The expression of early light-inducible proteins (ELIPs) under high-light stress as defense marker in Northern- and Southern European cultivars of barley (Hordeum vulgare)

Clones coding for the two small early light-inducible proteins (ELIP)-gene families of 13.5 and 17 kDa have been used as markers to study the effect of high-light intensities on gene expression in cultivars of barley which were bred for growth in Southern and Northern Europe. The mRNA levels of the light-harvesting chlorophyll a / b protein (LHC-II) and the small subunit of ribulose-1,5-bisphosphate carboxylase (SSU) were determined in addition. These data were correlated to the decay of PSII activity during high-light stress and its recovery. In all cultivars, the induction of ELIP mRNAs by high light was accompanied by a correspondent reduction of the LHC-II mRNA level. Furthermore, the LHC-II mRNA levels observed under low-light conditions used for the growth of the plants, were in all cases found to be inversely related to the amount of the ELIP which could be induced by a high-light treatment. In contrast, the amount of the SSU mRNA was reduced only at the highest investigated light intensity of 2 000 μmol m⁻² s⁻¹. During recovery from light-stress, the activity of PSII was quickly restored in all European cultivars. Of these cultivars, however, the cv. Otis which expressed the highest ELIP levels recovered considerably faster than the cultivar p4266N which accumulated the lowest amounts of ELIP under high light. Thus, it appears likely that ELIPs contribute to the restoration of PSII activity during and after photoinhibition.     

Photoinhibition and photoprotection in senescent leaves of field-grown wheat plants

Photosynthesis, photosystem II (PSII) photochemistry, photoinhibition and the xanthophyll cycle in the senescent flag leaves of wheat (Triticum aestivum L.) plants grown in the field were investigated. Compared to the non-senescent leaves, photosynthetic capacity was significantly reduced in senescent flag leaves. The light intensity at which photosynthesis was saturated also declined significantly. The light response curves of PSII photochemistry indicate that a down-regulation of PSII photochemistry occurred in senescent leaves in particular at high light. The maximal efficiency of PSII photochemistry in senescent flag leaves decreased slightly when measured at predawn but substantially at midday, suggesting that PSII function was largely maintained and photoinhibition occurred in senescent leaves when exposed to high light. At midday, PSII efficiency, photochemical quenching and the efficiency of excitation capture by open PSII centers decreased considerably, while non-photochemical quenching increased significantly. Moreover, compared with the values at early morning, a greater decrease in CO₂ assimilation rate was observed at midday in senescent leaves than in control leaves. The levels of antheraxanthin and zeaxanthin via the de-epoxidation of violaxanthin increased in senescent flag leaves from predawn to midday. An increase in the xanthophyll cycle pigments relative to chlorophyll was observed in senescent flag leaves. The results suggest that the xanthophyll cycle was activated in senescent leaves due to the decrease in CO₂ assimilation capacity and the light intensity for saturation of photosynthesis and that the enhanced formation of antheraxanthin and zeaxanthin at high light may play an important role in the dissipation of excess light energy and help to protect photosynthetic apparatus from photodamage. Our results suggest that the well-known function of the xanthophyll cycle to safely dissipate excess excitation energy is also important for maintaining photosynthetic function during leaf senescence.     

Changes in activity of energy dissipating mechanisms in wheat flag leaves during senescence

Abstract: Excitation energy dissipation, including the xanthophyll cycle, during senescence in wheat flag leaves grown in the field was investigated at midday and in the morning. With progress of senescence, photosynthesis (Pn) and actual PSII photochemical efficiency (ΦPSII) decreased markedly at midday. The decrease in extent of Pn was greater than that of ΦPSII. However, there was no significant decline in Pn and ΦPSII observed in the morning, except in leaves 60 days after anthesis. The kinetics of xanthophyll cycle activity, thermal dissipation (NPQ), and q_f observed at midday during senescence exhibited two distinct phases. The first phase was characterized by an increase of xanthophyll cycle activity, NPQ, and q_f during the first 45 days after anthesis. The second phase took place 45 days after anthesis, characterized by a dramatic decline in the above parameters. However, the qI, observed both at midday and in the morning, always increased along with senescence. A larger proportion of NPQ insensitive to DTT (an inhibitor of the de-epoxidation of V to Z) was also observed in severely senescent leaves. In the morning, only severely senescent leaves showed higher xanthophyll cycle activity, NPQ, q_f, and qI. It was demonstrated that, at the beginning of senescence or under low light, wheat leaves were able to dissipate excess light energy via NPQ, depending on the xanthophyll cycle. However, the xanthophyll cycle was insufficient to protect leaves against photodamage under high light, when leaves became severely senescent. The ratio of (Fj - Fo)/(Fp - Fo) increased gradually during the first 45 days after anthesis, but dramatically increased 45 days after anthesis. We propose that another photoprotection mechanism might exist around reaction centres, activated in severely senescent leaves to protect leaves from photodamage.     

Chlorophyll a fluorescence analysis of high-yield rice (Oryza sativa L.) LYPJ during leaf senescence

Photosystem II (PSII) photochemistry was examined by chlorophyll (Chl) a fluorescence analysis in high-yield rice LYPJ flag leaves during senescence. Parameters deduced from the JIP-test showed that inhibition of the donor side of PSII was greater than that of the acceptor side in hybrid rice LYPJ. The natural senescence process was accompanied by the increased inactivation of oxygen-evolving complex (OEC) and a lower total number of active reaction centers per absorption. It indicated that the inhibition of electron transport caused by natural senescence might be caused partly by uncoupling of the OEC and/or inactivation of PSII reaction centers. Chl fluorescence parameters analyzed in this study suggested that energy dissipation was enhanced in order to protect senescent leaves from photodamage. Nevertheless, considerably reduced PSI electron transport activity was observed at the later senescence. Thus, natural senescence inhibited OEC-PSII electron transport, but also significantly limited the PSII-PSI electron flow.     

Characterization of photosynthetic pigment composition, photosystem II photochemistry and thermal energy dissipation during leaf senescence of wheat plants grown in the field

Photosynthetic pigment composition and photosystem II (PSII) photochemistry were characterized during the flag leaf senescence of wheat plants grown in the field. During leaf senescence, neoxanthin and beta-carotene decreased concomitantly with chlorophyll, whereas lutein and xanthophyll cycle pigments were less affected, leading to increases in lutein/chlorophyll and xanthophyll cycle pigments/chlorophyll ratios. The chlorophyll a/b ratio also increased. With the progression of senescence, the maximal efficiency of PSII photochemistry decreased only slightly in the early morning (low light conditions), but substantially at midday (high light conditions). Actual PSII efficiency, photochemical quenching and the efficiency of excitation capture by open PSII centres decreased significantly both early in the morning and at midday and such decreases were much greater at midday than in the early morning. At the same time, non-photochemical quenching, zeaxanthin and antheraxanthin contents at the expense of violaxanthin increased both early in the morning and at midday, with a greater increase at midday. The results in the present study suggest that a down-regulation of PSII occurred in senescent leaves and that the xanthophyll cycle plays a role in the protection of PSII from photoinhibitory damage in senescent leaves by dissipating excess excitation energy, particularly when exposed to high light.     

追施氮肥时期对冬小麦旗叶叶绿素荧光特性的影响

在大田条件下,研究了不同追氮时期对小麦旗叶叶绿素荧光特性、光合速率及籽粒产量的影响.结果表明,拔节期追肥较起身期或挑旗期追肥,改善了小麦旗叶PSⅡ的活性(F_v/F_o)、光化学最大效率(F_v/F_m)、光化学猝灭系数(qP)、实际量子产量(ΦPSⅡ)及光合速率,降低了籽粒灌浆中前期非辐射能量耗散,有利于叶片所吸收的光能较充分地用于光合作用,提高了籽粒灌浆后期非辐射能量的耗散,减缓了叶片光抑制程度和衰老进程.拔节期追肥可显著增加穗粒数和千粒重,提高产量.     

Characterisation of a cysteine protease cDNA from Lolium multiflorum leaves and its expression during senescence and cytokinin treatment

A cysteine protease cDNA clone (See1) highly homologous to barley aleurain was isolated from Lolium multiflorum leaves. During leaf senescence, expression of the See1 mRNA and protein was strongly enhanced. In dark-incubated leaf segments, cytokinin delayed senescence and reduced expression of both See1 mRNA and protein.     

CO(2) enrichment enhances flag leaf senescence in barley due to greater grain nitrogen sink capacity

Senescence is a highly regulated process which is under genetic control. In monocarpic plants, the onset of fruit development is the most important factor initiating the senescence process. During senescence, a large fraction of plant nutrients is reallocated away from vegetative tissues into generative tissues. Senescence may therefore be regarded as a highly effective salvage mechanism to save nutrients for the offspring. CO(2) enrichment, besides increasing growth and yield of C(3) plants, has often been shown to accelerate leaf senescence. C(3) plants grown under elevated CO(2) experience alterations in their nutrient relations. In particular their tissue nitrogen concentrations are always lower after exposure to elevated CO(2). We used a monocarpic C(3) crop - spring barley (Hordeum vulgare cv. Alexis) - grown in open-top field chambers to test the effects of CO(2) enrichment on growth and yield, on nitrogen acquisition and redistribution, and on the senescence process in flag leaves, at two applications of nitrogen fertilizer. CO(2) enrichment (650 vs. 366 μmol mol(-1)) caused an increase both in biomass and in grain yield by 38% (average of the two fertilizer applications) which was due to increased tillering. Total nitrogen uptake of the crops was not affected by CO(2) treatment but responded solely to the N supply. Nitrogen concentrations in grains and straw were significantly lower (-33 and -24%) in plants grown at elevated CO(2). Phenological development was not altered by CO(2) until anthesis. However, progress of flag leaf senescence as assessed by chlorophyll content, protein content and content of large and small subunit of RubisCO and of cytochrome b559 was enhanced under elevated CO(2) concentrations by approximately 4 days. We postulate that CO(2) enhanced flag leaf senescence in barley crops by increasing the nitrogen sink capacity of the grains.     

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 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.     

The early light-induced protein is also produced during leaf senescence of Nicotiana tabacum

To better understand the genetic controls of leaf senescence, a tobacco (Nicotiana tabacum L. cv. SR1) mRNA that is up-regulated during senescence was isolated by the cDNA-amplified restriction fragment polymorphism method and the cDNA was cloned. The mRNA coded for the early light-induced protein (ELIP), a member of the chlorophyll a/b-binding protein family that has been implicated in assembly or repair of the photosynthetic machinery during early chloroplast development and abiotic stress. A protein antigenically recognized by antibodies to ELIP appeared during senescence with kinetics similar to those of its mRNA. The mRNA, designated ELIP-TOB, was detected earlier when senescence was enhanced by leaf detachment and treatment with 1-amino-cyclopropane-1-carboxylic acid, and was detected later when senescence was retarded by benzyladenine. However, no ELIP-TOB mRNA was seen in the dark even though senescence was accelerated under these conditions. Furthermore, water stress and anaerobiosis stimulated the appearance of ELIP-TOB mRNA before losses of chlorophyll could be detected. We discuss the conditions that may lead to the up-regulation of ELIP-TOB during senescence and speculate as to the role of the gene product in this terminal phase of leaf development. Received: 18 May 2000 / Accepted: 24 June 2000