Barry Osmond

From ecophysiology to phenomics: some implications of photoprotection and shade–sun acclimation in situ for dynamics of thylakoids in vitro

Half a century of research into the physiology and biochemistry of sun–shade acclimation in diverse plants has provided reality checks for contemporary understanding of thylakoid membrane dynamics. This paper reviews recent insights into photosynthetic efficiency and photoprotection from studies of two xanthophyll cycles in old shade leaves from the inner canopy of the tropical trees Inga sapindoides and Persea americana (avocado). It then presents new physiological data from avocado on the time frames of the slow coordinated photosynthetic development of sink leaves in sunlight and on the slow renovation of photosynthetic properties in old leaves during sun to shade and shade to sun acclimation. In so doing, it grapples with issues in vivo that seem relevant to our increasingly sophisticated understanding of ΔpH-dependent, xanthophyll-pigment-stabilized non-photochemical quenching in the antenna of PSII in thylakoid membranes in vitro.

Relative functional and optical absorption cross-sections of PSII and other photosynthetic parameters monitored in situ, at a distance with a time resolution of a few seconds, using a prototype light induced fluorescence transient (LIFT) device

The prototype light-induced fluorescence transient (LIFT) instrument provides continuous, minimally intrusive, high time resolution (~2s) assessment of photosynthetic performance in terrestrial plants from up to 2m. It induces a chlorophyll fluorescence transient by a series of short flashes in a saturation sequence (180 ~1μs flashlets in <380μs) to achieve near-full reduction of the primary acceptor QA, followed by a relaxation sequence (RQA; 90 flashlets at exponentially increasing intervals over ~30ms) to observe kinetics of QA re-oxidation. When fitted by the fast repetition rate (FRR) model (Kolber et al. 1998) the QA flash of LIFT/FRR gives smaller values for FmQA from dark adapted leaves than FmPAM from pulse amplitude modulated (PAM) assays. The ratio FmQA/FmPAM resembles the ratio of fluorescence yield at the J/P phases of the classical O-J-I-P transient and we conclude that the difference simply is due to the levels of PQ pool reduction induced by the two techniques. In a strong PAM-analogous WL pulse in the dark monitored by the QA flash of LIFT/FRR φPSIIWL ≈ φPSIIPAM. The QA flash also tracks PQ pool reduction as well as the associated responses of ETR QA → PQ and PQ → PSI, the relative functional (σPSII) and optical absorption (aPSII) cross-sections of PSII in situ with a time resolution of ~2s as they relax after the pulse. It is impractical to deliver strong WL pulses at a distance in the field but a longer PQ flash from LIFT/FRR also achieves full reduction of PQ pool and delivers φPSIIPQ ≈ φPSIIPAM to obtain PAM-equivalent estimates of ETR and NPQ at a distance. In situ values of σPSII and aPSII from the QA flash with smaller antenna barley (chlorina-f2) and Arabidopsis mutants (asLhcb2-12, ch1-3 Lhcb5) are proportionally similar to those previously reported from in vitro assays. These direct measurements are further validated by changes in antenna size in response to growth irradiance. We illustrate how the QA flash facilitates our understanding of photosynthetic regulation during sun flecks in natural environments at a distance, with a time resolution of a few seconds.

Do daily and seasonal trends in leaf solar induced fluorescence reflect changes in photosynthesis, growth or light exposure?

Solar induced chlorophyll fluorescence (SIF) emissions of photosynthetically active plants retrieved from space-borne observations have been used to improve models of global primary productivity. However, the relationship between SIF and photosynthesis in diurnal and seasonal cycles is still not fully understood, especially at large spatial scales, where direct measurements of photosynthesis are unfeasible. Motivated by up-scaling potential, this study examined the diurnal and seasonal relationship between SIF and photosynthetic parameters measured at the level of individual leaves. We monitored SIF in two plant species, avocado (Persea Americana) and orange jasmine (Murraya paniculatta), throughout 18 diurnal cycles during the Southern Hemisphere spring, summer and autumn, and compared them with simultaneous measurements of photosynthetic yields, and leaf and global irradiances. Results showed that at seasonal time scales SIF is principally correlated with changes in leaf irradiance, electron transport rates (ETR) and constitutive heat dissipation (YNO; p < 0.001). Multiple regression models of correlations between photosynthetic parameters and SIF at diurnal time scales identified leaf irradiance as the principle predictor of SIF (p < 0.001). Previous studies have identified correlations between photosynthetic yields, ETR and SIF at larger spatial scales, where heterogeneous canopy architecture and landscape spatial patterns influence the spectral and photosynthetic measurements. Although this study found a significant correlation between leaf-measured YNO and SIF, future dedicated up-scaling experiments are required to elucidate if these observations are also found at larger spatial scales.

Integrating Transient Heterogeneity of Non-Photochemical Quenching in Shade-Grown Heterobaric Leaves of Avocado (Persea americana L.): Responses to CO2 Concentration, Stomatal Occlusion, Dehydration and Relative Humidity

Long-lived shade leaves of avocado had extremely low rates of photosynthesis. Gas exchange measurements of photosynthesis were of limited use, so we resorted to Chl fluorescence imaging (CFI) and spot measurements to evaluate photosynthetic electron transport rates (ETRs) and non-photochemical quenching (NPQ). Imaging revealed a remarkable transient heterogeneity of NPQ during photosynthetic induction in these hypostomatous, heterobaric leaves, but was adequately integrated by spot measurements, despite long-lasting artifacts from repeated saturating flashes during assays. Major veins (mid-vein, first- and second-order veins) defined areas of more static large-scale heterogeneous NPQ, with more dynamic small-scale heterogeneity most strongly expressed in mesophyll cells between third- and fourth-order veins. Both responded to external CO2 concentration ([CO2]), occlusion of stomata with Vaseline™, leaf dehydration and relative humidity (RH). We interpreted these responses in terms of independent behavior of stomata in adjacent areoles that was largely expressed through CO2-limited photosynthesis. Heterogeneity was most pronounced and prolonged in the absence of net CO2 fixation in 100 p.p.m. [CO2] when respiratory and photorespiratory CO2 cycling constrained the inferred ETR to ~75% of values in 400 or 700 p.p.m. [CO2]. Likewise, sustained higher NPQ under Vaseline™, after dehydration or at low RH, also restricted ETR to ~75% of control values. Low NPQ in chloroplast-containing cells adjacent to major veins but remote from stomata suggested internal sources of high [CO2] in these tissues.

Message from the International Society of Photosynthesis Research (ISPR)

Photosynthesis Congress, held in 2004 in Montreal Canada (PS2004), will receive an ISPR Newsletter, the third in the series after the Montreal meeting, which contains the information presented below. With this message we are trying to reach all scientists interested in photosynthesis, those who are experiencing the fascinating progress occurring in all aspects of this field, from the capture of photons to CO2 fixation and ecosystem studies. Photosynthesis researchers can be proud of the great breakthroughs in structural and functional photosynthesis research. We are becoming increasingly aware that photosynthesis plays an important role in plant growth, development and acclimation, either by regulating the gene expression directly via photosynthesis intermediates and (side) products or by cross signaling with various plant hormones. It is also clear that future energy sources must rely on solar energy conversion and, in this respect, research on biological systems and on the development of biomimetic artificial photosynthesis systems are of utmost importance. These few examples already indicate that research on photosynthesis is now needed more than ever. Our invitation to join ISPR follows.

Experimental ecosystem and climate change research in controlled environments: lessons from the Biosphere 2 Laboratory 1996–2003

It is clear from the project summaries below that the Biosphere 2 Laboratory (B2L) delivered handsomely as a controlled environment facility for experimental ecosystem and global climate change research. Ironically, the short and medium term experiments with model complex systems revealed that some of the most exciting and unexpected questions involved carbon cycling in benthic and soil metabolism, the very same processes that caused the first closed mission in the facility to fail, and eventually made the apparatus available for research. The effects of elevated [CO2] on these processes in the marine and agriforest mesocosms was to stimulate flux and to reduce Csequestration, by reduced carbonate deposition and enhanced metabolism of soil C reserves, respectively. The extent to which this and other themes that emerged from experiments were products of the initial conditions established in B2L model systems, or are general principles that prevail in natural ecosystems, remains to be seen.

Non-intrusive Assessment of Photosystem II and Photosystem I in Whole Coral Tissues

Reef building corals (phylum Cnidaria) harbour endosymbiotic dinoflagellate algae (genus Symbiodinium) that generate photosynthetic products to fuel their host’s metabolism. Non-invasive techniques such as chlorophyll (Chl) fluorescence analyses of Photosystem II (PSII) have been widely used to estimate the photosynthetic performance of Symbiodinium in hospite. However, since the spatial origin of PSII chlorophyll fluorescence in coral tissues is uncertain, such signals give limited information on depth-integrated photosynthetic performance of the whole tissue. In contrast, detection of absorbance changes in the near infrared (NIR) region integrates signals from deeper tissue layers due to weak absorption and multiple scattering of NIR light. While extensively utilised in higher plants, NIR bio-optical techniques are seldom applied to corals. We have developed a non-intrusive measurement method to examine photochemistry of intact corals, based on redox kinetics of the primary electron donor in Photosystem I (P700) and chlorophyll fluorescence kinetics (Fast-Repetition Rate fluorometry, FRRf). Since the redox state of P700 depends on the operation of both PSI and PSII, important information can be obtained on the PSII-PSI intersystem electron transfer kinetics. Under moderate, sub-lethal heat stress treatments (33 ˚C for ~20 min), the coral Pavona decussata exhibited down-regulation of PSII electron transfer kinetics, indicated by slower rates of electron transport from QA to plastoquinone (PQ) pool, and smaller relative size of oxidised PQ with concomitant decrease of a specifically-defined P700 kinetics area, which represents the active pool of PSII. The maximum quantum efficiency of PSII (Fv/Fm) and functional absorption cross-section of PSII (σPSII) remained unchanged. Based on the coordinated response of P700 parameters and PSII-PSI electron transport properties, we propose that simple P700 kinetics parameters as employed here serve as indicators of the integrity of PSII-PSI electron transfer dynamics in corals.