Phenotypic screening of seed retention and histological analysis of the abscission zone in Festuca pratensis and Lolium perenne.

BACKGROUND: Seed retention is the basic prerequisite for seed harvest. However, only little breeding progress has been achieved for this trait in the major forage grasses. The aim of this study was to evaluate the potential of plant genetic resources of the important fodder grasses Festuca pratensis Huds. and Lolium perenne L. as source for seed retention in the breeding process. Furthermore, the morphology of the abscission zone, where shattering occurs, was studied on the cell tissue level in different developmental stages of contrasting accessions. RESULTS: 150 and 286 accessions of Festuca pratensis and Lolium perenne were screened for seed retention, respectively. Contrasting accessions were selected to be tested in a second year. We found a great variation in seed retention in Festuca pratensis and Lolium perenne, ranging from 13 to 71% (average: 35%) and 12 to 94% (average: 49%), respectively, in the first year. Seed retention was generally lower in the second year. Cultivars were within the accessions with highest seed retention in Festuca pratensis, but had lower seed retention than ecotypes in Lolium perenne. Field-shattered seeds had a lower thousand grain weight than retained seeds. Cell layers of the abscission zone appeared already in early seed stages and were nested within each other in accessions with high seed retention, while there were two to three superimposed layers in accessions with low seed retention. CONCLUSIONS: Plant genetic resources of Lolium perenne might be a valuable source for breeding varieties with high seed retention. However, simultaneous selection for high seed weight is necessary for developing successful commercial cultivars.

Phenotypic variation of root-system architecture under high P and low P conditions in potato (Solanum tuberosum L.).

BACKGROUND: Phosphorus (P) is an essential macronutrient required for plant metabolism and growth. Its acquisition by plants depends on the availability of dissolved P in the rhizosphere and on the characteristics of P uptake mechanisms such as root-system architecture (RSA). Compared to other crops, potato (Solanum tuberosum L.) has a relatively poor P acquisition efficiency. This is mainly due to its shallow and sparsely branched root system, resulting in a rather limited exploitable soil volume. Information about potato genotypes with RSA traits suitable to improve adaptation to nutrient scarcity is quite rare. Aim of this study is to assess phenotypic variation of RSA in a potato diversity set and its reactions to P deficiency. RESULTS: Only one out of 22 RSA-traits showed a significant increase under low-P conditions. This indicates an overall negative effect of P scarcity on potato root growth. Differences among genotypes, however, were statistically significant for 21 traits, revealing a high variability in potato RSA. Using a principal component analysis (PCA), we were able to classify genotypes into three groups with regard to their root-system size. Genotypes with both small and large root systems reacted to low-P conditions by in- or decreasing their relative root-system size to medium, whereas genotypes with an intermediate root system size showed little to no changes. CONCLUSIONS: We observed a huge variation in both the potato root system itself and its adaptation to P deficiency. This may enable the selection of potato genotypes with an improved root-zone exploitation. Eventually, these could be utilized to develop new cultivars adapted to low-P environments with better resource-use efficiencies.

Genetic dissection of early-season cold tolerance in sorghum: genome-wide association studies for seedling emergence and survival under field and controlled environment conditions.

KEY MESSAGE: A QTL on sorghum chromosome SBI-06 putatively improves field emergence under low-temperature conditions. Low temperatures decisively limit seedling emergence and vigor during early growth of sorghum and, thus, strongly impair geographical expansion. To broaden sorghum cultivation to temperate regions, the establishment of cold-tolerant genotypes is a prioritized breeding goal. The present study aims at the quantification of seedling emergence and survival under chilling temperatures and the detection of marker-trait associations controlling temperature-related seedling establishment. A diversity set consisting of 194 biomass sorghum lines was subjected to extensive phenotyping comprising field trials and controlled environment experiments. The final emergence percentage (FEP) under field conditions was significantly reduced under cold stress. Broad-sense heritability was h 2 = 0.87 for FEP in the field and h 2 = 0.93 for seedling survival rate (SR) under controlled conditions. Correlations between FEP in the field and under controlled conditions were low; higher correlations were observed between field FEP and SR in controlled environments. Genome-wide association studies (GWAS) were conducted using 44,515 single nucleotide polymorphisms (SNPs) and revealed eight regions with suggestive marker-trait associations for FEP and SR on chromosomes SBI-01, -02, -03, -06, -09, and -10 (p < 5.7 × 10-5) and a significant association on SBI-06 for field FEP (p < 2.9 × 10-6). Although not significant under controlled conditions, SR of genotypes carrying the minor allele on the field FEP quantitative trait loci (QTL) on SBI-06 was on average 13.1% higher, while FEP under controlled conditions was on average 9.7% higher with a linearly decreasing effect with increasing temperatures (R 2 = 0.82). Promising candidate genes putatively conferring seedling cold tolerance were identified.

Sorghum root-system classification in contrasting P environments reveals three main rooting types and root-architecture-related marker-trait associations.

Background and Aims: Roots facilitate acquisition of macro- and micronutrients, which are crucial for plant productivity and anchorage in the soil. Phosphorus (P) is rapidly immobilized in the soil and hardly available for plants. Adaptation to P scarcity relies on changes in root morphology towards rooting systems well suited for topsoil foraging. Root-system architecture (RSA) defines the spatial organization of the network comprising primary, lateral and stem-derived roots and is important for adaptation to stress conditions. RSA phenotyping is a challenging task and essential for understanding root development. Methods: In this study, 19 traits describing RSA were analysed in a diversity panel comprising 194 sorghum genotypes, fingerprinted with a 90-k single-nucleotide polymorphism (SNP) array and grown under low and high P availability. Key Results: Multivariate analysis was conducted and revealed three different RSA types: (1) a small root system; (2) a compact and bushy rooting type; and (3) an exploratory root system, which might benefit plant growth and development if water, nitrogen (N) or P availability is limited. While several genotypes displayed similar rooting types in different environments, others responded to P scarcity positively by developing more exploratory root systems, or negatively with root growth suppression. Genome-wide association studies revealed significant quantitative trait loci (P < 2.9 × 10-6) on chromosomes SBI-02, SBI-03, SBI-05 and SBI-09. Co-localization of significant and suggestive (P < 5.7 × 10-5) associations for several traits indicated hotspots controlling root-system development on chromosomes SBI-02 and SBI-03. Conclusions: Sorghum genotypes with a compact, bushy and shallow root system provide potential adaptation to P scarcity in the field by allowing thorough topsoil foraging, while genotypes with an exploratory root system may be advantageous if N or water is the limiting factor, although such genotypes showed highest P uptake levels under the artificial conditions of the present study.

Phosphate uptake kinetics and tissue-specific transporter expression profiles in poplar (Populus × canescens) at different phosphorus availabilities.

BACKGROUND: Phosphorus (P) is a major plant nutrient. It is transported into and allocated inside plants by four families of phosphate transporters (PHT1 to PHT4) with high or low affinity to phosphate. Here, we studied whole-plant P uptake kinetics and expression profiles of members of the PHT families under high, intermediate and low P availability in the woody crop poplar (Populus × canescens) in relation to plant performance. RESULTS: Poplars exhibited strong growth reduction and increased P use efficiency in response to lower P availabilities. The relative P uptake rate increased with intermediate and decreased with low P availability. This decrease was not energy-limited because glucose addition could not rescue the uptake. The maximum P uptake rate was more than 13-times higher in P-starved than in well-supplied poplars. The Km for whole-root uptake ranged between 26 µM and 20 µM in poplars with intermediate and low P availability, respectively. In well-supplied plants, only low uptake rate was found. The minimum concentration for net P uptake from the nutrient solution was 1.1 µM. All PHT1 members studied showed significant up-regulation upon P starvation and were higher expressed in roots than leaves, with the exception of PtPHT1;3. PtPHT1;1 and PtPHT1;2 showed root- and P starvation-specific expression. Various members of the PHT2, PHT3 and PHT4 families showed higher expression in leaves than in roots, but were unresponsive to P deprivation. Other members (PtPHT3;1, PtPHT3;2, PtPHT3;6, PtPHT4;6 to PtPHT4;8) exhibited higher expression in roots than in leaves and were in most cases up-regulated in response to P deficiency. CONCLUSIONS: Expression profiles of distinct members of the PHT families, especially those of PHT1 were linked with changes in P uptake and allocation at whole-plant level. The regulation was tissue-specific with lower P responsiveness in leaves than in roots. Uptake efficiency for P increased with decreasing P availability, but could not overcome a threshold of about 1 µM P in the nutrient solution. Because the P concentrations in soil solutions are generally in the lower micro-molar range, even below the apparent Km-values, our findings suggest that bare-rooted poplars are prone to suffer from P limitations in most environments.

Phylogeny, tissue-specific expression, and activities of root-secreted purple acid phosphatases for P uptake from ATP in P starved poplar.

Plants secrete purple acid phosphatases (PAPs) under phosphorus (P) shortage but the contribution of plant PAPs to P acquisition is not well understood. The goals of this study were to investigate comprehensively the transcription patterns of PAPs under P shortage in poplar (Populus × canescens), to identify secreted PAPs and to characterize their contribution to mobilize organic P. Phylogenetic analyses of the PAP family revealed 33 putative members. In this study, distinct, tissue-specific P responsive expression patterns could be shown for 23 PAPs in roots and leaves. Root-associated PAP activities were localized on the root surface by in-vivo staining. The activities of root-surface PAPs increased significantly under low P availability, but were suppressed by a PAP inhibitor and corresponded to elevated P uptake from ATP as an organic P source. By proteomic analyses of the root apoplast, we identified three newly secreted proteins under P shortage: PtPAP1 (Potri.005G233400) and two proteins with unknown functions (Potri.013G100800 and Potri.001G209300). Our results, based on the combination of transcriptome and proteome analyses with phosphatase activity assays, support that PtPAP1 plays a central role in enhanced P acquisition from organic sources, when the phosphate concentrations in soil are limited.

Root-System Architectures of Two Cuban Rice Cultivars with Salt Stress at Early Development Stages.

Soil salinity is a critical problem for rice production and is also often associated with phosphors (P) deficiency. Plant hormones, like brassinosteroids, were shown to play a role in plant responses to different stresses and are also expected to mitigate salt stress. The aim of this study was to compare shoot growth and root architecture traits of two rice cultivars (INCA LP-5 and Perla de Cuba) during early plant development in response to salt, P limitation and a brassinosteroid. Seeds were placed in (I) paper rolls for 7 days and (II) mini-rhizotrons for 21 days without or with salt (50 mM NaCl), without or with 24-epibrassinolide (10-6 M) pre-treatment, and with two levels of P (10 or 1 ppm). The root system of LP-5 was larger in size and extent, while the roots of Perla were growing denser. Salt affected mainly the size- and extent-related root characteristics and explained about 70% of the variance. The effect of P was more pronounced without salt treatment. In Perla, P supply reduced the salt effect on root growth. The brassinosteroid had hardly any effect on the development of the plants in both experiments. Due to the high dependence on experimental factors, root length and related traits can be recommended for selecting young rice cultivars regarding salt stress and P deprivation.

Genotype-Specific Differences in Phosphorus Efficiency of Potato (Solanum tuberosum L.).

Potato is considered to have a low phosphorus (P) efficiency compared to other crops. Therefore, P fertilization requirements are high. New cultivars with improved P efficiency may contribute to save limited mineral P sources and to reduce eutrophication of surface water bodies. The present study aims to characterize the P efficiency of different potato genotypes and to identify mechanisms that improve P efficiency in cultivated potato. A diversity set of 32 potato accessions was used to assess their P efficiency. From this set, five cultivars were selected and two pot experiments with different P-fertilization strategies including a non-fertilized control were conducted to estimate effects of P deficiency on general agronomic and P related traits, root development, phosphatase activity and micro RNA 399 (miR399) expression. Significant differences between the 32 genotypes were found for P utilization efficiency (PUtE). P acquisition efficiency (PAE) as P content in low P in relation to P content in high P was positively correlated to relative biomass production while PUtE was not. Selected genotypes displayed a strong relation between total root length and P content. Root phosphatase activity and miR399 expression increased under P deficiency. However, tuber yields of four cultivars, grown on a soil with suboptimal content of plant available P, were not significantly affected in comparison to yields of well-fertilized plots. We conclude from the present study that PUtE and PAE are important traits when selecting for plants requiring less fertilizer inputs but PAE might be more important for cropping on deficient soils. A large root system might be the most important trait for P acquisition on such soils and therefore in breeding for P efficient crops. Lowering P fertilizer inputs might not necessarily reduce tuber yields.

Dissecting nutrient-related co-expression networks in phosphate starved poplars.

Phosphorus (P) is an essential plant nutrient, but its availability is often limited in soil. Here, we studied changes in the transcriptome and in nutrient element concentrations in leaves and roots of poplars (Populus × canescens) in response to P deficiency. P starvation resulted in decreased concentrations of S and major cations (K, Mg, Ca), in increased concentrations of N, Zn and Al, while C, Fe and Mn were only little affected. In roots and leaves >4,000 and >9,000 genes were differently expressed upon P starvation. These genes clustered in eleven co-expression modules of which seven were correlated with distinct elements in the plant tissues. One module (4.7% of all differentially expressed genes) was strongly correlated with changes in the P concentration in the plant. In this module the GO term "response to P starvation" was enriched with phosphoenolpyruvate carboxylase kinases, phosphatases and pyrophosphatases as well as regulatory domains such as SPX, but no phosphate transporters. The P-related module was also enriched in genes of the functional category "galactolipid synthesis". Galactolipids substitute phospholipids in membranes under P limitation. Two modules, one correlated with C and N and the other with biomass, S and Mg, were connected with the P-related module by co-expression. In these modules GO terms indicating "DNA modification" and "cell division" as well as "defense" and "RNA modification" and "signaling" were enriched; they contained phosphate transporters. Bark storage proteins were among the most strongly upregulated genes in the growth-related module suggesting that N, which could not be used for growth, accumulated in typical storage compounds. In conclusion, weighted gene coexpression network analysis revealed a hierarchical structure of gene clusters, which separated phosphate starvation responses correlated with P tissue concentrations from other gene modules, which most likely represented transcriptional adjustments related to down-stream nutritional changes and stress.