Hayato Maruyama

Overexpression of the LASAP2 gene for secretory acid phosphatase in white lupin improves the phosphorus uptake and growth of tobacco plants

Abstract Secretion of acid phosphatase (APase) from the roots to take up phosphorus (P) is a well-known strategy of plants under P-deficient conditions. White lupin, which shows vigorous growth in low-P soils, is noted for its ability to secrete APase under P-deficient conditions. The APase secreted by white lupin roots is stable in soil solution and shows low substrate specificity, suggesting that genetic modification of plants using the APase gene LASAP2 might improve their ability to use organic P. The objective of the present study was to evaluate the potential of LASAP2 transgenic plants to increase organic P utilization. Dry matter production and P accumulation were higher in LASAP2 transgenic tobacco plants grown in gel media containing soluble phytate as the sole P source than in wild-type tobacco plants. Phosphorus uptake by the transgenic plants also increased in soil culture conditions. LASAP2 was apparently more effective in the liberation of organic P, including phytate, in the soil than the native tobacco APase. Thus, the enzymatic stability of LASAP2 in the soil appears to be an important factor for P acquisition.

Localization of acid phosphatase activities in the roots of white lupin plants grown under phosphorus-deficient conditions

Abstract Acid phosphatase (APase) produced by the cluster roots of white lupin (Lupinus albus L.) plays an important role in inorganic phosphate (Pi) acquisition. Although the importance of cluster roots in Pi acquisition is well known, information on the distribution of APase within tissues of normal and cluster roots is lacking. Isoelectric focusing of APase isoforms as well as histochemical localization and visualization of APase were used to clarify the importance of secretory APase for P nutrition of white lupin grown under P deficiency. Isoelectric focusing revealed that both the secretory type and other major APase isoforms probably involved in P translocation were inducible. The major activity in the rhizosphere soil of cluster roots and roots grown under hydroponic conditions corresponded to LASAP2, a previously purified APase secreted from white lupin roots. Histochemical localization using enzyme-labeled fluorescence (ELF)-97 phosphate as a substrate was applied to rhizosphere samples. This substrate provides fluorescent precipitates after hydrolysis by phosphatase. Strong APase activity in the epidermal tissues of normal roots and cluster rootlets and in root hairs of cluster rootlets under P deficiency was detected. These results support the hypothesis that APase activities in the rhizosphere liberate Pi and supply it to white lupin plants grown under P-deficient conditions.

Effect of exogenous phosphatase and phytase activities on organic phosphate mobilization in soils with different phosphate adsorption capacities

This study evaluated the effects of exogenous LASAP2 for acid phosphatase (APase) and LASAP3 for phytase of white lupin (Lupinus albus L.) on phosphorus (P) accumulation from organic P in soils. The potential for LASAP2-overexpressing tobacco (Nicotiana tabacum L.) to increase organic P in soil was examined in our previous study. However, LASAP2 has low specificity for phytate, the predominant form of unavailable P in the brown lowland soil. For the present study, we isolated the full length of LASAP3 cDNA and introduced it into tobacco plants using Agrobacterium-mediated transformation. Transgenic tobacco plants were grown in two different soils (Andosols and Regosols; high and low P-adsorption capacity, respectively) supplemented with either inorganic phosphate (+Pi) or phytate (Po) as the sole P source, or control conditions that lacked phosphorus (No P). Dry matter production and P content of the transgenic line was higher than that of wild type in all treatments. The ratio of P accumulation increase by exogenous enzymes was found to be dependent on the P treatment and soil type. In all lines, the increase in +Po was less than that in +Pi, but higher than in No P. The P uptake ability of plants in Regosols was higher than in Andosols for all treatments, suggesting that the P utilization efficiency of both Pi and Po is dependent on the solubility. In no soil type or P treatment was a significant difference found between LASAP2- and LASAP3-overexpressing lines. These results demonstrate that introducing an APase and phytase gene such as LASAP2 and LASAP3 into tobacco by genetic transformation is a promising strategy for improving P mobilization in soil, although the bottleneck for mobilization of phytate-P is not the specificity of the enzyme but its solubility in soils.

Landrace of japonica rice, Akamai exhibits enhanced root growth and efficient leaf phosphorus remobilization in response to limited phosphorus availability

AimsPhosphorus (P) acquisition through extensive root growth and P allocation to different plant organs through efficient remobilization are important for acclimation of crop plants to P-limited environments. This study elucidated changes in rice root growth and leaf P-remobilization and their influence on grain yield under P deficiency.MethodsTwo pot experiments were conducted with (P100) and without (P0) inorganic P supply using two Japanese rice cultivars: Akamai (Yamagata) and Koshihikari. Multiple harvests were made until the panicle initiation stage. Root and shoot growth response, P acquisition, and temporal leaf P-remobilization efficiency were measured. A separate experiment ascertained the final yield and grain P status.ResultsThe Akamai rice cultivar showed enhanced root growth and more acquired soil P. The Akamai root dry weight was 66% greater than that of Koshihikari under P0. Confronting P deficiency, Akamai remobilized some P from its lower mature leaves to upper younger leaves starting from early growth. The remobilized P fraction increased to 72% at panicle initiation under P0. Under P0, Akamai exhibited two-fold higher leaf P-remobilization efficiency than under P100.ConclusionsEnhanced root growth that facilitates acquisition of more soil P through better soil exploration coupled with efficient leaf P remobilization from the early growth stage improves adaptation of Akamai rice cultivar to P-limited environments. Nevertheless, P-starvation responses did not facilitate higher grain yields in P-limited conditions.

Molecular Approaches to the Study of Biological Phosphorus Cycling

Phosphorus cycling in soils is partly affected by biological processes. Many different organisms are involved, and microbial functions in particular make a substantial contribution. Recently, various molecular tools independent of microbial cultivation have been developed, offering new possibilities for the analysis of the function and ecology of microbes involved in phosphorus cycling. Plants are also directly or indirectly involved in biological phosphorus cycling. Molecular approaches are powerful for understanding plant functions and the plant–microbe interactions involved in phosphorus cycling. In this chapter, the application of molecular tools to the study of the role of plants and rhizosphere microorganisms in phosphorus cycling is discussed.

Identification of genomic regions associated with low phosphorus tolerance in japonica rice (Oryza sativa L.) by QTL-Seq

ABSTRACT Phosphorus (P) is a major nutrient supporting rice productivity. Improving low-P tolerance of rice is expected to reduce dependence on P fertilizer, thereby reducing rice production costs and environmental impacts. This report describes the mapping of quantitative trait loci (QTL) associated with P deficiency tolerance in japonica rice. An F5 population derived from a cross of the low-P tolerant cultivar Akamai (Yamagata) and the sensitive cultivar Koshihikari was evaluated for shoot growth under low-P conditions. Then single nucleotide polymorphism (SNP) profiles of the low-P tolerant and sensitive bulks were compared on a genome-wide scale by QTL-Seq, a rapid QTL mapping method using next-generation sequencing technology. Results show a major QTL associated with low-P tolerance located on the long arm of chromosome 12. It has been named QTL for low-P tolerance 1 or qLPT1. SNPs were detected in 45 genes of qLPT1 region and the 5 genes were harboring synonymous SNPs, although none of them had been reported as involved in low-P tolerance. This result implies that the novel gene responsible for low-P tolerance exists in qLPT1. This study will contribute to the elucidation of mechanisms underlying low-P tolerance of Akamai and will facilitate the breeding of rice with low-P tolerance.

Ancient rice cultivar extensively replaces phospholipids with non-phosphorus glycolipid under phosphorus deficiency

Recycling of phosphorus (P) from P-containing metabolites is an adaptive strategy of plants to overcome soil P deficiency. This study was aimed at demonstrating differences in lipid remodelling between low-P-tolerant and -sensitive rice cultivars using lipidome profiling. The rice cultivars Akamai (low-P-tolerant) and Koshihikari (low-P-sensitive) were grown in a culture solution with [2 mg l-1 (+P)] or without (-P) phosphate for 21 and 28 days after transplantation. Upper and lower leaves were collected. Lipids were extracted from the leaves and their composition was analysed by liquid chromatography/mass spectrometry (LC-MS). Phospholipids, namely phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylglycerol (PG) and phosphatidylinositol (PI), lysophosphatidylcholine (lysoPC), diacylglycerol (DAG), triacylglycerol (TAG) and glycolipids, namely sulfoquinovosyl diacylglycerol (SQDG), digalactosyldiacylglycerol (DGDG), monogalactosyldiacylglycerol (MGDG) and 1,2-diacyl-3-O-alpha-glucuronosyl glycerol (GlcADG), were detected. GlcADG level was higher in both cultivars grown in -P than in +P and the increase was larger in Akamai than in Koshihikari. DGDG, MGDG and SQDG levels were higher in Akamai grown in -P than in +P and the increase was larger in the upper leaves than in the lower leaves. PC, PE, PG and PI levels were lower in both cultivars grown in -P than in +P and the decrease was larger in the lower leaves than in the upper leaves and in Akamai than in Koshihikari. Akamai catabolised more phospholipids in older leaves and synthesised glycolipids in younger leaves. These results suggested that extensive phospholipid replacement with non-phosphorus glycolipids is a mechanism underlying low-P-tolerance in rice cultivars.

Transgenic approaches for improving phosphorus use efficiency in plants

Abstract Phosphorus (P) is easily fixed in soils, forming unavailable forms, such as insoluble P and organic P. Plants frequently face P deficiency because of its low mobility in soil. Many strategies to adapt to P-deficient conditions have been developed in plants, but they can be categorized as two major strategies: (1) efficient use of sparingly available P by increase root exudation, including organic acids and phosphatases, by increase expression of phosphate transporter, and by modification of root architecture; and (2) improvement of internal P use efficiency by alteration of carbon metabolisms and by lipid remodeling. The strategies are regulated by a complex signaling network. This chapter presents an overview of the strategies and past trials to improve P use efficiency by transformation of certain genes involved in low-P stress response including future perspectives.

Dark conditions enhance aluminum tolerance in several rice cultivars via multiple modulations of membrane sterols

Aluminum tolerance of aluminum-sensitive rice was enhanced under darkness by multiple changes in membrane sterols: decreased stigmasterol, increased precursor partitioning for sterols biosynthesis, and increased expression of HMG genes.