Comparative proteome analysis of rubber latex serum from pathogenic fungi tolerant and susceptible rubber tree (Hevea brasiliensis)

Havanapan, Phattara‐orn; Bourchookarn, Apichai; Ketterman, Albert J.; Krittanai, Chartchai

doi:10.1016/j.jprot.2015.10.014

Cited by 20 publications

(13 citation statements)

References 41 publications

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“…A total of 150 μg of protein from the re-suspended protein extracts were passively rehydrated overnight on 7 cm non-linear IPG strips (pH 3–10) and then focused to 10,000 VH using the Protean IEF cell (BioRad Ltd, Hemel Hempsted,UK) as described previously [40] . The use of 7 cm strips is consistent with previous studies into the plant proteome [41] , [42] , [43] . Equilibration of each IPG strip was conducted for 12 min in 2.5 ml equilibration buffer (50 mM Tris-HCl pH 8.8, 6 M urea, 30% glycerol v/v and 2% SDS w/v), with the presence of DTT (Melford, Ipswich, U.K.) at 10 mg/ml followed by a second equilibration with IAA (iodoacetamide) (Sigma, Gillingham, U.K.) at 25 mg/ml.…”

Section: Methodssupporting

confidence: 86%

The effects of PPO activity on the proteome of ingested red clover and implications for improving the nutrition of grazing cattle

Hart

Onime

Davies

et al. 2016

Journal of Proteomics

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Increasing the rumen-stable protein content of feed would lead to improved nitrogen utilisation in cattle, and less nitrogenous waste. Red clover (Trifolium pratense L.) is a high protein ruminant feed containing high polyphenol oxidase (PPO) activity. PPO mediated protein-quinone binding has been linked to protecting plant proteins from proteolysis. To explore the mechanism underlying the effect of PPO on protein protection in fresh forage feeds, proteomic components of feed down-boli produced from wild-type red clover and a low PPO mutant, at point of ingestion and after 4 h in vitro incubation with rumen inoculum were analysed. Significant differences in proteomic profiles between wild-type and mutant red clover were determined after 4 h incubation, with over 50% less spots in mutant than wild-type proteomes, indicating decreased proteolysis in the latter. Protein identifications revealed preferentially retained proteins localised within the chloroplast, suggesting that PPO mediated protection in the wild-type operates due to the proximity of target proteins to the enzyme and substrates, either diffusing into this compartment from the vacuole or are present in the chloroplast. This increased understanding of protein targets of PPO indicates that wider exploitation of the trait could contribute to increased protein use efficiency in grazing cattle.Biological significanceOne of the main challenges for sustainable livestock farming is improving capture of dietary nitrogen by ruminants. Typically up to 70% of ingested protein-N is excreted representing a loss of productivity potential and a serious environmental problem in terms of nitrogenous pollution of lands and water. Identification of key characteristics of rumen-protected protein will deliver target traits for selection in forage breeding programmes. The chloroplastic enzyme PPO catalyzes the oxidation of phenols to quinones, which react with protein. Little is currently known about the intracellular protein targets of the products of PPO activity or the mechanism underlying protein complexing, including whether there is any specificity to the reaction. Here we have determined significant differences in the proteomes of freshly ingested down boli corresponding to the presence or absence of active PPO. These results show that in the presence of PPO the forage protein is less amenable to proteolysis and provide the novel information that the protected proteins are putatively chloroplastically located. These data also contribute to a growing evidence base that a chloroplastic PPO substrate exists in red clover in addition to the currently known vacuolar substrates.

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Section: Methodssupporting

confidence: 86%

The effects of PPO activity on the proteome of ingested red clover and implications for improving the nutrition of grazing cattle

Hart

Onime

Davies

et al. 2016

Journal of Proteomics

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“…H . brasiliensis clone RRIM 600, a secondary clone originally developed from clones Tijr 1 and PB 86, is the most widely planted clone in Malaysia and Thailand1314. RRIM 600 is categorized as a medium yielding clone that produces ~1,350 kg of latex per hectare per year in estates over 20 years old.…”

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confidence: 99%

The rubber tree genome shows expansion of gene family associated with rubber biosynthesis

Lau

Makita

Kawashima

et al. 2016

Sci Rep

110

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Hevea brasiliensis Muell. Arg, a member of the family Euphorbiaceae, is the sole natural resource exploited for commercial production of high-quality natural rubber. The properties of natural rubber latex are almost irreplaceable by synthetic counterparts for many industrial applications. A paucity of knowledge on the molecular mechanisms of rubber biosynthesis in high yield traits still persists. Here we report the comprehensive genome-wide analysis of the widely planted H. brasiliensis clone, RRIM 600. The genome was assembled based on ~155-fold combined coverage with Illumina and PacBio sequence data and has a total length of 1.55 Gb with 72.5% comprising repetitive DNA sequences. A total of 84,440 high-confidence protein-coding genes were predicted. Comparative genomic analysis revealed strong synteny between H. brasiliensis and other Euphorbiaceae genomes. Our data suggest that H. brasiliensis’s capacity to produce high levels of latex can be attributed to the expansion of rubber biosynthesis-related genes in its genome and the high expression of these genes in latex. Using cap analysis gene expression data, we illustrate the tissue-specific transcription profiles of rubber biosynthesis-related genes, revealing alternative means of transcriptional regulation. Our study adds to the understanding of H. brasiliensis biology and provides valuable genomic resources for future agronomic-related improvement of the rubber tree.

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“…10 The proteins constituents found in FTIR investigation could be responsible for the antioxidant properties of latex C-serum found in this work. Havanapan et al (2015) 38 investigated the protein contents of latex serum from H. brasiliensis by proteomic analysis and beta-1,3-glucanase, chitinase and lectin were isolated. These proteins are associated with plant defenses against microbes such as fungus.…”

Section: Discussionmentioning

confidence: 99%

Free-radical Scavenging properties and Cytotoxic Activity Evaluation of Latex C-serum from Hevea brasiliensis RRIM 600

Kerche

Cavalcante

Danna

et al. 2016

FRA

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Background: There are few studies exploring the antioxidant properties of the latex C-serum of the Hevea brasiliensis latex and their possible use for therapeutic application. Objective: This study investigated the in vitro antioxidant activity of the latex C-serum of H. brasiliensis and assessed the cytotoxicity of this fraction in two human cell lines derived from normal tissue (MRC-5 and CCD 1059sk. Chemical characterization was also assessed. Methods: Latex from H. brasiliensis was centrifuged and the C-serum phase was tested for in vitro free radical scavenging assays, such as hydroxyl and nitric oxide radicals, hydrogen peroxide and total antioxidant capacity by the phosphomolibdate method. Latex C-serum was also investigated for cytotoxic properties by the MTT test. Latex C-serum was chemical characterized by Infrared Spectroscopy Fourier Transform (FTIR). Results: Latex C-serum effectively scavenged hydroxyl and nitric oxide radicals and hydrogen peroxide and presented potent antioxidant activity. The results were compared to ascorbic acid, a standard antioxidant. The cytotoxicity evaluation of latex C-serum demonstrated cell line dependency and for CCD 1059sk cells derived from skin fibroblasts, latex C-serum was not cytotoxic. FTIR showed the presence of proteins in latex C-serum. Conclusion: Latex C-serum from Hevea brasiliensis presented strong antioxidant potential and these results openthe possibility of further studies aiming the use of latex C-serum as treatment for skin diseases related to reactive oxygenspecies or skin preventive pharmacological formulations.

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Comparative proteome analysis of rubber latex serum from pathogenic fungi tolerant and susceptible rubber tree (Hevea brasiliensis)

Cited by 20 publications

References 41 publications

The effects of PPO activity on the proteome of ingested red clover and implications for improving the nutrition of grazing cattle

The effects of PPO activity on the proteome of ingested red clover and implications for improving the nutrition of grazing cattle

The rubber tree genome shows expansion of gene family associated with rubber biosynthesis

Free-radical Scavenging properties and Cytotoxic Activity Evaluation of Latex C-serum from Hevea brasiliensis RRIM 600

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