Leaf life span optimizes annual biomass production rather than plant photosynthetic capacity in an evergreen shrub.

SUMMARY: *Owing to nitrogen (N) translocation towards new leaves, the shedding of old leaves can increase the whole-plant carbon gain. It occurs when their photosynthetic nitrogen use efficiency (PNUE) declines below a given threshold. *Here, we investigated variations in net photosynthetic capacity (A(max)), N resorption and PNUE in populations of Rhododendron ferrugineum presenting different mean leaf life spans (LLS). *Both populations had comparable annual leaf surface area production and A(max) across leaf-age cohorts. Branch photosynthetic capacity was up to 95% higher in the population with the longer LLS mainly because of the high contribution of old leaves to the total leaf area. Despite lower N concentrations, old leaves maintained relatively high A(max) and consequently PNUE that were higher than or similar to the values found in current-year leaves. *As the ratio of PNUE in old to PNUE in new leaves was always higher than the fraction of leaf N resorbed during leaf shedding, we concluded that leaf shedding did not improve plant photosynthetic capacity. We suggest that in R. ferrugineum, leaf shedding is mainly controlled by the leaf storage function and, therefore, that models aiming to explain LLS should not only consider the leaf carbon assimilation function, particularly in nutrient-poor habitats.

Endogenous sink-source interactions and soil nitrogen regulate leaf life-span in an evergreen shrub.

How the balance between exogenous and endogenous nitrogen for shoot growth varies with soil nitrogen availability, and its consequences on leaf life-span, have rarely been studied within a single species in the field. In this study, we investigated two Rhododendron ferrugineum populations with contrasting leaf life-span. Soil nitrogen availability and nitrogen resorption of different leaf age classes were assessed, as were the interactions between plant compartments, using (15)N labelling and sink organ suppression. The population growing on poorer soil had a shorter leaf life-span (17.9 vs 21.5 months) and a higher net contribution of leaf reserves to shoot growth (32% vs 15%), achieved by faster nitrogen resorption and greater shedding of young nitrogen-rich leaves. For both populations, wood contributed to over 40% of shoot nitrogen demand. Both the negative relationship between current-year shoot mass and the percentage of 1-yr-old attached leaves and the delay of leaf shedding after bud removal suggest that shoot development has a strong effect on leaf life-span. Our results suggest that, contrary to the evolutionary response, plastic response to low soil nitrogen could reduce leaf life-span in evergreen plants. In addition, leaf life-span seems to be strongly influenced by the discrepancy between shoot nitrogen demand and soil nitrogen uptake rather than nitrogen demand alone.

Nitrogen resorption and photosynthetic activity over leaf life span in an evergreen shrub, Rhododendron ferrugineum, in a subalpine environment.

Here, the advantages for a shrub of having long vs short-lived leaves was investigated in Rhododendron ferrugineum by following nitrogen(15N) and carbon(14C) resorption and translocation, and photosynthetic capacity over the life span. Mean leaf life span was 19 months. Nitrogen (N) resorption in attached leaves occurred mainly in the first year (23%) and reached a maximum of 31% in the second. Although, resorption was similar in attached and fallen 1-yr-old leaves, it was on average one-third higher in fallen than in attached older leaves. Final N resorption of a leaf compartment reached 41%, half occurring from healthy leaves during the first year. Photosynthetic capacity decreased slightly during the life span. Before shoot growth, plant photosynthesis was mainly supported by 1-yr-old leaves, although the contribution of the 2-yr-old leaves was nonnegligible (15% of the capacity and higher carbon transfer toward the roots). After shoot growth, the current-year leaves made the greatest contribution. Our results suggest that short-lived leaves (half of the cohort) are mainly involved in a photosynthetic function, having a high photosynthetic capacity and drawing most of their resorbed N towards current-year leaves; and long-lived leaves are also involved in a conservative function, increasing N resorption and mean residence time (MRT).

Uncoupling nitrogen requirements for spring growth from root uptake in a young evergreen shrub (Rhododendron ferrugineum).

• Internal cycling of nitrogen (N) was investigated in a subalpine field population of the evergreen shrub Rhododendron ferrugineum during spring growth. • The foliar nitrogen of 5-yr-old-plants was directly labeled with 15 N and subsequently traced to all plant compartments. In addition, 15 N-ammonium uptake was estimated in glasshouse experiments. • Before shoot growth, redistribution of 15 N occurred in the plant without net N transfer. During spring development, the decreases in both leaf 15 N and total N were almost identical in terms of percentage, and most of the 15 N withdrawn from the leaf compartments was recovered in the growing shoots. Net changes in the N contents of the various leaf and woody compartments indicate that internal remobilization (especially from 1-yr-old leaves) could have met most of the N needs of new shoot growth. Simultaneously, the rate of mineral N uptake was very low. • Thus, leaves in young plants provide N for new shoots (by contrast with old individuals) and allow, with woody tissues, almost complete uncoupling of N requirement for spring growth from root uptake.

Wild manihot species do not possess C4 photosynthesis.

Cultivated cassava (Manihot esculenta) has a higher rate of photosynthesis than is usual for C3 plants and photosynthesis is not light saturated. For these reasons it has been suggested that cultivated cassava could be derived from wild species possessing C4 photosynthesis. The natural abundance of 13C and activities of phosphoenolpyruvate carboxylase and phosphoglycolate phosphatase were measured in leaves of 20 wild cassava species to test this hypothesis. All the species studied, including M. flabellifolia the potential wild progenitor of cultivated cassava, clearly exhibited C3 not C4 characteristics.

Characterization of closely related delta-TIP genes encoding aquaporins which are differentially expressed in sunflower roots upon water deprivation through exposure to air.

We isolated five sunflower (Helianthus annuus) cDNAs belonging to the TIP (tonoplast intrinsic protein) family. SunRb7 and Sun gammaTIP (partial sequence) are homologous to tobacco TobRb7 and Arabidopsis gamma-TIP, respectively. SunTIP7, 18 and 20 (SunTIPs) are closely related and homologous to Arabidopsis delta-TIP (SunTIP7 and 20 have already been presented in Sarda et al., Plant J. 12 (1997) 1103-1111). As was previously shown for SunTIP7 and 20, expression of SunTIP18 and SunRb7 in Xenopus oocytes caused an increase in osmotic water permeability demonstrating that they are aquaporins. In roots, in situ hybridization revealed that SunTIP7 and 18 mRNAs accumulate in phloem tissues. The expression of TIP-like genes was studied in roots during 24 h water deprivation through exposure to air. During the course of the treatment, each SunTIP gene displayed an individual response: SunTIP7 transcript abundance increased, SunTIP18 decreased whereas that of SunTIP20 was transitorily enhanced. By contrast, SunRb7 and Sun gammaTIP mRNA levels did not fluctuate. Due to the changes in their transcript levels, it is proposed that SUNTIP aquaporins encoded by delta-TIP-like genes play a role in the sunflower response to drought.

Two TIP-like genes encoding aquaporins are expressed in sunflower guard cells.

SunTIP7 and SunTIP20 are closely related sunflower cDNAs showing a deduced amino acid sequence homologous to proteins of the tonoplast intrinsic protein (TIP) family. Their expression in Xenopus oocytes caused a marked increase in osmotic water permeability (demonstrating that they are water channels) which was sensitive to mercury. In leaves, in situ hybridization revealed that both SunTIP7 and SunTIP20 mRNA accumulated in the guard cells. The possible involvement of SunTIPs in stomatal movement was examined by comparing the time course of transcript accumulation and leaf conductance during the daily cycle and following a water limitation. SunTIP7 mRNA fluctuations fitted changes occurring in leaf conductance. The transcript levels were markedly and systematically increased during stomatal closure. It is suggested that aquaporin SunTIP7 facilitates water exit associated with a decrease in guard cell volume. In the same conditions, the transcript level of SunTIP20 remained constant indicating that SunTIP genes are differentially regulated within the same cell.

Identification and expression of water stress- and abscisic acid-regulated genes in a drought-tolerant sunflower genotype.

We have studied two lines of sunflower (Helianthus annuus L.) selected in the field as drought-tolerant (R1 genotype) or drought-sensitive (S1 genotype). When subjected to drought conditions, the R1 line was able to maintain high leaf water potential longer and wilted later than the S1 line. Therefore, this indicates that R1 tolerance includes a leaf-adaptive response. By subtractive hybridization, we have isolated six different cDNAs (designated sdi for sunflower drought-induced) corresponding to transcripts accumulated in R1 and S1 leaves during adaptive response. Analysis of transcript accumulation in response to drought in both genotypes suggests a preferential expression of three sdi genes in the tolerant line. Abscisic acid-mediated induction, analysed in R1 leaves, was observed for only four sdi genes. Sequence analysis of six sdi clones revealed that five clones were related to known proteins including non specific lipid transfer proteins (nsLTP), early light-induced proteins (ELIP), l-aminocyclopropane-l-carboxylate oxidase (ACC oxidase) or dehydrins, predicted to be involved in a wide range of physiological processes.

Short-term effects of nitrate on sucrose synthesis in wheat leaves.

Experiments were carried out with fully expanded leaves from three-week-old seedlings of wheat (Triticum aestivum L.) raised without NO 3 (sup-) . Nitrate was supplied to the leaves through the transpiration stream in the light. Uptake of NO 3 (sup-) was linear with NO 3 (sup-) concentrations from 0 to 80 mM in the solution. Net sucrose synthesis showed inverse relationships versus nitrate uptake, assimilation, and accumulation, with correlation coefficients close to 1. By contrast, no alteration in sucrose synthesis was observed when KCl was substituted for KNO3 in the uptake solution. Sucrose synthesis was not affected by nitrate in seedlings treated with tungstate which absorbed but did not reduce NO 3 (sup-) . After 10 h, the final amount of sucrose in the tissues was only slightly decreased in the presence of NO 3 (sup-) , indicating that the effect of NO 3 (sup-) did not result from an altered sucrose-storage capacity. Comparison of the carbon skeleton and energy reductant necessary for NO 3 (sup-) and CO2 assimilation is consistent with the hypothesis that the processes of NO 3 (sup-) assimilation and sucrose synthesis compete for photosynthetic energy and carbon.

Adaptation of the Photosynthetic Apparatus in Maize Leaves as a Result of Nitrogen Limitation : Relationships between Electron Transport and Carbon Assimilation.

In maize (Zea mays L., cv Contessa), nitrogen (NO(3) (-)) limitation resulted in a reduction in shoot growth and photosynthetic capacity and in an increase in the leaf zeaxanthin contents. Nitrogen deficiency had only a small effect on the quantum yield of CO(2) assimilation but a large effect on the light-saturated rate of photosynthesis. Linear relationships persisted between the quantum yield of CO(2) assimilation and that of photosystem II photochemistry in all circumstances. At high irradiances, large differences in photochemical quenching and nonphotochemical quenching of Chl a fluorescence as well as the ratio of variable to maximal fluorescence (Fv/Fm) were apparent between nitrogen-deficient plants and nitrogen-replete controls, whereas at low irradiances these parameters were comparable in all plants. Light intensity-dependent increases in nonphotochemical quenching were greatest in nitrogen-deficient plants as were the decreases in Fv/Fm ratio. In nitrogen-deficient plants, photochemical quenching decreased with increasing irradiance but remained higher than in controls at high irradiances. Thermal dissipative processes were enhanced as a result of nitrogen deficiency (nonphotochemical quenching was elevated and Fv/Fm was lowered) allowing PSII to remain relatively oxidised even when carbon metabolism was limited via nitrogen limitation.

Inhibition of the phosphate transporter during isolation of intact chloroplasts from leaves of sunflower.

Chloroplasts isolated from spinach leaves by the mechanical method were intact and exhibited high rates of CO2-dependent oxygen evolution whereas chloroplasts isolated from sunflower leaves by the same technique were also intact but showed only low rates of oxygen evolution. The rate of uptake of orthophosphate (Pi) from the suspending medium with sunflower chloroplasts was less than 20% of that in spinach chloroplasts. The apparent Km for Pi transport was lower in sunflower chloroplasts but uptake was competitively inhibited by 3-phosphoglycerate in chloroplasts from both species. Uptake of malate (via the dicarboxylate transporter) and of ATP (via the adenine nucleotide transporter) was also reduced in sunflower chloroplasts compared to spinach chloroplasts. The endogenous Pi content and total exchangeable phosphate pool of sunflower chloroplasts were less than half that in spinach chloroplasts.Addition of a number of possible protective agents to the grinding medium failed to prevent the loss of photosynthetic activity during mechanical isolation of sunflower chloroplasts. Grinding mixtures of spinach and sunflower leaves together indicated that spinach chloroplasts were not inhibited by the sunflower leaf extract. Chloroplasts isolated from sunflower leaves via protoplasts had high rates of CO2-dependent oxygen evolution. The Vmax and Km for Pi uptake, endogenous Pi content and total exchangeable phosphate pool of chloroplasts isolated from sunflower protoplasts were all similar to spinach chloroplasts. It is concluded that inner envelope membrane proteins are damaged during mechanical isolation of sunflower chloroplasts. The decrease in activity of the phosphate transporter and loss of endogenous phosphate may contribute to the low rates of photosynthesis observed in chloroplasts isolated by the mechanical method from leaves of sunflower and possibly other species.