胡萝卜(Daucus carota L.)中主要胡萝卜素和番茄红素含量的QTL分析

以高胡萝卜素自交系P50006和HCM A.C.为亲本构建的F2群体为作图群体,对胡萝卜中α-胡萝卜素、α-胡萝卜素、总胡萝卜素和番茄红素含量进行QTL定位及遗传分析。结果表明,α、β-胡萝卜素、总胡萝卜素和番茄红素含量的广义遗传力分别为0.75、0.50、0.31和0.93。遗传图谱包含91个SRAP(Sequence-related amplified polymorphism)标记,分布于9个连锁群,总长度502.9cM,标记间平均距离5.5cM。除α-胡萝卜素含量外,α-胡萝卜素、总胡萝卜素和番茄红素含量分别检测到1个主效QTL,均为加性遗传效应,分别解释表型变异为12.79%、12.87%和14.61%。此外,α-胡萝卜素和番茄红素含量还分别检测到1对上位性QTL,最大遗传效应分别为显性×加性互作和显性×显性互作,分别解释表型变异为15.1%和6.5%。文章中与QTL连锁的分子标记可用于高胡萝卜素、番茄红素的种质筛选和聚合育种。     

甘蓝型油菜千粒重性状的QTL初步定位研究

千粒重是油菜重要的产量相关性状之一,构建油菜遗传连锁图谱是研究其产量性状基因的前提。本研究利用小孢子培养技术,选育出了甘蓝型油菜大粒品系(G-42)和小粒品系(7-9)的纯合DH系DH-G-42和DH-7-9,其千粒重分别为6.24 g和2.42 g,二者比值达2.58。以DH-G-42为母本、DH-7-9为父本,构建了含190个单株的F2遗传作图群体,利用SSR和SRAP标记技术绘制遗传连锁图谱,该图谱共包含20个连锁群,涉及128个SSR标记和100个SRAP标记,图谱总长1546.6cM,标记间平均图距为6.78cM。本研究共检测到3个与千粒重性状相关的QTL,分别位于A9和C1连锁群,其中qSW-A9-1和qSW-A9-2贡献率分别达到10.98%和27.45%,均可视为控制粒重的主效QTL。本研究为后续进行油菜千粒重性状QTL的精细定位分析、分子标记辅助选择育种及新基因的克隆等奠定了基础。     

甜瓜遗传图谱的构建及果实与种子QTL分析

以遗传性状差异较大的甜瓜材料日本安农二号与新疆哈密瓜K413杂交产生的143个F2单株为作图群体,以AFLP与SSR分子标记为主构建了包含12个连锁群、142个遗传标记位点的甜瓜遗传图谱,其中包括121个AFLP标记、16个SSR标记、3个STS标记、2个性状标记,连锁群总长度为1 014.2 cM。应用复合区间作图法对甜瓜果实的大小、长宽比、糖度、硬度以及甜瓜种子的长、宽、形状、重量等性状进行遗传定位与分析。基因定位结果显示控制果肉颜色的基因位于C9连锁群AFLP分子标记NDAA与NCFA之间。其他性状表现为数量性状控制,共检测到25个数量性状基因座,不同性状基因座位有重叠分布的特点。其中C5连锁群标记NCA-N73C区间检测到QTLs Sl5.1、Sw5.1和Swt5.1分别控制种子长、宽和千粒重,分别可解释表型变异的17%、19%和23%。该区域包含的来自母本安农二号的基因位点对甜瓜种子的长、宽、千粒重均有明显的抑制作用;位于C8连锁群标记N73A与NFDA间的QTL通过影响种子的宽度从而影响种子的形状与重量;同样位于C8连锁群的果实长宽比QTL Fs8.1在F2和F3中均检测到,分别解释表型变异的25%和19%,表现为部分显性,来自安农二号的等位基因抑制甜瓜果实伸长,生成圆形甜瓜;还发现控制甜瓜果实心糖、边糖、果实硬度的QTL各一个。     

甘蓝型黄籽油菜种皮色泽QTL作图

甘蓝型黄籽油菜具有低纤维、高蛋白及高含油量的优点,因而己成为广大油菜育种工作者研究的重点之一。利用甘蓝型黑籽品系油研2号作父本,计蓝型黄籽品系GH06为母本,获得132个单株的F2群体;以AFLP和SSR为主要分析方法,构建了包括164个标记的甘蓝型油菜遗传连锁图谱,其中包括125个AFLP标记、37个SSR标记及一个RAPD和一个SCAR标记,分布在19个连锁群上,覆盖油菜基因组2549．8cM,标记间平均距离15．55cM。利用多区间作图法,对种皮色泽QTL进行分析,在第5及第19连锁群上各检测到一个QTL位点,分别解释表型变异46％及30．9％。     

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 %。     

基于SRAP分子标记的冰草遗传连锁图谱构建

构建高密度遗传连锁图谱是冰草抗性、品质、产量等重要性状QTL精细定位及标记辅助育种研究的基础。该试验以四倍体杂交冰草F2群体的202个分离单株及其亲本为材料,利用SRAP分子标记技术和Join Map 4.0作图软件对冰草的遗传连锁图谱进行了构建。结果表明:(1)共筛选出22对多态性好、标记位点清晰稳定的SRAP适宜引物,对冰草杂种F2分离单株的基因组DNA进行PCR扩增,共获得510个SRAP多态性标记位点,其比率占88.2%。(2)偏分离分析表明,偏分离标记比率仅为14.12%,符合遗传作图的要求。(3)成功构建了冰草的SRAP分子标记遗传连锁图谱,该图谱有14个连锁群、510个标记,连锁群间长度范围86.4～179.0cM,覆盖基因组总长度1 912.9cM,标记间平均间距3.75cM,为高密度遗传图谱。     

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。     

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.     

陆地棉遗传图谱构建与纤维品质性状QTL定位

以陆地棉(Gossypium hirsutum L.)品种Bar19/1和Acala1517-77杂交的108个F2单株为材料,应用85个标记(70个SSR标记和15个AFLP标记)构建了总长为814cM的遗传图谱,覆盖棉花基因组的18.3%。该图谱包含25个连锁群,分别对应到17条染色体和4个未知连锁群。应用复合区间作图法分析了该组合的108个F2单株和F3家系纤维品质性状,从遗传图谱上检测到19个纤维品质数量性状基因座(QTL),包括5个纤维长度、6个纤维比强度、4个伸长率及4个马克隆值QTL,分别解释各性状表型变异的15.11%~28.45%、8.46%~24.51%、11.08%~27.55%和9.23%~42.21%。纤维长度和伸长率的QTL以部分显性为主,少数具有超显性,比强度QTL以加性和部分显性为主,4个马克隆值QTL中有3个表现为超显性。研究结果表明,陆地棉Bar19/1和Acala1517-77间多态性位点丰富,有利于构建高密度遗传图谱,纤维品质性状的QTL分析从分子水平上揭示了纤维品质的遗传基础。     

甘蓝型油菜千粒重性状的QTL定位分析

该研究利用油菜双单倍体株系(348份)群体和已构建的遗传连锁图谱,采用复合区间作图法,对2009~2013年连续5年的千粒重性状表型数据进行QTL初步定位和分析,结果共获得46个显著性千粒重QTL,主要分布在A7、C1和C6等11条染色体上;其中qTSW-09 DL11-1的表型变异最高(19.63%),qTSW-11 DL9的表型变异最小(2.73%)。通过元分析方法将所获得的46个QTL进行整合,结果显示:cqTSW-C1-2的表型变异最大(10.64%),并发现多个整合后的一致性QTL能够在连续多年试验中被检测到,其中cqTSW-C1-3连续5年被检测到,表明控制千粒重的QTL在种植环境中能够稳定表达;同时,新发现位于C1染色体上的千粒重主效QTL cqTSW-C1-2,解释表型变异达到10.64%。油菜千粒重性状的QTL分析和主效QTL的获得,为进一步实现油菜大籽粒的分子育种和高产新品种的培育提供了重要的理论指导。     

Identification of a major QTL for silique length and seed weight in oilseed rape (Brassica napus L.)

Silique length (SL) and seed weight (SW) are two important yield-related traits controlled by quantitative trait loci (QTL) in oilseed rape (Brassica napus L.). The genetic bases underlying these two traits are largely unknown at present. In this study, we conducted QTL analyses for SL and SW using 186 recombinant inbred lines (RILs) derived from a cross between S1, an EMS mutant with extremely long siliques and large seeds, and S2, an inbred line with regular silique length and seed size. RILs were grown in Wuhan in the 2008/09 (SS09) and 2009/10 (SS10) growing seasons, and mean SL and SW for each line were investigated. Ten non-redundant QTL were identified for SL. Of these, a major QTL, cqSLA9, consistently explained as much as 53.4% of SL variation across environments. The others are minor QTL and individually explained less than 10% of the SL variation. Nine non-redundant QTL were identified for SW. Of which, one major QTL, cqSWA9, explained as much as 28.2% of the total SW variation in the SS09 and SS10 environments. In addition, three additive by additive interactions with small effects were detected for SL, and no interactions were detected for SW. Interestingly, the two major QTL, cqSLA9 for SL and cqSWA9 for SW colocalized in the same chromosomal region and were integrated into a unique QTL, uqA9. The S1 allele at this locus increases both SL and SW, suggesting that uqA9 has pleiotropic effects on both SL and SW. The existence and effect of uqA9 was confirmed in genetically different RILs derived from the cross between S1 and No2127, a resynthesized DH line having regular silique length and seed size. Individuals in one residual heterozygous line for cqSLA9 showed significant difference in silique length. The results in this study revealed that silique length in the S1 mutant is mainly controlled by the cqSLA9 locus, which will be suitable for fine mapping and marker-assisted selection in rapeseed breeding for high yield.     

QTL mapping of potato chip color and tuber traits within an autotetraploid family

Cultivated potato (Solanum tuberosum L.) is a highly heterozygous autotetraploid crop species, and this creates challenges for traditional line development and molecular breeding. Recent availability of a single-nucleotide polymorphism (SNP) array with 8303 features and software packages for linkage and association mapping in autotetraploid species present new opportunities for the identification of genomic regions that contribute to high-value traits in cultivated potato. A biparental tetraploid potato population was evaluated across three field seasons and storage trials in order to identify quantitative trait loci (QTL) for multiple tuber traits including fried chip color after 5.5–7.2 °C storage. Tetra-allelic dosage information was used to construct a genetic linkage map that covered 1041 cM and contained 2095 SNP markers with a median marker interval of 0.4 cM. A total of 41 QTL were identified for flower color, tuber yield, tuber number per plant, tuber weight, tuber size, and chip color after various storage regimes. Moderate effect QTL for chip color at 3 months were identified that co-localized with candidate genes vacuolar invertase (VInv), invertase inhibitor (INH2), and apoplastic invertase (Inv _ap -b). A separate QTL for chip color after 6 months of storage was identified in the short arm of chromosome 2, and this locus may contribute to variation in senescent sweetening resistance. QTL for tuber weight, length, and width co-localized with a known QTL for plant maturity on chromosome 5. Genome-wide association mapping using a polyploid model detected the tuber size QTL and identified a number of candidate SNPs, but was unable to detect markers significantly associated with chip color.     

Constructing high-density genetic maps for polyploid sugarcane (<Emphasis Type="Italic">Saccharum</Emphasis> spp.) and identifying quantitative trait loci controlling brown rust resistance

Sugarcane (Saccharum spp.) is an important economic crop for producing edible sugar and bioethanol. Brown rust has long been a major disease impacting sugarcane production worldwide. Resistance resource and markers linked to resistance are valuable tools for disease resistance improvement. An F₁ segregating population derived from a cross between two hybrid sugarcane clones, brown rust-susceptible CP95-1039 and brown rust-resistant CP88-1762, were genotyped using genotyping by sequencing approach and also phenotyped in a replicated field trial. Single nucleotide polymorphism (SNP) and presence/absence markers were called with seven different pipelines to maximize reliable marker identification. High-density maps were constructed for both parental clones with a total map length of 4224.4 cM, and a marker density of one marker per 1.7 cM for CP95-1039, and a total map length of 4373.2 cM, and one marker per 2.0 cM for CP88-1762. Among the seven SNP callers, Tassel and Genome Analysis ToolKit performed better than other callers in single-dose SNP detection and contribution to genetic maps. Two major quantitative trait loci (QTL) controlling brown rust resistance were identified, which can explain 21 and 30% of the phenotypic variation, respectively. The genetic maps generated here will improve our understanding of sugarcane’s complex genome structure and discovery of underlying sequence variations controlling agronomic traits. The putative QTL controlling brown rust resistance can effectively be utilized in sugarcane breeding programs to expedite the selection process of brown rust resistance after validation.     

Construction of a cucumber genetic linkage map with SRAP markers and location of the genes for lateral branch traits

Using SRAP (sequence-related amplified polymorphism) markers a genetic linkage map of cucumber was constructed with a population consisting of 138 F₂ individuals derived from a cross of the two cucumber lines, S06 and S52. In the survey of parental polymorphisms with 182 primer combinations, 64 polymorphism-revealing primer pairs were screened out, which generated totally 108 polymorphic bands with an average of 1.7 bands per primer pair and at most 6 bands from one primer pair. The constructed molecular linkage map included 92 loci, distributed in seven linkage groups and spanning 1164.2 cM in length with an average genetic distance of 12.6 cM between two neighboring loci. Based on this linkage map, the quantitative trait loci (QTL) for the lateral branch number (lbn) and the lateral branch average length (lbl) in cucumber were identified by QTLMapper1.6. A major QTL lbn1 located between ME11SA4B and ME5EM5 in LG2 could explain 10.63% of the total variation with its positively effecting allele from S06. A major QTL lbl1 located between DC1OD3 and DC1EM14 in LG2 could account for 10.38% of the total variation with its positively effecting allele from S₀₆.     

Restriction fragment length polymorphism analysis of loci associated with disease resistance genes and developmental traits in Pisum sativum L.

An F₂ population of pea (Pisum sativum L.) consisting of 174 plants was analysed by restriction fragment length polymorphism (RFLP) and random amplified polymorphic DNA (RAPD) techniques. Ascochyta pisi race C resistance, plant height, flowering earliness and number of nodes were measured in order to map the genes responsible for their variation. We have constructed a partial linkage map including 3 morphological character genes, 4 disease resistance genes, 56 RFLP loci, 4 microsatellite loci and 2 RAPD loci. Molecular markers linked to each resistance gene were found: Fusarium wilt (6 cM from Fw), powdery mildew (11 cM from er) and pea common Mosaic virus (15 cM from mo). QTLs (quantitative traits loci) for Ascochyta pisi race C resistance were mapped, with most of the variation explained by only three chromosomal regions. The QTL with the largest effect, on chromosome 4, was also mapped using a qualitative, Mendelian approach. Another QTL displayed a transgressive segregation, i.e. the parental line that was susceptible to Ascochyta blight had a resistance allele at this QTL. Analysis of correlations between developmental traits in terms of QTL effects and positions suggested a common genetic control of the number of nodes and earliness, and a loose relationship between these traits and height.     

SNP markers‐based map construction and genome‐wide linkage analysis in Brassica napus

An Illumina Infinium array comprising 5306 single nucleotide polymorphism (SNP) markers was used to genotype 175 individuals of a doubled haploid population derived from a cross between Skipton and Ag‐Spectrum, two Australian cultivars of rapeseed (Brassica napus L.). A genetic linkage map based on 613 SNP and 228 non‐SNP (DArT, SSR, SRAP and candidate gene markers) covering 2514.8 cM was constructed and further utilized to identify loci associated with flowering time and resistance to blackleg, a disease caused by the fungus Leptosphaeria maculans. Comparison between genetic map positions of SNP markers and the sequenced Brassica rapa (A) and Brassica oleracea (C) genome scaffolds showed several genomic rearrangements in the B. napus genome. A major locus controlling resistance to L. maculans was identified at both seedling and adult plant stages on chromosome A07. QTL analyses revealed that up to 40.2% of genetic variation for flowering time was accounted for by loci having quantitative effects. Comparative mapping showed Arabidopsis and Brassica flowering genes such as Phytochrome A/D, Flowering Locus C and agamous‐Like MADS box gene AGL1 map within marker intervals associated with flowering time in a DH population from Skipton/Ag‐Spectrum. Genomic regions associated with flowering time and resistance to L. maculans had several SNP markers mapped within 10 cM. Our results suggest that SNP markers will be suitable for various applications such as trait introgression, comparative mapping and high‐resolution mapping of loci in B. napus.