Production of Gynogenetic Plants of Onion (Allium cepa L.) after Crossing with Irradiated Pollen

The possibility of producing gynogenetic plants with pollen inactivated by irradiation was explored for the onion (Allium cepa L.). Two successive experiments included the use of cytoplasmic male sterility, recessive nuclear markers in the female plants and pollen irradiation of 150 Gy. Pollinations were achieved with the aid of flies and all seeds were harvested without the use of embryo rescue. Plantlets with recessive characters could be detected and they contained haploid and diploid cells. The development of these plantlets until flowering was observed under agronomic conditions. The use of doubled haploid (DH) plants in onion breeding is discussed.     

Molecular mapping of QTLs for non-parasitic leaf spot resistance and comparison of half-sib DH populations in spring barley

Quantitative trait loci (QTLs) for resistance against non-parasitic leaf spots (NPLS) were first characterized in a spring barley double haploid population derived from the cross IPZ 24727/Barke (Behn et al., 2004). The aim of the present study was to identify QTLs for NPLS resistance in the half-sibling DH population IPZ 24727/Krona and to compare them with the QTLs of the population IPZ 24727/Barke. An anther culture-derived doubled haploid population of 536 DH lines was developed from the cross IPZ 24727 (resistant)/Krona (susceptible). Field trials were performed over two years in two replications, scoring NPLS and agronomic traits that might interact with NPLS. A molecular linkage map of 1035 cM was constructed based on AFLPs, SSRs and the mlo marker. QTL analyses for NPLS identified three QTLs that accounted for 30% of the phenotypic variation. For comparison of the QTLs from each DH population, a consensus map was generated comprising 277 markers with a length of 1199 cM. In both populations, the QTLs for NPLS mapped to chromosomes 1H, 4H and 7H. A common QTL with a great effect in both populations and over all environments was localized at the mlo locus on chromosome 4H, indicating that the mlo powdery mildew resistance locus has a considerable effect on NPLS susceptibility. The steps necessary to validate the QTLs and to improve the NPLS resistance by breeding were discussed.     

Parental effects on the performance of cultivated × wild species hybrids in potato

Valuable genetic diversity in diploid wild Solanum species can be accessed through crosses to haploids (2n = 2×) of the tetraploid cultivated potato, Solanum tuberosum. Haploid-wild species hybrids segregate for the ability to tuberize in the field. In addition, they vary in male fertility, vine size, stolon length, and tuber size. In this study, three haploids were crossed with nine diploid wild Solanum species and 27 hybrid families were evaluated in the field for two years. The proportion of male fertile hybrid clones varied depending on the wild species parent. A large effect of the female parent was detected for vine size, stolon length, tuber size, percent tuberization, and percent plants selected for agronomic quality. An exceptional haploid (US-W4) was identified for the production of agronomically desirable haploid-wild species hybrids. In hybrids derived from US-W4, differences among wild species parents were observed for agronomic quality. Superior hybrids were produced by S. berthaultii and S. microdontum. Reciprocal crosses were evaluated for a subset of families. When the wild species was used as the female parent, male fertility was restored, but tuberization and tuber size were reduced. Careful selection of both haploid and wild species parents can result in a large proportion of fertile, agronomically desirable hybrid offspring.     

Associations between three ml-o powdery mildew resistance genes and agronomic traits in barley

Summary A population of 198 chromosome-doubled haploid lines of spring barley was scored for segregation in locus ml-o (powdery mildew reaction) on chromosome 4 and in the linked loci s (rachilla hair length) and ddt (reaction to the insecticide DDT) on chromosome 7. They were also tested in a disease-free field trial for the agronomic traits: grain yield, thousand grain weight, lodging, and necrotic leaf spotting. The three mutagen-induced resistance genes ml-o5, ml-o6 (from Carlsberg II) and ml-10 (from Foma) showed no detectable differences with respect to effects on agronomic traits. They all conferred a four per cent reduction in grain yield caused mainly by lower thousand grain weight, and an increase in necrotic leaf spotting. The two original mutants of Carlsberg II had additional mutant genes affecting agronomic traits. Lines with gene S (long hair) had on average a three per cent higher thousand grain weight than those with s. The alleles in locus ddt showed no association with the agronomic traits. It is concluded i) that the associations between the three ml-o alleles and agronomic traits are caused by pleiotropy, ii) that ml-o resistant, high-yielding lines may be selected, and iii) that the association between gene s and thousand grain weight may be due to genetic linkage.Abbreviations DH-lines chromosome-doubled haploid lines     

Doubled haploids of Coffea canephora: development,fertility and agronomic characteristics

Summary Doubled haploids (DH) of Coffea canephora Pierre were developed using haploid embryos which occur spontaneously in association with polyembryony. The frequencies of polyembryonic seeds and haploid embryos varied according to the parental genotypes. However, production of a large number of DH seemed possible from all genotypes. More than 750 DHs produced from various genotypes were grown under field conditions and evaluated for different characters of agronomic importance. Approximately half of DH genotypes did not survive, suggesting a strong, negative effect of homozygosity. Inbreeding depression is particularly severe on general vigor and reproductive aspects. For several characters studied such as leaf shape, leaf rust resistance and hundred bean weight, considerable genetic variations were observed within and between groups of DHs constituted by the DHs produced from the same clone. Despite their low vigor and reduced fertility, the DHs of C. canephora offer new possibilities in genetic research and coffee breeding.     

Development of in vivo haploid inducers for tropical maize breeding programs

Lack of adapted haploid inducers currently impedes adoption of the doubled haploid technology in tropical maize breeding programs. Our objective was to generate inducers with improved adaptation to tropical conditions. We developed segregating generations from crosses between temperate inducers having haploid induction rates (HIR) of 8–10 % and tropical CIMMYT maize lines (CML; HIR = 0 %) and evaluated these for HIR and agronomic performance under tropical lowland field conditions. The applied pedigree breeding scheme comprising mass selection on individual F₂ plants for highly heritable and visually scorable traits, followed by family-based selection for HIR and other agronomic characteristics in advanced selfing and backcross (BC) generations seems highly suitable for breeding improved haploid inducers with adaptation to different agroecologies. The most advanced tropical inducer candidates (TIC) combine HIR of up to 10 % with improved pollen production, disease resistance, and plant vigor compared to temperate inducers under tropical conditions. Agronomic characteristics were significantly improved in the BC to CML compared to BC to inducers, while mean HIR of both populations were similar, indicating that backcrossing to the adapted parent was suitable to improve adaptation of new inducers without sacrificing high HIR. When screening random open-pollinated maize accessions, HIR of up to 3 % were observed, suggesting that novel genetic variation may be present in maize accessions that could be exploited to improve HIR in maize. In conclusion, tropical inducer development proceeds well, but evaluation of TIC in multi-environment trials needs to be completed before large-scale dissemination can commence.     

Inheritance and potentials of a mutated dwarfing gene ndf1 in Brassica napus

A dwarf mutant ‘NDF‐1′, approximately 70 cm high, was derived from a 200‐cm high doubled haploid (DH) line ‘3529’ (Brassica napus), seeds of which were jointly treated with chemical inducers and bombardment of fast neutron. The leaves of the ‘NDF‐1’ mutant were wrinkled and thicker compared with the wild‐type control. The mutant had much lower values than its original parents for all agronomic traits, except for its seed weight. A genetic analysis revealed that dwarfism is under the control of a major gene (designated as ndf1) with a mainly additive effect and non‐significant dominance effect. Because of the high level of resistance to lodging, breeding programmes for double low dwarf oilseed rape and heterosis utilization were initiated. Some new dwarf strains with improved agronomic performance were developed. The hybrid of the cross between the tall parent and the dwarf line showed increased harvest index and significantly higher seed yield than the tall parent or the control variety ‘Zhongyou 821’ and presented an estimated heterosis vigour rate as high as 12.5–25.8%. The dwarf trait will be a promising marker for a simple, economic and efficient way to control the purity of F₁ hybrid varieties in hybrid production of B. napus.     

Comparison and analysis of main effects,epistatic effects,and QTL × environment interactions of QTLs for agronomic traits using DH and RILs populations in rice

Two genetic linkage maps based on doubled haploid (DH) and recombinant inbred lines (RILs) populations, derived from the same indica-japonica cross ‘Samgang × Nagdong’, were constructed to analyze the quantitative trait loci (QTLs) affecting agronomic traits in rice. The segregations of agronomic traits in RILs population showed larger variations than those in DH population. A total of 10 and 12 QTLs were identified on six chromosomes using DH population and seven chromosomes using RILs population, respectively. Three stable QTLs including pl9.1, ph1.1, and gwp11.1 were detected through different years. The percentages of phenotypic variation explained by individual QTLs ranged from 8 to 18% in the DH population and 9 to 33% in the RILs population. Twenty-three epistatic QTLs were identified in the DH population, while 21 epistatic QTLs were detected in the RILs population. Epistatic interactions played an important role in controlling the agronomic traits genetically. Four significant main-effect QTLs were involved in the digenic interactions. Significant interactions between QTLs and environments (QE) were identified in two populations. The QTLs affecting grain weight per panicle (GWP) were more sensitive to the environmental changes. The comparison and QTLs analysis between two populations across different years should help rice breeders to comprehend the genetic mechanisms of quantitative traits and improve breeding programs in marker-assisted selection (MAS).     

Genetic analyses of agronomic and seed quality traits of doubled haploid population in Brassica napus through microspore culture

Summary The results showed that the F1 genotype from the cross (Brassica napus cv. Zheshuang 758 × cv. Z-4115) had good response to embryogenesis, and their embryo yield and rate of plant regeneration reached 69.8 embryo/bud and 46.9%, respectively. Characters from the doubled haploid (DH) populations in B. napus were analyzed and it was showed that the means of each agronomic trait were between their parents, but they were nearer to the paternal in 6 agronomic traits (plant height, branch position, number of pods in the main raceme, length of pod, number of pods/plant and number of seeds/pod). The number of genes controlling each agronomic trait was analyzed based on the DH populations. The results showed that the number of genes controlling number of pods in the main raceme was the highest (15.6), and the least number of genes was involved for stem width (only 7.9). According to estimated coefficients of skewness and kurtosis of the traits tested, gene interaction was found to be absent for stem width, plant height, length of main raceme, number of primary and secondary branches, pod density in the main raceme and seed weight/plant. Complementary interaction was also observed in five agronomic traits (number of pods in the main raceme, length of pod, number of pods/plant, number of seeds/pod and 1000-seed weight). A significantly positive correlation was observed between seed yield/plant and four agronomic traits (length of main raceme, length of pod, number of pods/plant and 1000-seed weight). The experiment also showed that the erucic acid, glucosinolate, oil and protein contents of DH populations were 34.23%, 87.09 μmol/g, 44.09% and 42.67%, respectively. The numbers of genes controlling each quality trait were 7.8, 9.7, 9.4 and 8.7, respectively. Partial correlations between the seed quality traits and the agronomic characters of DH populations were analyzed. In this experiment, the partial correlations among seed quality traits were also analyzed and it was found that the oil content had a negative correlation with the other three seed quality traits.     

Field Performance of Androgenetic Doubled Haploid Spring Wheat Lines in Comparison with Lines Selected by the Pedigree System

A total of 316 doubled haploid lines (DH) of spring wheat were compared with 621 lines selected in a pedigree system (PS) under field conditions in a breeding nursery. The lines originated from 21 crosses and the samples tested represent mean values for variables comprising the main breeding goals such as disease resistance, baking quality, and agronomic traits. In general, the DH lines were later in days to heading and shorter than PS lines of different homozygosity levels. Except for leaf rust (Puccinia recondite), the DHs were more resistant to the artificially infected diseases such as powdery mildew (Erysiphe graminis), stripe rust (Puccinia striiformis), and septoria nodorum blotch (Septoria nodorum). These differences, which were relatively small for practical selection purposes, were probably due to easier and more precise disease assessment of the homozygous DH lines. For quality characters, the DHs had a higher protein level, while the rest of the parameters were similar for both origins. The data analyzed suggest so far that androgentic doubled haploids in spring wheat are very similar to lines selected in a pedigree system in respect to all the agronomic characters tested. However, the DH lines were produced in a much shorter period of time. It is suggested that androgenetic doubled haploids be produced from F₁ hybrids and that the well-established bulk method should continue to allow selection bulk method should continue to allow selection for rare recombinants as soon as homozygosity is reached.     

Field evaluation of transgenic Brassica napus lines carrying a seed-specific chimeric 2S albumin gene

The agronomic performance of transgenic Brassica napus lines carrying chimeric 2S albumin seed protein genes was studied under field conditions. A total of 37 transgenic lines were compared to non-transformed lines. Plant development, morphological characteristics, yield parameters and seed composition were considered. Only a few transgenic lines differed from the control, and then only for particular parameters: four lines had altered leaf morphology, two differed in germination efficiency, one differed in yield, and three differed in some parameters of seed composition. These differences were not correlated with the presence or expression of the transgene and, thus, were more probably linked to cell- or tissue-culture effects, or to residual variation present in cv. ‘Drakkar’. A statistical interaction between variations in a limited number of seed composition parameters and cell haploid culture was observed. While the expression of the chimeric 2S albumin gene was not as high as hoped, this study demonstrates that transgenic lines can be isolated in which the intrinsic morphological and agronomic properties of the original lines are maintained.     

The influence of photoperiod on the Vrn-H2 locus (4H) which is a major determinant of plant development and reproductive fitness traits in a facultative × winter barley (Hordeum vulgare L.) mapping population

To determine the effect of Vrn‐H2 locus on plant developmental and agronomic traits, detailed controlled environment tests involving a factorial set of vernalization and photoperiod treatments were carried out using doubled haploid lines developed from a facultative (Vrn‐H2^?) × winter (Vrn‐H2⁺) barley cross. The allele phase at the Vrn‐H2 locus influenced heading date as well as the developmental and agronomic traits. The performance of Vrn‐H2⁺ lines was significantly influenced by vernalization: reproductive fitness traits showed significant decreases without vernalization. However, the effects of alleles at the Vrn‐H2 locus extended beyond simple satisfaction of the vernalization requirement. Vrn‐H2⁺ lines showed increased reproductive fitness compared with Vrn‐H2^? lines when vernalization was followed by a long photoperiod. The responses of the two Vrn‐H2 allele classes to photoperiod duration were quite different in terms of heading date, developmental and agronomic traits. These results suggest that alleles at the Vrn‐H2 locus – and/or tightly linked gene(s) – respond primarily to the exogenous signal of vernalization (temperature), but when the vernalization requirement has been fulfilled, they also respond to photoperiod duration.     

Identification and molecular mapping of a dwarfing gene in barley (<Emphasis Type="Italic">Hordeum vulgare</Emphasis> L.) and its correlation with other agronomic traits

Ninety-two doubled haploid (DH) lines, generated from a cross between Franklin and TX9425 (a Chinese Landrace), were grown in three environments to identify quantitative trait loci (QTLs) controlling agronomic traits including heading date, plant height and spike characteristics. The DH lines showed a wide range of variations for all the agronomic traits tested. Most of the traits were controlled by one or two major QTLs which explained 9.5–80.9% of the phenotypic variation. Two dwarfing genes were identified from the cross. One of the dwarfing genes was from Franklin, which is the same as the previously reported denso gene. The other dwarfing gene was from the Chinese landrace variety. Both dwarfing genes were temperature and/or day length sensitive. The dwarfing gene from Franklin was more effective in early sowing trials (shorter day length and lower temperature) while the gene from TX9425 was more effective in later sown trials. The dwarfing gene from TX9425 was located at a similar position to the uzu gene. However, it differed from this gene being temperature sensitive with very close links to short spikes, awns and high grain density which is more like a brh gene. To effectively use this gene in a breeding program, it is necessary to break the linkage between the dwarfing gene and the unfavourable spike traits.     

Comparison of Spring Barley Lines Produced by Single Seed Descent, Pedigree Inbreeding and Doubled Haploidy

Samples of inbred lines of Hordeum vulgare produced by pedigree inbreeding, doubled haploidy and single seed descent have been compared for a range of agronomic characters. Differences between the means of the in bred s produced by single seed descent and pedigree breeding were due to differing levels of inbreeding. The present study confirmed chat recombinant inbred families which out-perform the higher scoring parent may be produced by either method. Caution should, however, be exercised when simply inherited characters controlled by major genes are fixed in early generations. Differences between the doubled haploid and single seed descent samples for the character thousand grain weight were attributable to coupling linkages involving epistatic genes. The production and evaluation of random inbred lines should be considered as an alternative to pedigree methods of barley breeding.     

Haploid induction from F1 hybrids between CMS shallot with Allium galanthum cytoplasm and common onion by unpollinated flower culture

The induction of haploid plants from F₁ hybrids between CMS shallot with Allium galanthum cytoplasm and common onion was examined. Starting with 535 unpollinated flowers cultured in B5 medium 25 seedlings from part henogenetic embryos were obtained of which 13 seedlings survived. Eleven seedlings were determined as haploid plants (2n = x = 8) and 2 seedlings were doubled haploid plants (2n = 2x = 16). All haploid and doubled haploid plants preserved chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from A. galanthum. Segregation in different characters was observed among the haploid plants. The haploid and doubled haploid plants exhibited the different combinations of genes from shallot and common onion. Crossing of the doubled haploid plants with other shallot strains, common onion cultivars or related species may produce excellent F₁ hybrids for bulb production. This revised version was published online in July 2006 with corrections to the Cover Date.     

玉米单倍体加倍(DH)系籽粒性状表现的研究

玉米单倍体育种是快速选育纯合自交系的方法.研究利用单倍体诱导系,对来自BSSS杂种优势群的两个自交系BX08和BX19的杂交F1进行诱导获得单倍体,又经加倍获得33个加倍纯合系(DH),分析单倍体加倍纯合系与两个亲本及F1杂交种的果穗及籽粒性状表现.结果表明,单倍体加倍系的籽粒容积和质量比亲本籽粒都有所提高,子代的农艺性状获得显著改善,说明发生单倍体不是基因随机分离的结果,单倍体育种存在选择压力,有利于优良基因的累积,淘汰不良基因.     

Mapping of Re,a gene conferring the red leaf trait in ornamental kale (Brassica oleracea L. var. acephala)

Variegated leaf colour is an important agronomic trait that affects the market value of ornamental kale (Brassica oleracea L. var. acephala). The red leaf phenotype in kale is due to anthocyanin accumulation. To investigate the pattern of inheritance of this trait, we constructed an F₂ population by crossing ‘Y005‐15’, a double haploid with red leaves, with a white‐leaved double haploid, ‘Y011‐13‐38’, followed by self‐pollination. An F₂ population consisting of 4284 individuals was used to study the inheritance of this trait, which showed that the character was controlled by a dominate gene. All of the 1050 white leaf trait plants in the F₂ were used for mapping and developing markers linked to Re gene. Results showed that Re was mapped to a locus on linkage group C09 of Brassica oleracea, and the locus was mapped between six SSR markers (C9Z1, C9Z16‐1, C9Z90, C9Z94, C9Z96 and C9Z99), with a genetic distance of 6.7, 1.0, 0.3, 2.0, 2.1 and 0.4 cM from Re gene, respectively. These results may facilitate marker‐assisted selection of the red leaf trait in kale breeding as well as map‐based cloning of the red leaf trait gene.     

Effects of loci determining photoperiod sensitivity (Ppd-H1) and vernalization response (Sh2) on agronomic traits in the 'Dicktoo'×'Morex' barley mapping population

The objectives of this research were to determine the individual and interaction effects of the Ppd-H1 and Sh2 loci on agronomic traits under short- and long-photoperiod regimes. Nineteen doubled haploid (DH) lines from the ‘Dicktoo’בMorex’ mapping population, which represented the four genotypes at the Ppd-H1 and Sh2 loci, were pheno-typed in controlled environment photoperiods. Both Ppd-H1 and Sh2 had significant effects on several agronomic traits, in addition to their role in determining first node appearance and flowering time. The magnitude of these effects depended on daylight. Under long-day conditions (18 h) Ppd-H1, and under short-day conditions (12 h) Sh2 was a significant determinant of most characters. The interactions between these two loci were significant for several characters, particularly for yield components, under both long- and short-photoperiod regimes. Under the long-day treatment, Ppd-H1 influenced plant height through the determination of node number. There was an epistatic association between the two loci for both 1000-kernel weight and tillering. The combination of photoperiod insensitivity and vernalization requirement caused a significant increase in tillering. This was paralleled by a decrease in 1000-kernel weight. Under the long-day treatment, neither Ppd-H1 nor Sh2 influenced plant yield. Under short-day conditions, the combination of photoperiod insensitivity and vernalization requirement had a pronounced negative effect on plant yield.     

Potential use of doubled haploid lines for the screening of resistance to yellow rust (Puccinia striiformis) in hexaploid wheat

Doubled haploid lines derived from anther culture of two Iranian spring wheat genotypes‘Ghods’susceptible and‘9106’resistant to yellow rust in Iranian field conditions, and their F₁ hybrids were used in this study. Seedlings of 36 doubled haploid lines, selected out of 96 according to their agronomic traits and the two parental genotypes were inoculated with eight races of yellow rust. The parental genotypes (‘Ghods’and‘9106’) were segregating for some of the races but their doubled haploid lines were either resistant or susceptible to them.‘Ghods’was susceptible to three of the races studied but three doubled haploid lines derived from it were resistant to them. Five selected doubled haploids from the‘9106’genotype and six from F₁ hybrid plants were resistant to all eight races tested. After further investigations in Iranian field conditions it was found that some of these lines can be used as donor genotypes for resistance to yellow rust in wheat breeding programmes. Use of these genotypes should be possible if the French yellow rust races used for selection also represent the dominant races in Iran. It can be concluded that anther culture provides an efficient method for fixing genes of resistance to yellow rust and desirable doubled haploids from F₁ plants can be derived.     

The evaluation of superior Hordeum bulbosum L. genotypes for use in a doubled haploid barley breeding programme

Summary Eight Hordeum bulbosum selections were produced from a cross between Cb 2920/4 and Cb 2929/1, two genotypes widely used in doubled haploid breeding programmes. The selections were hybridized with barley to evaluate their ability to produce high proportions of well-differentiated haploid embryos compared with Cb 2929/1 as control. We report here an initial small-scale investigation followed by a larger-scale test in two different environments to assess seed setting, haploid embryo differentiation rates and VB hybrid formation. These VB embryos contain both parental sets of chromosomes and occur more frequently in the glasshouse during the winter. Two of the eight selections were identified as combining the desirable characteristics of both parents, namely high seed setting on cv. Vada which is partially incompatible with H. bulbosum, large numbers of well-differentiated haploid embryos and a low incidence of VB hybrids. The selections are available for release to interested research workers and plant breedersAbbreviations DH doubled haploid - VB a hybrid from H. vulgare x H. bulbosum which contains both parental sets of chromosomes