Development of microsatellite markers in cultivated and wild species of sections <Emphasis Type="Italic">Cepa</Emphasis> and <Emphasis Type="Italic">Phyllodolon</Emphasis> in <Emphasis Type="Italic">Allium</Emphasis>

The potential of microsatellite markers for use in genetic studies has been evaluated in Allium cultivated species (Allium cepa, A. fistulosum) and its allied species (A. altaicum, A. galanthum, A. roylei, A. vavilovii). A total of 77 polymerase chain reaction (PCR) primer pairs were employed, 76 of which amplified a single product or several products in either of the species. The 29 AMS primer pairs derived from A. cepa and 46 microsatellites primer pairs from A. fistulosum revealed a lot of polymorphic amplicons between seven Allium species. Some of the microsatellite markers were effective not only for identifying an intraspecific F₁ hybrid between shallot and bulb onion but also for applying to segregation analyses in its F₂ population. All of the microsatellite markers can be used for interspecific taxonomic analyses among two cultivated and four wild species of sections Cepa and Phyllodolon in Allium. Generally, our data support the results obtained from recently performed analyses using molecular and morphological markers. However, the phylogeny of A. roylei, a threatened species with several favorable genes, was still ambiguous due to its different positions in each dendrogram generated from the two primer sets originated from A. cepa and A. fistulosum.     

Search in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm for sources of broad-spectrum resistance to four <Emphasis Type="Italic">Tospovirus</Emphasis> species

The genus Tospovirus was considered as monotypic with Tomato spotted wilt virus (TSWV) being the only assigned species. However, extensive studies with worldwide isolates revealed that this genus comprises a number of species with distinct virulence profiles. The Neotropical South America is one center of Tospovirus diversity with many endemic species. Groundnut ringspot virus (GRSV), TSWV, Tomato chlorotic spot virus (TCSV), and Chrysanthemum stem necrosis virus (CSNV) are the predominant tomato-infecting species in Brazil. Sources of resistance were found in Solanum (section Lycopersicon) mainly against TSWV isolates from distinct continents, but there is an overall lack of information about resistance to other viral species. One-hundred and five Solanum (section Lycopersicon: Solanaceae) accessions were initially evaluated for their reaction against a GRSV isolate by analysis of symptom expression and systemic virus accumulation using DAS-ELISA. A subgroup comprising the most resistant accessions was re-evaluated in a second assay with TSWV, TCSV, and GRSV isolates and in a third assay with a CSNV isolate. Seven S. peruvianum accessions displayed a broad-spectrum resistance to all viral species with all plants being free of symptoms and systemic infection. Sources of resistance were also found in tomato cultivars with the Sw-5 gene and also in accessions of S. pimpinellifolium, S. chilense, S. arcanum, S. habrochaites, S. corneliomuelleri, and S. lycopersicum. The introgression/incorporation of these genetic factors into cultivated tomato varieties might allow the development of genetic materials with broad-spectrum resistance, as well as with improved levels of phenotypic expression.     

Molecular markers linked to <Emphasis Type="Italic">Bn</Emphasis>;<Emphasis Type="Italic">rf</Emphasis>: a recessive epistatic inhibitor gene of recessive genic male sterility in <Emphasis Type="Italic">Brassica napus </Emphasis>L.

7–7365AB is a recessive genic male sterile (RGMS) two-type line, which can be applied in a three-line system with the interim-maintainer, 7–7365C. Fertility of this system is controlled by two duplicate dominant epistatic genes (Bn;Ms3 and Bn;Ms4) and one recessive epistatic inhibitor gene (Bn;rf). Therefore an individual with the genotype of Bn;ms3ms3ms4ms4Rf_ exhibits male sterility, whereas, plant with Bn;ms3ms3ms4ms4rfrf shows fertility because homozygosity at the Bn;rf locus (Bn;rfrf) can inhibit the expression of two recessive male sterile genes in homozygous Bn;ms3ms3ms4ms4 plant. A cross of 7–7365A (Bn;ms3ms3ms4ms4RfRf) and 7–7365C (Bn;ms3ms3ms4ms4rfrf) can generate a complete male sterile population served as a mother line with restorer in alternative strips for the multiplication of hybrid seeds. In the present study, molecular mapping of the Bn;Rf gene was performed in a BC₁ population from the cross between 7–7365A and 7–7365C. Bulked segregant analysis (BSA) and amplified fragment length polymorphism (AFLP) technique was used to identify molecular markers linked to the gene of interest. From a survey of 768 primer combinations, seven AFLP markers were identified. The closest marker, XM5, was co-segregated with the Bn;Rf locus and successfully converted into a sequence characterized amplified region (SCAR) marker, designated as XSC5. Two flanking markers, XM3 and XM2, were 0.6 cM and 2.6 cM away from the target gene, respectively. XM1 was subsequently mapped on linkage group N7 using a doubled-haploid (DH) mapping population derived from the cross Tapidor × Ningyou7, available at IMSORB, UK. To further confirm the location of the Bn;Rf gene, additional simple sequence repeat (SSR) markers in linkage group N7 from the reference maps were screened in the BC₁ population. Two SSR markers, CB10594 and BRMS018, showed polymorphisms in our mapping population. The molecular markers found in the present study will facilitate the selection of interim-maintainer.     

Genetic analysis reveals a dominant <Emphasis Type="Italic">S</Emphasis> locus and an <Emphasis Type="Italic">S</Emphasis> suppressor locus in natural self-compatible <Emphasis Type="Italic">Brassica napus</Emphasis>

A self-incompatible (SI) line, S-1300, and its maintainer 97-wen135, a self-compatible (SC) line, were used to study the inheritance of maintenance for self-incompatibility in B. napus. The ratio of SI plants to SC plants from S-1300 × 97-wen135 F₂ and (S-1300 × 97-wen135) × 97-wen135 was 346:260 and 249:232, fitting the expected ratio of 9:7 and 1:1, respectively. Based on these observations, here we propose a genetic model in which two independent loci, S locus and S suppressor locus (sp), are predicted to control the inheritance of maintenance for self-incompatibility in B. napus. The genotypes of S-1300 and 97-wen135 are S ₁₃₀₀ S ₁₃₀₀ sp ₁₃₀₀ sp ₁₃₀₀ and S ₁₃₅ S ₁₃₅ sp ₁₃₅ sp ₁₃₅ , respectively. S ₁₃₅ is dominant to S ₁₃₀₀ , but coexistence of sp ₁₃₀₀ and sp ₁₃₅ fails to suppress S locus. Both S ₁₃₀₀ and S ₁₃₅ can be suppressed by sp ₁₃₅ , while sp ₁₃₀₀ can suppress S ₁₃₅ but not S ₁₃₀₀ . The model contains two characteristics: that a dominant S locus exists in self-compatible B. napus, and that co-suppression will occur when sp loci are heterozygous. The model has been validated by the segregation of S phenotypes in the (S-1300 × 97-wen135) × S-1300, the progenies of SC S-1300 × 97-wen135 F₂ plants and DH population developed from S-1300 × 97-wen135 F₁. This is the first study to report co-suppression of S suppressor loci in B. napus. The genetic model will be very useful for developing molecular markers linked to maintenance for self-incompatibility and for dissecting the mechanism of SI/SC in B. napus.     

Molecular mapping of <Emphasis Type="Italic">Pc68</Emphasis>, a crown rust resistance gene in <Emphasis Type="Italic">Avena</Emphasis><Emphasis Type="Italic">sativa</Emphasis>

Crown rust, which is caused by Puccinia coronata f. sp. avenae, P. Syd. & Syd., is the most destructive disease of cultivated oats (Avena sativa L.) throughout the world. Resistance to the disease that is based on a single gene is often short-lived because of the extremely great genetic diversity of P. coronata, which suggests that there is a need to develop oat cultivars with several resistance genes. This study aimed to identify amplified fragment length polymorphism AFLP markers that are linked to the major resistance gene, Pc68, and to amplify the F₆ genetic map from Pc68/5*Starter × UFRGS8. Seventy-eight markers with normal segregation were discovered and distributed in 12 linkage groups. The map covered 409.4 cM of the Avena sativa genome. Two AFLP markers were linked in repulsion to Pc68: U8PM22 and U8PM25, which flank the gene at 18.60 and 18.83 centiMorgans (cM), respectively. The marker U8PM25 is located in the linkage group 4_12 in the Kanota × Ogle reference oat population. These markers should be useful for transferring Pc68 to genotypes with good agronomic characteristics and for pyramiding crown rust resistance genes.     

Microsporogenesis of <Emphasis Type="Italic">Rps</Emphasis>8/<Emphasis Type="Italic">rps</Emphasis>8 heterozygous soybean lines

Phytophthora root and stem rot caused by Phytophthora sojae, is one of the most damaging diseases of soybean, for which management is principally done by planting resistant cultivars with race specific resistance which are conferred by Rps (Resistance to Phytophthora sojae) genes. The Rps8 locus, identified in the South Korean landrace PI 399073, is located in a 2.23 Mbp region on soybean chromosome 13. In eight cv. Williams (rps8/rps8) × PI 399073 (Rps8/Rps8) populations, this region exhibited strong segregation distortion. In a cross between the South Korean lines PI 399073 (Rps8/Rps8) and PI 408211B (multiple Rps genes) this region segregated in a Mendelian fashion. In this study, microsporogenesis was evaluated to identify meiotic abnormalities that may be associated with the segregation distortion of the Rps8 region. Pollen was collected from greenhouse-grown plants of the parental genotypes: Williams, PI 399073, and PI 408211B; as well as selected Rps8/rps8 RILs from Williams × PI 399073 BC₄F_2:3 and PI 399073 × PI 408211B F_4:5 populations. There were no differences for pollen viability among the genotypes. However, for PI 399073, a mix of dyads, triads, tetrads and pentads was observed. A high frequency of meiotic abnormalities including fragments, laggards, multinucleated microspores; and microcytes containing DNA was also observed in Rps8/rps8 Williams × PI 399073 BC₄F_2:3 RILs. These meiotic abnormalities may contribute to the high degree of segregation distortion present in the Williams × PI 399073 populations.     

Resistance screening of <Emphasis Type="Italic">Coffea</Emphasis> spp. accessions for <Emphasis Type="Italic">Pratylenchus coffeae</Emphasis> and <Emphasis Type="Italic">Radopholus arabocoffeae</Emphasis> in Vietnam

Coffee varieties with resistance for the plant-parasitic nematodes Pratylenchus coffeae and Radopholus arabocoffeae are limited in Vietnam. A selection of imported varieties and high yield varieties of Arabica coffee in Vietnam were evaluated for resistance to both plant-parasitic nematode species in Northern Vietnam. The same experiments were carried out with hybrid arabica coffee, three selected clones of Coffea canephora and one clone of Coffea excelsa in the Western Highland of Vietnam. The screened coffee accessions from Ethiopia (KH1, KH13, KH20, KH21, KH29, and KH31) were susceptible and good host for P. coffeae. Also accessions 90P4 (Portugal) and Oro azteca (Mexico) had a reproduction factor Rf > 1. Pluma Hidalgo (Mexico), 90/6 (Vietnam), 90P3 (Portugal), 90P2 (Vietnam), Variedad (Mexico), 90T (Portugal), and Garnica (Mexico) were poor hosts (Rf < 1) but not tolerant to P. coffeae, expressed by a reduction of root weight compared to untreated control plants. Most of the coffee accessions tested in Northern Vietnam were intolerant to R. arabocoffeae, except 90T which showed no reduction of root weight, even at high initial nematode densities (4,000/pot). Good hosts for R. arabocoffeae were Variedad, KH1, KH21, KH29, KH20, KH31, and KH13 with Rf > 1. Pluma Hidalgo, 90/6, 90P3, 90P2, 90T, Oro azteca, and Garnica were poor hosts (Rf < 1). In the Western Highland experiment, all arabica coffee accessions were susceptible for P. coffeae with Rf ranging from 1.41 to 1.59. Tolerance to P. coffeae was found in C. liberica var. Dewevrei, Hong34 and Nhuantren. Coffea excelsa, Hong34, Nhuantren, and H1C19 were tolerant to R. arabocoffeae at the highest inoculation density (4,000 nematodes/pot). The most susceptible accessions were Nhuantren and K55. Resistance (Rf < 1) to R. arabocoffeae was found in C. liberica var. Dewevrei and Hong34. This article reports on the first screening for resistance and tolerance to P. coffeae and R. arabocoffeae in coffee accessions in Vietnam and shows promising results for enhanced coffee-breeding.     

Role of <Emphasis Type="Italic">Berberis</Emphasis> spp. as alternate hosts in generating new races of <Emphasis Type="Italic">Puccinia graminis</Emphasis> and <Emphasis Type="Italic">P. striiformis</Emphasis>

The common barberry and several other Berberis spp. serve as the alternate hosts to two important rust pathogens of small grains and grasses, Puccinia graminis and P. striiformis. Barberry eradication has been practiced for centuries as a means to control stem rust. Diverse virulence variations have been observed in populations of P. graminis f. sp. tritici that were associated with susceptible barberries in North America. Barberry likely has played a role in generating new races of P. striiformis f. sp. tritici in some regions in the world. Several North American stem rust races, namely races 56, 15B and QCC, initially originated from barberry, were subsequently responsible for generating large-scale epidemics. Thus, sexual cycles on Berberis spp. may generate virulence combinations that could have serious consequences to cereal crop production.     

Expression of <Emphasis Type="Italic">Bruguiera gymnorhiza</Emphasis><Emphasis Type="Italic">BgARP1</Emphasis> enhances salt tolerance in transgenic <Emphasis Type="Italic">Arabidopsis</Emphasis> plants

We have previously reported that expression of salt-responsive genes, including Bruguiera gymnorhiza ankyrin repeat protein 1 (BgARP1), enhances salt tolerance in both Agrobacterium tumefaciens and Arabidopsis. In this report, we further characterized BgARP1-expressing Arabidopsis to elucidate the role of BgARP1 in salt tolerance. BgARP1-expressing plants exhibited more vigorous growth than wild-type plants on MS plates containing 125–175 mM NaCl. Real-time PCR analysis showed enhanced induction of osmotin34 in the 2-week-old transformants under 125 mM NaCl. It was also showed that induction of typical salt-responsive genes, including RD29A, RD29B, and RD22, was blunted and delayed in the 4-week-old transformants during 24 h after 200 mM NaCl treatment. Ion content analysis showed that transgenic plants contained more K⁺, Ca²⁺, and NO₃ ⁻, and less NH₄ ⁺, than wild-type plants grown in 200 mM NaCl. Our results suggest that BgARP1-expressing plants may reduce salt stress by up-regulating osmotin34 gene expression and maintaining K⁺ homeostasis and regulating Ca²⁺ content. These results indicate that BgARP1 is functional on a heterogeneous background.     

The Effect of the "D" Genome from Synthetic Wheat Lines in Anther Culture Responses

Synthetic lines of wheat were compared in order to determine the effect of the “D” genome on embryo induction and plant regeneration from anther culture. The experiments were performed using 6 genotypes of wheat in a randomized block design with 4 replications. In all cases except one, addition of the “D” genome from T. tauschii increased the efficiency of embryo formation and green plantlet production. This positive effect of the “D” genome does not exclude the existence of possible interactions between the three genomes (A, B and D) and/or the effect of each genome separately.     

Genetic analysis of resistance to <Emphasis Type="Italic">soil-borne wheat mosaic virus</Emphasis> derived from <Emphasis Type="Italic">Aegilops tauschii</Emphasis>

Genetic Analysis of Resistance to Soil-Borne Wheat Mosaic Virus Derived from Aegilops tauschii. Euphytica. Soil-Borne Wheat Mosaic Virus (SBWMV), vectored by the soil inhabiting organism Polymyxa graminis, causes damage to wheat (Triticum aestivum) yields in most of the wheat growing regions of the world. In localized fields, the entire crop may be lost to the virus. Although many winter wheat cultivars contain resistance to SBWMV, the inheritance of resistance is poorly understood. A linkage analysis of a segregating recombinant inbred line population from the cross KS96WGRC40 × Wichita identified a gene of major effect conferring resistance to SBWMV in the germplasm KS96WGRC40. The SBWMV resistance gene within KS96WGRC40 was derived from accession TA2397 of Aegilops taushcii and is located on the long arm of chromosome 5D, flanked by microsatellite markers Xcfd10 and Xbarc144. The relationship of this locus with a previously identified QTL for SBWMV resistance and the Sbm1 gene conferring resistance to soil-borne cereal mosaic virus is not known, but suggests that a gene on 5DL conferring resistance to both viruses may be present in T. aestivum, as well as the D-genome donor Ae. tauschii.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated transformation of faba bean <Emphasis Type="Italic">(Vicia faba</Emphasis> L.) using embryo axes

A system for the production of transgenic faba bean by Agrobacterium-mediated transformation was developed. This system is based upon direct shoot organogenesis after transformation of meristematic cells derived from embryo axes. Explants were co-cultivated with A. tumefaciens strain EHA105/pGlsfa, which harbored a binary vector containing a gene encoding a sulphur rich sunflower albumin (SFA8) linked to the bar gene. Strain EHA 101/pAN109 carrying the binary plasmid containing the coding sequence of a mutant aspartate kinase gene (lysC) from E. coli in combination with neomycinphosphotransferase II gene (nptII) was used as well. The coding sequences of SFA8 and LysC genes were fused to seed specific promoters, either Vicia faba legumin B4 promoter (LeB4) or phaseolin promoter, respectively. Seven phosphinothricin (PPT) resistant clones from Mythos and Albatross cultivars were recovered. Integration, inheritance and expression of the transgenes were confirmed by Southern blot, PCR, enzyme activity assay and Western blot.     

A novel frameshift mutant allele, <Emphasis Type="Italic">fzp</Emphasis>-<Emphasis Type="Italic">10</Emphasis>, affecting the panicle architecture of rice

The rice FRIZZY PANICLE (FZP) locus on chromosome 7, in which an ERF and acidic domain are present, is concerned with the regulation of spikelet meristem identity and the determination of panicle architecture. Many fzp mutants drastically alter panicle morphology with higher-order rachis-branches developing successively instead of the development of floral organs in these mutants. A new mutant showing the same fzp phenotype was induced by γ-ray irradiation of seeds of a rice cultivar “Gimbozu”. Examination of this fzp-like mutant for its allelism to a known allele of fzp-1, nucleotide sequence, and panicle and agronomic characteristics clearly indicated that the allele of this fzp-like mutant is located in FZP. Because there is a previously identified allele called fzp-9, we designated this new allele as fzp-10. fzp-10 has a single nucleotide (cytosine) deletion between the ERF domain and the acidic domain, which results in a frameshift mutation and a premature stop codon, thereby altering the C-terminus of the encoded protein. The degree of higher-order branching in the panicles was significantly reduced in the fzp-10 mutant compared with that of fzp-1. Moreover, the fzp-10 mutant showed highly depressed culm and panicle lengths and panicle number, as well as delayed heading dates compared with its wild type and also with fzp-1. fzp-10 has several new characteristics, its altered nucleotide position and severity of phenotype alteration, and, therefore, could be a new gene resource to examine the function of FZP and the determination of rice panicle architecture.     

RDA derived <Emphasis Type="Italic">Oryza minuta</Emphasis>-specific clones to probe genomic conservation across <Emphasis Type="Italic">Oryza</Emphasis> and introgression into rice (<Emphasis Type="Italic">O. sativa</Emphasis> L.)

Molecular markers have been successfully used in rice breeding however available markers based on Oryza sativa sequences are not efficient to monitor alien introgression from distant genomes of Oryza. We developed O. minuta (2n = 48, BBCC)-specific clones comprising of 105 clones (266–715 bp) from the initial library composed of 1,920 clones against O. sativa by representational difference analysis (RDA), a subtractive cloning method and validated through Southern blot hybridization. Chromosomal location of O. minuta-specific clones was identified by hybridization with the genomic DNA of eight monosomic alien additional lines (MAALs). The 37 clones were located either on chromosomes 6, 7, or 12. Different hybridization patterns between O. minuta-specific clones and wild species such as O. punctata, O. officinalis, O. rhizomatis, O. australiensis, and O. ridleyi were observed indicating conservation of the O. minuta fragments across Oryza spp. A highly repetitive clone, OmSC45 hybridized with O. minuta and O. australiensis (EE), and was found in 6,500 and 9,000 copies, respectively, suggesting an independent and exponential amplification of the fragment in both species during the evolution of Oryza. Hybridization of 105 O. minuta specific clones with BB- and CC-genome wild Oryza species resulted in the identification of 4 BB-genome-specific and 14 CC-genome-specific clones. OmSC45 was identified as a fragment of RIRE1, an LTR-retrotransposon. Furthermore this clone was introgressed from O. minuta into the advanced breeding lines of O. sativa.     

Fine mapping of a <Emphasis Type="Italic">Xantha</Emphasis> mutation in rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

The recessive mutation of the XANTHA gene (XNT) transforms seedlings and plants into a yellow color, visually distinguishable from normal (green) rice. Thus, it has been introduced into male sterile lines as a distinct marker for rapidly testing and efficiently increasing varietal purity in seed and paddy production of hybrid rice. To identify closely linked markers and eventually isolate the XNT gene, two mapping populations were developed by crossing the xantha mutant line Huangyu B (indica) with two wild type japonica varieties; a total of 1,720 mutant type F₂ individuals were analyzed for fine mapping using polymorphic InDel markers and high dense microsatellite markers. The XNT gene was mapped on chromosome 11, within in a fragment of ~100 kb, where 13 genes are annotated. The NP_001067671.1 gene within the delimited region is likely to be a candidate XNT gene, since it encodes ATP-dependent chloroplast protease ATP-binding subunit clp A. However, no sequence differences were observed between the mutant and its parent. Bioinformatics analysis demonstrated that four chlorophyll deficient mutations that were previously mapped on the same chromosome are located outside the XNT region, indicating XNT is a new gene. The results provide useful DNA markers not only for marker assisted selection of the xantha trait but also its eventual cloning.     

Ploidy level studies on the <Emphasis Type="Italic">Dioscorea cayenensis</Emphasis>/<Emphasis Type="Italic">Dioscorea rotundata</Emphasis> complex core set

The Guinea yams, Dioscorea cayenensis Lam. and D. rotundata Poir. (D. cayenensis–D. rotundata complex), represent a highly important crop, widely distributed in the humid and semi-humid tropics. The ploidy levels of 170 accessions of the core set of Guinea yams from West African countries was determined using flow cytometry with propidium iodide staining. One hundred and eight of the genotypes were found to be tetraploid, 47 were hexaploid and five were octoploid. One mixoploid individual containing tetraploid and hexaploid nuclei was also detected. A deeper analysis considering each separate taxon revealed that while for D. rotundata the majority of individuals were tetraploid, for D. cayenensis this ploidy level was not detected in any of the accessions. Also, no association between ploidy level and place of cultivation was found for the evaluated germplasm. The obtained data is highly valuable for breeding programs of Guinea yam, especially for the optimization of future hybridization experiments directed to the genetic improvement of this economically important crop.     

Morphological,cytological and reproductive characterization of tri-species hybrids (GOS) between <Emphasis Type="Italic">Pennisetum glaucum</Emphasis>, <Emphasis Type="Italic">P. orientale</Emphasis> and <Emphasis Type="Italic">P. squamulatum</Emphasis>

New tri-species hybrids (GOS) in the genus Pennisetum involving the cultivated species pearl millet (P. glaucum L.) and two wild species, viz. P. squamulatum Fresen and P. orientale L. C. Rich, are reported. Six hybrid plants were recovered after crossing a backcross hybrid (2n = 3x = 23, GGO) between P. glaucum (2n = 2x = 14, GG) and P. orientale (2n = 2x = 18, OO) with F₁s (2n = 6x = 42, GGSSSS) between P. glaucum (2n = 4x = 28, GGGG) and P. squamulatum (2n = 8x = 56, SSSSSSSS). The hybrids were perennial, morphologically intermediate to their parents, and represented characters from the three contributing species. The hybrids contained 2n = 44 chromosomes (GGGSSO) representing 21, 14 and nine chromosomes from P. glaucum, P. squamulatum and P. orientale, respectively. Meiotic and flow-cytometric analysis suggested origin of these hybrids from unreduced female and reduced male gametes. Average chromosome configuration (8.42_I + 14.32_II + 1.62_III + 0.52_IV) at Meiosis showed limited inter-genomic pairing indicating absence of significant homology between the three genomes. The hybrids were male sterile (except one) and highly aposporous. P. orientale was identified to induce apospory in hybrid background with P. glaucum at diploid and above levels, though it was quantitatively affected by genomic doses from sexual parent. A case of inducible and recurrent apospory is presented whereby a transition from Polygonum-type sexual embryo-sacs to Panicum-type aposporous embryo-sacs was observed in diploid interspecific hybrids. Results supported independent origin and partitioning of the three apomixis-components (apomeiosis, parthenogenesis, and functional endosperm development), reported for the first time in Pennisetum. Potential utilization of GOS hybrids in understanding genome interactions involved in complex traits, such as perenniality and apomixis, is discussed.     

Production of interspecific hybrids between <Emphasis Type="Italic">Lilium longiflorum</Emphasis> and <Emphasis Type="Italic">L. lophophorum</Emphasis> var. <Emphasis Type="Italic">linearifolium</Emphasis> via ovule culture at early stage

Interspecific hybridization was carried out between Lilium longiflorum and L. lophophorum var. linearifolium by using the cut style method of pollination, as a contrast, intraspecific hybridization between L. longiflorum ‘Gelria’ and L. longiflorum was also made, but no mature seeds and offspring were obtained from the two combinations under in vivo condition. Ovules excised from each carpel 5–35 days after pollination (DAP) were cultured on B5 or half-strength B5 medium containing sucrose at different concentrations in vitro. In L. longiflorum × L. lophophorum var. linearifolium, only 1.17% of ovules excised at 10 DAP developed into seedlings, and in L. longiflorum ‘Gelria’ × L. longiflorum, only 0.99% of ovules excised at 25 DAP developed into seedlings; none of the ovules excised at other different DAP in the two cross combinations produced any seedlings. The results showed that interspecific hybridization had a more serious post-fertilization barrier than the intraspecific hybridization, and that a lower concentration (3%) of sucrose led to better embryo development and higher percentage of seedlings in ovule cultures. All hybrid seedlings obtained were successfully transplanted to soil and grew normally. The progenies investigated were identified as true hybrids based on inter-simple sequence repeat (ISSR) analysis.     

Identification and mapping of quantitative resistance to late blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) in <Emphasis Type="Italic">Solanum habrochaites</Emphasis> LA1777

Late blight (Phytophthora infestans) can have devastating effects on tomato production over the whole world. Most of the commercial cultivars of tomato, Solanum lycopersicum, are susceptible. Qualitative and quantitative resistance has been described in wild relatives of tomato. In general qualitative resistance can more easily be overcome by newly evolved isolates. Screening of three S. habrochaites accessions (LA1033, LA2099 and LA1777) through a whole plant assay showed that accession LA1777 had a good level of resistance to several isolates of P. infestans. To explore the potential in this wild species, an introgression line (IL) population of S. habrochaites LA1777 was used to screen individual chromosome regions of the wild species by a detached leaf assay. Two major isolates (T_1,2 and T_1,2,4) were used and two parameters were measured: lesion size (LS), and disease incidence (DI). Substantial variation was observed between the individual lines. QTLs were identified for LS but not for DI. The presence of five QTLs derived from LA1777 (Rlbq4a, Rlbq4b, Rlbq7, Rlbq8 and Rlbq12) results in unambiguous higher levels of resistance. All QTLs co-localized with previously described QTLs from S. habrochaites LA2099 except QTL Rlbq4b, which is therefore a novel QTL.