The selection of superior walnut (<Emphasis Type="Italic">Juglans regia</Emphasis> L.) genotypes as revealed by morphological characterization

Walnut (Juglans regia L.), is one of the main economic fruit species in the world and is rich in nutritious substances. In order to obtain superior walnut genotypes, a set of seedling walnut genotypes were evaluated and their results were recorded. The range of nut length was between 25.57 and 47.77 mm. Also, the ranges of nut weight, kernel weight and kernel percentage varied from 5.35 to 21.31 g, 2.49 to 11.15 g and 37.27 to 66.29%, respectively. Simple correlation analysis showed significant positive and negative correlations between some of the measured traits. Principal component analysis reduced the evaluated traits into the seven main components, which explained 71.71% of the total variance. Bi-plot analysis performed using the first two components, distributed the genotypes into four sides of the plot. Based on the present results, a high variation was observed among the trees, indicating a high potential of the studied germplasm to select the superior genotypes. According to the present research, 57 genotypes with having the highest nut weight and kernel weight, as well as bright color of the kernel, were recognized as superior genotypes and can be used to establish the new orchards or improve cultivars.     

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.     

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.     

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 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.     

<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.     

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.     

Identification of quantitative trait loci involved in resistance to <Emphasis Type="Italic">Pseudomonas syringae</Emphasis> pv. <Emphasis Type="Italic">syringae</Emphasis> in pea (<Emphasis Type="Italic">Pisum sativum</Emphasis> L.)

Pseudomonas syringae is the main pathogen responsible for bacterial blight disease in pea and can cause yield losses of 70%. P. syringae pv. pisi is prevalent in most countries but the importance of P. syringae pv. syringae (Psy) is increasing. Several sources of resistance to Psy have been identified but genetics of the resistance is unknown. In this study the inheritance of resistance to Psy was studied in the pea recombinant inbred line population P665 × ‘Messire’. Results suggest a polygenic control of the resistance and two quantitative trait loci (QTL) associated with resistance, Psy1 and Psy2, were identified. The QTL explained individually 22.2 and 8.6% of the phenotypic variation, respectively. In addition 21 SSR markers were included in the P665 × ‘Messire’ map, of which six had not been mapped on the pea genome in previous studies.     

<Emphasis Type="Italic">Agrobacterium</Emphasis> mediated transformation of indica rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.) for insect resistance

The rice leaffolder (RLF), Cnaphalocrocis medinalis is an important pest of rice that causes severe damage in many areas of the world. The plants were transformed with fully modified (plant codon optimized) synthetic Cry1C coding sequences as well as with the hpt and gus genes, coding for hygromycin phosphotransferase and β-glucuronidase, respectively. Cry1C sequences placed under the control of doubled 35S promoter plus the AMV leader sequence, and hpt and gus genes driven by cauliflower mosaic virus 35S promoter, were used in this study. Embryogenic calli after cocultivation with Agrobacterium were selected on the medium containing hygromycin B. A total of 67 hygromycin-resistant plants were regenerated. PCR and Southern blot analyses of primary transformants revealed the stable integration of Cry1C coding sequences into the rice genome with predominant single copy integration. R₁ progeny plants disclosed a monogenic pattern (3:1) of transgene segregation as confirmed by molecular analyses. These transgenic lines were highly resistant to rice leaffolder (RLF), Cnaphalocrocis medinalis as revealed by insect bioassay.     

<Emphasis Type="Italic">Agrobacterium</Emphasis>-mediated genetic transformation of pigeon pea [<Emphasis Type="Italic">Cajanus cajan</Emphasis> (L.) Millsp.] for resistance to legume pod borer <Emphasis Type="Italic">Helicoverpa armigera</Emphasis>

Agrobacterium-mediated genetic transformation was performed using embryonic axes explants of pigeon pea. Both legume pod borer resistant gene (cry1Ac) and plant selectable marker neomycine phosphor transferase (nptII) genes under the constitutive expression of the cauliflower mosaic virus 35S promoter (CaMV35S) assembled in pPZP211 binary vector were used for the experiments. An optimum average of 44.61% successfully hardened dot blot Southern hybridization positive plants were obtained on co-cultivation media supplemented with 200 μM acetosyringone without L-cysteine. The increased transformation efficiency from a baseline of 11.53% without acetosyringone to 44.61% with acetosyringone was further declined with the addition of different concentrations of L-cysteine to co-cultivation media. Transgenic shoots were selected on 50 and 75 mg L⁻¹ kanamycin. Rooting efficiency was 100% on half-strength Murashige and Skoog medium with 20 g L⁻¹ sucrose and 0.5 mg L⁻¹ indole butyric acid in the absence of kanamycin. Furthermore, 100% seed setting was found among all the transgenic events. The plants obtained were subjected to multi- and nochoice tests to determine the behavioral responses and mortality through Helicoverpa armigera bioassays on the leaf and relate their relationship with the expression of cry1Ac protein which was found to be less in leaf as compared to the floral buds, anther, pod, and seed.     

Epistasis and heritability of resistance to <Emphasis Type="Italic">Phytophthora nicotianae</Emphasis> in pepper (<Emphasis Type="Italic">Capsicum annuum</Emphasis> L.)

Gene effects of resistance to two isolates of Phytophthora nicotianae in two crosses of pepper were investigated using separate generation means analysis. Additive-dominance models were inadequate in all cases. Digenic parameter models were adequate in three cases and the probability of goodness of fit of models was negatively correlated with the aggressiveness of the pathogen. None of these models explained variation among generation means in the combined cross Beldi × CM334 with P. nicotianae isolate Pn_2. Additive × additive, dominance × dominance and dominance × additive effects were significant in most cases. Additive and dominance effects (of negative sign) contribute more to resistance than to susceptibility. Additive variance was greater than environmental and dominance variance and ranged from 0.038 to 0.224. Narrow-sense heritabilities were dependent upon the cross and inoculate and ranged from 86 to 92%. The results of this study indicate that selection with more aggressive isolates of the pathogen will be useful for enhancing resistance in pepper.     

Interspecific hybridization of fig (<Emphasis Type="Italic">Ficus carica</Emphasis> L.) and <Emphasis Type="Italic">Ficus erecta</Emphasis> Thunb., a source of Ceratocystis canker resistance

Ceratocystis canker, which is caused by the fungus Ceratocystis fimbriata Ellis et Halsted, is one of the most severe diseases of the common fig (Ficus carica L.). In contrast, the wild fig species F. erecta Thunb. is resistant to this fungus. We performed interspecific hybridization between the common fig (seed parent) and F. erecta (pollen parent) through artificial pollination. Even though hybrid seeds showed high germination rates, the seedling survival rates were low. All of the seedlings contained the expected simple sequence repeat (SSR) alleles from both common fig and F. erecta at each of the three loci tested, thus confirming the parent–offspring relationships of the interspecific hybrids. The leaf morphological characters of hybrid seedlings were intermediate between those of the parents. Cuttings of cultivars of common fig, F. erecta, and hybrid seedlings were inoculated with C. fimbriata by direct wounding of the shoot. All of the common fig cultivars tested withered and died within 10 weeks. Leaves and shoots of the hybrids and F. erecta were healthy 100 days after inoculation. Our results suggest that interspecific hybridization between the common fig and the wild species F. erecta is a breakthrough in the breeding of a new fig rootstock source with resistance to Ceratocystis canker.     

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.     

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.     

Genome-wide SNP-based association mapping of resistance to <Emphasis Type="Italic">Phytophthora sojae</Emphasis> in soybean (<Emphasis Type="Italic">Glycine max</Emphasis> (L.) Merr.)

Phytophthora root rot (PRR) is among the most important soybean (Glycine max (L.) Merr.) diseases worldwide, and the host displays complex genetic resistance. A genome-wide association study was performed on 337 accessions from the Yangtze-Huai soybean breeding germplasm to identify resistance regions associated with PRR resistance using 60,862 high-quality single nucleotide polymorphisms markers. Twenty-six significant SNP-trait associations were detected on chromosomes 01 using a mixed linear model with the Q matrix and K matrix as covariates. In addition, twenty-six SNPs belonged to three adjacent haplotype blocks according to a linkage disequilibrium blocks analysis, and no previous studies have reported resistance loci in this 441 kb region. The real-time RT-PCR analysis of the possible candidate genes showed that two genes (Glyma01g32800 and Glyma01g32855) are likely involved in PRR resistance. Markers associated with resistance can contribute to marker-assisted selection in breeding programs. Analyses of candidate genes can lay a foundation for exploring the mechanism of P. sojae resistance.     

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.     

QTL analysis for thousand-grain weight under terminal drought stress in bread wheat (<Emphasis Type="Italic">Triticum</Emphasis><Emphasis Type="Italic">aestivum</Emphasis> L.)

Grain yield under post-anthesis drought stress is one of the most complex traits, which is inherited quantitatively. The present study was conducted to identify genes determining post-anthesis drought stress tolerance in bread wheat through Quantitative Trait Loci (QTLs) analysis. Two cultivated bread wheat accessions were selected as parental lines. Population phenotyping was carried out on 133 F_2:3 families. Two field experiments and two experiments in the greenhouse were conducted at IPK-Gatersleben, Germany with control and post-anthesis stress conditions in each experiment. Thousand-grain weight was recorded as the main wheat yield component, which is reduced by post-anthesis drought stress. Chemical desiccation was applied in three experiments as simulator of post-anthesis drought stress whereas water stress was applied in one greenhouse experiment. Analysis of variance showed significant differences among the F_2:3 families. The molecular genetic linkage map including 293 marker loci associated to 19 wheat chromosomes was applied for QTL analysis. The present study revealed four and six QTLs for thousand-grain weight under control and stress conditions, respectively. Only one QTL on chromosome 4BL was common for both conditions. Five QTLs on chromosomes 1AL, 4AL, 7AS, and 7DS were found to be specific to the stress condition. Both parents contributed alleles for drought tolerance. Taking the known reciprocal translocation of chromosomes 4AL/7BS into account, the importance of the short arms of homoeologous group 7 is confirmed for drought stress.     

Characterization of genes <Emphasis Type="Italic">Rpp2</Emphasis>, <Emphasis Type="Italic">Rpp4</Emphasis>, and <Emphasis Type="Italic">Rpp5</Emphasis> for resistance to soybean rust

Asian rust, caused by the fungus Phakopsora pachyrhizi, is the most severe disease currently threatening soybean crops in Brazil. The development of resistant cultivars is a top priority. Genetic characterization of resistance genes is important for estimating the improvement when these genes are introduced into soybean plants and for planning breeding strategies against this disease. Here, we infected an F₂ population of 140 plants derived from a cross between ‘An-76’, a line carrying two resistance genes (Rpp2 and Rpp4), and ‘Kinoshita’, a cultivar carrying Rpp5, with a Brazilian rust population. We scored six characters of rust resistance (lesion color [LC], frequency of lesions having uredinia [%LU], number of uredinia per lesion [NoU], frequency of open uredinia [%OU], sporulation level [SL], and incubation period [IP]) to identify the genetic contributions of the three genes to these characters. Furthermore, we selected genotypes carrying these three loci in homozygosis by marker-assisted selection and evaluated their genetic effect in comparison with their ancestors, An-76, PI230970, PI459025, Kinoshita and BRS184. All three genes contributed to the phenotypes of these characters in F₂ population and when pyramided, they significantly contributed to increase the resistance in comparison to their ancestors. Rpp2, previously reported as being defeated by the same rust population, showed a large contribution to resistance, and its resistance allele seemed to be recessive. Rpp5 had the largest contribution among the three genes, especially to SL and NoU. Only Rpp5 showed a significant contribution to LC. No QTLs for IP were detected in the regions of the three genes. We consider that these genes could contribute differently to resistance to soybean rust, and that genetic background plays an important role in Rpp2 activity. All three loci together worked additively to increase resistance when they were pyramided in a single genotype indicating that the pyramiding strategy is one good breeding strategy to increase soybean rust resistance.     

An approach to DNA polymorphism screening in <Emphasis Type="Italic">SBEIIa</Emphasis> homeologous genes of polyploid wheat (<Emphasis Type="Italic">Triticum</Emphasis> L.)

The non-transgenic manipulation of starch properties in common wheat (Triticum aestivum L.) generally implies combining mutant alleles of the particular gene copies in all three subgenomes (A, B and D). The redundancy of the hexaploid wheat chromosome set substantially complicates the identification of recessive mutations and breeding. Nevertheless, naturally occurring or induced genetic polymorphism has already been successfully exploited for the production of waxy (GBSSI-deficient) and elevated amylose (SSIIa-deficient) wheats. However, in order to achieve the amylose content above 50% of wheat endosperm starch, it may be necessary to inactivate the starch branching enzyme (SBEIIa) isoforms, as the RNAi repression results and gene expression data strongly suggest. The identification of null SBEIIa alleles and their combination in a single genotype is therefore a promising approach to the production of non-transgenic high-amylose wheat; however, wheat SBEIIa polymorphism has not been characterized as of yet. In order to develop an approach to SBEIIa mutation screening, we sequenced the SBEIIa central region (exons 9–12) from the three subgenomes of common wheat cv. Chinese Spring and the A genome of diploid einkorn T. monococcum. The genome-specific primers were developed that amplify the exons downstream from intron 11 selectively from each homeologous gene. Using a single-stranded DNA conformation polymorphism (SSCP) approach, we screened 60 wheat cultivars, landraces, and rare species for naturally occurring SNPs in exons 12, 13 and 14 of the three SBEIIa homeologs. In total, 13 SNPs were discovered in the A and B wheat genomes. Two of these SNPs affect the amino acid sequences of SBEIIa isoforms and may change the enzyme functional properties. The presence of restriction site polymorphism at SNP positions enables their easy genotyping with CAPS assays. Our results indicate that the mining for naturally occurring sequence polymorphism in starch biosynthesis genes of wheat can be successfully performed at the DNA level, providing the starting point for a search for SBEIIa mutants at a larger scale.