度、侧根数目、生物量均低于野生型,而在正常生长条件下两者并无差别。qRT-PCR分析表明GhWRKY6能够影响关键的磷转运蛋白基因的表达。【结论】GhWRKY6作为一个负调控因子参与到植物低磷胁迫响应信号途径中。