Association mapping of cadmium, copper and hydrogen peroxide tolerance of roots and translocation capacities of cadmium and copper in Arabidopsis thaliana

Association mapping analysis of Cd, Cu and H ₂O ₂ tolerance, judged by relative root length (RRL: % of root length in stress condition relative to that in control condition), and Cd and Cu translocation ratios (amount of metal in the shoot to the total) were performed using 90 accessions of Arabidopsis thaliana . Using 140 SNPs that were distributed across the genome, association mapping analysis was performed with a haploid setting by the Q + K method, which minimizes detection of false associations by combining the Q-matrix of the structured association (Q) with kinship (K) to control for the population structure. Six, five and five significant (−log ₁₀ P -value is 1.3 ≥) linkages were detected between the SNPs and Cd, Cu and H₂O₂ resistant RRLs, respectively. In addition, six significant linkages were identified with translocation capacities of Cd and Cu. Among those detected loci, two each of Cu and Cd tolerance RRLs were collocated with those of H₂O₂ tolerance RRL, while one locus each was detected by Cu and Cd tolerance RRLs that collocated with their translocation ratios. These results suggested that these factors might partly explain the phenotypic variation of tolerance RRLs to Cd and Cu of Arabidopsis thaliana . Finally, using a different approach to analyze interactions between individual phenotypes, namely clustering analysis, we found an expected segregation of resistant SNPs (single-nucleotide polymorphisms) of the multiple RRLs in the typical accession groups carrying multiple traits. Almost none of the loci detected by association mapping analysis were linked to the loci of previously identified critical genes regulating the traits, suggesting that this could be useful to identify complex architecture of genetic factors determining variation among multiple accessions.     

Natural variation of Arabidopsis response to nitrogen availability

Our understanding of plant growth in response to nitrogen (N) supply is mainly based on studies of mutants and transformants. This study explored the natural variability of Arabidopsis thaliana first to find out its global response to N availability and secondly to characterize the plasticity for growth and N metabolism among 23 genetically distant accessions under normal (N+), limited (N-), and starved (N0) N supplies. Plant growth was estimated by eight morphological traits characterizing shoot and root growth and 10 metabolic parameters that represented N and carbon metabolism. Most of the studied traits showed a large variation linked to genotype and nutrition. Furthermore, Arabidopsis growth was coordinated by master traits such as the shoot to root ratio of nitrate content in N+, root fresh matter and root amino acids in N-, and shoot fresh matter together with root thickness in N0. The 23 accessions could be gathered into four different groups, according to their growth in N+, N-, and N0. Phenotypic profiling characterized four different adaptative responses to N- and N0. Class 1 tolerated N limitation with the smallest decrease in shoot and root biomass compared with N+, while class 2 presented the highest resistance to N starvation by preferential increased root growth, huge starch accumulation, and high shoot nitrate content. In contrast, class 3 plants could tolerate neither N limitation nor N starvation. Small plants of class 4 were different, with shoot biomass barely affected in N- and root biomass unaffected in N0.     

Characterization of natural variation for zinc, iron and manganese accumulation and zinc exposure response in Brassica rapa L.

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 μM ZnSO₄) revealed a variation range of 23.2–155.9 μg g⁻¹ dry weight (d. wt.) for Zn, 60.3–350.1 μg g⁻¹ d. wt. for Fe and 20.9–53.3 μg g⁻¹ d. wt. for the Mn concentration in shoot. The investigation of tolerance to excessive Zn (800 μM 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 μM and 300 μM 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 μM ZnSO₄ compared to 2 μM 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.     

Natural variation of submergence tolerance among Arabidopsis thaliana accessions

? The exploitation of natural variation in Arabidopsis thaliana (Arabidopsis) provides a huge potential for the identification of the molecular mechanisms underlying this variation as a result of the availability of a vast array of genetic and genomic resources for this species. Eighty-six Arabidopsis accessions were screened for natural variation in flooding tolerance. This forms the first step towards the identification and characterization of the role of candidate genes contributing to flooding tolerance. ? Arabidopsis accessions at the 10-leaf stage were subjected to complete submergence in the dark. Survival curves were plotted to estimate median lethal times as a measure of tolerance. Flooding-associated survival parameters, such as root and shoot oxygen content, initial carbohydrate content and petiole elongation under water, were also measured. ? There was a significant variation in submergence tolerance among Arabidopsis accessions. However, the order of tolerance did not correlate with root and shoot oxygen content or initial amounts of shoot starch and total soluble sugars. A negative correlation was observed between submergence tolerance and underwater petiole elongation. ? Arabidopsis accessions show considerable variation in the ability to tolerate complete submergence, making it a good species in which to identify and characterize genes and to study mechanisms that contribute to survival under water.     

Variation in root-to-shoot translocation of cadmium and zinc among different accessions of the hyperaccumulators Thlaspi caerulescens and Thlaspi praecox

Efficient root-to-shoot translocation is a key trait of the zinc/cadmium hyperaccumulators Thlaspi caerulescens and Thlaspi praecox, but the extent of variation among different accessions and the underlying mechanisms remain unclear. Root-to-shoot translocation of Cd and Zn and apoplastic bypass flow were determined in 10 accessions of T. caerulescens and one of T. praecox, using radiolabels (109)Cd and (65)Zn. Two contrasting accessions (Pr and Ga) of T. caerulescens were further characterized for TcHMA4 expression and metal compartmentation in roots. Root-to-shoot translocation of (109)Cd and (65)Zn after 1 d exposure varied 4.4 to 5-fold among the 11 accessions, with a significant correlation between the two metals, but no significant correlation with uptake or the apoplastic bypass flow. The F(2) progeny from a cross between accessions from Prayon, Belgium (Pr) and Ganges, France (Ga) showed a continuous phenotype pattern and transgression. There was no significant difference in the TcHMA4 expression in roots between Pr and Ga. Compartmentation analysis showed a higher percentage of (109)Cd sequestered in the root vacuoles of Ga than Pr, the former being less efficient in translocation than the latter. Substantial natural variation exists in the root-to-shoot translocation of Cd and Zn, and root vacuolar sequestration may be an important factor related to this variation.     

Variation in selenium tolerance and accumulation among 19 Arabidopsis thaliana accessions

Selenium (Se) is an essential element for many organisms but also toxic at higher levels. The objective of this study was to identify accessions from the model species Arabidopsis thaliana that differ in Se tolerance and accumulation. Nineteen Arabidopsis accessions were grown from seed on agar medium with or without selenate (50 microM) or selenite (20 microM), followed by analysis of Se tolerance and accumulation. Tissue sulfur levels were also compared. The Se Tolerance Index (root length+Se/root length control) varied among the accessions from 0.11 to 0.44 for selenite and from 0.05 to 0.24 for selenate. When treated with selenite, the accessions differed by two-fold in shoot Se concentration (up to 250 mgkg(-1)) and three-fold in root Se concentration (up to 1000 mgkg(-1)). Selenium accumulation from selenate varied 1.7-fold in shoot (up to 1000 mgkg(-1)) and two-fold in root (up to 650 mgkg(-1)). Across all accessions, a strong correlation was observed between Se and S concentration in both shoot and root under selenate treatment, and in roots of selenite-treated plants. Shoot Se accumulation from selenate and selenite were also correlated. There was no correlation between Se tolerance and accumulation, either for selenate or selenite. The F(1) offspring from a cross between the extreme selenate-sensitive Dijon G and the extreme selenate-tolerant Estland accessions showed intermediate selenate tolerance. In contrast, the F(1) offspring from a cross between selenite-sensitive and -tolerant accessions (Dijon GxCol-PRL) were selenite tolerant. The results from this study give new insight into the mechanisms of plant selenium (Se) tolerance and accumulation, which may help develop better plants for selenium phytoremediation or as fortified foods.     

Effects of suboptimal root zone temperatures and shoot demand on net translocation of micronutrients from the roots to the shoot of maize

The effects of suboptimal root zone temperatures (RZTs) on net translocation rates from the roots to the shoots and the concentrations of Fe, Mn, Zn, and Cu were examined in maize grown in nutrient solution or soil. Plants were grown at 12 °C, 18 °C and 24 °C RZT. At each RZT, the growth-related shoot demand for nutrients was varied by independently modifying the temperature of the shoot base (SBT) including the apical shoot meristem. The net translocation rates of Mn and Zn from the roots to the shoots were reduced at low RZTs, irrespective of the SBT and of the substrate (soil or nutrient solution). Obviously, the net translocation rates of Mn and Zn at low RZT were mainly regulated by temperature effects on the roots and not by the chemical nutrient availability in the rhizosphere or by shoot growth rate as controlled by SBTs. When both RZT and SBT were reduced, the decrease in net translocation rates of Mn and Zn was similar to the decline in the shoot growth rate and concentrations of Mn and Zn in the shoot fresh matter were not greatly affected or were even increased by low RZT. However, at high SBT and low RZT in nutrient solution, the depressed net translocation rates of Mn and Zn combined with the increased shoot growth resulted in significantly decreased concentrations of Mn and Zn in the shoot, indicating that Mn and Zn may become deficient even at high chemical availability. By contrast to Mn and Zn, the net translocation rates of Fe and Cu at all RZTs were markedly enhanced by increased SBTs. Accordingly, the concentrations of Fe and Cu in the shoot fresh matter were not greatly affected by RZTs, irrespective of the SBTs. These results indicate that the ability of roots to supply Fe and Cu to the shoot was internally regulated by the growth related shoot demand per unit of roots. Deceased 21 September 1996 Deceased 21 September 1996     

MTP1-dependent Zn sequestration into shoot vacuoles suggests dual roles in Zn tolerance and accumulation in Zn-hyperaccumulating plants

The integral membrane protein Thlaspi goesingense metal tolerance protein 1 (TgMTP1) has been suggested to play an important role in Zn hyperaccumulation in T. goesingense . Here, we show that the TgMTP1 protein is accumulated to high levels at the vacuolar membrane in shoot tissue of T. goesingense . TgMTP1 is likely to act in the transport of Zn into the vacuole, enhancing both Zn accumulation and tolerance. By specifically expressing TgMTP1 in Arabidopsis thaliana shoots, we show that TgMTP1, localized at the vacuolar membrane, can drive the enhanced shoot accumulation of Zn by initiating a systemic Zn deficiency response. The systematic response includes increased expression of Zn transporters ( ZIP3 , ZIP4 , ZIP5 and ZIP9 ) in both shoot and root tissue. Furthermore, shoot-specific accumulation of TgMTP1 at the vacuolar membrane also leads to increased resistance to Zn in A. thaliana , probably through enhanced Zn compartmentalization in the vacuole. Such evidence leads to the conclusion that the high levels of TgMTP1 at the vacuolar membrane in shoot tissue of the Zn hyperaccumulator T. goesingense play a role in both Zn tolerance and enhanced Zn uptake and accumulation, via the activation of a systemic Zn deficiency response.     

The Arabidopsis metal tolerance protein AtMTP3 maintains metal homeostasis by mediating Zn exclusion from the shoot under Fe deficiency and Zn oversupply

Zinc ions are required to maintain the biological activity of numerous proteins. However, when mislocalized or accumulated in excess, Zn(2+) ions are toxic because of adventitious binding to proteins and displacement of other metal ions, among them Fe(2+), from their binding sites. Heterologous expression of a previously uncharacterized Arabidopsis thaliana metal tolerance protein, MTP3, in the zrc1 cot1 mutant of budding yeast restores tolerance to, and cellular accumulation of, zinc and cobalt. An MTP3-GFP fusion protein localizes to the vacuolar membrane when expressed in Arabidopsis. Ectopic over-expression of MTP3 increases Zn accumulation in both roots and rosette leaves of A. thaliana, and enhances Zn tolerance. Exposure of wild-type plants to high but non-toxic concentrations of Zn or Co, or Fe deficiency, strongly induce MTP3 expression specifically in epidermal and cortex cells of the root hair zone. Silencing of MTP3 by RNA interference causes Zn hypersensitivity and enhances Zn accumulation in above-ground organs of soil-grown plants and of seedlings exposed to excess Zn or to Fe deficiency. Our data indicate that, in wild-type A. thaliana, the AtMTP3 protein contributes to basic cellular Zn tolerance and controls Zn partitioning, particularly under conditions of high rates of Zn influx into the root symplasm.     

Waterlogging Tolerance Among a Diverse Range of Trifolium Accessions is Related to Root Porosity, Lateral Root Formation and 'Aerotropic Rooting'

Waterlogging tolerance, root porosity and root anatomy wereevaluated for 20 Trifolium accessions (species and sub-species,all annuals) selected from the eight Sections of the genus.Nine accessions were sensitive [relative growth rate (RGR) reducedby up to 80%] to waterlogging, nine accessions were tolerant(RGR not reduced), and in two accessions RGR increased (up to1.9-fold), when compared to drained controls. Growth of themain (i.e. tap) root axis was severely reduced in all accessionswhen waterlogged. Lateral roots formed the bulk of the rootsystem of tolerant accessions when grown in waterlogged soil.Lengths of the longest lateral roots were up to three-timeslonger than the main root axis. Root porosity varied from 0.7–12%among accessions when grown in aerated solution and from 1.1–15.5%in plants grown in hypoxic (0.031–0.045 mol O₂m^-3) solution.In some accessions aerenchyma formed by cell lysigeny; in othersit formed by schizogenous cell separation, or a combinationof both processes. O₂consumption rates of expanded lateral roottissues varied by up to 1.7-fold (on a mass basis) among thesix accessions tested and was reduced by an average of 24% forroots of plants grown in hypoxic solution prior to measurements.Accessions with the highest root porosity tended to have longerroots when grown in waterlogged soil. Three accessions formed‘aerotropic roots’ and the lateral root lengthsof these plants exceeded those of all other accessions, suggestingenhanced O₂movement to the submerged lateral root axis via theaerotropic roots. Waterlogging-tolerant accessions were identifiedin seven of the eight Sections in Trifolium, and the tolerantaccessions tended to be those with extensive lateral root systemsof relatively high porosity. Copyright 2001 Annals of BotanyCompany Waterlogging, Trifolium, aerenchyma, hypoxia, flooding, root respiration, clover, root anatomy, root porosity, pasture, aerotropic roots     

Overexpression of AtHMA4 enhances root-to-shoot translocation of zinc and cadmium and plant metal tolerance

AtHMA4 is an Arabidopsis thaliana P1B-ATPase which transports Zn and Cd. Here, we demonstrate that AtHMA4 is localized at the plasma membrane and expressed in tissues surrounding the root vascular vessels. The ectopic overexpression of AtHMA4 improved the root growth in the presence of toxic concentrations of Zn, Cd and Co. A null mutant exhibited a lower translocation of Zn and Cd from the roots to shoot. In contrast, the AtHMA4 overexpressing lines displayed an increase in the zinc and cadmium shoot content. Altogether, these results strongly indicate that AtHMA4 plays a role in metal loading in the xylem.     

Tolerance to Zinc Deficiency in Rice Correlates with Zinc Uptake and Translocation

To study variation in zinc efficiency (ZE) among current Chinese rice genotypes, a pot experiment was conducted with 15 aerobic and 8 lowland rice genotypes. Aerobic rice is currently bred by crossing lowland with upland rice genotypes, for growth in an aerobic cultivation system, which is saving water and producing high yields. A Zn deficient clay soil was used in our screening. Zn deficiency resulted in a marked decrease in shoot dry matter production of most genotypes after 28 days of growth. Genotypes were ranked according to their tolerance to Zn deficiency based on ZE, expressed as the ratio of shoot dry weight at Zn deficiency over that at adequate Zn supply. Substantial genotypic variation in ZE (50–98%) was found among both lowland and aerobic genotypes. ZE correlated significantly (P < 0.05) with Zn uptake (R ² = 0.34), Zn translocation from root to shoot (R ² = 0.19) and shoot Zn concentration (R ² = 0.27). The correlation with seed Zn content was insignificant. In stepwise multiple regression analyses, variation in Zn uptake and Zn translocation explained 53% of variation in ZE. Variation in Zn uptake could be explained only for 32% by root surface area. These results indicate that Zn uptake may be an important determinant of ZE and that mechanisms other than root surface area are of major importance in determining Zn uptake by rice.     

Variation in Cadmium Tolerance and Accumulation and Their Relationship in Wheat Recombinant Inbred Lines at Seedling Stage

In order to identify the variation of cadmium (Cd) tolerance and accumulation in wheat (Triticum aestivum L.), a study was conducted in hydroponic culture with or without Cd using recombinant inbred lines (RILs) consisting of 103 RILs derived from a cross of Chuan 35050 × Shannong 483 at seedling stage. The parameters of shoot height, secondary roots numbers, tiller numbers, shoot dry weights, root dry weights, and maximum efficiency of photosystem II photochemistry under dark-adopted conditions were measured. Cd-tolerant indexes were then calculated as relative the above traits under Cd stress to those under the control. Cd concentration in shoot or root was determined and Cd accumulation and translocation were calculated. Based on the Cd-tolerant indexes, membership function analysis was used to determine the variation of the above parameters. The results showed a continuous distribution among the RILs and then the RILs were divided into five groups according to their tolerance. Lines 76 and 17 were considered as the most Cd-tolerant lines while lines 103 and 51 were as the most Cd-sensitive lines. Meanwhile, lines 38 and 79 were with minimum Cd translocation ratio while lines 88 and 53 were with maximum Cd translocation ratio, respectively. The relationship between Cd tolerance and accumulation was not significant, indicating Cd tolerance and accumulation may be independent traits in the RILs. Thus, lines with high Cd tolerance and less Cd accumulation could be selected for wheat breeding.     

Aluminum exclusion from root zone and maintenance of nutrient uptake are principal mechanisms of Al tolerance in <Emphasis Type="Italic">Pisum sativum</Emphasis> L.

Our study aimed to evaluate intraspecific variability of pea (Pisum sativum L.) in Al tolerance and to reveal mechanisms underlying genotypic differences in this trait. At the first stage, 106 pea genotypes were screened for Al tolerance using root re-elongation assay based on staining with eriochrome cyanine R. The root re-elongation zone varied from 0.5 mm to 14 mm and relationships between Al tolerance and provenance or phenotypic traits of genotypes were found. Tolerance index (TI), calculated as a biomass ratio of Al-treated and non-treated contrasting genotypes grown in hydroponics for 10 days, varied from 30% to 92% for roots and from 38% to 90% for shoots. TI did not correlate with root or shoot Al content, but correlated positively with increasing pH and negatively with residual Al concentration in nutrient solution in the end of experiments. Root exudation of organic acid anions (mostly acetate, citrate, lactate, pyroglutamate, pyruvate and succinate) significantly increased in several Al-treated genotypes, but did not correlate with TI. Al-treatment decreased Ca, Co, Cu, K, Mg, Mn, Mo, Ni, S and Zn contents in roots and/or shoots, whereas contents of several elements (P, B, Fe and Mo in roots and B and Fe in shoots) increased, suggesting that Al toxicity induced substantial disturbances in uptake and translocation of nutrients. Nutritional disturbances were more pronounced in Al sensitive genotypes. In conclusion, pea has a high intraspecific variability in Al tolerance and this trait is associated with provenance and phenotypic properties of plants. Transformation of Al to unavailable (insoluble) forms in the root zone and the ability to maintain nutrient uptake are considered to be important mechanisms of Al tolerance in this plant species.     

The impact of limited soil moisture and waterlogging stress conditions on morphological and anatomical root traits in maize (Zea mays L.) hybrids of different drought tolerance

Effects of soil drought or waterlogging on the morphological traits of the root system and internal root anatomy were studied in maize hybrids of different drought tolerance. The investigations comprised quantitative and qualitative analyses of a developed plant root system through determining the number, length and dry matter of the particular components of the root system and some traits of the anatomical structure of the seminal root. Obtained results have demonstrated a relatively broad variation in the habit of the root system. This mainly refers, to the number, length and dry matter of lateral roots, developed by seminal root, seminal adventitious and nodal roots as well as to some anatomical properties of the stele, cortex and metaxylem elements. Plants grown under waterlogging or drought conditions showed a smaller number and less dry matter of lateral branching than plants grown in control conditions. The harmful effect of waterlogging conditions on the growth of roots was greater when compared with that of plants exposed to drought. In the measurements of the root morphological traits, the effect of soil drought on the internal root anatomical characteristic was weaker than the effect of soil waterlogging. The observed effects of both treatments were more distinct in a drought sensitive hybrid Pioneer D than in drought resistant Pioneer C one. The drought resistant hybrid Pioneer C distinguished by a more extensive rooting and by smaller alterations in the root morphology caused by the stress conditions than drought sensitive hybrid Pioneer D one. Also the differences between the resistant and the sensitive maize hybrids were apparent for examined root anatomical traits. Results confirm that the hybrid Pioneer D of a high drought susceptibility was found to be also more sensitive to periodieal soil water excess. A more efficient water use and a lower shoot to root (S:R) ratio were found to be major reasons for a higher stress resistance of the hybrid Pioneer C. The reasons for a different response of the examined hybrids to the conditions of drought or waterlogging may be a more economical water balance and more favourable relations between the shoot and root dimensions in the drought resistant genotype. The observed modifications of the internal root structure caused by water deficit in plant tissues may partly influence on water conductivity and transport within roots. The results suggest that the morphological and anatomical traits of the maize root system may be used in practice as direct or indirect selection criteria in maize breeding.     

Transcriptional regulation and functional properties of <Emphasis Type="Italic">Arabidopsis</Emphasis> Pht1;4, a high affinity transporter contributing greatly to phosphate uptake in phosphate deprived plants

Phosphate mobilization into the plant is a complex process requiring numerous transporters for absorption and translocation of this major nutrient. In the genome of Arabidopsis thaliana, nine closely related high affinity phosphate transporters have been identified but their specific roles remain unclear. Here we report the molecular, histological and physiological characterization of Arabidopsis pht1;4 high affinity phosphate transporter mutants. Using GUS-gene trap and in situ hybridization, Pht1;4 was found mainly expressed in inorganic phosphate (Pi) limiting medium in roots, primarily in the epidermis, the cortex and the root cap. In addition to this, expression was also observed at the lateral root branch points on the primary root and in the stele of lateral roots, suggesting a role of Pht1;4 in phosphate absorption and translocation from the growth medium to the different parts of the plant. Pi-starved pht1;4 plantlets exhibited a strong reduction of phosphate uptake capacity (40). This phenotype appears only related to the pht1;4 mutation as there were no obvious changes in the expression of other Pht1 family members in the mutants background. However, after 10 days of growth on phosphate deficient or sufficient medium, the Pi content in the mutants was not significantly different from that of the corresponding wild type controls. Furthermore, the mutants did not display any obvious growth defects or visible phenotypes when grown on a low phosphate containing medium. The work described here offers a first step in the complex genetic dissection of the phosphate transport system in planta.     

Heavy metals in white lupin: uptake, root-to-shoot transfer and redistribution within the plant

The translocation of manganese (Mn), nickel (Ni), cobalt (Co), zinc (Zn) and cadmium (Cd) in white lupin (Lupinus albus cv. Amiga) was compared considering root-to-shoot transport, and redistribution in the root system and in the shoot, as well as the content at different stages of cluster roots and in other roots. To investigate the redistribution of these heavy metals, lupin plants were labelled via the root for 24 h with radionuclides and subsequently grown hydroponically for several weeks. 54Mn, 63Ni and 65Zn were transported via the xylem to the shoot. 63Ni and 65Zn were redistributed afterwards via the phloem from older to younger leaves, while 54Mn remained in the oldest leaves. A strong retention in the root was observed for 57Co and 109Cd. Cluster roots contained higher concentrations of all heavy metals than noncluster roots. Concentrations were generally higher at the beginning of cluster root development (juvenile and immature stages). Mature cluster roots also contained high levels of 54Mn and 57Co, but only reduced concentrations of 63Ni, 65Zn and 109Cd.     

QTL analysis of cadmium and zinc accumulation in the heavy metal hyperaccumulator Thlaspi caerulescens

Thlaspi caerulescens (Tc; 2n = 14) is a natural Zn, Cd and Ni hyperaccumulator species belonging to the Brassicaceae family. It shares 88% DNA identity in the coding regions with Arabidopsis thaliana (At) (Rigola et al. 2006). Although the physiology of heavy metal (hyper)accumulation has been intensively studied, the molecular genetics are still largely unexplored. We address this topic by constructing a genetic map based on AFLP markers and expressed sequence tags (ESTs). To establish a genetic map, an F(2) population of 129 individuals was generated from a cross between a plant from a Pb/Cd/Zn-contaminated site near La Calamine, Belgium, and a plant from a comparable site near Ganges (GA), France. These two accessions show different degrees of Zn and, particularly, Cd accumulation. We analyzed 181 AFLP markers (of which 4 co-dominant) and 13 co-dominant EST sequences-based markers and mapped them to seven linkage groups (LGs), presumably corresponding to the seven chromosomes of T. caerulescens. The total length of the genetic map is 496 cM with an average density of one marker every 2.5 cM. This map was used for Quantitative Trait Locus (QTL) mapping in the F(2). For Zn as well as Cd concentration in root we mapped two QTLs. Three QTLs and one QTL were mapped for Zn and Cd concentration in shoot, respectively. These QTLs explain 23.8-60.4% of the total variance of the traits measured. We found only one common locus (LG6) for Zn and Cd (concentration in root) and one common locus for shoot and root concentrations of Zn (LG1) and of Cd (LG3). For all QTLs, the GA allele increased the trait value except for two QTLs for Zn accumulation in shoot (LG1 and LG4) and one for Zn concentration in root (LG1).