Phospholipase D Gene Research Articles

Genome-wide identification of the Phospholipase D (PLD) gene family in Chinese white pear (Pyrus bretschneideri) and the role of PbrPLD2 in drought resistance

The Chinese white pear (Pyrus bretschneideri), a vital fruit crop, is highly susceptible to abiotic stresses, especially drought, which poses a major threat to its growth and productivity. Phospholipase D (PLD) genes are pivotal in orchestrating plant responses to abiotic stresses, acting as key regulators in stress adaptation mechanisms. This study systematically identified and functionally characterized the entire PLD gene family in P. bretschneideri through a comprehensive genome-wide analysis. A total of 20 PbrPLD genes were identified, and they were categorized into five subfamilies based on phylogenetic analysis. chromosome localization, gene structure, and conserved motif analyses revealed that these genes have diverse evolutionary histories. Cis-acting element analysis and expression profiling under drought stress indicated that several PbrPLD genes, particularly PbrPLD2, are strongly induced by drought. Overexpression of PbrPLD2 in both Arabidopsis thaliana and pear demonstrated enhanced drought tolerance through improved stomatal closure and increased expression of drought-responsive genes. These findings highlight the critical role of PbrPLD2 in drought resistance and provide a theoretical and experimental foundation for molecular breeding in pear and other fruit crops.

Knockout of a PLD gene in Schizochytrium limacinum SR21 enhances docosahexaenoic acid accumulation by modulation of the phospholipid profile

BackgroundThe hydrolysis and transphosphatidylation of phospholipase D (PLD) play important roles in the interconversion of phospholipids (PLs), which has been shown to profoundly impact lipid metabolism in plants. In this study, the effect of the PLD1 gene of Schizochytrium limacinum SR21 (S. limacinum SR21) on lipid metabolism was investigated.ResultsPLD1 knockout had little impact on cell growth and lipid production, but it significantly improved the percentage of polyunsaturated fatty acids in lipids, of which docosahexaenoic acid (DHA) content increased by 13.3% compared to the wild-type strain. Phospholipomics and real-time quantitative PCR analysis revealed the knockout of PLD1 reduced the interexchange and increased de novo synthesis of PLs, which altered the composition of PLs, accompanied by a final decrease in phosphatidylcholine (PC) and an increase in phosphatidylinositol, lysophosphatidylcholine, and phosphatidic acid levels. PLD1 knockout also increased DHA content in triglycerides (TAGs) and decreased it in PLs.ConclusionsThese results indicate that PLD1 mainly performs the transphosphatidylation activity in S. limacinum SR21, and its knockout promotes the migration of DHA from PLs to TAGs, which is conducive to DHA accumulation and storage in TAGs via an acyl CoA-independent pathway. This study provides a novel approach for identifying the mechanism of DHA accumulation and metabolic regulation strategies for DHA production in S. limacinum SR21.

Storability, longevity and rancidity: A genomic perspective on evolutionary constraints in major oilseeds

Limited longevity and accelerated chemical decomposition during storage are major problems found in oilseeds. In the present study the genomic composition of rancidity prone soybean and sunflower are compared with oil seeds with fairly good storability like sesame and olive. The chromosomal location, gene duplications, syntenic relationships, cis-element architecture and protein-protein interactions were studied with respect to 250 genes affecting lipid decomposition like lipases, lipoxygenases and oleosins. The expansion of lipoxygenases and phospholipaseD genes in soybean and sunflower indicate the importance of gene duplication in functional divergence. Moreover the effect of strong purifying selection in lipid decomposition genes was evident in shaping the genomic diversity.

Genome-wide identification, structural homology analysis, and evolutionary diversification of the phospholipase D gene family in the venom gland of three scorpion species

BackgroundVenom phospholipase D (PLDs), dermonecrotic toxins like, are the major molecules in the crude venom of scorpions, which are mainly responsible for lethality and dermonecrotic lesions during scorpion envenoming. The purpose of this study was fivefold: First, to identify transcripts coding for venom PLDs by transcriptomic analysis of the venom glands from Androctonus crassicauda, Hottentotta saulcyi, and Hemiscorpius lepturus; second, to classify them by sequence similarity to known PLDs and motif extraction method; third, to characterize scorpion PLDs; fourth to structural homology analysis with known dermonecrotic toxins; and fifth to investigate phylogenetic relationships of the PLD proteins.ResultsWe found that the venom gland of scorpions encodes two PLD isoforms: PLD1 ScoTox-beta and PLD2 ScoTox-alpha I. Two highly conserved regions shared by all PLD1s beta are GAN and HPCDC (HX2PCDC), and the most important conserved regions shared by all PLD2s alpha are two copies of the HKDG (HxKx4Dx6G) motif. We found that PLD1 beta is a 31–43 kDa acidic protein containing signal sequences, and PLD2 alpha is a 128 kDa basic protein without known signal sequences. The gene structures of PLD1 beta and PLD2 alpha contain 6 and 21 exons, respectively. Significant structural homology and similarities were found between the modeled PLD1 ScoTox-beta and the crystal structure of dermonecrotic toxins from Loxosceles intermedia.ConclusionsThis is the first report on identifying PLDs from A. crassicauda and H. saulcyi venom glands. Our work provides valuable insights into the diversity of scorpion PLD genes and could be helpful in future studies on recombinant antivenoms production.

Comparative Analysis of Phospholipase D (PLD) Gene Family in Camelina sativa and Brassica napus and Its Responses in Camelina Seedlings under Salt Stress

Phospholipases are among the important elements involved in lipid-dependent cell signaling that lead to the induction of downstream pathways. In the current study, phospholipases D (PLDs) gene family was characterized and compared in two important oilseed crops, Brassica napus and Camelina sativa. The results revealed that PLD has 33 members in Camelina sativa (CsPLD) and 41 members in Brassica napus (BnPLD). All studied PLDs showed a negative GRAVY value, indicating that PLDs are probably hydrophilic proteins. Phylogenetic analysis classified PLDs into five main subfamilies, including gamma, delta, beta, alpha, and zeta. According to evolution analysis, a different evolution process was observed between CsPLD and BnPLD. In addition, the results disclosed that most of the PLD genes have been segmentally duplicated under purifying selection. Cis-regulatory elements related to ABA and auxin responsiveness were found more in the upstream region of CsPLDs, while elements linked with MeJA responsiveness were observed more in the promoter region of BnPLDs. Analysis of the expression data showed that PLD alpha genes have a wide expression in most tissues. Quantitative expression analysis (qPCR) of CsPLD genes under salt stress, 200 mM of NaCl, was conducted in different time series. The results revealed that the CsPLD genes are involved in the response to salinity stress and their expression levels enhance with increasing salinity stress period. The outcomes of this research will be useful for future molecular works related to lipid signaling in oilseed plants.

In silico identification of prospective virulence factors associated with candidiasis in Meyerozyma guilliermondii strain SO from genome dataset

BackgroundMeyerozyma guilliermondii is a prospective yeast that has extensively contributed to the biotechnology sector. In 2015, M. guilliermondii strain SO which was isolated from spoiled orange has successfully been developed as an inducer-free expression system and attained a significant impact in producing industrially important recombinant proteins. The species possesses high similarity to Candida albicans which may cause candidiasis. The industrial-benefiting M. guilliermondii strain SO has been underexplored for its virulence status. Thus, this study aimed to document the potential virulence factors through the comprehensive in silico analysis of M. guilliermondii strain SO genome. This analysis demonstrated the molecular characterization which could distinguish the pathogenicity status of M. guilliermondii.ResultsThe genome data were generated from Illumina HiSeq 4000 sequencing platform and assembled into 51 scaffolds successfully accumulating a genome size of 10.63 Mbp. These enclosed 5,335 CDS genes and 5,349 protein sequences with 43.72% GC content. About 99.29% of them were annotated to public databases. Komagataella phaffii, Saccharomyces cerevisiae and the reference strain of M. guilliermondii (ATCC 6260) were used as the controls. They were compared with our in-house strain SO to identify the consensus domain or subdomain which could putatively be considered as virulence factors. Candida albicans was used as the pathogenic model. Hence, hidden Markov model against strain SO proteome had identified secreted aspartic proteases (SAP), phospholipase C (PLC) and phospholipase D (PLD) with an E-value of 2.4e−107, 9.5e−200 and 0.0e+00, respectively, in resemblance of C. albicans. The topology of the phylogenetic analysis indicated that these virulence factors in M. guilliermondii strain SO and C. albicans branched from the same node and clustered together as a clade, signifying their molecular relatedness and congeneric among these species, subsequently proposing the virulence status of M. guilliermondii.ConclusionThe SAP, PLC and PLD genes’ features that were significant in expressing determinants of pathogenicity were successfully identified in M. guilliermondii strain SO genome dataset, thus concluding the virulency of this species. On account of this finding, the strategy of gene knockout through CRISPR-Cas9 or homologous recombination strategies is needed to engineer the feasible novel expression host system. Over and above, the genetically modified strain of M. guilliermondii allegedly may eradicate the risk of candidiasis infection.

Comprehensive in silico analysis of Phospholipase D gene family in economically important orchids

• A total of 25 putative PLD proteins from the genomes of Phalaenopsis equestris (7) , Dendrobium catenatum (9) and Apostasia shenzhenica (9) were identified. • These proteins are comprised of two HKD domains and two additional domains; C2 and PXPH domains at N-terminal region. • The multiple sequence alignment, domain and motifs analysis revealed their conserved nature and homology structure simulation and Ramachadran plot analysis revealed the abundance of beta sheets in tertiary structures. • The presence of Root-specific element, DOF proteins binding domain, scaffold or matrix attachment region, pollen-specific, dehydration-responsive, light-responsive, gibberellin-responsive, ABA-responsive elements and Meja-responsive upstream cis- regulatory promoter elements indicated their role in various physiological processes. Phospholipase D (PLD) plays a fundamental role in various aspects of plant growth, development and stress responses. In present study, 25 putative PLD proteins from the genomes of Phalaenopsis equestris (7) , Dendrobium catenatum (9) and Apostasia shenzhenica (9) were identified on the basis of presence of two HKD domains and two additional domains; C2 and PXPH domains at N-terminal region. These proteins were clustered into five sub-groups (α, β, δ, ζ and φ) along with their orthologs in Arabidopsis thaliana and Oryza sativa upon phylogenetic analysis. Gene characterization divulged that most of the PePLD, DcPLD and AsPLD genes within the same sub-group exhibited similar exon-intron organization and highly conserved motifs. RNA-seq based spatio-temporal studies indicated differential expression in various vegetative and reproductive tissues. PePLDα 1 , DcPLDα 1 and AsPLDα 1 were showing significant expression in all the tissues, while for other genes the expression was limited either in vegetative or reproductive tissues. Homology structure simulation revealed the abundance of random coils in secondary structures and beta sheets in tertiary structures. The presence of specific upstream cis- regulatory promoter elements emphasized the role of PLD s in various physiological processes. This report is the first genome-wide characterization of the PLD gene family in orchids and shall provide new insight into the functional characterization of these genes.

Functional characterization of transcription factor WIN1 genes associated with lipid biosynthesis and stress tolerance in soybean (Glycine max)

Transcription factor WAX INDUCER1 (WIN1) plays an important role in wax and cutin biosynthesis in plants. In this study, six GmWIN1 genes were identified from soybean ( Glycine max ), with GmWIN1–5 having high expression in flowers and seeds, particularly under cold stress. GmWIN1–5′s functions in lipid/oil biosynthesis and stress tolerance were characterized by its overexpression in soybean hairy roots and tobacco ( Nicotiana benthamiana ) plants. Under cold stress, GmWIN1–5 overexpression ( GmWIN1–5- OE) resulted in large oil enhancement and high level of phospholipids involved in cell membranes formation. Malondialdehyde content and elcetronlyte leakage in GmWIN1–5-OE plants was smaller than that in the wild-type control under cold stress, indicating that GmWIN1–5 promote plant stress tolerance. Moreover, GmWIN1–5-OE also led to upregulated expression of numerous genes involved in de novo fatty acid biosynthesis and triacylglycerol (TAG) production, including genes encoding phospholipase D ( PLD ), biotin carboxyl carrier protein 1 ( BCCP1 ), lysophosphatidic acid acyltransferase 5 ( LPAT5 ), and diacylglycerol acyltransferase 2 ( DGAT2 ). GmWIN1–5 directly bound to the promoter region of PLD gene. Overall, the results revealed that GmWIN1–5 can upregulate the biosynthesis and accumulation of lipids/oils, especially unsaturated fatty acids and phospholipids benefiting membrane integrity, and ultimately confer plants with high stress tolerance. • Six GmWIN1 members with structural integrity are identified in soybean ( Glycine max ). • Of the six GmWIN1s, GmWIN1-5 expression level was the highest in flower and seed tissues. • Overexpression of GmWiN1-5 in soybean hair roots increased total lipids and cold stress tolerance. • Ectopic expression of GmWIN1-5 in tobacco plant boosted oil content and stress tolerance. • GmWIN1-5 promoted expression of several lipid synthesis-related genes, particularly PLD.

Characterization and Expression of Phospholipase D Putatively Involved in Colletotrichummusae Disease Development of Postharvest Banana Fruit

Phospholipase D (PLD) in plants plays an important role in growth, development, and stress response. The effect of hexanal on PLD in banana fruit responding to Colletotrichum musae infection remains poorly understood. In this study, four putative PLD genes, named as MaPLD1, MaPLD2, MaPLD3, and MaPLD4 were identified from banana fruit. The four MaPLDs can be classified into three of the seven known PLD families according to sequence characterization. Their deduced amino acid sequences displayed homology of PLDs from other plant species. Furthermore, the specific expression analysis of PLD genes in banana fruit in response to infection in C. musae was studied and the response relationship between PLD family members and banana fruit under anthracnose stress was clarified. Changes in both the activity of PLD and PLC, and the connection between hexanal and phospholipases in the banana fruit C. musae infection were compared. The results showed that the incidence of disease in banana inoculated with C. musae was dramatically increased after 6 days of storage, the activation of PLD and PLC in infected anthracnose fruit before disease development, and that this activation was inhibited by hexanal treatment, which suggested that both enzymes play a protective role in banana fruit to cope with C. musae infection and the participation of hexanal in their regulation. Of the four MaPLD genes, the anthracnose had a stronger effect on MaPLD1 and MaPLD4. These data demonstrated that hexanal treatment could enhance fruit disease resistance to C. musae, and that PLD could take part in the disease defensive system of harvested banana fruit to C. musae by modulating the metabolism of cell membrane lipids, and thus suppress disease development in C. musae -inoculated banana during storage.

Genome-wide in silico identification of phospholipase D (PLD) gene family from Corchorus capsularis and Corchorus olitorius: reveals their responses to plant stress

BackgroundPlant grows in nature facing various types of abiotic stresses for their normal growth and development. During abiotic stress, plants evolve different types of mechanisms to survive in a hostile environment. Phospholipase D (PLD) plays important role in the regulation of diverse cellular processes including stress responses in plants. Member of PLD genes are well studied in different model plants; however, their functions in the jute are not clear yet. ResultIn the present study, a total of 12 and 11 PLD genes were identified in the genome of C. capsularis and C. olitorius, respectively. The presence of the two conserved HKD motifs in PLD genes except for CoPLDδ-2 in jute suggests their strong lipase activity. Twenty different motifs were found in the identified PLD genes, and PLD-β1, PLD-γ1, and all members of PLD-δ1 of both jute species contained the highest number of motifs. Phylogenetic analysis showed the close evolutionary relationship among the five groups of jute PLD proteins along with the PLD proteins from Arabidopsis. Tissue-specific expression pattern of PLDα1-2, PLD-α2, PLDβ1, PLDγ1, and PLDδ1 of two jute species suggested their involvement in plant growth and development. However, the expression pattern of PLDα1-2, PLDα1-3, PLD-α4, PLDδ1, and PLDδ3 indicated their association during waterlogging stress. In addition, PLD-α2, PLDβ1, and PLDδ2 seemed to be involved in drought stress as well as salinity stress. ConclusionThis genome-wide identification of jute PLD genes from C. capsularis and C. olitorius will help to further functional characterization of the PLD genes for developing stress-tolerant jute variety.

The Reduced Longitudinal Growth Induced by Overexpression of pPLAIIIγ Is Regulated by Genes Encoding Microtubule-Associated Proteins.

There are three subfamilies of patatin-related phospholipase A (pPLA) group of genes: pPLAI, pPLAII, and pPLAIII. Among the four members of pPLAIIIs (α, β, γ, δ), the overexpression of three isoforms (α, β, and δ) displayed distinct morphological growth patterns, in which the anisotropic cell expansion was disrupted. Here, the least studied pPLAIIIγ was characterized, and it was found that the overexpression of pPLAIIIγ in Arabidopsis resulted in longitudinally reduced cell expansion patterns, which are consistent with the general phenotype induced by pPLAIIIs overexpression. The microtubule-associated protein MAP18 was found to be enriched in a pPLAIIIδ overexpressing line in a previous study. This indicates that factors, such as microtubules and ethylene biosynthesis, are involved in determining the radial cell expansion patterns. Microtubules have long been recognized to possess functional key roles in the processes of plant cells, including cell division, growth, and development, whereas ethylene treatment was reported to induce the reorientation of microtubules. Thus, the possible links between the altered anisotropic cell expansion and microtubules were studied. Our analysis revealed changes in the transcriptional levels of microtubule-associated genes, as well as phospholipase D (PLD) genes, upon the overexpression of pPLAIIIγ. Overall, our results suggest that the longitudinally reduced cell expansion observed in pPLAIIIγ overexpression is driven by microtubules via transcriptional modulation of the PLD and MAP genes. The altered transcripts of the genes involved in ethylene-biosynthesis in pPLAIIIγOE further support the conclusion that the typical phenotype is derived from the link with microtubules.

Genome-Wide Analysis of the Phospholipase D Family in Five Cotton Species, and Potential Role of GhPLD2 in Fiber Development and Anther Dehiscence.

Phospholipase D (PLD) and its hydrolysis product phosphatidic acid play an important role in the regulation of several cellular processes, including root growth, pollen tube elongation, and microtubule reorganization. Here, we systematically identified and analyzed the membership, characterization, and evolutionary relationship of PLDs in five species of cotton. The results of the transcriptomic analysis suggested that the evaluated PLD genes showed high expression levels in anther tissue and during the fiber initiation and elongation periods. Quantitative real-time polymerase chain reaction showed differential expression of GhPLD genes in the anthers of photoperiod sensitive male sterility mutant 5 (psm5). Previous research on multiple stable quantitative trait loci also suggests the role of PLD genes in the fiber development. Further analyses showed that GhPLD2 protein is localized to the plasma membrane. The virus-induced gene silencing of GhPLD2 in cotton seedlings repressed its expression by 40–70%, which led to a reduction in reactive oxygen species (ROS) levels, 22% anther indehiscence, and disrupted fiber initiation and elongation. Thus, we inferred that GhPLD2 may promote ROS production, which, in turn, may regulate anther dehiscence and fiber development.

Genome-Wide Analysis and Expression Profiling of the Phospholipase D Gene Family in Solanum tuberosum.

Simple SummaryPhospholipase D is the most important kind of phospholipase in plants, which can specifically catalyze the hydrolysis of phosphodiester bonds at the end of phospholipid molecules to produce phospholipid acid and a free group. Phospholipase D is not only involved in maintaining the structural stability of plant cell membrane and the dynamic balance of lipid composition but also involved in a variety of physiological processes, such as plant growth and development and stress response, including stomatal closure, root elongation, cell senescence, high salt stress, cold stress, and drought stress. It plays a critical role in plant growth and development, as well as in hormone and stress responses. In this study, the basic information on phospholipase D gene family was comprehensively introduced, and the physical and chemical properties, systematic evolution, gene structure, conservative motif, chromosome location, gene replication, cis-acting element prediction, GO annotation, interspecific collinearity, and quantitative analysis of potato phospholipase D gene family were analyzed to gain a better understanding of potato phospholipase D gene family. Through the analysis of potato phospholipase D gene family, our results lay the foundation for further understanding of the function of phospholipase gene family in higher plants.Phospholipase D (PLD) is the most important phospholipid hydrolase in plants, which can hydrolyze phospholipids into phosphatidic acid (PA) and choline. When plants encounter low temperature, drought and high salt stress, phospholipase D and its products play an important role in regulating plant growth and development and coping with stress. In this study, 16 members of StPLD gene family were identified in potato genome, which were distributed in α, β, δ, and ζ subfamilies, and their expression patterns under salt, high temperature, drought, and ABA stress were detected by qRT-PCR method. Gene expression analysis showed that the expression of StPLD genes in potato was upregulated and downregulated to varying degrees under the four stresses, indicating that the PLD gene family is involved in the interaction of potato plant hormones and abiotic stress signals. Chromosome distribution showed that StPLD gene was unevenly distributed on 8 chromosomes, and only one pair of tandem repeat genes was found. All StPLD promoters contain hormone and stress-related cis-regulatory elements to respond to different stresses. Structural analysis showed that StPLD genes in the same subgroup had a similar exon–intron structure. Our study provides a valuable reference for further research of the function and structure of PLD gene.

Identification, evolution, expression analysis of phospholipase D (PLD) gene family in tea (Camellia sinensis).

The online version contains supplementary material available at 10.1007/s12298-021-01007-0.

Genetic Characterization of Corynebacterium pseudotuberculosis Isolates in Egypt

Corynebacterium pseudotuberculosis is a small Gram-positive bacillus containing mycolic acid in the structure of the cell wall. The bacterium is responsible for Caseous Lymphadenitis (CLA) in small ruminants (sheep and goats). The bacteria are also responsible for Ulcerative Lymphangitis in equines. The disease causes great economic loss in the animal industry. This work aimed to check the ability of Quadruplex PCR (Q-PCR) for genotyping and identification of Egyptian isolates of C. pseudotuberculosis and sequence analysis of phospholipase D (PLD) gene of local isolates. Four of C. pseudotuberculosis local isolates previously biochemically identified were tested for narG gene (nitrate reductase gene). Both nitrate negative biovar (ovis) and nitrate positive biovar (equi) showed a positive result for 16S rRNA, rpoB and PLD genes of C. pseudotuberculosis species. The sequence analysis of our local isolates' PLD gene revealed minor changes in PLD proteins between ovine and equine strains compared with other published PLD sequences in GenBank. It was concluded that the Q-PCR method is able to differentiate between C. pseudotuberculosis equi and ovis biovars. Also, the sequence of PLD gene of local isolates representing the two biovars revealed some variation, which leads to an accurate diagnosis of C. pseudotuberculosis biovars and generates a mapping of immerged local isolates. Further, the PCR and sequence of these isolates provide rapid and accurate genotyping, especially with hyperimmune serum's unavailability.

Membrane lipid metabolism in relation to core browning during ambient storage of ‘Nanguo’ pears

Abstract ‘Nanguo’ pears are prone to core browning at ambient temperatures. To reveal the role of membrane lipid metabolism in the process of core browning, fruit were treated with n-butanol, a phospholipase D (PLD) inhibitor. Changes in phospholipids and fatty acids, as well as the activities and gene expression of key enzymes were analyzed. Membrane phenotype and ultrastructure were also documented. Phospholipid and fatty acid levels, the activities of PLD and lipoxygenase (LOX), and gene expression changed during fruit ripening. PLD activity and gene expression in n-butanol-treated fruit were lower than those in control fruit, and the symptoms of core browning appeared later and were less severe. Furthermore, the membrane ultrastructure of fruit treated with n-butanol was intact compared with that in control fruit. These results suggested that membrane lipid metabolism plays an important role in the core browning of harvested ‘Nanguo’ pears, and that n-butanol treatment can alleviate disorder development by regulating membrane lipid metabolism.

Characterization of a PLDζ2 Homology Gene from Developing Castor Bean Endosperm.

Castor oil contains approximately 90% ricinoleic acid (RA) which is stored mainly in the form of tri-ricinoleic acid containing triacylglycerols (TAG). Ricinoleate is synthesized from oleate (18:1n-9) esterified to the sn-2 position of phosphatidylcholine (PtdCho) catalyzed by oleoyl-12-hydroxylase. PtdCho-derived diacylglycerol (DAG) is an important substrate pool for TAG synthesis, and the interconversion between PtdCho and DAG has been shown to play a critical role in channeling hydroxy fatty acids (HFA) to TAG. Although phospholipase D (PLD) has been reported to catalyze the hydrolysis of PtdCho to produce phosphatidic acid which can then be converted to DAG, its potential functions in the channeling of RA from PtdCho to DAG and the assembly of RA on TAG is largely unknown. In the present study, 11 PLD genes were identified from the Castor Bean Genome Database. Gene expression analysis indicated that RcPLD9 is expressed at relatively high levels in developing seeds compared to other plant tissues. Sequence and phylogenetic analyses revealed that RcPLD9 is a homolog of Arabidopsis PLDζ2. Overexpression of RcPLD9 in the Arabidopsis CL7 line producing C18-HFA resulted in RA content reductions in the polar lipid fraction (mainly PtdCho) and mono-HFA-TAG, but increased RA content in di-HFA-TAG. Since part of RA in di-HFA-TAG is derived from HFA-DAG, the results indicated that RcPLD9 facilitates the channeling of RA from PtdCho to DAG for its assembly on TAG in developing seeds.

Methyl jasmonate alleviates chilling injury by regulating membrane lipid composition in green bell pepper

The effects of methyl jasmonate (MeJA) treatment on membrane lipid metabolism of green bell pepper during cold storage was studied. The results showed that the chilling injury (CI) index, electrolyte leakage, malondialdehyde (MDA) content, phospholipase D (PLD) activity and gene expression of green bell pepper treated with MeJA were lower than those of the control fruit. MeJA treatment was able to maintain a higher level of ascorbic acid (VC) content, higher levels of phosphatidylcholine (PC), phosphatidylethanolamine (PE), phosphatidylserine (PS) and lower levels of digalactosyldiacylglycerol (DGDG), phosphatidic acid (PA), digalactosyldiacylglycerol/monogalactosyldiacylglycerol (DGDG/MGDG) and PC/PE compared to control fruit. In addition, MeJA treatment can also increase the proline content of green bell pepper fruit. Therefore, the reason that MeJA can improve the cold resistance of green bell pepper fruit may be attributed by the decreased expression and activity of PLD gene. The degradation of membrane lipid was delayed, and the content of proline increased, thereby maintaining cell membrane stability.

Phospholipase D (PLD) Response to Water Stress in Citrus Roots and Leaves

The effects of drought stress on phospholipase D (PLD) gene expression and enzymatic activity were investigated in ‘Pineapple’ sweet orange. PLDs are tissue-specific, with overlapping functions, and in response to stress they may interact with ABA signaling. Tissue specificity for expression of PLDs and their regulation are unknown in citrus. To assess PLD response to water stress and gene expression/regulation in citrus, we subjected potted seedlings to increasing levels of soil drought. Evapotranspirational demand (ET) was estimated by measuring weight loss in pots, and water stress was further assessed by measuring ABA content. Three treatments were performed over a 3-week period: (a) Control treatment without drought stress (100% of daily ET); (b) mild water stress (50% ET); and (c) severe water stress (0% ET). ABA content increased during drought stress in both roots and leaves, being higher in leaves than in roots by the end of the experiment for any stress condition assayed. PLD enzymatic activity was monitored and expression of five PLD genes was studied. PLD activity increased linearly over time in response to increased soil drought and was around three times higher in roots than in leaves. PLD activation occurred initially in roots and then in leaves. PLD gene expression in response to stress soil drought differed between roots and leaves. These results show the potential of PLD as a suitable indicator of stress severity in citrus.

Role of Phospholipase D Inhibitor in Regulating Expression of Senescencerelated Phospholipase D gene in Postharvest Longan Fruit

Background: Phospholipase D (PLD)is closely related to browning and senescence of postharvest longan fruit. Objective: This study investigated the effects of 2-butanol (a PLD inhibitor) on the expression and regulation of PLD during storage of longan fruit at a low temperature. Methods: Senescence-related quality indices showed that the 2-butanol-treated fruit presented lower pericarp browning index, pulp breakdown index and total soluble solid value than the untreated fruit. Results: The fruit treated by 60 µL/L 2-butanol exhibited the strongest inhibition on senescence, which significantly delayed changes in weight, titratable acidity content, total soluble solid content and ascorbic acid content. This treatment maintained a high level of total phenolic content and caused significant inhibition on pericarp browning and pulp breakdown. Through ELISA method, 60 µL/L 2-butanol treatment also reduced PLD activity. Real-time RT-PCR (RT-qPCR) results showed that PLD mRNA expression level was inhibited by 60 µL/L 2-butanol within 15 days. Western-blotting results further confirmed the differential expression of PLD during storage, and a relatively higher expression for PLD protein was found in control compared to the 2-butanoltreated fruit during 15-d storage. Conclusion: These results provided a scientific basis and reference to further investigating postharvest longan quality maintenance by regulating the PLD gene expression.