'百农64'×'京双16'小麦遗传连锁图谱构建

通过对小麦品种‘百农64’ב京双16’F3家系群体的SSR和AFLP分析,构建了含100个SSR标记(91个引物)和58个AFLP标记(12个引物)的小麦遗传连锁图,158个标记组成20个连锁群,覆盖小麦基因组3 114cM,标记间平均间距为19.7 cM.将前人未定位的12个SSR标记定位到了小麦遗传连锁图谱上.为小麦慢白粉病性等农艺性状的QTL分析打下了良好基础.     

梨分子遗传图谱构建及生长性状的QTL分析

利用鸭梨和京白梨杂交得到的F1(145株)实生苗为作图群体,通过对AFLP和SSR两种分子标记的遗传连锁分析,应用Joinmap 3.0作图软件,368个AFLP标记、34个SSR标记构建了分属18个连锁群的梨分子遗传连锁图谱,各连锁群的LOD值在4.0～7.0范围之间,图谱总长度覆盖梨基因组1395.9cM,平均图距为3.8cM.采用区间作图法,对该群体与生长性状相关的调查数据进行QTL分析,检测到与新梢生长量、新梢茎粗、节间长度、节间数量、树干径、树高及皮孔密度7个农艺性状连锁的QTL位点35个,其中主效QTL位点11个(LOD≥3.5).与生长性状相关的农艺性状QTL位点多集中在LG16连锁群上.     

小麦成株期白粉病抗性位点的QTL定位

基于高感白粉病小麦品种宁春4号和高抗白粉病品种宁春27号构建的重组自交系群体(RIL),以及简化基因组测序方法获得的遗传连锁图谱,通过QTL定位分析了小麦成株期白粉病抗性基因.结果 表明:4对主基因控制的加性-上位性遗传模型适合于RIL群体的成株期白粉病抗性;2个稳定的抗病QTL QPm.naafs-4D和QPm.na...     

南瓜遗传连锁图谱的构建及粒宽性状的QTL定位

以印度南瓜纯系大粒材料‘0515-1’和小粒材料‘0460-1-1’为亲本,获得193个南瓜F2单株群体,应用AFLP和SSR分子标记技术进行多态性筛选,构建了含84个标记位点的遗传连锁图谱。结果表明,整个图谱包含12个连锁群,全长683.50cM,标记平均间距为8.13cM。采用复合区间定位分析,共检测到控制南瓜籽粒宽度的4个数量性状位点(QTL),分别位于3个连锁群上,各QTL的贡献率在2.87%～29.68%之间。     

小麦全基因组抗赤霉病QTL关联位点特异性SSR标记的筛选、等位变异及效应解析

赤霉病是小麦主要的流行病害之一。借助标记辅助选择将不同数量性状基因座（quantitative trait loci，QTL）聚合是防治赤霉病有效且环保的方法，可以从源头上控制赤霉病并降低籽粒中毒素含量。抗赤霉病QTL在小麦全基因组均有分布，但除了Fhb1、Fhb2等少数位点有比较可靠的鉴别标记，绝大部分位点缺乏有效的位点特异性鉴别标记。简单重复序列（simple sequence repeat，SSR）标记多态性丰富，可以区分自然群体中不同等位变异，方便用于标记辅助育种。基于此，搜集了不同文献中报道的与赤霉病关联的SSR标记386个，并用这些标记构建全基因组赤霉病抗性QTL一致性图谱，接着对这些关联标记进行拷贝数分析，进而选择位点内的单拷贝SSR标记，将这些单拷贝标记在156个品种组成的自然群体中进行扩增，并与三季大田和三季温室环境下赤霉病抗性进行关联，筛选与赤霉病抗性关联的单拷贝SSR标记，明确这些标记在自然群体中的有效等位变异和效应。结果表明，共8个单拷贝SSR标记至少在两季试验中与表型显著关联(P<0.05)，涉及2B、2D、3B、5A、5B、6A、6D、7A染色体，有5个单拷贝标记位点存在有效等位变异。中国地方品种和日本品种携带更多的有利变异，且有利等位变异数目越多的品种赤霉病抗性越好。研究分析的QTL位点及其关联的单拷贝SSR标记可用于赤霉病抗病育种，有利于提高品种赤霉病抗性水平和育种效率。     

基于SRAP和SSR标记构建的杏扁遗传连锁图谱

以杏扁品种‘龙王帽’授粉‘优一’获得98个F1代单株为作图群体,采用SRAP和SSR标记进行连锁图谱的构建。采用Join Map 4.0软件进行连锁分析,分别构建了‘龙王帽’和‘优一’的分子连锁框架图,共获得132个SRAP标记和17个SSR标记。其中父本遗传图谱涉及8个连锁群,包含53个SRAP标记和9个SSR标记,图谱总长为694.8 cM,标记间平均图距为11.21 cM,平均每个连锁群上有7.75个标记位点,连锁群平均长度为86.85 cM;母本遗传图谱涉及8个连锁群,包含79个SRAP标记和8个SSR标记,图谱总长为924.8 cM,标记间平均图距为10.63 cM,平均每个连锁群上有10.87个标记位点,连锁群平均长度为115.6 cM。     

大豆昆虫抗性相关QTLs的元分析

大豆虫害严重危害大豆生产。虽然大豆抗虫相关QTLs研究增多, 但由于作图群体不同、同种昆虫抗性QTL的调查性状不同以及数据分析方法存在差异等原因, 使QTL精确性和有效性被降低。因此, 获得相对真实且有效的QTLs位点对于促进分子标记辅助选择有重要意义。文章通过搜集已报道的81个与大豆昆虫抗性相关的QTL, 提取相对有效且可靠的QTLs标记信息, 利用元分析软件BioMercator2.1将这些QTLs映射到大豆公共遗传连锁图谱Soymap2上, 通过单独与联合的两种元分析途径, 利用QTLs的95%的置信区间来推断“真实QTLs”的位置。文章不仅构建了一张大豆昆虫抗性一致性图谱, 而且通过两种元分析途径分别得到12个和14个QTLs位点, 且其中有6个位点QTL的位置一致。它们被定位在9个连锁群上, 主要成簇分布在E、F、H、M等4个连锁群上, 图距由原来平均15 cM缩减到平均3.67 cM。除了一个与大豆食心虫抗性相关的位点外, 其余QTLs都与多种昆虫抗性相关。研究结果明显缩短了原来已报道的QTL置信区间, 为大豆抗虫相关QTL的精细定位以及抗虫相关基因挖掘提供了依据。     

人工合成小麦抗白粉病未知基因的SSR标记

YAV-2/TEZ//A.SQ(895)是硬粒小麦与粗山羊草杂交获得的抗白粉病人工合成小麦。本研究利用人工合成小麦YAV-2/TEZ//A.SQ(895)与感白粉病的普通小麦品系品资50098杂交和自交获得的F2代群体及F3家系,在温室条件下鉴定群体的白粉病抗性。遗传分析结果表明,该抗白粉病基因为显性单基因遗传。利用647对小麦SSR引物进行了白粉病抗性基因的分子标记分析,结果表明该白粉病抗性基因与2A染色体的6个SSR标记连锁,与标记Xcfa2086的遗传距离最近,为11.8cM。     

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。     

应用重组自交系群体定位大豆抗虫QTL

以大豆组合科丰1号×南农1138-2衍生的重组自交系(RIL)群体为材料构建遗传连锁图谱, 利用软件 Cartographer V.2.5 采用复合区间作图法检测定位大豆抗虫QTL。以斜纹夜蛾幼虫重为抗性指标, 检测到 1 个与抗虫性有关的 QTL, 位于G20-O连锁群上, 其端距离为31.91 cM, 加性效应估计值为0.0408, 对性状变异的解释率为 11.74％; 以蛹重为抗性指标, 检测到 2 个与抗虫性有关的 QTL, 分别位于G8-D1b+W和G17-L连锁群上, 其端距离分别为 14.71 cM和0.01 cM, 加性效应估计值分别为-0.0139和0.0103, 对性状变异的解释率分别为 11.30％和6.36％。     

A major QTL controlling seed dormancy and pre-harvest sprouting resistance on chromosome 4A in a Chinese wheat landrace

Wheat pre-harvest sprouting (PHS) can cause significant reduction in yield and end-use quality of wheat grains in many wheat-growing areas worldwide. To identify a quantitative trait locus (QTL) for PHS resistance in wheat, seed dormancy and sprouting of matured spikes were investigated in a population of 162 recombinant inbred lines (RILs) derived from a cross between the white PHS-resistant Chinese landrace Totoumai A and the white PHS-susceptible cultivar Siyang 936. Following screening of 1,125 SSR primers, 236 were found to be polymorphic between parents, and were used to screen the mapping population. Both seed dormancy and PHS of matured spikes were evaluated by the percentage of germinated kernels under controlled moist conditions. Twelve SSR markers associated with both PHS and seed dormancy were located on the long arm of chromosome 4A. One QTL for both seed dormancy and PHS resistance was detected on chromosome 4AL. Two SSR markers, Xbarc 170 and Xgwm 397, are 9.14 cM apart, and flanked the QTL that explained 28.3% of the phenotypic variation for seed dormancy and 30.6% for PHS resistance. This QTL most likely contributed to both long seed dormancy period and enhanced PHS resistance. Therefore, this QTL is most likely responsible for both seed dormancy and PHS resistance. The SSR markers linked to the QTL can be used for marker-assisted selection of PHS-resistant white wheat cultivars. Shi-Bin Cai and Cui-Xia Chen contributed equally to this work.     

A new intervarietal linkage map and its application for quantitative trait locus analysis of "gigas" features in bread wheat.

A doubled-haploid (DH) population from an intervarietal cross between the Japanese cultivar 'Fukuho-komugi' and the Israeli wheat line 'Oligoculm' was produced by means of wheat x maize crosses. One hundred seven DH lines were genotyped to construct a simple sequence repeat (SSR) based linkage map with RFLP, RAPD, and inter-simple sequence repeat markers. Out of 570 loci genotyped, 330 were chosen based on their positions on the linkage map to create a "framework" map for quantitative trait locus (QTL) analysis. Among the 28 linkage groups identified, 25 were assigned to the 21 chromosomes of wheat. The total map length was 3948 cM, including the three unassigned linkage groups (88 cM), and the mean interval between loci was 12.0 cM. Loci with segregation distortion were clustered on chromosomes 1A, 4B, 4D, 5A, 6A, 6B, and 6D. After vernalization, the DH lines were evaluated for spike number per plant (SN) and spike length (SL) in a greenhouse under 24-h daylength to assess the "gigas" features (extremely large spikes and leaves) of 'Oligoculm'. The DH lines were also autumn-sown in the field in two seasons (1990-1991 and 1997-1998) for SN and SL evaluation. QTL analysis was performed by composite interval mapping (CIM) with the framework map to detect QTLs for SN and SL. A major QTL on 1AS, which was stable in both greenhouse and field conditions, was found to control SN. This QTL was close to the glume pubescence locus (Hg) and explained up to 62.9% of the total phenotypic variation. The 'Oligoculm' allele restricted spike number. The SSR locus Xpsp2999 was the closest locus to this QTL and is considered to be a possible marker for restricted tillering derived from 'Oligoculm'. Eight QTLs were detected for SL. The largest QTL detected on 2DS was common to the greenhouse and field environments. It explained up to 33.3% of the total phenotypic variation. The second largest QTL on 1AS was common to the greenhouse and the 1997-1998 season. The position of this QTL was close to that for the SN detected on 1AS. The association between SN and SL is discussed.     

An integrated DArT-SSR linkage map of durum wheat

Genetic mapping in durum wheat (Triticum durum Desf.) is constrained by its large genome and allopolyploid nature. We developed a Diversity Arrays Technology (DArT) platform for durum wheat to enable efficient and cost-effective mapping and molecular breeding applications. Genomic representations from 56 durum accessions were used to assemble a DArT genotyping microarray. Microsatellite (SSR) and DArT markers were mapped on a durum wheat recombinant inbred population (176 lines). The integrated DArT-SSR map included 554 loci (162 SSRs and 392 DArT markers) and spanned 2022 cM (5 cM/marker on average). The DArT markers from durum wheat were positioned in respect to anchor SSRs and hexaploid wheat DArT markers. DArT markers compared favourably to SSRs to evaluate genetic relationships among the durum panel, with 1315 DArT polymorphisms found across the accessions. Combining DArT and SSR platforms provides an efficient and rapid method of generating linkage maps in durum wheat.     

Molecular tagging of a rust resistance gene in cultivated groundnut (Arachis hypogaea L.) introgressed from Arachis cardenasii

Rust is a serious and the most prevalent groundnut disease in tropical and subtropical growing regions of the world. A total of 164 recombinant inbred lines derived from resistant (VG 9514) and susceptible (TAG 24) cultivated groundnut parents were screened for rust resistance in five environments. Subsequent genotyping of these lines with 109 simple sequence repeat (SSR) markers generated a genetic linkage map with 24 linkage groups. The total length of the linkage map was 882.9 cM with an average of 9.0 cM between neighbouring markers. The markers pPGPseq4A05 and gi56931710 flanked the rust resistance gene at map distances of 4.7 cM and 4.3 cM, respectively, in linkage group 2. The significant association of these two markers with the rust reaction was also confirmed by discriminant analysis. The informative SSR markers classified rust-resistant and susceptible groups with 99.97% correctness. The SSR markers pPGPseq4A05 and gi56931710 were able to identify all the susceptible genotypes from a set of 20 cultivated genotypes differing in rust reaction. Tagging of the rust resistance locus with linked SSR markers will be useful in selecting the rust resistant genotypes from segregating populations and in introgressing the rust resistance genes from diploid wild species.     

Identification of flanking SSR markers for a major rice gall midge resistance gene <Emphasis Type="Italic">Gm1</Emphasis> and their validation

Host-plant resistance is the preferred strategy for management of Asian rice gall midge (Orseolia oryzae), a serious pest in many rice-growing countries. The deployment of molecular markers linked to gall midge resistance genes in breeding programmes can accelerate the development of resistant cultivars. In the present study, we have tagged and mapped a dominant gall midge resistance gene, Gm1, from the Oryza sativa cv. W1263 on chromosome 9, using SSR markers. A progeny-tested F₂ mapping population derived from the cross W1263/TN1 was used for analysis. To map the gene locus, initially a subset of the F₂ mapping population consisting of 20 homozygous resistant and susceptible lines each was screened with 63 parental polymorphic SSR markers. The SSR markers RM316, RM444 and RM219, located on chromosome 9, are linked to Gm1 at genetic distances of 8.0, 4.9 and 5.9 cM, respectively, and flank the gene locus. Further, gene/marker order was also determined. The utility of the co-segregating SSR markers was tested in a backcross population derived from the cross Swarna/W1263//Swarna, and allelic profiles of these markers were analysed in a set of donor rice genotypes possessing Gm1 and in a few gall midge-susceptible, elite rice varieties.     

Saturation and mapping of a majorFusarium head blight resistance QTL on chromosome 3BS of Sumai 3 wheat

Fusarium head blight (FHB) is a destructive disease in wheat. The major quantitative trait locus (QTL) on 3BS from Sumai 3 and its derivatives has been used as a major source of the resistance to FHB worldwide, but the discrepancy in reported location of the major QTL could block its using in map based cloning and marker assisted selection. In this study, Chinese Spring-Sumai 3 chromosome 3B substitution line was used as resistant parent of the mapping population to reduce the confounded effect of genetic background in Sumai 3. The major QTL region was saturated with the Sequence Tagged Microsatellite (STM) and Sequence Tagged Site (STS) markers. A linkage map of chromosome 3B with 36 markers covering a genetic distance of 112.4 cM was constructed. Twelve new markers were inserted into the chromosome region where the major QTL was located. The average interval distance between markers was 1.5 cM. Multiple QTL Models (MQM) mapping indicated that the major QTL was located in the interval ofXgwm533 — Xsts9-1, and explained 45.6% of phenotypic variation of the resistance to FHB. The SSR (simple sequence repeat) markerXgwm533 and STM markerXstm748tcac are closely linked to the major QTL.     

QTL analysis for resistance to bacterial wilt (Burkholderia caryophylli) in carnation (Dianthus caryophyllus) using an SSR-based genetic linkage map

Bacterial wilt (Burkholderia caryophylli (Burkholder) Yabuuchi et al.) is one of the most damaging diseases during carnation (Dianthus caryophyllus L.) cultivation in Japan. To find molecular markers for use in marker-assisted selection, we constructed a simple sequence repeat (SSR)-based genetic linkage map of carnation using an F₂ population of 90 plants derived from a cross between a highly resistant line (85-11) and a susceptible cultivar (Pretty Favvare). To develop a large number of SSR markers, we constructed four new SSR-enriched genomic libraries and conducted expressed sequence tag analysis. We mapped 178 SSR loci into 16 linkage groups. The map covered 843.6?cM, with an average distance of 6.5?cM between two loci. This is the first report of a genetic linkage map based mainly on SSR markers in the genus Dianthus. Quantitative trait locus (QTL) analysis identified one locus for resistance to bacterial wilt in linkage group (LG) B4. The locus explained 63.0% of the phenotypic variance for resistance to bacterial wilt. The SSR markers CES1161 and CES2643 that were closest to the QTL were efficient markers for selecting lines with resistance derived from line 85-11. A positional comparison using SSR markers as anchor loci revealed that LG B4 corresponded to LG A6 in a previously constructed map. We found that the position of the resistance locus derived from line 85-11 was similar to that of the major resistance locus observed for a highly resistant wild species, Dianthus capitatus ssp. andrzejowskianus.     

Integration genetic linkage map construction and several potential QTLs mapping of Chinese shrimp (Fenneropenaeus chinensis) based on three types of molecular markers

In this study, totally 54 selected polymorphic SSR loci of Chinese shrimp (Fenneropenaeus chinensis), in addition with the previous linkage map of AFLP and RAPD markers, were used in consolidated linkage maps that composed of SSR, AFLP and RAPD markers of female and male construction, respectively. The female linkage map contained 236 segregating markers, which were linked in 44 linkage groups, and the genome coverage was 63.98%. The male linkage map contained 255 segregating markers, which were linked in 50 linkage groups, covering 63.40% of F. chinensis genome. There were nine economically important traits and phenotype characters of F. chinensis were involved in QTL mapping using multiple-QTL mapping strategy. Five potential QTLs associated with standard length (q-standardl-01), with cephalothorax length (q-cephal-01), with cephaloghorax width (q-cephaw-01), with the first segment length (q-firsel-01) and with anti-WSSV (q-antiWSSV-01) were detected on female LG1 and male LG44 respectively with LOD> 2.5. The QTL q-firsel-01 was at 73.603 cM of female LG1. Q-antiWSSV-01 was at 0 cM of male LG44. The variance explained of these five QTLs was from 19.7-33.5% and additive value was from -15.9175 to 7.3675. The closest markers to these QTL were all SSR, which suggested SSR marker was superior to AFLP and RAPD in the QTL mapping.     

Construction of genetic linkage maps and QTL mapping for X-disease resistance in tetraploid chokecherry (Prunus virginiana L.) using SSR and AFLP markers

Chokecherry (Prunus virginiana L.) (2n = 4x = 32) is a unique Prunus species for both genetics and disease resistance research due to its tetraploid nature and known variations in X-disease resistance. X-disease is a destructive disease of stone fruit trees, causing yield loss and poor fruit quality. However, genetic and genomic information on chokecherry is limited. In this study, simple sequence repeat (SSR) and amplified fragment length polymorphism (AFLP) markers were used to construct genetic linkage maps and to identify quantitative trait loci (QTLs) associated with X-disease resistance in chokecherry. A segregating population (101 progenies) was developed by crossing an X-disease-resistant chokecherry line (RC) with a susceptible chokecherry line (SC). A total of 498 DNA markers (257 SSR and 241 AFLP markers) were mapped on the two genetic maps of the two parental lines (RC and SC). The map of RC contains 302 markers assigned to 14 linkage groups covering 2,089 cM of the genome. The map of SC has 259 markers assigned to 16 linkage groups covering 1,562.4 cM of the genome. The average distance between two markers was 6.9 cM for the RC map and 6.0 cM for the SC map. One QTL located on linkage group 15 on the map of SC was found to be associated with X-disease resistance. Genetic linkage maps and the identified QTL linked to X-disease resistance will further facilitate genetic research and breeding of X-disease resistance in chokecherry and other Prunus species.     

Genetic mapping and localization of a major QTL for seedling resistance to downy mildew in Chinese cabbage (Brassica rapa ssp. pekinensis)

Downy mildew caused by the fungus Peronospora parisitica is a serious threat to members of the Brassicaceae family. Annually, a substantial loss of yield is caused by the widespread presence of this disease in warm and humid climates. The aim of this study was to localize the genetic factors affecting downy mildew resistance in Chinese cabbage (Brassica rapa ssp. pekinensis). To achieve this goal, we improved a preexisting genetic map of a doubled-haploid population derived from a cross between two diverse Chinese cabbage lines, 91-112 and T12-19, via microspore culture. Microsatellite simple sequence repeat (SSR) markers, isozyme markers, sequence-related amplified polymorphism markers, sequence-characterized amplified region markers and sequence-tagged-site markers were integrated into the previously published map to construct a composite Chinese cabbage map. In this way, the identities of linkage groups corresponding to the Brassica A genome reference map were established. The new map contains 519 markers and covers a total length of 1,070 cM, with an average distance between markers of 2.06 cM. All markers were designated as A1–A10 through alignment and orientation using 55 markers anchored to previously published B. rapa or B. napus reference maps. Of the 89 SSR markers mapped, 15 were newly developed from express sequence tags in Genbank. The phenotypic assay indicated that a single major gene controls seedling resistance to downy mildew, and that a major QTL was detected on linkage group A8 by both interval and MQM mapping methods. The RAPD marker K14-1030 and isozyme marker PGM flanked this major QTL in a region spanning 2.9 cM, and the SSR marker Ol12G04 was linked to this QTL by a distance of 4.36 cM. This study identified a potential chromosomal segment and tightly linked markers for use in marker-assisted selection to improve downy mildew resistance in Chinese cabbage.