Contribution of the grain size QTL GS3 to yield properties and physiological nitrogen-use efficiency in the large-grain rice cultivar 'Akita 63'.

The development of crop varieties with high nitrogen-use efficiency (NUE) is thought to be important in achieving sustainable cereal crop production. The high yield large-grain rice cultivar Oryza sativa L. 'Akita 63' (temperate japonica) has high physiological NUE (PNUE) for grain yield (GY). Our previous study revealed that a large-grain allele of GS3 is present in 'Akita 63'. Here, we verified the influence of GS3 on the yield properties and PNUE for GY in 'Akita 63'. The frequency distribution of brown rice length in F2 crosses of 'Iwate 75' and 'Akita 63' showed a continuous distribution that could be explained by GS3. A near-isogenic line was developed to substitute the GS3 segment of 'Koshihikari', which harbours a normal-sized grain allele, in the genetic background of 'Akita 63' and the line was designated as Akita63NILGS3-Koshihikari. Compared with Akita63NILGS3-Koshihikari, 'Akita 63' exhibited a significantly increased grain length, single brown grain weight and GY, although no significant differences were observed in the nitrogen content and above-ground biomass per unit of cultivated area. These results indicate that the GS3 large-grain allele is a contributing factor to high PNUE for GY in 'Akita 63'. These findings will facilitate the development of nitrogen-efficient rice varieties.

Breeding and characterization of the high cadmium-accumulating rice line 'Akita 119'.

Cadmium (Cd) is a toxic heavy metal that is mainly accumulated through the consumption of foods produced in Cd-contaminated fields. Phytoremediation is one of the most effective methods to reduce the soil Cd concentration. In this study, we bred a new rice line, 'Akita 119', for Cd phytoremediation. 'Akita 119' was obtained by a soft X-ray mutation of 'Cho-ko-koku', a naturally high-Cd-accumulating rice cultivar. The heading date of 'Akita 119' was about 2 weeks later than that of 'Akitakomachi', which is the leading cultivar in Akita Prefecture, Japan. 'Akita 119' has a short culm length and many panicles. The shattering resistance and lodging resistance of 'Akita 119' were improved compared to 'Cho-ko-koku'. The thousand-grain weight of 'Akita 119' was much smaller than that of 'Akitakomachi', and grains of 'Akita 119' could be easily distinguished from general japonica cultivars. When 'Akita 119' was grown in Cd-contaminated fields, the shoot dry weight and Cd concentration were similar to those of 'Cho-ko-koku'. These results demonstrate that 'Akita 119' has improved agronomic characteristics compared to 'Cho-ko-koku' while retaining the ability to extract Cd. Therefore, it should be considered a promising candidate for Cd phytoremediation in paddy fields in northern parts of Japan.

Mutations in rice (Oryza sativa) heavy metal ATPase 2 (OsHMA2) restrict the translocation of zinc and cadmium.

Widespread soil contamination with heavy metals has fostered the need for plant breeders to develop new crops that do not accumulate heavy metals. Metal-transporting transmembrane proteins that transport heavy metals across the plant plasma membrane are key targets for developing these new crops. Oryza sativa heavy metal ATPase 3 (OsHMA3) is known to be a useful gene for limiting cadmium (Cd) accumulation in rice. OsHMA2 is a close homolog of OsHMA3, but the function of OsHMA2 is unknown. To gain insight into the function of OsHMA2, we analyzed three Tos17 insertion mutants. The translocation ratios of zinc (Zn) and Cd were clearly lower in all mutants than in the wild type, suggesting that OsHMA2 is a major transporter of Zn and Cd from roots to shoots. By comparing each allele in the OsHMA2 protein structure and measuring the Cd translocation ratio, we identified the C-terminal region as essential for Cd translocation into shoots. Two alleles were identified as good material for breeding rice that does not contain Cd in the grain but does contain some Zn, and that grows normally.

OsHMA3, a P1B-type of ATPase affects root-to-shoot cadmium translocation in rice by mediating efflux into vacuoles.

• The cadmium (Cd) over-accumulating rice (Oryza sativa) cv Cho-Ko-Koku was previously shown to have an enhanced rate of root-to-shoot Cd translocation. This trait is controlled by a single recessive allele located at qCdT7. • In this study, using positional cloning and transgenic strategies, heavy metal ATPase 3 (OsHMA3) was identified as the gene that controls root-to-shoot Cd translocation rates. The subcellular localization and Cd-transporting activity of the gene products were also investigated. • The allele of OsHMA3 that confers high root-to-shoot Cd translocation rates (OsHMA3mc) encodes a defective P(1B) -ATPase transporter. OsHMA3 fused to green fluorescent protein was localized to vacuolar membranes in plants and yeast. An OsHMA3 transgene complemented Cd sensitivity in a yeast mutant that lacks the ability to transport Cd into vacuoles. By contrast, OsHMA3mc did not complement the Cd sensitivity of this yeast mutant, indicating that the OsHMA3mc transport function was lost. • We propose that the root cell cytoplasm of Cd-overaccumulating rice plants has more Cd available for loading into the xylem as a result of the lack of OsHMA3-mediated transportation of Cd to the vacuoles. This defect results in Cd translocation to the shoots in higher concentrations. These data demonstrate the importance of vacuolar sequestration for Cd accumulation in rice.

The Road to Practical Application of Cadmium Phytoremediation Using Rice.

Cadmium (Cd) is a toxic heavy metal that causes severe health issues in humans. Cd accumulates in the human body when foods produced in Cd-contaminated fields are eaten. Therefore, soil remediation of contaminated fields is necessary to provide safe foods. Rice is one of the primary candidates for phytoremediation. There is a genotypic variation of Cd concentration in the shoots and grains of rice. Using the world rice core collection, 'Jarjan', 'Anjana Dhan', and 'Cho-ko-koku' were observed with a significantly higher level of Cd accumulation in the shoots and grains. Moreover, OsHMA3, a heavy metal transporter, was identified as a responsive gene of quantitative trait locus (QTL) for high Cd concentration in the shoots of these three varieties likewise. However, it is difficult to apply practical phytoremediation to these varieties because of their unfavorable agricultural traits, such as shatter and easily lodged. New rice varieties and lines were bred for Cd phytoremediation using OsHMA3 as a DNA marker selection. All of them accumulated Cd in the shoots equal to or higher than 'Cho-ko-koku' with improved cultivation traits. Therefore, they can be used for practical Cd phytoremediation.

A single recessive gene controls cadmium translocation in the cadmium hyperaccumulating rice cultivar Cho-Ko-Koku.

The heavy metal cadmium (Cd) is highly toxic to humans and can enter food chains from contaminated crop fields. Understanding the molecular mechanisms of Cd accumulation in crop species will aid production of safe Cd-free food. Here, we identified a single recessive gene that allowed higher Cd translocation in rice, and also determined the chromosomal location of the gene. The Cd hyperaccumulator rice variety Cho-Ko-Koku showed 3.5-fold greater Cd translocation than the no-accumulating variety Akita 63 under hydroponics. Analysis of an F(2) population derived from these cultivars gave a 1:3 segregation ratio for high:low Cd translocation. This indicates that a single recessive gene controls the high Cd translocation phenotype. A QTL analysis identified a single QTL, qCdT7, located on chromosome 7. On a Cd-contaminated field, Cd accumulation in the F(2) population showed continuous variation with considerable transgression. Three QTLs for Cd accumulation were identified and the peak of the most effective QTL mapped to the same region as qCdT7. Our data indicate that Cd translocation mediated by the gene on qCdT7 plays an important role in Cd accumulation on contaminated soil.