油菜油分、蛋白质和硫苷含量相关性分析及QTL 定位

为定位与油分、蛋白质和硫苷含量等品质性状相关的数量性状位点(QTL), 以2个含油量较高的甘蓝型油菜(Brassica napus)品系8908B和R1为研究材料, 配置正反交组合。在正反交F₂代群体中, 含油量和蛋白质含量都存在极显著的负相关, 相关系数分别为-0.68和-0.81, 含油量和硫苷含量相关性不显著; 蛋白质含量和硫苷含量在正交群体中相关性不显著, 但在反交群体中存在显著负相关(相关系数r =-0.45)。利用正交F₂代群体中的118个单株, 构建了包含121个标记的遗传连锁图谱, 图谱长1 298.7 cM, 有21个连锁群(LGs)。采用复合区间作图法, 在连锁图上定位了2个与含油量有关的QTL, 分别位于LG8和LG10, 其贡献率分别为4.8%和13.7%, 增效基因都来源于R1; 定位了2个与蛋白质含量有关的QTL: pro1 和 pro2, 分别位于LG1和LG3, 其贡献率分别为15.2%和14.1%, 位点pro1由8908B提供增效基因, pro2则由R1提供增效基因; 定位了4个与硫苷含量有关的QTL, 其中LG20上有2个, LG4和LG8上各1个, 它们的贡献率在1.9%-25.4%之间, 除LG20上glu1的增效基因来自R1外, 其余3个QTL位点均由8908B提供增效基因。     

高丹草氢氰酸含量性状的QTL定位分析

为了深入开展高丹草低氢氰酸含量性状的QTL精细定位、基因图位克隆、功能解析及分子标记辅助育种,该研究以二倍体杂交高丹草F_2代群体500个分离单株无性系及其亲本为材料,在课题组前期已构建出的高丹草高密度分子遗传连锁图谱的基础上,利用区间作图法对两年两地测定的高丹草氢氰酸含量性状进行了QTL定位分析。结果显示:(1)在4个不同环境下高丹草氢氰酸含量性状的广义遗传率分别为61.70%、72.05%、40.16%和69.25%,表明氢氰酸含量是既受环境影响又受微效多基因控制的数量性状,而且其群体测定值频率呈明显单峰正态性分布特点,符合QTL定位要求。(2)在LOD2.5的条件下,共检测到16个与氢氰酸含量性状相关的QTLs,其分布在LG1、LG2、LG4、LG6、LG7、LG8和LG10连锁群上。(3)16个QTLs中能重复检测到的稳定QTLs有9个,遗传贡献率范围为1.17%～39.9%,其中贡献率大于20%的主效QTLs有Qcn2-2、Qcn4-1、Qcn6和Qcn6-1共4个。该研究结果明确了各QTLs的遗传效应和分子标记位点。     

甜瓜果实糖含量相关性状QTL分析

以高糖栽培亲本自交系‘0246’为母本,低糖野生亲本自交系‘Y101’为父本,通过杂交得到了含135个单株的甜瓜远缘F2群体,分别测定甜瓜果实果糖、葡萄糖和蔗糖含量,将三者之和作为总糖含量,进行遗传连锁图谱构建及QTL分析。结果表明:(1)构建的甜瓜果实遗传图谱包含14个连锁群,覆盖基因组长度726.30cM,标记间平均距离为12.74cM。(2)检测到与总糖含量和果糖含量相关的QTL位点各1个,分别命名为Ts3.1和Fru4.1,贡献率分别为14.89%和13.02%,分布于第3、4连锁群,LOD值分别为3.60和3.10。2个QTL位点均为正向加性遗传,分别对增加总糖和果糖含量表现为增效累加效应。研究结果为开展甜瓜糖含量相关基因精细定位和克隆研究奠定了基础。     

水稻籽粒锌含量的QTL定位

锌元素的营养失衡已成为影响人类健康的最重要因素之一,籽粒锌含量的QTL（quantitative trait loci）定位对研究富锌水稻的遗传育种具有重要的意义。以水稻（Oryzasativa L．）亲本奉新红米和明恢100杂交的145个株系的F2群体为实验材料,利用92个SSR（simple sequence repeat）标记对水稻籽粒锌含量进行了QTL定位,共检测到3个QTLs,分别定位于第3、6和11染色体上,对表型变异的贡献率分别为4．97％、12．75％和7．74％。其中位于第3染色体上的分子标记RM186和RM168之间的QZN3对表型变异的贡献率最大,其增效等位基因来自亲本明恢100,表现为部分显性。3个QTLs的联合贡献率为25．46％。具有基因累加效应。该研究结果有利于深入理解水稻锌含量的遗传基础,为锌含量的QTL精细定位、基因克隆和分子标记辅助选择提供依据。     

大豆遗传图谱的构建和若干农艺性状的QTL定位分析

大豆许多重要农艺性状都是由微效多基因控制的数量性状,对这些数量性状进行QTL定位是大豆数量性状遗传研究领域的一个重要内容.本研究利用栽培大豆科新3号为父本、中黄20为母本杂交得到含192个单株的F2分离群体,构建了含122 个SSR标记、覆盖1719.6cM、由33个连锁群组成的连锁遗传图谱.利用复合区间作图法,对该群体的株高、主茎节数、单株粒重和蛋白质含量等农艺性状的调查数据进行QTL分析,共找到两个株高QTL,贡献率分别为9.15%和6.08%;两个主茎节数QTL,贡献率分别为10. 1%和8.6%;一个蛋白质含量QTL,贡献率为9.8%;一个单株粒重QTL,贡献率为11.4% .通过遗传作图共找到与所定位的4个农艺性状QTL连锁的6个SSR标记,这些标记可以应用于大豆种质资源的分子标记辅助选择,从而为大豆分子标记辅助育种提供理论依据.     

中国对虾抗WSSV及其它经济性状的QTL定位初步研究

以中国对虾抗WSSV选育群体第四代雌虾和野生中国对虾雄虾为亲本,采用人工精荚移植方式产生F1代家系,家系内个体姊妹交获得R家系材料,42尾R家系个体采用口饲法进行WSSV（White Spot Syndrome Virus）攻毒实验,获得个体抗WSSV及其它相关数据。构建了中国对虾的AFLP（Amphfied Fragment Length Polymorphism）分子标记遗传连锁图谱。利用MAPMAKER／QTL1．1软件进行了中国对虾体长、全长、体重及抗WSSV性状的QTL（Quantitative TraitsLoci）定位分析,首次实现了中国对虾重要经济性状的QTL定位。在LOD值大于2．0的条件下,共检测到和体长相关的QTL位点1个,与全长相关的QTL位点2个,与体重相关的QTL位点2个,与抗WSSV性状相关的位点2个,分别位于3个连锁群上,位点变异解释率从26．6％-66．9％不等。在其中的1个连锁群上检测到了体重、全长和抗WSSV性状相关的三个QTL位点,1个连锁群上检测到了体重和抗WSSV性状相关的两个QTL位点,1个连锁群上检测到了全长和体长相关的两个QTL位点。表明在中国对虾在此生长阶段,抗WSSV性状和个体大小存在一定程度的正相关关系[动物学报54（6）：1075-1081,2008]。     

黄瓜SRAP遗传连锁图的构建及侧枝基因定位

在由黄瓜的两个自交系S06与S52杂交产生的F₂群体中, 应用SRAP(sequence-related amplified polymorphism)标记构建遗传连锁图谱, 检测控制黄瓜侧枝数(lbn)和侧枝平均长度(lbl)的数量性状座位(QTL). 使用筛选出的64个多态性引物组合对F2群体进行分析, 得到108个多态性位点. 经MAPMAKER/EXP3.0分析(LOD≥3.0), 获得由92个标记座位组成、覆盖7个连锁群的遗传图谱, 总长1164.2 cM, 标记平均间距12.6 cM. 应用QTLMapper1.6, 各检测到4个控制lbn和lbl的QTL, 其中, 对lbn贡献率最大的QTL位于第二连锁群的ME11SA4B-ME5EM5区间, 其S06基因型具有增效作用; 对lbl贡献率最大的QTL位于第二连锁群的DC1OD3-DC1EM14区间, 其S06基因型具有增效作用.     

谷子SSR分子图谱的构建及主要农艺性状QTL定位

试验拟对谷子重要农艺性状进行数量性状位点QTL分析。以表型差异较大的沈3/晋谷20F2作图群体为材料,观测其株高、穗长等性状,选用SSR做分子标记,利用完备区间作图法(BASTEN C J)进行QTL分析。结果显示,表型数据在作图群体中呈现连续分布,表现为多基因控制的数量性状,被整合的54个SSR标记构建10个连锁群,LOD阈值设置为2.0,检测到与株高相关的主效QTL2个,联合贡献率45.9637%,穗长主效QTL1个,贡献率14.9647%,与穗重、粒重相关的主效QTL为同一位点,贡献率分别为11.9601%和10.1879%。有6组QTL位点之间存在基因互作效应,大小范围为-0.4986-16.6407,对性状的贡献率在2.2716％至6.7478％之间。谷子表型控制复杂,相关QTL的检测受环境影响较大,不同连锁群QTL间互作明显。     

甘蓝分子连锁图的构建与品质性状的QTL定位

以两个不同生态型甘蓝(Brassica oleracea var．capitata)品种杂交得到的F2代为作图群体,用RAPD标记构建甘蓝分子连锁图。通过对520个随机引物进行筛选,236个引物在两亲本间表现多态性,多态性比例为47．7％。选取111个引物对群体进行分析,构建了一张含有135个标记位点,9个连锁群,覆盖长度为1023．7cM的分子连锁图。利用该图谱对甘蓝叶球紧实度和中心柱长两性状进行了QTL定位分析。检测到3个与叶球紧实度相关的QTL,总贡献率为62．5％;检测到4个与中心柱长相关的QTL,总贡献率为59．1％。     

亚麻主要农艺与品质相关性状的QTL定位

为了全面了解亚麻产量和品质相关性状的遗传基础,为亚麻基因克隆和分子标记辅助育种提供理论依据,在已构建SNP连锁遗传图谱的基础上,以LH-89为父本,R43为母本构建F2:3家系QTL定位群体,用R/QTL软件采用复合区间作图法对13个农艺和品质性状进行QTL定位。结果表明:(1)该研究共检测出35个QTL位点,与粗脂肪及其组成成分相关的QTL有20个,与农艺性状相关的QTL有15个;其中:亚油酸和粗脂肪各5个,亚麻酸、千粒重各4个,棕榈酸、株高、工艺长度各3个,硬脂酸、分枝数各2个,单株果数、果粒数、单株粒重、油酸各1个。(2)共有18个QTL的表型贡献率超10%(主效基因),其中农艺性状定位8个主效基因,品质性状定位10个主效基因。     

Homoeologous loci control the accumulation of seed glucosinolates in oilseed rape (Brassica napus).

The genetic control of seed glucosinolate content in oilseed rape was investigated using two intervarietal backcross populations. Four QTLs segregating in the population derived from a Brassica napus L. 'Victor' x Brassica napus L. 'Tapidor' cross, together accounting for 76% of the phenotypic variation, were mapped. Three of these loci also appeared to control the accumulation of seed glucosinolates in a Brassica napus L. 'Bienvenu' x 'Tapidor' cross, and accounted for 86% of the phenotypic variation. The three QTLs common to both populations mapped to homoeologous regions of the B. napus genome, suggesting that seed glucosinolate accumulation is controlled by duplicate genes. It was possible to extend the comparative analysis of QTLs controlling seed glucosinolate accumulation by aligning the published genetic maps generated by several research groups. This comparative mapping demonstrated that high-glucosinolate varieties often carry low-glucosinolate alleles at one or more of the loci controlling seed glucosinolate accumulation.     

Genetic linkage map construction and QTL mapping of cadmium accumulation in radish (Raphanus sativus L.)

Cadmium (Cd) is a widespread soil pollutant and poses a significant threat to human health via the food chain. Large phenotypic variations in Cd concentration of radish roots and shoots have been observed. However, the genetic and molecular mechanisms of Cd accumulation in radish remain to be elucidated. In this study, a genetic linkage map was constructed using an F(2) mapping population derived from a cross between a high Cd-accumulating cultivar NAU-Dysx and a low Cd-accumulating cultivar NAU-Yh. The linkage map consisted of 523 SRAP, RAPD, SSR, ISSR, RAMP, and RGA markers and had a total length of 1,678.2 cM with a mean distance of 3.4 cM between two markers. All mapped markers distributed on nine linkage groups (LGs) having sizes between 134.7 and 236.8 cM. Four quantitative trait loci (QTLs) for root Cd accumulation were mapped on LGs 1, 4, 6, and 9, which accounted for 9.86 to 48.64 % of all phenotypic variance. Two QTLs associated with shoot Cd accumulation were detected on LG1 and 3, which accounted for 17.08 and 29.53 % of phenotypic variance, respectively. A major-effect QTL, qRCd9 (QTL for root Cd accumulation on LG9), was identified on LG 9 flanked by NAUrp011_754 and EM5me6_286 markers with a high LOD value of 23.6, which accounted for 48.64 % of the total phenotypic variance in Cd accumulation of F(2) lines. The results indicated that qRCd9 is a novel QTL responsible for controlling root Cd accumulation in radish, and the identification of specific molecular markers tightly linked to the major QTL could be further applied for marker-assisted selection (MAS) in low-Cd content radish breeding program.     

Construction of a High-Density Microsatellite Genetic Linkage Map and Mapping of Sexual and Growth-Related Traits in Half-Smooth Tongue Sole (Cynoglossus semilaevis)

High-density genetic linkage maps of half-smooth tongue sole were developed with 1007 microsatellite markers, two SCAR markers and an F1 family containing 94. The female map was composed of 828 markers in 21 linkage groups, covering a total of 1447.3 cM, with an average interval 1.83 cM between markers. The male map consisted of 794 markers in 21 linkage groups, spanning 1497.5 cM, with an average interval of 1.96 cM. The female and male maps had 812 and 785 unique positions, respectively. The genome length of half-smooth tongue sole was estimated to be 1527.7 cM for the females and 1582.1 cM for the males. Based on estimations of the map lengths, the female and male maps covered 94.74 and 94.65% of the genome, respectively. The consensus map was composed of 1007 microsatellite markers and two SCAR markers in 21 linkage groups, covering a total of 1624 cM with an average interval of 1.67 cM. Furthermore, 159 sex-linked SSR markers were identified. Five sex-linked microsatellite markers were confirmed in their association with sex in a large number of individuals selected from different families. These sex-linked markers were mapped on the female map LG1f with zero recombination. Two QTLs that were identified for body weight, designated as We-1 and We-2, accounted for 26.39% and 10.60% of the phenotypic variation. Two QTLs for body width, designated Wi-1 and Wi-2, were mapped in LG4f and accounted for 14.33% and 12.83% of the phenotypic variation, respectively. Seven sex-related loci were mapped in LG1f, LG14f and LG1m by CIM, accounting for 12.5–25.2% of the trait variation. The results should prove to be very useful for improving growth traits using molecular MAS.     

四倍体栽培棉种产量和纤维品质性状的QTL定位

陆地棉和海岛棉是两个不同的四倍体栽培种 ,但在生产上各有其特点 ,陆地棉丰产性强 ,海岛棉纤维品质优良 ,利用其种间杂交群体定位产量和品质性状的QTL ,对于分子标记辅助的海岛棉优质纤维向陆地棉转移很有意义。以SSR和RAPD为分子标记 ,陆地棉与海岛棉杂种 (邯郸 2 0 8×Pima90 )F2 群体为作图群体 ,构建了一张含 12 6个标记的遗传图谱 ,包括 6 8个SSR标记和 5 8个RAPD标记 ,可分为 2 9个连锁群 ,标记间平均距离为 13 7cM ,总长1717 0cM ,覆盖棉花总基因组约 34 34% ;以遗传图 12 6个标记为基础 ,对F2 :3 家系符合正态分布的 10个农艺性状及纤维品质性状进行全基因组QTL扫描 ,结果发现 2 9个QTL分别与产量和品质性状有关。其中与衣指、籽指、皮棉产量、子棉产量、衣分等产量性状相关的QTL分别有 1、3、5、6和 1个 ,与纤维长度、整齐度、强度、伸长率和马克隆值等品质性状相关的QTL分别有 2、4、2、4和 1个。各QTL解释的变异量在 12 4 2 %～ 47 0 1%之间。其中比强度有关的 2个QTL能够解释的表型变异率分别为 34 15 %和 13 86 %。     

Genetic mapping of QTLs conditioning soybean sprout yield and quality

Soybean sprouts have been used as a food in the Orient since ancient times. In this study, 92 restriction fragment length polymorphism (RFLP) loci and two morphological markers (W1 and T) were used to identify quantitative trait loci (QTLs) associated with soybean sprout-related traits in 100 F₂-derived lines from the cross of ’Pureunkong’×’Jinpumkong 2’. The genetic map consisted of 76 loci which covered about 756 cM and converged into 20 linkage groups. Eighteen markers remained unlinked. Phenotypic data were collected in 1996 and 1997 for hypocotyl length, percentage of abnormal seedlings, and sprout yield 6 days after germination at 20°C. Hypocotyl length was determined as the average length from the point of initiation of the first secondary root to the point of attachment of the cotyledons. The number of decayed seeds and seedlings, plus the number of stunted seedlings (less than 2-cm growth), was recorded a s abnormal seedlings. Seed weight was determined based on the 50-seed sample. Sprout yield was recorded as the total fresh weight of soybean sprouts produced from the 50-seed sample divided by the dry weight of the 50-seed sample. Four QTLs were associated with sprout yield in the combined analysis across 2 years. For the QTL linked to L154 on the Linkage Group (LG) G the positive allele was derived from Pureunkong (R ² = 0.19), whereas at the other three QTLs (A089 on LG B1, A668n on LG K and B046 on LG L) the positive alleles were from Jinpumkong 2. QTLs conditioning seed weight were linked to markers A802n (LG B1), A069 (LG E), Cr321 (LG F) and A235 (LG G). At these four markers, the Jinpumkong allele increased seed weight. Markers K011n on LG B1, W1 on LG F and A757 on LG L were linked to QTLs conditioning hypocotyl length; and Bng119, K455n and K418n to QTLs conditioning the abnormal seedlings. The QTLs conditioning sprout yield were in the same genomic locations as the QTLs for seed weight identified in this population or from previously published research, indicating that QTLs for sprout yield are genetically linked to seed-weight QTLs or else that seed-weight QTLs pleiotropically condition sprout yield. These data demonstrate that effective marker-assisted selection may be feasible for enhancing sprout yield in a soybean. The transgressive segregation of sprout yield, as well as the existence of two QTLs conditioning greater than 10% of the phenotypic variation in sprout yields provides an opportunity to select for progeny lines with a greater sprout yield than currently preferred cultivars such as Pureunkong. Received: 23 August 2000 / Accepted: 23 January 2001     

Mapping of Quantitative Trait Loci for Butter Content and Hardness in Cocoa Beans (Theobroma cacao L.)

Cocoa butter is an important raw material for the chocolate, pharmaceutical, and cosmetic industries. The butter content and quality in cocoa beans are genetically controlled characteristics, and affect its commercial value and industrial applicability. In the present work, an F₂ population derived from the cross between the ICS-1 and Scavina-6 cocoa clones was used for molecular mapping. A linkage map was constructed based on amplified fragment length polymorphism, random amplified polymorphic DNA, and simple sequence repeat markers, resulting in a total of 273 markers, distributed in 14 linkage groups (LGs). Phenotyping of butter content was performed after ether extraction and butter hardness was determined by sweeping differential calorimetry. One quantitative trait locus (QTL) associated to butter content was mapped at linkage group 9 (LG9) and two QTLs for butter hardness were identified at linkage groups 9 and 7 (LG9 and LG7). The two QTLs mapped at the LG9 explained 51.0% and 28.8% of the phenotypic variation for butter content and hardness, respectively. These QTLs were concentrated in the same map region, suggesting a close genetic linkage or pleiotropic effect. The QTLs identified may be useful in further marker-assisted selection breeding programs aimed at cocoa butter quality improvement.     

Identification of quantitative trait loci for plant height, lodging, and maturity in a soybean population segregating for growth habit

The use of molecular markers to identify quantitative trait loci (QTLs) has the potential to enhance the efficiency of trait selection in plant breeding. The purpose of the present study was to identify additional QTLs for plant height, lodging, and maturity in a soybean, Glycine max (L.) Merr., population segregating for growth habit. In this study, 153 restriction fragment length polymorphisms (RFLP) and one morphological marker (Dt1) were used to identify QTLs associated with plant height, lodging, and maturity in 111 F₂-derived lines from a cross of PI 97100 and Coker 237. The F₂-derived lines and two parents were grown at Athens, Ga., and Blackville, S.C., in 1994 and evaluated for phenotypic traits. The genetic linkage map of these 143 loci covered about 1600 cM and converged into 23 linkage groups. Eleven markers remained unlinked. Using interval-mapping analysis for linked markers and single-factor analysis of variance (ANOVA), loci were tested for association with phenotypic data taken at each location as well as mean values over the two locations. In the combined analysis over locations, the major locus associated with plant height was identified as Dt1 on linkage group (LG) L. The Dt1 locus was also associated with lodging. This locus explained 67.7% of the total variation for plant height, and 56.4% for lodging. In addition, two QTLs for plant height (K007 on LG H and A516b on LG N) and one QTL for lodging (cr517 on LG J) were identified. For maturity, two independent QTLs were identified in intervals between R051 and N100, and between B032 and CpTI, on LG K. These QTLs explained 31.2% and 26.2% of the total variation for maturity, respectively. The same QTLs were identified for all traits at each location. This consistency of QTLs may be related to a few QTLs with large effects conditioning plant height, lodging, and maturity in this population.     

Mapping QTLs for root morphological traits inBrassica rapa L. based on AFLP and RAPD markers

Root growth and thickening plays a key role in the final productivity and even the quality of storage roots in root crops. This study was conducted to identify and map quantitative trait loci (QTLs) affecting root morphological traits in Brassica rapa by using molecular markers. An F2 population was developed from a cross between Chinese cabbage (Brassica rapa ssp. chinensis) and turnip (B. rapa ssp. rapifera), which differed greatly in root characters. A genetic map covering 1837.1 cM, with 192 marker loci and 11 linkage groups, was constructed by using this F2 population. The F3 families derived from F2 plants were grown in the field and evaluated for taproot traits (thickness, length, and weight). QTL analysis via simple interval mapping detected 18 QTLs for the 3 root traits, including 7 QTLs for taproot thickness, 5 QTLs for taproot length, and 6 QTLs for taproot weight. Individually, the QTLs accounted for 8.4-27.4% of the phenotypic variation. The 2 major QTLs, qTRT4b for taproot thickness and qTRW4 for taproot weight, explained 27.4% and 24.8% of the total phenotypic variance, respectively. The QTLs for root traits, firstly detected in Brassica crops, may provide a basis for marker-assisted selection to improve productivity in root-crop breeding.     

Seed and agronomic QTL in low linolenic acid, lipoxygenase-free soybean (Glycine max (L.) Merrill) germplasm.

Linolenic acid and seed lipoxygenases are associated with off flavours in soybean products. F5 recombinant inbred lines (RILs) from a cross between a low linolenic acid line (RG10) and a seed lipoxygenase-free line (OX948) were genotyped for simple sequence repeats (SSR), random amplified polymorphic DNA (RAPD), sequence-tagged sites (STS), and cleaved amplified polymorphic sequence (CAPS) markers and evaluated for seed and agronomic traits at 3 Ontario locations in 2 years. One hundred twenty markers covering 1247.5 cM were mapped to 18 linkage groups (LGs) in the soybean composite genetic map. Seed lipoxygenases L-1 and L-2 mapped as single major genes to the same location on LG G13-F. L-3 mapped to LG G11-E. This is the first report of a map position for L-3. A major quantitative trait locus (QTL) associated with reduced linolenic acid content was identified on LG G3-B2. QTLs for 12 additional seed and agronomic traits were detected. Linolenic acid content, linoleic acid content, yield, seed mass, protein content, and plant height QTL were present in at least 4 of 6 environments. Three to 8 QTLs per trait were detected that accounted for up to 78% of total variation. Linolenic acid and lipoxygenase loci did not overlap yield QTL, suggesting that it should be possible to develop high-yielding lines resistant to oxidative degradation by marker-assisted selection (MAS).