Legumes are mostly grown rainfed and are exposed to various types of drought ranging from terminal drought to intermittent drought. The objective of this study was to compare the root anatomy of six major legume crops in relation to their drought adaptation strategies. Plants of chickpea (Cicer arietinum L.), groundnut (Arachis hypogaea L.), pigeonpea (Cajanus cajan [L.] Millsp.), cowpea (Vigna unguiculata L. walp.), soybean (Glycine max [L.] Merr.) and common bean (Phaseolus vulgaris L.) were grown along with pearl millet (Pennisetum glaucum [L.] R. Br.) in a Vertisol field during the rainy season of 2010. Four root segments from 35-day-old plants of each crop species were collected, 10 cm from the root tip and used for making transverse sections. These root segments were thinner in both groundnut and pigeonpea than in other legumes but similar to those of pearl millet. Soybean and pigeonpea had a relatively thinner cortex than the other legumes. Xylem vessel size and the numbers were apparently the most discriminating traits of legumes. Pigeonpea is equipped to conduct small quantities of water per unit time with a few narrow xylem vessels and that explains the conservative early growth of pigeonpea. Chickpea and cowpea showed moderate xylem passage per root indicating that they are capable of absorbing water moderately and are well equipped for regular drought episodes. The development of cortical and stele tissue and their proportion is markedly influenced by moisture availability to the root system.

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Digital images for determining root traits have been analyzed using WinRHIZO, commercial software. A freeware ImageJ has been developed for the analysis but whether root lengths assigned to root diameter classes obtained by this method correspond to those estimated by WinRHIZO has not been confirmed. In this study, root lengths estimated using ImageJ were compared with those estimated using WinRHIZO. In ImageJ, the root lengths were estimated using a triangle thresholding algorithm that was valid for total root length in our previous study. For the small-diameter classes (< 0.5 mm), the estimates obtained using ImageJ and the conversion factor 2/3, corresponded closely to the estimates obtained using WinRHIZO. For large-diameter classes (> 0.5 mm), root lengths estimated using ImageJ were similar to those obtained using WinRHIZO without using the conversion factor. These results were valid for rice roots. In this report, we propose completely automated estimation of root lengths for roots in each diameter class by using ImageJ.

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An appropriate combination of rice cultivar and cropping system that maximizes water use efficiency (WUE) may improve yield of rainfed lowland rice. In the paddy field, a large amount of water is consumed by evaporation during the early growth period, and it can be reduced by canopy coverage especially in semi-arid regions. Therefore, we evaluated the role of canopy coverage in WUE of rice in the early growth period in semi-arid region. A pot experiment was conducted in Namibia to investigate the genotypic and species difference in WUE, and another pot and a field experiment were conducted to investigate the effects of planting density on WUE. Although no significant difference was observed among species, the mean WUE was in the decreasing order of Oryza sativa, and Oryza glaberrima followed by the interspecific progenies including NERICA. In contrast, there was a significant difference in WUE at the genotypic level. Highly tillering genotypes such as WAB1159-2-12-11-5-1 and WITA 2 showed a high WUE. Furthermore, WUE was significantly correlated with the number of tillers (R²=0.453), and higher planting density resulted in a higher WUE. In contrast, stomatal conductance had no significant correlation with WUE (R²=0.081). Thus, the physical conditions affected by number of tillers and planting density had greater impacts on WUE than physiological characteristics such as stomatal conductance. The suppression of surface water evaporation by coverage was significant, probably contributing to WUE improvement. To increase WUE in semi-arid regions, we recommend the increase of canopy coverage and higher planting density.

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To evaluate the effect of inclination angle of the leaf on the leaf injury of rice by O₃, we examined the effect of a 5-hr exposure to 0, 0.1, 0.3 cm³ m^-3 O₃, abbreviated as O⁰, O^0.1 and O^0.3, respectively, of the flag leaf using natural-light gas-exposure chambers. The middle portion of the flag leaf was set horizontally using lead weights (H plant) and it was compared with a non-treated, erect leaf (E plant). Gas exchange and chlorophyll fluorescence were measured immediately before, immediately after, and 1 and 3 d after the start of the exposure. The net photosynthetic rate, stomatal conductance and operating quantum efficiency of photosystem II (F_q’/F_m’) were decreased by the exposure to O₃. Furthermore, the inhibitory effects of O^0.1 and O^0.3 on the H plants were severer than those on the E plants. The maximum quantum efficiency of photosystem II (F_v/F_m) in the leaf of H plants decreased immediately after exposure to O₃. These results indicate that the weaker inhibitory effect of O₃ on the erect leaf depends on the lower light intensity at the leaf surface, rather than the horizontal leaf position.

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The aim of this study was to obtain the information for improvement in source ability via expansion of a flag leaf area in rice (Oryza sativa L.). We identified chromosome regions affecting a trait (CRATs) for leaf area and confirmed their effects with two sets of chromosome segment substitution lines developed from indica landraces Kasalath or Nona Bokra as a donor under japonica cultivar Koshihikari genetic background. We detected 4 and 8 CRATs that increased leaf area by Kasalath and Nona Bokra alleles, respectively. With the Nona Bokra allele, CRAT on chromosome 7 (LA7) increased leaf area by 54% comparing to Koshihikari. LA7 maintained the contents of Rubisco and photosynthetic rate and as a result its “estimated” source ability of a flag leaf was increased by 23%. The effect of LA7 was stable under different environmental conditions, and might be useful for breeding rice with higher source ability and yield.

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Five ribulose-1,5-bisphosphate carboxylase/oxygenase small subunit genes identified in the rice (Oryza sativa) genome are designated as OsRBCS1, 2, 3, 4 and 5. OsRBCS1 transcripts were not detectable. The mRNA levels of OsRBCS2, OsRBCS3, OsRBCS4 and OsRBCS5 in leaf blades were increased by NH₄NO₃ supply to nitrogen-starved plants, but the extent of increment varied with the genes. The mRNA levels of OsRBCS2, OsRBCS3 and OsRBCS4 in leaf blades were increased by NH₄NO₃ or glutamine supply to detached shoots. On the other hand, NH₄NO₃ and glutamine had little effect on the expression of OsRBCS5 gene. The mRNA level of OsRBCS4 was increased only by cytokinin supply to the detached shoots. The expression of the promoters of OsRBCS2 and OsRBCS3 was enhanced by NH₄NO₃ supply.

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We investigated 321 varieties to identify the aleurone traits that are closely related to the lipid content of rice. Brown rice seeds were cut crosswise near the center with a razor blade and the cut surface was stained with Oil Red O, and then observed under integrated fluorescence microscope (BZ9000). We found wide variations among varieties in the area stained, but the japonica group contained many varieties with a large stained area. We selected 17 representative varieties covering the whole range of stained areas and confirmed the thickness of aleurone layer using Kawamoto’s film method (micro-thin sections that were created with cryomicrotome). Aleurone traits were examined under a light microscope and measured. We found a strong correlation between the area of the aleurone layer and the stained area (r = 0.799***) and between the thickness of the aleurone layer and the thickness of the stained region (r = 0.543*). The area of the stained region therefore provides a fast and effective indicator for selecting varieties for the thickness of the aleurone layer. We also measured the amount (mg per 100 seeds) and proportion of triacylglycerols (TAGs) to 1 g of seed (mg g^-1) in brown rice seeds without embryos. The amount and proportion of TAG to 1 g of seed were both significantly correlated with the area, average thickness and percentage of aleurone layer. This suggests that the aleurone traits will be good indicators for the selection of varieties with high levels of TAG.

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Direct seeding is a labor-saving rice cultivation method, but poor establishment is a major obstacle to the widespread use of this practice. This study examined the suppressive effect of ammonium sulfate, a nitrogen fertilizer, on seedling establishment in submerged soil. Establishment was impaired even at 20ºC by the application of sulfate salts, including ammonium sulfate and potassium sulfate, but not by ammonium chloride. Impairment by ammonium sulfate did not occur in autoclaved soil. In addition, the redox potential in the soil near the seeds decreased rapidly to levels that allowed sulfide ion generation, even at 20ºC. The sulfur concentration in the soil near the seeds decreased with the decline in redox potential, suggesting that sulfide ions were transformed from soluble sulfate ions into insoluble iron sulfide through binding with ferrous ions near the seeds. These results imply that sulfate ions may be transformed into sulfide ions by bacteria only around the seeds if sufficient sulfate ions are present in the soil, thereby impairing the establishment of seeds sown directly in submerged soil, even under cool springtime conditions and in paddy fields that do not dry out prior to submergence.

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Sulfide ion generation in strictly reduced soil might be a factor impairing rice seedling establishment. The molybdate ion is known to suppress the microbial transformation of sulfide ions from sulfate ions. I investigated the effect of molybdate on rice establishment in sulfate-applied submerged soil. The establishment of rice seeds sown in sulfate-applied submerged soil was markedly improved by application of potassium molybdate at a rate of 1.5−15 mmol kg^-1 dried soil. The application of potassium molybdate delayed the decline of the sulfur concentration in the soil solution near the seeds and the appearance of circular black stains, or insoluble iron sulfide indicating the generation of sulfide ion, around seeds in the soil. Irrespective of the application of molybdate, the redox potential near the seeds was low enough to allow generation of sulfide ions, implying that molybdate suppresses the generation of sulfide ions with no effect on redox potential. These results suggest that the application of molybdate could improve rice seedling establishment in sulfur-rich submerged soil by suppressing the generation of sulfide ions, that is a possible factor suppressing the establishment of rice seeds sown in sulfur-rich submerged soil.

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Excess phosphorus (P) has accumulated in Japanese paddy soils, due to fertilizer P inputs over crop requirement for several decades, and improvement of the excess of P is necessary in view of environmental conservation. This study aimed to evaluate the input/output balance of P related to soil P status in paddy rice systems, and to obtain a practical indication. Irrigated rice (Oryza sative L.) was cultivated on a gley soil from 1997 to 2006. Six field plots fertilized with commercial fertilizer, animal waste composts, green manure and none were included. Phosphorus input varied among plots from 0 to 73 kg ha^-1 yr^-1. Rice P uptake was approximately 20 kg ha^-1, indicating no response to the P input. This was attributed to a large amount of plant-available Bray- and Truog-P in our soils. In our fields, paddy rice could be cultivated with no P-containing fertilizer or amendment. As a result, increase in the P input led to an increase in partial P balance (PPB). Cumulative increase in PPB resulted in the increase in soil total P, whereas cumulative decrease of PPB tended to decrease it. Excess accumulation of the soil P results in a loss of P into the environment. We concluded that P fertilization should be restricted to 20 kg ha^-1 yr^-1 (corresponding to 46 kg ha^-1 yr^-1 as P₂O₅), based on evenly balanced P input with the rice P uptake. It is also important to include all of the P-containing fertilizers and amendments when determining the amount of application.

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Rapid development after emergence is important for seedling establishment and early vegetative growth, especially at a low planting density or inferior environmental conditions. This study was conducted to understand the varietal difference in the growth parameters during the seedling stage in soybean. Twenty-seven soybean varieties originating from six countries were examined in 2009 and 2010. The pots were arranged in a completely random block design with 5 replications (10 pots per variety), and the seedlings were sampled at 14 and 28 days after sowing (DAS). The shoot dry weight at 14 and 28 DAS was highly correlated with seed size, cotyledon digestion, and leaf area. However, no positive correlation was found between shoot dry weight and photosynthetic rate at 28 DAS. Chamame, a Japanese cultivar, with the largest seed size grew rapidly, and showed the heaviest shoot dry weight, greatest cotyledon digestion, fast leaf expansion and high photosynthetic rate. However, Moyashimame, a medium-seed-size cultivar, also grew rapidly with a high photosynthetic rate. Some varieties such as Tachinagaha (Japan), Hefeng (China), Parana and Pérola (Brazil), had a large or medium seed size, and high photosynthetic rate but showed a relatively small leaf area and light shoot dry weight. These results suggested that big seeds with rapid cotyledon digestion developed a wider leaf area and therefore large dry matter production, indicating that the conversion of stored energy was more important than the leaf photosynthetic activity for early growth.

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One of the limiting factors of soybean yield in southwestern Japan is soil moisture fluctuation from flooding to drought. The Farm-Oriented Enhancing Aquatic System (FOEAS), which consists of underdrain and sub-irrigation systems and can keep groundwater levels stable, was developed in 2005 to solve this problem. To obtain basic information on soybean cultivation using the FOEAS in southwestern Japan, we examined the growth response to groundwater level by using two cultivars (cv. Fukuyutaka and Sachiyutaka) in 2008 and 2010. Plant growth, yield, yield components and seed components (protein and oil contents) were analyzed. We also studied the effect of sowing time (normal or late). The groundwater level was controlled at depths of 20 and 35 cm or not treated (NT) as a control. The groundwater level could be kept at the target depths (20 and 35 cm) by FOEAS throughout the experimental period, but that in the NT plot fluctuated greatly. The growth and yield of Sachiyutaka might be increased by FOEAS, especially in late sowing, but the yield of normal sown Sachiyutaka was decreased in the 20-cm plot in 2010. Yield of late-sown Fukuyutaka tended to be decreased in the 20-cm plot. These results suggest that (1) the optimum groundwater level for yield of Sachiyutaka is between 20 and 35 cm, regardless of sowing time and (2) the groundwater level should not be kept at 20 cm for late-sown Fukuyutaka. Further studies are needed to determine the optimum groundwater level for each cultivar.

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In Japan, soybean is usually cultivated in fields that have been converted from rice paddies, and poor seedling establishment due to pre-emergence seedling damping-off is often observed during the rainy season. In this study, the factors that cause the damping-off in flooded soil were investigated under high soil moisture conditions in a greenhouse and in agricultural fields. In sterilized soil sampled from a soybean field, seedlings emerged well under 48-hr flooded conditions. In unsterilized soil, soybean seeds treated with the fungicide, mancozeb+metalaxyl exhibited much higher emergence rates even under 48-hr flooded conditions than the seeds treated with oxytetracycline+streptomycin, benomyl, or flutolanil. Pythium, Phytophthora, Mucorales, Trichoderma, Geotrichum-like microorganisms, and some fungi producing conidia in a false head, were isolated from decayed seedlings. Of the isolated microorganisms, oomycete microorganisms, Pythium helicoides, other Pythium sp., and Phytophthora sp. were pathogenic to soybeans under flooded conditions. As the length of the flooding period increased, pre-emergence seedling rot also increased. However, the pathogenic oomycetes rarely caused pre-emergence seedling rot under conditions without flooding. Furthermore, under flooded conditions, the damage caused by these pathogens was reduced by treating the seeds with mancozeb+metalaxyl. These results indicate that oomycete microorganisms are involved in pre-emergence seedling damping-off under flooded soil conditions.

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