Response to selection for improved nitrogen fixation in common bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Breeding for high symbiotic nitrogen (N) fixation (SNF) in common bean (Phaseolus vulgaris L.) is expected to contribute to reduced application of chemical fertilizers in cropping systems involving common bean. The magnitude of variation and the genetic and phenotypic correlation among seed yield, SNF, estimated as the percentage of nitrogen derived from atmosphere, and related traits were studied in a population of 140 F₄-derived F₅ recombinant inbred lines, developed from a cross between low- and high-SNF bean genotypes ‘Sanilac’ and ‘Mist’, respectively. The experiment was conducted in a total of five location-years in Ontario, Canada, from 2011 to 2013. These location-years were grouped into stress- and optimum moisture test sites, based on the total precipitation during the growing season. In each test site two nitrogen supply management strategies, SNF-dependent and N fertilizer-dependent, were simulated separately in the field by inoculating the seed with a commercial Rhizobium leguminosarum bv. phaseoli and by application of N fertilizers at 100 kg ha^?1, respectively. The genetic variation was significant for seed yield, SNF and related traits. The heritability of the traits ranged from 14 to 71% and 4 to 25% in optimum moisture and in stress environments, respectively. No significant correlation between SNF and seed yield indicated that selection for high SNF does not necessarily lead to greater seed yield and that selection for both traits should be performed simultaneously.     

Fingerprinting temperate <Emphasis Type="Italic">japonica</Emphasis> and tropical <Emphasis Type="Italic">indica</Emphasis> rice genotypes by comparative analysis of DNA markers

This paper describes the relative efficiency of three marker systems, RAPD, ISSR, and AFLP, in terms of fingerprinting 14 rice genotypes consisting of seven temperatejaponica rice cultivars, three indica near-isogenic lines, three indica introgression lines, and one breeding line of japonica type adapted to high-altitude areas of the tropics with cold tolerance genes. Fourteen RAPD, 21 ISSR, and 8 AFLP primers could produce 970 loci, with the highest average number of loci (92.5) generated by AFLP. Although polymorphic bands in the genotypes were detected by all marker assays, the AFLP assay discriminated the genotypes effectively with a robust discriminating power (0.99), followed by ISSR (0.76) and RAPD (0.61). While significant polymorphism was detected among the genotypes of japonica and indica through analysis of molecular variance (AMOVA), relatively low polymorphism was detected within the genotypes of japonica rice cultivars. The correlation coefficients of similarity were significant for the three marker systems used, but only the AFLP assay effectively differentiated all tested rice lines. Fingerprinting of backcross-derived resistant progenies using ISSR and AFLP markers easily detected progenies having a maximum rate of recovery for the recurrent parent genome and suggested that our fingerprinting approach adopting the ‘undefined-element-amplifying’ DNA marker system is suitable for incorporating useful alleles from the indica donor genome into the genome of temperate japonica rice cultivars with the least impact of deleterious linkage drag.     

Mapping QTL for seed yield and canning quality following processing of black bean (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Quantitative trait loci (QTL) analysis was conducted to identify QTL for seed yield and color retention following processing of a recombinant inbred line (RIL) black bean population. A population of 96 RILs were derived from the cross of black bean cultivars ‘Jaguar’ and 115M and evaluated in replicated trials at one location over 4 years (2004–2007) in Michigan. A 119-point genetic map constructed using simple sequence repeat (SSR), sequence related amplified polymorphism (SRAP), target region amplified polymorphism (TRAP) and phenotypic markers spanned fifteen linkage groups (LG) or 460 cM of the bean genome. Fourteen QTL for yield and color retention in four environments were identified by composite interval mapping on six linkage groups. A major QTL SY10.2^J115 for seed yield was identified on LG B10 with additional QTL on B3, B5, and B11. Color retention following processing was associated with loci on B1, B3, B5, B8, and B11. 115M possessed positive alleles for yield, but negative alleles for color retention. Some QTL for yield and color retention co-localized with regions identified in previous studies while others, particularly for color retention, were unique. Additional QTL for agronomic and canning quality traits were detected and individual contributions to future black bean breeding are discussed.     

Dominant gene for common bean resistance to common bacterial blight caused by <Emphasis Type="Italic">Xanthomonas</Emphasis><Emphasis Type="Italic">axonopodis</Emphasis> pv. <Emphasis Type="Italic">phaseoli</Emphasis>

The common bacterial blight pathogen [Xanthomonas axonopodis pv. phaseoli (Xap)] is a limiting factor for common bean (Phaseolus vulgaris L.) production worldwide and resistance to the pathogen in most commercial cultivars is inadequate. Variability in virulence of the bacterial pathogen has been observed in strains isolated from Puerto Rico and Central America. A few common bean lines show a differential reaction when inoculated with different Xap strains, indicating the presence of pathogenic races. In order to study the inheritance of resistance to common bacterial blight in common bean, a breeding line that showed a differential foliar reaction to Xap strains was selected and was crossed with a susceptible parent. The inheritance of resistance to one of the selected Xap races was determined by analysis of segregation patterns in the F₁, F₂, F₃ and F₄ generations from the cross between the resistant parent PR0313-58 and the susceptible parent ‘Rosada Nativa’. The F₁, F₂ and F₃ generations were tested under greenhouse conditions. Resistant and susceptible F_3:4 sister lines were tested in the field. The statistical analysis of all generations followed the model for a dominant resistance gene. The resistant phenotype was found to co-segregate with the SCAR SAP6 marker, located on LG 10. These results fit the hypothesis that resistance is controlled by a single dominant gene. The symbol proposed for the resistance gene is Xap-1 and for the bacterial race, XapV1.     

Breeding for salinity resistance in mulberry (<Emphasis Type="Italic">Morus</Emphasis> spp.)

Late maturity α-amylase (LMA) is a genetic defect that is fairly widely spread in bread wheat (Triticum aestivum L.) germplasm, and recently detected in durum cultivars, which can result in unacceptably high α-amylase activity (low falling number) in ripe grain. LMA has also been observed at unexpectedly high frequency and severity in synthetic hexaploid wheats derived from the interspecific hybridisation of Triticum durum (AABB) and Aegilops tauschii (DD). Since synthetic hexaploids represent an important new source of resistances/tolerances to a range of biotic and abiotic stresses for wheat breeders, there is a pressing need to understand the mechanisms involved in LMA in synthetics and develop strategies for avoiding its adverse effects on grain quality. The objectives of this study were to firstly, compare the LMA phenotype of synthetics that varied for plant height, secondly, to characterise the LMA phenotype in groups of synthetics derived from the same durum parents and finally to determine whether LMA in primary synthetics is associated with the QTL previously reported in conventional bread wheat. More than 250 synthetic hexaploids, a range of durum cultivars and a doubled haploid population derived from Worrakatta (non-LMA) × AUS29663 (high LMA synthetic) were phenotyped and genotyped with markers reported to be linked to LMA in conventional bread wheat and markers diagnostic for the semi-dwarfing gene, Rht1. More than 85% of synthetics were prone to LMA, approximately 60% ranked as very high. Genetic control of LMA in synthetic hexaploids appeared to involve QTL located on 7B, and to a lesser extent 3B, similar to bread wheats. However, the LMA phenotype of many synthetic hexaploids appeared to be more extreme than could be explained by comparisons with bread wheat even taking into account the apparent absence of Rht1 in most genotypes. Other mechanisms, possibly triggered by the interaction between the AABB and DD genomes cannot be excluded. The presence of wild type rht1 in most synthetic hexaploids and their extreme height is difficult to reconcile with the semi-dwarf, Rht1, stature of many of the durums used in the interspecific hybridisation process. Mechanisms that could explain this observation remain unclear.     

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.     

<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.     

Increased resistance to <Emphasis Type="Italic">Ustilago zeae</Emphasis> and <Emphasis Type="Italic">Fusarium verticilliodes</Emphasis> in maize inbred lines bred for <Emphasis Type="Italic">Fusarium graminearum</Emphasis> resistance

Preliminary field observations in our maize breeding nurseries indicated that breeding for improved resistance to gibberella ear rot (Fusarium graminearum) in maize may indirectly select for resistance to another ear disease, common smut (Ustilago zeae). To investigate this, we compared the disease severity ratings obtained on 189 maize inbreds, eight of which included our inbreds developed with selection for gibberella ear rot resistance after field inoculation and breeding for 8–10 years. No correlation was found between disease severities for the 189 inbreds but the eight gibberella-resistant lines were consistently more resistant to smut. To further examine this relationship and to determine if these eight inbreds would be useful for developing inbreds with either common smut or fusarium ear rot (F. verticilliodes) resistance, we conducted a Griffing’s diallel analysis on six inbreds of maize, four with high levels of gibberella ear rot resistance representing all of the pedigree groups in our eight gibberella lines, and two with very low levels. Our most gibberella ear rot resistant inbreds, CO433 and CO441, had the lowest disease ratings for all three diseases, the consistently largest general combining ability effects and several significant specific combining ability effects. It was concluded that some inbreds bred specifically for gibberella ear rot would also be useful in breeding for resistance to common smut and fusarium ear rot.     

Variation of <Emphasis Type="Italic">Wx</Emphasis> microsatellite allele,waxy allele distribution and differentiation of chloroplast DNA in a collection of Thai rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Eighty-two varieties of rice from different regions in Thailand were selected to explore the Waxy (Wx)gene diversity and indica-japonica differentiation of chloroplast DNA. A comparison of the 5 splice site in the first intron was made between glutinous and nonglutinous rice. It revealed that non-glutinous with low-amylose content and glutinous rice were characterized as the Wx^b allele based on the G-to-T base substitution, whereas non-glutinous rice with intermediate and high amylose carried the Wx^a allele. Four Wx microsatellite alleles, (CT)_n repeat, (n = 16,17,18 and 19) were found in glutinous rice. In contrast, non-glutinous rice showed five Wx microsatellite alleles (n = 11, 16, 17, 18 and 19). The (CT)₁₇ allele was prominent allele in Thai population, while the (CT)₁₁ allele was found only in intermediate and high amylose rice varieties from southern Thailand. Almost all of upland rice grown by various ethnic groups in northern Thailand were characterized as japonica type based on their having the PstI-12 fragment in their cpDNA, whereas most of rainfed lowland varieties from other regions of Thailand were indica. This exploration of DNA-based genetic markers is important, as it enhances our ability to describe and manipulate sources of genetic variation for rice breeding programs.     

Successful induction of trigenomic hexaploid <Emphasis Type="Italic">Brassica</Emphasis> from a triploid hybrid of <Emphasis Type="Italic">B.</Emphasis><Emphasis Type="Italic">napus</Emphasis> L. and <Emphasis Type="Italic">B. nigra</Emphasis> (L.) Koch

A triploid hybrid with an ABC genome constitution, produced from an interspecific cross between Brassica napus (AACC genome) and B. nigra (BB genome), was used as source material for chromosome doubling. Two approaches were undertaken for the production of hexaploids: firstly, by self-pollination and open-pollination of the triploid hybrid; and secondly, by application of colchicine to axillary meristems of triploid plants. Sixteen seeds were harvested from triploid plants and two seedlings were confirmed to be hexaploids with 54 chromosomes. Pollen viability increased from 13% in triploids to a maximum of 49% in hexaploids. Petal length increased from 1.3 cm (triploid) to 1.9 cm and 1.8 cm in the two hexaploids and longest stamen length increased from 0.9 cm (triploid) to 1.1 cm in the hexaploids. Pollen grains were longer in hexaploids (43.7 and 46.3 μm) compared to the triploid (25.4 μm). A few aneuploid offsprings were also observed, with chromosome number ranging from 34 to 48. This study shows that trigenomic hexaploids can be produced in Brassica through interspecific hybridisation of B. napus and B. nigra followed by colchicine treatment.     

A high-density integrative linkage map for <Emphasis Type="Italic">Gossypium hirsutum</Emphasis>

Sequence-related amplified polymorphism (SRAP) combined with SSRs, RAPDs, and RGAPs was used to construct a high density genetic map for a F₂ population derived from the cross DH962 (G. hirsutum accession) × Jimian5 (G. hirsutum cultivar). A total of 4,096 SRAP primer combinations, 6310 SSRs, 600 RAPDs, and 10 RGAPs produced 331, 156, 17 and 2 polymorphic loci, respectively. Among the 506 loci obtained, 471 loci (309 SRAPs, 144 SSRs, 16 RAPDs and 2 RGAPs) were assigned to 51 linkage groups. Of these, 29 linkage groups were assigned to corresponding chromosomes by SSR markers with known chromosome locations. The map covered 3070.2 cM with a mean density of 6.5 cM per locus. The segregation distortion in this population was 9.49%, and these distorted loci tend to cluster at the end of linkage groups or in minor clusters on linkage groups. The majority of SRAPs in this map provided an effective tool for map construction in G. hirsutum despite of its low polymorphism. This high-density linkage map will be useful for further genetic studies in Upland cotton, including mapping of loci controlling quantitative traits, and comparative and integrative analysis with other interspecific and intraspecific linkage maps in cotton.     

Developing and validating microsatellite markers in elephant grass (<Emphasis Type="Italic">Pennisetum purpureum</Emphasis> S.)

Elephant grass [Pennisetum purpureum S.; syn. Cenchrus purpureus (Schumach.) Morrone] is an important global forage crop and is recognized for high yields of herbage with good nutritive value. It also has high biomass potential to be utilized as a biofuel feedstock. Whereas several previous genetic studies adapted simple sequence repeat (SSR) markers from pearl millet [Pennisetum glaucum (L.) R.Br.] for investigations in elephant grass, the present study developed SSR markers from 3536 DNA sequences derived from 16 elephant grass entries. A total of 3866 SSRs were identified including 1028 monomeric, 2019 dimeric, 735 trimeric, 49 tetrameric, 20 pentameric and 15 hexameric repeat motifs. Three hundred and seven sequences contained more than one repeated motif, and 154 SSRs were present in compound formation. Susequenctly,  four elephant grass and two pearl millet genotypes were chosen to validate 727 SSR markers. Of these, 628 markers produced visually detectable amplification products, including 73 (11.6%) polymorphic ones across all six genotypes. Polymorphism between the four elephant grass genotypes was revealed by 316 (50.6%) markers with diversity index values ranging from 0.75 to 0.38. Dimeric SSRs had the highest polymorphic rate (48.7%). These validated SSR markers had 58.6% (368 of 628) transferability rate to pearl millet. The availability of these polymorphic SSR markers will support advanced genetic studies in P. purpureum and its relatives.     

Genetic effects of introgression genomic components from Sea Island cotton (<Emphasis Type="Italic">Gossypium barbadense</Emphasis> L.) on fiber related traits in upland cotton (<Emphasis Type="Italic">G</Emphasis>. <Emphasis Type="Italic">hirsutum</Emphasis> L.)

The germplasm with exotic genomic components especially from Sea Island cotton (Gossypium barbadense L. Gb) is the dominant genetic resources to enhance fiber quality of upland cotton (G. hirsutum L., Gh). Due to low efficiency of phenotypic evaluation and selection on fiber quality, genetic dissection of favorable alleles using molecular markers is essential. Genetic dissection on putative Gb introgressions related to fiber traits were conducted by SSR markers with mapping populations derived from a cross between Luyuan343 (LY343), a superior fiber quality introgression line (IL) with genomic components from Gb, and an elite Upland cotton cv. Lumianyan#22 (LMY22). Among 82 polymorphic loci screened out from 4050 SSRs, 42 were identified as putative introgression alleles. A total of 29 fiber-related QTLs (23 for fiber quality and six for lint percentage) were detected and most of which clustered on the putative Gb introgression chromosomal segments of Chr.2, Chr.16, Chr.23 and Chr.25. As expected, a majority of favorable alleles of fiber quality QTLs (12/17, not considering the QTLs for fiber fineness) came from the IL parent and most of which (11/12) were conferred by the introgression genomic components while three of the six (3/6) favorable alleles for lint percentage came from the Gh parent. Validation of these QTLs using an F₈ breeding population from the same cross made previously indicated that 13 out of 29 QTLs showed considerable stability. The results suggest that fiber quality improvement using the introgression components could be facilitated by marker-assisted selection in cotton breeding program.     

A linkage map for flowering dogwood (<Emphasis Type="Italic">Cornus florida</Emphasis> L.) based on microsatellite markers

A genetic linkage map of flowering dogwood (Cornus florida L.) was constructed using 94 individuals derived from a cross of two F₁ trees designated 97-6 and 97-7, which were originally from a cross between ‘Appalachian Spring’ and ‘Cherokee Brave’. Out of approximately 800 SSR loci examined, 271 were polymorphic between ‘Appalachian Spring’ and ‘Cherokee Brave’, but were monomorphic between 97-6 and 97-7. These 271 segregating markers were used to build a linkage map for flowering dogwood. Eleven linkage groups were obtained with a log-of-odds (LOD) value of 6.0 using JoinMap® 4.0 software, which matches the chromosome number of flowering dogwood haploid genome. This linkage map consisted of 255 SSR loci, spanned a total of 1,175 centimorgans (cM) with an average internal distance of 4.6 cM. Several larger gaps and slight clustering of markers were present on this linkage map. This is the first linkage map of flowering dogwood and will be a fundamental tool for new gene identification and marker-assisted selection in our flowering dogwood breeding program.     

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.     

Sensory changes related to breeding for plant architecture and resistance to viruses and anthracnose in bean market class Fabada (<Emphasis Type="Italic">Phaseolus vulgaris</Emphasis> L.)

Consumers hold landraces in high esteem and often consider that breeding programs lead to a loss of sensory quality, although consumers’ opinions have not been scientifically confirmed. As a model case of study we recorded seed sensory traits in six inbred common bean lines classified in the market class Fabada obtained by backcrossing and/or pedigree selection (to change the plant architecture and increase resistances) and then cultivated in two environments in two consecutive years. A sensory panel noted differences among all the inbreds, but only the introduction of the gene fin (determinate growth habit) seemed to be associated to a loss of quality (rougher seed coat and lower percentage of whole beans after cooking). As theoretically expected, changes in sensory traits were smaller in inbreds obtained through simple backcrossing than in those obtained through pedigree selection after inbred crossing. Large differences in sensory traits due to environmental factors were also recorded (similar to the genetic effects in magnitude), especially soil type and rainfall around flowering. These results make it highly recommendable to evaluate sensory attributes in the course of breeding programs to ensure that the sensory quality is not lost. Given the importance of the environmental component in the phenotypic variation of the sensory traits studied, crop location and management also appear as key factors for the obtainment of optimum products.     

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.     

Biochemical markers for cabbage seedpod weevil (<Emphasis Type="Italic">Ceutorhynchus obstrictus</Emphasis> (Marsham)) resistance in canola (<Emphasis Type="Italic">Brassica napus</Emphasis> L.)

In the last decade, the cabbage seedpod weevil (Ceutorhynchus obstrictus (Marsham)) has become a major insect pest of canola (Brassica napus L.) in Canada reducing seed yields up to 35%. Therefore, the benefits of developing weevil resistant germplasm to canola breeders and the industry would reduce input costs, pesticide use, environmental degradation and increase yield. Yellow mustard (Sinapis alba L.) is resistant to C. obstrictus (CSPW), although the exact mechanism is not known (McCaffrey et al. 1999). A unique canola population was generated at the University of Guelph from a cross between B. napus and S. alba through embryo rescue and backcrossed to canola several times prior to double haploid (DH) production. Approximately one-half of this DH population had canola quality glucosinolate concentration (<16 μmol/g) and was used for further breeding. The hypothesis was that some DH progeny from this cross inherited resistance to CSPW from S. alba. Weevil infestation levels were assessed for the B. napus × S. alba BC2 and BC3 DH populations in the field over 7 years in Alberta where weevil pressure is strong to establish the resistant or susceptible status of these lines. The basic objectives for this study were to confirm field resistance in the B. napus × S. alba germplasm in Ontario and to identify any biochemical markers associated with resistance/susceptibility. Canola doubled haploid lines derived from BC2 or BC3 families were field screened for resistance (R) followed by chemical analysis of glucosinolates to detect biochemical polymorphisms correlated with CSPW resistance using High Performance Liquid Chromatography (HPLC). Two polymorphic peaks were found, one each, from extracts of upper cauline leaves and Stage 3 pod seed, with retention times of ~23 and 19 min, respectively. These HPLC peaks consistently correlated with larval infestation data and the peak differences between R and S DH lines were significant. Therefore, these two peaks can be considered as biochemical markers in this breeding germplasm and may play a role in rapid and early detection of CSPW resistance.     

A simple,rapid and reliable bioassay for evaluating seedling vigor under submergence in <Emphasis Type="Italic">indica</Emphasis> and <Emphasis Type="Italic">japonica</Emphasis> rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Submergence is a major stress causing yield losses particularly in the direct-seeded rice cultivation system and necessitates the development of a simple, rapid and reliable bioassay for a large scale screening of rice germplasms with tolerance against submergence stress. We developed two new bioassay methods that were based primarily on the seedling vigor evaluated by the ability of fast shoot elongation under submerged conditions, and compared their effectiveness with two other available methods. All four bioassay methods using cultivars of 7 indica and 6 japonica types revealed significant and consistent cultivar differences in seedling vigor under submergence and/or submergence tolerance. Japonica cultivars were more vigorous than indica cultivars, with Nipponbare being the most vigorous. The simplest test tube method showed the highest correlations to all other methods. Our results suggest that seedling vigor serves as a submergence avoidance mechanism and confers tolerance on rice seedlings to flooding during early crop establishment. A possible relationship is discussed between seedling vigor based on fast shoot elongation and submergence tolerance defined by recovery from submergence stress.     

Genetic analysis of <Emphasis Type="Italic">Jatropha</Emphasis> species and interspecific hybrids of <Emphasis Type="Italic">Jatropha curcas</Emphasis> using nuclear and organelle specific markers

The present study aims at characterization of Jatropha species occurring in India using nuclear and organelle specific primers for supporting interspecific gene transfer. DNA from 34 accessions comprising eight agronomically important species (Jatropha curcas, J. gossypifolia, J. glandulifera, J. integerrima, J. podagrica, J. multifida, J. villosa, J. villosa. var. ramnadensis, J. maheshwarii) and a natural hybrid, J. tanjorensis were subjected to molecular analysis using 200 RAPD, 100 ISSR and 50 organelle specific microsatellite primers from other angiosperms. The nuclear marker systems revealed high interspecific genetic variation (98.5% polymorphism) corroborating with the morphological differentiation of the species used in the study. Ten organelle specific microsatellite primers resulted in single, discrete bands of which three were functional disclosing polymorphism among Jatropha species. The PCR products obtained with organelle specific primers were subjected to sequence analysis. PCR products from two consensus chloroplast microsatellite primer pairs (ccmp6 and 10) revealed variable number of T and A residues in the intergenic regions of ORF 77–ORF 82 and rp12–rps19 regions, respectively in Jatropha. Artificial hybrids were produced between J. curcas and all Jatropha species used in the study with the exception of J. podagrica. Characterization of F₁ hybrids using polymorphic primers specific to the respective parental species confirmed the hybridity of the interspecific hybrids. Characterization of both natural and artificially produced hybrids using chloroplast specific markers revealed maternal inheritance of the markers. While the RAPD and ISSR markers confirmed J. tanjorensis as a natural hybrid between J. gossypifolia and J. curcas, the ccmp primers (ccmp6 and 10) unequivocally established J. gossypifolia as the maternal parent. Evaluation of backcross interspecific derivatives of cross involving J. curcas and J. integerrima indicate scope for prebreeding and genetic enhancement of Jatropha curcas through interspecific hybridization.