Identifying AFLP and microsatellite markers for vernalization response gene Vrn-B1 in hexaploid wheat using reciprocal mapping populations

Marker‐assisted selection may be useful for combining specific vernalization response (Vrn) alleles into a single wheat genotype for yield enhancement; however, DNA markers are only available for two of the three genes identified to date. The objectives of this study were to investigate reciprocal effects on days to heading using F₂ populations generated by cross‐hybridizing near‐isogenic lines (NILs) carrying spring (Vrn‐B1; TDB) and winter (vrn‐B1; TDC) alleles, and to identify markers linked to Vrn‐B1 through genetic linkage analysis. Heading data were recorded for 91 and 89 progeny from reciprocal mapping populations TDB/TDC and TDC/TDB, respectively, and significant (P < 0.0001) reciprocal and dominance effects were detected. Among 207 amplified fragment length polymorphisms primer pairs and seven wheat microsatellite markers screened, two and one, respectively, were linked distally to Vrn‐B1 on wheat chromosome 5BL. Microsatellite Xgwm408 was most closely linked to Vrn‐B1 at 3.9 and 1.1 cM in the TDB/TDC and TDC/TDB map, respectively. Reciprocal differences in recombination distances emphasize the importance of female parent choice when generating mapping populations. Molecular markers are now available for three Vrn loci in wheat.     

Genetic structure of putative heterotic populations of alfalfa

Semi‐hybrids between genetically distant alfalfa (Medicago sativa subsp. sativa) populations may display heterosis whose extent is affected by the structure of genetic diversity across populations. This study aimed to assess the genetic diversity across three putative heterotic populations, one Italian, one Egyptian and one of semi‐erect germplasm from Eastern Europe, Canada and Spanish Mielga (EECM population). Each population was bred from ten parents after various selection cycles. Fifteen genotypes per population were characterized by 20 polymorphic SSR markers. The among‐population variance was over eightfold smaller than the average within‐population variance (2.05 vs. 17.24) and accounted for 10.6% of the total variation. G’_ST = .090 across markers indicated modest population differentiation. Various diversity measures, multidimensional scaling, and cluster analysis of the genetic structure indicated that the Italian population was more distant from the EECM population than the Egyptian one. The EECM and Egyptian populations were the most distant geographically and genetically. EECM displayed widest intrapopulation variation, accordingly to its constitutive geographical diversity. In conclusion, this study indicates modest genetic differentiation between alfalfa populations even for geographically distant germplasm.     

Genetic diversity evolution through participatory maize breeding in Portugal

Natural, and in particular, artificial (human) selection may pose a danger to the existing crop genetic diversity. Nevertheless, on-farm breeding systems seem to achieve phenotypic improvements even though preserving variability. Using SSR markers, we analysed several selection cycles, over a 20 years period, of a Portuguese on-farm participatory maize OPV-‘Pigarro’ breeding project. No significant differences in allelic richness (N_ar), observed heterozygosity (H_O), expected heterozygosity (or gene diversity; H_E) or inbreeding coefficient (f) were detected among the selection cycles. 58 out of 107 alleles were common to all the selection cycles studied. The analysis of molecular variance showed that the variation among selection cycles represented only 7% of the total molecular variation. However, the number of private alleles varied among the selection cycles, being the highest detected at the beginning of the selection project. These findings demonstrate that an allele flow took place during the on-farm selection process of ‘Pigarro’ but the level of genetic diversity was not significantly influenced. Since interesting phenotypic improvements were also achieved, on-farm breeding projects, like this one, should be valued as a way to preserve unique Portuguese maize landraces in risk of disappearing.     

Experimental evaluation of several cycles of marker-assisted selection in maize

A program was initiated in 1994 to compare the efficiency of marker-assisted selection (MAS) and conventional phenotypic selection. A population of 300 F_3:4 families was generated from the cross between two maize inbred lines F2 and F252 and selected on an index combining grain yield and grain moisture at harvest. This population was characterised for 93 RFLP markers and evaluated as testcrosses in a large range of environments. Three methods of selection were applied (i) two cycles of conventional phenotypic selection; (ii) two cycles of MAS based on an index combining phenotypic values and QTL genetic values and (iii) one cycle of combined MAS followed by two cycles of selection based only on the QTL effects estimated in the first generation. The different populations were characterised for RFLP markers. The evolution of allele frequencies showed that selection on only-markers was very efficient for fixing QTL alleles found favourable in the initial population. This evolution was quite different from that observed for phenotypic selection or combined MAS. Genetic gain was evaluated and found significant for each method of selection. Nevertheless, the difference between phenotypic selection and combined MAS was not significant. The two additional cycles of MAS on only-markers did not improve significantly the genetic value of the population. Moreover, the genetic variance of this population remained high, despite most of the QTL initially detected were almost fixed. The results suggest that the QTL effects estimated in the initial population were not stable due to epistasis and/or QTL by environment interactions. This revised version was published online in July 2006 with corrections to the Cover Date.     

Recurrent selection of cocoa populations in Côte d'Ivoire: comparative genetic diversity between the first and second cycles

In Côte d'Ivoire, the cocoa breeding programme has been based on the creation of hybrids between different genetic groups. From 1990 onward, a reciprocal recurrent selection programme has been set up with the purpose of improving simultaneously the characteristics of the two main genetic groups: Upper Amazon Forastero (UA) and a mixture of Lower Amazon Forastero (LA) and Trinitario (T). Based on data obtained from 12 microsatellite primers, the genetic diversity and genetic distances of the parental populations used in the first and second selection cycles are presented. The results revealed that the diversity of populations UA0 and UA1 on the one hand and (LA+T)0 and (LA+T)1 on the other is similar. The genetic distances were small between the parental populations used for the first and second cycles. Genetic diversity was greater in the UA group than in the LA+T group. The number of rare and of private alleles was reduced for both genetic groups, as well as the number of the frequent alleles in the LA+T group.     

Changes in genetic diversity during seven cycles of recurrent selection for grainyield in oat, Avena sativa L.

Variability for desirable alleles within elite breeding populations is a requirement for long-term genetic improvement. Changes in genetic diversity were evaluated in a recurrent selection programme for enhanced grain yield in oat, Avena sativa L., using coefficient of parentage (r_p) and restriction fragment length polymorphism (RFLP) based genetic similarity (S) estimates. Both r_p and s increased during seven cycles of recurrent selection, indicating a reduction in the level of genetic diversity within the population. The relationship between genetic diversity and agronomic performance was also examined. Genetic similarity measures were poor predictors of the near-homozygous progeny performance and general and specific combining ability effects for three agronomic traits in the recurrent selection programme. In addition, there was no apparent trend for preferential selection of progeny from either more similar or more diverse parents within a given cycle of selection. The overall reduction in genetic diversity in this population has not affected selection response for grain yield. Variability remaining at important loci or de novo variation are possible explanations for the continued selection progress.     

Genetic diversity among populations and breeding lines from recurrent selection in Brassica napus as revealed by RAPD markers

Recurrent selection facilitated by dominant male sterility has been conducted to broaden the genetic basis for cultivar development in Brassica napus. This study aimed to evaluate the genetic variation in four base populations (C0‐C3) and breeding lines from two of the populations produced during recurrent selection by random amplified polymorphic DNA (Rapd) markers. Genetic variation in four populations declined gradually with the advance of selection cycles as measured by expected genetic heterozygosity (from 0.2058 in C0 to 0.1536 in C3) but the decline was not statistically significant. When compared with the average genetic distances for 21 germplasm collections with wide geographical and genetic origins (0.4712) and seven breeding lines from pedigree selection (0.2059), seven breeding lines selected from the C1 population and 11 from the C3 population had a larger average genetic distance (0.5339 and 0.5486, respectively). Clustering analysis indicated that the lines from recurrent selection had a much lower genetic similarity than lines from pedigree selection. Our results suggest that base populations derived from recurrent selection could provide a wider genetic variation for selection of breeding lines with more broad genetic bases.     

Polymorphic microsatellite marker set for <Emphasis Type="Italic">Carica papaya</Emphasis> L. and its use in molecular-assisted selection

Conventional methods for the selection of papaya pure lines are time-consuming. Procedures involving the use of molecular markers and the indirect selection of homozygous plants can reduce this time considerably. The objective of this study was to evaluate the informativeness of a microsatellite marker set when used in marker-assisted selection (MAS) for the development of new papaya lines. Eighty-three lines originating from two segregating F₃ populations and from papaya germplasm were used for the molecular analysis of 27 microsatellite primers. Twenty polymorphic microsatellite primers were identified, allowing the identification of 86 alleles, with an average of 3.18 alleles per primer. The observed heterozygosity values were low for both the markers (0.00–0.29) and the individual lines (0.00–0.35). The inbreeding coefficient (f) ranged from 0.634 to 1.00. Eleven lines with f = 1.00 and 18 lines with f varying from 0.953 to 0.961 were identified. In addition, papaya lines showed high genetic diversity, which will certainly contribute to the development of new varieties. Our results show that the use of microsatellites in MAS is a quick and effective procedure for the development of papaya lines.     

Preliminary evaluation of F1 generation derived from two common bean landraces (Phaseolus vulgaris) from Nicaragua

Germplasm improvement is a conceivable solution for increasing food production in developing countries. The objectives of the present study were to incorporate the genetic diversity of two Nicaraguan common bean landraces into a segregating population and to evaluate its potential as breeding material. Adapted landraces PV0006 and PV0023 were selected as parents for single crosses based on molecular and phenotypic characteristics. The results indicate that we succeeded in reshuffling most of the genetic diversity of both parents into a segregating population. The level of allele recombinations suggests that theoretically between 128 and 1024 different pure lines could be obtained. We found putative associations between alleles and yield‐determinant components. Computer simulations demonstrated that the pedigree method and the marker selection of the most segregating individuals in the F₁ generation could improve the genetic gains for yield. These findings expose the potential of this segregating population and the feasibility of local landraces for bean improvement.     

Genetic diversity among tea cultivars from China, Japan and Kenya revealed by ISSR markers and its implication for parental selection in tea breeding programmes

Tea plant [Camellia sinensis (L.) O. Kuntze] is an important beverage crop in the world. In recent years many clonal tea cultivars have been released, and they play major roles in improving the production and quality of tea. It is important to understand the genetic diversity and relatedness of these cultivars to avoid inbreeding and narrow genetic basis in future tea breeding. In the present study, genetic diversity and relationship of 48 tea cultivars from China, Japan and Kenya were evaluated by inter‐simple sequence repeat (ISSR) markers. A total of 382 ISSR bands were scored, of which 381 (99.7%) were polymorphic. The ISSR primers showed high ability to distinguish between tea cultivars according to their high Resolving Power (R_P) with an average of 7.4. The mean of Nei’s gene diversity (H) and Shannon’s information index (I) were 0.22 and 0.35, respectively. More abundant diversity was revealed among cultivars in China than those in Japan and Kenya. Within Chinese populations, the level of diversity in east China was higher than that in other regions. The coefficient of genetic differentiation (G_ST) was 0.202, which indicates a high degree of genetic variation within populations. This result was further confirmed by analysis of molecular variance, which revealed the variance component within the populations (92.07%) was obviously larger than that among populations (7.93%). The level of gene flow (N_m) was estimated to be 2.0. This could be explained by frequent natural cross‐pollination and seed dispersal among tea populations. The pairwise similarity coefficient between the cultivars varied from 0.162 to 0.538. A dendrogram of 48 tea cultivars was constructed where all the tested cultivars were divided into two groups. Our data show that the genetic relationship among tea cultivars can be determined by the ISSR markers. This will provide valuable information to assist parental selection in current and future tea breeding programmes.     

SSR荧光检测解析224份山西玉米自交系的遗传结构与遗传关系

为利用杂种优势,对玉米自交系的遗传结构及遗传关系进行解析非常必要.本研究基于40个SSR位点的分析,对山西224份玉米自交系的遗传结构与遗传关系进行了解析.共检测出171个等位变异,平均每个位点4.28个;平均每个位点多态性信息量PIC、标记索引指数、基因多样性指数分别为0.44、2.02和0.50;有9份种质存在特异...     

Genetic diversity of oilseed Brassica napus inbred lines based on sequence-related amplified polymorphism and its relation to hybrid performance

Significant heterosis for seed yield in oilseed rape has created interest in the development of hybrid cultivars. The DNA‐based marker protocol, sequence‐related amplified polymorphism (SRAP) was used to determine genetic diversity among oilseed rape maintainer and restorer lines. This measure was used in an attempt to establish an association between genetic distance and heterosis in hybrids for various agronomic traits. A total of 118 polymorphic loci were generated by 18 SRAP primer combinations. Based on the polymorphism generated by the markers, calculated similarity index values ranged from 0.46 to 0.97. Cluster analysis grouped 10 maintainer and 12 restorer lines into three groups, with the exception of two maintainer lines, PM5 and PM9, which fell outside these groups. The grouping of the lines was largely in agreement with the available pedigree data on their origin and agronomic performance. Analysis of variance among inbred lines and their resulting F₁ hybrids over two locations revealed significant differences for plant height, days to maturity and seed yield, but not for oil content. Substantial mid‐parent heterosis was observed only for seed yield, and ranged from 26% to 169%. All hybrids surpassed their respective inbred lines for this trait, except for a single cross combination of related lines. In general, crosses of lines located in different clusters yielded more than those from the same clusters. Regression analysis revealed a statistically significant relationship between the genetic distance of the parents and seed yield in their hybrid, and their derived mid‐parent and high‐parent heterosis. The correlation coefficient between genetic distance and yield (0.64) indicated a moderately strong relationship, so it is possible that some of the SRAP markers might be linked to quantitative trait loci for seed yield.     

Genetic architecture of plant height in maize phenotype‐selected introgression families

This study aimed at developing, characterizing and evaluating two maize phenotypic‐selected introgression libraries for a collection of dominant plant height (PHT)‐increasing alleles by introgressing donor chromosome segments (DCS) from Germplasm Enhancement of Maize (GEM) accessions into elite inbred lines: PHB47 and PHZ51. Different backcross generations (BC₁‐BC₄) were developed and the tallest 23 phenotype‐selected introgression families (PIFs) from each introgression library (PHB47 or PHZ51) were selected for single nucleotide polymorphism genotyping to localize DCS underlying PHT. The result shows that most PIFs carrying DCS were significantly (α = 0.01) taller than the respective recurrent parent. In addition, they contained larger donor genome proportions than expected in the absence of selection or random mating across all BC generations. The DCS were distributed over the whole genome, indicating a complex genetic nature underlying PHT. We conclude that our PIFs are enriched for favourable PHT‐increasing alleles. These two libraries offer opportunities for future PHT gene isolation and allele characterization and for breeding purposes, such as novel cultivars for biofuel production.     

陕西省野生大豆种质资源的SSR遗传多样性研究

为了研究陕西地区野生大豆的遗传多样性特点,利用SSR分子标记分析了陕西省6个野生大豆(Glycine soja)天然种群和1个栽培大豆(Glycine max)种群的遗传结构与遗传多样性。结果显示：13个位点共检测出113个等位基因,平均每个位点的等位基因数(A)为8.69个,等位基因数目范围为4~13个,有效等位基因数(N_e)范围为2.135(Satt590)~9.385 (Satt487),平均有效等因基因数为5.623;观察杂合度(H_o)变化范围为0.033~0.121,平均为0.080;预期杂合度(H_e)的变化范围为0.312~0.658,平均为0.482;种群平均Shannon遗传多样性指数(I)为0.657;野生大豆种群基因多样度比率(F_ST)为0.465。该研究显示,陕西省野生大豆具有较高水平的遗传多样性,野生大豆的遗传多样性普遍高于栽培大豆;随着海拔的不断升高,野生大豆遗传多样性变低;陕西中部、南部的野生大豆种质资源丰富、种群具有较高的遗传多样性,推测该区域为陕西省野生大豆的遗传多样性中心。     

Validation of a major QTL for scab resistance with SSR markers and use of marker-assisted selection in wheat

The objectives of this study were to validate the major quantitative trait locus (QTL) for scab resistance on the short arm of chromosome 3B in bread wheat and to isolate near‐isogenic lines for this QTL using marker‐assisted selection (MAS). Two resistant by susceptible populations, both using ‘Ning7840’ as the source of resistance, were developed to examine the effect of the 3BS QTL in different genetic backgrounds. Data for scab resistance and simple sequence repeat (SSR) markers linked to the resistance QTL were analyzed in the F_2:3 lines of one population and in the F_3:4 lines of the other. Markers linked to the major QTL on chromosome 3BS in the original mapping population (‘Ning7840’/‘Clark’) were closely associated with scab resistance in both validation populations. Marker‐assisted selection for the QTL with the SSR markers combined with phenotypic selection was more effective than selection based solely on phenotypic evaluation in early generations. Marker‐assisted selection of the major QTL during the seedling stage plus phenotypic selection after flowering effectively identified scab resistant lines in this experiment. Near‐isogenic lines for this 3BS QTL were isolated from the F₆ generation of the cross ‘Ning7840’/‘IL89‐7978’ based on two flanking SSR markers, Xgwm389 and Xbarc147. Based on these results, MAS for the major scab resistance QTL can improve selection efficiency and may facilitate stacking of scab resistance genes from different sources.     

Identification of salt tolerance‐improving quantitative trait loci alleles from a salt‐susceptible rice breeding line by introgression breeding

To improve salt tolerance of two elite rice varieties, Ce258 and Zhongguangxiang1 (ZGX1), two sets of introgression lines (ILs) each comprising 200 BC₁F₁₀ lines derived from a common donor, IR75862, and two recipient parents, Ce258 and ZGX1, were used for mapping of QTLs for four salt tolerance‐related traits at the seedling stage. Although the three parents were susceptible to salt, the two IL populations showed transgressive segregations for salt tolerance with 12 and 8 salt tolerance ILs in the Ce258‐ILs and ZGX1‐ILs. Eighteen main‐effect QTLs were identified for the four traits in the two IL populations, and the IR75862 alleles at most loci showed increased and decreased salt tolerance in the ZGX1 and Ce258 backgrounds, suggesting overwhelming genetic background effects on QTL detection for salt tolerance. The qDSS11 simultaneously detected in the two backgrounds was validated in a F₂ population derived from a salt tolerance line and ZGX1. Our results indicated that salt tolerance‐enhancing allele could be identified in the elite susceptible breeding lines and that introgression of the favourable alleles could facilitate the development of superior lines with greater salt tolerance levels.     

Detecting major QTL associated with resistance to bacterial blight using a set of rice reciprocal introgression lines with high density SNP markers

Bacterial blight (BB), caused by Xanthomonas oryzae pv. oryzae (Xoo), is a worldwide rice disease. QTL providing BB resistance were identified using a set of introgression lines (ILs) derived from a cross between Teqing and Lemont, and 3924 evenly distributed SNP markers developed from the two parents. After inoculating three Xoo pathotypes, CII, CIV and CV, seven major BB resistance‐associated QTL were detected. The alleles at all loci with improved BB resistance were from the Teqing background. Four QTL resistance to CII and CIV were identified in the reciprocal backgrounds across 2 years. Six QTL conferring resistance to CV identified in Teqing background were not detected in Lemont background, indicating that genetic background had a strong effect on the BB resistance‐associated QTL. Based on the interactions of 27 significant digenic QTL pairs among the seven main‐effect QTL, the QTL were divided into three strain‐specific groups. Genotype analyses of resistant ILs suggest that rice lines with a high level of resistance to BB can be achieved by pyramiding R gene(s) and QTL that interact with R gene(s).     

Improvement of resistance to Fusarium head blight by recurrent selection in an intermating breeding spring wheat population using the dominant male-sterile gene ms 2

Four cycles of recurrent selection for FHB resistance were conducted in an intermating wheat breeding population using the dominant male-sterile gene ms ₂ during 1987–1991.Five cycles of phenotypic mass selection for male-sterile plants were evaluated using the soil-surface inoculation method in Experiment I. Experiment II evaluated changes in FHB scores during five cycles of progeny selection for fertile plants using the single-floret inoculation method. In Experiment I, the average level of FHB response increased to MR level in C₄, compared to MS level in C₀. The numbers of infected spikelets and diseased kernels decreased 0.32 and 2.68 per cycle, respectively. In Experiment II, the average level of FHB response increased to R level in C₄F₁. The numbers of infected spikelets and diseased kernels decreased 0.93 and 4.58 per cycle, respectively. In both experiments, the largest selection gains were realized in the first cycle. The frequencies of R and MR individuals were increased significantly. The frequencies of individuals with FHB response equal and/or superior to Sumai 3 were increased to 5–8% in C₄ and 25% in C₄F₁after the fourth cycle. Agronomic traits tended to be slightly improved in selected populations. Compared to 2% in C₀, about 34% of lines superior in both FHB resistance and agronomic traits in C₄F₁ were selected to enter the conventional breeding program for further evaluation. Sixty three semidwarf lines superior in both FHB resistance and yield potential were selected from the F₅ generations derived from C₁F₁ to C₄F₁. From them, two resistant cultivars with high-yielding potential were developed and commercialized in the Lower Yangtze Valley. Recurrent selection appears to be highly effective and feasible in shifting the average FHB response of the intermating population in the desirable direction, thereby enhancing the frequency of resistant individuals. This revised version was published online in July 2006 with corrections to the Cover Date.     

Temporal genetic structure patterns in tropical maize populations under reciprocal recurrent selection

In order to investigate the effect of long term recurrent selection on the pattern of gene diversity, thirty randomly-selected individuals from the progenitors (p) and four selection cycles (C0, C3, C6 and C11) were sampled for DNA analysis from the tropical maize (Zea mays L.) breeding populations, Atherton 1 (AT1) and Atherton 2 (AT2). Fifteen polymorphic Simple Sequence Repeat markers amplified a total of 284 and 257 alleles in AT1 and AT2 populations, respectively. Reductions in the number of alleles were observed at advanced selection cycles. About 11 and 12% of the alleles in AT1 and AT2 populations respectively, were near to fixation. However, a higher number of alleles (37% in AT1 and 33% in AT2) were close to extinction. Fisher’s exact test and analysis of molecular variance (AMOVA) showed significant population differentiations. Gene diversity estimates and AMOVA revealed increased genetic differentiations at the expense of loss of heterozygosity. Population differentiations were mainly due to fixation of complementary alleles at a locus in the two breeding populations. The estimates of effective population at an advanced selection cycle were close to the population size predicted by the breeding method.     

Identification of cultivars and accessions of Lolium, Festuca and Festulolium hybrids through the detection of simple sequence repeat polymorphism

Molecular diversity and genetic affinity in the Lolium/Festuca grass complex have been assessed using simple sequence repeat (SSR) marker technology. The genotypic set was derived from three accessions of perennial ryegrass, two cultivars of Italian ryegrass, two cultivars of meadow fescue, two cultivars of tall fescue and 10 accessions from different intergeneric hybrid (Festulolium) combinations. The majority of the genomic DNA‐derived SSR primer pairs from perennial ryegrass (LPSSR) and Italian ryegrass (LMSSR) produced clear, simple and distinctive amplification products from the majority of the genotypes. The efficiency of cross‐specific amplification for LPSSR markers varied from 38% in meadow fescue to 93% in two cultivars of Festulolium and from 57% in meadow fescue to 87% in Italian ryegrass for LMSSR markers. Of 40 amplified markers, 14 (35%) produced species‐difference alleles in the relation to cultivars used in the present study. Thirty‐five LPSSR locus‐derived alleles were found to be specific to Lolium species, four to meadow fescue and six to tall fescue. For LMSSR alleles, eight were specific to Lolium species and five were only associated with Italian ryegrass, and null alleles were detected for meadow fescue in all instances. These species‐difference markers could clearly identify different accessions of Festulolium. Cluster analysis separated the individual taxa and showed grouping of intergeneric hybrids based on genomic composition. The data distinguished between the species and reflected the known pedigree of the cultivars and the differences between the species. The dendrogram also distinguished between the Festulolium accessions and clearly demonstrated the relations between Festulolium hybrids and their parent species.