QTL mapping and gene mining to identify genes on soybean (Glycine max) associated with NopB of Sinorhizobium fredii HH103

Soybean is a special crop that can utilize N₂ in the air via symbioses with Rhizobium spp. The formation of effective nodules is a complex process in which nodulation outer proteins (Nops) are determinants of establishment of a symbiotic relationship. We constructed a Sinorhizobium fredii HH103ΩnopB mutant. A nodulation test showed that the mutant had a negative effect on the Suinong14, ZYD00006, Dongnong594 and Charleston soybean lines. Recombinant inbred soybean lines were independently inoculated with the mutant and wild‐type strains, and five and four quantitative trait loci (QTLs) were identified by analysing the nodule number (NN) and nodule dry weight (NDW), respectively. We chose one QTL that overlapped with other studies and a novel QTL identified in our study and selected six candidate genes for further analysis. The qRT‐PCR analysis showed that only changes in Glyma.17G166200 expression depended on NopB. Further analysis showed that Glyma.17G166200 encoded a protein with a D‐glucose‐binding domain and a serine‐threonine/tyrosine protein kinase catalytic domain that was involved in the abscisic acid (ABA) pathway.     

Combining ability for nodulation in common bean (Phaseolus vulgaris L.) genotypes from Andean and Middle American gene pools

Ten F₁'s obtained from crosses among five common bean genotypes of Andean (WAF 15, Mineiro Precoce and Batatinha) and Middle American (BAT 304 and Ouro) gene pools were assessed for their combining abilities for root nodulation with Rhizobium tropici strain CIAT 899. The plants were grown under controlled conditions and evaluated for number of nodules per plant (NN), nodule dry weight (NDW), mean nodule weight (MNW) and plant fresh weight (PFW). The subdivision of the treatment effects on the general (GCA) and specific combining effects (SCA) were performed according to Griffing's diallel analysis method 2, model 1. The analyses of variance and estimates of quadratic components showed that non-additive gene effects were more important in the expression of NN and PFW, whereas additive gene effect was predominant for NDW and MNW. A close association was observed between high number of nodules and GCA. Generally, crosses involving parents of different gene pools yielded hybrids with high positive estimates of SCA for all assessed traits. The genotypes of Andean origin WAF 15 and Mineiro Precoce are the most promising parents for breeding programs to increase NN and NDW in common beans. This revised version was published online in July 2006 with corrections to the Cover Date.     

Identification of quantitative trait loci controlling soybean seed weight in recombinant inbred lines derived from PI 483463 (Glycine soja) × ‘Hutcheson’ (G. max)

The objective of this study was to identify quantitative trait loci (QTLs) controlling 100‐seed weight in soybean using 188 recombinant inbred lines (RIL) derived from a cross of PI 483463 and ‘Hutcheson’. The parents and RILs were grown for 4 years (2010–2013), and mature, dry seeds were used for 100‐seed weight measurement. The variance components of genotype (a), environment (e) and a × e interactions for seed weight were highly significant. The QTL analysis identified 14 QTLs explaining 3.83–12.23% of the total phenotypic variation. One of the QTLs, qSW17‐2, was found to be the stable QTL, being identified in all the environments with high phenotypic variation as compared to the other QTLs. Of the 14 QTLs, 10 QTLs showed colocalization with the seed weight QTLs identified in earlier reports, and four QTLs, qSW5‐1, qSW14‐1, qSW15‐1 and qSW15‐2, found to be the novel QTLs. A two‐dimensional genome scan revealed 11 pairs of epistatic QTLs across 11 chromosomes. The QTLs identified in this study may be useful in genetic improvement of soybean seed weight.     

Mapping quantitative trait loci (QTLs) underlying seed vitamin E content in soybean with main,epistatic and QTL × environment effects

Soybean (Glycine max (L.) Merr.) seed contains small amounts of tocopherol, a non‐enzymatic antioxidant known as lipid‐soluble vitamin E (VE). Dietary VE contributes to a decreased risk of chronic diseases in humans and has several beneficial effects on resistance to stress in plants, and increasing VE content is an important breeding goal for increasing the nutritional value of soybean. In this study, quantitative trait loci (QTLs) underlying VE content with main, epistatic and QTL × environment effects were identified in a population of F_5 : 6 recombinant inbred lines from a cross between ‘Hefeng 25’ (a low‐VE cultivar) and ‘OAC Bayfield’ (a high‐VE cultivar). A total of 18 QTLs were detected that showed additive main effects (a) and/or additive × environment interaction effects (ae) in different environments. Moreover, 19 epistatic pairs of QTLs were found to be associated with α‐tocopherol (α‐Toc), γ‐tocopherol (γ‐Toc), δ‐tocopherol (δ‐Toc) and total VE (TE) contents. The QTLs identified in multienvironments could provide more information about QTL by environment interactions and could be useful for the marker‐assistant selection of soybean cultivars with high seed VE contents.     

A single dominant gene in McCall soybean prevents effective nodulation with Rhizobium fredii USDA257

Summary Rhizobium fredii USDA257 will effectively nodulate Asiatic and unimproved soybean cultivars, such as Peking, but most of the highly selected North American cultivars, such as McCall, produce at most rudimentary, ineffective nodules. In R. fredii USDA 257, a locus containing 6 open reading frames is responsible for this cultivar specific incompatibility. To examine the genetic control of incompatability on the part of the host, the soybean cultivars Peking and McCall were crossed to produce five F₁ progeny. These plants and their selfed progeny were tested for nodulation with USDA257. Resistance to nodulation was found to be conditioned by a single dominant gene. These results indicate that, in soybean, strain specific resistance to nodulation can result from gene(s)-for-gene interactions.     

Dissection of component QTL expression in yield formation in rice

Yield and its components were investigated by using a population of 241 recombinant inbred lines (F₉ RILs) derived from an elite hybrid rice cross of ‘Zhenshan 97’בMinghui 63′. Quantitative trait loci (QTLs) for causal analysing of yield traits were detected at different yield component (YC) influences by conditional and unconditional QTL mapping methods. The number of QTLs significantly affecting yield was different at component‐special influence. Some QTLs controlling yield identified in one component influence were undetectable at the others. More QTLs for yield could be detected at different YC influences. It is possible to reveal that causal gene expression for yield could be different at different YC influences. Mapping QTLs for component effects of yield could help us in understanding the nature of cause‐effect traits for the formation of grain yield.     

Identification and validation of an over‐dominant QTL controlling soybean seed weight using populations derived from Glycine max × Glycine soja

Heterosis, or hybrid vigour, has been used to improve seed yield in several important crops for decades and it has potential applications in soybean. The discovery of over‐dominant quantitative trait loci (QTL) underlying yield‐related traits, such as seed weight, will facilitate hybrid soybean breeding via marker‐assisted selection. In this study, F₂ and F_2 : 3 populations derived from the crosses of ‘Jidou 12’ (Glycine max) × ‘ZYD2738’ (Glycine soja) and ‘Jidou 9’ (G. max) × ‘ZYD2738’ were used to identify over‐dominant QTL associated with seed weight. A total of seven QTL were identified. Among them, qSWT_13_1, mapped on chromosome 13 and linked with Satt114, showed an over‐dominant effect in two populations for two successive generations. This over‐dominant effect was further examined by six subpopulations derived from ‘Jidou12’ × ‘ZYD2738’. The seed weight for heterozygous individuals was 1.1‐ to 1.6‐fold higher than that of homozygous individuals among the six validation populations examined in different locations and years. Therefore, qSWT_13_1 may be a useful locus to improve the yield of hybrid soybean and to understand the molecular mechanism of heterosis in soybean.     

QTL mapping for seedling traits associated with low‐nitrogen tolerance using a set of advanced backcross introgression lines of rice

Nitrogen (N) deficiency is a major yield‐limiting factor in rice production. The objective of this study was to identify putative QTLs for low‐N stress tolerance of rice, using an advanced backcross population derived from crosses between an indica cultivar ‘93‐11’ and a japonica cultivar ‘Nipponbare’ and genotyped at 250 marker loci. Plant height, maximum root length, root dry weight, shoot dry weight and plant dry weight under two N conditions and their relative traits were used to evaluate low‐N tolerance at the seedling stage. A total of 44 QTLs were identified on chromosomes 1, 2, 3, 4, 5, 6, 8 and 9. Eight intervals on five chromosomes were identified to harbour multiple QTLs, suggesting pleiotropism or multigenic effects according to the contributor of alleles. Some QTL clusters were found in the nearby regions of genes associated with N recycling in rice, indicating that the key N metabolism genes might have effects on the expression of QTLs. Several unique QTLs for relative traits were detected, which suggested the specific genetic basis of relative performance.     

Fine mapping of a QTL for the number of spikelets per panicle by using near‐isogenic lines derived from an interspecific cross between Oryza sativa and Oryza minuta

We constructed a high‐resolution physical map for the qSPP7 QTL for spikelets per panicle (SPP) on rice chromosome 7 across a 28.6‐kb region containing four predicted genes. Using a series of BC₇F₄ near‐isogenic lines (NILs) derived from a cross between the Korean japonica cultivar ‘Hwaseongbyeo’ and Oryza minuta (IRGC Acc. No. 101144), three QTLs for the number of SPP, grains per panicle and primary branches were identified in the cluster (P ≤ 0.01). All three QTLs were additive, and alleles from the O. minuta parent were beneficial in the ‘Hwaseongbyeo’ background. qSPP7 was mapped to a 28.6‐kb region between the two simple sequence repeat (SSR) markers RM4952 and RM21605. The additive effect of the O. minuta allele at qSPP7 was 23 SPP, and 43.6% of the phenotypic variance was explained by the segregation of the SSR marker RM4952. Colocalization of the three QTLs suggested that this locus was associated with panicle structure and had pleiotropic effects. The NIL populations and molecular markers are useful for cloning qspp7.     

Quantitative trait loci with additive and epistatic effects underlying resistance to two HG types of soybean cyst nematode

The resistance of soybean (Glycine max L. Merr.) cultivars varies with the different races of the soybean cyst nematode (SCN), Heterodera glycines, referred to as HG types (biotypes). Resistant cultivars with durable resistance are emphasized in recent years. The aim here was to identify quantitative trait loci (QTLs) for resistance to two SCN HG types (HG type 2.5.7, race 1; and HG type 1.2.3.5.7, race 4) in resistant cultivar ‘L‐10’ and to analyse the additive and epistatic effects of the identified QTLs. A total of 140 F₅‐derived F₁₀ recombinant inbred lines (F_5:10 RILs) were advanced via single‐seed‐descent from the cross between ‘L‐10’ (broadly resistant to SCN) and “Heinong 37” (SCN‐susceptible). For SCN HG type 2.5.7 and HG type 1.2.3.5.7 resistance, three and six QTLs for resistance to SCN HG type 2.5.7 and HG type 1.2.3.5.7 were identified, respectively, most of which could explain <10% of the phenotypic variation. Among these QTLs, five were identified over 2 years, while the other QTLs were detected in either 2009 or 2010. QSCN1‐2, located near the SSR marker Sat_069 of linkage group D1b (Chromosome, 2), was responsible for the largest proportion of phenotypic variation (16.01% in 2009 and 18.94% in 2010), suggested that it could effectively be used as a candidate QTL for the marker‐assisted selection (MAS) of soybean lines resistant to SCN. Additionally, for SCN HG type 2.5.7 and HG type 1.2.3.5.7 resistance, two and four QTLs showed an additive effect (a), respectively. One epistatic pair of QTLs (QSCN1‐1‐QSCN1‐3) for SCN HG type 2.5.7 resistance and eight epistatic pairs of QTLs for SCN HG type 1.2.3.5.7 resistance were found to have significant aa effects, among which one pair of QTLs (QSCN4‐4 and QSCN4‐5) contributed a large proportion of aa effects (3%). The results indicated that additive and epistatic effects could significantly affect SCN resistance. Therefore, both of a and aa effects should be considered in MAS programmes.     

大豆株高QTL发育动态分析

应用分子遗传连锁图谱和条件QTL定位方法对性状进行动态分析是发育遗传学新的研究方法。采用来自Charleston ×东农594的143个重组自交系（RILs），构建了一个20条连锁群的大豆分子遗传连锁图谱，以此为基础，采用复合区间作图法共定位了28个显著影响株高发育的非条件QTL，以条件分析方法和复合区间作图法相结合定位了21个影响株高发育的条件QTL。不同发育时期显著影响株高的QTL数目和遗传效应的变化，说明控制株高发育的数量基因位点是选择性表达的。因此，进行标记辅助选择时综合考虑不同发育时期表达的QTL，才能取得较好的效果。     

Mapping of quantitative trait loci for resistance to race 1 of Pyrenophora tritici‐repentis in synthetic hexaploid wheat

Tan spot, caused by a necrotrophic fungus Pyrenophora tritici‐repentis (Ptr), has become an important foliar disease of wheat worldwide. Effective control of tan spot can be achieved by deployment of resistant wheat cultivars. An F_2:3 population derived from a cross between synthetic hexaploid wheat (SHW), TA4161‐L1 (moderately resistant) and susceptible winter wheat cultivar, ‘TAM105’ was evaluated with race 1 of Ptr under controlled conditions. The population was genotyped using Diversity Arrays Technology (DArT). Presence of transgressive segregants indicated contribution of positive alleles from both parents. Two major QTLs were located on the short arm of chromosomes 1A and 6A and designated as QTs.ksu‐1A and QTs.ksu‐6A, respectively. Two additional QTLs were identified on chromosome 7A. Resistant alleles of all the QTLs were contributed by TA4161‐L1. Novel QTLs on 6A and 7A can be a valuable addition to known resistance genes and utilized in breeding programmes to produce highly resistant cultivars.     

The Tripartite Symbiosis Formed by Indigenous Arbuscular Mycorrhizal Fungi, Bradyrhizobium japonicum and Soya Bean Under Field Conditions

The tripartite symbiosis formed by indigenous arbuscular mycorrhizal fungi (AMF), Bradyrhizobium japonicum (Kirchner) Jordan and soya bean (Glycine max L. Merr. cv. Evans) was investigated under field conditions to test the hypotheses that: (i) the tripartite symbiosis enhances nodulation and nodule activity; and (ii) its establishment does not rely on improved phosphorus (P) uptake through the fungal partner. Soil tillage was used to produce treatments with contrasting AMF colonization potentials while the amount of B. japonicum inoculum was kept constant. Nodulation, AMF colonization and the P and nitrogen (N) nutrition of plants were evaluated at 10 and 51 (full‐bloom) days after emergence. N₂ fixation was estimated by the difference method and by the isotopic dilution method. At the early stage of plant growth, AMF hyphal colonization and nodulation were, respectively, 16 % and 33 % greater in plants from untilled than from rototilled soil. The establishment of the tripartite symbiosis was observed under field conditions, and factors other than P nutrition were critical to its formation. However, the tripartite symbiosis did not promote N₂ fixation under the high soil P conditions of this study.     

Identification of QTLs for red fruit firmness using the wild tomato species Solanum pennellii LA716 introgression lines

The introgression lines (ILs) of the wild tomato species Solanum pennellii have been widely used to identify genes related to yield, texture, disease resistance and stress tolerance. In addition to flavour, fruit firmness is an important evaluation index and essential trait indicating tomato fruit quality. Quantitative trait loci (QTL) for fruit firmness have been identified through hand squeezing and pericarp puncturing. However, these techniques hardly reveal the force value of the whole fruit suffering from rupture or deformation. In this study, S. pennellii ILs were used to identify QTLs related to fruit firmness through flat‐plate compression. Nine QTLs for enhancing and sixteen QTLs for decreasing fruit firmness were successfully identified. Compared with that of ‘M82’, the amount of QTLs that enhance fruit firmness increased by 8.76% to 21.00%, and the amount of QTLs that reduce fruit firmness decreased by ?8.27% to ?30.80%. The QTL Crf12a and Crf‐R‐7b is the strongest and weakest QTL, respectively, while they are very stable in all independent biological trials. Six QTLs should be further confirmed through open‐field trials.     

Heterosis of interspecific soybean hybrids for traits related to N2 fixation

Summary Three greenhouse experiments were conducted to compare the performance of soybean (Glycine max (L.) Merr.), wild soybean (G. soja Sieb. et Zucc.), and soybean x wild soybean hybrids for traits relating to N₂ fixation including nodulation, acetylene reduction, nodule leghemoglobin concentration, and nitrogen (N) accumulation and dry matter (DM) accumulation. In all three experiments G. max generally exceeded G. soja in nodulation, acetylene reduction, and N and DM accumulation while the soybean possessed higher nodule leghemoglobin concentration. In Experiment I, the mean of the hybrids did not differ significantly from the G. max parent in nodule mass, acetylene reduction activity, nodule leghemoglobin concentration, or DM accumulation. The hybrids did exceed the soybean parent in N accumulation, thus demonstrating high parent heterosis. In Experiments IIA and IIB with a more carefully chosen set of G. soja parents, high parent heterosis of individual crosses was common. Across the three experiments average high parent heterosis was 34, 28, and 28%, respectively, for nodule mass, N accumulation, and DM accumulation. If one accepts the assumption that hybrid vigor results from the accumulation of dominant alleles, then these alleles could theoretically be accumulated via selection in a homozygous genotype. If this is true than the results of the experiments reported here suggest that interspecific soybean x wild soybean crosses could serve as sources of homozygous lines which would exceed currently available soybean cultivars in nodule mass, and N and DM accumulation.This work was supported in part by the USDA Competitive Grants Program, Grant 82-CRCR-1-1039 and Cooperative Agreement 58-32U4-2-370 between the U.S. Department of Agriculture and the Agronomy Department, University of Maryland, Scientific Article No. A-4648, Contribution No. 7644 of the Maryland Agricultural Experiment Station.     

Deployment of cold tolerance loci from Oryza sativa ssp. Japonica cv. ‘Nipponbare’ in a high‐yielding Indica rice cultivar ‘93‐11’

Cold tolerance is a complex trait, and QTL pyramiding is required for rice breeding. In this study, a total of seven QTLs for cold tolerance in the Japonica rice variety ‘Nipponbare’ were identified in an F_2:3 population. A stably inherited major QTL, called qCTS11, was detected in the region adjacent to the centromere of chromosome 11. In a near‐isogenic line population, the QTL was further dissected into two linked loci, qCTS11.1 and qCTS11.2. Both of the homozygous alleles of qCTS11.1 and qCTS11.2 from ‘Nipponbare’ showed major positive effects on cold tolerance. Through pyramiding the linked QTLs in the cold‐sensitive Indica rice cultivar ‘93‐11’, we have developed a new elite, high‐yielding Indica variety with cold tolerance.     

Quantitative trait loci mapping of adult‐plant resistance to leaf rust in a Fundulea 900 × ‘Thatcher’ wheat cross

Wheat leaf rust (LR), caused by the obligate biotrophic fungus Puccinia triticina (Pt), is a destructive foliar disease of common wheat (Triticum aestivum L.) worldwide. The most effective, economic means to control the disease is resistant cultivars. The Romanian wheat line Fundulea 900 showed high resistance to LR in the field. To identify the basis of resistance to LR in Fundulea 900, a population of 188 F_2:3 lines from the cross Fundulea 900/‘Thatcher’ was phenotyped for LR severity during the 2010–2011, 2011–2012 and 2012–2013 cropping seasons in the field at Baoding, Hebei Province. Bulked segregant analysis and simple sequence repeat markers were used to identify the quantitative trait loci (QTLs) for LR adult‐plant resistance in the population. Three QTLs were detected and designated as QLr.hebau‐1BL, QLr.hebau‐2DS and QLr.hebau‐7DS. Based on the chromosome positions and molecular marker tests, QLr.hebau‐1BL is Lr46, and QLr.hebau‐7DS is Lr34. QLr.hebau‐2DS was derived from ‘Thatcher’ and was close to Lr22. This result suggests that Lr22b may confer residual resistance on field nurseries when challenged with isolates virulent on Lr22b, or another gene linked to Lr22b confers this resistance from ‘Thatcher’. This study confirms the value of Lr34 and Lr46 in breeding for LR resistance in China; the contribution of the QTL to chromosome 2D needs further validation.     

Breeding and characterization of soybean Triple Null; a stack of recessive alleles of Kunitz Trypsin Inhibitor,Soybean Agglutinin,and P34 allergen nulls

Soybean (Glycine max) seeds contain bioactive proteins with antinutritional and immunological properties that affect metabolism and assimilation of nutrients. The presence of antinutritional proteins requires soybeans to be heat‐treated resulting in input energy costs. Nulls for bioactive seed proteins have been previously isolated from the USDA soybean collection, including Kunitz trypsin inhibitor (TI), soybean agglutinin (LE) and immunodominant soybean allergen P34 protein. Each of these nulls has the potential to partially address the concerns of soybean feed/food consumption. A stack of recessive nulls of TI, LE and P34 was created in a cv ‘Williams 82’ background termed ‘Triple Null’. Triple Null has a slight reduction of total protein compared with ‘Williams 82’ corresponding to aggregate contribution of TI, LE and P34 in the seed proteome. Triple Null's proteome analysis revealed P34 and TI nulls are frame‐shift mutants able to accumulate small amounts of authentic P34 and TI proteins. Triple Null has possible application as a conventional feed/food source and for immunotherapy to mitigate soybean allergenic response.     

The transition time from the juvenile phase to the adult phase differs in soybean cultivars ‘Enrei’ and ‘Peking’

Plants develop juvenile phase to adult phase in vegetative stage. Although soybean is a very important crop worldwide, there has been only one study of the juvenile–adult phase change. In this study, we determined that the juvenile–adult phase change occurred at different stages in two soybean cultivars that differ in their photosensitivity. Cultivar ‘Enrei’ (E1e2e3E4) is weakly photosensitive and cultivar ‘Peking’ (E1E2E3E4) is strongly photosensitive. In ‘Enrei’, the leaf size gradually increased at a constant leaf position regardless of the difference in day length. In ‘Peking’ plants transferred to short‐day conditions at several leaf development stages, leaf size gradually increased at different leaf positions. Expression of miR156 by ‘Enrei’ transferred to short‐day conditions had nearly the same pattern as that of ‘Enrei’ grown under long‐day conditions. In ‘Peking’, the expression of miR156 had different patterns in younger leaves of plants subjected to either a short‐day treatment or long‐day conditions. These results indicate that the E2 and E3 loci that regulate photosensitivity also regulate the expression of miR156 and the juvenile–adult phase change in soybean.