Changes in Enzyme Activities Involved in Starch Synthesis and Hormone Concentrations in Superior and Inferior Spikelets and Their Association with Grain Filling of Super Rice

The changes in activities of key enzymes involved in sucrose-to-starch conversion and concentrations of hormones in superior and inferior spikelets of super rice were investigated and their association with grain filling was analyzed.Four super rice cultivars,Liangyoupeijiu,IIyou 084,Huaidao 9 and Wujing 15,and two high-yielding and elite check cultivars,Shanyou 63 and Yangfujing 8,were used.The activities of sucrose synthase (SuSase),adenosine diphosphoglucose pyrophosphorylase (AGPase),starch synthase (StSase) and starch branching enzyme (SBE),and the concentrations of zeatin + zeatin riboside (Z + ZR),indole-3-acetic acid (IAA) and abscisic acid (ABA) in superior and inferior spikelets were determined during the grain filling period and their relationships with grain filling rate were analyzed.Maximum grain filling rate,the time reaching the maximum grain-filling rate,mean grain filling rate and brown rice weight for superior spikelets showed a slight difference between the super and check rice cultivars,but were significantly lower in the super rice than in the check rice for inferior spikelets.Changes of enzyme activities and hormone concentrations in grains exhibited single peak curves during the grain filling period.The peak values and the mean activities of SuSase,AGPase,StSase and SBE were lower in inferior spikelets than in superior ones,as well as the peak values and the mean concentrations of Z + ZR and IAA.However,the peak value and the mean concentration of ABA were significantly higher in inferior spikelets than in superior ones and greater in the super rice than in the check rice.The grain filling rate was positively and significantly correlated with the activities of SuSase,AGPase and StSase and the concentrations of Z + ZR and IAA.The results suggested that the low activities of SuSase,AGPase and StSase and the low concentrations of Z + ZR and IAA might be important physiological reasons for the slow grain filling rate and light grain weight of inferior spikelets in super rice.     

施氮量对小麦籽粒灌浆的影响及其生理机制

为探明施氮量对小麦籽粒灌浆的影响及其生理机制,以当地高产小麦品种扬麦16和宁麦13为材料,设置120 kg·hm^-2(LN)、180 kg·hm^-2(MN)和240 kg·hm^-2(HN) 3个施氮量处理,分析了在不同施氮量下,强、弱势籽粒灌浆特征与籽粒中蔗糖-淀粉代谢途径中关键酶活性的变化。结果表明,两个小麦品种的籽粒产量随着施氮量的增加而增加,但在MN与HN处理间差异不显著。氮肥偏生产力随着氮肥用量的增加而显著降低。与LN处理相比,MN处理提高了扬麦16弱势粒中腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)、可溶性淀粉合成酶(SSS)和淀粉分支酶(SBE)的活性、弱势粒灌浆速率和粒重;HN则降低了扬麦16弱势粒中被测酶的活性和粒重。宁麦13弱势粒中AGPase、SSS和SBE活性、弱势粒灌浆速率和粒重则随施氮量的增加而降低。施氮量对两个小麦品种强势粒中被测酶活性、籽粒灌浆速率和粒重无显著影响。以上结果说明施氮量主要是影响小麦弱势粒的灌浆,且在品种间存在差异;施用氮肥后弱势粒灌浆速率和粒重的增加或降低与籽粒中蔗糖-淀粉代谢途径中关键酶活性的增强或降低密切相关;在地力较好条件下,施氮量为180 kg·hm^-2时就可以获得高产与氮肥高效利用的效果。     

不同冠温特征小麦的籽粒灌浆特性及内源激素的变化

为阐明小麦冠层温度与其籽粒灌浆特性、内源激素变化的关系,2001~2003年对黄淮麦区及河南省推广的30个小麦品种冠温特征、籽粒灌浆特性和籽粒发育过程中内源激素(玉米素ZR、赤霉素GA3、生长素IAA、脱落酸ABA)含量及比值的变化进行了研究。结果表明,豫麦50在灌浆后期冠层温度明显降低,表现为冷尾型;豫麦34和豫麦70在灌浆后期冠层温度有上升的趋势,表现为暖尾型。在灌浆末期冷尾型与暖尾型小麦冠层温度相差超过2.5℃。冷尾型豫麦50强、弱势粒的最大灌浆速率、平均灌浆速率和渐增期、快增期、缓增期的灌浆速率均大于暖尾型豫麦34、豫麦70;强势粒的激素含量变化及其比值变化规律性明显,弱势粒表现的规律性不如强势粒。冷尾型豫麦50强势粒ZR、IAA、GA3含量峰值均高于豫麦34、豫麦70,其ZR/ABA、GA3/ABA、IAA/ABA峰值亦高于暖尾型豫麦70和豫麦34,而ABA含量在灌浆中期上升快,且高峰值大于豫麦70和豫麦34,达到高峰后下降较快;在弱势粒中,豫麦50上述各种激素含量仍保持较高状态。     

灌浆期低温对离体玉米子粒灌浆过程及内源激素的影响

以郑单958为材料,采用子粒离体培养方法,研究灌浆期低温对玉米强、弱势粒灌浆特性及内源激素的影响。研究表明,灌浆期低温分别减缓强、弱势粒平均灌浆速率15%、23%,延长灌浆时间约8 d,最终粒重分别降低3%、9%。低温处理下,GA_3、IAA、ZR含量与灌浆速率显著相关;受低温影响,强、弱势粒的激素含量变化趋势相近,与常温相比,子粒GA_3含量先增加后减少,IAA、ZR含量低于对照,且对弱势粒的影响大于强势粒。低温对内源激素含量的影响是导致灌浆速率降低的主要原因之一。     

6-BA对玉米子粒内源激素含量的影响

在授粉前喷施60 mg/L的6-BA,对玉米胚乳发育过程中主要内源激素(Z+ZR、IPA、IAA、ABA)含量的差异及其动态变化差异作了比较分析。结果表明,外源6-BA对胚乳细胞增殖的影响主要表现为加快了胚乳细胞增殖速率,提前胚乳细胞最大增殖速率出现的时间。在玉米灌浆初期,6-BA使胚乳内Z+ZR、IPA、IAA含量明显高于对照,ABA含量低于对照,可能是胚乳细胞数目增加的主要原因。     

IAA与亚种间杂交稻籽粒发育的关系及烯效唑的调节

 以汕优413及其父本品种中413为材料,研究了内外源IAA在籽粒发育过程中的作用及植物生长延缓剂S-07对其强、弱势粒灌浆过程的影响。在灌浆前期,强势粒抑制弱势粒的发育,外源IAA可代替强势粒的这种抑制作用;灌浆过程中强、弱势粒内源IAA的变化动态存在差异,其中强势粒的内源IAA在开花后急剧上升,8 d左右达最高峰;而弱势粒的IAA水平开花后则有一段约12 d的“滞缓期”,在开花后20 d左右才有一高峰出现。强弱势粒对3H-IAA的合成和吸收能力差异可能是导致其内源IAA水平不平衡的原因之一。孕穗末期喷施S-07可部分抑制强势粒的发育而相对促进弱势粒的发育,提高弱势粒千粒重;孕穗末期喷施S-07溶液可降低强势粒但提高弱势粒灌浆前期的内源IAA水平。     

植物激素对杂交稻籽粒灌浆及蔗糖合酶活性的影响

 以亚种间杂交稻两优培九和品种间杂交稻汕优63为材料,对籽粒灌浆期间的内源激素含量和酶活性进行了测定分析,并结合外源激素对水稻籽粒灌浆过程的调控试验,分析了脱落酸对蔗糖代谢关键酶——蔗糖合酶活性的调节作用。结果表明,灌浆前期强势粒中生长素、玉米素核苷和脱落酸（ABA）含量明显高于弱势粒,并与籽粒最大灌浆速率、平均灌浆速率、籽粒充实率和粒重均呈显著或极显著正相关。强势粒中蔗糖合酶活性明显高于弱势粒,且达到峰值的时间也早于弱势粒。籽粒灌浆速率、籽粒充实率、粒重和淀粉积累速率与蔗糖合酶活性呈显著正相关。花后8 d对稻株喷施低浓度ABA显著提高了籽粒中ABA含量和蔗糖合酶活性,增加了粒重和籽粒充实率。喷施ABA合成抑制剂氟草酮作用则相反。上述结果说明,植物激素尤其是ABA对水稻籽粒灌浆有重要的促进作用,其作用在很大程度上是通过调节籽粒蔗糖合酶活性实现的。     

亚种间杂交水稻灌浆特性的调节及其对产量的影响

 以亚种间组合汕优413和协优413、籼型杂交组合汕优10号为材料进行研究。结果表明对弱势粒灌浆的影响,IAA具有代替强势粒的作用。即使剪去顶端3个一次枝梗、营养物供应非常充足,由于IAA的存在,弱势粒的灌浆也较为迟缓,最终形成大量空秕粒,与不修剪的对照接近,说明弱势粒的灌浆主要受IAA调控,也证明强势粒以“顶端优势”的方式抑制弱势粒的灌浆。     

内源IAA对杂交稻强、弱势粒灌浆增重的影响

以超级杂交稻协优9308和杂交稻协优9312、Ⅱ优162及其相应恢复系为材料,对灌浆初期籽粒中内源IAA、糖分的变化,以及IAA抑制剂对IAA以及3H 标记葡萄糖吸收分配和籽粒灌浆的影响进行了研究。结果表明在籽粒灌浆初期强势粒的IAA含量以及增重的速度始终高于弱势粒。在开花受精7 d以前,强势粒中果糖、葡萄糖及蔗糖的含量高于弱势粒,之后相反。强、弱势粒间存在的这种灌浆势差异,杂交组合大于其相应的恢复系。IAA合成及运输抑制剂明显降低3H 葡萄糖在籽粒中的分配。外源IAA对幼穗的处理,缩小了强、弱势粒间结实率的差异,使整穗的结实率也有所提高。     

灌浆前期高温对小麦籽粒氮代谢关键酶活性的影响

为探讨花后高温胁迫对小麦籽粒氮代谢及蛋白质合成的影响机制,以黄淮地区高产小麦品种郑麦366为材料,利用人工气候室模拟花后高温的方式,在盆栽条件下研究了灌浆前期短暂高温对小麦强势粒和弱势粒谷丙转氨酶(GPT)、谷草转氨酶(GOT)活性的影响.结果表明,灌浆前期高温显著提高了籽粒蛋白质含量,降低了籽粒蛋白质积累量和氮代谢关键酶GPT、GOT活性,其中强势粒GPT活性受高温胁迫的影响较大.小麦籽粒蛋白质含量与GPT和GOT活性均呈负相关,但相关性在不同时期间、强势粒与弱势粒间有差异,强势粒GPT活性与蛋白质含量的关系更密切.     

Hormonal changes are related to the poor grain filling in the inferior spikelets of rice cultivated under non-flooded and mulched condition

Non-flooded plastic mulching cultivation (PM) of rice can save much irrigation but usually exhibits a poor grain filling and low grain weight when compared to traditional flooding cultivation (TF). This study measured the variations of plant hormones during grain filling and investigated whether they were related to the grain-filling problem under mulching in a field experiment. Hormonal levels in the grains and the grain development of both superior (early flowered) and inferior spikelets (later flowered) were monitored continuously during the grain filling period. The contents of indole-3-acetic acid (IAA), zeatin riboside (ZR) and abscisic acid (ABA) were higher in superior grains than in inferior ones at early grain-filling stage. For individual spikelets, the peaks of IAA and ZR contents appeared just before the peak grain-filling rate but the peak ABA content matched it. The earlier flowered superior spikelets did not show much difference in grain filling characteristics and hormonal changes under the two cultivation treatments. Distinctively with the inferior spikelets in a panicle, PM led to lower grain weight, higher grain-filling rate at early stage and shorter active grain-filling period than the TF. PM also resulted in less IAA and ZR but more ABA contents in the grains than TF at early and middle grain filling stages in these inferior spikelets. Peaks of these hormone contents in the PM inferior grains usually appeared earlier and also disappeared more quickly than those in TF inferior grains. When exogenous IAA was applied to plants at the initial grain-filling stage, IAA and ZR were increased and ABA was reduced in the inferior grains. The treatment prolonged the active grain-filing period and reduced the peak grain-filling rate of inferior grains, similarly as the case with TF. As a result, IAA spraying enhanced the weight of PM inferior grains but reduced that of TF ones. Exogenous ABA spraying led to the opposite effects to those of IAA. The results suggest that the shortened grain-filling period in the inferior grains under PM is related to the increased ABA and reduced IAA and ZR in the grains. Regulation of the ratio of ABA to IAA in grains could potentially increase the weight of inferior grains under the water-saving cultivation with film mulching.     

Pre-anthesis non-structural carbohydrate reserve in the stem enhances the sink strength of inferior spikelets during grain filling of rice

Sink strength plays an important role in grain filling of cereals but how it is related to the pre-anthesis non-structural carbohydrate (NSC) reserves is not clear. This study investigated if and how an increase in NSC reserves could enhance sink strength, and consequently improve grain filling of later-flowering inferior spikelets (in contrast to the earlier flowering superior spikelets) for rice varieties with large panicles. Two “super” rice varieties (the recently bred high-yielding rice) and two New Plant Type (NPT, named in IRRI for the extra-large panicle) rice lines were compared with two elite inbred varieties under field-grown conditions. Three nitrogen (N) treatments, applied at the stages of panicle initiation, spikelet differentiation or both, were adopted with no N application during the mid-season as control. Both super rice and NPT rice showed a greater yield capacity as a result of a larger panicle than the elite inbred rice. However, a lower percentage of filled grains limited the realization of higher yield potential in super rice and especially in NPT rice, due to their lower grain filling rate and the smaller grain weight of their inferior spikelets. The low grain filling rate and small grain weight of inferior spikelets are mainly attributed to a poor sink strength as a result of small sink size (small number of endosperm cells) and low sink activity, e.g. low activities of sucrose synthase (SuSase) and adenosine diphosphoglucose pyrophosphorylase (AGPase). The amounts of NSC in the stem and NSC per spikelet at the heading time are significantly and positively correlated with sink strength (number of endosperm cells and activities of SuSase and AGPase), grain filling rate, and grain weight of inferior spikelets. Nitrogen application at the spikelet differentiation stage significantly increased, whereas N application at the panicle initiation or at both panicle initiation and spikelet differentiation stages, significantly reduced, NSC per spikelet at the heading time, sink strength, grain filling rate, and grain weight of inferior spikelets in super rice. The results suggest that pre-anthesis NSC reserves in the stem are closely associated with the sink strength during grain filling of rice, and N application at the spikelet differentiation stage would be a good practice to increase pre-anthesis NSC reserves, and consequently to enhance sink strength for rice varieties with large panicles, such as super rice varieties.     

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。     

Dynamics of Abscisic Acid Levels during Grain-Filling in Rice : Comparisons between Superior and Inferior Spikelets

Abstract

In a rice panicle, the superior spikelets display higher growth rates than the inferior spikelets during the initial phase of grain-filling. To better understand the regulatory mechanism of this phenomenon, we examined the dynamics of endogenous abscisic-acid (ABA) levels and the effects of shading on the dynamics during the grain-filling period in superior and inferior spikelets. While ABA content in the superior spikelets increased rapidly after flowering, that in the inferior spikelets increased slowly and reached the maximum later than in the superior spikelets. Shading significantly exaggerated the inherently different ABA dynamics between the superior and inferior spikelets. We concluded that ABA levels are correlated with the dry matter accumulation patterns of spikelets. These results support our hypothesis that ABA is involved in assimilate partitioning among spikelets in a panicle during the initial phase of grain-filling of rice.     

杂交粳稻屉优418及其亲本灌浆期叶片内源激素含量变化与衰老之间的关系

以杂交水稻屉优418及其亲本屉锦和C418为材料,研究了生育后期叶片内源激素含量变化与衰老之间的关系。结果表明杂交水稻及其亲本剑叶中ABA、GA1/3、ZR和IAA含量变化趋势基本一致。3个材料的（GA1/3+ZR+IAA）/ABA的比值变化总的趋势也基本相同,前期较平稳,后期比值迅速下降。在生育后期,屉优418和C418的（GA1/3+ZR+IAA）/ABA比值下降速率较慢,与衰老相对较慢一致;而屉锦的（GA1/3+ZR+IAA）/ABA比值下降速率较快,与其衰老较快相一致。     

Relationship Between Changes in Leaf Endogenous Hormone Contents and Senescence During Grain Filling Stage of a Rice Hybrid and its Parents

A major problem in hybrid rice production is the occurrence of leaf senescence during the grain filling stage that can result in reduction of yield. Changes in contents of several endogenous hormones are related to leaf senescence. The relationship between endogenous hormones and leaf senescence in the rice hybrid Tiyou 418 and its parents Tijin and C418, was undertaken for investigation. Indicators of leaf senescence, including superoxide dismutase (SOD) activity, malondialdehyde (MDA) accumulation and chlorophyll content, as well as the contents of abscisic acid (ABA), zeatin riboside (ZR), gibberellin (GA1/3) and auxin (IAA) in the leaves were determined. Different rates of leaf senescence were observed in the three materials. Senescence occurred earliest and fastest in Tijin, followed by Tiyou 418 and then C418. A similar trend was recorded in ABA, ZR, and IAA contents during the grain filling stage in the three materials. Changes in (GA1/3 ZR IAA)/ABA ratios were also similar, being quite stable during the early stage of leaf senescence, and decreasing markedly during the late stage. The ratio declined more dramatically in Tijin, in accordance with its faster leaf senescence. The results suggest that the ratio of (GA1/3 ZR IAA)/ABA regulates chlorophyll content, SOD activity, MDA content and membrane lipid peroxidation. It is postulated that endogenous hormones may play a role in the regulation of leaf senescence in a systematic way.     

Relationship Between Changes in Leaf Endogenous Hormone Contents and Senescence During Grain Filling Stage of a Rice Hybrid and its Parents

A major problem in hybrid rice production is the occurrence of leaf senescence during the grain filling stage that can result in reduction of yield. Changes in contents of several endogenous hormones are related to leaf senescence. The relationship between endogenous hormones and leaf senescence in the rice hybrid Tiyou 418 and its parents Tijin and C418, was undertaken for investigation. Indicators of leaf senescence, including superoxide dismutase (SOD) activity, malondialdehyde (MDA) accumulation and chlorophyll content, as well as the contents of abscisic acid (ABA), zeatin riboside (ZR), gibberellin (GA1/3) and auxin (IAA) in the leaves were determined. Different rates of leaf senescence were observed in the three materials. Senescence occurred earliest and fastest in Tijin, followed by Tiyou 418 and then C418. A similar trend was recorded in ABA, ZR, and IAA contents during the grain filling stage in the three materials. Changes in (GA1/3 ZR IAA)/ABA ratios were also similar, being quite stable during the early stage of leaf senescence, and decreasing markedly during the late stage. The ratio declined more dramatically in Tijin, in accordance with its faster leaf senescence. The results suggest that the ratio of (GA1/3 ZR IAA)/ABA regulates chlorophyll content, SOD activity, MDA content and membrane lipid peroxidation. It is postulated that endogenous hormones may play a role in the regulation of leaf senescence in a systematic way.