Search

Context Analytical procedures and technologies for soil analyses can be prohibitively expensive for small laboratories and researchers in developing countries. There is a need for low cost and high-throughput methods for assaying pH, extractable aluminium and phosphorus when conducting field trials on acid soils. Aims We investigated methods to develop rapid yet inexpensive colorimetric assays for the assay of pH, extractable aluminium and Colwell phosphorus in soil extracts. Methods We developed a colorimetric method to measure soil pH enabling pH to be quantified in a high-throughput assay. Similarly, two existing methods for extractable aluminium and Colwell P were modified for high throughput assays also using microtiter plates. Key results All three methods yielded linear relationships when using absorbance to quantify the parameters with the high throughput methods. Furthermore, there was a strong correlation between pH values of the soil samples obtained with the colorimetric assay and pH values measured with a glass electrode. Conclusions We demonstrated that the rapid assays for all three methods can be implemented to characterise field sites through the mapping of distributions for extractable Al, Colwell P and pH. Implications The high-throughput methods described here will be useful for researchers who conduct field trials to map variations in soil pH, soluble Al and Colwell P. Although the focus of the current work was on acid soils, the colorimetric pH and Colwell P methods can also be applied to non-acid soils.

Purpose Durum wheat is sensitive of acid soils because it lacks effective genes for Al3+ tolerance. Previous research showed introgression of the TaMATE1B and TaALMT1 genes individually increased the Al3+ tolerance of durum wheat. Here we aimed to (a) combine the genes into a single durum line, (b) compare the various introgression lines and (c) establish the effectiveness of the introgressions in improving the acid soil tolerance in the field. Methods Durum wheat lines homozygous for Al3+-tolerant alleles of TaMATE1B and TaALMT1 were crossed to develop a line that incorporated both genes. The parental cultivar, lines with the individual genes and the line with both genes introgressed were screened for Al3+ tolerance by hydroponic and soil cultures in a growth cabinet. The lines were also assessed for biomass production and grain yield in the field on acid soils. Results The durum wheat lines with the various Al3+-tolerance genes introgressed performed better based on root growth than Jandaroi, the parental cultivar, in both hydroponic and soil assays when grown in a cabinet. The various introgression lines were tolerant of acid soils compared to Jandaroi when grown in the field as assessed by shoot biomass and grain yield. Conclusion The TaALMT1 and TaMATE1B genes improve the acid soil tolerance of durum wheat with indications that combining both genes is the most effective strategy. The various lines will be valuable to breeders who wish to enhance the acid soil tolerance of durum germplasm.

Certain soil microorganisms can improve plant growth, and practices that encourage their proliferation around the roots can boost production and reduce reliance on agrochemicals. The beneficial effects of the microbial inoculants currently used in agriculture are inconsistent or short-lived because their persistence in soil and on roots is often poor. A complementary approach could use root exudates to recruit beneficial microbes directly from the soil and encourage inoculant proliferation. However, it is unclear whether the release of common organic metabolites can alter the root microbiome in a consistent manner and if so, how those changes vary throughout the whole root system. In this study, we altered the expression of transporters from the ALUMINUM-ACTIVATED MALATE TRANSPORTER and the MULTIDRUG AND TOXIC COMPOUND EXTRUSION families in rice (Oryza sativa L.) and wheat (Triticum aestivum L.) and tested how the subsequent release of their substrates (simple organic anions, including malate, citrate, and γ-amino butyric acid) from root apices affected the root microbiomes. We demonstrate that these exudate compounds, separately and in combination, significantly altered microbiome composition throughout the root system. However, the root type (seminal or nodal), position along the roots (apex or base), and soil type had a greater influence on microbiome structure than the exudates. These results reveal that the root microbiomes of important cereal species can be manipulated by altering the composition of root exudates, and support ongoing attempts to improve plant production by manipulating the root microbiome., One-sentence summary: The root microbiome of rice and wheat can be manipulated by altering the activity of root transporters and exudates.

High aluminum (Al3+) concentrations associated with subsoil acidity is a major constraint to durum wheat (Triticum turgidum) production as it inhibits root growth affecting crop tolerance to drought. This study evaluated the introgressed TaMATE1B gene on drought resistance and Al3+ toxicity in durum wheat. Durum wheat lines Jandaroi–TaMATE1B (introgressed with the TaMATE1B gene) and Jandaroi–null (without TaMATE1B gene) were grown in 1-m deep columns filled with re-constructed field soil with Al3+-rich acid subsoil in a glasshouse under well-watered conditions until the onset of ear emergence (Z51), before imposing well-watered and terminal drought treatments. Jandaroi–TaMATE1B produced 25.3 % higher grain yield than Jandaroi–null under well-watered conditions and 49.0 % higher grain yield under terminal drought. Terminal drought reduced grain yield by 47.7 % in Jandaroi–TaMATE1B and 72 % in Jandaroi–null, relative to well-watered conditions. The effects of TaMATE1B on grain yield can be attributed to increased root growth and proliferation below 0.4 m in Al3+-toxic soil. Jandaroi–TaMATE1B had 34.5 and 32.0 % more total root biomass than Jandaroi–null in the well-watered and terminal drought treatments, respectively (P ≤ 0.05). Jandaroi–TaMATE1B had a significantly higher root: shoot ratio than Jandaroi–null at Z51. Introgression of the TaMATE1B gene did not affect grain-filling duration, but terminal drought reduced it by 24 days in both lines. Introgression of the Al3+-tolerant TaMATE1B gene into durum wheat improved terminal drought resistance by enabling root growth and proliferation into deep layers of Al3+-rich acidic soil.

Understanding how climate change affects the phosphorus (P) nutrition of crops grown on acid soils is important in optimizing the management of P, and to secure future food production on these soils. This study assessed the impact of elevated COElevated COElevated CO

Purpose Organic anions commonly released from plant roots are widely reported to mobilize soil phosphorus (P). We characterized soil organic P that was mobilized by organic anions and assessed its amenability to hydrolysis by phosphatase enzymes. Methods Six soils differing in organic P content were extracted with citrate, malate or oxalate solutions and incubated with preparations of phosphomonoesterase, phosphodiesterase, or phytase. Organic P compounds present in these extracts were putatively identified and quantified with solution 31P NMR spectroscopy and the enzyme-labile P fractions were assessed by changes in molybdate reactive P (MRP) concentration. Results Organic P mobilization varied markedly among the organic anions. Extraction with 10 mM citrate was most effective and extracted 7.8-fold more total P than the water controls across all soils. Approximately 95% of the extracted P was non-MRP. The organic anions increased both the amount of P extracted and the proportion of the total extracted P that was phosphatase labile. Phytase was generally the most effective enzyme with up to 60% of the total non-MRP being amenable to hydrolysis by phytase across all extracts. The presence of inositol hexakisphosphates in the extracts, as well as other forms of organic P including nucleic acids and phospholipids, was verified by 31P NMR with concentrations dependent on both organic anion and soil type. Conclusion The combination of organic anions and phosphatases represents a key mechanism by which plants and microorganisms can enhance the bioavailability of soil P. This has important implications for understanding P dynamics in natural and managed ecosystems and for ongoing efforts to improve the P-use efficiency of agricultural plants.

Six cycles of recurrent selection for early shoot vigour in wheat resulted in significant increases in leaf width and shoot biomass. Here, in replicated controlled-environment studies, the effect of early shoot vigour on root biomass, rhizosheath size, root hair length, and cell size in the roots and leaves was examined across different cycles of selection. Increased shoot vigour was associated with greater root biomass, larger rhizosheath size, and longer root hairs. Our findings demonstrate that rhizosheath size was a reliable surrogate for root hair length in this germplasm. Examination of the root epidermis revealed that the ‘cell body’ of the trichoblasts (hair-forming cells) and the atrichoblasts (non-hair-forming cells) decreased in size as shoot vigour increased. Therefore, in higher vigour germplasm, longer root hairs emerged from smaller trichoblasts so that total trichoblast volume (root hair plus cell body) was generally similar regardless of shoot vigour. Similarly, the sizes of the four main cell types on the leaf epidermis became progressively smaller as shoot vigour increased, which also increased stomatal density. The relationship between shoot vigour and root traits is considered, and the potential contribution of below-ground root traits to performance and competitiveness of high vigour germplasm is discussed.

Subsoil acidity with a high aluminium (Al3+) soil content inhibits root growth and proliferation of durum wheat (tetraploid AABB, Triticum turgidum) leading to poor nutrient and water uptake. This study evaluated the impact of Al3+-tolerantTaMATE1B allele on root and shoot traits of durum wheat grown in an acidic soil with a high Al3+concentration. Two durum wheat lines, Jandaroi–TaMATE1B with the TaMATE1B gene introgressed from Al3+-tolerant bread wheat and Jandaroi–null (a sister line lacking the Al3+-tolerant TaMATE1B allele), were grown in rhizoboxes in a glasshouse. We mapped root growth and proliferation over time and measured shoot traits and grain yield. Introgression of the Al3+-tolerant TaMATE1B allele into durum wheat enabled root growth and proliferation below 0.25 m of the soil profile, where the soil pH was low (4.1, CaCl2 extract) with high Al3+ content (16.5 mg kg−1), and increased total root length and biomass at 42 days after sowing (DAS; Z33) by 38.3 and 22%, respectively, relative to the Jandaroi–null. Differences in root growth between the two lines were apparent from tillering stage (Z33) and by 50% anthesis (Z64), respectively. Jandaroi–TaMATE1B had 69.2% greater root biomass, 76.2% greater root length, 5.89% greater leaf area and 18% greater shoot biomass than Jandaroi–null at 50% anthesis (Z64). Time to anthesis and physiological maturity was delayed 6–7 days in Jandaroi–TaMATE1B, compared to Jandaroi–null. Jandaroi–TaMATE1B tended to have relatively greater, but not significantly different, shoot biomass, grain yield and yield components than Jandaroi–null. Introgression of the Al3+-tolerant TaMATE1B allele into durum wheat enabled root growth and proliferation down an acidic soil profile with a high Al3+ concentration. We assume that in the field where plants need to acquire water at depth differences in above-ground parameters would be amplified.

© 2018, Springer International Publishing AG, part of Springer Nature. Background & aim: It is unclear how elevated CO2 (eCO2) affects the response of crops to soil acidity. This study examined the effect of eCO2 on acid-soil tolerance of hexaploid wheat genotypes that vary in Al3+ resistance due to differences in root efflux of citrate and malate. Methods: Three pairs of near-isogenic lines were grown for 24–25 days under ambient CO2 (400 µmol mol-1) and eCO2 (800 µmol mol-1) in acid soils and hydroponics with various Al3+ concentrations. The lines consisted of pairs that differed in alleles of the TaALMT1 and TaMATE1B genes. Plant growth parameters and rhizosphere soil properties were measured. Results: Elevated CO2 increased the slope of negative correlations between root and shoot biomass and Al3+ concentration in the rhizosphere of a line that has an Al3+-sensitive allele of the TaALMT1 gene conditioning malate efflux (ES8), but did not change that of a near-isogenic sister line with an Al3+-resistant TaALMT1 allele (ET8). Elevated CO2 decreased the relative root length and biomass (% of limed soil) of a line that lacked both malate and citrate efflux (Egret), but did not affect lines that possessed either malate or citrate efflux. Elevated CO2 had no effect on either malate or citrate efflux from root tips of Al3+-resistant lines. Elevated CO2 also increased Al3+ concentration and decreased NH4+ concentration in rhizosphere soil, but decreased concentrations of Al and Zn in shoots. Conclusions: Elevated CO2 decreased acid-soil tolerance of Al3+sensitive genotypes but not of Al3+ resistant genotypes. Malate efflux played a dominant role in conferring acid-soil tolerance to hexaploid wheat.

Phosphorus (P) is an essential plant nutrient and regular applications are essential in most farming systems to maintain high yields. Yet the P fertilizers applied to crops and pastures are derived from non-renewable resources. It is therefore important to find agronomic and genetic strategies for using this resource efficiently, especially since only a proportion of the applied P is absorbed by crops. The aim of this study was to identify Quantitative Trait Loci (QTL) for P use efficiency (PUE) in wheat using a Multiparent Advanced Generation InterCross (MAGIC) population grown in the field. The 357 genotypes were arranged in paired plots with and without P fertilization. Yield and biomass were measured and PUE was calculated as either the performance of the genotype relative to the average response to fertilization, or the performance of the genotype relative to the average resilience in the absence of fertilization. Five trials were conducted over three years in Australia at three sites with contrasting clay and sandy soil types.Genotypic variation for response and resilience were identified in all trials with moderate to strong correlation with productivity with and without P between trials. Multiparent Whole Genome Average Interval Mapping (WGAIM) QTL analyses were conducted on the four traits (Biomass / Yield × P Response / Resilience) across the five trials and identified 130 QTL in total. QTL within 10 cM of each other were clustered into 56 groups that were likely to represent identical or linked loci. Of the clusters 27 (48%) contained only a single QTL but 17 (31%) contained 3 or more in different trials or traits. This suggests multiple biological mechanisms driving PUE in different environments. Eight of the 56 groups collocated with QTL for seedling root hair length identified in the same MAGIC population in an earlier study.HighlightIdentification of genetic loci for phosphorus use efficiency in a multigenic population of Australian wheats grown on contrasting soils.

Elevated atmospheric CO

Mutants without root hairs show reduced inorganic orthophosphate (Pi) uptake and compromised growth on soils when Pi availability is restricted. What is less clear is whether root hairs that are longer than wild-type provide an additional benefit to phosphorus (P) nutrition. This was tested using transgenic Brachypodium lines with longer root hairs. The lines were transformed with the endogenous BdRSL2 and BdRSL3 genes using either a constitutive promoter or a root hair-specific promoter. Plants were grown for 32 d in soil amended with various Pi concentrations. Plant biomass and P uptake were measured and genotypes were compared on the basis of critical Pi values and P uptake per unit root length. Ectopic expression of RSL2 and RSL3 increased root hair length three-fold but decreased plant biomass. Constitutive expression of BdRSL2, but not expression of BdRSL3, consistently improved P nutrition as measured by lowering the critical Pi values and increasing Pi uptake per unit root length. Increasing root hair length through breeding or biotechnology can improve P uptake efficiency if the pleotropic effects on plant biomass are avoided. Long root hairs, alone, appear to be insufficient to improve Pi uptake and need to be combined with other traits to benefit P nutrition.

Made available in DSpace on 2018-12-11T17:35:24Z (GMT). No. of bitstreams: 0 Previous issue date: 2018-03-01 Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Aim: A major limitation to plant growth in acid soils is the prevalence of toxic Al3+. Most genotypic variation for acid soil-tolerance in wheat is linked with the Al3+-activated efflux of malate anions from roots which is controlled by TaALMT1 on chromosome 4DL. Recent studies have also linked TaALMT1 with tolerance to high pH solutions and alkaline soils. This study tested the hypothesis that an Al3+-resistant allele of TaALMT1 also confers tolerance to alkaline conditions. Methods: The near-isogenic wheat lines, ET8 (Al3+-resistant) and ES8 (Al3+-sensitive), have different alleles of the TaALMT1 gene and contrasting resistance to Al3+ toxicity. Growth of these lines was compared in acid and alkaline soils with contrasting mineralogy and in a range of high pH hydroponic solutions of varying composition. Results: No consistent differences in root or shoot growth were detected between the lines in the alkaline soils or in the high pH hydroponic treatments. Malate efflux was detected from ET8 in acidic solution with Al3+ but no substantial malate efflux was detected at pH 9.0 treatment with added Na2SO4. Conclusion: The results are inconsistent with the hypothesis that the TaALMT1 gene confers an advantage to wheat on alkaline soils. CSIRO Agriculture and Food, GPO Box 1700 Botanics department Sao Paulo State University UNESP Chengdu Institute of Biology Chinese Academy of Sciences Botanics department Sao Paulo State University UNESP

Aluminum-activated malate transporters (ALMTs) form a family of anion channels in plants, but little is known about most of its members. This study examined the function of OsALMT4 from rice (Oryza sativa). We show that OsALMT4 is expressed in roots and shoots and that the OsALMT4 protein localizes to the plasma membrane. Transgenic rice lines overexpressing (OX) OsALMT4 released malate from the roots constitutively and had 2-fold higher malate concentrations in the xylem sap than nulls, indicating greater concentrations of malate in the apoplast. OX lines developed brown necrotic spots on the leaves that did not appear on nulls. These symptoms were not associated with altered concentrations of any mineral element in the leaves, although the OX lines had higher concentrations of Mn and B in their grain compared with nulls. While total leaf Mn concentrations were not different between the OX and null lines, Mn concentrations in the apoplast were greater in the OX plants. The OX lines also displayed increased expression of Mn transporters and were more sensitive to Mn toxicity than null plants. We showed that the growth of wild-type rice was unaffected by 100 µm Mn in hydroponics but, when combined with 1 mm malate, this concentration inhibited growth. We conclude that increasing OsALMT4 expression affected malate efflux and compartmentation within the tissues, which increased Mn concentrations in the apoplast of leaves and induced the toxicity symptoms. This study reveals new links between malate transport and mineral nutrition.

Background The structure and function of plant roots and their interactions with soil are exciting scientific frontiers that will ultimately reveal much about our natural systems, global water and mineral and carbon cycles, and help secure food supplies into the future. This Special Issue presents a collection of papers that address topics at the forefront of our understanding of root biology. Scope These papers investigate how roots cope with drought, nutrient deficiencies, toxicities and soil compaction as well as the interactions that roots have with soil microorganisms. Roots of model plant species, annual crops and perennial species are studied in short-term experiments through to multi-year trials. Spatial scales range from the gene up to farming systems and nutrient cycling. The diverse, integrated approaches described by these studies encompass root genetics as applied to soil management, as well as documenting the signalling processes occurring between roots and shoots and between roots and soil. Conclusions This Special Issue on roots presents invited reviews and research papers covering a span of topics ranging from fundamental aspects of anatomy, growth and water uptake to roots in crop and pasture systems. Understanding root structure and function and adaptation to the abiotic and biotic stresses encountered in field conditions is important for sustainable agricultural production and better management of natural systems.

Elevated atmospheric CO2 (eCO2) concentration can increase root exudation into soils, which improves plant tolerance to abiotic stresses. This review used a meta-analysis to assess effect sizes of eCO2 on both efflux rates and total amounts of some specific root exudates, and dissected whether eCO2 enhances plant's resistance to nutrient deficiency and ion toxicity via root exudates. Elevated CO2 did not affect efflux rates of total dissolved organic carbon, a measure of combined root exudates per unit of root biomass or length, but increased the efflux amount of root systems per plant by 31% which is likely attributed to increased root biomass (29%). Elevated CO2 increased efflux rates of soluble-sugars, carboxylates, and citrate by 47%, 111%, and 16%, respectively, but did not affect those of amino acids and malate. The increased carbon allocation to roots, increased plant requirements of mineral nutrients, and heightened detoxification responses to toxic ions under eCO2 collectively contribute to the increased efflux rates despite lacking molecular evidence. The increased efflux rates of root exudates under eCO2 were closely associated with improved nutrient uptake whilst less studies have validated the associations between root exudates and resistance to toxic ions of plants when grown under eCO2. Future studies are required to reveal how climate change (eCO2) affect the efflux of specific root exudates, particularly organic anions, the corresponding nutrient uptake and toxic ion resistance from plant molecular biology and soil microbial ecology perspectives.

BACKGROUND AND AIMS: Soil acidity currently limits root growth and crop production in many regions, and climate change is leading to uncertainties regarding future food supply. However, it is unknown how elevated CO(2) (eCO(2)) affects the performance of wheat crops in acid soils under field conditions. We investigated the effects of eCO(2) on plant growth and yield of three pairs of near-isogenic hexaploid wheat lines differing in alleles of aluminium-resistant genes TaALMT1 (conferring root malate efflux) and TaMATE1B (conferring citrate efflux). METHODS: Plants were grown until maturity in an acid soil under ambient CO(2) (aCO(2); 400 µmol mol(−1)) and eCO(2) (550 µmol mol(−1)) in a soil free-air CO(2) enrichment facility (SoilFACE). Growth parameters and grain yields were measured. KEY RESULTS: Elevated CO(2) increased grain yield of lines carrying TaMATE1B by 22 % and lines carrying only TaALMT1 by 31 %, but did not increase the grain yield of Al(3+)-sensitive lines. Although eCO(2) promoted tiller formation, coarse root length and root biomass of lines carrying TaMATE1B, it did not affect ear number, and it therefore limited yield potential. By contrast, eCO(2) decreased or did not change these parameters for lines carrying only TaALMT1, and enhanced biomass allocation to grains thereby resulting in increased grain yield. Despite TaMATE1B being less effective than TaALMT1 at conferring Al(3+) resistance based on root growth, the gene promoted grain yield to a similar level to TaALMT1 when the plants were grown in acid soil. Furthermore, TaALMT1 and TaMATE1B were not additive in their effects. CONCLUSIONS: As atmospheric CO(2) increases, it is critical that both Al(3+)-resistance genes (particularly TaALMT1) should be maintained in hexaploid wheat germplasm in order for yield increases from CO(2) fertilization to be realized in acid soils.

Genetic modification of shoot and root morphology has potential to improve water and nutrient\ud 19 uptake of wheat crops in rainfed environments. Near-isogenic lines (NILs) varying for a tillering\ud 20 inhibition (tin) gene and representing multiple genetic backgrounds were investigated in contrasting\ud 21 controlled environments for shoot and root growth. Leaf area, shoot and root biomass were similar\ud 22 until tillering whereupon reduced tillering in tin-containing NILs produced reductions of up to 60% in\ud 23 total leaf area and biomass, and increases in total root length of up to 120% and root biomass to\ud 24 145%. Together, root-to-shoot ratio increased two-fold with the tin gene. The influence of tin on shoot\ud 25 and root growth was greatest in the cv. Banks genetic background, particularly in the biculm-selected\ud 26 NIL, and was typically strongest in cooler environments. A separate de-tillering study confirmed\ud 27 greater root-to-shoot ratios with regular tiller removal in non-tin containing genotypes. In validating\ud 28 these observations in a rainfed field study, the tin allele had a negligible effect on seedling growth but\ud 29 was associated with significantly (P

Highlight: Early vigour in wheat was shown to be a valuable trait for improving biomass accumulation and phosphate uptake from a low-P soil., Quantitative trait loci (QTLs) for shoot biomass were identified in wheat grown on a soil high in total phosphorus (P) but low in plant-available P. The two populations screened included recombinant inbred lines (RILs) from Chuan-Mai 18/Vigour 18 and doubled-haploid lines from Kukri/Janz. Glasshouse-grown plants were harvested at the five-leaf stage. Seven QTLs for shoot biomass were identified in the RILs, with the largest on chromosome 7A accounting for 7.4% of the phenotypic variance. RILs from the upper tail had larger embryos than RILs from the lower tail. Tail lines were then grown in non-limiting P and the results indicated that early vigour and the capacity to access P contributed to the initial distribution. The influence of early vigour on P nutrition was examined further with advanced vigour lines (AVLs). The AVLs accumulated more shoot biomass, maintained lower shoot P concentrations, and showed greater P-acquisition efficiency than Vigour 18. Nine QTLs for shoot biomass were identified in the Kukri/Janz population. Two on chromosomes 4B and 4D accounted for 24.8% of the variance. Candidates underlying these QTLs are the Rht genes. We confirmed the influence of these genes using near-isogenic lines with different Rht alleles. The dwarf and semi-dwarf alleles affected shoot and root biomass at high and low P but not the efficiency of P acquisition. We conclude that early vigour contributed to the distributions in both populations. Early vigour can increase plant growth at suboptimal P and some sources can also improve the efficiency of P acquisition.

Highlight Rhizosheath size of wheat was used as a surrogate for root hair length to map QTL in lines derived from multiparent advanced generation intercrosses., Rhizosheaths comprise soil that adheres to plant roots and, in some species, are indicative of root hair length. In this study, the genetics of rhizosheath size in wheat was investigated by screening the progeny of multiparent advanced generation intercrosses (MAGIC). Two MAGIC populations were screened for rhizosheath size using a high throughput method. One MAGIC population was developed from intercrosses between four parents (4-way) and the other from intercrosses between eight parents (8-way). Transgressive segregation for rhizosheath size was observed in both the 4-way and 8-way MAGIC populations. A quantitative trait loci (QTL) analysis of the 4-way population identified six major loci located on chromosomes 2B, 4D, 5A, 5B, 6A, and 7A together accounting for 42% of the variation in rhizosheath size. Rhizosheath size was strongly correlated with root hair length and was robust across different soil types in the absence of chemical constraints. Rhizosheath size in the MAGIC populations was a reliable surrogate for root hair length and, therefore, the QTL identified probably control root hair elongation. Members of the basic helix-loop-helix family of transcription factors have previously been identified to regulate root hair length in Arabidopsis and rice. Since several wheat members of the basic helix-loop-helix family of genes are located within or near the QTL, these genes are candidates for controlling the long root hair trait. The QTL for rhizosheath size identified in this study provides the opportunity to implement marker-assisted selection to increase root hair length for improved phosphate acquisition in wheat.

The original version of this article unfortunately contained a mistake. The online supplementary material (composed of Table S1, Fig. S1, and Fig. S2) was omitted in the published online version. The supplementary material is available in this version.

Background and Aims: Expected increases in world population will continue to make demands on agricultural productivity and food supply. These challenges will only be met by increasing the land under cultivation and by improving the yields obtained on existing farms. Genetic engineering can target key traits to improve crop yields and to increase production on marginal soils. Soil acidity is a major abiotic stress that limits plant production worldwide. The goal of this study was to enhance the acid soil tolerance of wheat by increasing its resistance to Al3+ toxicity. Methods: Particle bombardment was used to transform wheat with TaALMT1, the Al3+ resistance gene from wheat, using the maize ubiquitin promoter to drive expression. TaALMT1 expression, malate efflux and Al3+ resistance were measured in the T1 and T2 lines and compared with the parental line and an Al3+-resistant reference genotype, ET8. Key Results: Nine T2 lines showed increased TaALMT1 expression, malate efflux and Al3+ resistance when compared with untransformed controls and null segregant lines. Some T2 lines displayed greater Al3+ resistance than ET8 in both hydroponic and soil experiments. Conclusions: The Al3+ resistance of wheat was increased by enhancing TaALMT1 expression with biotechnology. This is the first report of a major food crop being stably transformed for greater Al3+ resistance. Transgenic strategies provide options for increasing food supply on acid soils.

The aluminum (Al3+) tolerance of barley cultivars predominately from Brazil was compared to that of cultivars from other countries, wild barley accessions, and a transgenic line (L5) over-expressing TaALMT1, the major Al3+ tolerance gene from wheat. After screening conventional germplasm for Al3+ tolerance in hydroponics, 18 genotypes were further characterized in a short-term soil experiment. Among the Brazilian cultivars, Antarctica 01 and BRS Mariana showed the greatest relative root length (RRL) in acid soil. However, these cultivars were significantly less tolerant than the foreign cultivars Dayton (USA) and Murasakimochi (Japan) and the transgenic line L5 which out-performed all conventional genotypes. In long-term growth trials, the transgenic line produced the greatest relative root and relative shoot dry weight. Relative grain yield was greatest in the transgenic line and Dayton. All genotypes were also scored for two genetic markers linked to HvAACT1, the major Al3+ tolerance gene in barley. One marker detects a 1-kb insertion in the promoter that increases gene expression and leads to increased Al3+-activated citrate efflux from root apices. The other marker detects a 21-bp indel downstream of the coding region. The 1-kb insertion was only detected in Dayton and Murasakimochi that were the best performing cultivars among the non-transgenic germplasm. Interestingly, the Brazilian cultivars with an intermediate level of tolerance, Antarctica 01 and BRS Mariana, lacked the 1-kb insertion but had enhanced HvAACT1 expression compared to an Al3+-sensitive cultivar. No clear correlation was observed between Al3+ tolerance and the 21-bp indel marker in the short-term soil trials. We conclude that improved Al3+ tolerance in barley could be achieved by combining the best allele of HvAACT1 along with TaALMT1 as a transgene.

We examined the function of OsALMT4 in rice (Oryza sativa L.) which is a member of the aluminum-activated malate transporter family. Previous studies showed that OsALMT4 localizes to the plasma membrane and that expression in transgenic rice lines results in a constitutive release of malate from the roots. Here, we show that OsALMT4 is expressed widely in roots, shoots, flowers, and grain but not guard cells. Expression was also affected by ionic and osmotic stress, light and to the hormones ABA, IAA, and salicylic acid. Malate efflux from the transgenic plants over-expressing OsALMT4 was inhibited by niflumate and salicylic acid. Growth of transgenic lines with either increased OsALMT4 expression or reduced expression was measured in different environments. Light intensity caused significant differences in growth between the transgenic lines and controls. When day-time light was reduced from 700 to 300 μmol m-2s-1 independent transgenic lines with either increased or decreased OsALMT4 expression accumulated less biomass compared to their null controls. This response was not associated with differences in photosynthetic capacity, stomatal conductance or sugar concentrations in tissues. We propose that by disrupting malate fluxes across the plasma membrane carbon partitioning and perhaps signaling are affected which compromises growth under low light. We conclude that OsALMT4 is expressed widely in rice and facilitates malate efflux from different cell types. Altering OsALMT4 expression compromises growth in low-light environments.

Do root hairs help roots take up water from the soil? Despite the well-documented role of root hairs in phosphate uptake, their role in water extraction is controversial. We grew barley (Hordeum vulgare cv Pallas) and its root-hairless mutant brb in a root pressure chamber, whereby the transpiration rate could be varied whilst monitoring the suction in the xylem. The method provides accurate measurements of the dynamic relationship between the transpiration rate and xylem suction. The relationship between the transpiration rate and xylem suction was linear in wet soils and did not differ between genotypes. When the soil dried, the xylem suction increased rapidly and non-linearly at high transpiration rates. This response was much greater with the brb mutant, implying a reduced capacity to take up water. We conclude that root hairs facilitate the uptake of water by substantially reducing the drop in matric potential at the interface between root and soil in rapidly transpiring plants. The experiments also reinforce earlier observations that there is a marked hysteresis in the suction in the xylem when the transpiration rate is rising compared with when it is falling, and possible reasons for this behavior are discussed.

Background and Aims Long root hairs enable the efficient uptake of poorly mobile nutrients such as phosphorus. Mapping the chromosomal locations of genes that control root hair length can help exploit the natural variation within crops to develop improved cultivars. Genetic stocks of the wheat cultivar 'Chinese Spring' were used to map genes that control root hair length. Methods Aneuploid stocks of 'Chinese Spring' were screened using a rapid method based on rhizosheath size and then selected lines were assayed for root hair length to identify chromosomes harbouring genes controlling root hair length. A series of lines with various fractional deletions of candidate chromosomes were then screened to map the root hair loci more accurately. A line with a deletion in chromosome 5A was analysed with a 90 000 single nucleotide polymorphism (SNP) array. The phosphorus acquisition efficiency (PAE) of one deletion line was compared with that of euploid 'Chinese Spring' by growing the seedlings in pots at low and luxury phosphorus supplies. Key Results Chromosomes 1A, 1D and 5A were found to harbour genes controlling root hair length. The 90 000 SNP array identified two candidate genes controlling root hair length located on chromosome 5A. The line with a deletion in chromosome 5A had root hairs that were approx. 20 % shorter than euploid 'Chinese Spring', but this was insufficient to reduce its PAE. Conclusions A rapid screen for rhizosheath size enabled chromosomal regions controlling root hair length to be mapped in the wheat cultivar 'Chinese Spring' and subsequent analysis with an SNP array identified candidate genes controlling root hair length. The difference in root hair length between euploid 'Chinese Spring' and a deletion line identified in the rapid screen was still apparent, albeit attenuated, when the seedlings were grown on a fully fertilized soil.

Phosphorus (P) deficiency in some plant species triggers the release of organic anions such as citrate and malate from roots. These anions are widely suggested to enhance the availability of phosphate for plant uptake by mobilizing sparingly-soluble forms in the soil. Carazinho is an old wheat (Triticum aestivum) cultivar from Brazil, which secretes citrate constitutively from its root apices, and here we show that it also produces relatively more biomass on soils with low P availability than two recent Australian cultivars that lack citrate efflux. To test whether citrate efflux explains this phenotype, we generated two sets of near-isogenic lines that differ in citrate efflux and compared their biomass production in different soil types and with different P treatments in glasshouse experiments and field trials. Citrate efflux improved relative biomass production in two of six glasshouse trials but only at the lowest P treatments where growth was most severely limited by P availability. Furthermore, citrate efflux provided no consistent advantage for biomass production or yield in multiple field trials. Theoretical modeling indicates that the effectiveness of citrate efflux in mobilizing soil P is greater as the volume of soil into which it diffuses increases. As efflux from these wheat plants is restricted to the root apices, the potential for citrate to mobilize sufficient P to increase shoot biomass may be limited. We conclude that Carazinho has other attributes that contribute to its comparatively good performance in low-P soils.

Background and Aims Aluminium is toxic in acid soils because the soluble Al 3+ inhibits root growth. A mechanism of Al 3+ tolerance discovered in many plant species involves the release of organic anions from root apices. The Al 3+ -activated release of citrate from the root apices of Al 3+ -tolerant genotypes of barley is controlled by a MATE gene named HvAACT1 that encodes a citrate transport protein located on the plasma membrane. The aim of this study was to investigate whether expressing HvAACT1 with a constitutive promoter in barley and wheat can increase citrate efflux and Al 3+ tolerance of these important cereal species. Methods: HvAACT1 was over-expressed in wheat ( Triticum aestivum ) and barley ( Hordeum vulgare ) using the maize ubiquitin promoter. Root apices of transgenic and control lines were analysed for HvAACT1 expression and organic acid efflux. The Al 3+ tolerance of transgenic and control lines was assessed in both hydroponic solution and acid soil. Key Results and Conclusions: Increased HvAACT1 expression in both cereal species was associated with increased citrate efflux from root apices and enhanced Al 3+ tolerance, thus demonstrating that biotechnology can complement traditional breeding practices to increase the Al 3+ tolerance of important crop plants.

With the global population predicted to grow by at least 25 per cent by 2050, the need for sustainable production of nutritious foods is critical for human and environmental health. Recent advances show that specialized plant membrane transporters can be used to enhance yields of staple crops, increase nutrient content and increase resistance to key stresses, including salinity, pathogens and aluminium toxicity, which in turn could expand available arable land.

Phosphorus (P) is one of the essential nutrients for plants, and is indispensable for plant growth and development. P deficiency severely limits crop yield, and regular fertilizer applications are required to obtain high yields and to prevent soil degradation. To access P from the soil, plants have evolved high- and low-affinity Pi transporters and the ability to induce root architectural changes to forage P. Also, adjustments of numerous cellular processes are triggered by the P starvation response, a tightly regulated process in plants. With the increasing demand for food as a result of a growing population, the demand for P fertilizer is steadily increasing. Given the high costs of fertilizers and in light of the fact that phosphate rock, the source of P fertilizer, is a finite natural resource, there is a need to enhance P fertilizer use efficiency in agricultural systems and to develop plants with enhanced Pi uptake and internal P-use efficiency (PUE). In this review we will provide an overview of continuing relevant research and highlight different approaches towards developing crops with enhanced PUE. In this context, we will summarize our current understanding of root responses to low phosphorus conditions and will emphasize the importance of combining PUE with tolerance of other stresses, such as aluminum toxicity. Of the many genes associated with Pi deficiency, this review will focus on those that hold promise or are already at an advanced stage of testing (OsPSTOL1, AVP1, PHO1 and OsPHT1;6). Finally, an update is provided on the progress made exploring alternative technologies, such as phosphite fertilizer.

Summary •We found significant genetic variation in the ability of wheat (Triticum aestivum) to form rhizosheaths on acid soil and assessed whether differences in aluminium (Al3+) tolerance of root hairs between genotypes was the physiological basis for this genetic variation. •A method was developed to rapidly screen rhizosheath size in a range of wheat genotypes. Backcrossed populations were generated from cv Fronteira (large rhizosheath) using cv EGA-Burke (small rhizosheath) as the recurrent parent. •A positive correlation existed between rhizosheath size on acid soil and root hair length. In hydroponic experiments, root hairs of the backcrossed lines with large rhizosheaths were more tolerant of Al3+ toxicity than the backcrossed lines with small rhizosheaths. •We conclude that greater Al3+ tolerance of root hairs underlies the larger rhizosheath of wheat grown on acid soil. Tolerance of the root hairs to Al3+ was largely independent of the TaALMT1 gene which suggests that different genes encode the Al3+ tolerance of root hairs. The maintenance of longer root hairs in acid soils is important for the efficient uptake of water and nutrients.

Phosphorus (P)-deficiency is a significant challenge for agricultural productivity on many highly P-sorbing weathered and tropical soils throughout the world. On these soils it can be necessary to apply up to five-fold more P as fertiliser than is exported in products. Given the finite nature of global P resources, it is important that such inefficiencies be addressed. For low P-sorbing soils, P-efficient farming systems will also assist attempts to reduce pollution associated with P losses to the environment. P-balance inefficiency of farms is associated with loss of P in erosion, runoff or leaching, uneven dispersal of animal excreta, and accumulation of P as sparingly-available phosphate and organic P in the soil. In many cases it is possible to minimise P losses in runoff or erosion. Uneven dispersal of P in excreta typically amounts to ~5% of P-fertiliser inputs. However, the rate of P accumulation in moderate to highly P-sorbing soils is a major contributor to inefficient P-fertiliser use. We discuss the causal edaphic, plant and microbial factors in the context of soil P management, P cycling and productivity goals of farms. Management interventions that can alter P-use efficiency are explored, including better targeted P-fertiliser use, organic amendments, removing other constraints to yield, zone management, use of plants with low critical-P requirements, and modified farming systems. Higher productivity in low-P soils, or lower P inputs in fertilised agricultural systems can be achieved by various interventions, but it is also critically important to understand the agroecology of plant P nutrition within farming systems for improvements in P-use efficiency to be realised.

Salinity stress is a major limitation in barley production. Substantial genetic variation in tolerance occurs among genotypes of barley, so the development of salt-tolerant cultivars is a potentially effective approach for minimizing yield losses. The lack of economically viable methods for screening salinity tolerance in the field remains an obstacle to breeders, and molecular marker-assisted selection is a promising alternative. In this study, salinity tolerance of 172 doubled-haploid lines generated from YYXT (salinity-tolerant) and Franklin (salinity-sensitive) was assessed in glasshouse trials during the vegetative phase. A high-density genetic linkage map was constructed from 76 pairs of simple sequence repeats and 782 Diversity Arrays Technology markers which spanned a total of 1,147 cM. Five significant quantitative trait loci (QTL) for salinity tolerance were identified on chromosomes 1H, 2H, 5H, 6H and 7H, accounting for more than 50% of the phenotypic variation. The tolerant variety, YYXT, contributed the tolerance to four of these QTL and Franklin contributed the tolerance to one QTL on chromosome 1H. Some of these QTL mapped to genomic regions previously associated with salt tolerance in barley and other cereals. Markers associated with the major QTL identified in this study have potential application for marker-assisted selection in breeding for enhanced salt tolerance in barley.

Acid soils limit plant production worldwide because their high concentrations of soluble aluminium cations (Al(3+) ) inhibit root growth. Major food crops such as wheat (Triticum aestivum L.) have evolved mechanisms to resist Al(3+) toxicity, thus enabling wider distribution. The origins of Al(3+) resistance in wheat are perplexing because all progenitors of this hexaploid species are reportedly sensitive to Al(3+) stress. The large genotypic variation for Al(3+) resistance in wheat is largely controlled by expression of an anion channel, TaALMT1, which releases malate anions from the root apices. A current hypothesis proposes that the malate anions protect this sensitive growth zone by binding to Al(3+) in the apoplasm. We investigated the evolution of this trait in wheat, and demonstrated that it has multiple independent origins that enhance Al(3+) resistance by increasing TaALMT1 expression. One origin is likely to be Aegilops tauschii while other origins occurred more recently from a series of cis mutations that have generated tandemly repeated elements in the TaALMT1 promoter. We generated transgenic plants to directly compare these promoter alleles and demonstrate that the tandemly repeated elements act to enhance gene expression. This study provides an example from higher eukaryotes in which perfect tandem repeats are linked with transcriptional regulation and phenotypic change in the context of evolutionary adaptation to a major abiotic stress.

Acid soils restrict plant production around the world. One of the major limitations to plant growth on acid soils is the prevalence of soluble aluminium (Al(3+)) ions which can inhibit root growth at micromolar concentrations. Species that show a natural resistance to Al(3+) toxicity perform better on acid soils. Our understanding of the physiology of Al(3+) resistance in important crop plants has increased greatly over the past 20 years, largely due to the application of genetics and molecular biology. Fourteen genes from seven different species are known to contribute to Al(3+) tolerance and resistance and several additional candidates have been identified. Some of these genes account for genotypic variation within species and others do not. One mechanism of resistance which has now been identified in a range of species relies on the efflux of organic anions such as malate and citrate from roots. The genes controlling this trait are members of the ALMT and MATE families which encode membrane proteins that facilitate organic anion efflux across the plasma membrane. Identification of these and other resistance genes provides opportunities for enhancing the Al(3+) resistance of plants by marker-assisted breeding and through biotechnology. Most attempts to enhance Al(3+) resistance in plants with genetic engineering have targeted genes that are induced by Al(3+) stress or that are likely to increase organic anion efflux. In the latter case, studies have either enhanced organic anion synthesis or increased organic anion transport across the plasma membrane. Recent developments in this area are summarized and the structure-function of the TaALMT1 protein from wheat is discussed.

Members of the ALMT gene family contribute to the Al(3+) resistance of several plant species by facilitating malate efflux from root cells. The first member of this family to be cloned and characterized, TaALMT1, is responsible for most of the natural variation of Al(3+) resistance in wheat. The current study describes the isolation and characterization of HvALMT1, the barley gene with the greatest sequence similarity to TaALMT1. HvALMT1 is located on chromosome 2H which has not been associated with Al(3+) resistance in barley. The relatively low levels of HvALMT1 expression detected in root and shoot tissues were independent of external aluminium or phosphorus supply. Transgenic barley plants transformed with the HvALMT1 promoter fused to the green fluorescent protein (GFP) indicated that expression of HvALMT1 was relatively high in stomatal guard cells and in root tissues containing expanding cells. GFP fused to the C-terminus of the full HvALMT1 protein localized to the plasma membrane and motile vesicles within the cytoplasm. HvALMT1 conferred both inward and outward currents when expressed in Xenopus laevis oocytes that were bathed in a range of anions including malate. Both malate uptake and efflux were confirmed in oocyte assays using [(14)C]malate as a radiotracer. It is suggested that HvALMT1 functions as an anion channel to facilitate organic anion transport in stomatal function and expanding cells.

The effect of soil acidity on root and rhizosheath development in wheat and barley seedlings was investigated in an acid Ferrosol soil to which various amounts of lime (CaCO3) were applied to modify soil Al concentrations (pH (CaCl2): 4.22 to 5.35 and Al (CaCl2 extract): 17.7 to 0.4 mg kg−1 soil; respectively), and Ferrosol soil from an adjacent location at the same site which had a higher Al concentration (pH 4.19; 29.2 mg kg−1 Al). The cereal lines were selected on the basis of differences in their rate of root growth, Al-resistance and root hair morphology. Root morphology was assessed after 7 days of growth. The length of fine (mainly lateral) roots of Al-sensitive genotypes was more sensitive to soil Al concentrations than that of the coarse (mainly primary) roots. The experiments demonstrated that even where root growth was protected by expression of the TaALMT1 gene for Al-resistance, root-soil contact was diminished by soil acidity because root hair length (in many lines), and root hair density and rhizosheath formation (all lines) were adversely affected by soil acidity. In the case of Al-sensitive lines, fine root growth and rhizosheath mass were reduced over much the same range of soil Al concentrations (i.e. >3–6 mg kg−1 Al). Although Al-resistant lines could maintain fine root length under these conditions, they were similarly unable to maintain rhizosheath mass. This finding may help to explain why Al-resistant wheats which yield relatively well in deep acid soils, may also benefit from application of lime to the surface layers of the soil.

Barley (Hordeum vulgare L.), genetically modified with the Al(3+) resistance gene of wheat (TaALMT1), was compared with a non-transformed sibling line when grown on an acidic and highly phosphate-fixing ferrosol supplied with a range of phosphorus concentrations. In short-term pot trials (26 days), transgenic barley expressing TaALMT1 (GP-ALMT1) was more efficient than a non-transformed sibling line (GP) at taking up phosphorus on acid soil, but the genotypes did not differ when the soil was limed. Differences in phosphorus uptake efficiency on acid soil could be attributed not only to the differential effects of aluminium toxicity on root growth between the genotypes, but also to differences in phosphorus uptake per unit root length. Although GP-ALMT1 out-performed GP on acid soil, it was still not as efficient at taking up phosphorus as plants grown on limed soil. GP-ALMT1 plants grown in acid soil possessed substantially smaller rhizosheaths than those grown in limed soil, suggesting that root hairs were shorter. This is a probable reason for the lower phosphorus uptake efficiency. When grown to maturity in large pots, GP-ALMT1 plants produced more than twice the grain as GP plants grown on acid soil and 80% of the grain produced by limed controls. Expression of TaALMT1 in barley was not associated with a penalty in either total shoot or grain production in the absence of Al(3+), with both genotypes showing equivalent yields in limed soil. These findings demonstrate that an important crop species can be genetically engineered to successfully increase grain production on an acid soil.

The first confirmed mechanism for aluminum (Al) resistance in plants is encoded by the wheat (Triticum aestivum) gene, TaALMT1, on chromosome 4DL. TaALMT1 controls the Al-activated efflux of malate from roots, and this mechanism is widespread among Al-resistant genotypes of diverse genetic origins. This study describes a second mechanism for Al resistance in wheat that relies on citrate efflux. Citrate efflux occurred constitutively from the roots of Brazilian cultivars Carazinho, Maringa, Toropi, and Trintecinco. Examination of two populations segregating for this trait showed that citrate efflux was controlled by a single locus. Whole-genome linkage mapping using an F2 population derived from a cross between Carazinho (citrate efflux) and the cultivar EGA-Burke (no citrate efflux) identified a major locus on chromosome 4BL, Xcec, which accounts for more than 50% of the phenotypic variation in citrate efflux. Mendelizing the quantitative variation in citrate efflux into qualitative data, the Xcec locus was mapped within 6.3 cM of the microsatellite marker Xgwm495 locus. This linkage was validated in a second population of F2:3 families derived from a cross between Carazinho and the cultivar Egret (no citrate efflux). We show that expression of an expressed sequence tag, belonging to the multidrug and toxin efflux (MATE) gene family, correlates with the citrate efflux phenotype. This study provides genetic and physiological evidence that citrate efflux is a second mechanism for Al resistance in wheat.

Highlight Large rhizosheaths on wheat genotypes grown on acid soils improved the phosphorus acquisition efficiency compared with genotypes with small rhizosheaths. The rhizosheath trait was mapped to five major quantitative trait loci with largely additive genetic effect., Rhizosheaths comprise soil bound to roots, and in wheat (Triticum aestivum L.) rhizosheath size correlates with root hair length. The aims of this study were to determine the effect that a large rhizosheath has on the phosphorus (P) acquisition by wheat and to investigate the genetic control of rhizosheath size in wheat grown on acid soil. Near-isogenic wheat lines differing in rhizosheath size were evaluated on two acid soils. The soils were fertilized with mineral nutrients and included treatments with either low or high P. The same soils were treated with CaCO3 to raise the pH and detoxify Al3+. Genotypic differences in rhizosheath size were apparent only when soil pH was low and Al3+ was present. On acid soils, a large rhizosheath increased shoot biomass compared with a small rhizosheath regardless of P supply. At low P supply, increased shoot biomass could be attributed to a greater uptake of soil P, but at high P supply the increased biomass was due to some other factor. Generation means analysis indicated that rhizosheath size on acid soil was controlled by multiple, additive loci. Subsequently, a quantitative trait loci (QTL) analysis of an F6 population of recombinant inbred lines identified five major loci contributing to the phenotype together accounting for over 60% of the total genetic variance. One locus on chromosome 1D accounted for 34% of the genotypic variation. Genetic control of rhizosheath size appears to be relatively simple and markers based on the QTL provide valuable tools for marker assisted breeding.

The wheat ALMT1 gene encodes an aluminum (Al)-activated malate transport protein which confers Al-resistance. We investigated the membrane topology of this plasma-membrane localized protein with immunocytochemical techniques. Several green fluorescent protein (GFP)-fused and histidine (His)-tagged chimeras of ALMT1 were prepared based on a computer-predicted secondary structure and transiently expressed in cultured mammalian cells. Antibodies raised to polypeptide epitopes of ALMT1 were used in conjunction with the antibody to the His-tags to determine the topology of ALMT1. This study shows that the ALMT1 protein contains six transmembrane domains with the amino and carboxyl termini located on the extracellular side of the plasma membrane.