Despite the fact that Fv/Fm (maximum quantum efficiency of photosystem II) is the most widely used parameter for a rapid non-destructive measure of stress detection in plants, there are barely any studies on the genetic understanding of this trait under heat stress. Our aim was to identify quantitative trait locus (QTL) and the potential candidate genes linked to Fv/Fm for improved photosynthesis under heat stress in wheat (Triticum aestivum L.). Three bi-parental F2 mapping populations were generated by crossing three heat tolerant male parents (origin: Afghanistan and Pakistan) selected for high Fv/Fm with a common heat susceptible female parent (origin: Germany) selected for lowest Fv/Fm out of a pool of 1274 wheat cultivars of diverse geographic origin. Parents together with 140 F2 individuals in each population were phenotyped by Fv/Fm under heat stress (40°C for 3 days) around anthesis. The Fv/Fm decreased by 6.3% in the susceptible parent, 1–2.5% in the tolerant parents and intermediately 4–6% in the mapping populations indicating a clear segregation for the trait. The three populations were genotyped with 34,955 DArTseq and 27 simple sequence repeat markers, out of which ca. 1800 polymorphic markers mapped to 27 linkage groups covering all the 21 chromosomes with a total genome length of about 5000 cM. Inclusive composite interval mapping resulted in the identification of one significant and heat-stress driven QTL in each population on day 3 of the heat treatment, two of which were located on chromosome 3B and one on chromosome 1D. These QTLs explained about 13–35% of the phenotypic variation for Fv/Fm with an additive effect of 0.002–0.003 with the positive allele for Fv/Fm originating from the heat tolerant parents. Approximate physical localization of these three QTLs revealed the presence of 12 potential candidate genes having a direct role in photosynthesis and/or heat tolerance. Besides providing an insight into the genetic control of Fv/Fm in the present study, the identified QTLs would be useful in breeding for heat tolerance in wheat.

Several researchers have hypothesized that shade avoidance behaviour is favoured by natural selection because it increases the fitness of individuals. Shade avoidance can be disadvantageous for crops, however, because it reduces allocation of resources to reproductive yield, increases the risk of lodging and reduces weed suppression. One approach to develop varieties with reduced shade avoidance and enhanced agronomic performance is by inducing mutations followed by phenotypic screening. We treated spring wheat seeds with ethyl methanesulfonate and screened the seedlings repeatedly under green filters for plants showing reduced elongation of the first leaf sheath and second leaf lamina. The shade avoidance responses of five promising mutant lines were further compared to non-mutated plants in a climate chamber experiment with added far-red light. Two of the selected lines displayed significantly reduced elongation under all light treatments while two lines showed reduced elongation only in added far-red light. The most promising mutant line did not differ in height from the non-mutated cultivar in neutral light, but elongated 20.6% less in strong far-red light. This traditional forward approach of screening mutagenized spring wheat produced plants with reduced shade avoidance responses. These mutants may generate new molecular handles to modify the reaction of plants to changes in light spectral distribution in traditional and novel cultivation systems.

The ability to evolve novel metabolites has been instrumental for the defence of plants against antagonists. A few species in the Barbarea genus are the only crucifers known to produce saponins, some of which make plants resistant to specialist herbivores, like Plutella xylostella, the diamondback moth. Genetic mapping in Barbarea vulgaris revealed that genes for saponin biosynthesis are not clustered but are located in different linkage groups. Using co-location with quantitative trait loci (QTLs) for resistance, transcriptome and genome sequences, we identified two 2,3-oxidosqualene cyclases that form the major triterpenoid backbones. LUP2 mainly produces lupeol, and is preferentially expressed in insect-susceptible B. vulgaris plants, whereas LUP5 produces β-amyrin and α-amyrin, and is preferentially expressed in resistant plants; β-amyrin is the backbone for the resistance-conferring saponins in Barbarea. Two loci for cytochromes P450, predicted to add functional groups to the saponin backbone, were identified: CYP72As co-localized with insect resistance, whereas CYP716As did not. When B. vulgaris sapogenin biosynthesis genes were transiently expressed by CPMV-HT technology in Nicotiana benthamiana, high levels of hydroxylated and carboxylated triterpenoid structures accumulated, including oleanolic acid, which is a precursor of the major resistance-conferring saponins. When the B. vulgaris gene for sapogenin 3-O-glucosylation was co-expressed, the insect deterrent 3-O-oleanolic acid monoglucoside accumulated, as well as triterpene structures with up to six hexoses, demonstrating that N. benthamiana further decorates the monoglucosides. We argue that saponin biosynthesis in the Barbarea genus evolved by a neofunctionalized glucosyl transferase, whereas the difference between resistant and susceptible B. vulgaris chemotypes evolved by different expression of oxidosqualene cyclases (OSCs).

The chlorophyll fluorescence parameter Fv /Fm reflects the maximum quantum efficiency of photosystem II (PSII) photochemistry and has been widely used for early stress detection in plants. Previously, we have used a three-tiered approach of phenotyping by Fv /Fm to identify naturally existing genetic variation for tolerance to severe heat stress (3 days at 40°C in controlled conditions) in wheat (Triticum aestivum L.). Here we investigated the performance of the previously selected cultivars (high and low group based on Fv /Fm value) in terms of growth and photosynthetic traits under moderate heat stress (1 week at 36/30°C day/night temperature in greenhouse) closer to natural heat waves in North-Western Europe. Dry matter accumulation after 7 days of heat stress was positively correlated to Fv /Fm . The high Fv /Fm group maintained significantly higher total chlorophyll and net photosynthetic rate (PN ) than the low group, accompanied by higher stomatal conductance (gs ), transpiration rate (E) and evaporative cooling of the leaf (ΔT). The difference in PN between the groups was not caused by differences in PSII capacity or gs as the variation in Fv /Fm and intracellular CO2 (Ci ) was non-significant under the given heat stress. This study validated that our three-tiered approach of phenotyping by Fv /Fm performed under increasing severity of heat was successful in identifying wheat cultivars differing in photosynthesis under moderate and agronomically more relevant heat stress. The identified cultivars may serve as a valuable resource for further studies to understand the physiological mechanisms underlying the genetic variability in heat sensitivity of photosynthesis.

A collection of 100 wheat varieties representing more than 100 years of wheat-breeding history in Scandinavia was established in order to identify marker-trait associations for plant height (PH), grain yield (GY), and biomass potential for bioethanol production. The field-grown material showed variations in PH from 54 to 122 cm and in GY from 2 to 6.61 t ha(-1). The release of monomeric sugars was determined by high-throughput enzymatic treatment of ligno-cellulosic material and varied between 0.169 and 0.312 g/g dm for glucose (GLU) and 0.146 and 0.283 g/g dm for xylose (XYL). As expected, PH and GY showed to be highly influenced by genetic factors with repeatability (R) equal to 0.75 and 0.53, respectively, while this was reduced for GLU and XYL (R = 0.09 for both). The study of trait correlations showed how old, low-yielding, tall varieties released higher amounts of monomeric sugars after straw enzymatic hydrolysis, showing reduced recalcitrance to bioconversion compared to modern varieties. Ninety-three lines from the collection were genotyped with the DArTseq(®) genotypic platform and 5525 markers were used for genome-wide association mapping. Six quantitative trait loci (QTLs) for GY, PH, and GLU released from straw were mapped. One QTL for PH was previously reported, while the remaining QTLs constituted new genomic regions linked to trait variation. This paper is one of the first studies in wheat to identify QTLs that are important for bioethanol production based on a genome-wide association approach.

The genotypic response of wheat cultivars as affected by two methods of heat stress treatment (treatment of intact plants in growth chambers versus treatment of detached leaves in test tubes) in a temperature controlled water bath were compared to investigate how such different methods of heat treatment affect chlorophyll fluorescence parameters. A set of 41 spring wheat cultivars differing in their maximum photochemical efficiency of photosystem (PS) II (Fv/Fm) under heat stress conditions was used. These cultivars were previously evaluated based on the heat treatment of intact plants. The responses of the same cultivars to heat stress were compared between the two methods of heat treatment. The results showed that in detached leaves, all of the fluorescence parameters remained almost unaffected in control (20°C at all durations tested), indicating that the detachment itself did not affect the fluorescence parameters. In contrast, heat induced reduction in the maximum photochemical efficiency of PSII of detached leaves occurred within 2h at 40°C and within 30min at 45°C, and the response was more pronounced than when intact plants were heat stressed for three days at 40°C. The proportion of total variation that can be ascribed to the genetic differences among cultivars for a trait was estimated as genetic determination. During heat treatment, the genetic determination of most of the fluorescence parameters was lower in detached leaves than in intact plants. In addition, the correlation of the cultivar response in intact plants versus detached leaves was low (r=0.13 (with expt.1) and 0.02 with expt.2). The most important difference between the two methods was the pronounced difference in time scale of reaction, which may indicate the involvement of different physiological mechanisms in response to high temperatures. Further, the results suggest that genetic factors associated with cultivar differences are different for the two methods of heat treatment.

Duplicate accessions in gene banks may be increasing while funding resources to maintain them are not always available. This research investigated the ability of agronomic and morphological descriptors for detecting duplicates among 138 repatriated putative Nordic barley germplasm and compared their use with results from previous research with microsatellite markers. These accessions were initially grouped into 36 potential duplicates according to passport data but further analysis with microsatellites reduce them to 22 genetically homogeneous groups. The analysis with 26 agronomic and morphological descriptors of putative Nordic spring barley accessions from nine gene banks was compared with a previous study with microsatellites. Each agronomic and morphological descriptor was weighed relative to its genetic determination with the aim of reducing the effect of environmental errors on genetic distances. Nonetheless, weighted and non-weighted genetic distances were significantly correlated (r = 0.956) but their respective correlations with genetic distances based on microsatellites were smaller (r = 0.526 and r = 0.498, respectively). Genetic distances based on agronomic and morphological descriptors were smaller than those based on microsatellites. The genetic distances based on agronomic and morphological descriptors showed a bimodal distribution, which was apparently due to the number of rows per spike. Our research demonstrated, nonetheless, the ability of genetic distances based on agronomic and morphological discriminators to detect genetic heterogeneity, although with reduced sensitivity compared with microsatellite markers. These results also suggest that grouping ensuing from either approach could reflect distinct patterns of diversity (due to different mutation rates and selection intensity) and are therefore complementary.

Landraces of seed and dessert type watermelons from three provinces of Mozambique with distinct agro-ecological conditions, including humid and semi-dry areas from the north to the south of the country, were analysed to assess their genetic differentiation. Ninety-six accessions (269 plants) were tested with 24 microsatellite markers resulting in 110 alleles. Analysis of molecular variance showed that 63% of the total variation in the plant material could be explained among the accessions, while 37% of the variation was within accessions. Molecular variance between material used for seed extraction and dessert consumption explained 34% of the total variation whereas villages explained 27% of the molecular variation among the sampled locations. Structure analysis revealed that the material could be differentiated into three genetic groups. The seed types clustered in one genetic group, irrespective of provincial origin. Dessert type accessions from the semi-arid south and the central part were assigned to a second genetic group, while accessions from the northern more humid region of the country were assigned to a third genetic group. The observed genetic diversity may reflect farmer selection under different agro-ecological conditions or an introduction of material from different sources into the growing areas. Keywords: Citrullus lanatus , DNA fingerprinting, landraces, Mozambique, microsatellites, simple sequence repeats (SSR). African Journal of Biotechnology Vol. 12(36), pp. 5513-5521

This paper undertakes an exploration of the potential application of the Weitzman prioritisation approach to the conservation of plant genetic resources (PGR). Using a case study approach and assuming limited conservation resources, this approach is applied to calculate the maximum diversity which can be conserved based on a clustering of cacao species (Theobroma cacao L). A conservation budget allocation model applied across a set of ten clusters and nine subclusters of cacao, together with the use of alternative diversity and risk measures, allowed for an evaluation of a range of potential conservation outcomes. Alternative risk measures generally resulted in the allocation of conservation funds to the same priority clusters of cacao (Criollo and Curaray). However, the use of the number of locally common alleles as an alternative to the original Weitzman diversity measure produced a markedly different result, prioritising the Puri's cluster in Western Amazonia. The Curaray cluster was highly prioritised under both diversity measures, indicating its distinctiveness relative to both the Criollo and Puri's clusters. We conclude that the Weitzman approach can indeed allow diversity comparisons to be made between the outcomes of different PGR conservation strategies, although subjective decisions regarding the choice of diversity and risk measures remain. (C) 2013 Elsevier B.V. All rights reserved.

With an objective to evaluate the Bhutan rice (Oryza sativa L.) landraces for genetic diversity in blast resistance, 402 accessions viz. 352 landraces, a differential set of 32 R gene lines and 18 modern cultivars were field-evaluated in three blast 'hotspot' sites followed by genetic analysis of the 352 landraces with 27 microsatellite markers. Across the sites, 19 landraces (5.4%) exhibited complete resistance with zero disease score and 203 (58%) and 163 (46%) landraces showed high partial resistance for early leaf and panicle blast, respectively, with disease score of four to six. Field evaluation for leaf and panicle blast at three experimental sites in Bhutan showed best cultivar discrimination in early leaf blast at the tillering stage (heritability, h(2) = 0.32) and in the panicle blast at maturity (h(2) = 0.44). Subdivision of the genetic variation into cultivar groups revealed the most variation for blast severity within the 352 landraces with h(2) of 0.31 and 0.46 for early leaf and panicle blast, respectively. Cluster analysis of the landraces revealed two distinct rice cultivar groups, which separated at dissimilarity of 0.84 according to origin of the cultivars from low, mid and high altitude zones in Bhutan. All microsatellites were polymorphic with two to 21 different alleles per marker and a high polymorphic information content value of 0.61. The identified blast resistant landraces were genetically diverse originating from different rice cultivation zones. Further investigation of the resistant and partial resistant material may reveal specific blast resistance genes, which could be useful to mitigate blast incidence in rice-producing countries.

In view of the global climate change, heat stress is an increasing constraint for the productivity of wheat (Triticum aestivum L.). Our aim was to identify contrasting cultivars in terms of heat tolerance by mass screening of 1274 wheat cultivars of diverse origin, based on a physiological trait, the maximum quantum efficiency of PSII (Fv/Fm). A chlorophyll fluorescence protocol was standardised and used for repeated screening with increased selection pressure with a view to identifying a set of cultivars extreme for the trait. An initial mass screening of 1274 wheat cultivars with a milder heat stress of 38°C in 300µmolm-2s-1 for 2h with preheating at 33-35°C for 19h in 7-14µmolm-2s-1 light showed a genetic determination of 8.5±2.7%. A heat treatment of 40°C in 300µmolm-2s-1 for 72h in the second screening with 138 selected cultivars resulted in larger differentiation of cultivars with an increased genetic component (15.4±3.6%), which was further increased to 27.9±6.8% in the third screening with 41 contrasting cultivars. This contrasting set of cultivars was then used to compare the ability of chlorophyll fluorescence parameters to detect genetic difference in heat tolerance. The identification of a set of wheat cultivars contrasting for their inherent photochemical efficiency may aid future studies to understand the genetic and physiological nature of heat stress tolerance in order to dissect quantitative traits into simpler genetic factors.

The degradability of cereal straw is of importance when it is used for animal feed, biological means of bioenergy production such as bioethanol production and when it is incorporated in soil. We examined wheat straw from 106 different winter wheat cultivars representing the northwest European contemporary gene pool. The cultivars were grown at two different locations to assess the potential for breeding for improved degradability. The straws exhibited much variation in degradability ranging from 258 g kg(-1) to 407 g kg(-1) of dry matter. The heritability for degradability was estimated to 29% indicating a reasonable potential for response to selection. Inclusion of height as a regression-term, indicated that only a minor part of genetic differences are directly related to plant height and that improvements in degradability may be achieved without unacceptable changes in straw length. Finally, a lack of correlation between degradability and grain yield indicated that straw degradability may be improved through breeding without serious negative effect on grain yield. (C) 2010 Elsevier Ltd. All rights reserved.

Saponins are bioactive compounds generally considered to be produced by plants to counteract pathogens and herbivores. Besides their role in plant defense, saponins are of growing interest for drug research as they are active constituents of several folk medicines and provide valuable pharmacological properties. Accordingly, much effort has been put into unraveling the modes of action of saponins, as well as in exploration of their potential for industrial processes and pharmacology. However, the exploitation of saponins for bioengineering crop plants with improved resistances against pests as well as circumvention of laborious and uneconomical extraction procedures for industrial production from plants is hampered by the lack of knowledge and availability of genes in saponin biosynthesis. Although the ability to produce saponins is rather widespread among plants, a complete synthetic pathway has not been elucidated in any single species. Current conceptions consider saponins to be derived from intermediates of the phytosterol pathway, and predominantly enzymes belonging to the multigene families of oxidosqualene cyclases (OSCs), cytochromes P450 (P450s) and family 1 UDP-glycosyltransferases (UGTs) are thought to be involved in their biosynthesis. Formation of unique structural features involves additional biosynthetical enzymes of diverse phylogenetic background. As an example of this, a serine carboxypeptidase-like acyltransferase (SCPL) was recently found to be involved in synthesis of triterpenoid saponins in oats. However, the total number of identified genes in saponin biosynthesis remains low as the complexity and diversity of these multigene families impede gene discovery based on sequence analysis and phylogeny. This review summarizes current knowledge of triterpenoid saponin biosynthesis in plants, molecular activities, evolutionary aspects and perspectives for further gene discovery.

Breeding better cultivars has become a highly efficient way to improve plant production for yield, quality and reduced input. Most breeding activities have been focussed into fewer and larger programs with less people engaged. Educational environments for the area have been correspondingly reduced in size, which makes it harder to provide inspiring environments to generate high level education, for the fewer people needed in industry. Still plant breeders, scientists as well as society have ample interest in widespread public understanding of the use of new as well as old technologies for improvement of our cultivated plants. This is not least to avoid future communication problems with the general public like experienced with genetically modified plants during recent years.

Evolutionary theory can be applied to improve agricultural yields and/or sustainability, an approach we call Evolutionary Agroecology. The basic idea is that plant breeding is unlikely to improve attributes already favored by millions of years of natural selection, whereas there may be unutilized potential in selecting for attributes that increase total crop yield but reduce plants' individual fitness. In other words, plant breeding should be based on group selection. We explore this approach in relation to crop-weed competition, and argue that it should be possible to develop high density cereals that can utilize their initial size advantage over weeds to suppress them much better than under current practices, thus reducing or eliminating the need for chemical or mechanical weed control. We emphasize the role of density in applying group selection to crops: it is competition among individuals that generates the 'Tragedy of the Commons', providing opportunities to improve plant production by selecting for attributes that natural selection would not favor. When there is competition for light, natural selection of individuals favors a defensive strategy of 'shade avoidance', but a collective, offensive 'shading' strategy could increase weed suppression and yield in the high density, high uniformity cropping systems we envision.

Combined genomics and metabolomics approaches were used to unravel molecular mechanisms behind interactions between winter cress (Barbarea vulgaris) and flea beetle (Phyllotreta nemorum). B. vulgaris comprises two morphologically, biochemically and cytologically deviating types, which differ in flea beetle resistance, saponin and glucosinolate profiles, as well as leaf pubescence. An F2 population generated from a cross between the two B. vulgaris types was used to construct a B. vulgaris genetic map based on 100 AFLP and 31 microsatellite markers. The map was divided into eight linkage groups. QTL (quantitative trait loci) analysis revealed a total of 15 QTL affecting eight traits, including nine QTL for four saponins, two QTL for two glucosinolates, two QTL for hairiness, and two QTL for flea beetle resistance. The two QTL for resistance towards flea beetles in B. vulgaris co-localized with QTL for the four saponins associated with resistance. Furthermore, global QTL analysis of B. vulgaris metabolites identified QTL for a number of flavonoid glycosides and additional saponins from both resistant and susceptible types. The transcriptome of the resistant B. vulgaris type was sequenced by pyrosequencing, and sequences containing microsatellites were identified. Microsatellite types in B. vulgaris were similar to Arabidopsis thaliana but different from Oryza sativa. Comparative analysis between B. vulgaris and A. thaliana revealed a remarkable degree of synteny between a large part of linkage groups 1 and 4 of B. vulgaris harboring the two QTL for flea beetle resistance and Arabidopsis chromosomes 3 and 1. Gene candidates that may underlie QTL for resistance and saponin biosynthesis are discussed.

Winter cress (Barbarea vulgaris) is resistant to a range of insect species. Some B. vulgaris genotypes are resistant, whereas others are susceptible, to herbivory by flea beetle larvae (Phyllotreta nemorum). Metabolites involved in resistance to herbivory by flea beetles were identified using an ecometabolomic approach. An F2 population representing the whole range from full susceptibility to full resistance to flea beetle larvae was generated by a cross between a susceptible and a resistant B. vulgaris plant. This F2 offspring was evaluated with a bioassay measuring the ability of susceptible flea beetle larvae to survive on each plant. Metabolites that correlated negatively with larvae survival were identified through correlation, cluster, and principal component analyses. Two main clusters of metabolites that correlate negatively with larvae survival were identified. Principal component analysis grouped resistant and susceptible plants as well as correlated metabolites. Known saponins, such as hederagenin cellobioside and oleanolic acid cellobioside, as well as two other saponins correlated significantly with plant resistance. This study shows the potential of metabolomics to identify bioactive compounds involved in plant defense.

The application of recombinant (His)(6)-tagged proteins in cell culture assays is associated with problems due to lipopolysaccharide (LPS) contamination. LPS stimulates cells of the immune system, thereby masking antigen-specific activation of T cells. Due to the affinity of LPS for histidine it is associated with difficulties to remove LPS from recombinant (His)(6)-tagged proteins. Here we describe that the Triton X-114 phase separation method can be used to remove LPS from (His)(6)-tagged proteins and that the recombinant proteins retain their biological activity.

ABSTRACTThe NOVA university network consists of a collaboration between agriculture‐related institutions in Sweden, Norway, Finland, Iceland, and Denmark with the goal of supporting veterinary, forestry, and agricultural education. The plant breeding part of these activities was already initiated with joint Ph.D. courses in 1975 and also includes Baltic institutions from Estonia, Latvia, and Lithuania since 1993. The joint courses and other network activities are a means of maintaining a true plant breeding identity in the geographical area while rapid changes take place within the agricultural industry, university curricula, and students' perception of the issue.