Mobilization of metabolites from leaves to grains as the cause of monocarpic senescence in rice

The pattern of senescence was studied by following the changes in chlorophyll and protein in the leaves and by measuring ³²P retention and export from source to sink during development of the rice plant (Oryza sativa L. cv. Jaya) subjected to different manipulative treatments. With the advance of reproductive development, the chronological sequence of leaf senescence was changed, so that the flag and the third leaf senesced earlier than did the second leaf. In presence of the daughter shoot of defruited plants, senescence was delayed in all three leaves of the mother plant, as compared to the same leaves of intact plants. Senescence of all three leaves was further delayed when both panicle and daughter shoots were removed from the plant. The above manipulative treatments caused the initial sequential pattern of senescence of leaves to persist. Removal of both panicle and daughter shoots caused little export of ³²P between leaves. In the presence of daughter shoots of defruited plants, export of ³²P was maximum from leaves of the mother plant to the nearest daughter shoots. This led to earlier senescence of such mother plant leaves than that of plants from which both panicle and daughter shoots were removed. The pattern of senescence and export of ³²P in the flag and the second leaf of the daughter shoot was essentially the same as that of the intact plant. Based on these findings, it was concluded that mobilization of metabolites from source to sink is the primary cause of monocarpic senescence in rice.     

Monocarpic senescence in wheat: Influence of sterile glumes and ear

The senescence of sterile glumes, flag leaf and the other two leaves below the ear of wheat ( Triticum aestivum L. cv. Sonalika) was studied in relation to grain development and surgical manipulation. The senescence of sterile glumes was faster than that of the leaves in terms of chlorophyll and protein degradation. The flag leaf senesced later than the other two leaves below it. Removal of sterile glumes markedly reduced the harvest index (crop: straw ratio) and average dry weight per grain as compared to removal of the flag leaf. Maximum grain weight was achieved after the glumes had senesced completely. Removal of the ear delayed senescence of all the three leaves. It is concluded that sterile glumes are important suppliers of assimilate for grain filling and that nutrient drainage is the primary cause of the monocarpic senescence in wheat.     

Mechanism of monocarpic senescence in rice

During grain formation stage (90 to 110 days), the youngest flag leaf of rice (Oryza sativa L. cv. Jaya) remained metabolically most active (as indicated by cellular constituents and enzyme activities) and the third leaf the least active. At the grain development stage (110 to 120 days) the above pattern of age-related senescence of the flag leaf completely changed and it senesced at a faster rate than the second leaf which remained metabolically active even up to grain maturation time (120 to 130 days), when both the flag and the third leaf partially senesced. Removal of any leaf temporarily arrested senescence of the remaining attached leaves, that of flag leaf did not hasten senescence of the second leaf, while that of either the second or the third accelerated senescence of the flag. Removal of the inflorescence after emergence or foliar treatment of intact plant with kinetin equally delayed senescence and produced an age-related, sequential mode of senescence or leaves. Both translocation and retention of ³²P by the flag leaf were maximum at the time of grain formation and that by the second leaf was maintained even up to grain maturation time. The induction of senescence of the flag leaf was preceded by a plentiful transport of ³²P to the grains. Kinetin treatment decreased the transport of ³²P, prolonged its duration, and almost equally involved all of the leaves in this process. The pattern of senescence of isolated leaf tips was similar to that of attached leaves. The level of endogenous abscisic acid-like substance(s) maintained a close linearity with the senescence behavior of the leaves of intact and defruited plants during aging, and the rise in abscisic acid in the flag leaf was also preceded by higher ³²P transport to the grains.     

Comparison of phosphorus mobilization during monocarpic senescence in rice cultivars with sequential and non-sequential leaf senescence

The senescence pattern of the three uppermost leaves of four rice (Oryza sativa L.) cultivars viz. Ratna, Jaya, Masuri and Kalojira was analysed in terms of decline of chlorophyll and by measuring [³²P]-phosphate retention and export from leaf to grains during the reproductive development. With the advancement of reproductive development, the cultivars Masuri and Kalojira showed a sequential mode of senescence, but the cultivars Ratna and Jaya showed a non-sequential mode of leaf senescence where the flag leaf senesced earlier than the older second leaf. Foliar spraying with benzyladenine (0.5 mM) significantly delayed, and abscisic acid (0.1 mM) accelerated, leaf senescence. In untreated control plants, the second leaf had the highest export of labelled phosphate among the leaves at the grain formation stage (0–7 days) in Masuri and Kalojira. This was compensated by the flag leaf at the grain development stage (7–14 days), whereas export of [³²P]-phosphate was highest from the flag leaf of Ratna and Jaya at the grain development stage. Compared with the control, benzyladenine treatment caused higher retention of [³²P]-phosphate in the leaves and also export to the grains, but abscisic acid treatment gave lower retention and export of [³²P]-phosphate to the grains. The amount of [³²P]-phosphate export from a mother to a daughter shoot developed in the axil of the second leaf of plants with the panicle removed, was less than that to panicles remaining on control plants of all cultivars. When the panicle had been excised, the greatest export of [³²P]-phosphate took place from the second leaf to the daughter shoot in all cultivars. Excision of the panicle delayed leaf senescence as compared with intact controls and maintained an age-related leaf senescence pattern in all the four cultivars. The results presented here demonstrate that mobilization of phosphorus from leaf to grains, regardless of cultivar or age and position of the leaf, correlates well with the senescence of that leaf.     

晚播小麦叶片衰老代谢和粒重变化的比较研究

对７个小麦品种在晚播条件下的叶片衰老生理特性和粒重变化进行了比较研究。根据小麦叶片的衰老特征相差差异,将７个小麦品种区分为３个类型;后健型、早衰型和中间型。在小麦旗叶的衰老过程中,后健型小麦品种旗叶叶绿素和类胡萝卜素含量显著高于早衰型,脂质过氧化产物ＭＤＡ含量显著低于早衰型,小麦粒重降幅依次为早衰型〉中间型〉后健型,并讨论了活性氧代谢在小麦叶片衰老过程中可能作用。     

Sequential and non-sequential pattern of monocarpic senescence in two rice cultivars

Two rice ( Oryza sativa L.) cultivars viz. Ratna (dwarf, photoperiod insensitive) and Masuri (tall, photoperiod sensitive) were selected to analyse their mode of senescence. At the vegetative stage, leaf senescence, expressed as the loss of chlorophyll and protein and a decline in the activities of catalase and alkaline pyrophosphatase, was found to be a function of chronological age (sequential) in both cultivars. With advancing reproductive development, cultivar Masuri retained this sequential mode but cultivar Ratna showed a non-sequential mode of senescence where the flag leaf senesced earlier than the older second leaf, unlike that observed at the vegetative stage. Masuri showed a more rapid senescence than Ratna. In both cultivars, excision of any leaf during anthesis initially retarded the senescence of the remaining leaves on the defoliated plants but soon after, at the grain maturation stage, the leaf senescence started at a higher rate compared with that of the intact control plant. In Ratna, when either the second or the third leaf was removed, the flag leaf senesced faster than that of the unexcised control plant. In Masuri, when either the flag or the third leaf was removed, the second leaf senesced earlier than that of the intact control. In both cultivars, excision of the third leaf showed the least detrimental effect on yield. The greatest detrimental effect on grain yield per plant was observed in Ratna when the flag leaf was removed and in Masuri when the second leaf was removed. Mobilization of metabolites from the source leaf to the sink and the consequent depletion in the leaf as the cause of senescence is discussed.     

Transgenic rice with low sucrose-phosphate synthase activities retain more soluble protein and chlorophyll during flag leaf senescence

We investigated whether changes in sucrose-phosphate synthase (EC 2.4.1.14, SPS) activity could alter N remobilization during leaf senescence. Transgenic rice (Oryza sativa L. cv. Nipponbare) with low SPS activities and wild-type rice plants were grown with basal N (1.0 mM NH₄NO₃) until the late vegetative stage. Subsequently, half of the plants were transferred to a low N (0.1 mM NH₄NO₃) condition to accelerate leaf senescence, and the others were continuously grown with basal N. With low N supply, the amounts of chlorophyll and soluble protein in flag leaf blades decreased after anthesis in both the low SPS plants and wild-type plants, although the decrease was less in the low SPS plants. Panicle weights were significantly lower in the low SPS plant than in the wild-type plant. These results suggest that the remobilization of N from flag leaves was diminished by suppressing the development of reproductive sinks in the low SPS plant.     

Changes in the Chlorophyll Content and Cytokinin Levels in the Top Three Leaves of New Plant Type Rice During Grain Filling

This paper reports the ways that the differences in leaf senescence are related to grain filling, grain yield, and the dynamics of cytokinins (CKs) in the top three leaves of four field-grown new plant type (NPT) rice, a tropical japonica developed at the International Rice Research Institute, Philippines, to increase the yield potential of rice. The chlorophyll content in leaves decreased from flowering to maturity in all the NPT lines, whereas the grain filling percentage was higher in the fast-senescing NPT line than in slow-senescing NPT line. Grain yield was positively correlated with senescence in the flag leaf. Rapid changes in the CK levels were recorded in the leaves of the fast-senescing line, whereas the CK levels were relatively stable in leaves of the slow-senescing line, suggesting that the dynamics of CKs in the fast-senescing line are vital for fast senescence. There were no significant changes in bioactive CKs, CK O-glucosides (storage CKs), and cis-zeatin derivatives in different leaves of the slow-senescing NPT line between 0 and 3 weeks after flowering, suggesting that the content of these CKs is relatively stable during grain filling. A progressive increase in levels of bioactive CKs was positively correlated with gradual accumulation of CK N-glucosides (inactive CKs) in the top three leaves of the slow-senescing NPT line, whereas the decrease of bioactive CKs in the flag leaf of the fast-senescing line was accompanied by accumulation of CK O-glucosides. These results suggest that there is a higher rate of biosynthesis and/or import of bioactive CKs as well as their turnover which may favor delay of leaf senescence in the slow-senescing NPT line.     

Exogenous Jasmonic Acid and Cytokinin Antagonistically Regulate Rice Flag Leaf Senescence by Mediating Chlorophyll Degradation,Membrane Deterioration,and Senescence-Associated Genes Expression

Although it is well known that jasmonic acid (JA) and cytokinin (CK) are involved in regulating leaf senescence, the antagonistic mechanisms of JA and CK on leaf senescence are still unknown. To explore the antagonistic effects of JA and CK on leaf senescence, we treated detached rice flag leaves with JA and CK under dark conditions, and evaluated their chlorophyll contents, membrane deterioration, and expression levels of chlorophyll-degradation-related genes (CDRGs) and senescence-associated genes (SAGs). Our results demonstrated that exogenous application of JA promoted chlorophyll degradation by enhancing the expression levels of CDRGs, promoted membrane deterioration by accelerating the increases in lipid peroxidation and membrane permeability, enhanced the expression levels of SAGs, and consequently accelerated rice flag leaf senescence. On the other hand, exogenous application of CK retarded chlorophyll degradation by down-regulating the expression levels of CDRGs, retarded membrane deterioration by retarding the increases in lipid peroxidation and membrane permeability, down-regulated the expression levels of SAGs, and consequently delayed rice flag leaf senescence. Furthermore, the senescence-accelerating effect of a certain concentration of JA was nullified by the senescence-retarding effect of a certain concentration of CK. These results suggested that exogenous applications of JA and CK were able to antagonistically regulate flag leaf senescence by mediating chlorophyll degradation, membrane deterioration, and SAGs expression. In addition, our results suggested that the progression of flag leaf senescence might not only depend on the level of JA or CK but also depend on the balance between JA and CK.     

黑暗诱导衰老对不同年代冬小麦品种旗叶光系统Ⅱ功能的影响

小麦品种的更新换代是小麦产量不断提高的重要因素,阐明小麦品种演替过程中不同生理特性的变化对新品种选育具有重要参考价值.旗叶衰老速率快慢是影响小麦产量水平的关键因素,目前对于不同小麦品种衰老过程中旗叶光系统Ⅱ功能的变化规律尚不清楚.本试验选用1941-2014年间河南地区不同时期种植的31个品种,通过黑暗诱导离体叶片衰老,测定旗叶叶绿素荧光诱导动力学参数、叶绿素相对含量的变化,分析了光系统Ⅱ功能的变化规律.结果表明:品种演替过程中旗叶的叶绿素含量逐渐提高,衰老过程中近代品种叶绿素的降解速率低于较早年代品种;旗叶衰老过程中,近代品种荧光诱导动力学曲线的J点上升幅度小于I点;品种更替过程中光系统Ⅱ最大光化学效率和单位面积有活性反应中心数目逐渐增加,但是近代品种降低速率低于较早年代品种.叶绿素含量的变化与未衰老叶片中F_v/F_m没有显著相关性,但是随着衰老程度增加,相关性逐渐增大,且趋势线斜率逐渐提高;光系统Ⅱ单位面积有活性反应中心数目与品种育成时间呈显著正相关,且随着衰老程度增加,相关程度和趋势线斜率均显著提高.综上,小麦品种演替过程中,旗叶叶绿素含量逐渐升高,降解速率逐渐减缓,光合电子传递过程中Q_A到Q_B电子传递的抗衰老能力得到改善,从而减缓了衰老过程中光系统Ⅱ最大光化学效率和有活性反应中心的衰减速率,同时,叶绿素含量的提高和旗叶光系统Ⅱ抗衰老能力的增强也是品种更替过程中产量逐渐提高的重要因素.     

Regulation by kinetin and abcisic acid of correlative senescence in relation to grain maturation,source-sink relationship and yield of rice (Oryza sativa L.)

When kinetin was applied to the source organ (flag leaf) of rice (Oryza sativa L. cv. Ratna), foliar senescence was delayed and grain yield per plant (as evidenced by grain weight, grain/straw weight ratio and 1,000 grain growth) was increased through the increase of sink activity (increase in dry weight of the grains/plant), duration of sink capacity as well as photosynthetic ability of the glumes (as determined by the chlorophyll content of the glumes of the developing grains). However, application of kinetin to the sink organs (fruits), promoted senescence of the source but increased the yield by increasing the sink capacity and 1,000 grain growth mostly at the earlier stage of reproductive development. Lower sterility percentage was associated with higher grain yield of the plant by kinetin treatments. ABA applied either to the source or the sink promoted leaf senescence and reduced the grain yield by reducing the sink activity, harvest index, sink capacity duration and increasing the sterility percentage. Thousand grain dry weight at harvest did not vary significantly amongst the treatments. It was concluded that nutrient drainage was associated with the correlative influence of fruit on the monocarpic senescence of rice plant and that a competetion for differential allocation of cytokinin and ABA in the source and sink organs initiates this senescence syndrome.     

Differences in Sugar Accumulation and Mobilization between Sequential and Non-Sequential Senescence Wheat Cultivars under Natural and Drought Conditions

Wheat leaf non-sequential senescence at the late grain-filling stage involves the early senescence of younger flag leaves compared to that observed in older second leaves. On the other hand, sequential senescence involves leaf senescence that follows an age-related pattern, in which flag leaves are the latest to undergo senescence. The characteristics of sugar metabolism in two sequential senescence cultivars and two non-sequential senescence cultivars under both natural and drought conditions were studied to elucidate the underlying mechanism of drought tolerance in two different senescence modes. The results showed that compared to sequential senescence wheat cultivars, under natural and drought conditions, non-sequential senescence wheat cultivars showed a higher leaf net photosynthetic rate, higher soluble sugar levels in leaves, leaf sheaths, and internodes, higher leaf sucrose synthase (SS) and sucrose phosphate synthase (SPS) activity, and higher grain SS activity, thereby suggesting that non-sequential senescence wheat cultivars had stronger source activity. Spike weight, grain weight per spike, and 100-grain weight of non-sequential senescence cultivars at maturity were significantly higher than those of sequential senescence cultivars under both natural and drought conditions. These findings indicate that the higher rate of accumulation and the higher mobilization of soluble sugar in the leaves, leaf sheaths and internodes of non-sequential senescence cultivars improve grain weight and drought tolerance. At the late grain-filling stage, drought conditions adversely affected leaf chlorophyll content, net photosynthetic rate, soluble sugar and sucrose content, SS and SPS activity, gain SS activity, and weight. This study showed that higher rates of soluble sugar accumulation in the source was one of the reasons of triggering leaf non-sequential senescence, and higher rates of soluble sugar mobilization during leaf non-sequential senescence promoted high and stable wheat yield and drought tolerance.     

冷型小麦旗叶衰老和活性氧代谢特性研究

以典型的冷型小麦和暖型小麦为试验材料,研究了同一环境背景下不同温度型小麦开花后的旗叶衰老和活性氧代谢特性。结果表明,与暖型小麦相比,冷型小麦籽料灌浆期旗叶叶绿素和可溶性蛋白质含量下降缓慢、含量高,整个业粒形成和灌浆期ＭＤＡ积累速度慢、含量低,籽粒灌浆期防御活性氧伤害的关键性保护酶（ＳＯＤ、ＣＡＴ和ＰＯＤ）活性下隆幅度小,灌浆中后期活性水平高。由此认为,小麦旗叶衰老和活性氧代谢特性与其温度型的归属关     

麦秸翻压还田对隔茬冬小麦旗叶抗性的生理效应

为了揭示秸秆翻压还田对冬小麦旗叶叶片衰老及抗性指标的变化规律,试验设置4个小麦秸秆翻压还田量(0、4500、6000、7500kg/hm2),测定分析了冬小麦旗叶叶绿素含量、光合速率、丙二醛(MDA)含量、超氧化物歧化酶(SOD)活性、过氧化物酶(POD)活性及可溶性蛋白质含量等抗性指标。结果表明,秸秆还田可明显减缓冬小麦植株衰老过程中叶片叶绿素的降解和光合速率下降,并有效调节叶片超氧化物歧化酶(SOD)、过氧化物酶(POD)活性下降、可溶性蛋白质含量下降和丙二醛(MDA)含量的增加,延缓了冬小麦生育后期叶片的衰老;通过对各抗性指标与光合速率的相关性分析表明,各抗性指标在抑制冬小麦衰老过程中的作用大小依次为:丙二醛>可溶性蛋白质>POD活性>SOD的活性(丙二醛-0.999**;可溶性蛋白质0.997**;POD活性0.976*;SOD活性0.954*)。综合各处理的抗性指标得出,处理Ⅲ(6000kg/hm2)在提高作物后期抗逆性能力方面表现较好,处理Ⅳ(7500kg/hm2)表现较差。     

Comparison of the Drought Stress Responses of Tolerant and Sensitive Wheat Cultivars During Grain Filling: Changes in Flag Leaf Photosynthetic Activity,ABA Levels,and Grain Yield

Water status parameters, flag leaf photosynthetic activity, abscisic acid (ABA) levels, grain yield, and storage protein contents were investigated in two drought-tolerant (Triticum aestivum L. cv. MV Emese and cv. Plainsman V) and two drought-sensitive (cvs. GK élet and Cappelle Desprez) wheat genotypes subjected to soil water deficit during grain filling to characterize physiological traits related to yield. The leaf water potential decreased earlier and at a higher rate in the sensitive than in the tolerant cultivars. The net CO₂ assimilation rate (P _N) in flag leaves during water deficit did not display a strict correlation with the drought sensitivity of the genotypes. The photosynthetic activity terminated earliest in the tolerant cv. Emese, and the senescence of flag leaves lasted 7 days longer in the sensitive Cappelle Desprez. Soil drought did not induce characteristic differences between sensitive and tolerant cultivars in chlorophyll a fluorescence parameters of flag leaves during post-anthesis. Changes in the effective quantum yield of PSII (Φ_PSII) and the photochemical quenching (qP) depended on the genotypes and not on the sensitivity of cultivars. In contrast, the levels of ABA in the kernels displayed typical fluctuations in the tolerant and in the sensitive cultivars. Tolerant genotypes exhibited an early maximum in the grain ABA content during drought and the sensitive cultivars maintained high ABA levels in the later stages of grain filling. In contrast with other genotypes, the grain number per ear did not decrease in Plainsman and the gliadin/glutenin ratio was higher than in the control in Emese during drought stress. A possible causal relationship between high ABA levels in the kernels during late stages of grain filling and a decreased grain yield was found in the sensitive cultivars during drought stress.     

Quantitative proteomics analysis of proteins involved in leaf senescence of rice (<Emphasis Type="Italic">Oryza sativa</Emphasis> L.)

Delaying leaf senescence and prolonging the available time for photosynthesis is one of the important approaches to increase grain yield of rice (Oryza sativa L.). Here, iTRAQ-based quantitative proteomics approach was used to comparative analyze the expression profiles of proteins in rice leaves in response to senescence. Totally 5067 proteins were identified. Compared with the proteins in the flag leaves at early stage of grain-filling in rice Liang-You-Pei 9 (LYP9), 240 and 188 proteins were up-regulated and down-regulated in the flag leaves at middle stage of grain-filling, and 387 and 202 proteins were up-regulated and down-regulated in the flag leaves at late stage of grain-filling, respectively. In addition, 39 and 18 identified proteins were constantly up-regulated and down-regulated in the leaves from early to middle and late stages of grain-filling, respectively. Among them, chloroplast chaperonin 10, geranylgeranyl diphosphate reductase, Mg chelatase subunit ChLD, porphobilinogen deaminase, protochlorophyllide reductase B and thioredoxin-like protein CITRX might have involved in the senescence of leaves. This study provided important information for understanding the age-sensitive mechanism of LYP9, and offered a foundation for future studying and improving it.     

糜子根系与旗叶协同衰老的生理生化机理研究

以榆糜3号糜子为试验材料,测定了拔节至成熟期糜子根系与旗叶生长指标、保护酶活性、可溶性蛋白含量等变化,初步分析了其根系与旗叶协同衰老的生理生化机制.结果表明:从拔节期到成熟期,糜子根系活性、表面积和干物质重,旗叶绿叶面积和干物质重,以及根系与旗叶的超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)活性均呈先升高后降低的趋势,并均在拔节后34~41 d(籽粒灌浆前期~中期)达到最大值;根系与旗叶的可溶性蛋白质含量逐渐降低,而丙二醛(MDA)含量却持续大幅度上升;糜子旗叶的保护酶(SOD、CAT、POD)活性、可溶性蛋白含量始终极显著高于同期根系,而其MDA含量却始终极显著低于同期根系.研究发现,在糜子生育后期,其光合产物分配中心转移,造成根系和旗叶中光合产物亏缺;同时其旗叶和根系中保护酶活性逐渐降低,清除活性氧的能力减弱,引起细胞内活性氧积累和膜氧化伤害逐渐加重,最终导致植株逐渐衰老;在植株衰老过程中,根系与叶片衰老密切相关,并且根系抗氧化酶的活性相对较弱.     

Effect of spikelet removal on the whole plant senescence of rice

The rice (Oryza sativa L.) cultivar IET 1444 showed a nonsequential mode of senescence as evident from the decline in chlorophyll and protein of the flag and second leaves at the senescent stage. Removal of 50,75 and 100 % spikelets from the panicle of rice plant or emasculation of the panicle by hot water treatment induced the development of secondary branch from the axil of second leaf but 25 % removal had no effect. Similarly, removal of 75 and 100 per cent spikelets from the panicle of secondary branch induced tertiary branch development, while 25 and 50 per cent removal had no such effect. Similar treatments on tertiary branch had no effect on further branch production. The pattern of leaf senescence of the untreated (control) main tiller, secondary and tertiary branches was identical, i. e. nonsequential, which could be changed into the sequential type only by the development of additional sinks (i. e. side branch). The leaf area and the seed number of secondary and tertiary branches were gradually reduced and reached a critical value in the tertiary branch. The removal of spikelets or emasculation delayed leaf senescence of the main tiller and the secondary and tertiary branches. Also the longevity of the whole plant could be increased by 40 d i. e., up to the senescence of the tertiary branch. Both leaves and reproductive parts control side branch production, which, in turn, controls the longevity of the whole rice plant.     

Physiological characterization of 'stay green' mutants in durum wheat

Four mutants with delayed leaf senescence were selected from seed of durum wheat mutagenized with ethylmethane sulphonate. Changes in net photosynthetic rate, efficiency of photosystem II and chlorophyll concentration during the maturation and senescence of the flag leaves of both mutant and parental plants were determined under glasshouse conditions. The four mutant lines maintained photosynthetic competence for longer than the parental line and are therefore functionally 'stay green'. The mutant lines also had higher seed weights and grain yields per plant than the parental line.