Yumiko Arai-Sanoh

A novel lime pretreatment for subsequent bioethanol production from rice straw--calcium capturing by carbonation (CaCCO) process.

In order to establish an efficient bioethanol production system for rice straw, we developed a novel lime-pretreatment process (CaCCO process) that did not require a solid-liquid-separation step. This process adopted a step in which after pretreatment lime was neutralized by carbonation, resulting in a final pH of about 6. CaCO(3) produced by the process was kept in the reaction vessel, and no significant inhibitory effects on enzymatic saccharification and fermentation were observed. In the CaCCO process, solubilized carbohydrates, such as xylan, starch, and sucrose were also kept in the vessel, enabling high recoveries of monomeric sugars. Simultaneous saccharification and fermentation (SSF) of pretreated rice straw, 10% (g-rice straw/g-water), using Saccharomyces cerevisiae and Pichia stipitis yielded 19.1 g L(-1) ethanol that was 74% of the theoretical yield from glucose and xylose. Thus, this process represents a novel pretreatment method to utilize not only cellulose but also xylan, starch, and sucrose from biomass.

Efficient Recovery of Glucose and Fructose via Enzymatic Saccharification of Rice Straw with Soft Carbohydrates

Soft carbohydrates, defined as readily-recoverable carbohydrates via mere extraction from the biomass or brief enzymatic saccharification, were found in significant amounts in rice straw as forms of free glucose, free fructose, sucrose, starch, and β-1,3-1,4-glucan. In this study, we investigated their amounts in rice straw (defined as culm and leaf sheath), and developed an easy method for glucose and fructose recovery from them with heat-pretreatment and subsequent 4-h enzymatic saccharification with an enzyme cocktail of cellulase and amyloglucosidase. The recovery of glucose and fructose exhibited good correlation with the amounts of soft carbohydrates. The maximum yields of glucose and fructose in the rice straw per dry weight at the heading stage and the mature stage were 43.5% in cv. Habataki and 34.1% in cv. Leafstar. Thus, rice straw with soft carbohydrates can be regarded as a novel feedstock for economically feasible production of readily-fermentable glucose and fructose for bioethanol.

Deep rooting conferred by DEEPER ROOTING 1 enhances rice yield in paddy fields

To clarify the effect of deep rooting on grain yield in rice (Oryza sativa L.) in an irrigated paddy field with or without fertilizer, we used the shallow-rooting IR64 and the deep-rooting Dro1-NIL (a near-isogenic line homozygous for the Kinandang Patong allele of DEEPER ROOTING 1 (DRO1) in the IR64 genetic background). Although total root length was similar in both lines, more roots were distributed within the lower soil layer of the paddy field in Dro1-NIL than in IR64, irrespective of fertilizer treatment. At maturity, Dro1-NIL showed approximately 10% higher grain yield than IR64, irrespective of fertilizer treatment. Higher grain yield of Dro1-NIL was mainly due to the increased 1000-kernel weight and increased percentage of ripened grains, which resulted in a higher harvest index. After heading, the uptake of nitrogen from soil and leaf nitrogen concentration were higher in Dro1-NIL than in IR64. At the mid-grain-filling stage, Dro1-NIL maintained higher cytokinin fluxes from roots to shoots than IR64. These results suggest that deep rooting by DRO1 enhances nitrogen uptake and cytokinin fluxes at late stages, resulting in better grain filling in Dro1-NIL in a paddy field in this study.

Genetic mechanisms underlying yield potential in the rice high-yielding cultivar Takanari, based on reciprocal chromosome segment substitution lines

BackgroundIncreasing rice yield potential is a major objective in rice breeding programs, given the need for meeting the demands of population growth, especially in Asia. Genetic analysis using genomic information and high-yielding cultivars can facilitate understanding of the genetic mechanisms underlying rice yield potential. Chromosome segment substitution lines (CSSLs) are a powerful tool for the detection and precise mapping of quantitative trait loci (QTLs) that have both large and small effects. In addition, reciprocal CSSLs developed in both parental cultivar backgrounds may be appropriate for evaluating gene activity, as a single factor or in epistatic interactions.ResultsWe developed reciprocal CSSLs derived from a cross between Takanari (one of the most productive indica cultivars) and a leading japonica cultivar, Koshihikari; both the cultivars were developed in Japan. Forty-one CSSLs covered most of the Takanari genome in the Koshihikari background and 39 CSSLs covered the Koshihikari genome in the Takanari background. Using the reciprocal CSSLs, we conducted yield trials under canopy conditions in paddy fields. While no CSSLs significantly exceeded the recurrent parent cultivar in yield, genetic analysis detected 48 and 47 QTLs for yield and its components in the Koshihikari and Takanari backgrounds, respectively. A number of QTLs showed a trade-off, in which the allele with increased sink-size traits (spikelet number per panicle or per square meter) was associated with decreased ripening percentage or 1000-grain weight. These results indicate that increased sink size is not sufficient to increase rice yield in both backgrounds. In addition, most QTLs were detected in either one of the two genetic backgrounds, suggesting that these loci may be under epistatic control with other gene(s).ConclusionsWe demonstrated that the reciprocal CSSLs are a useful tool for understanding the genetic mechanisms underlying yield potential in the high-yielding rice cultivar Takanari. Our results suggest that sink-size QTLs in combination with QTLs for source strength or translocation capacity, as well as careful attention to epistatic interactions, are necessary for increasing rice yield. Thus, our findings provide a foundation for developing rice cultivars with higher yield potential in future breeding programs.

Effects of Soil Temperature on Growth and Root Function in Rice

Abstract The objective of this study was to clarify the effects of soil temperature in the stage from late tillering to panicle initiation (SI) and during the grain-filling stage (SII) on grain setting, dry matter production, photosynthesis, non-structural carbohydrate (NSC), xylem exudation and abscisic acid (ABA) levels in rice (Oryza sativa L. cv. Koshihikari). Rice plants were exposed to four different soil temperatures during SI or SII: 17.5, 25, 31.5 and 36.5°C (ST18, ST25, ST32 and ST37, respectively). The yield, yield components, grain filling and quality in SI were negatively influenced by high soil temperature of 37°C. On the other hand, there was no significant difference in those characters among temperature treatments in SII. The root/shoot ratio was smallest in the ST37 plants in both SI and SII, mainly due to their lighter root weight. At 7 days after initiation of treatment (DAT) in both SI and SII, the photosynthetic and xylem exudation rate tended to increase slightly as soil temperature increased up to 32°C. At 21 DAT, however, the photosynthetic rate was lowest in ST37, with concurrent decrease of diffusion conductance and SPAD value. In addition, decrease of NSC concentration in stem and xylem exudation rate, and increase of ABA level in leaves and xylem exudate were observed in ST37 plants at 21 DAT. These results suggested that high soil temperature before heading especially influenced yield, grain quality and plant growth. Possible mechanisms of the effect of soil temperature are discussed.

Genotypic Variations in Non-Structural Carbohydrate and Cell-Wall Components of the Stem in Rice, Sorghum, and Sugar Vane

We evaluated genetic variations in the non-structural carbohydrate (NSC) and the cell-wall components of stem in rice, sorghum, and sugar cane to assess the potential suitability of these gramineous crops for bioethanol production. For NSC, the maximum soluble sugar concentration was highest in sugar cane, followed by sorghum with sucrose. The major NSC in rice was starch, but there were wide variations in the starch to soluble sugar ratios among the cultivars. The total concentration of cell-wall components was negatively correlated with the NSC concentration, indicating competition for carbon sources. Among the cell-wall components, lignin was relatively stable within each group. The major sugar species composing hemicellulose was xylose in all crop groups, but there were differences in composition, with a higher fraction of arabinose and glucose in rice as compared to the other crops. In rice, there was less lignin than in sorghum or sugar cane; this might be advantageous for the efficient saccharification of cellulose.

Comparative analysis of the relationship between Cs and K in soil and plant parts toward control of Cs accumulation in rice

Abstract The effect of soil exchangeable potassium (K) and cesium (Cs) levels on Cs uptake and accumulation in different parts of rice (Oryza sativa L.) plants were examined using paddy soils with diverse exchangeable K and Cs in pot experiments. Aboveground Cs uptake decreased with higher exchangeable K and was linearly correlated with exchangeable Cs/K ratios, indicating competitive absorption of these elements by roots. Variation in Cs concentration in brown rice among soils was also related to the exchangeable Cs/K ratio. The exchangeable Cs/K ratio was positively reflected in the Cs/K concentration ratio in each plant part, with a specific slope, suggesting that Cs transport was coordinated with K transport and that there were regulated discriminations of Cs against K in the translocation process among parts. The Cs/K ratio was higher in brown rice and dead leaves than in active leaves, stems and husks. The distribution of Cs accumulation in brown rice was 14.5% on average, but it was variable and negatively related to K concentration in the stem. The Cs distribution in aboveground plant parts also decreased with higher K concentration in the root. These results imply the importance of the competitiveness with K in the root absorption and translocation of Cs within the plant. Based on the observed relationship between Cs and K, effective K management and other measures to control Cs accumulation in plant parts are discussed.

Exchangeable Cs/K ratio in soil is an index to estimate accumulation of radioactive and stable Cs in rice plant

Abstract Pot and field experiments were conducted to clarify the effect of soil exchangeable potassium (K) and cesium-137 (137Cs) on 137Cs accumulation and to establish soil index in rice (Oryza sativa L.). Four paddy soils in Fukushima Prefecture, Japan, showing different transfer factors for radioactive Cs derived from the accident of Fukushima Daiichi Nuclear Power Station in the field were compared in terms of 137Cs accumulation in rice in a pot experiment. 137Cs accumulation in shoots and brown rice widely varied among soils with the transfer factor ranging from 0.018 to 0.068 for shoots and 0.004 to 0.065 for brown rice. 137Cs concentration in brown rice and shoots tended to decrease with higher levels of soil exchangeable K, and they were more closely related to the exchangeable Cs/K ratio. Similar relationships between the Cs/K ratio and Cs accumulation in plants were obtained for the stable isotope cesium-133 (133Cs). The distributions of 137Cs and 133Cs in grains were also similar and variable among soils. The transfer factors obtained in pot experiments mostly agreed with field observations. The results imply that the exchangeable 137Cs/K can be a potential soil index to estimate 137Cs accumulation in rice.

Identification of Chromosomal Regions Controlling the Leaf Photosynthetic Rate in Rice by Using a Progeny from Japonica and High-yielding Indica Varieties

Abstract The whole-leaf photosynthetic rate in rice plants is controlled by various physiological processes. In a high-yielding indica rice variety, Habataki, the leaf photosynthetic rate (LPR) of the uppermost fully expanded leaves was approximately 130 to 140% of that in a japonica variety, Sasanishiki, from booting to the early ripening stage. We characterized the difference in the LPR between Habataki and Sasanishiki. Leaves of Habataki contained higher levels of nitrogen and, as a consequence, of Rubisco, and had higher stomatal conductance that was associated with higher hydraulic conductance from roots to leaves than those of Sasanishiki. These features were responsible for the higher LPR of Habataki. An analysis of chromosome segment substitution lines (CSSLs) in which chromosome segments from Habataki were substituted into the genetic background of Sasanishiki showed that three genetic regions on chromosomes 4, 5 and 11 were responsible for the increase in the LPR. Each of these regions was estimated to increase the LPR by 15 to 30%, and we showed that they were associated with higher activity of mesophyll photosynthesis due to higher leaf nitrogen content and greater stomatal conductance. Leaf nitrogen content and stomatal conductance may be useful parameters for further quantitative trait locus analysis of efficient photosynthesis in leaves.

DiSC (direct saccharification of culms) process for bioethanol production from rice straw.

A simple process (the direct-saccharification-of-culms (DiSC) process) to produce ethanol from rice straw culms, accumulating significant amounts of soft carbohydrates (SCs: glucose, fructose, sucrose, starch and β-1,3-1,4-glucan) was developed. This study focused on fully mature culms of cv. Leafstar, containing 69.2% (w/w of dried culms) hexoses from SCs and cellulose. Commercially-available wind-separation equipment successfully prepared a culm-rich fraction with a SC recovery of 83.1% (w/w) from rice straw flakes (54.1% of total weight of rice straw). The fraction was suspended in water (20%, w/w) for starch liquefaction, and the suspension was subjected to a simultaneous saccharification and fermentation with yeast, yielding 5.6% (w/v) ethanol (86% of the theoretical yield from whole hexoses in the fraction) after 24h fermentation. Thus, the DiSC process produced highly-concentrated ethanol from rice straw in a one vat process without any harsh thermo-chemical pretreatments.