低酚棉种仁含油量的母体效应和杂种优势分析

利用3个低酚棉品种配制的四个正、反交组合的F1、F2和回交世代对棉籽种仁含油量的母体效应和杂种优势进行了遗传分析。结果表明，棉籽的种仁含油量受母体基因型的控制，即存在母体效应，细胞质效应对种仁含油量无显著影响。用种仁含油量高的品种作母本配制的杂种一代（F1）棉籽，其种仁含油量有极显著的中亲和高亲优势，F2棉籽种仁含油量的自交衰退率虽较大，但其中亲优势仍较大，且优于用低种仁含油量品种作母本配制的杂种二代（F2）棉籽。利用杂种优势是提高棉籽种仁含油量的有效方法，其中的关键是亲本选配。此外，本文还讨论了棉籽含油量遗传研究的模型选择问题和提高棉籽含油量的育种方法问题。     

Identification of resistance gene analogs in cotton (Gossypium hirsutum L.)

Sequence analyses of numerous plant disease resistance genes have revealed the presence of conserved motifs common to this class of genes, namely a nucleotide binding site (NBS) and leucine rich repeat region. In this study, thirty-three resistance gene analogs (RGAs) were cloned and sequenced from cotton (Gossypium hirsutum L.) following PCR with degenerate primers designed from the conserved NBS motif of plant resistance (R) genes. Phylogenetic analysis of the predicted amino acid sequences grouped the RGAs into four distinct classes from which several subgroups were delineated based on nucleic acid sequences. Gene database searches with the consensus protein sequences of each of the four classes and respective subgroups of cotton RGAs revealed their conserved NBS domains and homology to RGAs and known resistance genes from a variety of plant genera. Given the complete lack of knowledge regarding molecular organization of R genes in cotton, the cloned RGAs described here may be useful as probes to map, characterize, and manipulate R genes of the cotton genome. This revised version was published online in July 2006 with corrections to the Cover Date.     

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.     

Cytogenetics of the 'glandless-seed and glanded-plant' trait from Gossypium sturtianum Willis introgressed into upland cotton (Gossypium hirsutum L.)

In order to introgress the ‘glandless-seed and glanded-plant’ trait from Gossypium sturtianum Willis (2n= 2x= 26, C₁ genome) into the cultivated upland cotton Gossypium hirsutum L. (2n= Ax= 52 (AD), genome), two trispecific hybrids have been created using either Gossypium thurberi Torado (2n= 2x= 26, D₁ genome) or Gossypium raimondii Ulbrich (2n= 2x= 26, D₅ genome) as bridge species. The cross of both trispecific hybrids by G. hirsutum produced the first backcross progenies (BCl). Cytogenetic analysis showed that the trispecific hybrids had 52 chromosomes, their chromosome configurations at metaphase I (Ml) being 15.071 + 15.3411 + 0.93III + 0.69IV + 0.26VI in G. thurberi×G. sturtianum×G. hirsutum (TSH) and 14.421 + 17.0311 + 0.82III + 0.15IV + 0.07VI in G. hirsutum × G. raimondii ×. G. sturtianum (HRS), respectively. Among six BCl plants analysed, the only plant expressing the ‘glandless-seed and glanded-plant’ trait had 52 chromosomes and a meiotic configuration of 5.261 + 20.61II + 0.69III + 0.77IV at MI. Pollen fertility was 2.90% in TSH, 8.97% in HRS, and ranged from 0% to 40.28% in the BCl progenies. The introgressed BCl plant is perennial in growth habit. It can be used in breeding programmes aiming at the introgression of the ‘glandless-seed and glanded-plant’ trait into a cultivar of upland cotton.     

Development of Gossypium hirsutum lines with cytoplasmic bollworm tolerance from Gossypium arboreum and Gossypium tomentosum

American cotton [Gossypium hirsutum (L.)] grown in India belongs to the race Latifolium. It is prone to bollworms [Helicoverpa armigera (Hubner), Erias vitella (Fabricius), Pectinophora gossypiella (Saunders)] infestation which causes considerable crop loss. Two breeding lines ‘Bikaneri (BN)‐arboreum (ARB)‐16’ and ‘BN‐tomentosum (TOM)‐277’ are cytoplasmic diverse genetic stocks. They were developed by crossing experimental lines ‘Delta Branch Experiment Station (DES)‐ARB16’ and ‘DES‐TOM277’ (non‐recurrent parents) with G. hirsutum‘BN’ (recurrent parent). Compared with BN these stocks possessed higher amount of gossypol, flavonol and phenol contents in leaves, stem and square. Plants were tolerant to bollworms and inherited comparable agronomic properties (yield, boll weight, plant height, number of monopods and sympods and fibre quality). Cotton breeders can use these lines for breeding cotton resistant to bollworm.     

QTL analysis for early-maturing traits in cotton using two upland cotton (Gossypium hirsutum L.) crosses

Making use of the markers linked closely to QTL for early-maturing traits for MAS (Marker-assisted selection) is an effective method for the simultaneous improvement of early maturity and other properties in cotton. In this study, two F₂ populations and their F_2:3 families were generated from the two upland cotton (Gossypium hirsutum L.) crosses, Baimian2 × TM-1 and Baimian2 × CIR12. QTL for early-maturing traits were analyzed using F_2:3 families. A total of 54 QTL (31 suggestive and 23 significant) were detected. Fourteen significant QTL had the LOD scores not only > 3 but also exceeding permutation threshold. At least four common QTL, qBP-17 for bud period (BP), qGP-17a/qGP-17b (qGP-17) for growth period (GP), qYPBF-17a/qYPBF-17b (qYPBF-17) for yield percentage before frost (YPBF) and qHFFBN-17 for height of first fruiting branch node (HFFBN), were found in both populations. These common QTL should be reliable and could be used for MAS to facilitate early maturity. The common QTL, qBP-17, had a LOD score not only > 3 but also exceeding permutation threshold, explaining 12.6% of the phenotypic variation. This QTL should be considered preferentially in MAS. Early-maturing traits of cotton are primarily controlled by dominant and over-dominant effects.     

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。     

陆地棉种质资源抗旱性状的关联分析

干旱是导致全世界棉花严重减产、纤维品质下降的重要因素,因此获得高产、优质、耐旱的棉花新品种一直是棉花的育种目标。本研究选取217份陆地棉栽培种组成的自然群体为研究对象,采用全生育期处理组灌水量为对照组50%的干旱胁迫处理,并在处理后期对217份材料的株高、衣分、单铃重等18个性状进行2年2点的表型鉴定,干旱胁迫后,群体间响应差异明显,多个表型性状在对照和处理间表现显著差异。通过BLUP分析表型数据并计算各性状的抗旱系数;全基因组范围选取的214对多态性SSR分子标记扫描群体,共检测到393个多态性位点,基因多样性系数平均值为0.402,范围为0.072～0.631, PIC值平均为0.329,范围为0.070～0.560;群体结构分析表明,该群体可分为2个亚群。用上述SSR标记分别对18个性状的抗旱系数进行关联分析,共关联到76个极显著位点(P<0.01),表型变异解释率为2.930%～7.218%,其中共有14个标记位点能同时被2种或以上性状检测到。研究结果可为后期棉花杂交育种亲本选择及抗旱分子标记辅助育种提供理论基础及参考依据。     

陆地棉产量相关性状的QTL定位

中棉所28和湘杂棉2号分别是以中棉所12×4133和中棉所12×8891配制而成的两个陆地棉强优势杂交种。以其F₂为作图群体,筛选6000多对SSR引物,利用两群体间27个共有多态位点,通过JoinMap 3.0软件整合了一张包含245个多态位点、全长1847.81 cM的遗传图谱。利用Win QTLCart 2.5复合区间作图法分别对两群体8个产量相关性状在F₂和F_2:3中进行QTL定位,在中棉所28群体多环境平均值的联合分析中检测到16个QTL,三环境分离分析中检测到43个QTL;在湘杂棉2号群体分别检测到20个和66个QTL。在A3、D8、D9等染色体上有QTL成簇分布现象,同时在两个群体中发现一些不受环境影响且稳定遗传的QTL。对考察的8个性状在两个群体中发现12对共有QTL,控制果枝数、衣分和籽指的QTL增效基因位点均来源于共同亲本中棉所12。综合分析推测中棉所12的育种价值主要是通过提高后代的结铃性来实现的。研究结果为棉花产量性状的分子设计育种提供了有用的信息。     

低酚棉棉子品质性状的遗传分析

利用ＡＤＭ模型对棉子品质性状的遗传分析结果表明，壳指、仁指、仁壳比、种仁率和种仁蛋白质指数存在母体效应，其中蛋白质指数的母体效应还大于加性和显性效应。子指主要受显性效应控制，且不存在母体效应，其它四个棉子物理品质性状的显性效应大于加性效应，但年度间存在差异。蛋白质含量、油分含量和油分指数不存在母体效应。所有棉子品质性状的狭义遗传率均较低。仁指可作为棉子物理品质性状改良的选择指标；棉子营养品质的选择在很大程度上取决于快速、可靠的检验方法的建立和完善。ＩＳＡＢＣ４ｇｌ可用于棉子物理品质性状的改良，浙棉９号可用于棉子营养品质性状的改良。这两个品种的杂种后代，棉子品质有较强的杂种优势。     

Phenotype Variations in Plant Regeneration of Upland Cotton (Gossypium hirsutum L.)

以陆地棉冀无2031无菌苗下胚轴为外植体，在MS中附加激素0.1mg/L IAA+0.1mg/L 2,4-D+0.1mg/L KT的培养基上诱导出愈伤组织。选择疏松愈伤组织继代到附加0.1mg/L ZT的培养基中，经2~3个月的继代培养诱导出颗粒状疏松胚性愈伤组织。胚性愈伤组织在无激素、每升加2g谷氨酰胺和0.5g天门冬酰胺的胚性愈伤增殖萌发培养基上培养2个月左右，可陆续形成不同时期的体胚苗。对冀无2031不同时期的体细胞萌发后的体胚苗进行观察，发现了真叶畸形苗、子叶节生根胚、胚轴侧生根胚、腋芽丛生苗和以腋     

Response to selective pressure in early generation progenies of upland cotton (Gossypium hirsutum L.)

Summary Combining high fiber strength with high yield in upland cotton (Gossipium hirsutum L.) was been difficult. The cross combination, 69–120 × 6M-10, was chosen for this study because of the divergence of parents for fiber strength, seeds per boll and crop maturity. Forty F₂ plants were selected in 1974 solely on the basis of visual yield and 20 plants were randomly chosen to serve as a bulk check. The F₃ progenies, the bulk check, the parents and a commercial check were tested in 1975. From these results, five groups were established, consisting of four (10%) progenies each to represent high seed/boll, low seeds/boll, high fiber strength, low fiber strength and high yield in the 1976 test of F₄ progenies. This experiment was conducted to (a) compare the contributions of yield components to lint yield among groups of early generation progenies (F₃ and F₄) and (b) determine the phenotypic correlations between yield and quality attributes. The yield components, bolls/m², seeds/boll, fibers/seed, mean fiber length and micronaire, were included as well as fiber strength.No significant lint yield differences were found among the five groups. Progenies within groups did, however, differ in lint yield. The low and high seeds boll groups gave similar yield and fiber quality results. The low and high fiber strength groups gave similar yield but longer fiber was obtained with the high strength group.Stepwise regression analyses estimates show that the number of bolls produced per unit area, although the major contributor to lint yield for all groups, contributed only 66.9% of the total variation in the progeny group selected for high yield, with fibers/seed, mean fiber length, micronaire and seeds/boll accounting for 13.2%, 8.1%, 6.0%, and 5.8%, respectively. Correlations between the various yield components and fiber strength were low and mainly insignificant, indicating that minor alterations might be made in later generations without serious consequences.Potential gains from second-stage selective pressure upon yield-per-boll components following initial selection for lint yield are discussed.     

陆地棉棕色纤维色泽的遗传效应

以2个棕色和3个白色纤维陆地棉做完全双列杂交,分析陆地棉棕色纤维的遗传效应、长绒与短绒的遗传相关及F1的色泽差异。用扫描仪获取长绒和短绒图像,利用Photoshop 9.0获取图像RGB信息、量化纤维色泽。按照QGAStation软件中的ADM和AD模型,采用MINQUE法分析,调整无偏预测法(AUP)预测各遗传效应值。结果表明,棕色棉的长绒和短绒的遗传规律一致,其加性和显性遗传方差均极显著,其中,长绒的加性遗传方差比率为0.8501,约为显性遗传方差比率的6倍,短绒的加性遗传方差比率为0.8726,约为显性遗传方差比率的8倍;相关分析显示长绒和短绒的基因型和表现型均达显著相关,基因型相关系数达0.9935;5个亲本加性效应均不相同,但均达极显著水平,其中,棕色棉为正效应,白色棉为负效应。说明棕色纤维陆地棉的长绒和短绒色泽的遗传变异主要来自加性和显性效应,其中加性效应起主导作用;长绒和短绒的色泽遗传存在连锁和互作;因不同品种(系)的加性效应大小不同,造成不同F1纤维色泽的表现差异。     

几种陆地棉腋芽快速繁殖的研究

以陆地棉的三个品种 (系 )棕色棉 ( X0 0 7)、绿色棉 ( X0 0 8)和白色棉 coker31 2为材料 ,进行不同培养条件下棉花幼苗腋芽快速繁殖的比较试验。分析了不同棉花品种 (系 )基因型、不同培养基类型、以及不同无菌苗苗龄对腋芽快速繁殖的影响因子 ,初步建立了陆地棉腋芽的诱导、伸长、生根、壮苗和移栽的技术体系 ,并将该技术体系在转acs B基因彩色棉品系的扩繁上得到了较理想的应用     

Leaf Growth, Reproduction Growth and Yield in Cotton (Gossypium hirsutum L.)

In three genotypes of cotton differing in height, reproductive growth increased with the increase in number and size of the leaves and was maximum when leaf area was also maximum. After this stage though the leaf area continued to remain high for 3 weeks and then declined, more buds were shed than produced. Within genotypes the taller type LRA 5166 produced more leaves with greater area than MCU-5 whereas the dwarf type KD (CAKD) had more leaves with lesser area than MCU-5. The maximum leaf area as well as the yield of seed cotton were one and a half times in LRA 5166 than in MCU-5. Though the maximum leaf area in MCU-5 was also one and a half times than in KD (CAKD) , their seed cotton yields were the same. Since even during the period of maximum reproductive growth substantially more number of leaves were produced by LRA 5166 and KD (CAKD) than MCU-5 , maintenance and retention of bolls appeared to depend not only on leaf number and area but on their photosynthetic efficiency and perhaps nutrient reserves.     

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...     

陆地棉种子物理性状QTL定位

定位棉花种子性状的基因对揭示棉花种子性状的遗传规律,以及明确棉花种子、产量、纤维品质等性状间的遗传关系具有重要意义。以(渝棉1号×T586) F_2:7重组近交系群体构建的遗传连锁图谱,在鉴定270个家系3个环境种子物理性状的基础上,利用MQM作图方法,共检测到34个种子物理性状QTL,包括9个种子重(qSW)、5个短绒重(qFW)、3个短绒率(qFP)、8个种仁重(qKW)、6个种子壳重(qHW)和3个种仁率(qKP)QTL,它们可解释4.6%~80.1%的性状表型变异。9个QTL在2个或3个环境中被检测到,其中包括第12染色体显性光子位点的短绒重与短绒率QTL,以及另外7个微效应QTL。34个QTL分布于15条染色体,其中A染色体组20个,D染色体组14个。有12个染色体区段分布有2个或2个以上的QTL,而且同一染色体区域同一亲本所具有的不同性状QTL的方向大多数与性状表型相关系数的正负一致。     

陆地棉体细胞再生植株技术的改进研究

以Coker201为材料,改进了棉花体细胞再生植株及移栽技术。MSB(MS无机盐+B5维生素)培养基附加IBA能直接诱导出胚性愈伤组织,适当浓度的KT可促进IBA的效应。最适激素组合(IBA1.0mg·L-1、KT0.5mg·L-1)能使几个陆地棉品种的下胚轴在两个月内诱导出大量胚性愈伤组织,诱导后第三个月可观察到胚状体的发生。适量的AgNO3能促进愈伤组织的增殖。继代时将激素降为IBA0.5mg·L-1、KT0.1～0.2mg·L-1,有利于胚状体发育长大。增加KNO31.9g·L-1,同时使用蔗糖和葡萄糖各15g·L-1作碳源有利于胚状体的分化。采用含0.4g·L-1活性碳的MS为成苗培养基,并结合1/6MS液培诱导生根技术,使苗粗壮易于移栽。从愈伤组织诱导至植株再生,约需5～6个月时间。     

Association mapping for seed cotton yield,yield components and fibre quality traits in upland cotton (Gossypium hirsutum L.) genotypes

Association mapping has revolutionized human genetics and is increasingly used in plant genetics. It is an efficient way of determining the genetic basis of complex traits. In cotton so far numerous association mapping studies are available as compared to many other important crops. In our study, mapping was performed using cotton 63K infinium beadchip with 201 germplasm lines. Through fast STRUCTURE analysis, lines were grouped into 12 subgroups and revealed genome sharing among the groups. The critical value (R²) was set to 0.243 as threshold to claim LD between two loci. About 3.13% marker pairs recorded significantly high LD (R² = 1), and 8.19% of marker pairs were in the range of 0.3 to 0.99 R². In MLM, 349 significant marker–trait associations were detected as against 642 in GLM because of effectiveness in eliminating false associations in MLM. A total of 68 markers explained >10% phenotypic variation for yield and fibre quality traits. Phenotypic variation explained by markers was smaller, suggesting that they might be associated with minor QTLs.