Changes in soil phosphorus fractions in a calcareous paddy soil under intensive rice cropping

We conducted a 4-year field experiment on a calcareous paddy soil in Zhejiang province of China to measure the changes in chemically extracted soil P fractions in an irrigated double-cropping rice system. Treatments included four fertilizer combinations (unfertilized control, NK, NP, and NPK) as main-plots and two rice cultivar types (inbred vs. hybrid rice) as sub-plots. Total plant P uptake and grain yield of rice declined in all treatments over time. Severe P-deficiency and significant rice yield losses began in treatments without P application after the second rice crop. Compared to inbred rice, hybrid rice increased grain yield (+18%), N uptake (+11%) and K uptake (+27%) but there was no significant difference in total plant P uptake. Recovery efficiencies of fertilizer-P averaged 31–32% in both cultivars. In treatments without P application, the P mass balance was negative (−6 to −8 kg P ha⁻¹ crop⁻¹) and phosphorus was drawn down in all inorganic P fractions, including resin, alkali- (NaHCO₃-Pi and NaOH-Pi) and acid-soluble P fractions (dilute HCl-P, concentrated HCl-P, residual-P). Only small amounts were removed from organic P fractions, 1–3 mg P kg⁻¹ year⁻¹ from NaHCO₃-Po and none from NaOH-Po. In treatments with fertilizer-P addition, the P mass balance was positive (+8 to 10 kg P ha⁻¹ crop⁻¹), soil P declined at a slower rate in inorganic P fractions and it increased (+51%) in the residual-P fraction. Hybrid rice generally caused greater depletion of inorganic soil P fractions than inbred rice, but there was no difference among cultivars in their effect on NaHCO₃-Po and NaOH-Po. Positive correlations (r = 0.63–0.81, P < 0.001) were observed between all inorganic P fractions (except residual-P) and total P uptake by rice. Our results suggest that rice plants draw P from a continuum of chemically extracted fractions that are assumed to have widely differing plant P availability. Regular P additions are required to maintain the effective soil P supply and differences between inbred and hybrid rice should be taken into account in P management strategies.     

Structural and enzymatic characterization of the isoamylase1 homo-oligomer and the isoamylase1–isoamylase2 hetero-oligomer from rice endosperm

The present study established that there are two distinct polymeric forms of isoamylase1 (ISA1) in rice endosperm: presumably a homo-pentamer of ISA1 and a hetero-hexamer composed of five ISA1 and one ISA2. The molecular sizes of the homo- and hetero-oligomers, which could be fractionated by hydrophobic chromatography, were approximately 420–480 and 510–550 kDa, respectively. The hetero-oligomer exhibited higher affinities for various branched polyglucans, especially for phytoglycogen, which had a K _m value that was approximately 12 times lower relative to that with the homo-oligomer, although no marked differences were found in chain preferences for debranching of amylopectin and phytoglycogen between these forms. The hetero-oligomer was active even when incubated at 50°C for 10 min, while the homo-multimer was completely inactivated at 40°C in 10 min. When the ISA1 homo-oligomer was incubated with the ISA2 protein expressed in Escherichia coli and applied onto a nondenature polyacrylamide gel, additional debranching activity bands which were specific for the purified ISA1–ISA2 preparation were also detected, indicating that ISA1 and ISA2 combine to form a hetero-oligomer. These results suggest that the hetero-oligomer plays a predominant role in the amylopectin biosynthesis in rice endosperm although the homo-oligomer can complement the function of the hetero-oligomer at least to some extent.Electronic Supplementary Material Supplementary material is available in the online version of this article at and is accessible for authorized users.     

水稻淀粉合成的分子生物学研究进展

本文综述了水稻淀粉合成的分子生物学研究的最新进展,主要内容是参与淀粉合成的酶鉴定及其基因表达调控,也介绍了对这些酶的遗传操作改良稻米淀粉品质等内容     

Temperature induced changes in the starch components and biosynthetic enzymes of two rice varieties

The effects of temperature on starch and amylose accumulation, fine structure of amylopectin and activities of some enzymes related to starch synthesis in developing rice endosperms was examined. Two early indica rice varieties were used, differing in amylose concentration (AC, %), namely Jia 935 (low AC) and Jia 353 (high AC). The results showed that the effects of high temperature on AC and amylopectin fine structure were variety-dependent. High temperature caused a reduction in amylose concentration and an increase in the short chain (CL<22) proportion of amylopectin for Jia 935; while opposite was true for Jia 353. High temperature also reduced and increased the activity of granule-bound starch synthase (GBSS) in Jia 935 and in Jia 353, respectively. This suggests that a change in the ratio of amylose/starch due to temperature was attributable to a change in GBSS activity. Moreover, obvious differences between the two rice varieties were detected in the activities of sucrose synthase (SuSy), ADP-glucose pyrophosphorylase (ADPG-Ppase), soluble starch synthase (SSS), starch branching enzyme (SBE), starch de-branching enzyme (SDBE) and starch phosphorylase (SPase) to high temperature. Accumulation rate of amylose was significantly and positively correlated with GBSS for Jia 935, but not for Jia 353. Amylose accumulation was also significantly and positively correlated with the activities of SDBE, SBE, ADPG-Ppase and SuSy for both varieties. The results suggest that the ratio of amylose to starch in rice endosperm is not only related to GBSS, but also affected by the activities of SDBE, SBE, ADPG-Ppase and SuSy.     

Pyramiding transgenes for multiple resistance in rice against bacterial blight,yellow stem borer and sheath blight

The present investigation revealed that the alk and gel(t) genes, which cause the differences between a japonica rice variety Nipponbare and an indica rice variety Kasalath in terms of the disintegration of endosperm starch granules in alkali solution and their gelatinisation in a 4 M urea solution, respectively, cosegregated in backcross inbred lines derived from a cross between the two varieties. The segregation pattern of the profile for amylopectin chain-length, which was distinguished by enrichment in short chains of DP≦11 and depletion in intermediate-size chains of 12≦DP≦24 in japonica as compared with indica, was exactly the same as those of the above physico-chemical properties of starch granules, and the gene was designated as acl(t). Gene-mapping analysis showed that the starch synthase IIa (SSIIa) gene is located at the alk locus on chromosome 6 in the rice genome. These results lead us to the possibility that different alleles of the SSIIa gene are responsible for differences in amylopectin structure between the two varieties, in that SSIIa plays a distinct role in the elongation of short chains within clusters (A+B₁ chains) of amylopectin. It is proposed that the activity of SSIIa in japonica rice is reduced in amount or functional capacity relative to the activity of this enzyme in indica rice. This, in turn, would explain why starch from japonica rice has a lower gelatinisation temperature than starch from indica rice and is more susceptible to disintegration in alkali or urea. The evidence for this hypothesis is that the alk(t), gel(t), acl(t) and SSIIa genes all map to the same locus. Received: 29 January 2001 / Accepted: 12 April 2001     

Particle size distribution of rice flour affecting the starch enzymatic hydrolysis and hydration properties

Rice flour is becoming very attractive as raw material, but there is lack of information about the influence of particle size on its functional properties and starch digestibility. This study evaluates the degree of dependence of the rice flour functional properties, mainly derived from starch behavior, with the particle size distribution. Hydration properties of flours and gels and starch enzymatic hydrolysis of individual fractions were assessed. Particle size heterogeneity on rice flour significantly affected functional properties and starch features, at room temperature and also after gelatinization; and the extent of that effect was grain type dependent. Particle size heterogeneity on rice flour induces different pattern in starch enzymatic hydrolysis, with the long grain having slower hydrolysis as indicated the rate constant (k). No correlation between starch digestibility and hydration properties or the protein content was observed. It seems that in intact granules interactions with other grain components must be taken into account. Overall, particle size fractionation of rice flour might be advisable for selecting specific physico-chemical properties.     

云南30个水稻品种对水稻白叶枯病的抗性评价

【目的】水稻白叶枯病是一种严重危害水稻的细菌性病害,培育抗性品种是治理该病害的重要途径。因此,明确云南水稻材料对检疫性病害水稻白叶枯病的抗性,可以为该病害的防治与监测、水稻栽培的合理布局和良好抗性资源的获取提供依据。【方法】采用剪叶接种法测定云南稻区30个品种对7个不同致病型白叶枯病菌的抗性。【结果】在供试的30个云南水稻品种中,2个品种(玉粳16和JS42糯稻)对7个不同致病型菌株均表现为抗性;15个品种对7个致病型菌株均表现感病;对HEN11、SCYC-6、YN7、YN11、FUJ、YN241和PX099等7个致病型菌株表现抗性的水稻品种分别占26.67%、16.67%、23.33%、13.33%、6.67%、10.00%和20.00%。此外,区试材料的抗性比例高于主栽品种,地方稻未发现抗性品种。【结论】现在生产上的大部分水稻品种对优势致病型病原菌入侵的抵抗能力降低甚至丧失。针对云南地区的优势致病小种FUJ筛选得到2个抗性品种:玉粳16和JS42糯稻。     

The starch granule associated proteomes of commercially purified starch reference materials from rice and maize

Commercially available reference materials are integral components of many experimental protocols, as it is critical to compare one's results to those derived from well-characterized standards. Most reference materials are well defined, with all their components being cataloged. However, certain reference materials, such as commercially prepared starch samples, can have undefined components, potentially limiting their usefulness as standards. The proteome of commercially prepared starch has not been documented, and to that end, we initiated a mass spectrometry-based survey of the proteins associated with starch granules in commercially prepared rice and maize starch samples. We performed direct trypsin treatments of starch samples and sequenced both the water-soluble peptides liberated into the aqueous supernatant and the peptides released from the starch granule surface by isopropanol solvent washing. We discovered that the majority of proteins, in both rice and maize samples, were involved in either carbohydrate metabolism or storage. We also documented proteins that are markers for seed maturity and for starch mobilization.     

Rice endosperm-specific plastidial α-glucan phosphorylase is important for synthesis of short-chain malto-oligosaccharides

Previous genetic studies have indicated that the type L α-glucan phosphorylase (Pho1) has an essential role during the initiation process of starch biosynthesis during rice seed development. To gain insight into its role in starch metabolism, we characterized the enzymatic properties of the Pho1 recombinant form. Pho1 has significantly higher catalytic efficiency toward both linear and branched α-glucans in the synthesis direction than in the degradation direction with equilibrium constants for the various substrates ranging from 13 to 45. Pho1 activity is strongly inhibited by its own reaction product (Pi) in the synthesis reaction (K_i = 0.69 mM) when amylopectin is the primer substrate, but this inhibition is less pronounced (K_i = 14.2 mM) when short α-glucan chains are used as primers. Interestingly, even in the presence of Pi alone, Pho1 not only degrades maltohexaose but also extends them to synthesize longer MOSs. Production of a broad spectrum of MOSs (G4-G19) was stimulated by both Pi and Glc1P in an additive fashion. Thus, even under physiological conditions of high Pi/Glc1P, Pho1 extends the chain length of short MOSs which can then be used as subsequent primer by starch synthase activities. As ADP-glucose strongly inhibits Pho1activity, Pho1 likely operates only during the initial stage and not during maturation phase of starch synthesis.     

Expression of a fungal glucoamylase in transgenic rice seeds

Glucoamylase, which catalyses the hydrolysis of the α-1,4 glycosidic bonds of starch, is an important industrial enzyme used in starch enzymatic saccharification. In this study, a glucoamylase gene from Aspergillus awamori, under the control of the promoter of seed storage protein Gt1, was introduced into rice by Agrobacterium-mediated transformation. Significant glucoamylase activity was detected specifically in the seeds but not other tissues of the transgenic rice lines. The highest enzymatic activity was found in the transgenic line Bg17-2, which was estimated to have about 500 units per gram of seeds (one unit is defined as the amount of enzyme that produces 1 μmol of reducing sugar in 1 min at 60 °C using soluble starch as substrate). The optimum pH for the activity of the rice produced enzyme is 5.0–5.5, and the optimum temperature is around 60 °C. One part of this transgenic glucoamylase rice seed flour fully converted 25 parts of corn starch pre-liquefied by an α-amylase also produced by a transgenic rice into glucose in 16 h incubation. This study suggests that this hydrolysis enzyme may substitute commercial fermentation enzymes for industrial starch conversion.     

Architectural changes of heated mungbean, rice and cassava starch granules: Effects of hydrocolloids and protein-containing envelope

Architectural changes of starch granules induced by heat were demonstrated using light microscopy and confocal laser scanning microscopy. Heat treatment (80 °C, 30 min) on mungbean starch, cassava starch and rice flour suspensions resulted in the rearrangement of amylose and granule-associated proteins within the deformed granules. The presence of alginate and carrageenan influenced the RVA pasting characteristics of starch/flour-hydrocolloid mixed suspensions by maintaining the granular structure of amylose-rich swollen granules or inducing the aggregation of the swollen ones. Generally, the addition of hydrocolloid increased peak viscosity, lowered breakdown and reduced setback of the flour-hydrocolloid mixed paste. This study demonstrated that the heat treatment in excess water generated the protein-containing granule envelope encasing the mungbean and cassava starch content within the deformed granules.     

Association of waxy gene single nucleotide polymorphisms with starch characteristics in rice (Oryza sativa L.)

The waxy gene, which encodes the granule bound starch synthase enzyme, is one of the key genes influencing starch synthesis in the rice endosperm. To investigate functional differences between GBSS alleles, we cloned and sequenced GBSS cDNA from a series of cultivars that differed substantially in apparent amylose content and starch viscosity characteristics. We found two single nucleotide polymorphisms in exons 6 and 10 that resulted in amino acid substitutions. These substitutions are associated with differences in apparent amylose content and viscosity characteristics. Subsequent sequencing of these regions from additional cultivars confirmed their association with particular rice quality characteristics. These point mutations could prove useful as molecular markers in the production of cultivars with superior eating, cooking and processing quality, and contribute to our understanding of the various structural and functional differences among granule bound starch synthase alleles.     

Use of Three Different Marker Systems to Estimate Genetic Diversity of Indian Elite Rice Varieties

Genetic diversity among 42 Indian elite rice varieties, which is important for selection of parents for conventional breeding and hybrid program, was evaluated using three different types of DNA markers and parentage analysis. Random amplified polymorphic DNA (RAPD), inter-simple sequence repeat (ISSR) and sequence tagged microsatellite site (STMS) markers resulted in mean heterozygosity values of 0.429, 0.675 and 0.882 over all loci, respectively, and marker index values of 2.21, 4.05 and 5.49, respectively. The three molecular marker systems together provide wider genome coverage and, therefore, would be a better indicator of the genetic relationships among the 42 elite rice cultivars than those revealed using individual molecular markers. A total of 153 bands (91%) were polymorphic out of 168 bands amplified, considering all the markers together. The average genetic similarity coefficient across all the 861 cultivar pairs was 0.70 while the average coefficient of parentage was 0.10. Cluster analysis revealed that there was a very poor correlation (correlation coefficient <0.1) between dendrograms generated using coefficients of parentage and molecular marker generated genetic similarities, which can be attributed to selection pressure, genetic drift, sampling of loci and unknown relationships among supposedly unrelated ancestors. This revised version was published online in July 2006 with corrections to the Cover Date.     

淹涝胁迫对水稻叶鞘和叶片中淀粉粒分布的影响

淹涝是世界上当前所面临的最严重自然灾害之一,尤其是近几年来,淹涝给我国的农业生产带来了巨大损失,全国平均每年受涝面积814万hm2,其中成灾面积448万hm2,损失粮食约28亿kg〔1〕。水稻是我国的主要粮食作物,并且分布于淹涝易发生的南方多雨潮湿地区和北方低洼地带,淹涝对水稻产量影响极大,因此,研究淹涝胁迫对水稻的伤害及水稻的耐淹机制越来越受到人们的重视〔2〕。淹涝对植物的伤害并非是因为水分过多而造成的直接伤害,而是由于淹涝造成的次生胁迫,其中最严重的是缺氧。在缺氧条件下,细胞中的碳水化合物代谢途径改变,对植物的生长和发育…     

Distribution of zinc fractions and their transformation in submerged rice soils

Distribution of different forms of Zn in 16 acid alluvial rice growing soils of West Bengal (India) and their transformation on submergence were studied. The results showed that more than 84% of total Zn occurred in the relatively inactive clay lattice-bound form while a smaller fractionviz. 1.1, 1.6, 11.1 and 2.0 per cent of the total occurred as water-soluble plus exchangeable, organic complexed, amorphous sesquioxide-bound and crystalline sesquioxide bound forms, respectively. All these four Zn forms showed significant negative correlations with soil pH (r=−0.48^**, −0.39^*, −0.61^** and −0.67^**, respectively), while the latter two Zn forms showed significant positive correlations with Fe₂O₃ (0.68^** and 0.88^***) and Al₂O₃ (0.89^*** and 0.75^***) content of the soils. The different Zn forms were found to have positive and significant correlations amongst each other, suggesting the existence of a dynamic equilibrium of these forms in soil. Submergence caused an increase in the amorphous sesquioxide-bound form of Zn and a decrease in each of the other three forms. The magnitude of such decreases in water-soluble plus exchangeable and crystalline sesquioxide-bound forms was found to be correlated negatively with initial pH values of the soils and positively with the increase in the amorphous sesquioxide-bound form, indicating their adsorption on the surface of the freshly formed hydrated oxides of Fe, which view was supported by the existence of significant positive correlation between the increase in the amorphous sesquioxide-bound form of Zn and that in AlCl₃-extractable iron. The existence of a positive correlation between the decrease in crystalline sesquioxide-bound Zn and that in Fe₂O₃ content in soil suggested that on waterlogging the soil Zn occluded in the cry talline sesquioxide was released as a result of reduction of Fe₂O₃.     

粳稻支链淀粉合成相关基因的遗传组成及其对品质的影响

摘要：淀粉的组成和结构是决定稻米品质的主要因素。利用17个淀粉合成相关基因位点的分子标记鉴定67个粳稻品种的淀粉合成相关基因的籼粳来源,检测表明67个品种均带粳型Wx基因,GBSSⅡ、SSⅡ-2和SSⅣ-1位点没有发生重组,其他13个支链淀粉合成相关基因位点有籼型等位基因的渗入,渗入频率在1.9%-46.27%之间,其中SSⅠ籼型等位基因渗入到粳稻品种中最高;逐步回归分析表明有7个基因对10个淀粉理化指标有效应,其中Isa对最高粘度、崩解值、回复值、消减值和峰值时间有效应,其它基因只对1-4个性状有明显的遗传效应。说明在支链淀粉合成相关基因座上籼粳等位基因已经发生了明显的分化,具有不同的遗传功能,为水稻优质品种培育提供了参考依据。     

Field performance and cytology of protoplast-derived rice (Oryza sativa): high yield and low degree of variation of four japonica cultivars

Summary Protoplast-derived rice plants of four Japanese cultivars, Nipponbare, Fujisaka 5, Norin 14 and Iwaimochi were individually cultivated in a submerged paddy field. They exhibited more stems, which resulted in more panicles than respective control plants. Other characteristics of protoplast-derived plants were (compared with controls): a slightly shorter or similar culm length, fewer spikelets per panicle, slightly lower seed fertility and similar or lighter 1,000 kernel weight. Grain yield of protoplast-derived plants was more than that of respective control plants in four cultivars. The cause of the higher yield of protoplast-derived plants seems to be mainly due to increased panicle number. Among 126 protoplast-derived plants, 1 triploid, 10 tetraploids and 1 aneuploid were found. Furthermore, 11 variants with low seed fertility showing no gross chromosomal anomalies and one plant with abnormal panicles were found. In total, about 80% of protoplastderived plants showed normal characters. The present results are encouraging for the possibility of rice breeding using protoplasts.This paper is dedicated to Professor Emeritus Dr. Shigeyasu Akai on the occasion of his 77th birthday (Kijyu: one of the celebrated ages in Japan)