Distribution and characterization of enzymes causing starch degradation in rice (Oryza sativa cv. koshihikari)

The thermal dependency and stability of enzymes producing reducing sugar (RS) were examined in bran, the exterior 13% part (outer endosperm), and the remaining inner endosperm of rice grains. RS-producing enzymes in the inner endosperm showed a higher optimum temperature than those in other parts of the rice grain. Diethylaminoethyl-Sephacel chromatography of crude extracts revealed two peaks of RS-producing activity with different optimum temperatures (60 and 37 degrees C) in all three parts. alpha-Glucosidase (EC 3.2.1.20) and alpha-amylase (EC 3.2.1.1) isoform G were thought to be major components of the RS-producing activities with high and low optimum temperatures, respectively. The peak with a high optimum temperature was a more abundant component in the inner endosperm, compared with other parts of the rice grain. Thus, different parts of rice were found to have distinct enzyme sets having different thermal dependency and to be involved in starch degradation to various sugars.     

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.     

Effect of seed phosphorus and soil phosphorus applications on early growth of rice (Oryza sativa L.) cv. IR66

Abstract

The vigour and size of rice seedlings in the nursery are generally correlated with final grain yield. The present study examined the possibility that increasing seed phosphorus (P) concentration would stimulate early growth of rice seedlings and therefore would have the potential to increase rice yield. Rice seeds with a uniform size and three levels of P concentration (0.115, 0.173, and 0.240% on a dry weight basis) were sown in pots on a P deficient soil with three levels of P supply (0, 7.75, and 38.8 mg P kg^?1 soil) to investigate their effect on root and shoot dry weight and P accumulation at three harvest times, 10, 20, and 30 d after sowing (DAS). The effect of seed P concentration on plant growth was greatest at a low soil P concentration and it was less pronounced with increasing soil P concentration and with time at all levels of soil P. At 10 DAS, shoot dry weight was 15% higher at a high seed P concentration (0.240%) (p < O.O1) than at a low seed P concentration (0.115%) at each level of soil P supply whereas at subsequent harvests (20 and 30 DAS) the effect of seed P concentration was observed only when the soil P supply was deficient. In contrast with its effects on shoot dry weight, high seed P concentration increased root dry weight only at the latest harvest (30 DAS). The fact that high seed P increased P concentrations in shoot tips, and in roots at 10 DAS suggests that improved P nutrition of seedlings in the first 10 DAS may be the mechanism by which high seed P concentration stimulates early growth, especially in soils with low P concentration. Sowing rice seed with high P concentration may be beneficial for increasing farmer's rice yields, in P deficient soil, and requires further field investigations.     

Nitrogen balance in a paddy field planted with whole crop rice (Oryza sativa cv. Kusahonami) during two rice-growing seasons

This paper focuses on N balance in a paddy field planted with whole crop rice (Oryza sativa cv. Kusahonami). The experiment was conducted with two treatments during two rice-growing seasons: one was fertilized with N (160 kg N ha^–1; 16N plot) and the other unfertilized (0N plot); both plots were fertilized with P and K. The N input from precipitation was 15 and 12 kg N ha^–1 in 2002 and 2003, respectively. The N input from irrigation water reached as much as 123 and 69 kg N ha^–1 in 2002 and 2003, respectively. This was because irrigation water contained higher NO₃^– concentrations ranging from 4 to 8 mg N l^–1. The N uptake by rice plants was the major output: 118 and 240 kg N ha^–1 in the 0N and 16N plots in 2002 and 103 and 238 kg N ha^–1 in 2003, respectively. N losses by leaching were 4.8–5.3 and 6.5–7.3 kg N ha^–1 in 2002 and in 2003, respectively. Laboratory experiments were carried out to estimate the amounts of N₂ fixation and denitrification. Amount of N₂ fixation was 43 and 0 kg N ha^–1 in the 0N and 16N plots, respectively. Denitrification potential was quite high in both the plots, and 90% of the N input through irrigation water was lost through denitrification. Collectively, the total N inputs were relatively large due to irrigation water contaminated with NO₃^–, but N outflow loading, expressed as leaching–(irrigation water + precipitation + fertilizer), showed large negative values, suggesting that the whole crop rice field might serve as a constructed wetland for decreasing N.     

Identification of genomic locations associated with grain micronutrients (iron and zinc) in rice (Oryza sativa L.)

Genetic Resources and Crop Evolution - In this investigation, recombinant inbred lines (RILs) population derived from PAU 201 (high yielding) and Palman 579 (high iron and zinc content) varieties...     

Foliar-applied trehalose modulates growth,mineral nutrition,photosynthetic ability,and oxidative defense system of rice (Oryza sativa L.) under saline stress

A tub experiment was conducted to assess the effect of exogenously applied trehalose (0, 10, and 20 mM) on various attributes of two rice cultivars (Bas-385 and Bas-2000) under salt stress (0, 50, 100, and 150 mM). Salinity decreased growth, gas exchange characteristics, shoot and root potassium (K⁺) ions, hydrogen peroxide (H₂O₂), total soluble proteins, activity of catalase (CAT), and yield attributes, while it increased chlorophyll contents, shoot and root sodium (Na⁺) and calcium (Ca²⁺), malondialdehyde (MDA), glycinebetain (GB), free proline, and peroxidase (POD) activity. Foliar-applied trehalose improved growth attributes, net photosynthetic rate, GB, total soluble proteins, superoxide dismutase (SOD) and yield. Yield was not obtained at 150 mM salt stress. The rice cultivar Bas-2000 showed better performance with respect to gas exchange attributes and activities of enzymatic antioxidants. Overall, varying levels of foliar-applied trehalose proved to be effective in ameliorating adverse effects of salt stress on rice.     

Molecular cloning and characterization of oryzacystatin-III, a novel member of phytocystatin in rice (Oryza sativa L. japonica)

On the basis of cDNA sequences, we found that the calli of rice encodes an amino acid sequence that shares 56% and 89% identity, respectively, with oryzacystatin-I and oryzacystatin-II. This sequence differs from that of oryzacystatin-II in the N-terminal region (Gln(7)-Ala(19) in the oryzacystatin-III numbering), and this region contained a glycine residue (Gly(14)), which is evolutionarily conserved in the cystatin superfamily. We named this novel protein oryzacystatin-III. Nucleotide sequencing of the 5'-flanking region of the oryzacystatin-III gene showed that it is highly homologous to the oryzacystatin-II gene but distinct from the oryzacystatin-II locus. Oryzacystatin-III inhibited papain, ficin, and human cathepsin B. The inhibition constants for papain and ficin differ from those of oryzacystatin-I and -II, and cathepsin B activity is affected only by oryzacystatin-III, showing differences in the interaction of these inhibitors with enzymes. These data suggest that the above three inhibitors may play unique physiological roles in the regulations of rice cysteine proteinases.     

Between and within cultivar variability in salt tolerance in lucerne, (Medicago sativa L.)

Summary The response of two-week-old seedlings of 35 cultivars of lucerne (Medicago sativa L.) to increasing NaCl concentrations was assessed in solution culture under growth room conditions. Shoot length, used as the criterion for assessing salt tolerance, was found to decrease significantly with increasing NaCl concentration for all cultivars, but there was considerable variation in response between, and within cultivars. It is concluded that selection between and within cultivars should lead to increased salt tolerance in this species.     

包膜复合肥对水稻生长及营养特性的影响

试验以早稻品种早凡5号和晚稻品种新香优80为材料,利用自行研制的低成本包膜复合肥(N:P2O5:K2O=17％:7％:12％)设置不同用量处理,以相同养分含量的未包膜复合肥为对照,在大田条件下采用随机区组小区试验,比较研究了供试水稻品种的生长生理和营养生理特性。结果表明包膜肥各处理分蘖增长平缓,成穗率高,而对照前期分蘗发生快,无效分蘗增多。包膜肥各处理剑叶叶绿素含量和SPAD测定值在后期显著高于对照。与等量或减量20％-25％的未包膜复合肥处理比较,早稻产量增加5.0％-12.8％,晚稻实际产量提高7.5％-13.6％,均达5％显著水平。因此,施用包膜复合肥前期养分释放平稳,水稻生长健壮;中后期可提供较多的养分,延长水稻叶片的光合功能期,促进光合产物向籽粒中转运,增加干物质产量和稻谷产量。     

Volatile components in stored rice [Oryza sativa (L.)] of varieties with and without lipoxygenase-3 in seeds.

Lipoxygenase (LOX) is thought to play an important role in the formation of desirable or undesirable flavor and aroma in many plant products. In rice seeds, LOX activity is localized in the bran fraction and LOX-3 is the major isozyme component. We used gas chromatography-mass spectrometry to determine whether the degree of staleness in the flavor of stored brown rice was related to the presence of LOX-3. We found that the amount of hexanal, pentanal, and pentanol in normal raw LOX-3 rice markedly increased during storage at 35 degrees C. That in LOX-3-less rice increased slightly but was a third to a fifth that of normal LOX-3 rice. In cooked rice, the amount of these components from glutinous rice exceeded that in nonglutinous rice, and that in normal LOX-3 rice exceeded that in LOX-3-less rice. These results indicate that the stale flavor production in LOX-3-less rice during storage is less than that in normal LOX-3 rice.     

根癌农杆菌介导的转录因子DREB1A基因在地被菊花中的遗传转化

以地被菊花（Dendranthema grandiflomm cv．White Snow）无菌苗叶片为外植体，在附加不同浓度植物生长调节剂的MS基础培养基上诱导培养，通过器官直接发生途径获得再生植株。结果表明，2．0mg/L 6-BA和0．2mg/L NAA组合可获得93．8％的再生芽。将含拟南芥（Arabidopsis thaliana）逆境诱导转录因子DREB1A基因的植物表达载体pBI-DREB1A导入根癌农杆菌（Agrobacterium tumefaciens）LBA4404，农杆菌介导法遗传转化地被菊花White Snow叶盘，经PCR和PCR—Southem检测，DREB1A基因已整合到转化植株的基因组中。遗传转化过程中，预培养2d后，将OD600=0．5～0．7的根癌农杆菌稀释30倍后侵染叶盘10min，再共培养2d，有利于获得最高遗传转化效率。     

Regulation of anthocyanin accumulation in rice (Oryza sativa L. subsp. indica) using MgSO4 spraying and low temperature

Pericarp pigmented grains of rice have been reported as anthocyanin enriched source of antioxidants. The aim of this investigation was to regulate anthocyanin accumulation in pericarp pigmented rice cv. ‘Hom Nil’ using 0, 10, 50, and 100 mM MgSO₄ foliar spray. The level of magnesium content in the leaf tissues treated with 100 mM MgSO₄ for 2 d increased 2.6 times over control (0 mM MgSO₄). After growing the treated plants under 16°C for 28 d, photosynthetic abilities were reduced by 17?93%, leading to retarded plant height (39?42% less than control). In addition, grain yield per panicle of the plants grown under 16°C for 28 d was reduced by 26?75% compared to the plants grown under 32°C.Total anthocyanin concentration, cyanidin-3-glucoside and peonidin-3-glucoside in the pericarp of the grains derived from the rice treated with 100 mM MgSO₄ was increased 3.8 times over the control. Peonidin-3-glucoside, a dominant compound was reported for the first time. The study concludes that Mg?enrichment may play a key role as a metalloid in stabilizing anthocyanin, a secondary metabolite in the leaf sheath tissues and pericarp of grains in ‘Hom Nil’ rice cultivar.     

籼稻体细胞无性系雄性不育突变体后代遗传及其恢保关系鉴定

对野败型保持系珍汕97B、恢复系IR24、IR26和泰引1号等4个水稻材料的幼穗在不同的培养基上培养、再生植株及对其后代进行育性和测交鉴定及其它遗传分析,探讨了雄性不育变异株的恢保关系,结果表明:在IR26、泰引1号和珍汕97B的R1、R2群体中都获得了不同比率的雄性不育变异株,这些不育株的花粉败育可分为无花粉、典败、圆败和染败4种类型.对离体培养产生的雄性不育变异株用一批现有CMS不育系的保持系和恢复系进行杂交,其中有2株分别来自IR26(编号为269157)和珍汕97B(编号为B29154)的不育株,其恢保关系与野败型一致,现已回交5代.来自IR26(编号为2692411)的另一雄性不育株,用上述亲本已连续回交5代,后代均不育,其恢保关系似与野败不同.而有些来自珍汕97B和IR26的12株雄性不育变异株,杂交后代(F1)全部可育,结实率85%以上.     

大气CO2倍增对水稻生长和发育的影响

矮香糯水稻(Oryza sativa L, )插身后生长在大气（350ppm CO₂）和CO₂倍增（700 ppm CO₂）的开顶式培养室中，结果显示，在CO₂倍增的条件下，矮香糯生长旺盛，根系发达，根系干重增加23%，株高增加12%，每穗结实率增加29%，每株籽粒干重增加41%。本文对目前有关这方面的研究现状进行了讨论。     

Fine root growth and water use efficiency in alfalfa (Medicago sativa L. cv. Gongong No. 1) planted along a salinity gradient in coastal area of Dalian,Northeast China

The response of plant growth to a geographical salinity gradient under the current scenario from inland to seaside implies the possible future trend of plant development in response to sea-level rise driven by global warming. In the present study, alfalfa (Medicago sativa L. cv. Gongnong No. 1) was planted in sites along a salinity gradient in Dalian City, Northeast China. Fine root growth was investigated at three depths of 0–10 cm, 10–20 cm and 20–30 cm. Meanwhile, another control experiment was conducted using potted alfalfa exposed to gradual levels of salinity at the same levels as the salinity gradients in the field to confirm the unique impact of soil salinity on alfalfa physiology. Results from both field and pot experiments indicated that the parameters of net photosynthesis rate (P_N), foliar transpiration (T_r) and water use efficiency (WUE) all decreased with the increase of gradient from inland to seaside. Root surface area also showed a general decline for all three soil depths, but differences among the three soil depths were not significant. Root diameter was greatest in soils at a depth of 0–10 cm, while root tip number was greatest in soils at a depth of 10–20 cm. All root morphological parameters decreased with salinity level along the geographical gradient from inland to coastal seaside, while both root surface area and root tip number positively correlated with soil ammonium and nitrate contents, respectively. Our results predicted that, driven by seawater intrusion from sea-level rise under global warming, future performance of inland alfalfa may suffer from severe soil salinity through constricting new root egress and elongation. This trend would be mainly driven by soil salinity which reduces photosynthesis by stimulating water loss in alfalfa.     

供磷水平对铁、锰、铜、锌在稻株中分布和糙米品质的影响

用室内盆栽试验,研究了不同磷肥施用量对铁、锰、铜、锌在水稻中分布和糙米品质的影响。结果表明,盆栽条件下,施用磷肥显著降低了Fe、Mn、Cu、Zn在IR68144地上部的积累量,显著增加了Fe、Mn、Cu、Zn在两个品种糙米中的分布。在不同磷肥施用水平下,Fe、Mn、Cu、Zn在IR68144地上部的积累量显著高于IR64,表明IR68144从根到地上部运输微量元素Fe、Mn、Cu、Zn的能力更强。此外,施用磷肥促进了富铁水稻糙米对蛋白质的积累,抑制其对直链淀粉的积累,且导致胶稠度变软,对IR64糙米品质的影响较小。说明合理的磷肥管理措施可在一定程度上调控子粒微量元素积累,改善稻米营养品质。     

Black rice (Oryza sativa L. indica) pigmented fraction suppresses both reactive oxygen species and nitric oxide in chemical and biological model systems

Anthocyanins, present in fruits and vegetables as natural colorants, have been well characterized to possess bioactive properties. Anthocyanin components extracted from black rice (Oryza sativa L. indica) separated by gel filtration and identified using LC-MS were cyanidin 3-glucoside and peonidin 3-glucoside. A standardized extract of black rice pigmented fraction (BRE) containing known proportions of cyanidin 3-glucoside and peonidin 3-glucoside exhibited marked antioxidant activities and free radical scavenging capacities in a battery of in vitro model systems. Significant (p < 0.05) prevention of supercoiled DNA strand scission induced by reactive oxygen species (specifically, peroxyl radical and hydroxyl radicals) and suppression of the oxidative modification of human low-density lipoprotein was obtained with BRE. In addition, BRE reduced (p < 0.05) the formation of nitric oxide by suppressing inducible nitric oxide synthase expression in murine macrophage RAW264.7 cells, without introducing cell toxicity. The results of this study show that black rice contains anthocyanin pigments with notable antioxidant and anti-inflammatory properties for potential use in nutraceutical or functional food formulations.     

Biosynthetic mechanism of 2-acetyl-1-pyrroline and its relationship with Delta1-pyrroline-5-carboxylic acid and methylglyoxal in aromatic rice (Oryza sativa L.) callus

2-Acetyl-1-pyrroline (2-AP) was identified as the major flavor compound in aromatic rice varieties Tainung 71 and 72. In order to understand the mechanism of 2-AP biosynthesis in aromatic rice, we studied the formation of putative precursors, Delta(1)-pyrroline-5-carboxylic acid and methylglyoxal. The endogenous Delta(1)-pyrroline-5-carboxylic acid contents of Tainung 71 and 72 calli reached 191 to 276%, compared to nonaromatic rice Tainung 67. In addition, calli of Tainung 71 and 72 contained 1.30- and 1.36-fold, respectively, higher methylglyoxal levels than that of Tainung 67. Specific enzyme activities of Delta(1)-pyrroline-5-carboxylic acid-synthetic enzyme including Delta(1)-pyrolline-5-carboxylic acid synthetase (P5CS) and ornithine aminotransferase (OAT) increased significantly in aromatic rice varieties. The expression levels of P5CS1 and P5CS2 genes were found to be significantly higher in aromatic rice than nonaromatic rice. Results of a tracer experiment with (15)N-labeled glutamic acid revealed that the nitrogen atom of 2-acetyl-1-pyrroline was derived from glutamic acid. Upregulation of P5CS in aromatic rice Tainung 72 may contribute to the increase of Delta(1)-pyrroline-5-carboxylic acid level and thus leads to the accumulation of an extra amount of 2-acetyl-1-pyrroline.     

The role of potassium solubilizing bacteria (KSB) inoculations on grain yield,dry matter remobilization and translocation in rice (Oryza sativa L.)

This experiment was carried out in both pot and field conditions to assess the effects of three native potassium solubilizing bacteria (KSB: including Pantoea agglomerans, Rahnella aquatilis and Pseudomonas orientalis which were isolated from paddy fields) on grain yield (GY), dry matter remobilization (DMR) and translocation (DMT), dry matter translocation efficiency (DME) and contribution of pre-anthesis assimilates (CA) from the leaves and leaves plus stem to the grain in rice (Oryza sativa L). The results indicated that the KSBs significantly increased GY around 20–38% in the pot and 20–52% in the field, especially when half of the recommended potassium fertilizer was applied (K₂So₄, 44% K₂O) as compared to the control. Results also showed that the KSB inoculations and chemical fertilizers had a significant effect on DMR, DMT, DME and CA. In conclusion, these KSBs can be utilized as bio-inoculants for half reduction of K chemical fertilizer consumption in rice production systems.