Two maize homologs of mammalian proton-coupled folate transporter, ZmMFS_1-62 and ZmMFS_1-73, are essential to salt and drought tolerance.

Folates are essential to the maintenance of normal life activities in almost all organisms. Proton-coupled folate transporter (PCFT), belonging to the major facilitator superfamily, is one of the three major folate transporter types widely studied in mammals. However, information about plant PCFTs is limited. Here, a genome-wide identification of maize PCFTs was performed, and two PCFTs, ZmMFS_1-62 and ZmMFS_1-73, were functionally investigated. Both proteins contained the typical 12 transmembrane helixes with N- and C-termini located in the cytoplasm, and were localized in the plasma membrane. Molecular docking analysis indicated their binding activity with folates via hydrogen bonding. Interference with ZmMFS_1-62 and ZmMFS_1-73 in maize seedlings through virus-induced gene silencing disrupted folate homeostasis, mainly in the roots, and reduced tolerance to drought and salt stresses. Moreover, a molecular chaperone protein, ZmHSP20, was found to interact with ZmMFS_1-62 and ZmMFS_1-73, and interference with ZmHSP20 in maize seedlings also led to folate disruption and increased sensitivity to drought and salt stresses. Overall, this is the first report of functional identification of maize PCFTs, which play essential roles in salt and drought stress tolerance, thereby linking folate metabolism with abiotic stress responses in maize.

Integration of Metabolomics and Transcriptomics to Explore Dynamic Alterations in Fruit Color and Quality in 'Comte de Paris' Pineapples during Ripening Processes.

Pineapple color yellowing and quality promotion gradually manifest as pineapple fruit ripening progresses. To understand the molecular mechanism underlying yellowing in pineapples during ripening, coupled with alterations in fruit quality, comprehensive metabolome and transcriptome investigations were carried out. These investigations were conducted using pulp samples collected at three distinct stages of maturity: young fruit (YF), mature fruit (MF), and fully mature fruit (FMF). This study revealed a noteworthy increase in the levels of total phenols and flavones, coupled with a concurrent decline in lignin and total acid contents as the fruit transitioned from YF to FMF. Furthermore, the analysis yielded 167 differentially accumulated metabolites (DAMs) and 2194 differentially expressed genes (DEGs). Integration analysis based on DAMs and DEGs revealed that the biosynthesis of plant secondary metabolites, particularly the flavonol, flavonoid, and phenypropanoid pathways, plays a pivotal role in fruit yellowing. Additionally, RNA-seq analysis showed that structural genes, such as FLS, FNS, F3H, DFR, ANR, and GST, in the flavonoid biosynthetic pathway were upregulated, whereas the COMT, CCR, and CAD genes involved in lignin metabolism were downregulated as fruit ripening progressed. APX as well as PPO, and ACO genes related to the organic acid accumulations were upregulated and downregulated, respectively. Importantly, a comprehensive regulatory network encompassing genes that contribute to the metabolism of flavones, flavonols, lignin, and organic acids was proposed. This network sheds light on the intricate processes that underlie fruit yellowing and quality alterations. These findings enhance our understanding of the regulatory pathways governing pineapple ripening and offer valuable scientific insight into the molecular breeding of pineapples.

The dynamic changes of mango (Mangifera indica L.) epicuticular wax during fruit development and effect of epicuticular wax on Colletotrichum gloeosporioides invasion.

Mango fruits are susceptible to diseases, such as anthracnose, during fruit development, leading to yield reduction. Epicuticular wax is closely related to resistance of plants to pathogenic bacterial invasion. In this study, the effect of mango fruit epicuticular wax on the invasion of Colletotrichum gloeosporioides was investigated, followed by to understand the changes of wax chemical composition and crystal morphology during mango fruit development using GC-MS and SEM. Results showed that the epicuticular wax of mango fruits can prevent the invasion of C. gloeosporioides, and 'Renong' showed the strongest resistance to C. gloeosporioides. The wax content of four mango varieties first increased and then decreased from 40 days after full bloom (DAFB) to 120 DAFB. In addition, 95 compounds were detected in the epicuticular wax of the four mango varieties at five developmental periods, in which primary alcohols, terpenoids and esters were the main wax chemical composition. Furthermore, the surface wax structure of mango fruit changed dynamically during fruit development, and irregular platelet-like crystals were the main wax structure. The present study showed the changes of wax content, chemical composition and crystal morphology during mango fruit development, and the special terpenoids (squalene, farnesyl acetate and farnesol) and dense crystal structure in the epicuticular wax of 'Renong' fruit may be the main reason for its stronger resistance to C. gloeosporioides than other varieties. Therefore, these results provide a reference for the follow-up study of mango fruit epicuticular wax synthesis mechanism and breeding.

Transcriptomic and proteomic analyses of Mangifera indica in response to Xanthomonas critis pv. mangiferaeindicae.

Mango is an important tropical fruit with the reputation of "Tropical Fruit King." It is widely cultivated in tropical and subtropical regions. Mango bacterial leaf spot, which is caused by Xanthomonas critis pv. mangiferaeindicae (Xcm), poses a great threat to the development of mango planting industry. In this study, we used RNA sequencing and data-independent acquisition techniques to compare the transcriptome and proteome of the highly resistant cultivar "Renong No.1" (RN) and the highly susceptible cultivar "Keitt" (KT) in response to Xcm infection at different stages (0, 2, and 6 days). A total of 14,397 differentially expressed genes (DEGs) were identified in the transcriptome of the two varieties, and 4,400 and 8,926 genes were differentially expressed in RN and KT, respectively. Among them, 217 DEGs were related to plant hormone signaling pathway, and 202 were involved in the maintenance of cellular redox homeostasis. A total of 3,438 differentially expressed proteins (DEPs) were identified in the proteome of the two varieties. Exactly 1,542 and 1,700 DEPs were detected in RN and KT, respectively. In addition, 39 DEPs were related to plant hormone signaling pathway, whereas 68 were involved in the maintenance of cellular redox homeostasis. Through cross-validation of the two omics, 1,470 genes were found to be expressed in both groups, and a large number of glutathione metabolism-related genes, such as HSP26-A, G6PD4, and GPX2, were up-regulated in both omics. Peroxisome-related genes, such as LACS6, LACS9, PED1, GLO4, and HACL, were up-regulated or down-regulated in both omics. ABCB11, SAPK2, MYC2, TAG7, PYL1, and other genes related to indole-3-acetic acid and abscisic acid signal transduction and plant-pathogen interaction were up-regulated or down-regulated in both omics. We also used weighted gene co-expression network analysis to combine physiological and biochemical data (superoxide dismutase and catalase activity changes) with transcriptome and proteome data and finally identified three hub genes/proteins (SAG113, SRK2A, and ABCB1) that play an important role in plant hormone signal transduction. This work was the first study of gene/protein changes in resistant and susceptible mango varieties, and its results improved our understanding of the molecular mechanism of mango resistance to Xcm.

Low temperature storage alleviates internal browning of 'Comte de Paris' winter pineapple fruit by reducing phospholipid degradation, phosphatidic acid accumulation and membrane lipid peroxidation processes.

To uncover the mechanism underlying membrane lipid metabolism of low temperature induced internal browning tolerance in pineapple, membrane phospholipid alterations of harvested 'Comte de Paris' winter pineapple fruit stored at either 10 °C or 25 °C was investigated. Fruit stored at 10 °C developed low levels of internal browning as compared to fruit stored at 25 °C and was associated with high contents of phosphatidylcholine, phosphatidylglycerol, phosphatidylinositol, phosphatidylserine, and phosphatidylethanolamine, and low levels of phosphatidic acid. Storage at 10 °C down-regulated the expression levels of phospholipase As. Fruit stored at 10 °C also exhibited high ratio of unsaturated fatty acid to saturated fatty acid and index of unsaturated fatty acid level. These findings suggest that maintenance phospholipid abundance, reduction in phosphatidic acid accumulation and membrane lipid peroxidation may have contributed to the enhanced internal browning tolerance in 'Comte de Paris' winter pineapple fruit at low temperature storage.

The class III peroxidase gene family is involved in ascorbic acid induced delay of internal browning in pineapple.

Excessive production of reactive oxygen species (ROS) leads to potential toxicity in an organism. Class III peroxidases (PRXs) play an important role in maintaining ROS homeostasis in plants. Internal browning (IB) limits industrial development of pineapple, which is the third most important fruit trade in the world. IB is mainly caused by ROS, and the mechanism underlying IB is still unknown from the perspective of ROS. Here, we soaked pineapples in ascorbic acid after harvest and before storage to decrease excessive ROS and polyphenol oxidase (PPO) activity, ultimately restraining the spread and deterioration of IB. Using phylogenetic analysis; we identified 78 pineapple PRX genes (AcPRXs) and divided them into five subgroups. Gene structure analysis indicated that the exon numbers ranged from 2 to 14, and conserved motif analysis verified that all of the AcPRXs identified here have standard peroxidase domains. Analysis of duplication events suggested that tandem and segmental duplication events may have played equal and important roles in expanding the AcPRX family. Comprehensive transcriptomic analysis uncovered that AcPRXs may play an important role in negatively regulating the occurrence of IB. In summary, we found that ROS scavenging delayed IB occurrence. The results of characterized AcPRX family revealed that AcPRXs family responded to growth and development, and negatively regulated to IB occurrence in storage stage. This research provides potential target genes for future in-depth analysis of the molecular mechanisms underlying IB and contributes to develop IB-resistant pineapple varieties.

Novel Insight into the Relationship between Metabolic Profile and Fatty Acid Accumulation Altering Cellular Lipid Content in Pineapple Fruits at Different Stages of Maturity.

Pineapple fruits are usually harvested at different stages of maturity, based on consumer demands. The stage of maturity significantly affects the storage tolerance due to alterations in the cellular lipid homeostasis in the fruits. The characteristic abundance of metabolites and fatty acids (FAs) can provide vital information giving insight into the cellular lipid changes that occur during the ripening process in the fruits. Here, liquid chromatography-tandem mass spectrometry, largely based on the analysis of widely targeted metabolomics, was applied to evaluate the differences in the metabolites among the pineapple at three different stages of maturity namely, pineapples at the young fruit (YF), mature fruit (MF), and fully mature fruit (FMF) stages. In this study, 466 metabolites were annotated and identified. Among these, 59 lipids, including the glyceride esters, fatty acids and conjugates, and lysophospholipids (LPLs) were characterized. Notably, the LPLs were down-regulated in their relative abundance in the MF compared with the YF, and subsequently they remained almost stable in the FMF stage. The FA profiling results revealed the presence of certain unsaturated fatty acids (UFAs); besides, the total monounsaturated fatty acid (MUFA) to saturated fatty acid (SFA) ratio, as well as the polyunsaturated fatty acids (PUFA) to SFA ratio, showed noticeable decrease during the ripening process. The differential accumulation patterns of the LPLs, MUFAs, PUFAs, and SFAs imply that the lipid degradation and peroxidation take place in the pineapple fruits from the YF to MF and YF to FMF stages, respectively. The present study provides new insights into the alterations in the cellular lipid metabolism underlying the metabolite profiles and accumulation of FAs in pineapple fruits during ripening.

Optimization extraction, characterization and anticancer activities of polysaccharides from mango pomace.

Response surface methodology was used to optimize the extraction conditions for ultrasonic-assisted extraction of polysaccharides from mango pomace. The Optimum extraction conditions consisted of extraction temperature of 74 °C, ultrasonic power of 170 W, extraction time of 100 min, and raw material-to-water ratio of 1:40 g/mL. Under these conditions, the extraction yield was 3.71 ±â€¯0.07%. Three novel polysaccharide fractions, MG-1, MG-2 and MG-3 were purified from the crude polysaccharides by using DEAE-52 cellulose and Sephadex G-100 column chromatography. The molecular weight and monosaccharide composition of polysaccharide fractions (MPFs) were analyzed by high performance liquid gel permeation chromatography (HPGPC) and HPLC analysis, respectively. The characterizations of MPFs were conducted with FT-IR, 1H NMR and SEM. Furthermore, the anticancer activities of the polysaccharide fractions were also investigated in vitro. Results showed that MG-1, MG-2 and MG-3 exhibited significant anticancer activities against HepG2, MCF-7, A549, HeLa, A2780, HCT-116 and BGC-823 cells in a dose-dependent manner. MPFs were also showed to promote apoptosis as seen in the nuclear morphological examination study using calcein acetyl methoxy methyl easter (calcein-AM) and propidium iodide (PI) staining. This research could serve as a theoretical reference for the efficient utilization of MPFs in biomedical and functional food.

Profiling of volatile fragrant components in a mini-core collection of mango germplasms from seven countries.

Aroma is important in assessing the quality of fresh fruit and their processed products, and could provide good indicators for the development of local cultivars in the mango industry. In this study, the volatile diversity of 25 mango cultivars from China, America, Thailand, India, Cuba, Indonesia, and the Philippines was investigated. The volatile compositions, their relative contents, and the intervarietal differences were detected with headspace solid phase microextraction tandem gas chromatography-mass spectrometer methods. The similarities were also evaluated with a cluster analysis and correlation analysis of the volatiles. The differences in mango volatiles in different districts are also discussed. Our results show significant differences in the volatile compositions and their relative contents among the individual cultivars and regions. In total, 127 volatiles were found in all the cultivars, belonging to various chemical classes. The highest and lowest qualitative abundances of volatiles were detected in 'Zihua' and 'Mallika' cultivars, respectively. Based on the cumulative occurrence of members of the classes of volatiles, the cultivars were grouped into monoterpenes (16 cultivars), proportion and balanced (eight cultivars), and nonterpene groups (one cultivars). Terpene hydrocarbons were the major volatiles in these cultivars, with terpinolene, 3-carene, caryophyllene and α-Pinene the dominant components depending on the cultivars. Monoterpenes, some of the primary volatile components, were the most abundant aroma compounds, whereas aldehydes were the least abundant in the mango pulp. ß-Myrcene, a major terpene, accounted for 58.93% of the total flavor volatile compounds in 'Xiaofei' (Philippens). γ-Octanoic lactone was the only ester in the total flavor volatile compounds, with its highest concentration in 'Guiya' (China). Hexamethyl cyclotrisiloxane was the most abundant volatile compound in 'Magovar' (India), accounting for 46.66% of the total flavor volatiles. A typical aldehydic aroma 2,6-di-tert-butyl-4-sec-butylphenol, was detected in 'Gleck'. A highly significant positive correlation was detected between Alc and K, Alk and Nt, O and L. Cultivars originating from America, Thailand, Cuba, India, Indonesia and the Philippines were more similar to each other than to those from China. This study provides a high-value dataset for use in development of health care products, diversified mango breeding, and local extension of mango cultivars.

Transcriptome and proteomic analysis of mango (Mangifera indica Linn) fruits.

Here we used Illumina RNA-seq technology for transcriptome sequencing of a mixed fruit sample from 'Zill' mango (Mangifera indica Linn) fruit pericarp and pulp during the development and ripening stages. RNA-seq generated 68,419,722 sequence reads that were assembled into 54,207 transcripts with a mean length of 858bp, including 26,413 clusters and 27,794 singletons. A total of 42,515(78.43%) transcripts were annotated using public protein databases, with a cut-off E-value above 10(-5), of which 35,198 and 14,619 transcripts were assigned to gene ontology terms and clusters of orthologous groups respectively. Functional annotation against the Kyoto Encyclopedia of Genes and Genomes database identified 23,741(43.79%) transcripts which were mapped to 128 pathways. These pathways revealed many previously unknown transcripts. We also applied mass spectrometry-based transcriptome data to characterize the proteome of ripe fruit. LC-MS/MS analysis of the mango fruit proteome was using tandem mass spectrometry (MS/MS) in an LTQ Orbitrap Velos (Thermo) coupled online to the HPLC. This approach enabled the identification of 7536 peptides that matched 2754 proteins. Our study provides a comprehensive sequence for a systemic view of transcriptome during mango fruit development and the most comprehensive fruit proteome to date, which are useful for further genomics research and proteomic studies. BIOLOGICAL SIGNIFICANCE: Our study provides a comprehensive sequence for a systemic view of both the transcriptome and proteome of mango fruit, and a valuable reference for further research on gene expression and protein identification. This article is part of a Special Issue entitled: Proteomics of non-model organisms.

Isolation and characterization of NBS-LRR resistance gene analogues from mango.

The nucleotide-binding site (NBS)-leucine-rich repeat (LRR) gene family is a class of R genes in plants. NBS genes play a very important role in disease defence. To further study the variation and homology of mango NBS-LRR genes, 16 resistance gene analogues (RGAs) (GenBank accession number HM446507-22) were isolated from the polymerase chain reaction fragments and sequenced by using two degenerate primer sets. The total nucleotide diversity index Pi was 0.362, and 236 variation sites were found among 16 RGAs. The degree of homology between the RGAs varied from 44.4% to 98.5%. Sixteen RGAs could be translated into amino sequences. The high level of this homology in the protein sequences of the P-loop and kinase-2 of the NBS domain between the RGAs isolated in this study and previously characterized R genes indicated that these cloned sequences belonged to the NBS-LRR gene family. Moreover, these 16 RGAs could be classified into the non-TIR-NBS-LRR gene family because only tryptophan (W) could be claimed as the final residual of the kinase-2 domain of all RGAs isolated here. From our results, we concluded that our mango NBS-LRR genes possessed a high level of variation from the mango genome, which may allow mango to recognize many different pathogenic virulence factors.

Isolation and structural characterization of a novel polysaccharide prepared from Arca subcrenata Lischke.

A water-soluble polysaccharide was isolated from Arca subcrenata Lischke (named ASLP) by hot-water extraction, anion-exchange, and gel-permeation chromatography. The average molecular weight of ASLP was estimated to be 3500 Da. The structural characterization of ASLP was performed by sugar composition analysis, methylation analysis, and partial acidic hydrolysis. Further analysis of ASLP was carried out by UV, FT-IR and NMR spectroscopies (1D, COSY, and HSQC, respectively). Our data suggests that ASLP is an alpha-(1-->4)-D-glucan, with an alpha-(1-->6)-D-glucan at the C-6 position every fourth residue along the main chain. The branch chain has three glucose residues. The possible structure, determined on the basis of structural analyses results, was also determined. Preliminary in vitro tests revealed that ASLP can stimulate mouse spleen lymphocyte proliferation and its branches are extremely important for its immunological activity.