Inheritance and gene tagging of male fertility restoration of cytoplasmic-nuclear male-sterile line NJCMS1A in soybean

The F₁, F₂ and F_2:3 of the NJCMS1A × 'Zhongdou 5' cross were used to analyse the inheritance of the male fertility restoration of the cytoplasmic-nuclear male-sterile line NJCMS1A in soybean. The results of genetic analysis showed two pairs of dominant genes conferring the male fertility restoration of NJCMS1A, which further confirmed previous results. The F₂ population from the NJCMS1A × 'Zhongdou 5' cross was used for tagging the restorer genes for NJCMS1A with 664 pairs of simple sequence repeat primers selected randomly from the genetic linkage map of soybean published by Cregan et al. (1999) . Satt626 on linkage group M and Satt300 on linkage group A1 of the integrated linkage map by Song et al. (2004) were found to link to the two restorer genes of NJCMS1A. The maximum-likelihood estimates of the genetic distance between the two markers, Satt626 and Satt300, and the two restorer genes of 'Zhongdou 5' were 9.75 and 11.18 cM, respectively.     

QTL analysis on plant height and flowering time in Brassica napus

To identify quantitative trait loci (QTL) controlling plant height (PH) and flowering time (FT), a high stalk/later bloom Brassica napus line '2091' was crossed with a dwarf/early bloom mutant '99CDAM'. A segregating population with 145 F_2:3 lines was constructed and grown in field for 2 years. Based on this population, a linkage map consisting of 199 amplified fragment length polymorphism and 42 simple sequence repeat loci was constructed. The LOD threshold of 3.0 was used for declaring the existence of putative QTL. In total, seven QTL related to PH were identified, accounting for 8.5–28.6% of phenotypic variation, respectively, of which two QTL were identified in both years. Six QTL for FT were detected, accounting for 8.1–30.4% of phenotypic variation individually, of which two QTL were identified in both years. The main QTL of PH and FT were both mapped on linkage group 13, with overlapped creditable regions. QTL ph17 and ft17 existed at the same location. These might be the genetic base for the significant correlation observed between PH and FT.     

玉米雄穗分枝数主效QTL定位及qTBN5近等基因系构建

立足于发掘玉米雄穗分枝数优异基因资源, 利用郑单958骨干亲本郑58和昌7-2构建的188个重组自交系(recombinant inbred line, RIL)家系群体, 结合288个多态性分子标记构建的连锁图谱和2年玉米雄穗分枝数表型数据, 运用完备复合区间作图法进行QTL定位, 共检测到5个控制玉米雄穗分枝数的一致性主效QTL, 分别位于玉米5条染色体上。通过连续回交及分子标记辅助选择构建了位于bin 5.05的控制雄穗分枝数主效QTL-qTBN5近等基因系(near isogenic line, NIL), 对基因遗传效应进行了验证, 并将qTBN5进一步定位在13.2 Mb区间之内, 为玉米雄穗分枝数主效基因的精细定位及分子育种奠定基础。     

Genetic linkage map of cassava (Manihot esculenta Crantz) based on AFLP and SSR markers

To generate a genetic linkage map of cassava ( Manihot esculenta Crantz), 58 F₁ progenies from a cross between Rayong 90 (female) and Rayong 5 (male) were examined in amplification fragment length polymorphism (AFLP) and simple sequence repeat (SSR) analyses. A total of 469 polymorphic markers consisting of 378 AFLPs generated from 76 primer combinations and 91 SSRs were identified. These markers were analyzed using the joinmap ^® 3.0 package program to construct a genetic linkage map. A total of 33 linkage groups of a common map were constructed from 119 AFLPs and 18 SSRs, spanning 1095 cM with an average of 7.99 cM between markers. The genetic linkage map generated in this study will be useful for genetic studies in cassava particularly for the identification of genetic markers linked to traits of interest, although the complex cassava genome suggests that maybe a long term objective.     

Identification and molecular tagging of a stripe rust resistance gene in wheat line P81

Stripe rust, caused by Puccinia striiformis f. sp. tritici (PST), is one of the most devastating diseases in common wheat ( Triticum aestivum L.). With the objective of identifying and tagging a new gene for resistance to stripe rust in wheat line P81, F₁, F₂ and F_2:3 populations from the cross 'Chuanmai 28'/P81 were inoculated with Chinese PST race CYR32 in greenhouse and field trials. P81 carried a single dominant gene for resistance (designated YrP81 ) to CYR32. Tests of allelism showed that YrP81 was different from Yr5 , Yr10 , Yr15 and Yr26 . Simple sequence repeat (SSR) and resistance gene-analogue polymorphism (RGAP) between the parents were used for genotyping the F₂ populations. YrP81 was closely linked to four SSR loci on chromosome 2BS with genetic distances of 18.3 cM ( Xwmc25 ), 1.8 cM ( Xgwm429 ), 4.1 cM ( Xwmc770 ) and 5.3 cM ( Xgwm148 ). Two RGAP markers RGA1 (NLRR/XLRR) and RGA2 (Pto kin4/NLRR-INV2) were also closely linked to YrP81 with genetic distances of 4.7 and 6.3 cM, respectively. The linkage map of YrP81 and molecular markers was established in the order Xwmc25 - RGA2 - RGA1 - Xgwm429 - YrP81 - Xwmc770 - Xgwm148 . Pedigree analysis, response patterns with Chinese PST races and associations with markers suggested that YrP81 is a novel stripe rust resistance gene. The PCR-based microsatellite and RGAP markers identified here could be applied in selection of YrP81 in wheat breeding.     

Co-localization of quantitative trait loci for foliar disease resistance in sorghum

Quantitative trait loci (QTL) analysis of resistance to three foliar diseases, viz. target leaf spot, zonate leaf spot and drechstera leaf blight was undertaken in sorghum using 168 F₇ recombinant inbred lines derived from a cross between '296B' (resistant) and 'IS18551' (susceptible) parents. The genomic region flanked by plant colour locus ( Plcor ) and simple sequence repeat marker Xtxp95 on chromosome SBI-06 harboured disease-response QTL for all the three diseases caused by different fungal pathogens. It is hypothesized that this region on sorghum chromosome SBI-06 could harbour a cluster of disease-response loci to different pathogens as observed in the syntenic regions on rice chromosome 4 and maize chromosome 2. The information gained in this study can be used in deploying marker-assisted selection for foliar resistance and map-based isolation of important disease resistance genes in sorghum.     

Quantitative trait loci for stay-greenness and agronomic traits provide new insights into chlorophyll homeostasis and nitrogen use in rice

Functional stay-green (FSG) is characterized by delayed senescence during the reproductive stage in rice. A recombinant inbred line (RIL) population derived from ‘Pusa 677’/‘PSG16’ was used to study the genetics of FSG in a rice mutant, ‘PSG16’. The RILs exhibited significant variation under two N regimes for agro-morphological traits, leaf chlorophyll content, flowering time, yield components and nitrogen (N) use. Using a genome wide linkage map spanning 1910.8 cM with 104 polymorphic markers, we have mapped six major quantitative trait loci (QTLs). One robust QTL on chromosome 1 was associated consistently across different N regimes with chlorophyll content and flowering time. The QTL on chromosome 7 was associated with grain number, whereas the QTL on chromosome 6 was found related to N harvest index and spikelet fertility. Although ‘PSG16’ showed a clear advantage in grain yield as well as having better N assimilation, we could not establish a direct genetic relationship SG trait and N use. Based on the QTL information, FSG trait of ‘PSG16’ could be useful in deciphering multiple stress responses in rice.     

Identification of quantitative trait loci for drought tolerance at seedling stage by screening a large number of introgression lines in maize

The maize genome hosts tremendous phenotypic and molecular diversity. Introgression lines (ILs), developed by continuous backcrossing to recurrent parents, could provide a unique genetic stock for quantitative trait locus (QTL) mapping. Using maize lines from six heterotic groups of different ecological zones, we developed >500 BC₂F₂ IL sets by crossing 11 inbred lines (as recurrent parents) with >200 local maize inbred lines (as donor parents). Of them, 34 IL sets were selected as a subset for drought tolerance screening and a total of 417 ILs survived under severe water stress at seedling stage. One set of 32 surviving ILs, derived from Chang7-2/DHuang212, was used for QTL mapping with simple sequence repeat markers covering the whole genome, with seven QTL detected. Furthermore, investigating all surviving ILs, we identified two common regions in bin 3.04, corresponding to marker intervals bnlg1904–umc1772 and umc1223–bnlg1957, respectively, which shared high genetic variation in three IL sets. Our results indicated that selective genotyping can be used to identify genetic loci for complex traits. The ILs, highly selected for drought tolerance in this study, provide a unique set of materials for both genomic studies and development of enhanced germplasm resources.     

陆地棉光合相关性状的QTL定位分析

光合作用是棉花产量的主要物质来源。本研究以高光效陆地棉冀优861和低光效陆地棉新陆早25号为亲本组配的196个F₂单株为作图群体,利用SSR（Simple Sequence Repeat）标记构建陆陆杂交遗传连锁图谱,共有30个标记位点连锁,包含4个连锁群,全长244.4cM。利用QTL IciMapping 4.1软件的完备区间作图法对冀优861×新陆早25号F_2:3家系的光合相关性状进行QTL作图分析,共定位到光合相关5个性状的10个QTLs,其中1个光合速率QTL和1个胞间CO₂浓度QTL分别定位在D3和D7染色体上。本研究为棉花光合相关性状QTL的精细定位及分离克隆打下基础,为聚合棉花高光效分子标记辅助育种提供理论依据。     

Development of STS markers and QTL validation for common bacterial blight resistance in common bean

Common bacterial blight (CBB) of common bean ( Phaseolus vulgaris L.), is one of the major diseases that decrease yield and quality. A major quantitative trait locus (QTL) for CBB resistance from line XAN 159 was transferred into two bean lines, HR45 and HR67. Previous studies identified that two markers are linked to this QTL but the chromosome location was not consistent. To identify more tightly linked markers and to verify the chromosome location, 65 additional markers were mapped using 81 recombinant inbred lines (RILs) derived from a cross HR67 × OAC95-4. The QTL was mapped to a 13 cM region on chromosome 1 and defined by eight molecular markers that explained 25–52% of the phenotypic variation. Six tightly linked amplified fragment length polymorphism markers (0.6–9.7 cM from the QTL peak) were converted into seven sequence tagged site markers, three of which were mapped to this QTL. Five tightly linked markers were used to screen 907 F₂ plants derived from a cross HR45 × 'OAC Rex' and four of them were linked to each other within 4.2 cM. These markers may be useful in marker-assisted selection and map-based cloning of this major QTL.     

Quantitative trait loci analysis for resistance against Turnip mosaic virus based on a doubled-haploid population in Chinese cabbage

Turnip mosaic virus (TuMV) as the major virus infecting Brassica crops, often cause severe yield and quality losses in Chinese cabbage production in China. In this study, quantitative trait loci (QTL) analysis for TuMV resistance was conducted with a population of 100 doubled-haploid lines derived from the F₁ between 91-112 (resistant) and T12-19 (susceptible) through microspore culture. A total of 376 molecular markers including 235 amplified fragment length polymorphisms, 129 random amplified polymorphic DNAs, 10 simple sequence repeats, one SCAR and one morphological marker were employed to construct a linkage map with 10 linkage groups covering 809.1 cM with an average distance of 2.2 cM between loci. Resistance was assessed by artificial inoculation at the seedling stage and at the adult stage in field conditions, respectively. Four QTLs controlling TuMV resistance were identified with JoinMap QTL 4.0 and interval mapping method. Two QTLs ( Tu1 , Tu2 ) were associated with resistance at the seedling stage, each accounting for 58.2% and 14.7% of the phenotypic variation, respectively. Two QTLs ( Tu3 , Tu4 ) were found corresponding to the disease resistance at the adult plant stage, explaining 48.5% and 32.0% of the phenotypic variation, respectively. Marker-assisted selection for these major QTLs involved in TuMV resistance could be useful in Chinese-cabbage breeding programmes.     

基于两个陆地棉低世代群体定位纤维品质相关QTL

【目的】定位棉花纤维品质性状相关的数量性状位点(Quantitative trait locus,QTL)。【方法】以陆地棉高强纤维品系中棉所679和纤维品质一般的农垦5号为亲本构建包含200个单株的F2群体及对应的F2:3家系群体,对2个群体的纤维长度、断裂比强度等5个纤维品质性状进行检测。用6 688对简单重复序列(Simple sequence repeat, SSR)引物在双亲间筛选,得到149对多态性引物,以F2为作图群体,使用QTL IciMapping软件进行连锁图谱构建,并对F2及F2:3群体进行QTL定位。【结果】根据F2群体基因型信息构建了1张包含119个标记、28个连锁群、总长为1 173.5 cM(centiMorgan)的遗传连锁图谱。分别在F_2、F2:3群体中检测到9个和11个与纤维品质性状相关的QTLs,这些QTLs分布在11个连锁群上。其中F2群体的qFL-D11-1、q BT-D11-1与F2:3群体的qFL-D11-1、q MIC-D11-1均定位在标记DPL0062与HAU0423之间,推测这些位点可能是控制纤维品质性状的重要QTL。【结论】利用多个群体进行QTL定位有益于发现稳定的QTL位点,控制纤维品质性状的基因可能成簇存在,为挖掘纤维品质性状相关基因及分子标记辅助育种奠定基础。     

玉米产量相关性状QTL通用图谱的构建及功能候选基因的开发

通过生物信息学手段,收集整理玉米基因组数据库中产量相关性状的QTL信息,借助高密度玉米遗传连锁图谱IBM 2005 Neighbors和临近分子标记,建立了涵盖干物质和淀粉产量、籽粒相关性状、粒重相关性状以及穗部相关性状四大类共计18个指标的QTL通用图谱。并将位于QTL分布密集区域的基因,与水稻基因序列进行比对,找到了与水稻木质素合成有关的gh2具有高同源性的基因AY109876,同时改进了利用生物信息学手段筛选功能候选基因的路线图。     

Molecular mapping of a fertility restorer gene for cytoplasmic male sterility in soybean

In this study, we report the mapping of the Rf locus in soybean by microsatellite simple sequence repeat (SSR) genetic markers. A cross was made between cytoplasmic male sterility (CMS) line JLCMS82A and restorer line JIHUI 1 based on the DNA polymorphisms revealed by 109 SSR markers. A F₂ population derived from a single F₁ plant containing 103 individuals was used for mapping the Rf locus. The Rf gene of JIHUI 1 gametophytically restores male fertility to JLCMS82A. Fertile and semi-fertile DNA bulks and parental DNAs were screened with 219 SSR markers, and Satt215 which was previously mapped to soybean LG J, was found linked to the Rf gene. Five additional polymorphic SSR markers from LG J were used for analysis and a regional linkage map around the Rf locus was established. SSR markers, Sctt011 and Satt547, flanked the Rf locus at 3.6 cM and 5.4 cM, respectively. The availability of these SSR markers will facilitate the selection of restorer lines in hybrid soybean breeding.     

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.     

Relationships between seed oil content and fatty acid composition in high stearic acid sunflower

The high stearic acid sunflower mutant CAS-3 is characterized by a low seed oil content, which might represent a constraint for the commercial production of high stearic acid sunflower oil. The objective of the present research was to investigate the relationships between fatty acid profile and seed oil content in CAS-3. Plants of CAS-3 were reciprocally crossed with plants of breeding line ADV-37, with high oil content and standard fatty acid profile. Oil content and fatty acid composition were measured in individual F₂ seeds and F₂ plants (F₃ seeds averaged). Both F₂ seeds and F₂ plants from the cross ADV-37 × CAS-3 had a significantly higher oil content than those from the reciprocal cross, which indicated the existence of cytoplasmic effects in the genetic control of the trait. A consistent negative correlation between oil content and palmitic acid and a positive correlation between oil content and oleic acid were detected both in F₂ seeds and F₂ plants. Conversely, no consistent correlation between oil content and stearic acid was observed, which suggested the feasibility of simultaneous selection for both traits.     

Identification of a SCAR marker linked to a recessive male sterile gene (Tems) and its application in breeding of marigold (Tagetes erecta)

In marigold, an F₂ segregation population of 167 plants was constructed from a cross of a line (M525A) carrying the male sterility trait × an inbred line (f53f). In line M525A, the male sterility trait was controlled by the recessive gene, Tems . The intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) techniques combined with bulked segregant analysis were used to develop markers linked to the trait. From a survey of the 38 ISSR primers and 170 SRAP primer combinations, only one SRAP marker that was closely linked to the target trait was identified and successfully converted into sequence characterized amplified region (SCAR) marker that was located within 2.4 cM from Tems locus. The marker was validated with five other two-type lines and in each case the male fertile plants were reliably identified. This SCAR marker therefore permits the efficient marker-assisted selection of male sterile individuals in breeding programmes of marigold and will greatly facilitate the breeding of F₁ cultivars.     

Mapping of QTL for resistance to powdery mildew and resistance gene analogues in perennial ryegrass

The objective of this study was to map resistance gene analogues (RGA) and quantitative trait loci (QTL) for powdery mildew resistance in perennial ryegrass ( Lolium perenne  L.). The mapping population consisted of 184 F₂ genotypes produced from a cross between one genotype of a synthetic perennial ryegrass variety Veyo and one genotype from the perennial ryegrass ecotype Falster. The greenhouse infection was measured as number of sporulating colonies on a 10-cm leaf area with the maximum possible number of colonies to be counted set to 50. The range of infection scores in the population was 0–50 and the heritability was 71.7%. In total, two QTL for powdery mildew resistance were identified, and mapped to linkage groups (LG) LG3 and LG7. Individually, these QTL explained between 7.5% and 22.1% of the total phenotypic variation. Six RGA and three laccases were mapped to LG2, LG3, LG4, LG5 and LG7, however, no close linkage between RGA and QTL was observed.     

Mapping and validation of the quantitative trait loci for leaf stay‐green‐associated parameters in maize

Functional stay‐green is generally regarded as a desirable trait of varieties in major crops including maize. In this study, we used an F_3:4 recombinant inbred line population with 165 lines from a cross between a stay‐green inbred line (Zheng58) and a model inbred line (B73) using 211 polymorphic simple sequence repeat markers to map quantitative trait loci for three stay‐green‐associated parameters, chlorophyll content, photosystem II photochemical efficiency and stay‐green area, at maturity stage, detected a total of 23 quantitative trait loci (QTL) on nine chromosomes. Single QTL explained 3.7–13.5% of the phenotypic variance. Additionally, we validated some important stay‐green QTL using a heterogeneous inbred family approach and found that the stay‐green‐associated parameters were significantly correlated with the plant yield. This study may contribute to a better insight into the regulatory mechanism behind leaf stay‐green in maize and a novel development of elite maize varieties with delayed leaf senescence through molecular marker‐assisted selection.     

玉米叶片紫色中脉性状的遗传

利用引进的紫黑色种皮玉米随机交配群体,通过杂交和多代单株自交选择,培育出紫色中脉稳定纯系06Z-2。以06Z-2(P₂)和白色中脉自交系L-24(P₁)为材料,创制出F₁、F₂、BC₁和BC₂群体,对玉米叶片紫色中脉性状的遗传特征进行了研究。结果表明,紫色中脉性状属于显性单基因遗传,分离世代中紫色中脉株数与白色中脉株数的分离比符合孟德尔经典遗传学第一定律,且遗传稳定,易于转育。深入开展玉米叶片紫色中脉性状的遗传与应用研究,对于玉米自交系或杂交种防杂保纯、进行种子纯度鉴定、保护知识产权等具有重要的理论与适用价值。