大豆连锁群Ⅰ中种子蛋白质QTL的精细图谱及其对农艺性状的相关效应

大豆是一种重要的蛋白质和油分资源。以前将控制种子蛋白质含量的QTL绘制在大豆连锁群Ⅰ上。本文旨在对该QTL进行精细作图，并弄清是否有另外的重组体能减少种子蛋白质含量和产量和含油量问负向表型相关。采用两组回交群体构建精细图谱。这些群体是将野生大豆（G．soja）P1468916的高蛋白质等位基因导人A81—356022育种品系的遗传背景构建而成。第一组群体包含BC5品系的3个群体，第二组群体包含BC，品系的4个群体。这些群体QTL图谱基因组区域的不同片断有分离。两组群体的测试结果将控制蛋白质和含油量的QTL定位在SSR标记Satt239和AFLP标记ACG9b之间的3cM间隔处。农艺性状评价试验结果的不一致性使我们很难确切地推断蛋白质QTL是否通过多效性控制了其它性状。     

甘蓝型油菜籽粒含油量、蛋白质、纤维素及半纤维素含量QTL分析

以甘蓝型黄籽油菜GH06和甘蓝型黑籽油菜P174为亲本,通过单粒法连续自交8代构建重组自交系群体,应用SSR标记绘制31个连锁群(LGs)的遗传连锁图谱,图谱总长1437.1 cM,相邻标记间的平均距离为3.89 cM。对4个不同环境下RIL8群体中每个株系籽粒含油量、蛋白质、纤维素和半纤维素含量进行了近红外分析,性状相关性表明含油量与其他3个性状均表现负相关,蛋白质含量与纤维素和半纤维素分别表现负相关和正相关。结合构建的遗传图谱采用复合区间作图法分析4个性状QTL,共检测到26个QTL,分布在N2、N3、N8、N9、N11、N13、N16和N17连锁群上,其中8个含油量QTL可解释表型变异的4.96%~21.83%;6个蛋白含量QTL,可解释表型变异的3.12%~14.28%;4个纤维素含量QTL,可解释表型变异的4.60%~17.29%;8个半纤维素含量QTL,可解释表型变异率的6.66%~16.68%。在N8上,发现有含油量QTL与半纤维素含量QTL重叠的区段。在N9上,发现有纤维素含量QTL与半纤维素含量QTL重叠的区段,上述2个区段重叠QTL加性效应方向相反。本研究认为油菜种子含油量、蛋白质、纤维素和半纤维素属于典型的数量性状,受环境影响较大,与这些QTL紧密相关的分子标记可为下一步分子标记辅助育种提供一定技术支撑。     

黄瓜种子长度的遗传分析及分子标记

为了今后正确制定籽用黄瓜育种方案和有效地进行品种选育,有必要对黄瓜种子大小进行详细研究。选用种子长短不同的5个黄瓜自交系作为杂交亲本,按照Griffing双列杂交试验方法Ⅱ配制杂交组合。以长种子自交系D06157和短种子自交系D0603为亲本,构建F2分离群体,采用SSR分子标记构建遗传连锁图谱,利用复合区间定位方法进行QTL定位。结果表明：黄瓜种子长度属于数量性状,以显性效应为主,同时存在加性效应,广义遗传力及狭义遗传力较低,分别为28.6777%、10.8384%。构建的4个连锁群含有16个SSR位点,连锁群总长度为287.1 cM,标记间平均距离为17.94 cM。检测出6个与黄瓜种子长度相关的QTL,其中QTL5和QTL6有着较高的对种子长度性状变异的解释率(>5%),距离最近标记的图距分别为2.1 cM、4.3 cM,LOD值分别为7.84、9.69,可解释遗传变异分别为6.07%、6.08%,加性效应值分别为39.12%、37.21%。     

大白菜紫色性状的SRAP连锁标记的筛选

以紫色大白菜BC1回交群体的120个单株为材料,采用混合集群分析法构建大白菜的紫色与非紫色池,应用SRAP分子标记技术筛选两池间的多态性,并对与紫色性状相关的QTL和对应的连锁关系进行了初步分析.结果表明,在144对SRAP引物组合中,仅有8对引物组合可以在两池间扩增出较稳定的多态性产物.应用获得的8对引物组合对120个单株的DNA进行SRAP分析,初步获得3个与紫色性状相关的QTL:qp-c1-1、qp-c1-2和qp-c1-3,同时获得6个与这3个QTL连锁的标记,其中m7e11-490、m6e8-210、m7e11-900n和m6e8-100n 4个标记与其最近QTL位点的连锁距离分别为0.1 cM、0.1 cM、0.2 cM和2.2 cM.这为紫色白菜的分子辅助育种提供了科学依据.     

山农01-35×藁城9411重组自交系遗传图谱构建及粒重QTL分析

为利用分子遗传图谱进行小麦产量数量性状位点定位分析,以大粒高产小麦品系山农01-35和强筋小麦藁城9411为亲本,衍生了含182个家系的重组自交系(RIL)F₈群体,用442个DArT标记、59个SSR标记和1个TaGW2-CAPS标记,构建了包括29个连锁群的分子遗传图谱,总遗传长度为4 084.5 cM,标记间平均距离为8.13 cM。定位了54个新标记位点,包括44个DArT和10个SSR标记,分布于除4D、6B、7B外的其他18条染色体上。用该分子遗传图谱和4个环境粒重表型值,共检测到7个影响粒重的加性QTL,分别位于1B、4B、5B、6A染色体,其中QGW4B-7、QGW5B-20和QGW6A-29在单环境分别定位和4个环境共同定位两种方法中均能检测到。QGW4B-7、QGW5B-12和QGW6A-29对粒重的贡献率均超过10%,为主效QTL。本研究结果可为小麦高产优质性状的QTL分析及分子标记辅助选择提供参考。     

花生InDel标记开发及其在含油量QTL定位中的应用

培育高含油量品种是花生重要的育种目标,开发稳定高效的分子标记对于标记辅助选择花生高含油量品种具有重要价值。本研究针对2个含油量差异显著的亲本徐花13 (高含油量)和中花6号(低含油量),利用PacBio三代测序技术检测基因组结构变异,分别从徐花13和中花6号中检测到35,794个和74,703个结构变异。根据双亲结构变异信息,针对前期定位的含油量区间开发InDel (插入/缺失)标记84个,其中9个InDel标记的扩增片段证实在双亲间具有多态性。基于RIL (重组自交系)群体中的杂合残余获得的NIL (近等基因系)构建的一个包含1160个单株的F2群体,使用9个多态性InDel标记鉴定其基因型,构建了总长度为149.84 cM的局部遗传连锁图谱。结合F2群体的含油量表型数据,将该含油量QTL定位在标记M23至M11之间,位于A08染色体1.2 Mb区段内。本试验证实了InDel标记开展花生含油量QTL定位的可行性,其定位的含油量位点及其紧密连锁的InDel标记能够为花生分子标记辅助育种提供理论和技术指导。     

应用SSR标记构建高油玉米籽粒含油量的QTL图谱

高油玉米越来越受到人们的关注。本试验研究了北京高油种质（BHO）籽粒含油量的QTL及其遗传效应。我们将源于高油群体（BHO Cycle 13）的高油自交系By804与一个重要常规自交系杂交，构建了包含450个个体的F2群体。用核磁共振（NMR）测定F2或F3种子的单粒含油量。选用150个共显性SSR标记构建长1759．1cM、平均问隔11．65cM的遗传连锁图。共构建了20个与籽粒含油量有关的QTL图谱，LOD临介值为3．54。在F2和F，种子中同时检测到的有6个QTL，占总图谱QTL的30％，而另外14个QTL则是在单个子代中检测出的。单个QTL占表型变异比例的幅度为2．31—17．51％。     

利用海岛棉染色体片段导入系定位衣分和籽指QTL

以染色体片段导入系IL-15-5和IL-15-5-1构建的F2和F2:3分离群体,利用SSR标记对数量性状衣分和籽指QTL进行了定位。应用复合区间作图法分析两个组合的774个F2单株和F2:3家系衣分和籽指,检测到2个衣分的QTL,1个籽指的QTL。衣分QTLqLP-15-1在两世代中都被检测到,位于相同的分子标记置信区间JESPR152～NAU3040,置信的遗传距离分别为5.40cM和3.20cM;qLP-15-2只在F2:3中被检测到,位于分子标记NAU5302～NAU2901之间,置信的遗传距离为0.08cM。籽指QTLqSI-15-1在F2和F2:3中都被检测到,分别位于分子标记NAU2814～NAU3040和JESPR152～NAU3040,置信的遗传距离分别为6.70cM和5.70cM。利用染色体片段导入系能准确地定位产量组分的QTL,为棉花产量的分子设计育种奠定基础。     

玉米种子休眠性的QTL定位

选用两个种子休眠性差异较大的普通玉米自交系R08与A318组配的F2群体共331个单株,构建了包含137个SSR标记的分子遗传连锁图谱,覆盖玉米基因组2 076.7 cM,平均图距15.2 cM。采用复合区间作图法对F_2:3家系种子休眠性数据进行分析,共检测到7个QTL,分别位于玉米第1、3、5和10染色体上。7个QTL的贡献率在2.45%~26.     

大豆重组自交系群体NJRIKY遗传图谱的加密及其应用效果

作物基因组研究,包括基因或数量性状位点(QTL)定位、图位克隆以及物理图谱构建等,首先必须建立具有丰富标记信息的高密度遗传连锁图谱。由科丰1号和南农1138-2杂交组合衍生的重组自交系群体NJRIKY已经构建了4张大豆遗传连锁图谱,但由于遗传信息和标记数目不够充分,在基因和QTL作图时仍然存在精确度和准确度问题。为增加NJRIKY图谱密度,本研究在967对SSR引物中获得了401个多态性SSR标记。结合其他分子数据,使用作图软件Mapmaker/Exp3.0b,获得一张含有553个遗传标记,25个连锁群,总长2071.6cM,平均图距3.70cM的新遗传连锁图谱,其中SSR标记316个,RFLP标记197个,EST标记39个,形态标记1个。连锁群上大于20cM的标记间隔由原来42个减少到2个。原图谱的3个SMV抗性基因定位于D1b连锁群末端的开放区间上且仅与一个RFLP标记连锁,利用加密图谱对Rsc-3、Rsc-7、Rsc-9、Rsc-13、Rsa、Rn1和Rn3等7个SMV抗性基因重定位,全部位于D1b连锁群,与相邻分子标记距离均小于6cM,其中Rsc-9、Rn1、Rsa的距离小于1cM,Rsc-13与EST标记GMKF168a共分离。对本群体农艺性状进行QTL重定位,获得8个性状相关的42个主效QTL,其中20个QTL遗传贡献率大于10%,与原图谱比较,新定位的各QTL的标记区间明显缩短,与相邻标记的连锁更加紧密。     

崇化大梨×新世纪梨F1代分子遗传图谱的构建及两个果实性状的QTL分析

利用崇化大梨×新世纪梨杂交得到的94株F1代实生苗为作图群体,应用mapmaker/exp 3.0软件构建了一张包括19个SSR标记,315个SRAP标记,合计335个标记分属于18个连锁群的遗传图谱,图谱总长度为1 300 cM,平均图距为3.9 cM。采用区间作图法检测到与果实发育期、果形指数两个果实性状相连锁的QTL位点7个,其中主效的QTL位点2个(LOD≥3.5),分别位于LG2和LG17连锁群。     

Dissection of genetic architecture for oil content in soybean seed using two backcross populations

Soybean seed oil was valued in foods, animal feed and some industrial applications. Molecular marker‐assisted selection (MAS) for high‐oil‐content cultivars was an important method for soybean breeders. The objective of this study was to identify quantitative trait loci (QTL) and epistatic QTL underlying the seed oil content of soybeans across two backcross (BC) populations (with one common male parent ‘Dongnong47’) and two different environments. Two molecular genetic maps were constructed. They encompassed 1046.8 cM [with an average distance of 6.75 cM in the ‘Dongnong47’  ×  ‘Jiyu89’ (DJ) population] and 846.10 cM [with an average distance of 5.76 cM in the ‘Dongnong47’  ×  ‘Zaoshu18’ (DZ) population]. Nine and seven QTL were identified to be associated with oil content in the DJ and DZ populations, respectively. The phenotypic variation explained by most of the QTL was usually less than 10%. Among the identified QTL, those stable ones across multiple environments and populations often had stronger additive effects. In addition, three stable QTL in the DZ populations were identified in the similar genomic region of the three QTL in the DJ population [qDJE and qDZE‐1 were located near Satt151 of Chromosome 15 (Chr15), qDJA1 and qDZA1 were located near Satt200 of Chr15 (LG A1), and qDJD2‐1 and qDZD2‐1 were located near Sat365 of Chr17]. In conclusion, MAS will be able more effectively to combine beneficial alleles of the different donors to design new genotypes with higher soybean seed oil content using the BC populations.     

白芝麻籽粒油脂、蛋白质及芝麻素含量QTL定位分析

芝麻籽粒油脂、蛋白质和芝麻素含量是芝麻品质育种的3个重要目标。为了解析其遗传机制并检测相关QTL,利用近红外谷物品质分析仪对一个包含224个株系的F9代重组自交系(RIL)群体在2年3个环境下的籽粒品质性状检测,结果表明,群体内株系间差异显著且呈典型正态分布,而同一环境不同重复间表现差异不显著。相关性分析显示,籽粒含油量与蛋白质含量显著负相关,籽粒含油量与芝麻素含量显著正相关,蛋白质含量与芝麻素含量显著负相关;利用该RIL群体已构建的高密度遗传图谱,采用基于混合线性模型的复合区间作图法(MCIM)检测到8个QTL,表型贡献率为0.41%~14.55%;采用多重区间作图法(MIM)检测到13个QTL,可解释5.2%~18.6%的表型变异。其中5个主效QTL被2种方法同时检测到且定位区间相同,2个主效QTL在2个或3个环境中被重复检测到。控制含油量的Qoc-5与芝麻素含量的Qsc-5位于LG5连锁群上的相同区段,加性效应均为正值;而控制蛋白质含量的Qpc-5也位于相邻位置,但加性效应为负值。LG2和LG1连锁群上也存在相似情况,反映品质性状相关QTL之间存在一因多效或紧密连锁。因此,在芝麻品质育种中选择高含油量可以兼顾高芝麻素,但应对蛋白质含量进行负向选择。     

甘蓝型油菜产量及相关性状的QTL分析

高产是甘蓝型油菜育种的重要目标之一，产量是多基因控制的数量性状。本文通过QTL作图分析了产量及其相关性状的数量性状位点，以甘蓝型油菜中油821和保604 F₁代小孢子培养获得的DH系为作图群体，构建了由20个连锁群组成的，包括251个分子标记( 2个RFLP标记，72个RAPD标记，91个SSR标记，86个SRAP标记)的遗传连锁图(10个标记没有分配到连锁群中)。图谱的平均图距为6.96 cM，共覆盖油菜基因组1 746.5 cM。在此图谱基础上采取复合区间作图法，检测到与油菜产量及其相关性状有关的QTL共17个。其中控制株高的3个分别位于第4、第9和第10连锁群上，对性状的解释率为9.42%～17.58%；与分枝部位有关的4个分别位于第4、第6和第7连锁群上，其中Bp1 和Bp2 均位于第4连锁群，对性状的解释率为8.13%～15.20%；与主花序有效长有关的3个分别位于第4、第10和第16连锁群上，对性状的解释率为7.49%～23.36%；与一次有效分枝有关的2个分别位于第1、第4连锁群上，对性状的解释率为15.29%～19.58%；与角果总数和千粒重有关的分别位于第4连锁群和第9连锁群上，贡献率分别为17.42%和7.64%；与单株产量有关的3个分别位于第3、第4和第15连锁群，共解释26.60%的表型变异。部分性状的QTL在连锁群上成簇分布,对性状贡献率很大，表现主效QTLs的特点，相应的性状之间也呈显著相关，这表明一因多效或者相关的QTLs之间紧密连锁是性状相关的遗传基础。本研究中与主效QTLs连锁的标记可用于油菜产量性状的分子标记辅助选择。     

Mapping and validation of QTLs for resistance to an Indian isolate of Ascochyta blight pathogen in chickpea

Ascochyta blight (AB) caused by Ascochyta rabiei, is globally the most important foliar disease that limits the productivity of chickpea (Cicer arietinum L.). An intraspecific linkage map of cultivated chickpea was constructed using an F₂ population derived from a cross between an AB susceptible parent ICC 4991 (Pb 7) and an AB resistant parent ICCV 04516. The resultant map consisted of 82 simple sequence repeat (SSR) markers and 2 expressed sequence tag (EST) markers covering 10 linkage groups, spanning a distance of 724.4 cM with an average marker density of 1 marker per 8.6 cM. Three quantitative trait loci (QTLs) were identified that contributed to resistance to an Indian isolate of AB, based on the seedling and adult plant reaction. QTL1 was mapped to LG3 linked to marker TR58 and explained 18.6% of the phenotypic variance (R ²) for AB resistance at the adult plant stage. QTL2 and QTL3 were both mapped to LG4 close to four SSR markers and accounted for 7.7% and 9.3%, respectively, of the total phenotypic variance for AB resistance at seedling stage. The SSR markers which flanked the AB QTLs were validated in a half-sib population derived from the same resistant parent ICCV 04516. Markers TA146 and TR20, linked to QTL2 were shown to be significantly associated with AB resistance at the seedling stage in this half-sib population. The markers linked to these QTLs can be utilized in marker-assisted breeding for AB resistance in chickpea.     

A SNP genetic linkage map based on the ‘Hamilton’ by ‘Spencer’ recombinant inbred line population identified QTL for seed isoflavone contents in soybean

Soybean is one of the most important crops worldwide for its protein and oil as well as the health beneficial phytoestrogens or isoflavone. This study reports a relatively dense single nucleotide polymorphism (SNP)‐based genetic map based on ‘Hamilton’ by ‘Spencer’ recombinant inbred line population and quantitative trait loci (QTL) for seed isoflavone contents. The genetic map is composed of 1502 SNP markers and covers about 1423.72 cM of the soybean genome. Two QTL for seed isoflavone contents have been identified in this population. One major QTL that controlled both daidzein (qDZ1) and total isoflavone contents (qTI1) was found on LG C2 (Chr 6). And a second QTL for glycitein content (qGT1) was identified on the LG G (Chr 18). These two QTL in addition to others identified in soybean could be used in soybean breeding to optimize isoflavone content. This newly assembled soybean linkage map is a useful tool to identify and map QTL for important agronomic traits and enhance the identification of the genes involved in these traits.     

QTL mapping for capsaicin and dihydrocapsaicin content in a population of Capsicum annuum ‘NB1’ × Capsicum chinense ‘Bhut Jolokia’

Capsaicinoids are pungent compounds used for industrial and medical purposes including food, medicine and cosmetics. The Indian local variety ‘Bhut Jolokia’ (Capsicum chinense Jacq.) is one of the world's hottest chilli peppers. It produces more than one million Scoville heat units (SHUs) in total capsaicinoids. In this study, our goal was to identify quantitative trait loci (QTLs) responsible for the high content of capsaicin and dihydrocapsaicin in ‘Bhut Jolokia’. Capsicum annuum ‘NB1’, a Korean pepper inbred line containing 14 000 SHUs, was used as a maternal line. An F₂ population derived by crossing between ‘NB1’ and ‘Bhut Jolokia’ was generated to map QTLs for capsaicinoids content. A total of 234 markers, including 201 HRM, 21 SSR, 2 CAPS and 10 gene‐based markers of the capsaicinoid synthesis pathway, were mapped. The final map covered a total distance of 1175.2 cM and contained 12 linkage groups corresponding to the basic chromosome number of chilli pepper. Capsaicin and dihydrocapsaicin content were analysed in 175 F₂ pepper fruits using the HPLC method. The maximum total capsaicinoids content was 1389 mg per 100g DW (dry weight), and the minimum content was 11 mg per 100g DW. Two QTLs (qcap3.1 and qcap6.1) for capsaicin content were identified on LG3 and LG6, and two QTLs (qhdc2.1 and qdhc2.2) for dihydrocapsaicin content were located on LG2. We did not detect QTLs for total capsaicinoids content. The QTL positions for capsaicin content were different from those for dihydrocapsaicin content. These results indicate that the complexity of selecting for more pungent chilli peppers must be considered in a chilli pepper breeding programme. The QTL‐linked markers identified here will be helpful to develop more pungent pepper varieties from ‘Bhut Jolokia’, a very hot pepper.     

Identification of QTL for oil content,seed yield,and flowering time in oilseed rape (<Emphasis Type="Italic">Brassica napus</Emphasis>)

A genetic map was constructed with 353 sequence-related amplified polymorphism and 34 simple sequence repeat markers in oilseed rape (Brassica napus L.). The map consists of 19 linkage groups and covers 1,868 cM of the rapeseed genome. A recombinant doubled haploid (DH) population consisting of 150 lines segregating for oil content and other agronomic traits was produced using standard microspore culture techniques. The DH lines were phenotyped for days to flowering, oil content in the seed, and seed yield at three locations for 3 years, generating nine environments. Data from each of the environments were analyzed separately to detect quantitative trait loci (QTL) for these three phenotypic traits. For oil content, 27 QTL were identified on 14 linkage groups; individual QTL for oil content explained 4.20–30.20% of the total phenotypic variance. For seed yield, 18 QTL on 11 linkage groups were identified, and the phenotypic variance for seed yield, as explained by a single locus, ranged from 4.61 to 24.44%. Twenty-two QTL were also detected for days to flowering, and individual loci explained 4.41–48.28% of the total phenotypic variance.     

Identification of QTLs for fruit quality traits in Japanese apples: QTLs for early ripening are tightly related to preharvest fruit drop

Many important apple (Malus × domestica Borkh.) fruit quality traits are regulated by multiple genes, and more information about quantitative trait loci (QTLs) for these traits is required for marker-assisted selection. In this study, we constructed genetic linkage maps of the Japanese apple cultivars ‘Orin’ and ‘Akane’ using F₁ seedlings derived from a cross between these cultivars. The ‘Orin’ map consisted of 251 loci covering 17 linkage groups (LGs; total length 1095.3 cM), and the ‘Akane’ map consisted of 291 loci covering 18 LGs (total length 1098.2 cM). We performed QTL analysis for 16 important traits, and found that four QTLs related to harvest time explained about 70% of genetic variation, and these will be useful for marker-assisted selection. The QTL for early harvest time in LG15 was located very close to the QTL for preharvest fruit drop. The QTL for skin color depth was located around the position of MYB1 in LG9, which suggested that alleles harbored by ‘Akane’ are regulating red color depth with different degrees of effect. We also analyzed soluble solids and sugar component contents, and found that a QTL for soluble solids content in LG16 could be explained by the amount of sorbitol and fructose.     

Development of high-density interspecific genetic maps for the identification of QTLs conferring resistance to <Emphasis Type="Italic">Valsa ceratosperma</Emphasis> in apple

Valsa canker (Valsa ceratosperma (Tode ex Fr.) Maire) is one of the most destructive fungal diseases of apple, especially in Eastern Asia. In this study, the first high density genetic linkage map of Malus asiatica × Malus domestica was constructed by 640 simple sequence repeats (SSRs) and 490 single nucleotide polymorphisms (SNPs), which spanned 1497.5 cM with an average marker interval of 1.33 cM per marker. Quantitative trait loci (QTLs) for apple’s resistance to V. ceratosperma isolates 03-8 and xc56 were identified using the linkage map. Lesion lengths were used as the phenotypic data for the QTL analysis, which were measured on 1-year-old shoots inoculated with conidia of the two isolates. One QTL for resistance to isolate 03-8 was mapped on LG 16, and one QTL for resistance to isolate xc56 was detected on LG 9. Our research not only promoted the further understanding of the genetic basis of apple’s resistance to Valsa canker but also provided two molecular markers that might be used in future marker-assisted selection for resistance in apple breeding programs.