S‐locus diversity and cross‐compatibility of wild Prunus avium for timber breeding

Prunus avium is primarily cultivated for its fruit, sweet cherries. However, it is also used to produce high‐quality timber. In a P. avium seed orchard, gametophytic self‐incompatibility is a restriction for free pollen flow and should be considered when establishing basic forest materials. In this study, S‐locus diversity and cross‐incompatibility of wild cherry individuals in clonal banks established for breeding for timber production were investigated. Wild cherry trees (140) with outstanding forest growth habit, collected in northern Spain, grafted and planted in two clonal banks, were genotyped at the S‐locus. The self‐incompatibility S‐locus genes, S‐RNase and SFB, were analysed by PCR. Twenty‐two S‐haplotypes, resulting in 72 different S‐genotypes, were identified. The genotypes were grouped into 33 incompatibility groups and 39 unique genotypes. This initial S‐locus analysis revealed large genetic diversity of wild cherry trees from the Spanish northern deciduous forest, and provides useful information for seed orchard design. Wild P. avium displays significantly more genetic diversity than what is detected in local cultivars, revealing a narrowing of genetic diversity during local domestication.     

PPR‐B‐31: a new maintainer allele in the male‐fertility restorer gene of radish (Raphanus sativus) Ogura cytoplasm

The PPR‐B gene is responsible for male‐fertility restoration of the Ogura‐type male‐sterile radish plants, and it is located in the complex Rfo locus in the vicinity of similar PPR‐A gene and PPR‐C pseudogene. The aim of this study was to identify PPR‐B alleles and understand the structure of the Rfo locus in radish breeding lines. Five lines of radish with normal male‐fertile cytoplasm were tested. The entire PPR‐B gene was amplified, sequenced and allelic PPR‐B sequences were identified. The results indicated that the maintainer lines 7, 15 and 21 contained a non‐restoring form of PPR‐B protein. A unique PPR‐B was found in lines 24/15 and 31 that are restorer and maintainer lines, respectively. The substitutions might be responsible for the loss of a restoring function of the PPR‐B‐31 allele. Amplification of the PPR‐A/PPR‐B and PPR‐B/PPR‐C intergenic regions allowed to identify rearrangements within Rfo locus. Obtained results confirm the wide allelic variation within the Rfo locus, as well as high genetic complexity of the fertility restoration mechanism in radish.     

In vitro induction of tetraploids in colchicine-treated cocoyam plantlets

A self-compatible (SC) variant of a wild diploid potato species, Solanum chacoense, which is normally self-incompatible (SI), was investigated for the nature and genetics of self-compatibility. It was crossed with a SI cultivated diploid potato species, S. phureja. The F₁ progeny segregated SC vs. SI. Diallel crosses were made among 15 F₁'s. Self-compatibility was tested in a selfed family of a parental SC variant and in sib-mated and selfed families of F₁ progeny. All the data suggest that there is a single dominant gene (Sli) with sporophytic action inhibiting S gene expression in the pollen. Plants having a ‘Sli’ gene, produce pollen which is compatible to its own parent and plants with similar S genes. The ‘Sli’ gene has been maintained in a heterozygous condition through eight selfing generations (S₈) implying that dominant homozygotes might be associated with lethality. This revised version was published online in July 2006 with corrections to the Cover Date.     

Fine mapping the QTL qFS‐chr.23, using a CSIL

In an earlier advanced‐backcross quantitative trait locus (QTL) analysis of an interspecific cross of Gossypium hirsutum cv. ‘Xinluzhong 36’(‘XLZH36’) and G. barbadense cv. ‘Xinhai 21’(‘XH21’), a QTL for fibre strength in the chromosome segment introgression line IL23‐09 was analysed. Single marker analysis revealed that the markers on chro.23 were associated with fibre strength. Using composite interval mapping with the F₂ population (1296 plants), a QTL for fibre strength was detected on chro. 23. The QTL explained 8.9% and 15.9% of phenotypic variances in the F₂ and F_2 : 3 generations, respectively. Substitution mapping suggested that the QTL was located at a physical distance of 23.4 kb between the markers BNL1414 and the single nucleotide polymorphism (SNP) locus D09_43776813 C‐G. We designated this QTL as qFS‐chr.23 (quantitative trait locus for fibre strength on chro.23). This work provides a valuable genetic resource for the breeding of high fibre quality in cotton and will facilitate future efforts for map‐based cloning.     

Effects of the Gpc‐B1 locus on high grain protein content introgressed into Argentinean wheat germplasm

Wheat grain protein content (GPC) is important for human nutrition and has a strong influence on the quality of pasta and bread. The objective of this study was to analyse the introduction of the Gpc‐B1 allele into two Argentinean bread wheat cultivars. Near‐isogenic lines were developed in ‘ProINTA Oasis’ and ‘ProINTA Granar’ using marker‐assisted selection. Gpc‐B1 lines showed a significant (P = 0.01) increase in GPC and a significant (P = 0.001) decrease in grain weight in comparison with control lines without Gpc‐B1. Differences in yield were not significant (P = 0.49) between lines. Gpc‐B1 lines significantly reduced (P = 0.02) straw nitrogen concentration at maturity and significantly increased (P = 0.02) the nitrogen harvest index. When data were analysed by genotype and environment, differences in some analysed parameters were found, indicating that Gpc‐B1 expression may be affected by different genetic backgrounds and environmental conditions. These results suggest that the introgression of the Gpc‐B1 allele into Argentinean wheat germplasm could be a valuable resource for improving GPC with no detrimental effect on grain yield.     

Prediction of deoxynivalenol and zearalenone concentrations in Fusarium graminearum inoculated backcross populations of maize by symptom rating and near‐infrared spectroscopy

Gibberella ear rot (GER) caused by Fusarium graminearum is a destructive disease in maize of temperate regions resulting in yield reduction and contamination by the mycotoxins deoxynivalenol (DON) and zearalenone (ZON). We wanted to analyse whether prediction of DON and ZON concentrations is feasible either by GER severity ratings or by near‐infrared spectroscopy (NIRS). We analysed 80 and 102 lines developed by backcrossing doubled‐haploid lines from segregating populations to the resistant and susceptible parent, respectively, by artificial infection at three locations in Germany and France. Both backcross (BC) populations differed substantially in their means for all traits with significant (P < 0.01) genotypic variances. DON and ZON concentrations measured by immunotests were significantly (P < 0.01) correlated with each other and with GER severity within each BC population (0.6 ≤ r ≤ 0.9, P < 0.01). DON concentration measured by immunotest and NIRS significantly correlated (r ≈ 0.9, P < 0.01). In conclusion, DON and ZON concentrations could be reliably predicted by GER severity. Additional NIRS analysis of DON concentration might be useful for the positively selected fraction.     

Molecular mechanism of the S-RNase-based gametophytic self-incompatibility in fruit trees of Rosaceae

Self-incompatibility (SI) is a major obstacle for stable fruit production in fruit trees of Rosaceae. SI of Rosaceae is controlled by the S locus on which at least two genes, pistil S and pollen S, are located. The product of the pistil S gene is a polymorphic and extracellular ribonuclease, called S-RNase, while that of the pollen S gene is a protein containing the F-box motif, SFB (S haplotype-specific F-box protein)/SFBB (S locus F-box brothers). Recent studies suggested that SI of Rosaceae includes two different systems, i.e., Prunus of tribe Amygdaleae exhibits a self-recognition system in which its SFB recognizes self-S-RNase, while tribe Pyreae (Pyrus and Malus) shows a non-self-recognition system in which many SFBB proteins are involved in SI, each recognizing subset of non-self-S-RNases. Further biochemical and biological characterization of the S locus genes, as well as other genes required for SI not located at the S locus, will help our understanding of the molecular mechanisms, origin, and evolution of SI of Rosaceae, and may provide the basis for breeding of self-compatible fruit tree cultivars.     

Prezygotic interspecific hybridization barriers between kenaf (Hibiscus cannabinus L.) and four wild relatives

Use of interspecific hybridization in genetic improvement of kenaf (Hibiscus cannabinus), a multipurpose crop, is limited by low cross‐compatibility. Previous hybridization studies suggest that part of incompatibility barrier is prezygotic. Pollen germination, stigma receptivity and growth of alien pollen on kenaf pistil were studied using wild relatives of kenaf. Four wild and semi‐domesticated species, H. surattensis, H. acetosella, H. radiatus and H. vitifolius, were used as pollen donors in the present experiment. H. radiatus exhibited highest pollen germination (65.89%) on H. cannabinus stigma. For closely related species (H. surattensis, H. acetosella, H. radiatus), stylar incompatibility was noticed, while for distantly related species H. vitifolius, the incompatibility was stigmatic. Number of pollen tubes entering ovule of H. cannabinus pistil was higher for H. radiatus (1.30). Alien pollen growth and penetration of pollen tube through ovule were higher in species sharing same genome of H. cannabinus. High callose deposition was observed in incompatible crosses, suggesting a general mechanism for prezygotic incompatibility in Hibiscus section Furcaria. Callose deposition in incompatible crosses increased with time.     

Association analysis of five candidate genes with plant height and dry matter yield in perennial ryegrass

Five candidate genes LpIAA1, LpRUB1, LpBRI1, LpSHOOT1 and LpTB1 with putative function in plant architecture were allele sequenced in a panel of 96 diploid perennial ryegrass genotypes of diverse origin. The total length of the non‐redundant genomic DNA alignment was 5425 bp and revealed 270 polymorphic sites in total. A negative significant Tajima's D value was detected in LpTB1 gene, suggesting selection pressure for low‐frequency alleles in this gene. All 96 genotypes were evaluated for plant height and dry matter yield over two years. Marker–trait associations were calculated between polymorphic sites and both phenotypic traits. Three indels and three single nucleotide substitutions in LpTB1 gene were significantly (P < 0.05, q < 0.05) associated with plant height, while one indel was associated with dry matter yield. The results suggest putative role of LpTB1 gene in plant height determination in perennial ryegrass and provide means for target allele selection.     

Development of a molecular marker for self‐compatible S4′ haplotype in sweet cherry (Prunus avium L.) using high‐resolution melting

Sweet cherry (Prunus avium L.) has stylar gametophytic self‐incompatibility, which is controlled by the multi‐allelic S‐locus and encompasses the highly polymorphic genes for the S‐ribonuclease (S‐RNase) and S‐haplotype‐specific F‐box (SFB), which are female and male determinants, respectively. The self‐compatible mutant SFB4′ corresponds to an allele variant of SFB4 and presents a frameshift mutation. Even though male‐determinant molecular markers can discriminate between SFB4 and SFB4′ alleles, the methods required are laborious, time‐consuming and expensive, and not suitable for massive analysis and integration into breeding programmes. Our aim was to develop molecular markers for the evaluation of self‐compatibility alleles in sweet cherry, that could be used as a high‐throughput screening strategy to identify SFB4 and SFB4′ alleles, based on a marker for male determinacy. Our results were consistent using primers flanking the mutation responsible for the SFB4′ allele. We designed a specific molecular marker and confirmed it in sweet cherry commercial varieties. This new molecular marker is feasible for self‐compatibility alleles in the male determinant in sweet cherry‐assisted breeding programs.     

QTL epistatic analysis for yield components with single‐segment substitution lines in rice

Epistasis is an important genetic component in determining the phenotype of complex quantitative trait. In this article, 12 single‐locus heterozygotes and 66 double‐locus heterozygotes were developed and then were applied to assay QTL epistasis for four yield‐associated traits under two planting densities. Of 264 (66 × 4) tested interactions, 130 (49.2%) were significant at the p < .05 level. QTL with the same effect directions had higher probabilities of interactions. The negative epistasis at least included one positive effect QTL but the positive epistasis one negative QTL. The detected epistasis was sensitive to planting density. Epistasis also exhibited pleiotropic effects.     

Distribution of S haplotypes in commercial cultivars of Brassica rapa

Self‐incompatibility in Brassicaceae plants is sporophytically controlled by a single multi‐allelic locus (S locus), which contains at least three highly polymorphic genes expressed in the stigma (SLG and SRK) and in the pollen (SCR/SP11). Using polymerase chain reaction‐restriction fragment length polymorphism (PCR‐RFLP) analysis with SXG‐specific primer pairs, the S haplotypes of F₁ hybrid and open‐pollinated commercial cultivars of Brassica rapa were identified. The number of S haplotypes detected in the F₁ hybrid cultivars of Chinese cabbage, komatsuna, pak‐choi, turnip, open‐pollinated cultivars of Chinese cabbage and turnip were 9, 9, 4, 11, 13 and 12, respectively. Nine of them had different PCR‐RFLP profiles from those of the S‐tester lines that determined the SLG sequences. Four SLG sequences in the F₁ hybrid cultivars were determined and named S⁵³, S⁵⁴, S⁵⁵ and S⁵⁶, respectively. It is demonstrated that the PCR‐RFLP analysis using specific primer pairs of SLG and SRK is useful for identification of the S haplotypes, in both, S homozygous and S heterozygous plants of B. rapa. The possibility of using this method routinely in breeding programmes, and in the evaluation of F₁ hybrid seed purity, is discussed.     

SFB‐based S‐haplotyping of apricot (Prunus armeniaca) with DHPLC

Most cultivars that belong to the Rosaceae are self‐incompatible and depend on cross‐pollination. The pollen donor and pollen recipient have to flower synchronously and must be genetically compatible. Genetic compatibility is governed by the S‐locus, which holds the S‐RNase and S‐haplotype‐specific F‐Box (SFB) genes. Thus, the S‐genotype of cultivars is an important feature and is characterized molecularly by the S‐RNase and SFB alleles which are distinctive for each S‐haplotype. Here, we report the usage of DNA chromatography (denaturing high‐performance liquid chromatography – DHPLC) for identifying the S‐genotypes of European apricots on the basis of their SFB alleles. DHPLC is amenable to high‐throughput automation, and therefore is valuable for breeding and for high‐quality plant typing in the nursery.     

Genetics of self-compatibility in a self-incompatible wild diploid potato species Solanum chacoense. 2. Localization of an S locus inhibitor (Sli) gene on the potato genome using DNA markers

A self-compatible (SC) hybrid plant F₁-1 was obtained from a cross between a SC variant of a wild diploid potato species, Solanum chacoense, and a self-incompatible (SI) cultivated diploid species, S. phureja. The clone F₁-1 has previously been proposed to have a dominant S locus inhibitor gene (Sli) in a heterozygous condition. It was crossed as a male parent with a selected clone from a S. stenotomum-S. phureja population, resulting in a segregating population consisting of 116 hybrid plants. Self-compatibility was assessed by selfing each of the hybrids. Sixty-six of them were SC, while 35 were SI, showing a significant distortion from an expected Mendelian ratio of 1:1. A genetic linkage map was constructed using DNA markers to localize the Sli gene. A total of 28 RAPD and 127 RFLP markers identified 109 mapping positions on 12 linkage groups. The Sli gene was mapped at a distal end of chromosome 12. Since the S locus has been localized on chromosome 1 on the potato RFLP map, it is confirmed that the Sli gene is independent of the S locus. This revised version was published online in August 2006 with corrections to the Cover Date.     

A statistical mixture model for estimating the proportion of unreduced pollen grains in perennial ryegrass (Lolium perenne L.) via the size of pollen grains

Summary The size of pollen grains is commonly used to indicate the ploidy level of pollen grains. In this paper observations of the diameter of pollen grains are evaluated from one diploid accession of perennial ryegrass (Lolium perenne L.), which was expected to produce diploid (unreduced) pollen grains in addition to haploid pollen grains. The considerable overlap of the diameter distributions of haploid and diploid pollen grains severely hampers the accurate estimation of the proportion of diploid pollen grains. To overcome this problem we develop in this paper a statistical normal mixture model and we describe a method to test for the production of diploid pollen grains from a diploid parent, and to estimate the proportion of diploid pollen grains.     

Compatibility and incompatibility in witloof-chicory (Cichorium intybus L.). 2. The incompatibility system

Summary In witloof-chicory fertilization is frequently prevented by the action of an incompatibility mechanism. In two experiments the incompatibility system was investigated. In the IVT phytotron at a constant temperature of 17 C reciprocal crosses were made between unrelated plants followed by diallel crosses between the F1 plants and backcrosses with the parents.Analyses of the results of the above crosses strongly suggest that in witloof-chicory a one locus sporophytic incompatibility system occurs with different dominance and codominance relationships between S-alleles in pollen and style.     

1‐Methylcyclopropene (1‐MCP)‐Induced Alteration in Leaf Photosynthetic Rate,Chlorophyll Fluorescence,Respiration and Membrane Damage in Rice (Oryza sativa L.) Under High Night Temperature

High night temperature (HNT) can induce ethylene‐triggered reactive oxygen species production, which can cause premature leaf senescence and membrane damage, thereby affecting production, consumption and transfer of photosyn‐thates, and yield. The 1‐methylcyclopropene (1‐MCP) can competitively bind with ethylene receptors and decrease ethylene effects. The objective was to determine the effects of HNT and 1‐MCP on leaf photosynthetic rate (P_N), chlorophyll fluorescence, total chlorophyll (TC), respiration, membrane damage, pollen germination, spikelet fertility (SF) and yield of rice hybrid ‘XL723’. Plants were grown under ambient night temperature (ANT) (25 °C) or HNT (30 °C) with or without 1‐MCP treatment. Application of 1‐MCP was at the boot stage. The decrease in yield (11 %) under HNT was associated with decreased P_N (4 %), stomatal conductance (8 %), quantum yield (11 %) TC (23 %) and SF (5 %) and increased respiration (74 %), F_o/F_m (increase in thylakoid membrane damage; 11 %) and membrane damage (leaf electrolytic leakage; 57 %). The 1‐MCP‐treated plants grown under HNT showed increased yield (17 %), which was associated with increased P_N (10 %), stomatal conductance (30 %), quantum yield (9 %), TC (37 %) and SF (11 %) and decreased respiration (39 %), F_o/F_m (5 %) and membrane damage (18 %). Plants grown under HNT showed increased grain chalkiness (154 %) compared with plants grown under ANT.     

Cross compatibility between diploid and tetraploid perennial ryegrass (Lolium perenne L.)

Summary A number of crosses were made between diploid and tetraploid Lolium perenne plants in order to determine the degree of cross compatibility between the two different ploidy levels. The range of hybridization undertaken involved diploidxdiploid, and tetraploid x tetraploid control crossings, diploid x tetraploid crosses, tetraploid x diploid crosses and mixed pollinations. The seed set, the germination capacity of resultant hybrid seeds, and the chromosome constitution of derived seedlings was determined. In addition attempts were made to culture 12 day-old hybrid embryos from the diploid x tetraploid and reciprocal crosses.The crossing results obtained indicated no barrier to fertilization between diploids and tetraploids, but the irregular and poor development of the seed accompanied by much reduced germination, indicated unsatisfactory endosperm establishment of large numbers of triploids, as confirmed by the result of culturing embryos dissected from 12-day old seeds.In preliminary studies undertaken to determine the extent to which diploid and tetraploid ryegrass varieties intercross when grown in close proximity for seed production, Griffiths and Pegler (1966) observed a very low incidence of triploids amongst the progenies of diploid plants exposed to the pollen of tetraploid plants and also amongst the progenies of tetraploid material exposed to the pollen of diploid plants.In these and subsequent studies it was noted that only a very small proportion of the so-called fully formed seeds derived from diploid x tetraploid crosses did actually germinate. When, in 1964, propagules of clones of S.24 perennial ryegrass, well separated from one another, were placed in drills of the tetraploid perennial ryegrass variety Reveille, the proportion of florets which formed caryopses, and of caryopses which germinated was as follows (Table 1).The data (Table 1) clearly indicate that a considerable proportion of the S.24 florets had been fertilized by 2n pollen and had formed caryopses, but most of these had proved to be defective. As noted in the previous studies, the frequency of triploid seedlings was low, thus suggesting incompatibility between the n and 2n gametes for the production of viable zygotes.Of the total progenies classed as tetraploids in the early seedling stage, 80% proved to be ryegrass x tall fescue F₁ hybrids. These arose as a result of free crossing with     

Inheritance of resistance to the peach root‐knot nematode (Meloidogyne floridensis) in interspecific crosses between peach (Prunus persica) and its wild relative (Prunus kansuensis)

The peach root‐knot nematode, Meloidogyne floridensis (MF), infects majority of available nematode‐resistant peach rootstocks which are mostly derived from peach (Prunus persica) and Chinese wild peach (P. davidiana). Interspecific hybridization of peach with its wild relative, Kansu peach (P. kansuensis), offers potential for broadening the resistance spectrum in standard peach rootstocks. We investigated the inheritance of resistance to MF in segregating populations of peach (‘Okinawa’ or ‘Flordaguard’) × P. kansuensis. A total of 379 individuals from 13 F₂ and BC₁F₁ families were challenged with a pathogenic MF isolate “MFGnv14” and were classified as resistant (R) or susceptible (S) based on root galling intensity. Segregation analyses in F₂ progeny revealed the involvement of a major locus with a dominant or recessive allele determining resistance in progeny segregating 3R:1S and 1R:3S, respectively. Testcrosses with a homozygous‐susceptible peach genotype (‘Flordaguard’ or ‘UFSharp’) confirmed P. kansuensis as a source of new resistance and the heterozygous allelic status of P. kansuensis at the locus conferring resistance to MF. We propose a single‐locus dominant/recessive model for the inheritance of resistance.