水稻淀粉合成对夜温变化的响应

目的 探究夜温变化对水稻淀粉形成的影响及其生理机制【方法】以优质软米浙禾香2号为材料,在灌浆初期设置31℃/20℃ (LT)、31℃/24℃ (NT)、31℃/28℃ (HT)3个夜间温度模式,测定其直链淀粉和支链淀粉含量及合成关键酶活性及相关基因的表达。结果 1)与NT相比,LT和HT处理显著降低粒重和淀粉积累,降低糊化温度和胶稠度,并影响支链淀粉链长,降低支链淀粉含量,提高直链淀粉含量,HT的影响要大于LT;2) LT和HT处理对白天叶片净光合速率的影响不显著,但显著降低籽粒中非结构性碳水化合物积累,抑制蔗糖转运基因OsSUT1、OsSUT2和OsSUT4在夜间和白天表达;3) LT 和HT处理降低夜间和白天蔗糖水解相关酶活性,增加淀粉水解酶活性,导致可溶性糖含量升高,籽粒中糖利用受阻;4)与NT相比,LT和HT处理下腺苷二磷酸葡萄糖含量呈现白天降低而夜晚升高的趋势,腺苷二磷酸葡萄糖积累及利用受到抑制,颗粒结合淀粉合酶活性随处理时间延长而显著降低,且白天酶活性也受夜间温度的影响;5)与NT相比,LT和HT处理降低了夜间支链淀粉合成相关酶活性,抑制了夜间相关基因的表达,导致支链淀粉合成受阻,但对白天酶活及相关基因表达的影响不大。结论 夜间高温对淀粉积累的影响要大于夜温降低,夜间高温/低温抑制全天蔗糖转运及代谢,进而抑制淀粉积累;支链淀粉合成受阻是导致直链淀粉相对含量升高的主要原因,直链淀粉合成相关酶活性(白天)受夜温变化影响,而支链淀粉合成相关酶活性(白天)受夜温变化的影响不显著。     

花后水稻穗部夜间远红外增温处理对稻米品质的影响

 以武育粳3号为材料,在开放环境下使用远红外加热器对水稻穗部进行增温处理,研究了灌浆结实期穗部日间或夜间增温对稻米品质的影响。结果表明,夜间增温降低了糙米率、精米率和整精米率,增加了垩白发生,降低了稻米的加工品质和外观品质,与日间增温影响一致。夜间增温提高了稻米蛋白质含量,与日间增温相似,但夜间增温有降低稻米直链淀粉含量的趋势,与日间增温相反。夜间增温处理淀粉结构与日间增温差异较分明,尤其是胚乳腹部,夜间增温胚乳腹部的淀粉粒个体较大,棱角分明,排列紧密;而日间增温淀粉体较小,呈圆形和椭圆形,但数量较多。夜间增温对淀粉相对结晶度的影响与日间增温相似,但夜间增温增幅较小。总体来说,灌浆结实期夜间增温对稻米的加工品质、外观品质、营养品质影响与日间增温一致,对蒸煮食味品质和淀粉结构的影响与日间增温存在很大的差异,且夜间增温对稻米品质的影响程度整体上小于日间增温。     

Effect of Temperature at Grain Filling Stage on Activities of Key Enzymes Related to Starch Synthesis and Grain Quality of Rice

Three japonica rice varieties with different cooking and eating quality were grown at high temperature in the greenhouse and natural field. Effects of temperature at the grain filling stage on these varieties were investigated in terms of the activities of key enzymes related to starch synthesis and cooking and eating quality of rice grain. The high temperature at the grain filling stage increased protein content, and decreased amylose content and taste meter value of rice; inferior grain quality varieties showed a greater magnitude of the increase or decrease than the superior ones. Reaction of rapid visco analyser profiles to the temperature varied with rice varieties. The activities of adenosine diphosphoglucose pyrophosphoryiase （AGPP）, soluble starch synthase （SSS） and starch branching enzyme （SBE） gradually increased to a peak value, and thereafter declined as grain filling progressed. Enzyme activities in different varieties differed in a same filling stage, and also in the time when the enzyme activity reached a maximum. AGPP and SSS were insensitive to the environmental temperature, but SBE was comparatively sensitive to the temperature, and its activity declined when temperature was too high or too low.     

灌浆期夜间高温胁迫下耐热和热敏感水稻籽粒的比较蛋白质组分析

【目的】为了阐明水稻灌浆期耐夜间高温的分子机制,提高水稻耐热性,鉴定了水稻近等基因系耐热纯系XN0437T与热敏感纯系XN0437S在灌浆期夜间高温胁迫下的差异表达蛋白质。【方法】采用桶栽方法培育水稻,于开花期标记同一天扬花的颖花以保障所取颖花样品的生育进程一致;于水稻灌浆初期(花后第8天)进行夜间高温处理。高温处理结束后剪取带标记的颖花提取水稻籽粒总蛋白质,采用8-plex i TRAQ试剂盒进行蛋白质样品的差异显色标记,标记样品采用高效液相系统进行质谱鉴定及定量分析。【结果】鉴定并定量了3130个蛋白质,耐热与热敏感水稻纯系间存在36个差异表达蛋白质。蛋白质功能注释结果显示,鉴定的36个差异表达蛋白质中仅14个蛋白质(占38.9%)注释了功能,12个蛋白质(占33.3%)为推测性功能注释,10个(占27.8%)为功能未知的蛋白质。已注释功能的14个蛋白质中,5个蛋白质参与能量代谢,3个蛋白质参与物质转运与代谢,2个蛋白质参与光合作用,3个蛋白质为响应逆境的锌指蛋白质,1个蛋白质为响应逆境的热激蛋白质。【结论】灌浆期夜间高温影响水稻籽粒细胞内参与能量代谢、物质转运与代谢、光合作用等相关蛋白质的表达模式。水稻籽粒细胞中锌指蛋白质Q67TK9、Q10N88上调表达,锌指蛋白质Q5YLY5下调表达,有利于提高水稻灌浆期对夜间高温的耐热性。     

高温胁迫下水稻胚乳淀粉分支酶各同工型基因的表达特征

 以稻米品质温度敏感型的早籼稻品种浙辐49为材料,利用人工气候箱控温处理试验和实时荧光定量PCR技术,探讨了水稻发育胚乳中淀粉分支酶(starch branching enzyme, SBE)的3种主要同工型基因 (SBEⅠ、 SBEⅢ和SBEⅣ) 在不同温度处理下的相对表达量差异及动态变化特征。结果表明,高温处理对水稻胚乳中SBE基因的表达调控因其同工型的种类而异,SBEⅣ基因在高温处理下呈上调表达,而SBEⅠ和SBEⅢ基因在高温处理下的相对表达量则明显降低,表现为下调表达;与SBEⅣ等同工型相比,水稻胚乳中SBEⅠ基因对高温胁迫的表达响应相对更敏感。     

灌浆成熟期温度对水稻籽粒淀粉合成关键酶活性及品质的影响

 选用蒸煮食味品质不同的3个粳稻品种，利用温室高温和自然常温，研究了灌浆成熟期温度对水稻籽粒淀粉合成关键酶活性及蒸煮食味品质的影响。结果表明，灌浆成熟期高温条件下，水稻籽粒蛋白质含量提高，直链淀粉含量和味度值降低，劣质品种提高或降低的程度大于优质品种；RVA谱特性对灌浆成熟期温度的反应因品种和特性而异；灌浆过程中，籽粒ADPG焦磷酸化酶、可溶性淀粉合成酶和淀粉分支酶活性随灌浆进程，酶活性逐渐增加，达到峰值以后，酶活性又逐渐下降，呈单峰曲线，但不同品种酶活性达到峰值的时间和同一时期的酶活性大小有差异；ADPG焦磷酸化酶和可溶性淀粉合成酶的活性表现对温度的影响反应较为迟钝，而淀粉分支酶活性表现对温度变化的反应较为敏感，温度过高过低都降低该酶活性。     

水稻胚乳淀粉合成相关酶的结构、功能及其互作研究进展

淀粉作为稻米最主要的储藏物质,其生物合成过程直接影响水稻的产量和品质。水稻淀粉的生物合成在造粉体中通过一系列酶促反应完成。本文综述了淀粉合成过程中ADP-葡萄糖焦磷酸化酶(AGPase)、淀粉合酶(SSs)、淀粉分支酶(BE)及淀粉脱支酶(DBE)的结构、功能以及各酶之间的相互作用的最新研究进展,以期为稻米品质改良提供理论参考依据。     

富含抗性淀粉水稻突变体的淀粉特性

 从杂交水稻优异恢复系R7954诱变筛选了1个富含抗性淀粉的突变体RS111。该突变体热米饭中抗性淀粉的含量高达7.0%，是其野生型的2.4倍。突变体的淀粉颗粒形态、DSC和多晶衍射的曲线和参数与野生型明显不同，表现为淀粉颗粒大小较为一致，圆形和卵形淀粉颗粒所占比率较高，溶晶起始温度(TO)、最高温度(TP)、回落温度(TC)、焓变(ΔHGEL)及结晶度更低。突变体的表观直链淀粉、粗脂肪和粗纤维的含量显著提高。     

齐穗至乳熟期不同温度对水稻南粳9108籽粒激素含量、淀粉积累及其合成关键酶活性的影响

【目的】为了明确灌浆结实前中期不同温度对晚粳稻籽粒灌浆、总淀粉及其组分含量的影响特征及其机理,【方法】以优质食味粳稻南粳9108为材料,利用植物培养箱控温,于水稻灌浆结实期设置高温(HT,日均温度31℃)、中温(MT,日均温度26℃)和低温(LT,日均温度21℃)三个处理,以自然温度为对照(CK),研究灌浆结实前中期持续高低温对晚粳稻籽粒灌浆、内源激素含量、淀粉及其及合成关键酶活性的影响。【结果】随温度上升(21℃～31℃),晚粳稻结实率和千粒重降低;灌浆前中期籽粒总淀粉积累速率加快,但持续时间短,总含量降低,而直链淀粉含量及其所占比例在整个灌浆期均呈降低趋势;灌浆前中期脱落酸(ABA)含量显著增加,赤霉素(GA3)含量降低,玉米素核苷(ZR)含量呈"先增先降"趋势;MT处理有利于提高灌浆结实期籽粒生长素(IAA)含量,HT和LT处理均降低籽粒IAA含量,LT处理下,IAA含量先降后升,而HT处理籽粒IAA含量一直处于较低水平;籽粒可溶性淀粉合成酶(SSS)活性与ZR变化规律相似,也呈"先增先降"趋势,但二者在灌浆结实期并不同步;淀粉分支酶(SBE)活性增加而淀粉脱分支酶(DBE)活性降低。【结论】不同温度显著影响晚粳稻籽粒内源激素含量和淀粉合成关键酶活性的变化,进而影响晚粳稻籽粒灌浆、总淀粉及其组分的积累。     

Effects of Weak Light on Starch Accumulation and Starch Synthesis Enzyme Activities in Rice at the Grain Filling Stage

Photosynthetic product in rice plant is transported to grains mainly in the form of sucrose. Sucrose in the grains eventually becomes starch through a series of enzymatic reactions [1-2]. The starch weight accounts for 90% of the brown rice [3-4], so the course of the rice grain filling is the biochemical process in which starch is mainly formed, and the factor of light is indispensable at the course. Much research has been conducted to study the influence of the light on the grain starch synt…     

水稻糖质胚乳突变体Sug-11籽粒灌浆过程的淀粉合成关键酶活性及其与淀粉理化特性关系

以水稻糖质胚乳突变体Sug-11与其野生型对照中花11为材料,通过对两者籽粒中可溶性总糖、蔗糖含量和淀粉含量以及有关淀粉品质理化指标的比较,结合籽粒灌浆过程中糖类物质含量、淀粉合成代谢关键酶活性和相关同工型基因转录表达水平的动态测定,从籽粒淀粉合成代谢角度,对水稻糖质突变体Sug-11的籽粒糖类含量变化和千粒重下降的生理原因进行了分析。结果表明,Sug-11糖质突变体与其野生型在灌浆初期的可溶性糖和蔗糖含量差异并不明显,随着籽粒灌浆进程,两者间的籽粒糖分含量差异在灌浆中后期逐步趋于明显;与野生型相比,Sug-11糖质胚乳突变体的稻米直链淀粉含量和直链淀粉碘蓝值显著下降,而淀粉溶解度和支链淀粉碘蓝值则显著升高,糖质胚乳突变对稻米淀粉的理化特性也产生了明显的影响;在籽粒淀粉合成代谢的几个关键酶中,Sug-11糖质突变体籽粒中的DBE活性及其在灌浆过程中的动态变化与其野生型存在明显差异,揭示了胚乳糖质突变体Sug-11籽粒中淀粉积累减少、糖分含量增加主要是由籽粒灌浆中后期的PUL转录表达水平和DBE活性的大幅下降所引起的,而Sug-11的籽粒灌浆不良和千粒重下降等现象,则与其ADPGase活性在籽粒灌浆前期的显著下降存在一定的联系。     

Soaking Induced Changes in Chemical Composition,Glycemic Index and Starch Characteristics of Basmati Rice

An attempt was made to determine the qualitative changes in basmati rice(Pusa Basmati 1121, PB1121) during soaking at 40 ℃ to 80 ℃. Soaking temperature had significant effect(α = 0.01) on chemical composition, glycemic index and starch characteristics of rice. Starch content, apparent amylose content, crude protein content and crude fat content in un-soaked rice were found to be 73.24%, 27.26%, 8.79% and 2.56%, respectively, but differences in these traits were observed after soaking. Amylose to amylopectin ratio(Am/Ap) decreased from 0.59 to 0.52(soaked at 80 ℃). Crude fibre and crude ash contents increased after soaking. The mineral composition(K, P, S, Ca, Mg, Mn, Fe, Cu and Zn) in soaked rice was found to be 16.46% higher than un-soaked rice at the same degree of polishing. Glycemic index of un-soaked rice was found to be 58.41, but decreased to 54.31 after soaking at 80 ℃. Pasting properties, scanning electron microscope images, and X-ray diffractograms suggested partial gelatinization of starch in the temperature range of 60 ℃ to 80 ℃. Based on qualitative changes in rice(apparent amylose content, Am/Ap ratio and crystallinity rate), it was concluded that intermediate soaking temperatures(60 ℃ to 70 ℃) would be useful for soaking of PB1121.     

Temperature Stress at Grain Filling Stage Mediates Expression of Three Isoform Genes Encoding Starch Branching Enzymes in Rice Endosperm

An early-maturity indica rice variety Zhefu 49, whose grain quality and starch structure are sensitive to environmental temperature, was subjected to different temperatures (32°C for high temperature and 22°C for optimum temperature) at the grain filling stage in plant growth chambers, and the different expressions of three isoform genes (SBEI, SBEIII and SBEIV) encoding starch branching enzyme (SBE) in the endosperms were studied by the real-time fluorescence quantitative PCR (FQ-PCR) method. Effects of high temperature on the SBE expression in developing rice endosperms were isoform-dependent. High temperature significantly down-regulated the expressions of SBEI and SBEIII, while up-regulated the expression of SBEIV. Compared with SBEIV and SBEIII, the expression of SBEI gene in Zhefu 49 rice endosperms was more sensitive to temperature variation at the grain filling stage. This study indicates that changes in weather/climate conditions especially temperature stress influence rice grain formation and its quality as evidenced by isoform expression.     

水稻糯性突变对淀粉理化特性的影响

比较了籼、粳两个糯性突变品种与原亲本品种淀粉的理化特性。糯性突变后除了直链淀粉含量显著降低外,淀粉的糊化温度变高、胶稠度增大、RVA谱(rapid viscosity analyzer)参数下降。RVA谱参数降低的幅度从高到低依次是消减值>回复值>冷胶粘度>热浆粘度>最高粘度>崩解值,表明控制直链淀粉含量的Wx位点对淀粉糊化特性和粘滞性是有影响的,存在多效性。同时,两个糯稻品种之间的RVA谱差异较大,其中崩解值和最高粘度差值均大于糯性突变对它们的影响,说明除直链淀粉外,支链淀粉也影响淀粉的理化特性。对Wx位点鉴定的结果显示扬辐糯4号wx基因第一内含子剪切位点＋1位的碱基由原品种的G突变成了T,扬糯2号与其原品种相同(均为T);两个糯稻突变品种wx基因前导区的(CT)n微卫星重复序列与原品种相比都发生了变化。     

稻谷碎米淀粉性能改良研究初报

试验采用6种不同处理方法对碎米淀粉性能改良作了初步的探讨和研究,结果表明:碎米放置在温度42℃、湿度88%的老化箱内进行人工加速老化11d后,胶稠度由原来的44.67mm下降为33.33mm,胶稠度显著降低,但该方法是否适用于米粉加工还有待在生产实践中进一步验证。     

Accurate Measurement of Pasting Temperature by the Rapid Visco-Analyser:a Case Study Using Rice Flour

Pasting properties are among the most important characteristics of starch,determining its applications in food processing and other industries.Pasting temperature derived from the Rapid Visco-analyser(RVA)(Newport Scientific),in most cases,is overestimated by the Thermocline for Windows software program.Here,two methods facilitating accurate where the pasting viscosity begins to increase,the other is to manually record the time(T1)when the pasting viscosity begins to increase and calculate the pasting temperature with the formula of(45/3.8)×(T1-1) 50 for rice flour.The latter method gave a manually determined pasting temperature which was significantly correlated with the gelatinization temperature measured by differential scanning calorimetry.     

Relationship Between Degree of Starch Gelatinization and Quality Attributes of Parboiled Rice During Steaming

Paddy rice samples were parboiled by soaking at 65 °C for 180 min and steaming at 96 °C for 2–10 min, and then dried to achieve the final moisture content of 11% ± 1%. The degree of starch gelatinization(DSG) and several quality attributes(head rice yield(HRY), color value and hardness) of parboiled rice were measured. Results showed that DSG(46.8%–77.9%), color value(18.08–19.04) and hardness(118.6–219.2 N) all increased following steaming. In contrast, the HRY increased(64.8%–67.1%) for steaming times between 2–4 min but decreased(67.1%–65.0%) for steaming times between 4–10 min. Linear relations between DSG and color value(R~2 = 0.87), and DSG and hardness(R~2 = 0.88) were observed. The suitable DSG of parboiled rice leading to the highest HRY was found to be 62.5%, obtained following 4 min of steaming.     

高温胁迫下黄嘌呤脱氢酶基因超表达对水稻幼苗的保护作用

【目的】黄嘌呤脱氢酶(xanthine dehydrogenase, XDH)是嘌呤代谢的关键酶。通过分析高温胁迫对OsXDH超表达转基因水稻株系幼苗叶片生理指标的影响,探究XDH缓解水稻高温胁迫的生理机制。【方法】以OsXDH超表达转基因株系(xdh1和xdh5)及受体品种日本晴(WT)为材料,研究了高温胁迫对水稻幼苗叶片叶绿素含量、相对含水量、可溶性蛋白含量、活性氧代谢、抗氧化酶活性、XDH活性及嘌呤代谢产物尿囊素(allantoin)和尿囊酸(allantoate)含量等生理指标的影响。【结果】高温胁迫处理前,超表达株系的叶绿素含量、相对含水量、可溶性蛋白含量、活性氧代谢及抗氧化酶活性等生理指标均与野生型无显著差异;高温胁迫5 d,超表达株系的叶绿素、相对含水量、可溶性蛋白含量及抗氧化酶活性均高于野生型,而过氧化氢(H₂O₂)及丙二醛(MDA)的含量显著低于野生型;适温恢复生长5 d,野生型和超表达株系的叶绿素含量、相对含水量及可溶性蛋白含量均有所提高,超表达株系均高于野生型,H₂O₂ 和MDA的含量降低,超表达株系均低于野生型;受高温诱导超表达株系与野生型中XDH酶活性及尿囊素和尿囊酸的含量均提高,且在整个处理过程中超表达转基因株系均高于野生型。【结论】XDH通过调控酰脲类物质的合成,补偿自身的抗氧化能力并增强抗氧化酶系统的活性,从而有效提高水稻幼苗对高温胁迫的耐受能力。     

应用RVA测定米粉淀粉成糊温度

淀粉的糊化温度是稻米淀粉品质的重要指标,决定淀粉在食品及工业上的用途,也与食用稻米的蒸煮品质密切相关。淀粉的糊化温度可以用淀粉谱仪（如快速淀粉黏滞性测定仪,Rapid Visco analyser,RVA）间接测定,但用淀粉谱仪测定的是淀粉的成糊温度（pasting temperature,PT）,它是指淀粉黏度刚开始升高时的温度。在许多情况下,从RVA配套软件 Thermocline for Windows中测定的PT是过高的。提出了两种能够用RVA较准确测定成糊温度的方法。一种是改变参数设置来“筛选”出能反映真正成糊温度的点;另一种是手工记录下淀粉黏度刚开始升高时的时间,利用公式(45/3.8)×(T1-1)+50 计算出稻米的成糊温度。利用手工计算出的成糊温度与通过差示扫描量热仪测定的最高糊化温度呈极显著正相关(r=0.97)。     

Accurate Measurement of Pasting Temperature by the Rapid Visco-Analyser： a Case Study Using Rice Flour

Pasting properties are among the most important characteristics of starch, determining its applications in food processing and other industries. Pasting temperature derived from the Rapid Visco-analyser (RVA) (Newport Scientific), in most cases, is overestimated by the Thermocline for Windows software program. Here, two methods facilitating accurate measurement of pasting temperature by RVA were described. One is to change parameter setting to ‘screen’ the true point where the pasting viscosity begins to increase, the other is to manually record the time (T1) when the pasting viscosity begins to increase and calculate the pasting temperature with the formula of (45/3.8)×(T1–1)+50 for rice flour. The latter method gave a manually determined pasting temperature which was significantly correlated with the gelatinization temperature measured by differential scanning calorimetry.