Search

Nitrogen (N) is an abundant element on earth. It is a crucial macro-element for plant growth and development. The lack of N causes reduced shoot growth and photosynthesis, early flowering, increased lateral/primary root ratio, and anthocyanin accumulation. Farmers apply N fertilizers to deal with N shortage, however, it causes adverse effects on the environment. Plants are adapted to live under low N supply and can still sustain growth and reproduction. N use efficiency (NUE), shoot dry matter per shoot N content, is used to measure the adaptation to the N depletion. This research investigates the candidate genes, which are responsible for sub-optimal N supply adaptation by using Arabidopsis thaliana, as a model organism. 354 diverse accessions of a natural population, called as HapMap population, are screened under sub-optimal (1 mM) and optimal (5 mM) N (in the form of ammonium/nitrate) conditions. NUE, related agronomic traits (NRATs), and phenotypic plasticity are used to map quantitative trait loci (QTLs). Genome-wide association study (GWAS) is used as a mapping technique and identifies 28 QTLs. 20 accessions from HapMap population are reassessed to confirm extreme accessions in the population. Besides, positive correlation is observed between NUE and relative growth rate and between shoot N concentration and water content. Br-0 and Col-0, two distinct accessions on NUE, are selected as parental accessions for further QTL investigation using bi-parental linkage mapping. QTL mapping reveals seven loci in Br-0 and Col-0 RIL population. One locus, associated with the phenotypic plasticity of water content in GWAS and the shoot fresh weight in the bi-parental linkage mapping, is found to be co-located in both mapping results. This research indicates that the natural genetic variation in Arabidopsis thaliana has a great potential for investigation of NUE improvement in plants. Both GWAS and bi-parental linkage mapping use the natural genetic variation to identify Q

Zn is an essential element for life and the second most abundant transition element in all organisms. Zn deficiency is one of most widespread micronutrient deficiencies in soils and plants need to cope with it. Plants are able to sense the shortage of Zn supply and adjust their Zn homeostasis accordingly. In this thesis, I explored the genetic architecture of the Zn deficiency response of Arabidopsis thaliana. I initiated functional analyses on some of the Zn transporter genes long known to be transcriptionally induced by Zn deficiency, but so far not investigated in much detail. Soon after the exposure to Zn deficiency the expression of these transporters is first induced in roots and later in shoots. Each Zn transporter gene is expressed in one, or more, specific root cell layer(s), which location may differ, depending on the root part. These transporters showed partial functional redundancy among them, which makes it difficult, if not impossible, to detect particular Zn deficiency phenotypes in single Zn transporter mutants. Considering that the effect of mutations on Zn transporter genes is difficult to detect, the Zn deficiency response can be studied from the perspective of their regulatory components. In the search of these regulators I found that the disruption of the N-ALPHA-TERMINAL ACETYLTRANSFERASE 25 (NAA25) gene, involved in N-terminal-acetylation of proteins, causes a strong Fe deficiency sensitivity and a slightly, but consistently, higher expression of several Zn deficiency responsive genes upon Zn sufficiency, when compared to wild-type plants. The causes for the unusual expression pattern of these genes still remain unknown. One hypothesis I postulate is that the lack of N-terminal acetylation of the Zn deficiency regulators bZIP19 and bZIP23, which are possible targets N-terminal acetylation, alters their regulatory activity. To identify more unconventional genetic players underlying the Zn deficiency response, that is not the known suspects invo

Contains fulltext : 163402.pdf (publisher's version ) (Open Access)

Contains fulltext : 163402.pdf (publisher's version ) (Open Access)

Contains fulltext : 163402.pdf (publisher's version ) (Open Access)

Contains fulltext : 126119.pdf (publisher's version ) (Open Access), Knowledge of the exact distribution of meiotic crossovers (COs) and gene conversions (GCs) is essential for understanding many aspects of population genetics and evolution, from haplotype structure and long-distance genetic linkage to the generation of new allelic variants of genes. To this end, we resequenced the four products of 13 meiotic tetrads along with 10 doubled haploids derived from Arabidopsis thaliana hybrids. GC detection through short reads has previously been confounded by genomic rearrangements. Rigid filtering for misaligned reads allowed GC identification at high accuracy and revealed an approximately 80-kb transposition, which undergoes copy-number changes mediated by meiotic recombination. Non-crossover associated GCs were extremely rare most likely due to their short average length of approximately 25-50 bp, which is significantly shorter than the length of CO-associated GCs. Overall, recombination preferentially targeted non-methylated nucleosome-free regions at gene promoters, which showed significant enrichment of two sequence motifs. DOI: http://dx.doi.org/10.7554/eLife.01426.001.

Contains fulltext : 126119.pdf (publisher's version ) (Open Access), Knowledge of the exact distribution of meiotic crossovers (COs) and gene conversions (GCs) is essential for understanding many aspects of population genetics and evolution, from haplotype structure and long-distance genetic linkage to the generation of new allelic variants of genes. To this end, we resequenced the four products of 13 meiotic tetrads along with 10 doubled haploids derived from Arabidopsis thaliana hybrids. GC detection through short reads has previously been confounded by genomic rearrangements. Rigid filtering for misaligned reads allowed GC identification at high accuracy and revealed an approximately 80-kb transposition, which undergoes copy-number changes mediated by meiotic recombination. Non-crossover associated GCs were extremely rare most likely due to their short average length of approximately 25-50 bp, which is significantly shorter than the length of CO-associated GCs. Overall, recombination preferentially targeted non-methylated nucleosome-free regions at gene promoters, which showed significant enrichment of two sequence motifs. DOI: http://dx.doi.org/10.7554/eLife.01426.001.

Contains fulltext : 126119.pdf (publisher's version ) (Open Access), Knowledge of the exact distribution of meiotic crossovers (COs) and gene conversions (GCs) is essential for understanding many aspects of population genetics and evolution, from haplotype structure and long-distance genetic linkage to the generation of new allelic variants of genes. To this end, we resequenced the four products of 13 meiotic tetrads along with 10 doubled haploids derived from Arabidopsis thaliana hybrids. GC detection through short reads has previously been confounded by genomic rearrangements. Rigid filtering for misaligned reads allowed GC identification at high accuracy and revealed an approximately 80-kb transposition, which undergoes copy-number changes mediated by meiotic recombination. Non-crossover associated GCs were extremely rare most likely due to their short average length of approximately 25-50 bp, which is significantly shorter than the length of CO-associated GCs. Overall, recombination preferentially targeted non-methylated nucleosome-free regions at gene promoters, which showed significant enrichment of two sequence motifs. DOI: http://dx.doi.org/10.7554/eLife.01426.001.

Arabidopsis thaliana is the plant species that in the past 25 years has developed into the major model species in plant biology research. This was due to its properties such as short generation time, its small genome and its easiness to be transformed. Wageningen University has played an important role in the development of this model, based on interdisciplinary collaborations using genetics as a major tool to investigate aspects of physiology, development, plant-microbe interactions and evolution.

Life-history traits controlling the duration and timing of developmental phases in the life cycle jointly determine fitness. Therefore, life-history traits studied in isolation provide an incomplete view on the relevance of life-cycle variation for adaptation. In this study, we examine genetic variation in traits covering the major life history events of the annual species Arabidopsis thaliana: seed dormancy, vegetative growth rate and flowering time. In a sample of 112 genotypes collected throughout the European range of the species, both seed dormancy and flowering time follow a latitudinal gradient independent of the major population structure gradient. This finding confirms previous studies reporting the adaptive evolution of these two traits. Here, however, we further analyze patterns of co-variation among traits. We observe that co-variation between primary dormancy, vegetative growth rate and flowering time also follows a latitudinal cline. At higher latitudes, vegetative growth rate is positively correlated with primary dormancy and negatively with flowering time. In the South, this trend disappears. Patterns of trait co-variation change, presumably because major environmental gradients shift with latitude. This pattern appears unrelated to population structure, suggesting that changes in the coordinated evolution of major life history traits is adaptive. Our data suggest that A. thaliana provides a good model for the evolution of trade-offs and their genetic basis

Understanding the genetic bases of natural variation for developmental and stress-related traits is a major goal of current plant biology. Variation in plant hormone levels and signaling might underlie such phenotypic variation occurring even within the same species. Here we report the genetic and molecular basis of semidwarf individuals found in natural Arabidopsis thaliana populations. Allelism tests demonstrate that independent loss-offunction mutations at GA locus 5 (GA5), which encodes gibberellin 20-oxidase 1 (GA20ox1) involved in the last steps of gibberellin biosynthesis, are found in different populations from southern, western, and northern Europe; central Asia; and Japan. Sequencing of GA5 identified 21 different loss-of-function alleles causing semidwarfness without any obvious general tradeoff affecting plant performance traits. GA5 shows signatures of purifying selection, whereas GA5 loss-of-function alleles can also exhibit patterns of positive selection in specific populations as shown by Fay and Wu’s H statistics. These results suggest that antagonistic pleiotropy might underlie the occurrence of GA5 loss-of-function mutations in nature. Furthermore, because GA5 is the ortholog of rice SD1 and barley Sdw1/Denso green revolution genes, this study illustrates the occurrence of conserved adaptive evolution between wild A.thaliana and domesticated plants

Knowledge of the exact distribution of meiotic crossovers (COs) and gene conversions (GCs) is essential for understanding many aspects of population genetics and evolution, from haplotype structure and long-distance genetic linkage to the generation of new allelic variants of genes. To this end, we resequenced the four products of 13 meiotic tetrads along with 10 doubled haploids derived from Arabidopsis thaliana hybrids. GC detection through short reads has previously been confounded by genomic rearrangements. Rigid filtering for misaligned reads allowed GC identification at high accuracy and revealed an ~80-kb transposition, which undergoes copy-number changes mediated by meiotic recombination. Non-crossover associated GCs were extremely rare most likely due to their short average length of ~25-50 bp, which is significantly shorter than the length of CO-associated GCs. Overall, recombination preferentially targeted non-methylated nucleosome-free regions at gene promoters, which showed significant enrichment of two sequence motifs.

A recombinant inbred line (RIL) population was produced based on a wide cross between the rapid-cycling and self-compatible genotypes L58, a Caixin vegetable type, and R-o-18, a yellow sarson oil type. A linkage map based on 160 F7 lines was constructed using 100 Single nucleotide polymorphisms (SNPs), 130 AFLP®, 27 InDel, and 13 publicly available SSR markers. The map covers a total length of 1150 centiMorgan (cM) with an average resolution of 4.3 cM/marker. To demonstrate the versatility of this new population, 17 traits, related to plant architecture and seed characteristics, were subjected to quantitative trait loci (QTL) analysis. A total of 47 QTLs were detected, each explaining between 6 and 54% of the total phenotypic variance for the concerned trait. The genetic analysis shows that this population is a useful new tool for analyzing genetic variation for interesting traits in B. rapa, and for further exploitation of the recent availability of the B. rapa whole genome sequence for gene cloning and gene function analysis.

Most plant species rely on seeds for their dispersal and survival under unfavorable environmental conditions. Seeds are characterized by their low moisture content and significantly reduced metabolic activities. During the maturation phase, seeds accumulate storage reserves and become desiccation-tolerant and dormant. Growth is resumed after release of dormancy and the occurrence of favorable environmental conditions. Here we show that embryonic cotyledon nuclei of Arabidopsis thaliana seeds have a significantly reduced nuclear size, which is established at the beginning of seed maturation. In addition, the chromatin of embryonic cotyledon nuclei from mature seeds is highly condensed. Nuclei regain their size and chromatin condensation level during germination. The reduction in nuclear size is controlled by the seed maturation regulator ABSCISIC ACID-INSENSITIVE 3, and the increase during germination requires two predicted nuclear matrix proteins, LITTLE NUCLEI 1 and LITTLE NUCLEI 2. Our results suggest that the specific properties of nuclei in ripe seeds are an adaptation to desiccation, independent of dormancy. We conclude that the changes in nuclear size and chromatin condensation in seeds are independent, developmentally controlled processes

To exploit the diversity in Arabidopsis thaliana, eight founder accessions were crossed to produce six recombinant inbred line (RIL) subpopulations, together called an Arabidopsis multiparent RIL (AMPRIL) population. Founders were crossed pairwise to produce four F1 hybrids. These F1s were crossed according to a diallel scheme. The resulting offspring was then selfed for three generations. The F4 generation was genotyped with SNP and microsatellite markers. Data for flowering time and leaf morphology traits were determined in the F5 generation. Quantitative trait locus (QTL) analysis for these traits was performed using especially developed mixed-model methodology, allowing tests for QTL main effects, QTL by background interactions, and QTL by QTL interactions. Because RILs were genotyped in the F4 generation and phenotyped in the F5 generation, residual heterozygosity could be used to confirm and fine-map a number of the QTLs in the selfed progeny of lines containing such heterozygosity. The AMPRIL population is an attractive resource for the study of complex traits.

A Quantitative Trait Locus (QTL) analysis was performed using two novel Recombinant Inbred Line (RIL) populations, derived from the progeny between two Arabidopsis thaliana genotypes collected at the same site in Kyoto (Japan) crossed with the reference laboratory strain Landsberg erecta (Ler). We used these two RIL populations to determine the genetic basis of seed dormancy and flowering time, which are assumed to be the main traits controlling life history variation in Arabidopsis. The analysis revealed quantitative variation for seed dormancy that is associated with allelic variation at the seed dormancy QTL DOG1 (for Delay Of Germination 1) in one population and at DOG6 in both. These DOG QTL have been previously identified using mapping populations derived from accessions collected at different sites around the world. Genetic variation within a population may enhance its ability to respond accurately to variation within and between seasons. In contrast, variation for flowering time, which also segregated within each mapping population, is mainly governed by the same QTL

Timing of germination is presumably under strong natural selection as it determines the environmental conditions in which a plant germinates and initiates its postembryonic life cycle. To investigate how seed dormancy is controlled, quantitative trait loci (QTL) analyses has been performed in six Arabidopsis thaliana recombinant inbred line populations by analyzing them simultaneously using a mixed model QTL approach. The recombinant inbred line populations were derived from crosses between the reference accession Landsberg erecta (Ler) and accessions from different world regions. In total, 11 delay of germination (DOG) QTL have been identified, and nine of them have been confirmed by near isogenic lines (NILs). The absence of strong epistatic interactions between the different DOG loci suggests that they affect dormancy mainly by distinct genetic pathways. This was confirmed by analyzing the transcriptome of freshly harvested dry seeds of five different DOG NILs. All five DOG NILs showed discernible and different expression patterns compared with the expression of their genetic background Ler. The genes identified in the different DOG NILs represent largely different gene ontology profiles. It is proposed that natural variation for seed dormancy in Arabidopsis is mainly controlled by different additive genetic and molecular pathways rather than epistatic interactions, indicating the involvement of several independent pathways

Chinese cabbage (Brassica rapa L. ssp. pekinensis) is one of the most important vegetables in China. Genetic dissection of leaf mineral accumulation and tolerance to Zn stress is important for the improvement of the nutritional quality of Chinese cabbage by breeding. A mapping population with 183 doubled haploid (DH) lines was used to study the genetics of mineral accumulation and the growth response to Zn. The genetic map was constructed based on 203 AFLPs, 58 SSRs, 22 SRAPs and four ESTPs. The concentration of 11 minerals was determined in leaves for 142 DH lines grown in an open field. In addition shoot dry biomass (SDB) under normal, deficient and excessive Zn nutritional conditions were investigated in hydroponics experiments. Ten QTLs, each explaining 11.1¿17.1% of the Na, Mg, P, Al, Fe, Mn, Zn and Sr concentration variance, were identified by multiple-QTL model (MQM) mapping. One common QTL was found affecting SDB under normal, deficient and excessive Zn nutritional conditions. An additional QTL was detected for SDB under Zn excess stress only. These results offer insights into the genetic basis of leaf mineral accumulation and plant growth under Zn stress conditions in Chinese cabbage

To improve QTL detection power for QTL main effects and interactions and QTL mapping resolution, new types of multi-founder crossing populations are created in plants and animals. Some recent examples are complex intercrossed populations in mice and Arabidopsis thaliana. For the latter, a set of eight accessions was intercrossed to produce four two-way hybrids that were subsequently intercrossed again in a half diallel fashion leading to six subpopulations of four-way hybrids, each subpopulation containing 100 individuals. Within each subpopulation, individuals were inbred for four generations via single seed descent. QTL mapping in the complex crosses requires new statistical tools. We present a first sketch of a QTL mapping methodology for the complex cross in Arabidopsis based on mixed model analyses. As experimental data were not yet available, we illustrate our methodology on simulated but realistic data.

Seed dormancy allows seeds to overcome periods that are unfavourable for seedling established and is therefore important for plant ecology and agriculture. Several processes are known to be involved in the induction of dormancy and in the switch from the dormant to the germinating state. The role of plant hormones, the different tissues and genes involved, including newly identified genes in dormancy and germination are described in this chapter, as well as the use transcriptome, proteome and metabolome analyses to study these mechanistically not well understood processes.

Background Plant primary carbohydrate metabolism is complex and flexible, and is regulated at many levels. Changes of transcript levels do not always lead to changes in enzyme activities, and these do not always affect metabolite levels and fluxes. To analyze interactions between these three levels of function, we have performed parallel genetic analyses of 15 enzyme activities involved in primary carbohydrate metabolism, transcript levels for their encoding structural genes, and a set of relevant metabolites. Quantitative analyses of each trait were performed in the Arabidopsis thaliana Ler × Cvi recombinant inbred line (RIL) population and subjected to correlation and quantitative trait locus (QTL) analysis. Results Traits affecting primary metabolism were often correlated, possibly due to developmental control affecting multiple genes, enzymes, or metabolites. Moreover, the activity QTLs of several enzymes co-localized with the expression QTLs (eQTLs) of their structural genes, or with metabolite accumulation QTLs of their substrates or products. In addition, many trait-specific QTLs were identified, revealing that there is also specific regulation of individual metabolic traits. Regulation of enzyme activities often occurred through multiple loci, involving both cis- and trans-acting transcriptional or post-transcriptional control of structural genes, as well as independently of the structural genes. Conclusion Future studies of the regulatory processes in primary carbohydrate metabolism will benefit from an integrative genetic analysis of gene transcription, enzyme activity, and metabolite content. The multiparallel QTL analyses of the various interconnected transducers of biological information flow, described here for the first time, can assist in determining the causes and consequences of genetic regulation at different levels of complex biological systems

Association mapping was used to investigate the genetic basis of variation within Brassica rapa, which is an important vegetable and oil crop. We analyzed the variation of phytate and phosphate levels in seeds and leaves and additional developmental and morphological traits in a set of diverse B. rapa accessions and tested association of these traits with AFLP markers. The analysis of population structure revealed four subgroups in the population. Trait values differed between these subgroups, thus defining associations between population structure and trait values, even for traits such as phytate and phosphate levels. Marker¿trait associations were investigated both with and without taking population structure into account. One hundred and seventy markers were found to be associated with the observed traits without correction for population structure. Association analysis with correction for population structure led to the identification of 27 markers, 6 of which had known map positions; 3 of these were confirmed in additional QTL mapping studies

In the future, more food needs to be produced with increasingly scarce natural resources. Genomics can play a key role in accelerating yield gains because it helps to improve our understanding ofgenetic traits and assists in breeding for better crop performance. The scientific muscle of genomicsattracted tremendous research investments, but the efficiency with which these investments are paying offis still low. How can we accelerate the application of molecular genetics to our understanding of crop physiology and subsequently to crop improvement? The missing link is a more detailed understanding of the effects of gene function on crop performance at field level under agronomically relevant conditions captured in robust, physiology-based mechanistic models. With such models the most sensitive processes and mechanisms at whole-crop level that contribute to improved crop performance can be identified. To achieve the detailed understanding necessary to build and feed these models, more research on wholeplant physiology and crop ecology is required, with a focus on the complexity of scaling up knowledge from the molecular level to the farmers¿ fields and production systems. Such studies assess how the plant is able to integrate the information at different levels of organization into the functioning of the whole plant and predicting the phenotype of transgenic plants engineered for improvement of a complex trait. More investment is needed in linking whole-plant physiology, crop ecology and crop simulation with molecular biology and genomics. Moreover, long-term progress can be enhanced by the formation of multidisciplinary teams that operate through networks of excellence in developing quantitative tools that integrate complex information and different levels of organization and by the exchange of young scientists between research groups working at different hierarchical levels. On the short term improvement of the characterization of experimental environments (preferabl

Accessions of a plant species can show considerable genetic differences that are analyzed effectively by using recombinant inbred line (RIL) populations. Here we describe the results of genome-wide expression variation analysis in an RIL population of Arabidopsis thaliana. For many genes, variation in expression could be explained by expression quantitative trait loci (eQTLs). The nature and consequences of this variation are discussed based on additional genetic parameters, such as heritability and transgression and by examining the genomic position of eQTLs versus gene position, polymorphism frequency, and gene ontology. Furthermore, we developed an approach for genetic regulatory network construction by combining eQTL mapping and regulator candidate gene selection. The power of our method was shown in a case study of genes associated with flowering time, a well studied regulatory network in Arabidopsis. Results that revealed clusters of coregulated genes and their most likely regulators were in agreement with published data, and unknown relationships could be predicted.

Brassica rapa L. is an important vegetable crop in eastern Asia. The objective of this study was to investigate the genetic variation in leaf Zn, Fe and Mn accumulation, Zn toxicity tolerance and Zn efficiency in B. rapa. In total 188 accessions were screened for their Zn-related characteristics in hydroponic culture. In experiment 1, mineral assays on 111 accessions grown under sufficient Zn supply (2 ¿M ZnSO4) revealed a variation range of 23.2-155.9 ¿g g¿1 dry weight (d. wt.) for Zn, 60.3-350.1 ¿g g¿1 d. wt. for Fe and 20.9-53.3 ¿g g¿1 d. wt. for the Mn concentration in shoot. The investigation of tolerance to excessive Zn (800 ¿M ZnSO4) on 158 accessions, by using visual toxicity symptom parameters (TSPs), identified different levels of tolerance in B. rapa. In experiment 2, a selected sub-set of accessions from experiment 1 was characterized in more detail for their mineral accumulation and tolerance to excessive Zn supply (100 ¿M and 300 ¿M ZnSO4). In this experiment Zn tolerance (ZT) determined by relative root or shoot dry biomass varied about 2-fold. The same six accessions were also examined for Zn efficiency, determined as relative growth under 0 ¿M ZnSO4 compared to 2 ¿M ZnSO4. Zn efficiency varied 1.8-fold based on shoot dry biomass and 2.6-fold variation based on root dry biomass. Zn accumulation was strongly correlated with Mn and Fe accumulation both under sufficient and deficient Zn supply. In conclusion, there is substantial variation for Zn accumulation, Zn toxicity tolerance and Zn efficiency in Brassica rapa L., which would allow selective breeding for these traits.

The micronutrient zinc has an essential role in physiological and metabolic processes in plants as a cofactor or structural element in 300 catalytic and noncatalytic proteins, but it is very toxic when available in elevated amounts. Plants tightly regulate their internal zinc concentrations in a process called zinc homeostasis. The exceptional zinc hyperaccumulator species Thlaspi caerulescens can accumulate up to 3% of zinc, but also high amounts of nickel and cadmium, without any sign of toxicity. This should have drastic effects on the zinc homeostasis mechanism. We examined in detail the transcription profiles of roots of Arabidopsis thaliana and T. caerulescens plants grown under deficient, sufficient, and excess supply of zinc. A total of 608 zinc-responsive genes with at least a 3-fold difference in expression level were detected in A. thaliana and 352 in T. caerulescens in response to changes in zinc supply. Only 14% of these genes were also zinc responsive in A. thaliana. When comparing A. thaliana with T. caerulescens at each zinc exposure, more than 2,200 genes were significantly differentially expressed (≥5-fold and false discovery rate < 0.05). While a large fraction of these genes are of yet unknown function, many genes with a different expression between A. thaliana and T. caerulescens appear to function in metal homeostasis, in abiotic stress response, and in lignin biosynthesis. The high expression of lignin biosynthesis genes corresponds to the deposition of lignin in the endodermis, of which there are two layers in T. caerulescens roots and only one in A. thaliana. © 2006 American Society of Plant Biologists.

The micronutrient zinc has an essential role in physiological and metabolic processes in plants as a cofactor or structural element in 300 catalytic and noncatalytic proteins, but it is very toxic when available in elevated amounts. Plants tightly regulate their internal zinc concentrations in a process called zinc homeostasis. The exceptional zinc hyperaccumulator species Thlaspi caerulescens can accumulate up to 3% of zinc, but also high amounts of nickel and cadmium, without any sign of toxicity. This should have drastic effects on the zinc homeostasis mechanism. We examined in detail the transcription profiles of roots of Arabidopsis thaliana and T. caerulescens plants grown under deficient, sufficient, and excess supply of zinc. A total of 608 zinc-responsive genes with at least a 3-fold difference in expression level were detected in A. thaliana and 352 in T. caerulescens in response to changes in zinc supply. Only 14% of these genes were also zinc responsive in A. thaliana. When comparing A. thaliana with T. caerulescens at each zinc exposure, more than 2,200 genes were significantly differentially expressed (≥5-fold and false discovery rate < 0.05). While a large fraction of these genes are of yet unknown function, many genes with a different expression between A. thaliana and T. caerulescens appear to function in metal homeostasis, in abiotic stress response, and in lignin biosynthesis. The high expression of lignin biosynthesis genes corresponds to the deposition of lignin in the endodermis, of which there are two layers in T. caerulescens roots and only one in A. thaliana. © 2006 American Society of Plant Biologists.

Vitamin E is an essential nutrient for humans and is obtained primarily from food, especially oil, derived from the seed of plants. Genes encoding the committed steps in vitamin E synthesis in plants (VTE, loci 1-5) have been isolated and used for tocopherol pathway engineering with various degrees of success. As a complement to such approaches we have used quantitative trait loci analysis with two sets of Arabidopsis thaliana recombinant inbred lines and have identified 14 QVE (quantitative vitamin E) loci affecting tocopherol content and composition in seeds. Five QVE intervals contain VTE loci that are likely QVE gene candidates. Nine QVE intervals do not contain VTE loci and therefore identify novel loci affecting seed tocopherol content and composition. Several near-isogenic lines containing introgressions of the accession with increased vitamin E levels were shown to confer significantly elevated tocopherol levels compared with the recurrent parent. Fine-mapping has narrowed QVE7 (a gamma-tocopherol quantitative trait loci) to an 8.5-kb interval encompassing two genes. Understanding the basis of the QVE loci in Arabidopsis promises to provide insight into the regulation and/or metabolism of vitamin E in plants and has clear ramifications for improving the nutritional content of crops through marker-assisted selection and metabolic engineering.

The possible role of the sucrose-splitting enzymes sucrose synthase and invertase in elongating roots and hypocotyls of Arabidopsis was tested by using a combination of histochemical methods and quantitative trait locus (QTL) analysis. Lengths of roots and hypocotyls correlated better with invertase activities than with sucrose synthase activities. The highest correlations were observed with activities in the elongating zones of roots. The genetic basis of these correlations was studied by using QTL analysis. Several loci, affecting invertase activity, colocated with loci that had an effect on root or hypocotyl length. Further fine mapping of a major locus for root length, but not for hypocotyl length (top chromosome 1), consistently showed colocation with the locus for invertase activity containing a gene coding for a vacuolar invertase. The analysis of a functional knockout line confirmed the role of this invertase in root elongation, whereas other invertase genes might play a role in hypocotyl elongation. Thus, we show the power of QTL analysis, combined for morphological and biochemical traits, followed by fine-mapping and mutant analysis, in unraveling the function of genes and their role in growth and development

Genetic variation for seed dormancy in nature is a typical quantitative trait controlled by multiple loci on which environmental factors have a strong effect. Finding the genes underlying dormancy quantitative trait loci is a major scientific challenge, which also has relevance for agriculture and ecology. In this study we describe the identification of the DELAY OF GERMINATION 1 (DOG1) gene previously identified as a quantitative trait locus involved in the control of seed dormancy. This gene was isolated by a combination of positional cloning and mutant analysis and is absolutely required for the induction of seed dormancy. DOG1 is a member of a small gene family of unknown molecular function, with five members in Arabidopsis. The functional natural allelic variation present in Arabidopsis is caused by polymorphisms in the cis-regulatory region of the DOG1 gene and results in considerable expression differences between the DOG1 alleles of the accessions analyzed

Natural variation for freezing tolerance is a major component of adaptation and geographic distribution of plant species. However, little is known about the genes and molecular mechanisms that determine its naturally occurring diversity. We have analyzed the intraspecific freezing tolerance variation existent between two geographically distant accessions of Arabidopsis (Arabidopsis thaliana), Cape Verde Islands (Cvi) and Landsberg erecta (Ler). They differed in their freezing tolerance before and after cold acclimation, as well as in the cold acclimation response in relation to photoperiod conditions. Using a quantitative genetic approach, we found that freezing tolerance differences after cold acclimation were determined by seven quantitative trait loci (QTL), named FREEZING TOLERANCE QTL 1 (FTQ1) to FTQ7. FTQ4 was the QTL with the largest effect detected in two photoperiod conditions, while five other FTQ loci behaved as photoperiod dependent. FTQ4 colocated with the tandem repeated genes C-REPEAT BINDING FACTOR 1 (CBF1), CBF2, and CBF3, which encode transcriptional activators involved in the cold acclimation response. The low freezing tolerance of FTQ4-Cvi alleles was associated with a deletion of the promoter region of Cvi CBF2, and with low RNA expression of CBF2 and of several CBF target genes. Genetic complementation of FTQ4-Cvi plants with a CBF2-Ler transgene suggests that such CBF2 allelic variation is the cause of CBF2 misexpression and the molecular basis of FTQ4.

The model plant Arabidopsis thaliana has a broad natural distribution throughout the Northern Hemisphere therefor many different accessions could be collected from natural populations and are available for experimental analysis. These accessions provide an alternative source of genetic variation that can be used to study the function of genes.The main aim of the work presented in this thesis is to increase the knowledge about the genetic control of seed dormancy. Therefore the natural variation present between two Arabidopsis accessions; Landsberg erecta and Cape Verde Islands has been analysed for this character. This analyses revealed several loci involved in seed dormancy, one of them has been further characterised and fine-mapped. In addition other seed traits that might be related to seed dormancy and germination have been analysed. These traits are seed oligosaccharide content, seed storability and seed phytate and phosphate content.

Arabidopsis possesses dormancy, as is the case for many other plant species, which is controlled by environmental factors such as light, temperature and time of dry storage as well as by genetic factors. The use of genetics and molecular genetics in Arabidopsis is starting to shed light on some aspects of the mechanism of dormancy and germination by the identification of mutants and genes that control these processes. This review provides an overview of current knowledge of factors and genes controlling seed dormancy and germination in Arabidopsis

The natural allelic variation at flowering time loci between two Arabidopsis thaliana accessions; Landsberg erecta (L er ) and the tropical Cape Verde Island (Cvi) has been analyzed. Quantitative trait locus (QTL) mapping in recombinant inbred lines (RILs) grown under three environments, differing in day-length and/or vernalization treatment, was used to detect and locate flowering loci on the genetic map of Arabidopsis. Four main QTLs were identified, designated EDI , FLF , FLG and FLH , to which most of the flowering time differences could be attributed. At the EDI and FLH loci, Cvi alleles resulted in earliness, whereas at FLF and FLG , Cvi alleles caused lateness. Among these QTL, EDI ( EARLY, DAY-LENGTH INSENSITIVE ) is the main locus responsible for day-length sensitivity. To determine the molecular basis of this natural genetic variation, we have isolated the EDI locus using a map-based cloning strategy. The positional cloning of the EDI QTL showed it to be a novel allele of CRY2 , encoding the blue-light photoreceptor cryptochrome-2, that has previously been shown to promote flowering in long-day photoperiods. It was shown that the unique flowering phenotype in the CRY2 -Cvi allele results from a single amino acid substitution that reduces the down-regulation of the CRY2 protein in plants grown under short photoperiods, leading to early flowering. The genetic interaction between CRY2 and other loci involved in the different flowering promotion pathways was studied. The three different types of CRY2 alleles ( CRY2 -Cvi = EDI , CRY2 -L er , and the CRY2 null mutant fha1-1 ) were combined with the mutants representing the different flowering pathways. This analysis indicated that the so called "vernalization and autonomous pathway" that control FLC ( Flowering locus C ) expression, repress the expression of CRY2 .

For a long time, mutagenesis research in plants focused on crop improvement and, especially for crop plants, opimised protocols were developed with barley being one of the favourite species. However, the interest in mutagenesis has shifted to basic plant research in the last 20 years, when the power of mutant approaches in combination with molecular techniques to investigate the molecular nature of the genes became fully appreciated

Arabidopsis possesses dormancy, as is the case for many other plant species, which is controlled by environmental factors such as light, temperature and time of dry storage as well as by genetic factors. The use of genetics and molecular genetics in Arabidopsis is starting to shed light on some aspects of the mechanism of dormancy and germination by the identification of mutants and genes that control these processes. This review provides an overview of current knowledge of factors and genes controlling seed dormancy and germination in Arabidopsis

Arabidopsis thaliana has become the model for plant genetics and plant development because of its genetic characteristics, such as short generation time, the small size of its genome and the availability of its complete DNA sequence.

This thesis presents a diversity analysis for 26 tetraploid wheat landraces from the central region of Ethiopia and four cultivars using five markers: morphological traits, chromosome portraits, microsatellites, AFLP and storage proteins.The aim of the study is:To estimate the magnitude of genetic diversity within and between 26 landracesTo identify the pattern of genetic diversity within and between geographical areasTo detect duplications within and between accessionsTo prioritise markers used for estimating genetic diversity in Ethiopian tetraploid wheat landracesThe study revealed a great difference of diversity between the landraces and also demonstrated that the high variation is due to the within component of accessions rather than the between landrace component. No strong variation for genetic composition was observed between plants from different altitudes and subregions. An integrated application of morphological, chromosomal, molecular and storage protein markers for assessing biodiversity in these wheat landraces is strongly recommended.&nbsp

The continuous growth of the plant embryo is interrupted during the seed maturation processes which results in a dormant seed. The embryo continues development after germination when it grows into a seedling. The embryo growth phase starts after morphogenesis and ends when the embryo fills the seed sac. Very little is known about the processes regulating this phase. We describe mutants that affect embryo growth in two sequential developmental stages. Firstly, embryo growth arrest is regulated by the FUS3/LEC type genes, as mutations in these genes cause a continuation of growth in immature embryos. Secondly, a later stage of embryo dormancy is regulated by ABI3 and abscisic acid; abi3 and aba1 mutants exhibit premature germination only after embryos mature. Mutations affecting both developmental stages result in an additive phenotype and double mutants are highly viviparous. Embryo growth arrest is regulated by cell division activities in both the embryo and the endosperm, which are gradually switched off at the mature embryo stage. In the fus3/lec mutants, however, cell division in both the embryo and endosperm is not arrested, but rather is prolonged throughout seed maturation. Furthermore ectopic cell division occurs in seedlings. Our results indicate that seed dormancy is secured via at least two sequential developmental processes: embryo growth arrest, which is regulated by cell division and embryo dormancy.

The transition from the vegetative to the reproductive phase (flowering initiation) in plants has a complex regulation which is affected by environmental and internal plant factors. The understanding of this process is not only of fundamental interest but could also lead to practical applications. The research into flowering initiation has a long history. The initial emphasis on physiological and biochemical studies led to the identification of different factors that influence flowering time. During the sixties, a genetic approach was initiated in different plant species. In Arabidopsis several late flowering mutants were isolated and genetically and physiologically characterised which revealed a complex regulation of flowering time by different pathways. These are the photoperiodic promotion pathway which promotes flowering under long day light conditions, the vernalisation promotion pathway which promotes flowering by low temperatures and the autonomous promotion pathway which promotes flowering independent of the environment. Due to its favourable genetic and molecular features, research on flowering initiation became focussed on Arabidopsis. Since the beginning of the nineties, several of the genes involved in the different pathways have been cloned, providing more information about the function of these genes in the cell and their relations with each other. Despite this increasing amount of information, the picture is still far from complete.The aim of the work presented in this thesis is to increase our knowledge of flowering time regulation. It focussed on the genetic and molecular characterisation of the semi-dominant mutant fwa , which flowers late under long day light conditions and has been proposed to be part of the photoperiodic promotion pathway. One approach sought to identify additional genes that affect flowering by mutagenesis of the fwa mutant. In addition to three different intragenic revertants of fwa , this screen yielded a novel early flowerin

Abscisic acid (ABA) is a plant hormone involved in seed development and germination and in responses to various environmental stresses. The last step of ABA biosynthesis involves oxidation of abscisic aldehyde, and aldehyde oxidase (EC 1.2.3.1) is thought to catalyze this reaction. An aldehyde oxidase isoform, AO, encoded by AAO3, one of four Arabidopsis aldehyde oxidase genes (AAO1, AAO2, AAO3, and AAO4), is the most likely candidate for the enzyme, because it can efficiently catalyze the oxidation of abscisic aldehyde to ABA. Here, we report the isolation and characterization of an ABA-deficient Arabidopsis mutant that maps at the AAO3 locus. The mutant exhibits a wilty phenotype in rosette leaves, but seed dormancy is not affected. ABA levels were significantly reduced in the mutant leaves, explaining the wilty phenotype in rosettes, whereas the level in the mutant seeds was less reduced. No AO activity could be detected in the rosette leaves of the mutant. Sequence data showed that the mutant contains a G to A substitution in the AAO3 gene. The mutation causes incorrect splicing of the ninth intron of AAO3 mRNA. We thus conclude that the ABA-deficient mutant is impaired in the AAO3 gene and that the gene product, AO, is an aldehyde oxidase that catalyzes the last step of ABA biosynthesis in Arabidopsis, specifically in rosette leaves. Other aldehyde oxidases may be involved in ABA biosynthesis in other organs.