Resistance screening of breeding lines and commercial tomato cultivars for <Emphasis Type="Italic">Meloidogyne incognita</Emphasis> and <Emphasis Type="Italic">M. javanica</Emphasis> populations (Nematoda) from Ethiopia

Soil and root samples were collected from major tomato growing areas of Ethiopia during the 2012/2013 growing season to identify root-knot nematode problems. DNA-based and isozyme techniques revealed that Meloidogyne incognita and M. javanica were the predominant Meloidogyne species across the sampled areas. The aggressiveness of different populations of these species was assessed on tomato cultivars Marmande and Moneymaker. The two most aggressive populations of each species were selected and further tested on 33 tomato genotypes. The resistance screening and mechanism of resistance was performed after inoculation with 100 freshly hatched (<24 h) second-stage juveniles (J2). Eight weeks after inoculation the number of egg masses produced on each cultivar was assessed. For the resistance mechanism study, J2 penetration and their subsequent development inside the tomato roots were examined at 1, 2, 4 and 6 weeks after inoculation. On both cultivars Marmande and Moneymaker all M. incognita and M. javanica populations formed a high number of egg masses indicating highly aggressive behaviour. Populations from ‘Jittu’ and ‘Babile’ for M. incognita and ‘Jittu’ and ‘Koka’ for M. javanica were selected as most aggressive. None of the 33 tomato genotypes were immune for these M. incognita and M. javanica populations. However, several tomato genotypes were found to have a significant effect on the number of egg masses produced indicating possible resistance. For M. javanica populations there were more plants from cultivars or breeding lines on which no egg masses were found compared to M. incognita populations. The lowest number of egg masses for both populations of M. incognita was produced on cultivars Bridget40, Galilea, and Irma while for M. javanica it was on Assila, Eden, Galilea, Tisey, CLN-2366A, CLN-2366B and CLN-2366C. Tomato genotypes, time (weeks after inoculation) and their interaction were significant sources of variation for J2 penetration and their subsequent development inside the tomato roots. Differential penetration was found in breeding lines such as CLN-2366A, CLN-2366B and CLN-2366C, but many of the selected tomato genotypes resistance for the tested M. incognita and M. javanica populations were expressed by delayed nematode development. Therefore, developing a simple screening technique to be used by local farmers or extension workers is crucial to facilitate selection of a suitable cultivar.     

Genetic relationships among resynthesized,semi-resynthesized and natural <Emphasis Type="Italic">Brassica napus</Emphasis> L. genotypes

The allopolyploidization event that created cultivated oilseed rape Brassica napus L, followed by intense breeding, reduced its genetic diversity. Resynthesized (RS) B. napus L. obtained by interspecific hybridization between genotypes of B. rapa L. and B. oleracea L. can be a valuable source for broadening genetic diversity in cultivated oilseed rape. In this study, we determined the extent of DNA polymorphism among natural accessions of oilseed rape, resynthesized B. napus, their parental species and double-low quality semi-RS lines carrying the Rfo gene. Using 10 selected primer combinations, 522 polymorphic AFLP markers were scored in the complete set of 100 Brassica sp. To detect relationships between these genotypes, a cluster analysis was performed using the Jaccard’s distance. Resynthesized allopolyploids clustered directly between their diploid parents. Cultivated accessions of oilseed rape created a compact group away from resynthesized allopolyploids as well as semi-RS lines. The natural oilseed rape group, which consists of 49 cultivars and breeding lines of oilseed rape, is characterized by lower genetic diversity than the group of 33 accessions of resynthesized oilseed rape, and the analysis showed that the double-low quality semi-RS lines represent a specific genetic variation of B. napus. The de novo resynthesized B. napus lines and the semi-RS lines of double-low quality generated from them, provide a significant opportunity for enrichment the gene pool of oilseed rape.     

Association mapping reveals loci associated with multiple traits that affect grain yield and adaptation in soft winter wheat

Genome-wide association studies (GWAS) are useful to facilitate crop improvement via enhanced knowledge of marker-trait associations (MTA). A GWAS for grain yield (GY), yield components, and agronomic traits was conducted using a diverse panel of 239 soft red winter wheat (Triticum aestivum) genotypes evaluated across two growing seasons and eight site-years. Analysis of variance showed significant environment, genotype, and genotype-by-environment effects for GY and yield components. Narrow sense heritability of GY (h ²  = 0.48) was moderate compared to other traits including plant height (h ²  = 0.81) and kernel weight (h ²  = 0.77). There were 112 significant MTA (p < 0.0005) detected for eight measured traits using compressed mixed linear models and 5715 single nucleotide polymorphism markers. MTA for GY and agronomic traits coincided with previously reported QTL for winter and spring wheat. Highly significant MTA for GY showed an overall negative allelic effect for the minor allele, indicating selection against these alleles by breeders. Markers associated with multiple traits observed on chromosomes 1A, 2D, 3B, and 4B with positive minor effects serve as potential targets for marker assisted breeding to select for improvement of GY and related traits. Following marker validation, these multi-trait loci have the potential to be utilized for MAS to improve GY and adaptation of soft red winter wheat.     

Induced expression of selected plant defence related genes in pot azalea, <Emphasis Type="Italic">Rhododendron simsii</Emphasis> hybrid

A set of putative marker genes to study plant defense responses against Polyphagotarsonemus latus, a key pest in the production of Rhododendron simsii hybrids, was selected and validated. Genes belonged to the biosynthetic pathway of phytohormones jasmonic acid (JA) (RsLOX, RsAOS, RsAOC, RsOPR3 and RsJMT) and salicylic acid (SA) (RsPAL and RsICS). Furthermore, RsPPO, a putative marker gene for oxidative stress response was successfully cloned from R. simsii. A CTAB-based extraction protocol was optimized to assure excellent RNA quality for subsequent RT-qPCR analysis. The RT-qPCR protocol was extensively tested and RsRG7 and RsRG14 were selected as reference genes from a geNorm pilot study. Validation of the marker genes was done after application with elicitors [methyl jasmonate (MeJA), coronatine, β-aminobutyric acid and acibenzolar-Smethyl] or wounding. Both 100 μM MeJA and 0.1 μM coronatine had a significant effect on the expression of all marker genes. Foliar application of MeJA on the shoots resulted in a significantly earlier response when compared to root application and subsequent sampling of the shoots. Expression patterns after MeJA treatment were generally the same in six R. simsii genotypes: ‘Nordlicht’, ‘Elien’, ‘Aiko Pink’, ‘Michelle Marie’, ‘Mevrouw Gerard Kint’ and ‘Sachsenstern’. Wounding resulted in the same expression patterns as MeJA treatment except for RsJMT. None of the genotypes showed a significant induction of the latter gene 6 h upon wounding. Findings of these experiments indicated that the tolerant genotype ‘Elien’ has low basal expression levels of RsPPO. This might be the first step towards the breeding of mite-tolerant genotypes.     

Genome-wide detection of genetic loci associated with soybean aphid resistance in soybean germplasm PI 603712

Soybean aphid (Aphis glycines Matsumura) has become one of the major pests of soybean [Glycine max (L.) Merr.] in North America since 2000. At least four biotypes of soybean aphid have been confirmed in the United States. Genetic characterization of new sources of soybean aphid resistance will facilitate the expansion of soybean gene pool for soybean aphid resistance and thus will help to develop soybean aphid resistant cultivars. To characterize the genetic basis of soybean aphid resistance in PI 603712, a newly identified resistant germplasm line, 142 F₂ plants derived from the cross ‘Roberts’ × PI 603712 and their parents were evaluated for soybean aphid resistance in the greenhouse, and were genotyped with BARCSoySNP6K Illumina Infinium II BeadChip. A genome-wide molecular linkage map was constructed with 1495 polymorphic SNP markers. QTL analysis revealed that PI 603712 possessed two major loci associated with soybean aphid resistance, located on chromosome 7 and 16, respectively. The locus on chromosome 7 was dominantly expressed and positioned about one Mega-base-pair distant from the previously identified resistance locus Rag1. The locus on chromosome 16 was positioned near the previously identified resistance locus Rag3 and expressed partially dominance or additive effect. Interestingly, two minor loci were also detected on chromosomes 13 and 17 but the alleles from PI 603712 decreased the resistance. In developing soybean aphid resistant cultivars through marker-assisted selection, an appropriate combination of resistance loci should be selected when PI 603712 is used as a donor parent of resistance.     

Characterization of resistance to the wheat stem sawfly in spring wheat landrace accessions from targeted geographic regions of the world

Plant landraces have long been recognized as potential gene pools for biotic and abiotic stress-related genes. This research used spring wheat landrace accessions to identify new sources of resistance to the wheat stem sawfly (WSS) (Cephus cinctus Norton), an important insect pest of wheat in the northern Great Plains of North America. Screening efforts targeted 1409 accessions from six geographical areas of the world where other species of grain sawflies are endemic or where a high frequency of accessions possesses the resistance characteristic of solid stems. Resistance was observed in approximately 14% of accessions. Half of the lines displayed both antixenosis and antibiosis types of resistance. Among the resistant accessions, 41% had solid or semi-solid stems. Molecular genetic screening for haplotypes at the solid stem QTL, Qss.msub.3BL, showed that 15% of lines shared the haplotype derived from ‘S-615’, the original donor of the solid stem trait to North American germplasm. Other haplotypes associated with solid stems were also observed. Haplotype diversity was greater in the center of origin of wheat. Evaluation of a representative set of resistant landrace accessions in replicated field trials at four locations over a three year period identified accessions with potential genes for reduced WSS infestation, increased WSS mortality, and increased indirect defense via parasitoids. Exploitation of distinct types of plant defense will expand the genetic diversity for WSS resistance currently present in elite breeding lines.     

Characterization of common bean wild populations for their in situ conservation in Northwestern Argentina

In situ conservation of wild species is a method of conservation that allows keeping populations in their natural environments, and set the strategies for maintaining the natural populations. The Active Bank of Northwestern Argentina (BANOA) is in charge of the in situ conservation of wild populations of common bean (Phaseolus vulgaris L.) in Northwestern Argentina (NOA), and has an ex situ collection of 401 landraces and 221 wild accessions from the NOA. We evaluated the phenotypic diversity of 68 common bean wild populations from the NOA both in protected and unprotected areas, finding a moderate variation among them. Ten phenotypic reproductive characteristics related to pod and seed displayed significant differences in the analysis of variance; these traits together with the seed weight were the basis for the multivariate analysis. The cluster analysis ordered the populations in 12 groups but trends in geographical distribution or phenotypical variation were not recognized. For the conservation in situ of the wild bean populations, their diversity should be considered. Two types of populations can be highlighted: (i) candidates for in situ conservation in order to preserve the novel variation generated by convergence with cultivated sympatric germplasm (populations 433, 437, 471, 509, 513 and 517) and (ii) those whose phenotype represents clearly the wild status and should be preserved in situ as such in their current status (populations 480, 495, 496, 525 and 533).     

Heterosis and combining ability of seven maize germplasm populations

Understanding the combining ability and heterosis of available germplasm is a prerequisite for successful maize improvement and breeding. The objectives of this study were to analyze the combining ability and heterosis of seven representative maize germplasm populations, and further, to evaluate their potential utility in germplasm improvement. A total of 21 crosses were made among these seven populations in a complete diallel without reciprocals. The parental populations and 21 crosses were evaluated for days to silking (DS), ear height (EH), and grain yield (GY) in the Northeast and Yellow and Huai River maize growing areas in China in 2012. Csyn5, Csyn7, Cpop.11, and Cpop.12 had desirable general combining ability (GCA) effects for DS and EH in both the Northeast China mega-environment (NCM) and the Yellow and Huai River Regions of China mega-environment (YHCM). Cpop.11 possessed a favorable GCA effect for GY in the NCM, as did Csyn5, Cpop.17, and Cpop.18 in the YHCM. Csyn6 and Csyn7 exhibited tremendous yield-enhancing potential in both mega-environments. Additionally, six combinations including Csyn7 × Csyn6, Csyn5 × Csyn6, Cpop.11 × Cpop.18, Cpop.12 × Cpop.17, Csyn7 × Cpop.17, and Csyn5 × Csyn7 exhibited better specific combining ability effects for GY, yield performance, and mid-parent heterosis in the appropriate mega-environment. These results indicated that the seven populations would be very useful for the improvement of related agronomic traits, and the six candidate combinations possessed great potential for further improvement and utilization.     

Genetics and molecular mapping of the naked grains in hexaploid oat

Avena sativa L. subsp. nudisativa has the ability to produce naked grains. Genetic studies on the naked trait of oat began over a century ago, but the genetic and molecular factors associated with the expression of this trait have not been fully clarified. The objectives of this study were to evaluate the naked trait in two oat populations of recombinant inbred lines (RILs), to determine the number of genes, to estimate the heritability, and to map genomic regions associated with the naked trait in hexaploid oat. Parental lines and RILs of each population were screened for the naked trait from plants grown in the field over a 2 year period. Based on the phenotypic data, the oat RILs were classed as naked, partially naked, partially hulled and hulled. In both populations and years, a great number of RILs showed variable expressivity for the naked trait. The genetic analysis indicated the action of a major gene (N1) with the action of modifying genes controlling the formation of naked grains. The results of the estimate of heritability show that environmental conditions do not have a great influence in determining the naked trait. The quantitative trait loci analysis detected a genomic region with a large effect on the naked trait that explained more than 50% of the phenotypic variation. Further studies are needed to validate the use of these molecular markers to assist breeding programs to select high quality and stable naked oat cultivars.     

Transformation of <Emphasis Type="Italic">Campanula</Emphasis> by wild type <Emphasis Type="Italic">Agrobacterium rhizogenes</Emphasis>

Compact growth is an important quality criterion in horticulture. Many Campanula species and cultivars exhibit elongated growth which is suppressed by chemical retardation and cultural practice during production to accommodate to the consumer’s desire. The production of compact plants via transformation with wild type Agrobacterium rhizogenes is an approach with great potential to produce plants that are non-GMO. Efficient transformation and regeneration procedures vary widely among both plant genera and species. Here we present a transformation protocol for Campanula. Hairy roots were produced on 26–90% of the petioles that were used for transformation of C. portenschlagiana (Cp), a C. takesimana × C. punctata hybrid (Chybr) and C. glomerata (Cg). Isolated hairy roots grew autonomously and vigorously without added hormones. The Cg hairy roots produced chlorophyll and generated plantlets in response to treatments with cytokinin (42 µM 2iP) and auxin (0.67 µM NAA). In contrast, regeneration attempts of transformed Cp and Chybr roots lead neither to the production of chlorophyll nor to the regeneration of shoots. Agropine A. rhizogenes strains integrate split T-DNA in T_L- and T_R-DNA fragments into the plant genome. In this study, regenerated plants of Cg did not contain T_R-DNA, indicating that a selective pressure against this T-DNA fragment may exist in Campanula.