Gene Expression of Enzymes for Starch and Sucrose Metabolism and Transport in Leaf Sheaths of Rice (Oryza sativa L.) during the Heading Period in Relation to the Sink to Source Transition

Abstract

Leaf sheaths of rice plants are known to temporarily accumulate starch prior to heading, which is subsequently remobilized and transported into the panicle after heading. We investigated the time course for both carbohydrate content and steady state mRNA levels of enzymes related to starch and sucrose metabolism in the rice leaf sheath (Oryza sativa L. cv. Nipponbare). Leaf sheaths from the second leaf below the flag leaf accumulated high levels of starch before heading but they rapidly decreased after heading. In contrast, the flag leaf sheath did not accumulate as much starch. In the second leaf sheath, the mRNA levels of enzymes involved in starch synthesis, ADP glucose pyrophosphorylase (EC 2. 7. 7. 27), soluble starch synthase (EC 2. 4. 1.21) and branching enzyme (EC 2. 4. 1. 18) were high before heading, which coincided with rapid accumulation of starch. The mRNA levels of sucrose synthesis enzymes, cytosolic FBPase (EC 3. 1. 3. 11) and sucrose phosphate synthase (EC 2. 4. 1. 14), and the sucrose transporter (OsSUTI) increased at the time of heading, which was largely coincident with a decrease in the mRNA levels of starch synthesis enzymes. In the flag leaf sheaths, changes in mRNA levels of starch synthesis enzymes were not pronounced, however mRNA levels of sucrose synthesis enzymes and the sucrose transporter showed a clear increase throughout the heading period. The different characteristics observed between the two leaf sheaths will be discussed in relation to the sink to source transition.     

The Effect of the Amount of Nitrogen Fertilizer on Starch Metabolism in Leaf Sheath of Japonica and Indica Rice Varieties during the Heading Period

Abstract

The effects of the amount of nitrogen fertilizer on the starch metabolism of rice leaf sheath during the heading period in the japonica rice variety, cv. Nipponbare were compared with those in the indica varieties, cv. Tetep and Johna. The rice plants were grown under a low- (similar to the standard nitrogen level in paddy field) or high-nitrogen condition, and the starch content of the second leaf sheaths below the flag leaf was analyzed from the second leaf stage (growth stage 1) until 21 days after the heading (growth stage 7). The starch content of the plants grown under the high-nitrogen condition at the heading stage (growth stage 4) was lower than that under a low-nitrogen condition in all the varieties. The decrease in the activity of starch branching enzyme (SBE) was considered to be important for the repression of starch accumulation under a high-nitrogen condition. Under the high-nitrogen condition, Nipponbare accumulated more starch in the second leaf sheath than indica varieties at the heading stage. However, the phenomenon could not be accounted for by the activities of AGPase and SBE. Semi-quantitative RT-PCR analysis suggested that the lower activities of SBE in the second leaf sheath under the high-nitrogen condition may be due to, at least in part, the decrease in the expression level of RBE4.     

叶肉细胞特异表达焦磷酸化酶基因OsIP1在不同源库类型水稻品种中的效应

 将水稻自身焦磷酸化酶基因OsIP1置于叶肉细胞特异表达启动子下,利用农杆菌介导法将嵌合基因cyFBPase:OsIP1分别导入“源限制型” 水稻品种中超123和“库限制型” 品种农垦57。 根据荧光定量PCR结果和农艺性状,筛选高表达且农艺性状没有明显改变的T2转基因纯合株系,用以检测和分析在水稻叶肉细胞中特异性过表达水稻焦磷酸化酶基因的效应。初步的研究结果显示：1）在营养生长期,转基因株系的最高茎蘖数和穗数比各受体亲本均有不同程度的提高,尤其是在分蘖性较强的品种上;2）在籽粒灌浆结实期,叶片、叶鞘中可溶性总糖、蔗糖和淀粉含量在整个灌浆期的变化趋势与对照相似,且多数转基因植株叶片和叶鞘中的蔗糖含量（除了灌浆高峰期时的叶鞘）均显著高于对照;3）转基因株系单株干物质量较野生型对照有显著提高。单株产量呈现不同程度的增加,其中“源限制型”品种中超123转基因株系单株产量较对照增幅达显著水平。     

ABA对水分胁迫下水稻籽粒灌浆的调节

 利用水平分根系统研究了水稻抽穗后水分胁迫条件下ABA对水稻品种汕优63叶片光合特性、糖代谢和籽粒灌浆过程的调节。当一半根系受干旱胁迫时，叶片的水势和相对含量变化很小，但叶片和籽粒内ABA的含量显著增加。整个根系受干旱胁迫时，叶片的水势和相对含水量随着胁迫的增加而显著地降低，ABA含量则显著增加。半干和全干处理的水稻叶片的光合速率和RuBP羧化酶／氧化酶（Rubisco）的活性显著下降。叶片内蔗糖和淀粉的积累和代谢与叶片内ABA的含量和水分状况存在密切的关系，低浓度的ABA和高的叶水势与高的蔗糖积累和蔗糖磷酸合成酶活性相一致，但当叶片ABA的浓度增加时，即使叶片的水势与对照相似，叶片的蔗糖和淀粉积累以及蔗糖磷酸合成酶的活性则显著地下降，叶片的α－和β－淀粉酶活性似与ABA和水分状况关系不密切。ABA对籽粒灌浆有显著的抑制作用，ABA对籽粒灌浆的抑制作用至少部分与其对催化水稻籽粒内蔗糖代谢和淀粉积累的关键酶蔗糖合成酶的抑制有关。另外，干旱处理对叶片光合速率和蔗糖与淀粉积累的抑制程度显著高于半干旱处理，说明干旱本身也对这些过程有直接的效应。根据这些结果，作者认为，当水稻遇到干旱胁迫时，ABA对其光合作用、糖代谢以及籽粒的发育过程都有极其显著的调节作用。还就ABA对这些生理过程调节的可能性进行了讨论     

Development of introgression lines of an Indica-type rice variety, IR64, for unique agronomic traits and detection of the responsible chromosomal regions

A total of 334 introgression lines (INLs: BC₃-derived lines) derived from crosses between a recurrent parent of Indica rice cultivar IR64 and 10 donor parents, including new plant type (NPT) lines IR65600-87-2-2-3, IR65598-112-2, IR65564-2-2-3, IR69093-41-2-3-2, IR69125-25-3-1-1, Hoshiaoba, IR66215-44-2-3, IR68522-10-2-2, IR71195-AC1, and IR66750-6-2-1, have been developed. These INLs with IR64 genetic background were characterized for eight agronomic traits: days to heading, culm length, leaf width, leaf length, panicle length, panicle number, 100-grain weight, and total spikelet number per panicle at the International Rice Research Institute from 2005 to 2007. To identify introgressed segments from the donor parents, genotypes of the 334 INLs were detected using more than 200 polymorphic simple sequence repeat markers. These segments detected on chromosomes 1, 2, 4, 5, and 6 were commonly introgressed across the INLs from more than four donor varieties. Based on the data of phenotype and genotype for the 334 INLs, associations between agronomic traits and introgressed chromosomal segments in the 334 INLs were investigated. A total of 54 regions for the eight traits were detected: seven regions for days to heading, eight regions for culm length, eight regions for leaf width, four regions for leaf length, six regions for panicle length, three regions for panicle number per plant, seven regions for 100-grain weight, and 11 regions for total spikelet number per panicle. Among them, the region on the long arm of chromosome 4 was associated with characteristics of the NPT such as long leaf, broad leaf, and high spikelet number. The developed 334 INLs with the IR64 genetic background will be useful materials for genetic analysis of agronomic traits.     

外源赤霉素调控胡椒花穗形成及增产机理

花穗数目是影响胡椒产量的关键因素之一,前人研究发现喷施赤霉素可以增加胡椒花穗数目,但是增穗机理尚不明确.为了揭示赤霉素调控植物由营养生长向生殖生长转变的机理,选用幼龄胡椒为试材,设置不同浓度赤霉素(20、40、60、100 mg/L)和赤霉素抑制剂多效唑(1500 mg/L),以清水为对照,进行喷施试验.研究测定胡椒的...     

两种生长调节剂对大豆叶片昼夜同化物生理代谢及产量的影响

为探究两种不同类型植物生长调节剂对大豆叶片昼夜同化物生理代谢的影响,以两个大豆品种合丰50和垦丰16为试验材料,于始花期叶面喷施烯效唑(S₃₃₀₇)和2-N,N-二乙氨基乙基己酸酯(DTA-6),在鼓粒期的一昼夜内依次选取9∶30、17∶30、21∶30和5∶30(次日)4个时间点,测定功能叶片中蔗糖含量、淀粉含量、淀粉转化率、蔗糖代谢关键酶活性,并于成熟期测产。结果表明:S₃₃₀₇和DTA-6处理均能促进两品种叶片蔗糖和淀粉在白天的积累和夜间的降解,且S₃₃₀₇对两品种叶片蔗糖的调控效果优于DTA-6,DTA-6调控两品种叶片淀粉的作用效果优于S₃₃₀₇。在不同时间点S₃₃₀₇和DTA-6处理均能够提高两品种叶片夜间蔗糖和淀粉转运率。S₃₃₀₇和DTA-6处理能够提高两品种叶片蔗糖合酶和蔗糖磷酸合酶活性,且均在17∶30达最大值;S₃₃₀₇和DTA-6在整个昼夜变化中提高两品种蔗糖转化酶的活性。S₃₃₀₇和DTA-6能够有效调控两品种的单株荚数和单株粒数,提高产量,S₃₃₀₇和DTA-6处理后合丰50产量与对照相比分别增加17.76%和37.67%,垦丰16产量与对照相比分别增加11.34%和32.49%。综上所述,S₃₃₀₇和DTA-6处理均能通过促进叶片昼夜同化物代谢,提高合丰50和垦丰16大豆品种产量。     

Carbohydrate metabolism during tuber initiation in potato: A transient surge in invertase activity marks the stolon to tuber transition

Summary The relationship between carbohydrate metabolism and tuber initiation in potato was determined by monitoring changes in the amount of starch and sugars along with the activities of sugar metabolizing enzymes upon transfer of plants to tuber-inducing conditions (TI; short days, cold nights) from non-inducing conditions (NTI: long days, warm nights). Switch to TI conditions caused an immediate slow-down in plant growth and triggered swelling of stolon tips, which went on to develop into tubers. Leaves of plants moved to TI conditions accumulated less starch and sugar while their stolon tips showed a sudden upsurge in starch content and a sharp decline in sugars even before any tip swelling was detectable. These changes were paralleled by a transient surge in the activity of cell wall invertase (74%) and soluble invertase (30%) in stolon tips of plants transferred to TI conditions in two unrelated cultivars under different experimental conditions. As the surge in invertase activity faded, it was replaced by a substantial increase in sucrose synthase activity as the tuber enlargement proceeded. The transient increase in invertase activity just prior to tuber initiation appears to mark a turning point in the transition of stolon tip to tuber.     

自然光温和控制光温下玉米苗期叶片NSC比较及根系糖代谢日变化

通过对比玉米苗期叶片在田间和室内条件下NSC日变化特征,分析自然条件下光照和温度变化与光合碳同化产物间的关联性,对室外水培条件下玉米苗期根系蔗糖、总可溶性糖和淀粉日变化特性进行探讨。结果表明,玉米苗期叶片NSC累积呈先升高后降低的特征,不同处理NSC峰值均出现在光照结束阶段;田间光温条件下淀粉和可溶性糖累积浓度均高于室内水培。光温变化对NSC积累的影响达极显著水平(P0.01);前一时间点的光照与后一时间点的淀粉和可溶性总糖之间相关性均达极显著水平(R2=0.77;R2=0.90)。根系中可溶性糖浓度呈明显的日变化特征,淀粉则在141.9～161.9 mg/g DM范围内波动。     

水稻拔节期叶片碳氮代谢基因型差异及与氮素利用效率的关系

 采用土培盆栽试验,以3个氮素利用效率(NUE)有显著差异的水稻基因型秀恢2号(低NUE)、R83 12(中NUE)和五优244(高NUE)为材料,研究了拔节期叶片碳氮代谢的基因型差异及其与氮素高效利用的关系。结果表明,拔节期不同基因型水稻的叶片蔗糖和可溶性糖含量、淀粉/可溶性糖含量比和淀粉/蔗糖含量比存在极显著差异,前两者为五优244 > R83 12 >秀恢2号,后两者则相反;而淀粉和蔗糖/可溶性糖含量比各基因型差异不显著。不同基因型水稻的叶片全氮含量(TNC)、蛋白氮含量(PNC)、 PNC/TNC、Rubisco含量(RC)/PNC和RC/TNC存在极显著或显著差异,前两者为五优244 < R83 12 <秀恢2号, 后三者则相反;RC无基因型差异。五优244叶片的叶绿素a含量、类胡萝卜素含量、PSⅡ潜在活性、羧化效率、碳酸酐酶、酸性转化酶和蔗糖磷酸合成酶活性极显著或显著高于其他基因型;其叶片的硝酸还原酶、谷氨酰胺合成酶、谷氨酸合成酶和谷氨酸脱氢酶活性也极显著高于其他基因型。统计分析表明,高NUE水稻叶片具有较高的可溶性糖和蔗糖含量及较大的RC/TNC和RC/PNC,其生理基础是其叶片的高谷氨酰胺合成酶活性和PSⅡ潜在活性。     

不同播期下优质稻桂华占、八桂香花后碳氮物质流转与籽粒生长相关性

以桂华占、八桂香为材料,不同播期调控下,研究不同播期下优质稻花后植株碳氮流转与籽粒生长及品质的相关性.结果表明:(1)花后茎鞘、叶片干物质运转速度和运转率都与籽粒起始灌浆势呈正相关.籽粒活跃灌浆期、持续灌浆时间与花后茎鞘、叶片干物质运转速度和运转率呈极显著正相关.(2)播种期推迟不利于茎鞘碳同化物向穗部流转,茎鞘碳同化物转运对籽粒的产量和淀粉产量的贡献率表现为淀粉＞可溶性糖＞蔗糖,茎鞘碳同化物对籽粒产量及淀粉产量的贡献率远高于叶片.可溶性糖转运对籽粒产量和淀粉产量贡献率表现为SD1＞SD2＞SD3;蔗糖、淀粉对籽粒的产量贡献率表现为SD1＞SD2＞SD3.茎叶可溶性糖积累量的减少与籽粒直链淀粉含量和积累量增加是同步的,并且,茎叶可溶性糖积累量快速递减期(花后3～12 d)与直链淀粉含量和积累量快速递增期(花后6～12d)同步.(3)播种期推迟减少茎鞘和叶片总氮的积累,籽粒氮收获指数降低,但是播种期的推迟却增加茎鞘和叶片器官蛋白氮积累,有利于籽粒蛋白质含量提高.     

Effect of various drought stresses and subsequent recovery on proline,total soluble sugar and starch metabolisms in Rice (Oryza sativa L.) varieties

ABSTRACT

Rice (Oryza sativa L.) is one of the most important staple foods in the world, however most improved rice varieties are susceptible to drought stress. A two-year study was conducted to explore the effects of various drought stresses and subsequent recovery on the accumulation and degradation of proline, total soluble sugar and starch in different rice varieties at vegetative stage. The results showed that relative water content in the leaves and sheaths of rice varieties significantly decreased under drought stresses, but not at the same rate. Under control and drought conditions, the water content in sheaths was higher than that in leaves. Interestingly, under severe drought stress in 2015, the leaf water content was higher than the sheath water content. The water distribution between leaves and sheaths might be a response of plants to protect leaf system from devastation by drought. Proline was highly accumulated under drought stress but rapidly decreased after re-watering. The drought tolerant variety DA8 expressed higher ability in accumulation of proline than susceptible varieties. In general, total soluble sugar and starch contents in leaves and sheaths of varieties decreased under drought stress conditions. Total soluble sugar and starch content of DA8 were less affected than other varieties under drought conditions. Our study indicated that metabolisms of total soluble sugar and starch in rice were affected by both environmental conditions and characteristics of varieties. Proline accumulation ability of varieties can be used as a useful indicator for drought tolerant potential in rice breeding for water-limited environments.     

灌溉对弱筋小麦豫麦50籽粒淀粉积累及其相关酶活性的影响

为了探明灌溉对淀粉积累及其相关酶活性的影响规律,在大田条件下,以弱筋小麦豫麦50为材料,研究了三种灌溉方式(W1:拔节期灌水一次;W2:拔节期和孕穗期各灌水一次;W3:拔节期、孕穗期和灌浆期各灌水一次.每次灌水定额750 m3/ha)下弱筋小麦淀粉及其相关酶活性的变化规律.结果表明,弱筋小麦灌浆中后期和成熟期籽粒直链淀粉、支链淀粉、总淀粉积累速率均表现为W3>W2>W1.成孰期籽粒淀粉支/直比例表现为W3＞W2＞W1.成熟期籽粒淀粉支/直比例表现为W3＜W2＜W1.孕穗期和灌浆期增加灌水提高了灌浆中后期旗叶和籽粒中蔗糖含量、蔗糖合成酶(SS)活性、蔗糖磷酸合成酶（SPS）活性,提高了灌泺中后期籽粒中可溶性淀粉合成酶(SSS)活性和束缚态淀粉合成酶(GBSS)活性.说明增加灌水使弱筋小麦旗叶蔗糖合成能力强,促进源器官蔗糖向库中运输,保证籽粒库中糖源的充分供应,有利于籽粒淀粉的合成和积累.     

玉米C4光合叶不同部位“花环”结构及叶绿素含量的变化

以玉米进行C4光合的全展第5位叶片为材料,分析从叶基部到顶部的解剖结构和叶绿素含量变化,研究玉米C4光合叶片"花环"结构随叶片发育的变化规律。结果表明,玉米第5位叶从基部到顶部都具有完整的典型"花环"结构,维管束鞘细胞(BSC)和叶肉细胞(MC)的体积在叶片发育过程中具有渐变性,从叶基部到顶部BSC和MC均呈先增大后变小的趋势,而且叶绿素a、b和a+b含量呈相同的变化趋势,说明BSC和MC细胞体积与叶绿素含量的变化具有相关性。叶绿素a/b总体呈上升趋势,说明玉米第5叶基部到顶部的光合途径存在C3向C4转变的过程。玉米第5叶不同部位C4光合途径发育的渐变性比前3叶更为明显。     

Growth rate and carbohydrate metabolism of potato tubers exposed to high temperatures

Summary The effect of high tuber temperature (30 °C) on the growth rate and carbohydrate metabolism of tubers was studied with 3 cultivars differing in heat tolerance. Subjecting individual tubers to 30°C for 6 days caused cessation of tuber growth. During this period, non-treated tubers (20°C) on the same plant increased in volume. At 30°C, incorporation of¹⁴C-labelled assimilates into the ethanol-insoluble fraction (mainly starch) as well as the starch content was significantly reduced. In contrast, the incorporation of¹⁴C-labelled assimilates into the sugar fraction was not affected by high tuber temperature. At 30°C the activity of some of the enzymes involved in starch metabolism was depressed. Varietal differences in responses of tuber growth rate and carbohydrate metabolism to high tuber temperature were insignificant. From these preliminary results it is concluded that high soil temperatures may decrease starch content of potato tubers directly by inhibition of the conversion of sugars into starch.     

外源二乙基二硫代氨基甲酸钠 对花期淹水大豆碳代谢的影响

      为了研究二乙基二硫代氨基甲酸钠（DDTC）对大豆碳水化合物代谢的调节作用,为大豆抗渍涝栽培提供技术支持,本研究以渍涝敏感型大豆品种徐豆18和耐渍涝品种南农1138-2为材料,采用盆栽的方式,花期淹水前喷施DDTC,调查大豆叶片光合特性、蔗糖代谢酶活性及叶片、根系和根瘤中碳水化合物含量。结果表明：淹水胁迫降低了植株地上部、根系和根瘤干物重,叶片净光合速率（Pn）、气孔导度（Gs）、胞间二氧化碳浓度（Ci）、蒸腾速率（Tr）,根系中淀粉含量、根瘤中蔗糖和淀粉含量。其中,南农1138-2植株干物重,叶片光合气体交换参数以及根系中淀粉含量比徐豆18下降幅度小,而两品种间根瘤中蔗糖和淀粉含量下降幅度基本一致。与之不同,叶片中蔗糖磷酸合成酶（SPS）、蔗糖合成酶（SS）（合成方向）活性、SS（分解方向）、酸性转化酶（AI）活性,叶片可溶性糖、蔗糖和淀粉含量,根系中可溶性糖、蔗糖含量增加。其中,南农1138-2叶片中蔗糖合成酶（SS）（分解方向）和酸性转化酶（AI）活性、可溶性糖、蔗糖和淀粉含量增幅较小,而叶片中蔗糖磷酸合成酶（SPS）、SS（合成方向）活性增幅较大。DDTC处理提高了淹水胁迫下两品种植株干物重,缓解了淹水胁迫对大豆植株生长的抑制;提高了叶片光合气体交换参数,叶片中SPS和SS（合成方向）活性,并降低了SS（分解方向）和AI活性,促进了淹水胁迫下叶片中蔗糖向根系和根瘤的转运;DDTC处理也增加了根系中可溶性糖、蔗糖和淀粉含量,根瘤中蔗糖和淀粉含量,表明DDTC处理在调控淹水胁迫下大豆植株生长和碳代谢生理过程中起着重要的作用。     

氮素用量对春玉米穗位叶蔗糖合成关键酶活性的影响

试验以高淀粉玉米四单19、普通玉米东农250、优质蛋白玉米丰禾10为材料,研究氮素用量对春玉米穗位叶蔗糖代谢的影响,揭示蔗糖合成过程中蔗糖合成酶(SS)和蔗糖磷酸合成酶(SPS)的调节作用。结果表明：氮素用量适当有利于提高玉米穗位叶蔗糖含量、蔗糖合成酶活性和蔗糖磷酸合成酶活性,证实了蔗糖合成酶和蔗糖磷酸合成酶能促进春玉米叶片蔗糖积累。     

Storage and Remobilization of Soluble Carbohydrates after Heading in Different Plant Parts of a Winter Wheat Cultivar

Abstract

Several types of water soluble carbohydrates (WSG) were traced into plant parts of a winter wheat (Triticum aestivum L.) cultivar from heading to physiological maturity. Field grown plants were harvested at intervals of a few days and divided into grain, chaff, leaf blades, leaf sheaths, rachis + peduncle and culm internodes. Leaf blades and sheath showed the peak of WSC contents at about anthesis. Culm internodes accumulated fructan and sucrose during the early grain-filling phase, from a week after anthesis until the milk-ripe stage, then remobilized them during late and final grain filling phases, from the milk-ripe stage until maturity. The amount of sucrose known as short-term storage WSC was higher than fructan known as long-term storage WSC in each internode throughout the grain-filling phase. Chaff showed a large amount of fructan and fructose before anthesis, although it did very little sucrose. A pattern of sucrose accumulation in chaff was very different from that of fructan, unlike the other parts. These patterns of changes in WSC contents in plant organs roughly corresponded with four grain-filling phases, the initial, early, late and final phases.     

Structural and Functional Differentiation of Bundle Sheath and Mesophyll Cells in the Lamina Joint of Rice Compared with that in the Corresponding Region of the Liguleless Genotype

Abstract

The structural and functional characterization of the blade-sheath boundary region of a rice cultivar T65 and its near-isogenic line T65lg were examined by light and electron microscopy and in situ hybridization. Starch accumulation in bundle sheath cells was compared between the lamina joint of T65 and the corresponding region of T65lg and also between the lamina joint and the leaf blade. In the lamina joint of T65, starch grains were predominantly accumulated in bundle sheath cells, and the starch-containing chloroplasts within these cells were spherical in shape. On the other hand, in the blade-sheath transition region of T65lg, little starch accumulation was observed and the chloroplasts were oval in both mesophyll and bundle sheath cells. Furthermore, photosynthesis-related genes, rbcS and cab, were expressed in mesophyll cells within the blade-sheath transition region of T65lg as in the leaf blade and sheath, while no expression of these genes was found within the lamina joint of T65. These facts indicate that T65lg can not develop the lamina joint from either structural or functional aspect. The present results suggest that the control mechanism of starch accumulation in bundle sheath chloroplasts in the lamina joint differs from that in leaf blade in rice.