Recent overview of the Mg branch of the tetrapyrrole biosynthesis leading to chlorophylls

In plants, chlorophylls (chlorophyll a and chlorophyll b) are the most abundant tetrapyrrole molecules and are essential for photosynthesis. The first committed step of chlorophyll biosynthesis is the insertion of Mg²⁺ into protoporphyrin IX, and thus subsequent steps of the biosynthesis are called the Mg branch. As the Mg branch in higher plants is complex, it was not until the last decade—after many years of intensive research—that most of the genes encoding the enzymes for the pathway were identified. Biochemical and molecular genetic analyses have certainly modified the classic metabolic map of tetrapyrrole biosynthesis, and only recently have the molecular mechanisms of regulatory pathways governing chlorophyll metabolism been elucidated. As a result, novel functions of tetrapyrroles and biosynthetic enzymes have been proposed. In this review, I summarize the recent findings on enzymes involved in the Mg branch, mainly in higher plants.     

Mechanism and Significance of Chlorophyll Breakdown

Chlorophyll breakdown is the most obvious sign of leaf senescence and fruit ripening. A multistep pathway has been elucidated in recent years that can be divided into two major parts. In the first phase, which commonly is active in higher plants, chlorophyll is converted via several photoreactive intermediates to a primary colorless breakdown product within the chloroplast. The second part of chlorophyll breakdown takes place in the cytosol and the vacuole. During this phase, the primary colorless intermediate is modified in largely species-specific reactions to a number of similar, yet structurally different, linear tetrapyrrolic products that finally are stored within the vacuole of senescing cells. To date, most of the biochemical reactions of the first phase of chlorophyll breakdown have been elucidated and genes have been identified. By contrast, mechanisms of catabolite transport and modification during the second phase are largely unknown. This review summarizes the current knowledge on the biochemical reactions involved in chlorophyll breakdown, with a special focus on the second-phase reactions and the fate of by-products that are released from chlorophyll during its breakdown.     

Toxicity of Fusarium mycotoxins on maize plants

Synergistic effects of Fusarium — toxins mixture (moniliformin, fumonisin B₁, fusaproliferin, zearalenone, zearalenol and deoxynivalenol, each at concentration 3.5 μg mL-1) on maize plants of resistant and susceptible cultivars were studied. After 72-hour treatment the biomass production with both cultivars was approximately 6% lower than in the respective controls. In the resistant cultivar chlorophyll content was increased comparing to control, with higher increase in chlorophyll b. In susceptible cultivar chlorophyll content was slightly decreased, particularly with chlorophyll b.No significant differences between treated and non-treated plants were found in the cell ultrastructure. In susceptible plants the young root cells were more vacuolated and plasmolysis occurred in the cells of outer cortex. In leaves of this cultivar also disorganization of thylakoids in some chloroplasts was observed.     

超表达拟南芥2-烯醛还原酶基因对烟草抗旱性的作用机理分析

为研究是否可以利用2-烯醛还原酶(AER)来清除活性氧下游的醛自由基达到提高植物的抗旱性,以超表达拟南芥AER基因烟草和野生型烟草(SR)为研究材料,利用干旱胁迫处理进行抗旱性分析,测定了干旱胁迫及复水后各个烟草株系的生物量、光合速率、叶绿素荧光参数、叶绿素含量、MDA和H2O2含量等指标。结果显示:(1)干旱胁迫下,转基因烟草株系的生物量、叶绿素含量、净光合速率、PSⅡ最大光化学效率及H2O2的清除能力均显著高于对照;(2)复水之后,烟草植株的各项生理指标都得到一定程度的恢复,而转基因株系相比于野生型恢复迅速,恢复能力更强。研究认为,超表达AER基因可以通过清除活性氧及其下游醛自由基来提高烟草的抗旱能力。     

Effects of neodymium magneto-priming on seed germination and salinity tolerance in tomato

Earth's biosphere is surrounded by magnetic fields that affect all living organisms. A plant's response to magnetic fields is displayed in terms of its seed's vigor, growth, and yield. Examining seed germination in such magnetic fields is the first step in the investigation of how magnetic fields might be used to enhance plant growth and maximize crop performance. In this study, salinity-sensitive Super Strain-B tomato seeds were primed with the northern and southern poles of neodymium magnets of 150, 200, and 250 mT. The magneto-primed seeds showed a significant increase in germination rate and speed, where the orientation of the magnet was identified as being crucial for germination rate and the orientation of seeds towards the magnet was shown to affect the germination speed. The primed plants exhibited enhanced growth characteristics, including longer shoots and roots, larger leaf area, more root hairs, higher water content, and more tolerance to salinity levels, up to 200 mM NaCl. All magneto-primed plants showed a significant decrease in chlorophyll content, continuous chlorophyll fluorescence yield (Ft), and quantum yield (QY). The salinity treatments decreased all chlorophyll parameters in control plants, significantly, but did not lower such parameters in magneto-primed tomatoes. The results of this study illustrate the positive effects of neodymium magnet on the growth and development of tomato plants in terms of their germination, growth, and salinity tolerance, and negatively affected the chlorophyll content in tomato leaves. © 2023 Bioelectromagnetics Society.     

cPPB‐aE is discovered from photosynthetic benthic dinoflagellates

Although chlorophyll degradation pathways in higher plants have been well studied, little is known about the mechanisms of chlorophyll degradation in microalgae. In this article, we report the occurrence of a chlorophyll a derivative that has never been discovered in photosynthetic organisms. This chlorophyll derivative emits no fluorescence and has a peculiar absorbance peak at 425, 451, 625, and 685 nm. From these features, it was identified as 13²,17³‐cyclopheophorbide a enol (cPPB‐aE), reported as a degradation product of chlorophyll a derived from prey algal cells in heterotrophic protists. We discovered cPPB‐aE in six benthic photosynthetic dinoflagellates that are phylogenetically separated into four clades based on SSU rDNA molecular phylogeny. This is the first report of this chlorophyll derivative in photosynthetic organisms and we suggest that the derivative is used to quench excess light energy.     

Effects of Intermittent and Continuous Light on the Chlorophyll Formation in Etiolated Plants at Various Ages

The effect of the tissue age of dark-grown bean plants on the chlorphyll formation under continuous illumination or short impulses of white light has been studied. It was found that the protochlorophyllide present in the tissue is age-dependent and reaches a plateau at about 10 days of age, as judged by the chlorophyll formed in etiolated plants of various ages after 5 min illumination. The amount of chlorophyll a and chlorophyll b formed under short impulses of while light increases up to about 9 days of age and thereafter decreases. However, the decrease in chlorophyll a is sharper than that of chlorophyll b, the amount of which remains almost constant. The ratio of chlorophyll a lo chlorophyll b under the short impulses of white light is higher in the younger plant. Similar results are obtained after transfer of the plants from the flashing light to continuous illumination In the young plant there is no lag phase in the chlorophyll biosynthesis while as the age is increased the lag phase is evident and its duration increases as the plant ages. After protochlorophyllide phototransformation under continuous illumination the lag phase in chlorophyll biosynthesis is also age-dependent. Leaves up to 5 days old show no lag phase in chlorophyll synthesis; after this point, however, the lag phase's duration increases continuously with age.     

Physiological and Biochemical Changes in Lucerne (Medicago sativa) Plants Infected with ‘Candidatus Phytoplasma australasia’-Related Strain (16SrII-D Subgroup)

Changes in physiological and biochemical patterns in lucerne plants caused by the presence of ‘Candidatus Phytoplasma australasia’, which is one of the significant pathogens causing yield losses in lucerne plants, were investigated. Significant differences were evident in total chlorophyll, chlorophyll a, chlorophyll b, and protein amounts between ‘Ca. Phytoplasma australasia’-positive and negative lucerne plants. Stress-related metabolites such as phenol, malondialdehyde, and proline accumulations in ‘Ca. Phytoplasma australasia’-positive plants were remarkably higher than those of phytoplasma-negative plants. As a response to disease attack, phytoplasma-positive plants exhibited higher antioxidant enzymes (peroxidase and catalase) and non-enzymatic metabolite responses such as jasmonic and salicylic acids. We state that partial disease responses were revealed for the first time to breed resistant lucerne lines infected by ‘Ca. Phytoplasma australasia’.     

Mycorrhizal association between the desert truffle Terfezia boudieri and Helianthemum sessiliflorum alters plant physiology and fitness to arid conditions

The host plant Helianthemum sessiliflorum was inoculated with the mycorrhizal desert truffle Terfezia boudieri Chatin, and the subsequent effects of the ectomycorrhizal relationship on host physiology were determined. Diurnal measurements revealed that mycorrhizal (M) plants had higher rates of photosynthesis (35%), transpiration (18%), and night respiration (49%) than non-mycorrhizal (NM) plants. Consequently, M plants exhibited higher biomass accumulation, higher shoot-to-root ratios, and improved water use efficiency compared to NM plants. Total chlorophyll content was higher in M plants, and the ratio between chlorophyll a to chlorophyll b was altered in M plants. The increase in chlorophyll b content was significantly higher than the increase in chlorophyll a content (2.58- and 1.52-fold, respectively) compared to control. Calculation of the photosynthetic activation energy indicated lower energy requirements for CO₂ assimilation in M plants than in NM plants (48.62 and 61.56 kJ mol⁻¹, respectively). Continuous measurements of CO₂ exchange and transpiration in M plants versus NM plants provided a complete picture of the daily physiological differences brought on by the ectomycorrhizal relationships. The enhanced competence of M plants to withstand the harsh environmental conditions of the desert is discussed in view of the mycorrhizal-derived alterations in host physiology.     

Pigment shuffling in antenna systems achieved by expressing prokaryotic chlorophyllide a oxygenase in Arabidopsis

The organization of pigment molecules in photosystems is strictly determined. The peripheral antennae have both chlorophyll a and b, but the core antennae consist of only chlorophyll a in green plants. Furthermore, according to the recent model obtained from the crystal structure of light-harvesting chlorophyll a/b-protein complexes II (LHCII), individual chlorophyll-binding sites are occupied by either chlorophyll a or chlorophyll b. In this study, we succeeded in altering these pigment organizations by introducing a prokaryotic chlorophyll b synthesis gene (chlorophyllide a oxygenase (CAO)) into Arabidopsis. In these transgenic plants (Prochlirothrix hollandica CAO plants), approximately 40% of chlorophyll a of the core antenna complexes was replaced by chlorophyll b in both photosystems. Chlorophyll a/b ratios of LHCII also decreased from 1.3 to 0.8 in PhCAO plants. Surprisingly, these transgenic plants were capable of photosynthetic growth similar to wild type under low light conditions. These results indicate that chlorophyll organizations are not solely determined by the binding affinities, but they are also controlled by CAO. These data also suggest that strict organizations of chlorophyll molecules are not essential for photosynthesis under low light conditions.     

Effects of arbuscular mycorrhizal fungus on photosynthesis and water status of maize under high temperature stress

The purpose of this study was to investigate the effects of arbuscular mycorrhizal (AM) symbiosis on gas exchange, chlorophyll fluorescence, pigment concentration and water status of maize plants in pot culture under high temperature stress. Zea mays L. genotype Zhengdan 958 were cultivated in soil at 26/22°C for 6 weeks, and later subjected to 25, 35 and 40°C for 1 week. The plants inoculated with the AM fungus Glomus etunicatum were compared with the non-inoculated plants. The results showed that high temperature stress decreased the biomass of the maize plants. AM symbiosis markedly enhanced the net photosynthetic rate, stomatal conductance and transpiration rate in the maize leaves. Compared with the non-mycorrhizal plants, mycorrhizal plants had lower intercellular CO₂ concentration under 40°C stress. The maximal fluorescence, maximum quantum efficiency of PSII photochemistry and potential photochemical efficiency of mycorrhizal plants were significantly higher than corresponding non-mycorrhizal plants under high temperature stress. AM-inoculated plants had higher concentrations of chlorophyll a, chlorophyll b and carotenoid than non-inoculated plants. Furthermore, AM colonization increased water use efficiency, water holding capacity and relative water content. In conclusion, maize roots inoculated with AM fungus may protect the plants against high temperature stress by improving photosynthesis and water status.     

The atypical chlorophyll a/b/c light-harvesting complex of Mantoniella squamata: molecular cloning and sequence analysis

cDNA species encoding precursor polypeptides of the chlorophyll a/b/c light-harvesting complex (LHC) of Mantoniella squamata were cloned and sequenced. The precursor polypeptides have molecular weights of 24.2 kDa and are related to the major chlorophyll a/b polypeptides of higher plants. Southern analysis showed that their genes belong to the nuclear encoded Lhc multigene family; the investigated genes most probably do not contain introns. The chlorophyll a/b/c polypeptides contain two highly conserved regions common to all LHC polypeptides and three hydrophobic α-helices, which span the thylakoid membrane. The first membrane-spanning helix, however, is not detected by predictive methods: its atypical hydrophilic domains may bind the chlorophyll c molecules within the hydrophobic membrane environment. Homology to LHC 11 of higher plants and green algae is specifically evident in the C-terminal region comprising helix III and the preceding stroma-exposed domain. The N-terminal region of 29 amino acids resembles the structure of a transit sequence, which shows only minor similarities to those of LHC II sequences. Strikingly, the mature light-harvesting polypeptides of M. squamata lack an N-terminal domain of 30 amino acids, which, in higher plants, contains the phosphorylation site of LHC 11 and simultaneously mediates membrane stacking. Therefore, the chlorophyll a/b/c polypeptides of M. squamata do not exhibit any light-dependent preference for photosystem I or 11. The lack of this domain also indicates that the attractive forces between stacked thylakoids are weak.     

Polypeptide Composition of Photosynthetic Membranes from Chlamydomonas reinhardi and Anabaena variabilis

Anabaena variabilis, a blue-green alga lacking chlorophyll b, shows an absence of the major 22 and 24 kilodalton polypeptides which are present in the photosynthetic membranes of Chlamydomonas reinhardi and higher plants. These data are consistent with other investigations which have shown that these polypeptides are associated with chlorophyll b in the chloroplasts of higher plants, and indicate the presence of a light harvesting chlorophyll-protein complex in higher plants which contains the chlorophyll b of the photosynthetic membrane.     

Phototolerance of lichens, mosses and higher plants in an alpine environment: analysis of photoreactions

 Adaptation to excessive light is one of the requirements of survival in an alpine environment particularly for poikilohydric organisms which in contrast to the leaves of higher plants tolerate full dehydration. Changes in modulated chlorophyll fluorescence and 820-nm absorption were investigated in the lichens Xanthoria elegans (Link) Th. Fr. and Rhizocarpon geographicum (L.) DC, in the moss Grimmia alpestris Limpr. and the higher plants Geum montanum L., Gentiana lutea L. and Pisum sativum L., all collected at altitudes higher than 2000 m above sea level. In the dehydrated state, chlorophyll fluorescence was very low in the lichens and the moss, but high in the higher plants. It increased on rehydration in the lichens and the moss, but decreased in the higher plants. Light-induced charge separation in photosystem II was indicated by pulse-induced fluorescence increases only in dried leaves, not in the dry moss and dry lichens. Strong illumination caused photodamage in the dried leaves, but not in the dry moss and dry lichens. Light-dependent increases in 820-nm absorption revealed formation of potential quenchers of chlorophyll fluorescence in all dehydrated plants, but energy transfer to quenchers decreased chlorophyll fluorescence only in the moss and the lichens, not in the higher plants. In hydrated systems, coupled cyclic electron transport is suggested to occur concurrently with linear electron transport under strong actinic illumination particularly in the lichens because far more electrons became available after actinic illumination for the reduction of photo-oxidized P700 than were available in the pool of electron carriers between photosystems II and I. In the moss Grimmia, but not in the lichens or in leaves, light-dependent quenching of chlorophyll fluorescence was extensive even under nitrogen, indicating anaerobic thylakoid acidification by persistent cyclic electron transport. In the absence of actinic illumination, acidification by ca. 8% CO₂ in air quenched the initial chlorophyll fluorescence yield F_o only in the hydrated moss and the lichens, not in leaves of the higher plants. Under the same conditions, 8% CO₂ reduced the maximal fluorescence yield F_m strongly in the poikilohydric organisms, but only weakly or not at all in leaves. The data indicate the existence of deactivation pathways which enable poikilohydric organisms to avoid photodamage not only in the hydrated but also in the dehydrated state. In the hydrated state, strong nonphotochemical quenching of chlorophyll fluorescence indicated highly sensitive responses to excess light which facilitated the harmless dissipation of absorbed excitation energy into heat. Protonation-dependent fluorescence quenching by cyclic electron transport, P700 oxidation and, possibly, excitation transfer between the photosystems were effectively combined to produce phototolerance. Received: 10 December 1999 / Accepted: 13 April 2000     

Comparison of chlorophyll and carotenoid concentrations among Russian wheat aphid (Homoptera: Aphididae)-infested wheat isolines

Russian wheat aphid, Diuraphis noxia (Mordvilko), feeding injury on 'Betta' wheat isolines with the Dn1 and Dn2 genes was compared by assessing chlorophyll and carotenoid concentrations, and aphid fecundity. The resistant Betta isolines (i.e., Betta-Dn1 and Betta-Dn2) supported similar numbers of aphids, but had significantly fewer than the susceptible Betta wheat, indicating these lines are resistant to aphid feeding. Diuraphis noxia feeding resulted in different responses in total chlorophyll and carotenoid concentrations among the Betta wheat isolines. The infested Betta-Dn2 plants had higher levels of chlorophylls and carotenoids in comparison with uninfested plants. In contrast, infested Betta-Dn1 plants had the same level of chlorophyll and carotenoid in comparison with uninfested plants. Our data provide essential information on the effect of D. noxia feeding on chlorophyll and carotenoid concentrations for Betta wheat and its isolines with D. noxia-resistant Dn1 and Dn2 genes.     

Influence of arbuscular mycorrhizae on photosynthesis and water status of maize plants under salt stress

The influence of arbuscular mycorrhizal (AM) fungus Glomus mosseae on characteristics of the growth, water status, chlorophyll concentration, gas exchange, and chlorophyll fluorescence of maize plants under salt stress was studied in the greenhouse. Maize plants were grown in sand and soil mixture with five NaCl levels (0, 0.5, 1.0, 1.5, and 2.0 g/kg dry substrate) for 55 days, following 15 days of non-saline pretreatment. Under salt stress, mycorrhizal maize plants had higher dry weight of shoot and root, higher relative chlorophyll content, better water status (decreased water saturation deficit, increased water use efficiency, and relative water content), higher gas exchange capacity (increased photosynthetic rate, stomatal conductance and transpiration rate, and decreased intercellular CO(2) concentration), higher non-photochemistry efficiency [increased non-photochemical quenching values (NPQ)], and higher photochemistry efficiency [increased the maximum quantum yield in the dark-adapted state (Fv/Fm), the maximum quantum yield in the light-adapted sate (Fv'/Fm'), the actual quantum yield in the light-adapted steady state (varphiPSII) and the photochemical quenching values (qP)], compared with non-mycorrhizal maize plants. In addition, AM symbiosis could trigger the regulation of the energy biturcation between photochemical and non-photochemical events reflected in the deexcitation rate constants (kN, kN', kP, and kP'). All the results show that G. mosseae alleviates the deleterious effect of salt stress on plant growth, through improving plant water status, chlorophyll concentration, and photosynthetic capacity, while the influence of AM symbiosis on photosynthetic capacity of maize plants can be indirectly affected by soil salinity and mycorrhizae-mediated enhancement of water status, but not by the mycorrhizae-mediated enhancement of chlorophyll concentration and plant biomass.     

Spectroscopic studies of photosystem II in chlorophyll d-containing Acaryochloris marina

Photosystem II (PSII) electron transfer (ET) in the chlorophyll d-containing cyanobacterium Acaryochloris marina (A. marina) was studied by time-resolved electron paramagnetic resonance (EPR) spectroscopy at room temperature, chlorophyll fluorescence, and low-temperature optical spectroscopy. To maximize the ability to measure PSII ET in the intact cells of this organism, growth conditions were optimized to provide the highest specific O(2) activity and the instrumental parameters for the EPR measurements of tyrosine Z (Y(Z)) reduction were adjusted to give the best signal-to-noise over spectral resolution. Analysis of the Y(Z)(*) reduction kinetics revealed that ET to the oxygen-evolving complex on the donor side of PSII in A. marina is indistinguishable from that in higher plants and other cyanobacteria. Likewise, the charge recombination kinetics between the first plastoquinone acceptor Q(A) and the donor side of PSII monitored by the chlorophyll fluorescence decay on the seconds time scale are not significantly different between A. marina and non-chlorophyll d organisms, while low-temperature optical absorption spectroscopy identified the primary electron acceptor in A. marina as pheophytin a. The results indicate that, if the PSII primary electron donor in A. marina is made up of chlorophyll d instead of chlorophyll a, then there must be very different interactions with the protein environment to account for the ET properties, which are similar to higher plants and other cyanobacteria. Nevertheless, the water oxidation mechanism in A. marina is kinetically unaltered.     

超量表达MrCN基因提高烟草植株对TMV的抗性

利用农杆菌介导的遗传转化法将含有普通烟草Ubi．U4启动子驱动MrCN基因表达的元件导入TMV敏感烟草品种K326中,对筛选鉴定出的T0代转基因植株接种TMV,测定其接种前后不同时期的理化指标,以接种TMV的野生型植株为对照。结果显示,转基因植株和野生型植株接种TMV前叶绿素（Chl）和MDA含量YLSOD、POD和CAT酶活力均无显著差异,但接种TMV后野生型植株Chl含量逐渐降低,转基因植株Chl含量先增后降,在接种TMV后5d时转基因植株叶片Chl含量显著高于野生型植株。接种前期（0-3d）转基因植株MDA含量略低于野生型植株,但差异不显著;但接种后期（3～5d）前者的MDA含量显著低于后者。接种TMV后转基因植株中SOD、POD及CAT酶活性变化幅度较野生型植株高,以CAT的变幅最为显著。另外,Real-timePCR分析结果表明,接种TMV后3d时转基因植株删蹦、PR-1α及NrCN表达量均显著高于野生型植株。以上结果表明,通过转基因技术提高烟草抗病基NNrCN的表达量,能提高防御酶活性及病程相关基因的表达量,从而延缓植株感染TMV的发病时间,增强敏感植株对TMV的抗性。     

Senescence-induced ectopic expression of the A. tumefaciens ipt gene in wheat delays leaf senescence, increases cytokinin content, nitrate influx, and nitrate reductase activity, but does not affect grain yield

The manipulation of cytokinin levels by senescence-regulated expression of the Agrobacterium tumefaciens ipt gene through its control by the Arabidopsis SAG12 (senescence-associated gene 12) promoter is an efficient tool for the prolongation of leaf photosynthetic activity which potentially can affect plant productivity. In the present study, the efficiency of this approach was tested on wheat (Triticum aestivum L.)-a monocarpic plant characterized by a fast switch from vegetative to reproductive growth, and rapid translocation of metabolites from leaves to developing grains after anthesis. When compared with the wild-type (WT) control plants, the SAG12::ipt wheat plants exhibited delayed chlorophyll degradation only when grown under limited nitrogen (N) supply. Ten days after anthesis the content of chlorophyll and bioactive cytokinins of the first (flag) leaf of the transgenic plants was 32% and 65% higher, respectively, than that of the control. There was a progressive increase in nitrate influx and nitrate reductase activity. However, the SAG12::ipt and the WT plants did not show differences in yield-related parameters including number of grains and grain weight. These results suggest that the delay of leaf senescence in wheat also delays the translocation of metabolites from leaves to developing grains, as indicated by higher accumulation of ((15)N-labelled) N in spikes of control compared with transgenic plants prior to anthesis. This delay interferes with the wheat reproductive strategy that is based on a fast programmed translocation of metabolites from the senescing leaves to the reproductive sinks shortly after anthesis.