Porphyrin Metabolism Research Articles

HFE hemochromatosis gene polymorphism and porphyria cutanea tarda

Purpose. To study the frequency of genotypes and alleles of C282Y and H63D mutations in the HFE gene in patients with porphyria cutanea tarde of the West Siberian region and their influence on the clinical and biochemical characteristics of the disease. Materials and methods. We observed 14 patients with porphyria cutanea tarde (PCT), who underwent a comprehensive general clinical and instrumental examination. The indicators of the excretory profile of porphyrin metabolism indicators were purposefully determined, a molecular genetic examination was carried out to determine the genotypes and alleles of the C282Y and H63D mutations of the hemochromatosis gene HFE. Results and discussion. Molecular genetic research found that the HFE gene mutation was found in 7 patients (50.0%). Polymorphism for the C282Y allele was found in 2 (14.3%) patients, and for the H63D allele - in 5 patients (35.7%). Risk factors for the manifestation of PCT included frequent alcohol consumption and chronic HCV infection. Genotype 1b was recorded more often (9 people), less often - genotype 3a (4 people). The excretory profile of porphyrin metabolism indices in patients of both groups exceeded the control values. Porphyrin metabolism parameters in patients without HFE gene mutations were significantly higher than those in patients without HFE gene mutations. Conclusions. Polymorphism of the hemochromatosis gene HFE in PCT was detected in 50% of patients. The most common mutation was the H63D allele. The level of porphyrin metabolism disorders in patients with HFE hemochromatosis gene mutations is significantly lower. The clinical picture of PCT in all observed patients did not differ. Chronic viral hepatitis C is assessed as a risk factor for the manifestation of PCT.

Transcriptional profiling of pigmentation-regulated long non-coding RNAs in the mantle of Manila clam (Ruditapes philippinarum)

Long non-coding RNAs (lncRNAs) play important roles in a variety of biological processes and have received extensive attention in recent years. However, the expression and roles of lncRNAs in the Manila clam (Ruditapes philippinarum) are still poorly understood. In this study, we identified lncRNAs in the mantle of clams with different shell colors (orange (O), zebra (Z), white (W), and white zebra (WZ)). Using poly(A)-independent and strand-specific RNA-seq, we obtained 183,818,4888 clean reads and 32,541 lncRNA transcripts. In total, 1988 lncRNAs were identified as differentially expressed lncRNAs in comparisons between six pairs of groups. Fourteen mRNAs and their target genes were found to be related to the shell color pathway of melanin and porphyrin metabolism. Protein kinase C, calmodulin, melanogenesis associated transcription factor, peroxidase-like protein, tyrosine decarboxylase, myeloperoxidase, putativetyrosinase-like protein tyr-3, dopamine beta-hydroxylase, 5-aminolevulinate synthase, tyrosine aminotransferase, uroporphyrinogen-III synthase, hephaestin, delta-aminolevulinic acid dehydratase and ferrochelatase and their corresponding 77 cis-acting lncRNAs are thought to play important roles in the melanin synthesis pathway and porphyrin metabolic pathway. This study identified the catalog of lncRNAs expressed in the mantle of Manila clam, and the results provide insights into understanding the molecular regulation of shell color and genetic breeding in R. philippinarum.

Overexpression of 2-Cys Peroxiredoxin alleviates the NaHCO3 stress-induced photoinhibition and reactive oxygen species damage of tobacco

Plant 2-cysteine peroxiredoxin (2-Cys Prx) is a mercaptan peroxidase localized in chloroplasts and has unique catalytic properties. To explore the salt stress tolerance mechanisms of 2-Cys Prx in plants, we analyzed the effects of overexpressing the 2-CysPrx gene on the physiological and biochemical metabolic processes of tobacco under NaHCO3 stress through joint physiological and transcriptomic analysis. These parameters included growth phenotype, chlorophyll, photosynthesis, and antioxidant system. After NaHCO3 stress treatment, a total of 5360 differentially expressed genes (DEGs) were identified in 2-Cysprx overexpressed (OE) plants, and the number of DEGs was significantly lower than 14558 in wild-type (WT) plants. KEGG enrichment analysis showed that DEGs were mainly enriched in photosynthetic pathways, photosynthetic antenna proteins, and porphyrin and chlorophyll metabolism. Overexpressing 2-CysPrx significantly reduced the growth inhibition of tobacco induced by NaHCO3 stress, alleviating the down-regulation of the DEGs related to chlorophyll synthesis, photosynthetic electron transport and the Calvin cycle and the up-regulation of those related to chlorophyll degradation. In addition, it also interacted with other redox systems such as thioredoxins (Trxs) and the NADPH-dependent Trx reductase C (NTRC), and mediated the positive regulation of the activities of antioxidant enzymes such as peroxidase (POD) and catalase (CAT) and the expression of related genes, thereby reducing the accumulation of superoxide anion (O2·-), hydrogen peroxide (H2O2) and malondialdehyde (MDA). In conclusion, 2-CysPrx overexpression could alleviate the NaHCO3 stress-induced photoinhibition and oxidative damage by regulating chlorophyll metabolism, promoting photosynthesis and participating in the regulation of antioxidant enzymes, and thus improve the ability of plants to resist salt stress damage.

Analisis Perbandingan Certainty Factor dan Dempster Shafer Dalam Diagnosis Penyakit Porfiria Menggunakan Metode Perbandingan Eksponensial

Porphyria refers to a group of genetic disorders that affect the metabolism of porphyrins in the body. Porphyrin itself is a molecule that plays a role in the synthesis of hemoglobin, which functions as a carrier of oxygen in the blood. Disturbances in porphyrin metabolism can lead to the accumulation of porphyrins or their precursors, which in turn can produce a variety of symptoms. Porphyria comes in many forms, including acute porphyria and skin porphyria, each with its own unique symptoms. Nervous system and skin problems are common in porphyria patients, and if not treated properly, their health condition can deteriorate. A unique approach is needed to provide answers to this problem because it is still challenging to develop practical ways to address the problems experienced by patients. Certainty Factor and Dempster Shafer methods are two techniques that are widely used in the field of disease diagnosis today, where expert systems are often applied. But building expert systems in multiple disciplines is difficult due to uncertainty. Therefore, it is important to research and distinguish the many ways that these systems can be built. The exponential comparison approach is one of the most straightforward comparison techniques and helps minimize bias in the analysis process. To identify candidiasis in humans, this study attempted to apply, evaluate, and compare the two methodologies, as well as compare the results with the exponential comparison method. Comparative findings suggest that the Dempster Shafer technique provides a more precise diagnosis of porphyria

Exploring the effect of the Uyghur medicine Munziq Balgam on a collagen-induced arthritis rat model by UPLC-MS/MS-based metabolomics approach.

Munziq Balgam (MBm) is a classic preparation of a traditional Uyghur medicine used for many years to treat abnormal body fluid diseases. The formula, as an in-hospital preparation, has already been used in the Hospital of Xinjiang Traditional Uyghur Medicine to treat rheumatoid arthritis (RA) with significant clinical effects. This study intends to reveal the intervention effect of MBm on collagen-induced arthritis (CIA) rats, discover the potential biomarkers with efficacy, and explore the mechanisms of metabolic regulation by using metabolomics method. Sprague Dawley (SD) rats were randomly divided into five groups: blank group, CIA model group, Munziq Balgam nomal-dosage, Munziq Balgam high-dosage group and control group. Body weight, paw swelling, arthritis index, immune indices and histopathological experiments were carried out. Plasma from rats were detected by UPLC-MS/MS. Metabolomics of plasma was performed to analyze metabolic profiles, potential biomarkers, and metabolic pathways of MBm for CIA rats. The main metabolic result of Uyghur medicine MBm was compared with that of Zhuang medicine Longzuantongbi granules (LZTBG) to explore the characteristics of two ethnic medicines from different regions for RA. MBm could significantly alleviate symptoms of CIA rats by relieving arthritis symptoms on paw redness and swelling, inflammatory cell infiltration, synovial hyperplasia, pannus, cartilage and bone tissue destruction, as well as inhibiting the expression of IL-1β, IL-6, TNF-α, UA and ALP. Linoleic acid, alpha-linolenic acid, pantothenate and CoA biosynthesis, achidonic acid, gycerophospholipid, sphingolipid metabolism, primary bile acid biosynthesis, porphyrin and chlorophyll metabolism and fatty acid degradation served as the main nine pathways of the interventional effect of MBm on CIA rats. Twenty-three different metabolites were screened out and strongly associated with the indicator makes of RA. Eight potential efficacy-related biomarkers were finally discovered in metabolic pathway network (phosphatidylcholine, bilirubin, sphinganine 1-phosphate, phytosphingosine, SM (d18:1/16:0), pantothenic acid, l-palmitoylcarnitine, chenodeoxycholate). Three metabolites (chenodeoxycholate, hyodeoxycholic acid and O-palmitoleoylcarnitine) were changed in both the metabolic study of MBm and LZTBG intervention effects on CIA rats. Additionally, MBm and LZTBG shared the same 6 metabolic pathways including linoleic acid, alpha-linolenic acid, pantothenate and CoA biosynthesis, achidonic acid, gycerophospholipid, and primary bile acid biosynthesis. The study suggested that MBm may effectively alleviate RA by regulating inflammation, immunity-related pathways and multiple targets. Metabolomics analysis showed that MBm (Xinjiang, the north of China) and LZTBG (Guangxi, the south of China), two ethnic medicines from different regions in China, share common metabolites and pathways but also have distinct differences in their interventions for RA.

Leaf Color Classification and Expression Analysis of Photosynthesis-Related Genes in Inbred Lines of Chinese Cabbage Displaying Minor Variations in Dark-Green Leaves.

The leaves of the Chinese cabbage which is most widely consumed come in a wide variety of colors. Leaves that are dark green can promote photosynthesis, effectively improving crop yield, and therefore hold important application and cultivation value. In this study, we selected nine inbred lines of Chinese cabbage displaying slight differences in leaf color, and graded the leaf color using the reflectance spectra. We clarified the differences in gene sequences and the protein structure of ferrochelatase 2 (BrFC2) among the nine inbred lines, and used qRT-PCR to analyze the expression differences of photosynthesis-related genes in inbred lines with minor variations in dark-green leaves. We found expression differences among the inbred lines of Chinese cabbage in photosynthesis-related genes involved in the porphyrin and chlorophyll metabolism, as well as in photosynthesis and photosynthesis-antenna protein pathway. Chlorophyll b content was significantly positively correlated with the expression of PsbQ, LHCA1_1 and LHCB6_1, while chlorophyll a content was significantly negatively correlated with the expression PsbQ, LHCA1_1 and LHCA1_2. Our results provide an empirical basis for the precise identification of candidate genes and a better understanding of the molecular mechanisms responsible for the production of dark-green leaves in Chinese cabbage.

Heterologous expression of Zygophyllum xanthoxylon zinc finger protein gene (ZxZF) enhances the tolerance of poplar photosynthetic function to drought stress

The ZxZF transcription factor (TF) of Zygophyllum xanthoxylon (Bunge) Maxim, an extremely drought-resistant woody plant, is a C2H2 zinc finger protein. Studies have shown that C2H2 zinc finger proteins play important roles in activating stress-related genes and enhancing plant resistance. However, their function in regulating plant photosynthesis under drought stress is not well understood. Since poplar is an important greening and afforestation tree species, it is particularly important to cultivate excellent drought-tolerant varieties. The ZxZF transcription factor (TF) was heterogeneously expressed in Euroamerican poplar (Populus × euroameracana cl.'Bofengl’) by genetic transformation. Based on the mechanism and potential function of poplar photosynthesis regulated by ZxZF under drought stress, transcriptomic and physiological determinations were used to reveal the important role of this gene in improving the drought resistance of poplar. The results showed that the overexpression of ZxZF TF in transgenic poplars could improve the inhibition of Calvin cycle by regulating stomatal opening and increasing the concentration of intercellular CO2. The chlorophyll content, photosynthetic performance index, and photochemical efficiency of transgenic lines under drought stress were significantly higher than those of the wild type (WT). The overexpression of ZxZF TFs could alleviate the degree of photoinhibition of photosystems II and I under drought stress and maintain the efficiency of light energy capture and the photosynthetic electron transport chain. The transcriptomic data also showed that differentially expressed genes between the transgenic poplar and WT under drought stress were primarily enriched in metabolic pathways related to photosynthesis, such as photosynthesis, photosynthesis-antenna protein, porphyrin and chlorophyll metabolism, and photosynthetic carbon fixation, and the downregulation of genes related to chlorophyll synthesis, photosynthetic electron transport and Calvin cycle were alleviated. In addition, the overexpression of ZxZF TF can alleviate the inhibition of NADH dehydrogenase-like (NDH) cyclic electron flow of the poplar NDH pathway under drought stress, which plays an important role in reducing the excess pressure of electrons on the photosynthetic electron transport chain and maintaining the normal photosynthetic electron transport. In summary, the overexpression of ZxZF TFs can effectively alleviate the inhibition of drought on the assimilation of carbon in poplar and have a positive impact on light energy capture, the orderly transport of photosynthetic electron transport chain and the integrity of the photosystem, which is highly significant to acheivean in-depth understanding of the function of ZxZF TFs. This also provides an important basis for the breeding of new transgenic poplar varieties.

CsXDH1 gene promotes caffeine catabolism induced by continuous strong light in tea plant.

Tea plant (Camellia sinensis) is an important cash crop with extensive adaptability in the world. However, complex environmental factors force a large variation of tea quality-related components. Caffeine is essential for the formation of bitter and fresh flavors in tea, and is the main compound of tea that improves human alertness. Continuous strong light stimulation was observed to cause caffeine reduction in tea leaves, but the mechanism is not clear. In this study, the response of tea plant to light intensity was analysed mainly by multi-omics association, antisense oligodeoxynucleotide (asODN) silencing technique, and in vitro enzyme activity assay. The results revealed multiple strategies for light intensity adaptation in tea plant, among which the regulation of chloroplasts, photosynthesis, porphyrin metabolism, and resistance to oxidative stress were prominent. Caffeine catabolism was enhanced in continuous strong light, which may be a light-adapted strategy due to strict regulation by xanthine dehydrogenase (XDH). asODN silencing and enzymatic activity assays confirmed that CsXDH1 is a protein induced by light intensity to catalyze the substrate xanthine. CsXDH1 asODN silencing resulted in significant up-regulation of both caffeine and theobromine in in vitro enzyme activity assay, but not in vivo. CsXDH1 may act as a coordinator in light intensity adaptation, thus disrupting this balance of caffeine catabolism.

Response of Salmonella enterica serovar Typhimurium to alginate oligosaccharides fermented with fecal inoculum: integrated transcriptomic and metabolomic analyses

Alginate oligosaccharides (AOS), extracted from marine brown algae, are a common functional feed additive; however, it remains unclear whether they modulate the gut microbiota and microbial metabolites. The response of Salmonella enterica serovar Typhimurium, a common poultry pathogen, to AOS fermented with chicken fecal inocula was investigated using metabolomic and transcriptomic analyses. Single-strain cultivation tests showed that AOS did not directly inhibit the growth of S. Typhimurium. However, when AOS were fermented by chicken fecal microbiota, the supernatant of fermented AOS (F-AOS) exhibited remarkable antibacterial activity against S. Typhimurium, decreasing the abundance ratio of S. Typhimurium in the fecal microbiota from 18.94 to 2.94%. Transcriptomic analyses showed that the 855 differentially expressed genes induced by F-AOS were mainly enriched in porphyrin and chlorophyll metabolism, oxidative phosphorylation, and Salmonella infection-related pathways. RT-qPCR confirmed that F-AOS downregulated key genes involved in flagellar assembly and the type III secretory system of S. Typhimurium, indicating metabolites in F-AOS can influence the growth and metabolism of S. Typhimurium. Metabolomic analyses showed that 205 microbial metabolites were significantly altered in F-AOS. Among them, the increase in indolelactic acid and 3-indolepropionic acid levels were further confirmed using HPLC. This study provides a new perspective for the application of AOS as a feed additive against pathogenic intestinal bacteria.

Effects of Magnesium on Transcriptome and Physicochemical Index of Tea Leaves.

Magnesium (Mg) is one of the essential elements for the growth of tea tree and is extremely important for its development. In this study, we investigated the effect of Mg on the transcriptome and physicochemical indexes of tea leaves, and the results showed that Mg could significantly affect the gene expression of tea leaves. The results of Orthogonal Partial Least-Squares Discriminant Analysis (OPLS-DA) model analysis showed that a total of 300 key genes (Variable Importance for the Projection, VIP > 1) were screened under different concentrations of Mg treatment, among which 140 genes were up-regulated and 160 genes were down-regulated. The bubble map was used to screen the characteristic genes from the above key genes, and a total of 121 representative characteristic genes were obtained, mainly involving 9 metabolic pathways. Among them, gene expression of three metabolic pathways, including porphyrin metabolism, alpha-linolenic acid metabolism and photosynthesis, showed an increasing trend with the increase of Mg concentration, while gene expression of four metabolic pathways, including biosynthesis of secondary metabolites, anthocyanin biosynthesis, ABC transporters, pentose and glucuronate interconversions, showed a decreasing trend. The results of physiological index analysis showed that with the increase of Mg concentration, the photosynthetic physiological index, theanine and soluble sugar content of tea leaves showed an increasing trend, while the content of tea polyphenol, flavone and caffeine showed a decreasing trend. The results of TOPSIS analysis showed that the physiological indexes of tea trees most affected by Mg were chlorophyll, tea polyphenols and flavonoids, while the metabolic pathways most affected by Mg on gene expression were the metabolic pathways and biosynthesis of secondary metabolites. It can be seen that the effects of Mg on tea tree were mainly related to photosynthesis and synthesis of secondary metabolites, and Mg was beneficial for improving the photosynthetic capacity of tea tree, enhancing the accumulation of primary metabolites, and thus increasing tea yield. However, Mg was not conducive to the synthesis of secondary metabolites of tea tree and the accumulation of main quality indexes of tea leaves.

Cell senescence-associated porphyrin metabolism affects the efficacy of aminolevulinic acid-photodynamic diagnosis in bladder cancer

Aminolevulinic acid-photodynamic diagnosis (ALA-PDD) is a promising alternative method to detect cancer cells because of its high specificity and low rate of side effects. Exogenous ALA is administered and accumulates as protoporphyrin IX (PpIX) in cancer cells, which then emit red fluorescence following light irradiation to enable surgeons to accurately identify and remove cancerous tissue. Recent reports suggested that PpIX failed to accumulate in some patients who underwent ALA-PDD. We hypothesized that cell senescence, which is a relatively inactive state, affects porphyrin accumulation in bladder cancer cells. In this study, we evaluated the relationship between cell senescence and porphyrin accumulation in affecting the efficacy of ALA-PDD. First, we utilized three bladder cancer cell lines to evaluate senescence-related indicators and establish a cell senescence model. Then, we identified the differences in porphyrin production and the proteins involved in porphyrin accumulation between old and young cells. We found that compared with young cells, old cells possessed higher concentration of PpIX and had lower ABCG2 expression. The increase in PpIX levels following ABCG2 inhibition is three times higher in old cells than in young cells, suggesting that cell senescence was closely related with porphyrin accumulation in cancer. In conclusion, we found that the efficacy of ALA-PDD and porphyrin accumulation was relatively high in senescent cancer cells and that inhibition of ABCG2 could improve the efficacy of ALA-PDD in young bladder cancer cells.

Responses of differential metabolites and pathways to high temperature in cucumber anther.

Cucumber is one of the most important vegetable crops, which is widely planted all over the world. Cucumber always suffers from high-temperature stress in South China in summer. In this study, liquid chromatography-mass spectrometry (LC-MS) analysis was used to study the differential metabolites of cucumber anther between high-temperature (HT) stress and normal condition (CK). After HT, the pollen fertility was significantly reduced, and abnormal anther structures were observed by the paraffin section. In addition, the metabolomics analysis results showed that a total of 125 differential metabolites were identified after HT, consisting of 99 significantly upregulated and 26 significantly downregulated metabolites. Among these differential metabolites, a total of 26 related metabolic pathways were found, and four pathways showed significant differences, namely, porphyrin and chlorophyll metabolism; plant hormone signal transduction; amino sugar and nucleotide sugar metabolism; and glycine, serine, and threonine metabolism. In addition, pollen fertility was decreased by altering the metabolites of plant hormone signal transduction and amino acid and sugar metabolism pathway under HT. These results provide a comprehensive understanding of the metabolic changes in cucumber anther under HT.

PORPHYRIA CUTANEATARDA: A CASE REPORT

Porphyria cutaneatarda (PCT) is a rare disease of porphyrin metabolism, related to a deficiency of Uroporphyrinogen decarboxylase activity. Clinically, it manifests itself by a skin fragility in photo-exposed areas. The characteristic biochemical profile of PCT, with elevated levels of urinary and plasma porphyrins, establishes the diagnosis. Treatment is based on phlebotomy, hydroxychloroquine (100 to 200 mg twice weekly) and control of susceptibility factors. We report a case of porphyria cutaneatarda in a male coast guard by profession.

Effects of histone methylation modification on low temperature seed germination and growth of maize

Low temperature is a limiting factor of seed germination and plant growth. Although there is a lot information on the response of maize to low temperatures, there is still poorly description of how histone methylation affects maize germination and growth development at low temperatures. In this study, the germination rate and physiological indexes of wild-type maize inbred lines B73 (WT), SDG102 silencing lines (AS), SDG102 overexpressed lines (OE) at germination stage and seedling stage were measured under low temperature stress (4 ℃), and transcriptome sequencing was applied to analyze the differences of gene expression in panicle leaves among different materials. The results showed that the germination rate of WT and OE maize seeds at 4 ℃ was significantly lower than 25 ℃. The content of MDA, SOD and POD of 4 ℃ seeding leaves higher than contrast. Transcriptome sequencing results showed that there were 409 different expression genes (DEGs) between WT and AS, and the DEGs were mainly up-regulated expression in starch and sucrose metabolism and phenylpropanoid biosynthesis. There were 887 DEGs between WT and OE, which were mainly up-regulated in the pathways of plant hormone signal transduction, porphyrin and chlorophyll metabolism. This result could provide a theoretical basis for analyzing the growth and development of maize from the perspective of histone methylation modification.

Photosensitization dermatitis in animals

Photosensitization, also known as photodermatitis, is an abnormal reaction of the skin when exposed to light and is caused by accumulation of photosensitizing substances in the skin, resulting in a severe dermatitis of the skin unprotected by unpigmented. There are three types of photosensitization: (a) primary, in which photodynamic chemicals, or plant toxins, reach the skin through the circulation, (b) congenital porphyria, in which there is a metabolic, congenital defect in porphyrin metabolism, and (c) secondary (hepatogenous), which occurs when the capacity of the liver to excrete derived from the catabolism of alimentary chlorophyll is impaired. We consider that liver pathologies are the most important in animals from Romania.

Transcriptome profiling reveals characteristics of hairy root and the role of AhGLK1 in response to drought stress and post-drought recovery in peanut

BackgroundHR (hairy root) has emerged as a valuable tissue for the rapid characterization of plant gene function and enzyme activity in vivo. AhGLK1 (Arachis hypogaea L. golden2-like 1) is known to play a role in post-drought recovery. However, it is unclear (a) whether HR has properties that are distinct from those of PR (primary root); and (b) which gene networks are regulated by AhGLK1 in response to drought stress and recovery in peanut.ResultsWe found that cells of the root tip cortex were larger in HR than in PR, while a total of 850 differentially expressed genes (DEGs) were identified in HR compared to PR. Eighty-eight of these DEGs, relating to chlorophyll and photosynthesis, were upregulated in HR. In addition, AhGLK1-OX (AhGLK1-overexpressing) HR showed a green phenotype, and had a higher relative water content than 35 S::eGFP (control) HR during drought stress. RNA-seq analysis showed that 74 DEGs involved both in the drought response and the post-drought recovery process were significantly enriched in the galactose metabolism pathway. GO terms enrichment analysis revealed that 59.19%, 29.79% and 17.02% of the DEGs mapped to the ‘biological process’ (BP), ‘molecular function’ (MF) and ‘cellular component’ (CC) domains, respectively. Furthermore, 20 DEGs involved in post-drought recovery were uniquely expressed in AhGLK1-OX HR and were significantly enriched in the porphyrin metabolism pathway. GO analysis showed that 42.42%, 30.30% and 27.28% of DEGs could be assigned to the BP, MF and CC domains, respectively. Transcription factors including bHLH and MYB family members may play a key role during drought stress and recovery.ConclusionOur data reveal that HR has some of the characteristics of leaves, indicating that HR is suitable for studying genes that are mainly expressed in leaves. The RNA-seq results are consistent with previous studies that show chlorophyll synthesis and photosynthesis to be critical for the role of AhGLK1 in improving post-drought recovery growth in peanut. These findings provide in-depth insights that will be of great utility for the exploration of candidate gene functions in relation to drought tolerance and/or post-drought recovery ability in peanut.

Proteomic analysis of the initial wake up of vibrio splendidus persister cells

Vibrio splendidus is a ubiquitous pathogen that causes various diseases in aquaculture with a wide range of hosts. In our previous studies, we showed that L-glutamic acid was the optimal carbon source that could revive V. splendidus persister cells. In our present study, single cell observation under microscopy showed that V. splendidus could revive using L-glutamic acid as carbon source. A proteomic analysis was carried out to further illustrate the initial wake up of persister cells with L-glutamic acid. To collect the initially revived cells, SDS-PAGE was used to determine the revived time. The total proteins from the persister cells and the revived cells were analyzed using LC‒MS/MS. A total of 106 proteins, including 42 downregulated proteins and 64 upregulated proteins, were identified. GO analysis of the differentially expressed proteins (DEPs) showed that biological processes, including protein complex assembly, protein oligomerization, and arginine metabolism; cellular components, including extracellular membrane, plasma membrane and ribosome; and molecular functions, including the activities of arginine binding and structural constituent of ribosome, were enriched. KEGG analysis showed that lipopolysaccharide biosynthesis, porphyrin and chlorophyll metabolism, and peptidoglycan biosynthesis were upregulated, while the ribosome was downregulated. This is the first time to study the initial wake up of persister cells based on proteomic analysis, and the results revealed the main pathways involved in the early resuscitation of V. splendidus persister cells.

Identification of circular RNAs in tea plant during Helopeltis theivora infestation

The tea mosquito bug (TMB) or Helopeltis theivora has been reported as a significant tea pest that causes major tea crop loss. Circular RNAs (circRNAs) are a newly discovered group of the non-coding RNA (ncRNA) and are known to function in gene regulation process. Tea plant (Camellia sinensis) has not been explored for the detection of circRNAs during pest infestation. This is the first report of circRNAs identified in C. sinensis infested by a sap-sucking insect H. theivora. In total, we identified 709 circRNAs in six RNA-seq libraries. Analysis of the differential expression pattern revealed that 34 circRNAs got upregulated/downregulated in response to H. theivora attack. Functional annotation and enrichment analysis of the differentially expressed circRNAs showed that the circRNA-target genes were enriched in biological pathways like “tryptophan metabolism”, “biosynthesis of secondary metabolites”, “porphyrin metabolism”, “nucleocytoplasmic transport”, “N-glycan biosynthesis” etc. miRNA-mediated interaction analysis of the circRNAs also showed that 17 differentially expressed circRNAs could also act as “sponges” thus masking the miRNA-mRNA binding and subsequently repressing mRNA cleavage. These differentially expressed circRNAs were predicted to have the capacity to behave as endogenous target mimics (eTMs) for potential genes like aspartyl protease, phospholipase, lectin receptor etc.

Function of ALA Content in Porphyrin Metabolism Regulation of Ananas comosus var. bracteatus.

Chlorophyll and heme are essential molecules for photosynthesis and respiration, which are competing branches of the porphyrin metabolism pathway. Chlorophyll and heme balance regulation is very important for the growth and development of plants. The chimeric leaves of Ananas comosus var. bracteatus were composed of central photosynthetic tissue (PT) and marginal albino tissue (AT), which were ideal materials for the study of porphyrin metabolism mechanisms. In this study, the regulatory function of ALA content on porphyrin metabolism (chlorophyll and heme balance) was revealed by comparing PT and AT, 5-Aminolevulinic Acid (ALA) exogenous supply, and interference of hemA expression. The AT remained similar in porphyrin metabolism flow level to the PT by keeping an equal ALA content in both tissues, which was very important for the normal growth of the chimeric leaves. As the chlorophyll biosynthesis in AT was significantly inhibited, the porphyrin metabolism flow was directed more toward the heme branch. Both tissues had similar Mg2+ contents; however, Fe2+ content was significantly increased in the AT. The chlorophyll biosynthesis inhibition in the white tissue was not due to a lack of Mg2+ and ALA. A 1.5-fold increase in ALA content inhibited chlorophyll biosynthesis while promoting heme biosynthesis and hemA expression. The doubling of ALA content boosted chlorophyll biosynthesis while decreasing hemA expression and heme content. HemA expression interference resulted in a higher ALA content and a lower chlorophyll content, while the heme content remained at a relatively low and stable level. Conclusively, a certain amount of ALA was important for the stability of porphyrin metabolism and the normal growth of plants. The ALA content appears to be able to regulate chlorophyll and heme content by bidirectionally regulating porphyrin metabolism branch direction.

Expression of the Arabidopsis Mg-chelatase H subunit alleviates iron deficiency-induced stress in transgenic rice.

The most common symptom of iron (Fe) deficiency in plants is leaf chlorosis caused by impairment of chlorophyll biosynthesis. Magnesium (Mg)-chelatase H subunit (CHLH) is a key component in both chlorophyll biosynthesis and plastid signaling, but its role in Fe deficiency is poorly understood. Heterologous expression of the Arabidopsis thaliana Mg-chelatase H subunit gene (AtCHLH) increased Mg-chelatase activity by up to 6-fold and abundance of its product, Mg-protoporphyrin IX (Mg-Proto IX), by 60-75% in transgenic rice (Oryza sativa) seedlings compared to wild-type (WT) controls. Noticeably, the transgenic seedlings showed alleviation of Fe deficiency symptoms, as evidenced by their less pronounced leaf chlorosis and lower declines in shoot growth, chlorophyll contents, and photosynthetic efficiency, as indicated by F v/F m and electron transport rate, compared to those in WT seedlings under Fe deficiency. Porphyrin metabolism was differentially regulated by Fe deficiency between WT and transgenic seedlings, particularly with a higher level of Mg-Proto IX in transgenic lines, showing that overexpression of AtCHLH reprograms porphyrin metabolism in transgenic rice. Leaves of Fe-deficient transgenic seedlings exhibited greater upregulation of deoxymugineic acid biosynthesis-related genes (i.e., NAS, NAS2, and NAAT1), YSL2 transporter gene, and Fe-related transcription factor genes IRO2 and IDEF2 than those of WT, which may also partly contribute to alleviating Fe deficiency. Although AtCHLH was postulated to act as a receptor for abscisic acid (ABA), exogenous ABA did not alter the phenotypes of Fe-deficient WT or transgenic seedlings. Our study demonstrates that modulation of porphyrin biosynthesis through expression of AtCHLH in transgenic rice alleviates Fe deficiency-induced stress, suggesting a possible role for CHLH in Fe deficiency responses.