Chlorophyll Synthesis Pathway Research Articles

Integrated physiological, biochemical and transcriptomic analyses reveal the mechanism of salt tolerance induced by a halotolerant Serratia sp. NTN6 in maize

Salt stress is a serious environmental challenge that hinders crop growth and yield. Plant growth-promoting rhizobacteria (PGPR) can successfully and environmentally friendly boost plant development in saline soils. In this study, a halotolerant Serratia sp. NTN6 was isolated from the rhizosphere soil of halophyte Puccinellia tenuiflora and its plant growth-promoting properties were determined. NTN6 inoculation significantly promoted seed germination and seedling growth in maize under salt stress. Shoot height, stem thickness, root fresh weight, shoot fresh weight, root dry weight, and shoot dry weight of salt-stressed seedlings increased by 36.3%, 32.5%, 146.4%, 193.6%, 25.0%, and 109.6%, respectively, after inoculation with NTN6. NTN6-mediated maize salt tolerance was revealed via transcriptomic, physiological, and biochemical analyses. Under salt stress, NTN6 regulated the transcript levels of genes related to chlorophyll synthesis, photosynthesis apparatus, and carbon assimilation pathways, and thus increasing chlorophyll content, Fv/Fm, and net photosynthetic rate; NTN6 reduced the content of O2•–, H2O2, and MDA by regulating the transcripts and activities of antioxidant enzymes; NTN6 affected the transcriptional response of genes involved in ABA synthesis and signaling to salt stress and reduced ABA level. Finally, NTN6 boosted photosynthetic capacity in salt-stressed maize by modulating photochemical activity, C4 pathway and Calvin cycle as well as mitigating reactive oxygen species damage to the photosystem reaction center. This study first investigated the mechanism of Serratia sp.-induced salt tolerance in maize and these findings demonstrate the valuable contribution of halotolerant PGPR strain from halophytes in improving salinity tolerance in non-halophytic crops.

Metabolome and Transcriptome Reveal Chlorophyll, Carotenoid, and Anthocyanin Jointly Regulate the Color Formation of Triadica sebifera.

The Chinese tallow tree (Triadica sebifera) is an economically important plant on account of its ornamental value and oil-producing seeds. Leaf colour is a key characteristic of T. sebifera, with yellow-, red- and purple-leaved varieties providing visually impressive displays during autumn. In this study, we performed metabolomic and transcriptomic analyses to gain a better understanding of the mechanisms underlying leaf colour development in purple-leaved T. sebifera at three stages during the autumnal colour transition, namely, green, hemi-purple, and purple leaves. We accordingly detected 370 flavonoid metabolites and 10 anthocyanins, among the latter of which, cyanidin-3-xyloside and peonidin-3-O-glucoside were identified as the predominant compounds in hemi-purple and purple leaves. Transcriptomic analysis revealed that structural genes associated with the anthocyanin biosynthetic pathway, chlorophyll synthesis pathway and carotenoid synthesis pathway were significantly differential expressed at the three assessed colour stages. Additionally, transcription factors associated with the MYB-bHLH-WD40 complex, including 22 R2R3-MYBs, 79 bHLHs and 44 WD40 genes, were identified as candidate regulators of the anthocyanin biosynthetic pathway. Moreover, on the basis of the identified differentially accumulated anthocyanins and key genes, we generated genetic and metabolic regulatory networks for anthocyanin biosynthesis in T. sebifera. These findings provide comprehensive information on the leaf transcriptome and three pigments of T. sebifera, thereby shedding new light on the mechanisms underlying the autumnal colouring of the leaves of this tree.

MiRNAs are involved in regulating the formation of recovery tissues in virus infected Nicotiana tabacum.

MiRNAs play an important role in regulating plant growth and immune response. Mosaic diseases are recognized as the most important plant diseases in the world, and mosaic symptoms are recovery tissues formed by plants against virus infection. However, the mechanism of the formation of mosaic symptoms remains elusive. In this study, two typical mosaic systems consisting of Nicotiana tabacum-cucumber mosaic virus (CMV) and N. tabacum-tobacco mosaic virus (TMV) were used to investigate the relevance of miRNAs to the appearance of mosaic symptoms. The results of miRNA-seq showed that there were significant differences in miRNA abundance between dark green tissues and chlorotic tissues in mosaic leaves caused by the infection of CMV or TMV. Compared with healthy tissues, miRNA expression was significantly increased in chlorotic tissues, but slightly increased in dark green tissues. Three miRNAs, namely miR1919, miR390a, and miR6157, were identified to be strongly up-regulated in chlorotic tissues of both mosaic systems. Results of overexpressing or silencing of the three miRNAs proved that they were related to chlorophyll synthesis, auxin response, and small GTPase-mediated immunity pathway, which were corresponding to the phenotype, physiological parameters and susceptibility of the chlorotic tissues in mosaic leaves. Besides, the newly identified novel-miRNA48, novel-miRNA96 and novel-miRNA103 may also be involved in this formation of mosaic symptoms. Taken together, our results demonstrated that miR1919, miR390a and miR6157 are involved in the formation of mosaic symptoms and plant antiviral responses, providing new insight into the role of miRNAs in the formation of recovery tissue and plant immunity.

Carbon monoxide is involved in melatonin-enhanced drought resistance in tomato seedlings by enhancing chlorophyll synthesis pathway

BackgroundDrought is thought to be a major abiotic stress that dramatically limits tomato growth and production. As signal molecule, melatonin (MT) and carbon monoxide (CO) can enhance plant stress resistance. However, the effect and underlying mechanism of CO involving MT-mediated drought resistance in seedling growth remains unknown. In this study, tomato (Solanum lycopersicum L. ‘Micro-Tom’) seedlings were used to investigate the interaction and mechanism of MT and CO in response to drought stress.ResultsThe growth of tomato seedlings was inhibited significantly under drought stress. Exogenous MT or CO mitigated the drought-induced impairment in a dose-dependent manner, with the greatest efficiency provided by 100 and 500 µM, respectively. But application of hemoglobin (Hb, a CO scavenger) restrained the positive effects of MT on the growth of tomato seedlings under drought stress. MT and CO treatment promoted chlorophyll a (Chl a) and chlorophyll a (Chl b) accumulations. Under drought stress, the intermediate products of chlorophyll biosynthesis such as protoporphyrin IX (Proto IX), Mg-protoporphyrin IX (Mg-Proto IX), potochlorophyllide (Pchlide) and heme were increased by MT or CO, but uroporphyrinogen III (Uro III) content decreased in MT-treated or CO-treated tomato seedlings. Meanwhile, MT or CO up-regulated the expression of chlorophyll and heme synthetic-related genes SlUROD, SlPPOX, SlMGMT, SlFECH, SlPOR, SlChlS, and SlCAO. However, the effects of MT on chlorophyll biosynthesis were almost reversed by Hb.ConclusionThe results suggested that MT and CO can alleviate drought stress and facilitate the synthesis of Chl and heme in tomato seedlings. CO played an essential role in MT-enhanced drought resistance via facilitating chlorophyll biosynthesis pathway.

Alleviation of Adverse Effects of Drought Stress on Growth and Nitrogen Metabolism in Mungbean (Vigna radiata) by Sulphur and Nitric Oxide Involves Up-Regulation of Antioxidant and Osmolyte Metabolism and Gene Expression.

The influence of drought induced by polyethylene glycol (PEG) and the alleviatory effect of nitric oxide (50 µM) and sulphur (S, 1 mM K2SO4) were studied in Vigna radiata. Drought stress reduced plant height, dry weight, total chlorophylls, carotenoids and the content of nitrogen, phosphorous, potassium and sulphur. The foliar applications of NO and sulphur each individually alleviated the decline, with a greater alleviation observed in seedlings treated with both NO and sulphur. The reduction in intermediates of chlorophyll synthesis pathways and photosynthesis were alleviated by NO and sulphur. Oxidative stress was evident through the increased hydrogen peroxide, superoxide and activity of lipoxygenase and protease which were significantly assuaged by NO, sulphur and NO + sulphur treatments. A reduction in the activity of nitrate reductase, glutamine synthetase and glutamate synthase was mitigated due to the application of NO and the supplementation of sulphur. The endogenous concentration of NO and hydrogen sulphide (HS) was increased due to PEG; however, the PEG-induced increase in NO and HS was lowered due to NO and sulphur. Furthermore, NO and sulphur treatments to PEG-stressed seedlings further enhanced the functioning of the antioxidant system, osmolytes and secondary metabolite accumulation. Activities of γ-glutamyl kinase and phenylalanine ammonia lyase were up-regulated due to NO and S treatments. The treatment of NO and S regulated the expression of the Cu/ZnSOD, POD, CAT, RLP, HSP70 and LEA genes significantly under normal and drought stress. The present study advocates for the beneficial use of NO and sulphur in the mitigation of drought-induced alterations in the metabolism of Vigna radiata.

Scale-down of oxygen and glucose fluctuations in a tubular photobioreactor operated under oxygen-balanced mixotrophy.

Oxygen-balanced mixotrophy (OBM) is a novel type of microalgal cultivation that improves autotrophic productivity while reducing aeration costs and achieving high biomass yields on substrate. The scale-up of this process is not straightforward, as non-ideal mixing in large photobioreactors might have unwanted effects in cell physiology. We simulated at lab scale dissolved oxygen and glucose fluctuations in a tubular photobioreactor operated under OBM where glucose is injected at the beginning of the tubular section. We ran repeated batch experiments with the strain Galdieria sulphuraria ACUF 064 under glucose pulse feeding of different lengths, representing different retention times: 112, 71 and 21 minutes. During the long and medium tube retention time simulations, dissolved oxygen was depleted 15 to 25 minutes after every glucose pulse. These periods of oxygen limitation resulted in the accumulation of coproporphyrin III in the supernatant, an indication of disruption in the chlorophyll synthesis pathway. Accordingly, the absorption cross section of the cultures decreased steeply, going from values of 150 - 180 m2 ·kg-1 at the end of the first batch down to 50 - 70 m2 ·kg-1 in the last batches of both conditions. In the short tube retention time simulation, dissolved oxygen always stayed above 10% air saturation and no pigment reduction nor coproporphyrin III accumulation were observed. Concerning glucose utilization efficiency, glucose pulse feeding caused a reduction of biomass yield on substrate in the range of 4 to 22% compared to the maximum levels previously obtained with continuous glucose feeding (0.9 C-g·C-g-1 ). The missing carbon was excreted to the supernatant as extracellular polymeric substances constituted by carbohydrates and proteins. Overall, the results point out the importance of studying large-scale conditions in a controlled environment and the need for a highly controlled glucose feeding strategy in the scale-up of mixotrophic cultivation. This article is protected by copyright. All rights reserved.

Effects of Cadmium on metabolism of photosynthetic pigment and photosynthetic system in Lactuca sativa L. revealed by physiological and proteomics analysis

Cadmium stress stunted lettuce growth, resulting in phenotypic changes such as leaf yellowing and shrinkage, as well as physiological changes such as reduced chlorophyll and photosynthesis. However, the molecular mechanism by which this metal inhibits lettuce development and plant stress response remains unclear. The effects of 20 μmol/L Cd treatment on chlorophyll and carotenoids contents as well as photosynthetic function of lettuce leaves was investigated to explore the mechanism of action of Cd on photosynthetic function. Using an IBT-based proteomics technique, the key proteins in these physiological processes were quantified. It was found that the expression of important enzymes in the chlorophyll synthesis pathway in leaves, such as HemB, POR and DVR, was shown to be hindered by Cd stress, resulting in a reduction in chlorophyll content. Meanwhile, Cd promoted the degradation of carotenoids and up-regulated the expression of degradation enzymes NCED4 and CCD8, resulting in a reduction in carotenoids content. Furthermore, the expression of the xanthophyll cycle-regulating enzymes ZEP and VDE was dramatically reduced. This means that lettuce's inherent antioxidant defensive mechanisms, such as the xanthophyll cycle and the NPQ-dependent energy dissipation process, are impaired. Moreover, the decrease in chlorophyll and carotenoids content reduced the stability of LHC, where the protein expression of Lhca4, Lhcb1, Lhcb3, Lhcb4 and Lhcb6 was significantly reduced. Additionally, Cd not only reduced the expression of PSII receptor-side proteins (PsbL, PsbP, and PsbR) and core of PSI response center (psaC, psaD, psaE, psaL and psaN), but also electron transport-related proteins (Cyt b6/f complex, Fd and H+-ATPase), carbon fixation-related ATPase subunits and Rubisco subunits expression were all significantly reduced, then the supply of carbon assimilation power was inhibited. Therefore, on one hand, Cd stress inhibited the synthesis of photosynthetic pigments and photosynthesis-related proteins or subunits, eventually affected the fixation of CO2, as manifested by the decreased Pn and plant biomass. On the other hand, Cd stress directly affected the protective mechanism of the photosynthetic apparatus, such as xanthophyll cycle and antioxidant capacity, which ultimately leads to the photodamaged of the photosynthetic apparatus. This study delves into the target sites of Cd attack on lettuce and provides an accurate reference for our research on Cd mitigants.

MbHY5-MbYSL7 mediates chlorophyll synthesis and iron transport under iron deficiency in Malus baccata

Iron (Fe) plays an important role in cellular respiration and catalytic reactions of metalloproteins in plants and animals. Plants maintain iron homeostasis through absorption, translocation, storage, and compartmentalization of iron via a cooperative regulative network. Here, we showed different physiological characteristics in the leaves and roots of Malus baccata under Fe sufficiency and Fe deficiency conditions and propose that MbHY5 (elongated hypocotyl 5), an important transcription factor for its function in photomorphogenesis, participated in Fe deficiency response in both the leaves and roots of M. baccata. The gene co-expression network showed that MbHY5 was involved in the regulation of chlorophyll synthesis and Fe transport pathway under Fe-limiting conditions. Specifically, we found that Fe deficiency induced the expression of MbYSL7 in root, which was positively regulated by MbHY5. Overexpressing or silencing MbYSL7 influenced the expression of MbHY5 in M. baccata.

Integrated metabolomic and transcriptomic analyses reveal different metabolite biosynthesis profiles of Juglans mandshurica in shade.

Light is not only a very important source of energy for the normal growth and development of plants, but also a regulator of many development and metabolic processes. The mechanism of plant growth and development under low light conditions is an important scientific question. With the promulgation of the law to stop natural forest cutting, understory regeneration is an important method for artificial forest afforestation. Here, the growth and physiological indexes of Juglans mandshurica, an important hardwood species in Northeast China, were measured under different shade treatments. In addition, transcriptome and metabolome were compared to analyze the molecular mechanism of shade tolerance in J. mandshurica. The results showed that the seedling height of the shade treatment group was significantly higher than that of the control group, and the 50% light (L50) treatment was the highest. Compared with the control group, the contents of gibberellin, abscisic acid, brassinolide, chlorophyll a, and chlorophyll b in all shade treatments were significantly higher. However, the net photosynthetic rate and water use efficiency decreased with increasing shade. Furthermore, the transcriptome identified thousands of differentially expressed genes in three samples. Using enrichment analysis, we found that most of the differentially expressed genes were enriched in photosynthesis, plant hormone signal transduction and chlorophyll synthesis pathways, and the expression levels of many genes encoding transcription factors were also changed. In addition, analysis of differentially accumulated metabolites showed that a total of 470 differential metabolites were identified, and flavonoids were the major differential metabolites of J. mandshurica under light stress. These results improved our understanding of the molecular mechanism and metabolite accumulation under light stress in J. mandshurica.

The genomic and bulked segregant analysis of Curcuma alismatifolia revealed its diverse bract pigmentation

Compared with most flowers where the showy part comprises specialized leaves (petals) directly subtending the reproductive structures, most Zingiberaceae species produce showy “flowers” through modifications of leaves (bracts) subtending the true flowers throughout an inflorescence. Curcuma alismatifolia, belonging to the Zingiberaceae family, a plant species originating from Southeast Asia, has become increasingly popular in the flower market worldwide because of its varied and esthetically pleasing bracts produced in different cultivars. Here, we present the chromosome-scale genome assembly of C. alismatifolia “Chiang Mai Pink” and explore the underlying mechanisms of bract pigmentation. Comparative genomic analysis revealed C. alismatifolia contains a residual signal of whole-genome duplication. Duplicated genes, including pigment-related genes, exhibit functional and structural differentiation resulting in diverse bract colors among C. alismatifolia cultivars. In addition, we identified the key genes that produce different colored bracts in C. alismatifolia, such as F3′5'H, DFR, ANS and several transcription factors for anthocyanin synthesis, as well as chlH and CAO in the chlorophyll synthesis pathway by conducting transcriptomic analysis, bulked segregant analysis using both DNA and RNA data, and population genomic analysis. This work provides data for understanding the mechanism of bract pigmentation and will accelerate breeding in developing novel cultivars with richly colored bracts in C. alismatifolia and related species. It is also important to understand the variation in the evolution of the Zingiberaceae family.

WAL3 encoding a PLS-type PPR protein regulates chloroplast development in rice

Chloroplast development is a complex process that is critical for the growth and development of plants. Pentapeptide repeat (PPR) proteins contain large members but only few of them have been characterized in rice. In this study, we identified a new PLS-type protein, WAL3 (Whole Albino Leaf on Chromosome 3), playing important roles in plant chloroplast development. Knockout of WAL3 gene in Nipponbare variety caused abnormal chloroplast development and showed an albino lethal phenotype. Expression analysis showed that WAL3 gene was constitutively expressed with the highest expression in leaves. The WAL3 protein localized in chloroplasts and affected the splicing of multiple group II introns. Transcriptome sequencing showed that WAL3 involved in multiple metabolic pathways including the chlorophyll synthesis and photosynthetic related metabolic pathways. The decreased abundance of photosynthesis-related proteins in wal3 mutants indicated WAL3 influence photosynthesis. In summary, our study revealed that WAL3 is essential for chloroplast development in rice.

Comparative Transcriptome Analysis Identified Key Pathways and Genes Regulating Differentiated Stigma Color in Melon (Cucumis melo L.)

Stigma color is an important morphological trait in many flowering plants. Visual observations in different field experiments have shown that a green stigma in melons is more attractive to natural pollinators than a yellow one. In the current study, we evaluated the characterization of two contrasted melon lines (MR-1 with a green stigma and M4-7 with a yellow stigma). Endogenous quantification showed that the chlorophyll and carotenoid content in the MR-1 stigmas was higher compared to the M4-7 stigmas. The primary differences in the chloroplast ultrastructure at different developmental stages depicted that the stigmas of both melon lines were mainly enriched with granum, plastoglobulus, and starch grains. Further, comparative transcriptomic analysis was performed to identify the candidate pathways and genes regulating melon stigma color during key developmental stages (S1–S3). The obtained results indicated similar biological processes involved in the three stages, but major differences were observed in light reactions and chloroplast pathways. The weighted gene co-expression network analysis (WGCNA) of differentially expressed genes (DEGs) uncovered a “black” network module (655 out of 5302 genes), mainly corresponding to light reactions, light harvesting, the chlorophyll metabolic process, and the chlorophyll biosynthetic process, and exhibited a significant contribution to stigma color. Overall, the expression of five key genes of the chlorophyll synthesis pathway—CAO (MELO03C010624), CHLH (MELO03C007233), CRD (MELO03C026802), HEMA (MELO03C011113), POR (MELO03C016714)—were checked at different stages of stigma development in both melon lines using quantitative real time polymerase chain reaction (qRT-PCR). The results exhibited that the expression of these genes gradually increased during the stigma development of the MR-1 line but decreased in the M4-7 line at S2. In addition, the expression trends in different stages were the same as RNA-seq, indicating data accuracy. To sum up, our research reveals an in-depth molecular mechanism of stigma coloration and suggests that chlorophyll and related biological activity play an important role in differentiating melon stigma color.

Research on lncRNA related to drought resistance of Shanlan upland rice

BackgroundDrought has become the major abiotic stress that causes losses in rice yields and consequently is one of the main environmental factors threatening food security. Long non-coding RNA (lncRNA) is known to play an important role in plant response to drought stress, while the mechanisms of competing endogenous RNA (ceRNA) in drought resistance in upland rice have been rarely reported.ResultsIn our study, a total of 191 lncRNAs, 2115 mRNAs and 32 miRNAs (microRNAs) were found by strand-specific sequencing and small RNA sequencing to be differentially expressed in drought-stressed rice. Functional analysis of results indicate that they play important roles in hormone signal transduction, chlorophyll synthesis, protein synthesis and other pathways. Construction of a ceRNA network revealed that MSTRG.28732.3 may interact with miR171 in the chlorophyll biosynthesis pathway and affect the ability of plants to withstand drought stress by regulating Os02g0662700, Os02g0663100 and Os06g0105350. The accuracy of the regulatory network was verified by qRT-PCR.ConclusionOur results provide a theoretical basis for future studies on the potential function of lncRNA in plant drought resistance, and they provide new genetic resources for drought-resistant rice breeding.

5-Aminolevulinic acid and hydrogen sulphide alleviate chilling stress in pepper (Capsicum annuum L.) seedlings by enhancing chlorophyll synthesis pathway

5- Aminolevulinic acid (ALA) as a precursor in chlorophyll (Chl) synthesis and hydrogen sulphide (H2S) as a gas signalling molecules can alleviate various abiotic stresses by enhancing photosynthesis. However, little is known about their mechanisms ameliorating photosynthesis under chilling stress, or interactions of ALA and H2S in Chl synthesis. In this study, we explored the effects of exogenous ALA and H2S on chilling stress-induced photosynthesis damage in pepper (Capsicum annuum L.) seedlings. Chilling inhibited the photosynthetic capacity of pepper seedlings, ALA or H2S treatment alone could alleviate this inhibition, and ALA + H2S treatment was even more effective for improving photosynthetic capacity. Additionally, levels of Chl synthesis pathway substances including endogenous ALA, protoporphyrin IX (Proto IX), Mg-protoporphyrin (Mg-Proto IX), protochlorophyllide (Pchl) and Chl (Chl a and Chl b) were significantly decreased, and chilling down-regulated upstream genes HEMA1, HEMB, FAR1, FHY3, CHLH, HEME1, HEMF and PORA. ALA + H2S treatment significantly increased levels of Chl and upstream substances, and up-regulated expression of HEMA1, HEMB and FAR1. In conclusion, exogenous ALA and H2S enhanced chlorophyll synthesis pathway, and thus improved the photosynthesis of pepper seedlings under chilling stress.

Comparative analysis of the metabolome and transcriptome between green and albino zones of variegated leaves from Hydrangea macrophylla ‘Maculata’ infected by hydrangea ringspot virus

In nature, many different factors cause plants to present variegated leaves. The purpose of this study was to reveal the changes in the green and albino leaves of Hydrangea macrophylla ‘Maculata’. It was found that in the albino zone, the leaves became thinner, the chloroplast structure disappeared, and a large number of leucoplasts replaced chloroplasts. In addition, the albino zone of the leaves contained almost no chlorophyll and showed no function related to transforming and utilizing light energy, and more intense oxidative stress was observed in the albino zone of the leaves than in the green zone. RNA-seq analysis showed that the chlorophyll synthesis pathway of the albino zone of leaves was blocked. Upregulated expression of the hydrangea ringspot virus (HdRSV) coat protein (CP) gene was detected in albino tissue by RT-qPCR. Finally, combined UPLC-MS/MS and RNA-seq analyses revealed metabolic changes involving multiple pathways in albino leaf tissue, centered on the TCA cycle. We hypothesize that HdRSV may alter energy metabolism in the albino zone of leaves, including increased lipid metabolism, reduced sugar metabolism, and increased synthesis of amino acids and the viral capsid protein from ribosomes.

Comparative transcriptome and proteome profiling of chlorophyll metabolism pathway in four types of Magnolia officinalis

Magnolia officinalis is a traditional Chinese medicine,with many years of cultivating process, M. officinalis leaves show more differentiation types due to the exchange of seeds from different provenances. "Da Ao"(DA), "Xiao Ao"(XA), "Chuan Hou"(CH),and "Liu Ye"(LY)are the main types of M. officinalis in Sichuan province of China,and there were obvious differences in growth rate,chemical composition,leaf shape and leaf colour. This study selected different types of M. officinalis leaves(DA,XA,LY and CH)from Sichuan to determine their chlorophyll content. Transcriptomic level sequencing of different types of M. officinalis leaf tissues was by high-throughput sequencing analysis and proteomics used an integrated approach involving TMT labelling and LC-MS/MS to quantify the dynamic changes of the whole proteome of M. officinalis. The results showed that CH had the lowest chlorophyll content while DA had the highest chlorophyll content. Furthermore,transcriptome and proteomics results showed that chlorophyll synthesis pathway in DA glutamine-tRNA reductase,urinary porphyrins decarboxylase(UROD),oxygen-dependent protoporphyrin(ODCO),the original-Ⅲ oxidase protoporphyrin oxidase(PPO),magnesium chelating enzyme subunit ChlD,protoporphyrin magnesium Ⅸ monomethyl ester [oxidative] cyclase(MPPMC)were significantly higher than CH,XA and LY,consistent in the results of determination of chlorophyll content(chlorophyll content was highest of 37.56 mg·g~(-1) FW). Some rate-limiting enzymes related to the chlorophyll synthesis,such as ODCO,PPO and MPPMC were tested by Parallel Reaction Monitoring(PRM),and the results showed that the rate-limiting enzyme content in DA was higher than that in other three types. Therefore,based on the differences in leaf color of four types of M. officinalis,the research conducted a preliminary study on the chlorophyll metabolism pathway in leaves of different types of M. officinalis,and explored relevant genes and proteins causing leaf color differences from the molecular level,so as to lay a foundation for studying the differences in growth and development of different types of M. officinalis.

Alleviation of cadmium toxicity by mercapto-triazole priming in wheat

ABSTRACT Cadmium happens to interfere with chlorophyll synthesis, calvin cycle and phosphate pathway and the resulting changes in biochemical attributes induce disorders in physiology, biochemistry and genetics of the germinating plants. Its hazardous effects disturb the phenolic contents, sugars, soluble proteins and activities of antioxidant enzymes. We employed mercapto-triazoles as wheat seed priming agents and evaluated their potential to alleviate the negative impact of cadmium stress (5 µM) on biochemical attributes in germinating seedlings. For priming, 10, 20, 30 and 40 mg L−1 solution of each triazolic compound was used and the germination was carried out under normal and Cd stress conditions. Biochemical attributes including sugars, protein contents, total phenolic, total oxidant status, hydrolytic enzymes and antioxidant enzymes were studied under both conditions and compared with control. Almost all triazolic-priming significantly alleviated the deleterious impacts of cadmium and improved protein contents, SOD, POD, and esterase activities with a significant reduction in MDA contents. The decrease in MDA along with the increase in above-said biochemical attributes in wheat seedlings suggests the mitigating effects of tested triazoles against cadmium stress.

Transcriptome Analysis of Differentially Expressed Genes in Wild Jujube Seedlings under Salt Stress

Wild jujube (Ziziphus acidojujuba) and cultivated jujube (Ziziphus jujuba) belong to the family Rhamnaceae. Jujubes have marked drought- and salt-tolerant properties. After salt stress, wild jujube seedling growth was inhibited and photosynthetic efficiency was reduced. A bioinformatics approach was used to analyze the transcriptomics data from wild jujube seedlings grown under salt stress, and the genes differentially expressed under the salt stress were identified to provide a theoretical basis for the development and use of wild jujube plantations in salinized soil. The transcriptome sequencing from leaves of wild jujube seedlings was carried out using second-generation sequencing technology. The effects of salt stress on the differential expression of photosynthesis-related genes in wild jujube seedlings were analyzed. Transcriptome sequencing revealed a total of 5269 differentially expressed genes (DEGs), of which 2729 were up-regulated and 2540 were down-regulated. DEGs were mainly enriched with respect to photosynthesis, photosynthetic antenna proteins, glyoxylic acid and dicarboxylic acid metabolism, linolenic acid metabolism, cysteine and methionine metabolism, and porphyrin and chlorophyll metabolism. Among them, the photosynthesis pathway-related DEGs were most highly enriched. Further analysis of porphyrin and chlorophyll synthesis and photosynthesis-related pathways revealed that they were significantly enriched by 97 photosynthesis-related DEGs. The DEGs in the photosynthesis and photosynthetic antenna protein pathways were down-regulated, whereas the DEGs glutamyl-tRNA reductase (HEMA), ferrochelatase (HEMH), and pheophorbide a oxygenase (PAO) in the porphyrin and chlorophyll synthesis pathways were up-regulated, with the remainder being down-regulated. The nuclear gene encoding Rubisco, the key enzyme in the photosynthetic carbon fixation pathway, was also down-regulated. The results showed that the photosynthetic rate of wild jujube seedlings decreased following exposure to salinity stress, an effect that was related to the increased synthesis of 5-aminolevulinic acid and heme, and the up-regulation of expression of a gene encoding a chlorophyll-degrading enzyme, and was related to the down-regulation of gene expression in photosynthesis-related pathways such as light energy capture and carbon fixation. Selection of nine DEGs related to photosynthesis and chlorophyll biosynthesis by quantitative real-time-PCR confirmed that expression changes of these nine DEGs were consistent with the transcriptome sequencing results.

Comprehensive study of the genes involved in chlorophyll synthesis and degradation pathways in some monocot and dicot plant species

Chlorophyll (Chl) biosynthesis is one of the most important cellular processes essential for plant photosynthesis. Chl degradation pathway is also important catabolic process occurs during leaf senescence, fruit ripening and under biotic or abiotic stress conditions. Here we have systematically investigated the molecular evolution, gene structure, compositional analysis along with ENc plot, correspondence analysis and codon usage bias of the proteins and encoded genes involved in Chl metabolism from monocots and dicots. The gene and species specific phylogenetic trees using amino acid sequences showed clear clustering formation of the selected species based on monocots and dicots but not supported by 18S rRNA. Nucleotide composition of the encoding genes showed that average GC%, GC1%, GC2% and GC3% were higher in monocots. RSCU analysis depicts that genes from monocots for both pathways and genes for synthesis pathway from dicots only biased to G/C-ending synonymous codons but in degradation pathway most optimal codons (except UUG) in dicots biased to A/U-ending synonymous codons. We found strong evidence of episodic diversifying selection at several amino acid sites in all genes investigated. Conserved domain and gene structures were observed for the genes with varying lengths of introns and exons, involved in Chl metabolism along with some intronless genes within synthesis pathway. ENc and correspondence analyses suggested the mutational or selection constraint on the genes to shape the codon usage. These comprehensive studies may be helpful in further research in molecular phylogenetics and genomics and to better understand the evolutionary dynamics of Chl metabolic pathway. Communicated by Ramaswamy H. Sarma

Genetic dissection of photochemical efficiency under water-deficit stress in rice

Chlorophyll fluorescence (Chl-F) measurements together with non-invasive estimations of chlorophyll content, can be used to investigate functionally rich or poor photosystem II (PSII), relating to alterations in photosynthetic performances under different abiotic stresses. The aim was to identify genetic loci that control rice capacity to cope with different soil moisture conditions such as non-stress (control), water deficit and recovery during the reproductive stage. A genome-wide association study was performed for effective quantum yield of photosystem II (QY) and chlorophyll index across all three treatments. Accessions showed significant variability in traits within each treatment. A total of 43 genetic loci associated with QY and chlorophyll index were identified. Of the total genetic loci identified, 14 were for control, 13 for water-deficit stress and 16 for recovery responses. Interestingly, the majority of the identified genetic loci were co-localized either with chlorophyll synthesis or degradation pathways, components of PSII, transcription factors, protein kinases, transporters, kinases, and antioxidants genes. Favorable alleles and donor accessions found in our study would complement efforts aimed at stacking of traits. Moreover, our results provide promising genetic information for future validation and a potential resource for improving photochemical efficiency and subsequently enhancing carbon gain in rice under water-limited conditions.