Maternal induced haploids in rye (Secale cereale L.)

Summary An inbred line of rye named P8 was found to possess haploid plants in their diploid stock. Phenomenon of spontaneous haploidy was examined in this line. Fifty families derived from this inbred line were screened to determine their ploidy level by mitotic chromosome count taking root tips. Seven families were found to contain haploid plants. Among these seven families, plants from two families which were diploid were crossed reciprocally with the standard out breeding cultivar Conrah. As all of the tested plants of P8 were found aa allele for 6-Phosphogluconic dehydrogenase (6-PGD) isozyme, therefore, only those Conrah plants which have bb alleles for the same isozyme were used in crossing programme.F1 seedlings were screened with the same isozyme marker and out of 1741 F1 seedlings tested, 51 were found to be non-hybrid phenotype for 6-PGD enzyme. Among these, only five plants were found haploid and the rest were diploid. Haploid plants were obtained only from those crosses where P8 was female which indicates that haploids in this line was maternally induced.     

Identification of seven telotrisomics of rye (Secale cereale L.)

Summary Seven monotelotrisomics emerged spontaneously from primary trisomics of rye cv. Esto. Analyses of karyotype, of chromosome pairing in meiosis and of Giemsa banding pattern were used to identify monotelotrisomics in rye as IRS, 1RL, 3RS, 4RS, 4RL, 6RS and 7RL. Vitality and fertility were better than in primary trisomics. The extra telocentrics were well transmitted through the female gamete. Transmission through the male gamete was in all cases less than trasmission through the female gamete.     

A gene for hybrid necrosis in an inbred line of rye (Secale cereale L.)

Summary An inbred line of rye (Secale cereale L.) has been found to carry a gene for hybrid necrosis. This gene was detected in crosses with a highly crossable wheat (Triticum aestivum L.) genotype which carries the gene Ne₂. This appears to be the first report of a gene for hybrid necrosis being present in the rye genome.     

Genetic factors influencing the regeneration ability of rye (Secale cereale L.). II. Immature embryos

Summary Immature embryos of seven rye inbred lines were cultured on modified MS medium containing 3 mg/dm^–3 2,4-D. According to thein vitro response lines were divided into four groups: (1) those producing non-embryogenic callus (NEC) from above 30% of the embryos and having a high rate of non-responding (NR) embryos, (2) those producing non-embryogenic callus (NEC) from the majority of embryos, (3) those producing NEC by the majority of embryos with a high percentage of calli regenerating roots, (4) those producing embryogenic callus (EC) and regenerating plants by above 50% of the embryos. The inheritance of these response types was analysed in F₁, F₂, and F₃ generations of crosses of some lines. The results obtained indicate that EC production and both plant and root regeneration are determined by recessive genes whereas the reduced ability for NEC production most probably by dominant genes. The lack of response is controlled by at least two interacting genes.     

Stability of RAPD markers for determining cultivar specific DNA profiles in rye (Secale cereale L.)

Summary Cultivar specific DNA profiles in rye were revealed by polymerase chain reaction (PCR) using randomly amplified polymorphic DNA (RAPD) sequences. Ten base primers were used for the amplification of genomic DNA of rye cultivars by PCR. RAPD analysis was found to be reproducible among samples between PCR runs. When amplification profiles of different rye cultivars were compared using various primers, the overall profiles were cultivar specific. However, not all primers revealed polymorphisms. These primers appear to amplify conserved sequences in all rye cultivars. Intracultivar studies were conducted on two of the cultivars. In the cultivar Imperial, no polymorphisms were observed among ten plants analyzed with five primers. In the cultivar Balboa, polymorphisms were observed among fifty plants with four of the ten primers analyzed. Despite the small amount of intracultivar variability, RAPD analysis has the potential to be a rapid and reliable method of cultivar identification in this outcrossing species.     

Effects of competition and selection pressure on yield response in winter rye (Secale cereale L.)

Summary Three intensities of mass honeycomb selection (14.3,5.3 and 1.6%) applied to an unselected rye population gave respectively an annual yield response of 0.28, -3.69 and -5.20% at 15 cm spacing, and of 4.07, 5.39 and 8.99% at 90 cm spacing. The negative response with competition was explained by strong negative correlation between competing and yielding ability which causes positive skewness because of transposition of low yielders and strong competitors from the left to the right tail of the distribution. The positive response in the absence of competition was mainly due to the increased genotypic differentiation which allowed effective discrimination between high and low yielding genotypes. The efficiency of the selection in the absence of competition was further improved by using the honeycomb designs which adjust soil heterogeneity and application of very high family and individual selection pressures.Two cycles of mass honeycomb selection increased the population yield by 29.4%, one cycle of mass plus one cycle of pedigree honeycomb selection did so by 34.5%. The results are discussed in relation to the selection response and to the efficiency of various breeding schemes.Part of senior author's doctoral thesis     

The crossing of common wheat (Triticum aestivum L.) with cultivated rye (Secale cereale L.). II. Fertilization and early post-fertilization developments

Summary Fertilization and early seed development was studied in the variety Chinese Spring of common wheat (Triticum aestivum L.) after pollination with rye (Secale cereale L.) and selfing, and in the common wheat variety Hope after selfing. In all three combinations the first pollen tube reached the micropyle in about 40 min after pollination. When pollinated with rye the migration of the sperm nuclei to the egg cell and the polar nuclei was delayed by about an hour. In the subsequent development until 72 hours after pollination the average cellular and nuclear doubling times of embryo and endosperm were about 16 and 8 hours for the interspecific combination. 18 and 9 hours for Chinese Spring selfed and 20 and 12 hours for Hope selfed.     

The quantitative relationship between telomeric heterochromatin and secalin synthesis in Secale cereale L.

Summary The relationship between the amount of prolamine (secalin) synthesized and the amount of telomeric heterochromatin in ten inbred lines of rye (Secale cereale L.) cv. Self Fertile Spring was investigated. There was no association between the total amount of secalin synthesized and the amount of telomeric heterochromatin per haploid genome. However, a positive relationship was found between total secalin synthesis and the amount of telomeric heterochromatin on the short arm of chromosome 7 (7RS). Lines synthesizing high levels of secalin were found to have approximately 60% less telomeric heterochromatin on chromosome arm 7RS than lines synthesizing low levels of secalin. There was no positive relationship between secalin synthesis and the amount of telomeric heterochromatin present on any other telomere.     

Expression of 17 rye (Secale cereale L.) traits in a range of durum wheat (Triticum durum Desf.) and bread wheat (T. aestivum L.) genetic backgrounds

Summary Expression of 17 rye traits in 24 bread wheat x rye and 8 durum wheat x rye crosses was studied, using a self-compatible, homozygous, dwarf rye. Rye showed epistasis for hairiness on the peduncle in all the crosses of Triticum aestivum and T. durum wheats with rye. Dark greenness of leaves of rye was expressed in all the durum wheat x rye and in some of the bread wheat x rye crosses. Similarly, absence of auricle pubescence, a rye trait, was expressed in most of the durum wheat x rye crosses but not in the bread wheat x rye crosses, indicating the presence of inhibitors for these traits frequently on the D genome and rarely on the A and/or B genome of wheat. Most of the wide hybrids resembled rye fully or partially for intense waxy bloom on the leaf-sheath and for the absence of basal underdeveloped spikelets. Similarly, most of the amphihaploids resembled rye for the anthocyanin in the coleoptile, stem and node. The presence of some inhibitors on A and/or B genome of wheat was indicated in some of the wheat genotypes for the expression of rye traits viz. intense waxy bloom, anthocyanin in node and absence of basal underdeveloped spikelets. Enhancement in the level of expression of the intensity and length of bristles on the mid-rib of the glume of the hybrids might be due to wheat-rye interaction. Less number of florets/spikelet as in rye showed variable expression in different wheat backgrounds. Some other rye traits like absence of auricles, terminal spikelet and glume-awn were not expressed in the wheat background. The expression of some of the rye genes might have been influenced by their interaction with Triticum cytoplasm and/or the environment.     

Identification of quantitative trait loci (QTLs) for the characters of vascular bundles in peduncle related to indica-japonica differentiation in rice (Oryza sativa L.)

The number of vascular bundles in peduncle and the ratio of vascular bundles to primary rachis branches (V/R ratio)distinguishable between indica andjaponica, are the traits associated with the processes of differentiation between indica and japonica inrice (Oryza sativa L.). In this paper a doubled-haploid population derived from the F₁ hybrid of a cross between anindica cultivar and a japonicacultivar was used to map quantitative trait loci(QTLs) controlling numbers of vascular bundles in peduncle, primary rachis branches and the V/R ratio. For vascular bundles, three QTLs were detected and they collectively explained 58.8% of the total variation. Among them, the QTLqVB-8 with the largest effect,located on chromosome 8, individually accounted for 31.1% of the total variation. Two QTLs controlling primary rachis branches, located on chromosome 8and 10 respectively, were identified and they individually explained 10.5% and18.0% of the total variation respectively. Three QTLs for the V/R ratio, mapped on chromosome 1, 2 and 8, respectively,jointly explained 61.3% of the total variation. Of the three QTLs, the QTL qV/R-1 with the largest additive effect,explained 25.3% of the total variation,was located on chromosome 1 and found to be closely linked to the gene sh-2, a major gene underlying grain-shattering ability. In addition, four and two pairs of significant epistatic QTLs were detected for vascular bundles and the V/R ratio,respectively, but none for rachis branches. Our results suggested that the numbers of vascular bundles and primary rachis branches were independently controlled by different polygenic systems, but the two polygenic systems shared a fraction of quantitative trait loci. The present study also demonstrated that the chromosome region carrying the QTL qV/R-1 for the V/R ratio and the gene sh-2 might play an important role in the processes ofindica-japonica differentiation in rice (Oryza sativa L.). This revised version was published online in August 2006 with corrections to the Cover Date.     

Detection of quantitative trait loci in Pinus sylvestris L. across years

In an earlier study, we reported the characterisation of quantitative trait loci (QTLs) for economically important traits related to wood production for a specific year of measurement. However, validating the detected QTLs across years is important for any strategy for marker-assisted selection (MAS). Therefore, we evaluated the consistency of the QTLs across four years in 9 to 12 year-old trees. Data related to tree height were analysed in two ways. The total height was used to characterise QTLs likely to be useful in MAS, and the increment values to evaluate growth regulation. Among the 11 QTLs detected, 4 were found in both the 1996 and 1997 evaluations. QTL numbers and locations related to total height were found to be highly stable. However, the QTL effects progressively changed in successive years, following either an increasing, decreasing or curvilinear trend. Depending on their specific trends, some QTLs may not be expressed at all later in maturity, or they may be expressed at a higher level. This will have direct consequences on the applicability of molecular markers in early screening programs, since a study at the mature stage may also be required. More variation in QTL number and effect was observed relating to height increment than to total height, indicating significant differences in gene expression during the growth periods involved. This revised version was published online in August 2006 with corrections to the Cover Date.     

An extended genetic map of rye (Secale cereale L.)

A genetic linkage map of rye consisting of 92 markers was constructed by using isozyme and molecular marker techniques. For this purpose an F₂ population of 137 individuals was established on which RFLP studies with homologous and heterologous probes were performed. After establishing a reliable polymerase chain reaction (PCR) protocol, 280 random primers were screened for polymorphisms and 17 random amplified polymorphic DNA (RAPD) loci were mapped. The digestion of the template DNA prior to PCR increased the degree of polymorphism. Previously published markers could also be integrated into this map by using the JoinMap computer program. The resulting linkage map comprises a total of 127 markers and spans a distance of about 760 cM.     

Isozyme electrophoretic patterns as a tool to characterize and classify rye (Secale cereale L.) seed samples

Summary Polyacrylamide gel electrophoresis of three isozymic systems (Cathodal Peroxidase, Alkaline Phosphatase and Alpha-amylase) was used to characterize and classify six rye (Secale cereale L.) seed samples. The different electrophorectic patterns allowed us to distinguish each sample. Similarities between seed samples were calculated using the Simple Matching Coefficient index (S_sm, Sneath & Sokal, 1973). High similarity values were found, and their possible reasons are discussed. Advantages of phenetic classification for plant breeders are presented.     

Mapping quantitative trait loci for yield-related traits in soybean (Glycine max L.)

Development of soybean cultivars with high seed yield is a major focus in soybean breeding programs. This study was conducted to identify genetic loci associated with seed yield-related traits in soybean and also to clarify consistency of the detected QTLs with QTLs found by previous researchers. A population of 135 F_2:3 lines was developed from a cross between a vegetable soybean line (MJ0004-6) and a landrace cultivar from Myanmar ({"type":"entrez-nucleotide","attrs":{"text":"R18500","term_id":"772110","term_text":"R18500"}}R18500). They were evaluated in the experimental field of Kasetsart University, Kamphaeng Saen, Nakhon Pathom, Thailand in a randomized complete block design with two replications each in 2011 and 2012 growing seasons. The two parents exhibited contrasting characteristics for most of the traits that were mapped. Analysis of variance showed that the main effects of genotype and environment (year) were significant for all studied traits. Genotype by environment interaction was also highly significant for all the traits. The population was genotyped by 149 polymorphic SSR markers and the genetic map consisted of 129 SSR loci which converged into 38 linkage groups covering 1156 cM of soybean genome. There were 10 QTLs significantly associated with seed yield-related traits across two seasons with single QTLs explaining between 5.0% to 21.9% of the phenotypic variation. Three of these QTLs were detected in both years for days to flowering, days to maturity and 100 seed weight. Most of the detected QTLs in our research were consistent with earlier QTLs reported by previous researchers. However, four novel QTLs including SF1, SF2 and SF3 on linkage groups L and N for seed filling period and PN1 on linkage group D1b for pod number were identified in the present study.     

The crossing of common wheat (Triticum aestivum L.) with cultivated rye (Secale cereale L.). I. crossability,pollen grain germination and pollen tube growth

Summary The crossing of common wheat (Triticum aestivum L.) with rye (Secale cereale L.), and especially the action of the crossability genes of wheat, was studied using the readily crossable wheat cv Chinese Spring (genotype kr ₁ kr ₁ kr ₂ kr ₂ >), the poorly crossable wheat cv Hope (genotype Kr ₁ Kr ₁ Kr ₂ Kr ₂ ), as well as the disomic substitution line of chromosome 5B of Hope into Chinese Spring (CS/Hope 5B, genotype Kr ₁ Kr ₁ kr ₂ kr ₂ ). By comparing crossability and actual fertilization, the poor crossability with rye of both cv Hope and the CS/Hope 5B substitution line was shown to result from absence of fertilization. Studies of pollen grain germination and pollen tube growth showed that the dominant alleles of the crossability genes manifested themselves through retardation and eventually inhibition of pollen tube growth at the style base and in the ovary wall. In Hope the growth of all pollen tubes was inhibited, whereas in CS/Hope 5B rarely fertilization was achieved. The recessive alleles of the crossability genes do not seem to have an influence on the growth of rye pollen tubes in wheat pistils.     

Molecular mapping of quantitative trait loci for ascochyta blight and botrytis grey mould resistance in an inter-specific cross in chickpea (Cicer arietinum L.) using genotyping by sequencing

Ascochyta blight (AB) and botrytis grey mould (BGM) are the most devastating fungal diseases of chickpea worldwide. The wild relative of chickpea, C. reticulatum acc. ILWC 292 was found resistant to BGM whereas, GPF2 (Cicer arietinum L.) is resistant to AB. A total of 187 F₈ Recombinant Inbred Lines (RILs) developed from an inter-specific cross of GPF2 × C. reticulatum acc. ILWC 292 were used to identify quantitative trait loci (QTLs) responsible for resistance to AB and BGM. RILs along with parents were evaluated under artificial epiphytotic field/laboratory conditions for two years. Highly significant differences (P < 0.001) were observed for reaction to both pathogens in both years. Parents and RILs were genotyped-by-sequencing to identify genome wide single nucleotide polymorphism (SNPs). A total of 1365 filtered and parental polymorphic SNPs were used for linkage map construction, of which, 673 SNPs were arranged on eight linkage groups. Composite interval mapping revealed three QTLs for AB and four QTLs for BGM resistance. Out of which, two QTLs for AB and three QTLs for BGM were consistent in both years. These QTLs can be targeted for further fine mapping for deployment of resistance to AB and BGM in elite chickpea cultivars using marker-assisted-selection.     

Identification and mapping of a quantitative trait locus controlling extreme late bolting in Chinese cabbage (Brassica rapa L. ssp. pekinensis syn. campestris L.) using bulked segregant analysis

DNA markers linked to a locus controlling an extreme late bolting trait, which was originally found in a local cultivar of a non-heading leafy vegetable,‘Osaka Shirona Bansei’ (Brassica rapa L. ssp. pekinensis syn. campestris L.) were identified using bulked segregant analysis. A doubled haploid (DH) line, DH27, which is a progeny of ‘Osaka Shirona Bansei’, shows extreme late bolting, and bolts without vernalization. DH27 was crossed with a normal bolting DH line, G309. The plantlets of the parents, F₁ and F₂, were vernalized and then grown in a greenhouse. The bolting time of F₂ plants showed a continuous distribution from 19 to 231 days after vernalization (DAV), suggesting the effects of a few major genes and polygenes. Possible linkage markers for this trait were screened by modified bulked segregant analysis (BSA). The BSA using four bulks suggested that a 530-bp RAPD band RA1255C was linked to a locus controlling the bolting trait. The RAPD band was cloned and used as a probe to detect RFLP. The fragment detected a single locus, BN007-1,the segregation of which in the F₂ population matched that of RA1255C. Three other RAPDs were found to be linked to BN007-1. A quantitative trait locus(QTL) affecting the bolting time was detected around BN007-1 using MAPMAKER/QTL. Since the difference between bolting times of both the parental genotypes in the F₂ was 138 days, these markers may be useful for a marker-assisted selection (MAS) in the breeding program for late bolting or bolting-resistant cultivars in B. rapa crops. This revised version was published online in July 2006 with corrections to the Cover Date.     

Genetic mapping of quantitative trait loci for fiber quality and yield trait by RIL approach in Upland cotton

The improvement of cotton fiber quality has become more important because of changes in spinning technology. Stable quantitative trait loci (QTLs) for fiber quality will enable molecular marker-assisted selection to improve fiber quality of future cotton cultivars. A simple sequence repeat (SSR) genetic linkage map consisting of 156 loci covering 1,024.4 cM was constructed using a series of recombinant inbred lines (RIL) developed from an F₂ population of an Upland cotton (Gossypium hirsutum L.) cross 7235 × TM-1. Phenotypic data were collected at Nanjing and Guanyun County in 2002 and 2003 for 5 fiber quality and 6 yield traits. We found 25 major QTLs (LOD ≥ 3.0) and 28 putative QTLs (2.0 < LOD < 3.0) for fiber quality and yield components in two or four environments independently. Among the 25 QTLs with LOD ≥ 3, we found 4 QTLs with large effects on fiber quality and 7 QTLs with large effects on yield components. The most important chromosome D8 in the present study was densely populated with markers and QTLs, in which 36 SSR loci within a chromosomal region of 72.7 cM and 9 QTLs for 8 traits were detected.     

Identification of quantitative trait loci for grain yield, seed protein concentration and maturity in field pea (Pisum sativum L.)

Breeding efforts to improve grain yield, seed protein concentration and early maturity in pea (Pisum sativum L.) have proven to be difficult. The use of molecular markers will improve our understanding of the genetic factors conditioning these traits and is expected to assist in selection of superior genotypes. This study was conducted to identify genetic loci associated with grain yield, seed protein concentration and early maturity in pea. A population of 88 recombinant inbred lines (RILs) that was developed from a cross between 'Carneval' and 'MP1401' was evaluated at 13 environments across the provinces of Alberta, Manitoba and Saskatchewan, Canada in 1998, 1999 and 2000. A linkage map consisting of 193 AFLPs (amplified fragment length polymorphism), 13 RAPDs (random amplified polymorphic DNA) and one STS (sequence tagged site) marker was used to identify putative quantitative trait loci (QTL) for grain yield, seed protein concentration and early maturity. Four QTL were identified each for grain yield and days to maturity, and three QTL were identified for seed protein concentration. A multiple QTL model for each trait showed that these genomic regions accounted for 39%, 45% and 35% of the total phenotypic variation for grain yield, seed protein concentration and days to maturity, respectively. The consistency of these QTL across environments and their potential for marker-assisted selection are discussed in this report.     

Variation in heading date conceals quantitative trait loci for other traits of importance in breeding selection of rice

To identify quantitative trait loci (QTLs) associated with the primary target traits for selection in practical rice breeding programs, backcross inbred lines (BILs) derived from crosses between temperate japonica rice cultivars Nipponbare and Koshihikari were evaluated for 50 agronomic traits at six experimental fields located throughout Japan. Thirty-three of the 50 traits were significantly correlated with heading date. Using a linkage map including 647 single-nucleotide polymorphisms (SNPs), a total of 122 QTLs for 38 traits were mapped on all rice chromosomes except chromosomes 5 and 9. Fifty-eight of the 122 QTLs were detected near the heading date QTLs Hd16 and Hd17 and the remaining 64 QTLs were found in other chromosome regions. QTL analysis of 51 BILs having homozygous for the Koshihikari chromosome segments around Hd16 and Hd17 allowed us to detect 40 QTLs associated with 27 traits; 23 of these QTLs had not been detected in the original analysis. Among the 97 QTLs for the 30 traits measured in multiple environments, the genotype-by-environment interaction was significant for 44 QTLs and not significant for 53 QTLs. These results led us to propose a new selection strategy to improve agronomic performance in temperate japonica rice cultivars.