Genotypic variation of quantitative trait loci controlling in vitro androgenesis in maize.

A quantitative trait loci (QTL) analysis for androgenetic capability has been conducted on three different crosses in maize, including very high and nonresponding lines for androgenesis. The doubled haploid lines derived by anther culture from the crosses DH5 x DH7, A188 x DH7, and R6 x DH99 showed a range of 0-70%, 0-40%, and 0-50% androgenetic responding anthers, respectively. The genotypic heritability of means for this trait is close to 0.90 for A188 x DH7 and 0.78 for R6 x DH99. The QTL analysis involved in each population the mapping of more than 100 loci covering a large part of the genome with reasonably spaced markers averaging 12 cM. Different measurements describing the androgenetic process were studied: AC, percentage of responding anthers; ELS, number of androgenetic embryos produced per 100 plated anthers; PLE, number of plantlets regenerated per 100 embryos; PLA, number of plantlets per 100 plated anthers. In each cross, three to four QTLs were found for AC, explaining 30-40% of the phenotypic variation. The QTL detected for PLA was also strong QTL for AC or ELS. This agrees with the observation that these last two traits are good predictors for final plantlet yield. The QTLs found were specific, although the same line DH7 was used in two crosses and DH99 derived from DH5 and DH7 in the third cross. These results suggest that the transfer of the androgenetic capabilities in elite germplasm will still involve a phenotypic evaluation of the androgenetic performances. A backcross-assisted selection based only on the genotype at the QTL is probably possible but only within the crosses used for this QTL analysis.     

籼稻花粉无性系变异的研究

对７个籼稻品种通过花药培养获得的１６１个花粉植株进行了花粉无性系变异的研究。结果表明：（１）所有花粉无性系在遗传上都是纯合的,其整齐度与起始亲本相近或超过起始亲本;（２）花粉无性系的变异主要在数量性状上有一定变幅,变异方向有负向也有正向的。同一来源的无性系间的变异系数略高于起始亲本,仅千粒重达到显著标准。以所考查的性状为基数,变异的频率为１２．５％,同时有３－４个性状发生变异的无性系数占总数的４．３％;（３）少数变异体性状有重大变异,并具有育种价值;（４）根据１０种同工酶和ＲＦＬＰ分析,仅从３个性状发生重大变化的变异体中检测出与起始亲本的差异。     

A comparison of androgenetic doubled-haploid,and single seed descent lines in Triticale

Summary Sixty single seed descent (SSD) lines and about 25 anther-derived doubled-haploid (DH) lines were obtained from two triticale crosses. The frequency distributions of 10 quantitative agronomic traits were compared using parametric and non-parametric tests. A multivariate discriminant analysis was subsequently carried out. Gliadin patterns obtained from each line by polyacrylamide gel electrophoresis were used to calculate intra- and inter-population diversities from relative dissimilarity indices. It was found that DH and SSD lines show significant differences in frequency distributions of 1000 grain weight in both crosses, of heading date for one cross, and of lodging susceptibility for the other cross. The results of intra- and inter-population gliadin diversity indicate that although the SSD method theoretically provides more opportunity for recombination to occur than the DH method, it did not produce a greater range of recombinants. Since there is no significant difference between SSD- and DH-line distributions for grain yield, anther culture appears to be an efficient method for producing high yielding homozygous lines from F₁ hybrids of triticale in a relatively short time.     

Comparison of single seed descent and anther culture-derived lines of three single crosses of rice

Summary Distribution parameters (mean, variance, skewness and kurtosis) of 12 quantitative traits were evaluated for inbred lines generated through single seed descent (SSD) and anther culture (AC) of two japonica x japonica hybrids and one japonica x indica hybrid of rice. For most of the traits the data were normally distributed, and the means and variances were found to be identical for SSD- and AC-derived lines of the given hybrids. However, for some other traits, differences between the two population types were observed, mainly in the lines derived from the intra-japonica crosses. A tentative explanation for these differences is given. Nevertheless, our results suggest that SSD and AC are equally effective breeding techniques for producing agronomically useful lines of rice.     

Numerical method for detection of linkage between genes for two metrical traits

A homozygous population derived from hybrids between two homozygous parents may be used for genetic analysis of metrical traits. The paper describes the use of doubled haploids (DH) and single seed descent (SSD) lines for detection of linkage between genes conditioning two quantitative traits. A computational algorithm is presented, which facilitates matching various variants of relations between variances, covariances and means of DH and SSD populations so as to make it possible to conclude on the presence/absence of linkage. The suggested methodology is illustrated with an example concerning three quantitative traits of barley: length of the third internode, stem wall thickness, and 1000-grain weight.     

Comparison of frequency distributions of doubled haploid and single seed descent lines in barley

Summary Both doubled haploid (DH) and single seed descent (SSD) methods were used to derive homozygous lines from two crosses of barley. The frequency distributions of grain yield, heading date, and plant height of the DH and SSD lines were compared by the Mann-Whitney U test, Kolmogorov-Smirnov twosample test and Wald-Wolfowitz runs test. It was found that the DH lines distributed in the same manner as the SSD lines with respect to the three characters. The results indicated that although the SSD method had more opportunity for recombination than the DH method, it did not produce a sample of recombinants which differed significantly from the DH sample; thus both methods were equally efficient for use in deriving homozygous lines from F₁ hybrids in a relatively short time.Contribution no. 455 Charlottetown Research Station, Agriculture Canada, P.E.I. (Canada)     

Comparison of bread wheat lines selected by doubled haploid, single-seed descent and pedigree selection methods

 Yield performance of each group of ten spring bread wheat lines selected by doubled haploid (DH), single-seed descent (SSD) and pedigree selection (PS) methods from three F₁ crosses was compared with the aim of evaluating the DH method in breeding programs. Populations of 65–97 DH lines and 110 SSD lines per cross were used for selection. PS lines were developed by repeated selections from 1500 F₂ plants. Yield evaluation was performed at the F₆ generation of SSD and PS lines along with DH lines in a 2-year field experiment. It took only 2 years from the planting of wheat materials for DH production to the planting of selected DH lines for yield evaluation. There was no significant difference in grain yield between DH lines and PS lines selected from an F₁ cross whose parental varieties were closely related in their pedigrees. In two crosses with low coefficients of parentage and a large variation in their progenies, grain yield of selected DH lines was significantly lower than those of selected SSD and PS lines. These results confirm that the DH method can save time in obtaining recombinant inbred lines ready for yield evaluation. However, a larger DH population is required to achieve the same level of genetic advance with the PS method in crosses containing greater genetic variation. Received: 23 December 1997 / Accepted: 12 March 1998     

Variation of DNA fingerprints among accessions within maize inbred lines and implications for identification of essentially derived varieties.

Genetic distances (GDs) based on molecular markers are important parameters for identifying essentially derived varieties (EDVs). In this context information about the variability of molecular markers within maize inbred lines is essential. Our objectives were to (1) determine the variation in the size of simple sequence repeat (SSR) fragments among different accessions of maize inbreds and doubled haploid (DH) lines, (2) attribute the observed variation to genetic and marker system-specific sources, and (3) investigate the effect of SSR fragment size differences within maize lines on the GD between maize lines and their consequences for the identification of essentially derived varieties. Two to five accessions from nine inbred lines and five DH lines were taken from different sources or drawn as independent samples from the same seed lot. Each accession was genotyped with 100 SSR markers that evenly covered the whole maize genome. In total, 437 SSR fragments were identified, with a mean of 4.4 alleles per locus. The average polymorphic information content (PIC) was 0.58. GD estimates between two accessions of the same genotype ranged from 0.00 to 0.12 with an average of 0.029 for inbred lines and 0.001 for DH lines. An average of 11.1 SSRs was polymorphic between accessions of the same inbred line due to non-amplification (8.1 SSRs), heterogeneity (4.0 SSRs) or unknown alleles (2.6 SSRs). In contrast to lab errors, heterogeneity contributed considerably to the observed variation for GD. In order to decrease the probability to be suited for infringing an EDV threshold by chance, we recommend to increase the level of homogeneity of inbred lines before applying for plant variety protection.     

A comparison of inbred lines derived by doubled haploidy and single seed descent in spring barley (Hordeum vulgare)

Samples of F_x inbred lines, derived by doubled haploidy (DH) and single seed descent (SSD) from five spring barley crosses were compared for agronomic characters. It was shown that, over this range of diverse crosses, inbreds derived by either technique could surpass the better parent or even the heterotic F₁. The means of the DH and SSD were, however, different for a number of characters, as well as differing from the mid-parent value. It was concluded that these differences stemmed from the presence of interacting genes showing linkage disequilibrium, although there was no unambiguous test to distinguish this from differential survival during the production of inbreds. This view was further supported by the finding that the DH sample, which tends to preserve existing linkages, produced a higher proportion of lines exceeding the better scoring parent when compared with the SSD population.     

Structural organization of the group-1 chromosomes of two bread wheat sister lines

Eureka and Renan are two French bread wheat cultivars derived from a 4-way cross. Using molecular markers (essentially RFLPs), we studied the structure of the group-1 chromosomes of these two genotypes, their parents and a doubled-haploid (DH) population derived from their F(1). Using the DH population (102 lines), a molecular map of the three homoeologous group-1 chromosomes was produced and compared with the map established on another intervarietal cross: Courtot x Chinese Spring (Cadalen et al. 1997). The polymorphic markers were mapped on the DH population and characterized on the four grand-parents, allowing us to compare the structural organization of the group-1 chromosomes of Eureka and Renan and determine their origin. These chromosomes were very different, except for small regions (1AL proximal and 1BL distal) which were identical.     

The agronomic performance of anther culture derived plants of barley produced via pollen embryogenesis

Anther culture was used to generate microspore-derived doubled haploid (DH) plants from four spring barley crosses. The culture medium used contained maltose as the sole carbohydrate source and the mode of plantlet regeneration was mainly via pollen embryogenesis. Both haploid and spontaneously doubled regenerants were produced and the doubled haploids were compared to recom-binant inbred lines generated by several rounds of selfing (single seed descent). Parental, DH and single seed descent (SSD) lines were grown in randomised, replicated field trials and the samples were scored for a range of agronomic traits. The mean performance and phenotypic distribution of the DH and SSD samples were similar and there was little evidence to support the conclusion that anther culture derived lines exhibit a reduction in vigour. Where significant differences were detected between groups these were mainly confined to crosses which were segregating for the denso dwarfing gene. The differential transmission of particular regions of the barley genome may therefore influence and confound the expression of agronomic traits in DH populations. This is the first report of the agronomic performance of anther culture lines produced via pollen embryogenesis and the results are discussed in relation to the exploitation of anther culture technology in barley breeding.     

The agronomic performance of wheat doubled haploid lines derived from wheat x maize crosses

Summary The agronomic performance of 9 doubled haploid (DH) lines of Chinese Spring, 6 DH lines of Hope, 14 DH lines of the single chromosome substitution line Chinese Spring (Hope 5 A) and their respective parents was analyzed under field conditions. Seventeen Chinese Spring DH lines derived from wheat x Hordeum bulbosum crosses were also included for comparison. No significant variation was detected in either population of Chinese Spring DH lines and neither DH population differed from its parent. The Hope DH lines differed significantly for tiller biomass, spikelet number per ear, ear grain weight and 50-grain weight. However, all the variation could be attributed to the poor performance of only one line. Chinese Spring (Hope 5 A) DH lines showed significant variation for ear emergence time, but this was probably due to genetic heterogeneity in the parental stock. Overall, the results suggest that most DH lines produced by the wheat x maize method resemble their wheat parent, and that the variation induced in DH production is likely to be similar to that found in DHs from wheat x Hordeum bulbosum crosses.     

Variation of DNA fingerprints among accessions within maize inbred lines and implications for identification of essentially derived varieties: II. Genetic and technical sources of variation in AFLP data and comparison with SSR data

Accuracy and reproducibility of genetic distances (GDs) based on molecular markers are crucial issues for identification of essentially derived varieties (EDVs). Our objectives were to investigate (1) the amount of variation for amplified fragment length polymorphism (AFLP) markers found among different accessions within maize inbreds and doubled haploid (DH) lines, (2) the proportion attributable to genetic and technical components and marker system specific sources, (3) its effect on GDs between maize lines and implications for identification of EDVs, and (4) the comparison to published SSR data from the same plant materials. Two to five accessions from nine inbred lines and five DH lines were taken from different sources of maintenance breeding or drawn as independent samples from the same seed lot. Each of the 41 accessions was genotyped with 20 AFLP primer combinations revealing 988 AFLP markers. Map positions were available for 605 AFLPs covering all maize chromosomes. On average, six (0.6%) AFLP bands were polymorphic between different accessions of the same line. GDs between two accessions of the same line averaged 0.013 for inbreds and 0.006 for DH lines. The correlation of GDs based on AFLPs and SSRs was tight (r = 0.97**) across all 946 pairs of accessions but decreased (r = 0.55**) for 43 pairs of accessions originating from the same line. On the basis of our results, we recommend specific EDV thresholds for marker systems with different degree of polymorphism. In addition, precautions should be taken to warrant a high level of homogeneity for DNA markers within maize lines before applying for plant variety protection.     

Genetic evaluation of haploid clonal lines of a single donor plant of Populus maximowiczii

Summary Haploid clonal lines derived from anthers of a single donor plant of Populus maximowiczii were evaluated for several quantitative traits. In a nursery test, variances due to clonal lines ranged from 8% to 12% of the phenotypic variance in growth cessation and flushing date, respectively. No variance in relative shoot growth rate was associated with clonal lines. In a greenhouse study, gametoclonal variance in several leaf morphology traits ranged from 9% to 37% of the total variance. In relative wood density, variation due to haploid lines accounted for 25% of the total variance. In an isozyme analysis of 20 haploid and dihaploid plants, significant non-Mendelian segregation in isocitrate dehydrogenase was detected. The implications of these results for tree breeding are discussed.     

水稻双单倍体群体的分子标记图示基因型分析

利用窄叶青８号（籼稻）／京系１７（粳稻）花培产生的双单倍体群体建立了一个包含１６０个分子标记的遗传连锁图,在此基础上利用ＨＹＰＥＲＧＥＮＥ软件建立了５２个ＤＨ系的图示基因型,并对ＤＨ系的亲本基因组比率和染色体的交换重组频率进行了比较分析。结果表明本实验所用的ＤＨ群体没有显著偏离正态分布,籼粳稻杂交后代中植株的籼粳表现与同工酶、形态指数和基因组比率的分析结果一致,此外还发现ＤＨ群体中出现了大量的交换罕见染色体。利用图示基因型分析发现株高和分子标记ＲＺ９７８和ＲＧ４Ａ相关,生育期和ＲＲＫ０８－１、ＲＧ４７７和ＲＧ５１１相关。本文还就图示基因型分析技术在ＤＨ群体的遗传分析和选择育种中的应用进行了讨论．     

Use of a subset of doubled-haploid lines for RAPD interval mapping in barley.

Molecular markers have been used in barley to locate genes and quantitative trait loci. Only a few RAPD markers have been located on barley marker maps. The objectives of this study were (i) to place RAPD markers in specific intervals on the barley linkage map developed from the cross Steptoe (S) x Morex (M), (ii) to examine the distribution of RAPD markers, and (iii) to compare markers amplified by Taq DNA polymerase with those amplified by the Stoffel fragment of Taq DNA polymerase. Screening of DNA from S and M with 362 decamer primers identified 85 that amplified 127 reliable RAPDs. A subset of 15 doubled-haploid (DH) lines from the 150 DH line mapping population was used to place these RAPD markers in intervals on the SM map. This subset can be used for rapid placement of any new markers on the SM linkage map. Most of the RAPD markers were dominant but four codominant RAPDs were identified. The RAPDs were not evenly distributed, with many clustered around the centromeric region of each chromosome. Two of these clusters were located in intervals larger than 15 cM. Testing of 38 to 42 additional DH lines provided more precise placement of eight of the markers in these clusters. Reliable RAPDs were detected with 44% of the primers tested with the Stoffel fragment, but with only 17% of the primers tested with Taq DNA polymerase. These RAPDs provide additional markers for use in barley improvement.     

QTL and QTL x environment effects on agronomic and nitrogen acquisition traits in rice

Agricultural environments deteriorate due to excess nitrogen application.Breeding for low nitrogen responsive genotypes can reduce soil nitrogen input.Rice genotypes respond variably to soil available nitrogen.The present study attempted quantification of genotype x nitrogen level interaction and mapping of quantitative trait loci (QTLs) associated with nitrogen use efficiency (NUE) and other associated agronomic traits.Twelve parameters were observed across a set of 82 double haploid (DH) lines derived from IR64/Azucena.Three nitrogen regimes namely,native (0 kg/ha; no nitrogen applied),optimum (100 kg/ha) and high (200 kg/ha) replicated thrice were the environments.The parents and DH lines were significantly varying for all traits under different nitrogen regimes.All traits except plant height recorded significant genotype x environment interaction.Individual plant yield was positively correlated with nitrogen use efficiency and nitrogen uptake.Sixteen QTLs were detected by composite interval mapping.Eleven QTLs showed significant QTL x environment interactions.On chromosome 3,seven QTLs were detected associated with nitrogen use,plant yield and associated traits.A QTL region between markers RZ678,RZ574 and RZ284 was associated with nitrogen use and yield.This chromosomal region was enriched with expressed gene sequences of known key nitrogen assimilation genes.     

籼粳杂种双单倍体的配子选择

对典型的灿与粳稻杂种,窄叶青８号／京系１７Ｆ１花药进行培养获得的１３２个双单倍体的形态特性、同工酶与ＲＦＬＰ标记的分离与重组进行了考察分析,研究是否存在配子选择问题。结果表明：（１）对４个重要数量性状和６个涉及籼、粳特征的形态指数进行考察所获数据均为连续分布,并呈正态曲线;（２）用８种同工酶对５２个ＤＨ系分析结果表明,只有２种同工酶显著偏离期望的１：１比率,而灿与粳的总基因型比率相近;（３）应用１６７个ＲＦＬＰ标记对１３２个ＤＨ系进行的分析发现,有３６％标记发生偏分离,但偏籼与偏粳的比率相近,两个亲本基因组在ＤＨ群体中所占比率相同（各５０％）,各种基因组成呈正态分布。综上所述,本研究虽观察到一些轻微偏分离现象,但籼粳基因基本上随机分离与重组,等位基因总频率未偏离１：１比率。     

Comparative mapping of the Oregon Wolfe Barley using doubled haploid lines derived from female and male gametes

The Oregon Wolfe Barley mapping population is a resource for genetics research and instruction. Prior reports are based on a population of doubled haploid (DH) lines developed by the Hordeum bulbosum (H.b.) method, which samples female gametes. We developed new DH lines from the same cross using anther culture (A.C.), which samples male gametes. Linkage maps were generated in each of the two subpopulations using the same 1,328 single nucleotide polymorphism markers. The linkage maps based on DH lines derived from the products of megasporogeneis and microsporogenesis revealed minor differences in terms of estimated recombination rates. There were no differences in locus ordering. There was greater segregation distortion in the A.C.-derived subpopulation than in the H.b.-derived subpopulation, but in the region showing the greatest distortion, the cause was more likely allelic variation at the ZEO1 plant height locus rather than to DH production method. The effects of segregation distortion and pleiotropy had greater impacts on estimates of quantitative trait locus effect than population size for reproductive fitness traits assayed under greenhouse conditions. The Oregon Wolfe Barley (OWB) population and data are community resources. Seed is available from three distribution centers located in North America, Europe, and Asia. Details on ordering seed sets, as well as complete genotype and phenotype data files, are available at http://wheat.pw.usda.gov/ggpages/maps/OWB/.