Confirmation of intraspecific crossing and single and joint segregation of biochemical loci ofVolvariella volvacea

Nineteen lines ofV. volvacea were obtained from worldwide sources and analyzed for allozyme activity. Twelve loci (Aat, Acp-2, Dia, Est-4, Gipi, Lap-1, Pep-GL, Pep-LLL-1, Pep-LLL-2, Pgm, Pgd, andSod) were monomorphic; 5 loci (Ada, Gpt, Mpi, Np, andPep-PAP) were polymorphic. Nine unique genotypes were distinguished among the 19 lines. Multilocus enzyme electrophoresis was used to confirm intraspecific crosses between putative homokaryons. Putative homokaryons were recognized by electrophoresis of single-spore-derived cultures and used as breeding stock. These breeding stocks were grown together in dual culture on agar, and selections were made from the interaction zone. These selections were then transferred to make spawn. Intraspecific crosses were confirmed by the presence of heteromeric allozymes. To determine single and joint segregation of biochemical loci, 84 single-spore-derived offspring from a confirmed hybrid sporocarp with five heterozygous loci were analyzed. Mendelian segregation was verified for the five loci. One linkage (Np withGpt,r=0.0) was found among 10 pairwise comparisons for joint segregation. The basidiospores ofV. volvacea are haploid and homokaryotic isolates may mate to yield heterokaryons.     

Genetic Variation and Combining Ability Analysis of Bruising Sensitivity in Agaricus bisporus

Advanced button mushroom cultivars that are less sensitive to mechanical bruising are required by the mushroom industry, where automated harvesting still cannot be used for the fresh mushroom market. The genetic variation in bruising sensitivity (BS) of Agaricus bisporus was studied through an incomplete set of diallel crosses to get insight in the heritability of BS and the combining ability of the parental lines used and, in this way, to estimate their breeding value. To this end nineteen homokaryotic lines recovered from wild strains and cultivars were inter-crossed in a diallel scheme. Fifty-one successful hybrids were grown under controlled conditions, and the BS of these hybrids was assessed. BS was shown to be a trait with a very high heritability. The results also showed that brown hybrids were generally less sensitive to bruising than white hybrids. The diallel scheme allowed to estimate the general combining ability (GCA) for each homokaryotic parental line and to estimate the specific combining ability (SCA) of each hybrid. The line with the lowest GCA is seen as the most attractive donor for improving resistance to bruising. The line gave rise to hybrids sensitive to bruising having the highest GCA value. The highest negative SCA possibly indicates heterosis effects for resistance to bruising. This study provides a foundation for estimating breeding value of parental lines to further study the genetic factors underlying bruising sensitivity and other quality-related traits, and to select potential parental lines for further heterosis breeding. The approach of studying combining ability in a diallel scheme was used for the first time in button mushroom breeding.     

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     

Sexuality and Genetic Identity in the Agaricus Section Arvenses

Twelve wild collections and one commercial strain were used to characterize breeding systems and to develop molecular identities in the Arvenses section of the genus Agaricus, which includes the “horse mushroom” A. arvensis. Two morphotypes were identified based on macro- and micromorphological features. However, not all collections could be delimited by conventional taxonomic characters. Sequencing of the small subunit intergenic spacer (ITS) region (368 to 370 bp) of the rRNA genes clearly resolved the 13 collections into two clusters consistent with the identified morphotypes. Single-spore progenies and mating type testers were established and used to test intra- and interstock compatibility. The two compatibility groups identified were consistent with ITS clusters. Compatibility group I stocks readily interbred within the constraints of a unifactorial heterothallic system with a multiallelic mating type factor. Compatibility group II had a more restricted breeding pattern, and interactions were difficult to predict on the basis of mating type. Morphological data, ITS sequences, and the ability to interbreed suggest that these collections are part of a complex of interrelated species. Single-spore, homokaryotic isolates from both compatibility groups were able to fruit in compost culture, and two of the collections may represent natural homokaryotic fruiting. We conclude that species from the section Arvenses have versatile unifactorial heterothallic life cycles that permit both interbreeding and homokaryotic fruiting.     

Additional sources of resistance to groundnut rosette disease in groundnut germplasm and breeding lines

Groundnut rosette, a virus disease of groundnut (Arachis hypogaea) transmitted by the aphid, Aphis craccivora Koch, reduces yield in susceptible cultivars by 30–100%. Additional sources were sought in germplasm accessions involving 2301 lines from different sources and from 252 advanced breeding lines derived from crosses involving earlier identified sources of resistance to rosette. The lines were evaluated in field screening trials using an infector row technique during 1996 and 1997 growing seasons. Among the germplasm lines, 65 accessions showed high levels of resistance while 134 breeding lines were resistant. All rosette disease resistant lines were susceptible to groundnut rosette assistor virus. This work identified germplasm and breeding lines that will contribute to an integrated management of groundnut rosette disease. These new sources also provide an opportunity to eliminate yield losses due to the rosette disease.     

Amplified fragment length polymorphisms as a tool for DNA fingerprinting sunflower germplasm: genetic diversity among oilseed inbred lines

 Amplified fragment length polymorphism (AFLP) analysis is a rapid and efficient method for producing DNA fingerprints. The AFLP diversity of sunflower has not been described, and much of the public germ plasm of sunflower has not yet been fingerprinted. Our objectives were to: (1) estimate genetic similarities, polymorphism rates, and polymorphic information contents (PICs) for AFLP markers among elite public oilseed inbred lines, and (2) assess the genetic diversity of inbred lines using genetic similarities estimated from AFLP fingerprints. We produced fingerprints for 24 public inbred lines of sunflower (Helianthus annuus L.) using six AFLP primer combinations. These primers produced a total of 359 AFLP markers or about 60 markers per primer combination. Genetic similarities ranged from 0.70 to 0.91, polymorphism rates ranged from 7 to 24%, and PICs ranged from 0.0 to 0.5. Genetic similarities were lower overall for maintainer (B)×restorer (R) crosses than for B×B or R×R crosses. Principal-coordinate and cluster analyses separated lines into two groups, one for B-lines and another for R-lines. These groupings illustrate the breeding history and basic heterotic pattern (B×R) of sunflower and the widespread practice of using B×B and R×R crosses to develop new lines. There were, nevertheless, distinct subgroups within these groups. These subgroups may represent unique heterotic groups and create a basis for formally describing heterotic patterns in sunflower. Received: 10 June 1996 / Accepted: 4 April 1997     

Extranuclear chloramphenicol resistance mutations in the basidiomyceteSistotrema brinkmannii

Four chloramphenicol resistance mutations were induced in homokaryons ofSistotrema brinkmannii and a heterokaryon test was used to distinguish whether the mutations were nuclear or extranuclear. Dikaryons were recovered from sexually compatible pairings of chloramphenicol-resistant and sensitive strains which also carried chromosomal markers (mating-type and auxotrophic). The homokaryotic components of the dikaryons were recovered from vegetative cells by the production and regeneration of protoplasts. The homokaryons derived from protoplasts were tested for chloramphenicol resistance/sensitivity as well as for the chromosomal markers. In heterokaryon tests, the chloramphenicol resistance/sensitivity determinants reassociated with the chromosomal markers. Only a single recombinant with respect to chromosomal markers was observed among a total of 220 homokaryons analyzed. All pairs of the same chromosomal markers produced numerous recombinants in sexual crosses. These results indicated that the chloramphenicol resistance mutations were extranuclear.     

Amplification of the high production and low production nature of antibodies produced by the genetic selection of mice. Correlation between immunodeficiency and the incidence of lymphoma

Five bidirectional selective breedings have been carried out in mice for quantitative antibody responses to various natural immunogens. Appropriate crosses between the five "high responder" lines and between the five "low responder" lines resulting from these selections, were performed to obtain balanced hybrid populations. These populations were further selected for antibody production on the basis of multiple responses i.e., responses to all the antigens used in the previous separate selections. The result was a marked amplification of the interline divergence. The low line in particular has a profound deficiency for both primary and secondary responses. Moreover a high incidence of lymphomas was observed among these low responder mice (up to 42%).     

Effects of Nematicides and Plant Resistance on White Clover Performance and Seasonal Populations of Nematodes Parasitizing White Clover in Grazed Pasture

Root-infecting nematodes are a major cause of white clover, Trifolium repens, not reaching its potential in New Zealand pastures. Resistance and/or tolerance are the preferred control options. Greenhouse-based, recurrent selection programs have developed resistance to Meloidogyne trifoliophila and Heterodera trifolii, and a field-based program has developed tolerance. Lines from these programs were compared with commercial cultivars as controls in a series of field trials at four sites over 4 years. Resistant lines from the CCN program performed better than susceptible lines and as well as most cultivars, reflecting the high level of resistance developed in this greenhouse-based program. In stained root from Cambridge, numbers of CCN were lower in resistant lines than in cultivars; numbers in susceptible lines were intermediate. CCN resistance was also reflected to a lesser extent in the number of cysts counted in soil under resistant lines in Palmerston North. The root-knot nematode-resistant material performed better than the susceptible and as well as most cultivars. In one trial of CRKN-resistant lines, resistant and susceptible lines had similar numbers of CRKN which were both lower than the numbers in the cultivars; in the second trial, there were fewer CRKN in resistant than in susceptible lines or cultivars. The tolerant selections, developed under field conditions, performed as well as or better than the cultivars. The selections from the breeding programmes have exhibited strong agronomic potential across locations and years, and the best material has been crossed; progeny are being assessed in current field trials.     

Genomic Predictability of Interconnected Biparental Maize Populations

Intense structuring of plant breeding populations challenges the design of the training set (TS) in genomic selection (GS). An important open question is how the TS should be constructed from multiple related or unrelated small biparental families to predict progeny from individual crosses. Here, we used a set of five interconnected maize (Zea mays L.) populations of doubled-haploid (DH) lines derived from four parents to systematically investigate how the composition of the TS affects the prediction accuracy for lines from individual crosses. A total of 635 DH lines genotyped with 16,741 polymorphic SNPs were evaluated for five traits including Gibberella ear rot severity and three kernel yield component traits. The populations showed a genomic similarity pattern, which reflects the crossing scheme with a clear separation of full sibs, half sibs, and unrelated groups. Prediction accuracies within full-sib families of DH lines followed closely theoretical expectations, accounting for the influence of sample size and heritability of the trait. Prediction accuracies declined by 42% if full-sib DH lines were replaced by half-sib DH lines, but statistically significantly better results could be achieved if half-sib DH lines were available from both instead of only one parent of the validation population. Once both parents of the validation population were represented in the TS, including more crosses with a constant TS size did not increase accuracies. Unrelated crosses showing opposite linkage phases with the validation population resulted in negative or reduced prediction accuracies, if used alone or in combination with related families, respectively. We suggest identifying and excluding such crosses from the TS. Moreover, the observed variability among populations and traits suggests that these uncertainties must be taken into account in models optimizing the allocation of resources in GS.     

Genetic recombination in sexual crosses of phycomyces

Sexual crosses between strains of Phycomyces blakesleeanus , involving three auxotrophic and one color marker and yielding a high proportion of zygospore germination, are described. Samples of 20–40 germ spores from 311 individual fertile germ sporangia originating from five two-factor and three three-factor crosses were characterized. The results show: (1) absence of any contribution of apogamic nuclei to the progeny, (2) confirmation of Burgeff's conjecture that the germ spores of any germ sporangium in most cases derive from one meiosis. In a cross involving two allelic markers the analysis of 175 pooled germ sporangia suggests an intragenic recombination frequency of 0.6%. All other factor combinations tested are unlinked. The bulk of the germ spores are homokaryotic. However, a small portion (4%) are heterokaryotic with respect to mating type.     

Identification,introgression, and molecular marker genetic analysis and selection of a highly effective novel oat crown rust resistance from diploid oat, <Emphasis Type="Italic">Avena strigosa</Emphasis>

Key message

Oat crown rust is one of the most damaging diseases of oat. We identified a new source of resistance and developed KASP and TaqMan markers for selection in breeding programs.

Abstract

A new highly effective resistance to oat crown rust (Puccinia coronata f. sp. avenae) was identified in the diploid oat Avena strigosa PI 258731 and introgressed into hexaploid cultivated oat. Young plants with this resistance show moderate susceptibility, whereas older plant tissues and adult plants are resistant with no virulent isolates encountered in over 8 years of testing. Resistance was incorporated into hexaploid oat by embryo rescue, colchicine chromosome doubling followed by backcrosses with a hexaploid parent, and selection for stable transmission of resistance. To mitigate flag leaf and panicle chlorosis/necrosis associated with the resistance, crosses were made with derived resistant lines to breeding lines of divergent parentage followed by selection. Subsequently, two F₂ sister lines, termed MNBT1020-1 and MNBT1021-1, were identified in which the chlorosis/necrosis was reduced. These two lines performed well in replicated multi-location state trials in 2015 and 2016 out-yielding all cultivar entries. Segregating F_2:3 plants resulting from crosses of MNBT lines to susceptible parents were genotyped with the oat 6K SNP array, and SNP loci with close linkage to the resistance were identified. KASP assays generated from linked SNPs showed accurate discrimination of the resistance in derivatives of the resistant MNBT lines crossed to susceptible breeding lines. A TaqMan marker was developed and correctly identified homozygous resistance in over 95% of 379 F₄ plants when rust was scored in F_4:5 plants in the field. Thus, a novel highly effective resistance and associated molecular markers are available for use in breeding, genetic analysis, and functional studies.

     

Exploitation of the resistance to carrot fly in the wild carrot species Daucus capillifolius

Under field conditions a wild Daucus species from Libya, D. capillifolius, supported less than one tenth as many carrot flies (Psila rosae) as the susceptible carrot cultivar Danvers Half Long 126. Breeding lines developed from crosses between D. capillifolius and three different carrot types were grown in a series of field experiments at Wellesbourne between 1980 and 1989. Each year selections were made for agronomic quality and/or for increased resistance to carrot fly. The programme produced lines which for size, shape and colour represented most of the commercially-important carrot types. Some of these lines were also significantly more resistant to carrot fly than selections from the partially-resistant cv. Sytan. However, the best lines were not as resistant as the wild parent. The highest quality resistant lines were sold to seed companies for variety production.     

Genetic control of immunity to turnip mosaic virus in winter oilseed rape (Brassica napus ssp. oleifera) and the effect of foreign isolates of the virus

Immunity to a UK isolate (UK 1) of turnip mosaic virus (TuMV) was studied in eight lines of oilseed rape selected from cv Rafal. Six of these lines were uniformly immune and two segregated. Segregation ratios in the F₂ generation of reciprocal crosses between two uniformly immune lines and two uniformly susceptible cultivars (Mikado and Yeoman) showed that immunity was controlled by a dominant nuclear allele. The immunity was confirmed by the inability to detect virus particles in mechanically inoculated plants by back inoculations, ELISA and ISEM tests. Plants were immune to repeated inoculations and aphid transmissions. The immunity was effective against one other UK isolate and two German isolates of TuMV. Another UK isolate (UK 3) and a Greek isolate partially overcame the immunity causing local infection and a Canadian and a Danish isolate overcame it completely causing systemic mosaic-type symptoms. When these immunity-breaking strains were tested against swede line 165 which is also immune to UK 1 TuMV, the Canadian isolate overcame the immunity whereas the Danish isolate did not. Using this swede line, one susceptible and one immune line of oilseed rape as differentials, four distinct groups of TuMV isolates could be identified. Selections of oilseed rape immune to UK TuMV isolates were more severely affected by the Canadian TuMV than UK TuMV susceptible selections. The gene determining immunity to TuMV had no pleiotropic effect on susceptibility to cauliflower mosaic virus. The implications on these findings in relation to breeding for virus resistance are discussed.     

Evidence for Outcrossing via the Buller Phenomenon in a Substrate Simultaneously Inoculated with Spores and Mycelium of Agaricus bisporus

In Agaricus bisporus, traditional cultivars and most of the wild populations belong to A. bisporus var. bisporus, which has a predominantly pseudohomothallic life cycle in which most meiospores are heterokaryons (n + n). A lower proportion of homokaryotic (n) meiospores, which typify the heterothallic life cycle, also are produced. In wild populations, pseudohomothallism was thought previously to play a major role, but recent analyses have found that significant outcrossing also may occur. We inoculated a standard substrate for A. bisporus cultivation simultaneously with homokaryotic mycelium from one parent and spores from a second parent. Culture trays produced numerous sporocarps that could theoretically have resulted from five different reproductive modes (pseudohomothallism, selfing or outcrossing via heterothallism, and selfing or outcrossing via the Buller phenomenon [i.e., between a homokaryon and a heterokaryon]). Most or all of the sporocarps resulted from outcrossing between the inoculated homokaryon and the inoculated heterokaryotic spores (or mycelia that grew from them). These data broaden our understanding of population dynamics under field conditions and provide an outcrossing method that could be used in commercial breeding programs.     

Basal immunoglobulin serum concentration and isotype distribution in relation to the polygenic control of antibody responsiveness in mice

Serum Ig concentration and isotype distribution were determined in the high (H) and low (L) responder lines selected for antibody response to complex immunogens. Data were recorded in normal and postimmunization sera from the H and L lines produced by five independent selective breedings (selections I, 11, III, IV, and V). Ig levels were much higher in H than in L mice of all the selections. In four selections this interline difference increased further after immunization with the selection antigens. This is in agreement with the general effect of the polygenic control of antibody responses operating in H and L lines. The Ig isotype profiles of normal sera were different in each line; however, similitudes were noticed between H and L lines in selections I and If. In contrast, in selections III, IV, and V a similar interline difference was observed: the lack of IgG2a isotype in L lines only. After immunization there were minor alterations of the isotype profiles except in the H lines of selections III and IV, in which a clear inverse modification of IgG₁ and IgG_2a proportions occurred. The characteristic pattern of each selection may be partially dependent on isotype-restricted regulatory effects in relation to the immunization procedure used for selective breeding.     

Linkage analysis of quantitative trait loci in multiple line crosses

Simple line crosses, for example, backcross and F₂, are commonly used in mapping quantitative trait loci (QTL). However, these simple crosses are rarely used alone in commercial plant breeding; rather, crosses involving multiple inbred lines or several simple crosses but connected by shared inbred lines may be common in plant breeding. Mapping QTL using crosses of multiple lines is more relevant to plant breeding. Unfortunately, current statistical methods and computer programs of QTL mapping are all designed for simple line crosses or multiple line crosses but under a regular mating system. It is not straightforward to extend the existing methods to handle multiple line crosses under irregular and complicated mating designs. The major hurdle comes from irregular inbreeding, multiple generations, and multiple alleles. In this study, we develop a Bayesian method implemented via the Markov chain Monte Carlo (MCMC) algorithm for mapping QTL using complicated multiple line crosses. With the MCMC algorithm, we are able to draw a complete path of the gene flow from founder alleles to their descendents via a recursive process. This has greatly simplified the problem caused by irregular mating and inbreeding in the mapping population. Adopting the reversible jump MCMC algorithm, we are able to simultaneously search for multiple QTL along the genome. We can even infer the posterior distribution of the number of QTL, one of the most important parameters in QTL study. Application of the new MCMC based QTL mapping procedure is demonstrated using two different mating designs. Design I involves two inbred lines and their derived F₁, F₂, and BC populations. Design II is a half-diallel cross involving three inbred lines. The two designs appear different, but can be handled with the same robust computer program.     

The strategy and potential utilization of temperate germplasm for tropical germplasm improvement: a case study of maize (<Emphasis Type="Italic">Zea mays</Emphasis> L.)

The organization of maize (Zea mays L.) germplasm into genetically divergent heterotic groups is the foundation of a successful hybrid maize breeding program. In this study, 94 CIMMYT maize lines (CMLs) and 54 United States germplasm enhancement of maize (GEM) lines were assembled and characterized using 1,266 single nucleotide polymorphisms (SNPs) with high quality. Based on principal component analysis (PCA), the GEM lines and CMLs were clearly separated. In the GEM lines, there were two groups classified by PCA corresponding to the heterotic groups “stiff stalk” and “non-stiff stalk”. CMLs did not form obvious subgroups by PCA. The allelic frequency of each SNP differed in GEM lines and CMLs. In total, 3.6% alleles (46/1,266) of CMLs are absent in GEM lines and 4.4% alleles (56/1,266) of GEM lines are absent in CMLs. The performance of F1 plants (n = 654) produced by crossing between different groups based on pedigree information was evaluated at the breeding nurseries of two CIMMYT stations. Genomic estimated phenotypic values of plant height and days to anthesis for a testing set of 45 F1 crosses were predicted based on the training data of 600 F1 crosses using a best linear unbiased prediction method. The prediction accuracy benefitted from the adoption of the markers associated with quantitative trait loci for both traits; however, it does not necessarily increase with an increase in marker density. It is suggested that genomic selection combined with association analysis could improve prediction efficiency and reduce cost. For hybrid maize breeding in the tropics, incorporating GEM lines which have unique alleles and clear heterotic patterns into tropically adapted lines could be beneficial for enhancing heterosis in grain yields.     

A comparison of anther and microspore culture as a breeding tool in Brassica napus

Summary A direct comparison of microspore culture and anther culture was made in Brassica napus using F₁ crosses of Regent (canola) by Golden (rapeseed), and their reciprocals, as well as a hybrid between Reston and a highly embryogenic, canola-quality breeding line (G231) as donor plants. The study confirmed that microspore culture can be ten times more efficient than anther culture for embryo production. Embryo yields from cultures initiated from the Reston x G231 were four-fold greater than those initiated from the Regent x Golden crosses, and significant differences were also detected among cultures initiated from the different Regent x Golden crosses. These results illustrate the influence that donor plant genotype has on embryo production. However, superior embryogenic potential among donor material was not always coincident with superior plant production. The average haploid-todiploid ratio in microspore-derived regenerates was 21 for the population obtained from the Regent x Golden crosses but 11 for the Reston x G231 cross. For both types of material, the frequency of diploids increased upon repeated cycles of explanting. A field study showed that there were no differences between the populations of anther-derived and microspore-derived spontaneous diploid and doubled haploid lines, with respect to the days required for them to flower or to mature. The information is valuable for canola breeding programs considering the use of haploidy.