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.     

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.     

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     

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.     

Genetical investigations into β-glucan content in barley

Summary Random inbred lines produced by doubled haploidy (DH) and single seed descent (SSD) have been used to investigate the genetics of -glucan (gum) content in barley (Hordeum vulgare). Genetical analyses indicated that gum content is controlled by a simple additive genetic system. Significant negative genetic correlations were observed between -glucan content, thousand grain weight and height in the DH samples. These correlations were much reduced in the SSD samples and would suggest linkage of the genes controlling these characters. The presence of repulsion linkages could be exploited in a barley breeding programme by producing F1 derived DH to generate recombinants with high thousand grain weight and low -glucan content. Genetical parameters estimated from DH and F3 samples have successfully been used to predict the number of inbred lines transgressing the parental range for -glucan content and bivariate combinations involving -glucan.     

Quantitative trait analysis of seed yield and other complex traits in hybrid spring rapeseed (Brassica napus L.): 1. Identification of genomic regions from winter germplasm

The introgression of winter germplasm into spring canola (Brassica napus L.) represents a novel approach to improve seed yield of hybrid spring canola. In this study, quantitative trait loci (QTL) for seed yield and other traits were genetically mapped to determine the effects of genomic regions introgressed from winter germplasm into spring canola. Plant materials used comprised of two populations of doubled haploid (DH) lines having winter germplasm introgression from two related French winter cultivars and their testcrosses with a spring line used in commercial hybrids. These populations were evaluated for 2 years at two locations (Wisconsin, USA and Saskatchewan, Canada). Genetic linkage maps based on RFLP loci were constructed for each DH population. Six QTL were detected in the testcross populations for which the winter alleles increased seed yield. One of these QTL explained 11 and 19% of the phenotypic variation in the two Canadian environments. The winter allele for another QTL that increased seed yield was linked in coupling to a QTL allele for high glucosinolate content, suggesting that the transition of rapeseed into canola could have resulted in the loss of favorable seed yield alleles. Most QTL for which the introgressed allele decreased seed yield of hybrids mapped to genomic regions having homoeologous non-reciprocal transpositions. This suggests that allelic configurations created by these rearrangements might make an important contribution to genetic variation for complex traits in oilseed B. napus and could account for a portion of the heterotic effects in hybrids. Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Quantitative trait analysis of seed yield and other complex traits in hybrid spring rapeseed (Brassica napus L.): 2. Identification of alleles from unadapted germplasm

Unadapted germplasm may contain alleles that could improve hybrid cultivars of spring oilseed Brassica napus. Quantitative trait loci (QTL) mapping was used to identify potentially useful alleles from two unadapted germplasm sources, a Chinese winter cultivar and a re-synthesized B. napus, that increase seed yield when introgressed into a B. napus spring hybrid combination. Two populations of 160 doubled haploid (DH) lines were created from crosses between the unadapted germplasm source and a genetically engineered male-fertility restorer line (P1804). A genetically engineered male-sterile tester line was used to create hybrids with each DH line (testcrosses). The two DH line populations were evaluated in two environments and the two testcross populations were evaluated in three or four environments for seed yield and other agronomic traits. Several genomic regions were found in the two testcross populations which contained QTL for seed yield. The map positions of QTL for days to flowering and resistance to a bacterial leaf blight disease coincided with QTL for seed yield and other agronomic traits, suggesting the occurrence of pleiotropic or linked effects. For two hybrid seed yield QTL, the favorable alleles increasing seed yield originated from the unadapted parents, and one of these QTL was detected in multiple environments and in both populations. In this QTL region, a chromosome rearrangement was identified in P1804, which may have affected seed yield.Electronic Supplementary Material Supplementary material is available to authorised users in the online version of this article at .     

Development of a novel recessive genetic male sterility system for hybrid seed production in maize and other cross‐pollinating crops

We have developed a novel hybridization platform that utilizes nuclear male sterility to produce hybrids in maize and other cross‐pollinating crops. A key component of this platform is a process termed Seed Production Technology (SPT). This process incorporates a transgenic SPT maintainer line capable of propagating nontransgenic nuclear male‐sterile lines for use as female parents in hybrid production. The maize SPT maintainer line is a homozygous recessive male sterile transformed with a SPT construct containing (i) a complementary wild‐type male fertility gene to restore fertility, (ii) an α‐amylase gene to disrupt pollination and (iii) a seed colour marker gene. The sporophytic wild‐type allele complements the recessive mutation, enabling the development of pollen grains, all of which carry the recessive allele but with only half carrying the SPT transgenes. Pollen grains with the SPT transgenes exhibit starch depletion resulting from expression of α‐amylase and are unable to germinate. Pollen grains that do not carry the SPT transgenes are nontransgenic and are able to fertilize homozygous mutant plants, resulting in nontransgenic male‐sterile progeny for use as female parents. Because transgenic SPT maintainer seeds express a red fluorescent protein, they can be detected and efficiently separated from seeds that do not contain the SPT transgenes by mechanical colour sorting. The SPT process has the potential to replace current approaches to pollen control in commercial maize hybrid seed production. It also has important applications for other cross‐pollinating crops where it can unlock the potential for greater hybrid productivity through expanding the parental germplasm pool.     

Isolated microspore culture of oat (Avena sativa L.) for the production of doubled haploids: effect of pre-culture and post-culture conditions

The production of doubled haploid (DH) plants from microspores is an important technique used in plant breeding programs and basic research. Although doubled haploidy efficiencies in wheat and barley are sufficient for breeding purposes, oat (Avena sativa L.) is considered recalcitrant. The objective of this project was to develop a protocol for the production of microspore-derived embryos of oat and further develop these embryos into fertile DH plants. A number of experiments were conducted evaluating the factors influencing microspore embryogenesis, i.e. donor plant conditions, pretreatments, media composition, and culture conditions. The initial studies yielded little response, and it was not until high microspore densities (10⁶ microspores/mL and greater) were used that embryogenesis was achieved. Depending on the treatment, yields of over 5,000 embryos/10⁶ microspores were obtained for breeding line 2000QiON43. The doubled haploidy protocol includes: a 0.3 M mannitol pretreatment of the tillers for 7 days, culture in W14 basal medium with a pH of 6.5–7.5, a microspore density of 10⁶ microspores/mL, and continuous incubation at 28 °C incubation. The resulting embryos observed after 28 days were plated onto solidified W14 medium with 0.8 or 1.0 g/L activated charcoal. A colchicine treatment of 0.2 % colchicine for 4 h resulted in conversion of 80 % of the plants from haploid to DH. This protocol was successful for the production of oat microspore-derived embryos and DH green plants with minimal albinism. DH seed was produced and planted for evaluation in a field nursery.     

Comparison of single seed descent,selective intermating and mass selection for seed size in greengram (Vigna radiata (L.) Wilczek)

Summary Three selection methods (single seed descent (SSD), mass selection and selective intermating) were applied simultaneously to a highly heterogeneous and broadly based population of greengram. Progeny developing after two cycles of selection were evaluated for yield and seven other economic characters. The relative efficacy of each selection method was judged on the basis of the number of high yielding progeny, mean yield of top 10% progeny, and mean of the highest yielding progeny. Selection after two cycles of selective intermating was found to be the best method for generating productive progeny although mass selection favouring smaller seeds was an equally efficient method. Both of these were found superior to SSD selection.     

Genotypic Mapping Using RFLP Markers on Doubled Haploids Derived from a Cross Between indica and japonica Rice

A rice RFLP map with 233 loci based on a population of 81 doubled haploids (DH) from the indica/japonica hybrid of "Gui 630'/"02428" was used to analyse the genome ratios and graphical genotypes of DH lines. The “Gui 630” genome ratio of the individual DH lines varied from 29.3% to 78.6% with an average of 49% while the ratios of most DH lines ranged between 44% and 49%. Of the mapped RFLP markers 130 showed skewed segregations with a significant deviation from the expected monogenic ratio, but the numbers of the markers deviated towards male and female parent were approximately equal. It was found that the markers with the segregation deviation in the same direction tend to cluster on some chromosomes and some of their regions. The average "Gui 630' genome ratios of different chromosomes in the DH population varied greatly between 29% and 65 %. In addition, several chromosomes were inherited completely from either one of the parents in some DH lines, indicating the rare occurrence of crossover along the pairing homologous chromosomes during meiosis.     

利用RFLP标记分析一对水稻籼粳交双单倍体的基因型

对一对籼(Oryza sativa ssp.indica)粳(O.sativa ssp.japonica)(“圭630”/“02428”)杂种进行花药培养获得81个双单倍体(DH),构建了有233个RFLP标记的水稻遗传连锁图谱。对各DH系的“圭630”等位基因组频率及图示基因型进行了分析。结果表明,该DH群体的平均基因组频率为49％,各DH系分布在29.3％～78.6％之间,集中分布在44％～49％之间;有130个RFLP标记出现显著的偏分离,偏父本与偏母本分离的标记数基本相等;同向偏分离的标记常集中在一些染色体或染色体的某些区段;各染色体的平均基因组频率分布在29％～65％之间,出现明显的偏父或偏母分离;一些DH中出现其特征完全来于某一亲本的染色体,可能与这些染色体在减数分裂期的同源配对和交换有关。     

Experimental evaluation of the potential of tropical germplasm for temperate maize improvement

 Commercial maize (Zea mays L.) in the USA has a restricted genetic base as newer hybrids are largely produced from crosses among elite inbred lines representing a small sample (predominantly about 6- to 8-base inbreds) of the Stiff stalk and Lancaster genetic backgrounds. Thus, expansion of genetic diversity in maize has been a continuous challenge to breeders. Tropical germplasm has been viewed as a useable source of diversity, although the integration of tropical germplasm into existing inbred line and hybrid development is laborious. The present study is an evaluation of the potential of tropical germplasm for temperate maize improvement. All possible single-, three-way-, and double-cross hybrids among three largely temperate and three temperate-adapted, all-tropical inbred lines were evaluated in yield-trial tests. Single-cross hybrids containing as much as 50–60% tropical germplasm produced 8.0 t ha^-1 of grain yield, equivalent to the mean yield of the commercial check hybrids. On the other hand, three-way and double-cross hybrids with the highest mean yield contained lower amounts of tropical germplasm, 10–19% and 34–44%, respectively. Overall, hybrids containing 10–60% tropical germplasm yielded within the range of the commercial hybrid checks. Hybrids with more than 60% tropical germplasm had significantly lower yields, and 100% tropical hybrids yielded the least among all hybrids evaluated. The results indicate that inbred lines containing tropical germplasm are not only a useful source to expand the genetic diversity of commercial maize hybrids, but they, also are competitive in crosses with temperate materials, producing high-yielding hybrids. These experimental hybrids exhibited good standability (comparable to the commercial check hybrids) but contained 1–2% higher grain moisture, leading to delayed maturity. Recurrent selection procedures are being conducted on derivatives of these materials to extract lines with superior yield, good standability, and reduced grain moisture which can be used for commercial exploitation. Received: 26 January 1998 / Accepted: 14 July 1998     

禾木科植物染色体消除型远缘杂交的研究进展

植物远缘杂交是作物育种中广泛应用的技术。除了核型稳定的种间杂交可以获得杂种以外,还可以利用核型不稳定的种间杂交后父本染色体消除的现象,通过胚培养和染色体加倍处理获得加倍单倍体(DH)植株。然而从小麦×玉米杂交获得的DH后代与其理论上应完全同质的遗传表现却不相符,总有2～5％的DH植株发生了形态学变异。最近的研究证明。通过小麦×玉米的受精作用,一些玉米特异DNA可以被转移到小麦DH后代的基因组中。     

Gynogenetic haploid plants analysis for agronomic and enzymatic markers in maize (Zea mays L.)

Summary Experiments were conducted to investigate whether selection occurs during the processes involved in the production of doubled haploids. Haploid plants produced from two hybrids, each heterozygous for isozyme markers, were subjected to genetic analysis. The distributions of doubled haploid lines and pedigree lines derived from the hybrid C123 x Oh7 were compared with regard to agronomic character. The results suggest that the populations of haploid plants obtained by in vivo gynogenesis represent a random gametic array. Thus, in order to introduce haploid plants into breeding programmes in maize, maternal haploidy seems to be a very attractive method.     

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.