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Heavy losses by dark leaf spot disease in oilseed Brassica have incited research towards identifying sources of genetic tolerance against causal pathogen, Alternaria brassicicola . Several morpho-molecular parameters were evaluated to test the performance of field mustard and rapeseed genotypes under artificial inoculation with this pathogen. During Brassica - Alternaria interaction, physio-biochemical defense response was witnessed in tolerant genotypes. Two tolerant genotypes (one for field mustard and one for rapeseed), i.e., EC250407 and EC1494 were identified. However, necrotic lesions were more prominent in susceptible genotypes with minimum chlorophyll (chlorophyll a , chlorophyll b and total chlorophyll) and carotenoids contents. Contrary to photosynthetic pigments, increase in total soluble protein (TSP) contents was observed with disease progression in susceptible genotypes. Tolerant genotypes of field mustard and rapeseed displayed remarkable increase in the activities of redox enzyme in infected leaves with least yield loss (6.47% and 5.74%) and disease severity index (DSI) of 2.9 and 2.1, respectively. However, yield/plant showed close association with other morpho-yield parameters, photosynthetic pigments and redox enzymes (superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD)) activities except silique length and TSP. Based on the results of morpho-biochemical analyses, redox enzymes and morphological parameters; their interplay is proposed to determine the tolerance outcome of the Brassica - A . brassicicola interaction.

We altered the chlorophyll (Chl) binding sites in various versions of water-soluble chlorophyll protein (WSCP) by amino acid exchanges to alter their preferences for either Chl a or Chl b . WSCP is ideally suited for this mutational analysis since it forms a tetrameric complex with only four identical Chl binding sites. A loop of 4–6 amino acids is responsible for Chl a versus Chl b selectivity. We show that a single amino acid exchange within this loop changes the relative Chl a / b affinities by a factor of 40. We obtained crystal structures of this WSCP variant binding either Chl a or Chl b . The Chl binding sites in these structures were compared with those in the major light-harvesting complex (LHCII) of the photosynthetic apparatus in plants to search for similar structural features involved in Chl a / b binding specificity. Water-soluble chlorophyll protein forms a tetrameric complex with only four identical chlorophyll-binding sites. Mutational and structural analysis identify the basis of selectivity between a and b forms of chlorophyll.

Copper (Cu) is an essential element for most living plants, but it is toxic for plants when present in excess. To better understand the response mechanism under excess Cu in plants, especially in flowers, transcriptome sequencing on petunia buds and opened flowers under excess Cu was performed. Interestingly, the transcript level of FIT-independent Fe deficiency response genes was significantly affected in Cu stressed petals, probably regulated by basic-helix-loop-helix 121 (bHLH121), while no difference was found in Fe content. Notably, the expression level of bHLH121 was significantly down-regulated in petals under excess Cu. In addition, the expression level of genes related to photosystem II (PSII), photosystem I (PSI), cytochrome b6/f complex, the light-harvesting chlorophyll II complex and electron carriers showed disordered expression profiles in petals under excess Cu, thus photosynthesis parameters, including the maximum PSII efficiency (FV/FM), nonphotochemical quenching (NPQ), quantum yield of the PSII (ΦPS(II)) and photochemical quenching coefficient (qP), were reduced in Cu stressed petals. Moreover, the chlorophyll a content was significantly reduced, while the chlorophyll b content was not affected, probably caused by the increased expression of chlorophyllide a oxygenase (CAO). Together, we provide new insight into excess Cu response and the Cu–Fe crosstalk in flowers.

MYB transcription factors (TFs) regulate diverse plant developmental processes and understanding their roles in controlling pigment accumulation in fruit is important for developing new cultivars. In this study, we characterised kiwifruit TF MYB7, which was found to activate the promoter of the kiwifruit lycopene beta-cyclase (AdLCY-β) gene that plays a key role in the carotenoid biosynthetic pathway. To determine the role of MYB7, we analysed gene expression and metabolite profiles in Actinidia fruit which show different pigment profiles. The impact of MYB7 on metabolic biosynthetic pathways was then evaluated by overexpression in Nicotiana benthamiana followed by metabolite and gene expression analysis of the transformants. MYB7 was expressed in fruit that accumulated carotenoid and Chl pigments with high transcript levels associated with both pigments. Constitutive over-expression of MYB7, through transient or stable transformation of N. benthamiana, altered Chl and carotenoid pigment levels. MYB7 overexpression was associated with transcriptional activation of certain key genes involved in carotenoid biosynthesis, Chl biosynthesis, and other processes such as chloroplast and thylakoid membrane organization. Our results suggest that MYB7 plays a role in modulating carotenoid and Chl pigment accumulation in tissues through transcriptional activation of metabolic pathway genes.

Some cyanobacteria are able to use the far-red end of the light spectrum by synthesizing chlorophyll f pigments. Introducing the protein responsible for chlorophyll f synthesis into crop plants could potentially expand the range of wavelengths that such plants use during photosynthesis and thereby increase their growth efficiency. Ho et al. identified chlorophyll f synthase (ChlF) in two cyanobacteria that are acclimatized to grow using far-red light. Introducing the ChlF-encoding gene into a model cyanobacterium allowed the organism to synthesize chlorophyll f. Similarities between ChlF and a core protein of photosystem II suggest that they have a close evolutionary relationship, and ChlF may even represent a more primitive photochemical reaction center. Science , this issue p. 886 An ancestor of photosystem II allows for oxygenic photosynthesis in the far-­red spectral region. Chlorophyll f (Chl f) permits some cyanobacteria to expand the spectral range for photosynthesis by absorbing far-red light. We used reverse genetics and heterologous expression to identify the enzyme for Chl f synthesis. Null mutants of “super-rogue” psbA4 genes, divergent paralogs of psbA genes encoding the D1 core subunit of photosystem II, abolished Chl f synthesis in two cyanobacteria that grow in far-red light. Heterologous expression of the psbA4 gene, which we rename chlF , enables Chl f biosynthesis in Synechococcus sp. PCC 7002. Because the reaction requires light, Chl f synthase is probably a photo-oxidoreductase that employs catalytically useful Chl a molecules, tyrosine Y Z , and plastoquinone (as does photosystem II) but lacks a Mn 4 Ca 1 O 5 cluster. Introduction of Chl f biosynthesis into crop plants could expand their ability to use solar energy.

Aluminum (Al) is one of the most important stress factors that reduce plant productivity in acidic soils. Present work thereby analyzed Al-induced genomic alterations in Bryophyllum daigremontianum clones using RAPD and ISSR markers, and investigated responding changes in photosynthetic pigment (chlorophyll a, b, a/b, total chlorophyll and carotenoid) contents and total soluble protein amounts in plant leaves. The main reason for the use of bulbiferous spurs originated clone plants was to increase reliability and acceptability of RAPD and ISSR techniques in DNA fingerprinting. Raised 40 clone plants were divided into five separate groups each with eight individuals and each experimental group was watered with 0 (control), 0 (acid control), 50, 100 and 200 µM AlCl3-containing Hoagland solutions on alternate days for two and a half months. All plant soils except control group were sprayed with 0.2% sulfuric acid following watering days and this contributed acidic characteristic (pH 4.8) to soil structure. Increase in Al concentrations were accompanied by an increase in total soluble protein amounts, a decrease in photosynthetic pigment contents, and with appearance, disappearance and intensity changes at RAPD and ISSR band profiles. Out of tested RAPD1-25 and ISSR1-15 primers, RAPD8, RAPD9, ISSR2 and ISSR7 primers produced reproducible band profiles that were distinguishable between treatment and control groups. Findings showed that RAPD and ISSR fingerprints have been useful biomarkers for investigation of plant genotoxicity, especially in clone plants. Moreover, if these fingerprints are integrated with other physiological parameters they could become more powerful tools in ecotoxicology.

Nonphotochemical quenching (NPQ) is a proxy for photoprotective thermal dissipation processes that regulate photosynthetic light harvesting. The identification of NPQ mechanisms and their molecular or physiological triggering factors under in vivo conditions is a matter of controversy. Here, to investigate chlorophyll (Chl)–zeaxanthin (Zea) excitation energy transfer (EET) and charge transfer (CT) as possible NPQ mechanisms, we performed transient absorption (TA) spectroscopy on live cells of the microalga Nannochloropsis oceanica. We obtained evidence for the operation of both EET and CT quenching by observing spectral features associated with the Zea S₁ and Zea●+ excited-state absorption (ESA) signals, respectively, after Chl excitation. Knockout mutants for genes encoding either violaxanthin deepoxidase or LHCX1 proteins exhibited strongly inhibited NPQ capabilities and lacked detectable Zea S₁ and Zea●+ ESA signals in vivo, which strongly suggests that the accumulation of Zea and active LHCX1 is essential for both EET and CT quenching in N. oceanica.

PIF3 is a phytochrome-interacting basic helix-loop-helix transcription factor that negatively regulates light responses, including hypocotyl elongation, cotyledon opening, and hypocotyl negative gravitropism. However, the role of PIF3 in chlorophyll biosynthesis has not been clearly defined. Here, we show that PIF3 also negatively regulates chlorophyll biosynthesis by repressing biosynthetic genes in the dark. Consistent with the gene expression patterns, the etiolated pif3 mutant accumulated a higher amount of protochlorophyllide and was bleached severely when transferred into light. The photobleaching phenotype of pif3 could be suppressed by the gun5 mutation and mimicked by overexpression of GUN5. When 4 negative phytochrome-interacting protein genes (PIF1, PIF3, PIF4, and PIF5) were mutated, the resulting quadruple mutant seedlings displayed constitutive photomorphogenic phenotypes, including short hypocotyls, open cotyledons, and disrupted hypocotyl gravitropism in the dark. Microarray analysis further confirmed that the dark-grown quadruple mutant has a gene expression pattern similar to that of red light-grown WT. Together, our data indicate that 4 phytochrome-interacting proteins are required for skotomorphogenesis and phytochromes activate photomorphogenesis by inhibiting these factors.

Chloroplasts of photosynthetic organisms harness light energy and convert it into chemical energy. In several land plants, GOLDEN2-LIKE (GLK) transcription factors are required for chloroplast development, as glk1 glk2 double mutants are pale green and deficient in the formation of the photosynthetic apparatus. We show here that glk1 glk2 double mutants of Arabidopsis thaliana accumulate abnormal levels of chlorophyll precursors and that constitutive GLK gene expression leads to increased accumulation of transcripts for antenna proteins and chlorophyll biosynthetic enzymes. To establish the primary targets of GLK gene action, an inducible expression system was used in combination with transcriptome analysis. Following induction, transcript pools were substantially enriched in genes involved in chlorophyll biosynthesis, light harvesting, and electron transport. Chromatin immunoprecipitation experiments confirmed the direct association of GLK1 protein with target gene promoters, revealing a putative regulatory cis-element. We show that GLK proteins influence photosynthetic gene expression independently of the phyB signaling pathway and that the two GLK genes are differentially responsive to plastid retrograde signals. These results suggest that GLK genes help to coregulate and synchronize the expression of a suite of nuclear photosynthetic genes and thus act to optimize photosynthetic capacity in varying environmental and developmental conditions.

Chloroplasts are extraordinary organelles for photosynthesis and nutrient storage in plants. During leaf senescence or under stress conditions, damaged chloroplasts are degraded and provide nutrients for developing organs. Autophagy is a high-throughput degradation pathway for intracellular material turnover in eukaryotes. Along with chloroplast degradation, chlorophyll, an important component of the photosynthetic machine, is also degraded. However, the chlorophyll degradation pathways under high light intensity and high temperature stress are not well known. Here, we identified and characterized a novel Arabidopsis mutant, sl2 (seedling lethal 2), showing defective chloroplast development and accelerated chlorophyll degradation. Map-based cloning combined with high-throughput sequencing analysis revealed that a 118.6 kb deletion region was associated with the phenotype of the mutant. Complementary experiments confirmed that the loss of function of ATG2 was responsible for accelerating chlorophyll degradation in sl2 mutants. Furthermore, we analyzed chlorophyll degradation under abiotic stress conditions and found that both chloroplast vesiculation and autophagy take part in chlorophyll degradation under high light intensity and high temperature stress. These results enhanced our understanding of chlorophyll degradation under high light intensity and high temperature stress.