Glutamine regulates the human epithelial intestinal HCT-8 cell proteome under apoptotic conditions

Glutamine plays a key role in the metabolism of rapidly dividing cells, including enterocytes and lymphocytes, which may contribute to its beneficial clinical effects. Gut mucosal homeostasis is achieved through a balance between cell proliferation and apoptosis. In T cells, glutamine up-regulates antiapoptotic proteins and down-regulates proapoptotic proteins. In gut mucosa, glutamine prevents apoptosis in rat epithelial cell lines, whereas glutamine starvation induces apoptosis through caspase activation. Finally glutamine specifically prevents tumor necrosis factor-alpha-related apoptosis in the human intestinal cell line HT-29. Comparative functional proteomics enables the characterization of each differentially expressed protein in intestinal cells in response to modifications of nutritional environment. The influence of glutamine on intestinal proteome expression in apoptotic conditions has not been studied and evaluated. This comparative proteomics study was performed in the human epithelial intestinal cell line HCT-8 under experimental apoptotic conditions to investigate the influence of glutamine on protein expression during apoptosis. The pharmaconutritional effects of glutamine were determined under 2 mm (physiological concentration) and 10 mm (pharmaconutritional concentration) conditions. About 1,800 protein spots were revealed in both conditions. Comparative assessments indicated that 28 proteins were differentially expressed significantly (i.e. at least 2-fold modulated and Student's t test with p 相似文献   

Proteomic analysis of glutamine-treated human intestinal epithelial HCT-8 cells under basal and inflammatory conditions

Glutamine (Gln) promotes intestinal growth and maintains gut structure and function, especially in situations of injury and during inflammation. Several mechanisms could contribute to Gln protective effects on gut. Proteomics enable us to characterize differentially expressed proteins in tissues in response to modifications of the biological or nutritional environment. Gln effects on the human intestinal epithelial HCT-8 cell line proteome were assessed under basal and proinflammatory conditions. The 2-DE gels were obtained and compared. Proteins were identified by MS and using databases. About 1200 spots were detected in both 2- and 10-mM Gln concentrations. Under basal conditions, 24 proteins were differentially expressed in response to Gln. Half of these proteins were implicated in protein biosynthesis or proteolysis and 20% in membrane trafficking. Under proinflammatory conditions, 27 proteins were up- or down-regulated by Gln 10 mM. From these proteins, 40% were involved in protein biosynthesis or proteolysis, 16% in membrane trafficking, 8% in cell cycle and apoptosis mechanisms and 8% in nucleic acid metabolism. This study provides the first holistic picture of proteome modulation by Gln in a human enterocytic cell line under basal and proinflammatory conditions, and supports further evaluation of nutritional modulation of intestinal proteome in humans.     

Effect of cocoa pod age on egg distribution and egg parasitism of the cocoa pod borer in Malaysia

The effects of pod age on oviposition, and on egg and larval survival of the cocoa pod borer (CPB), Conopomorpha cramerella (Snellen), were investigated. CPB were allowed to oviposit on a batch of 50 pods for a period of 7 days over several exposure dates. Numbers of CPB eggs/pod and survival to larval emergence from the pods were recorded. Pods were harvested when ripe and analyzed for larval survival within the pod. The oviposition pattern varied with pod age. Older pods less than 7 weeks before ripening (WBR) were preferred, suggesting greater nutritional value or chemical attraction of the older pods. The proportion of eggs parasitized by Trichogrammtoidea bactrae fumata Nagaraja (TBF), the number of eggs lost and the proportion surviving to larval emergence from the pod were independent of pod age. Egg parasitism was low, implying that TBF was not an effective natural enemy. Mass-releases of TBF should be intensified during cropping periods with higher proportions of susceptible pods (<10 WBR). Survival of larvae in the presclerotic layer (preSCL) was high and independent of pod age. Larval mortality at the SCL (sclerotic layer) was dependent on pod age and was high in older pods. Larval density influenced the survival of larvae inside the pod and was dependent on pod age. Larval mortality associated with SCL hardness and thickness is one of the criteria for the selection of CPB resistant cocoa clones.     

Proteomics Analysis of Soybean Seedlings under Short-Term Water Deficit

Soybeans are one of the most important grain crops worldwide. Water deficit, which seriously affects the yield and quality of soybeans, is the main abiotic stress factor in soybean production. As a follow-up study, the droughttolerant soybean variant Heinong 44 was analyzed via proteome analysis. Soybean was exposed to water deficit for 0, 8, and 24 h, and protein samples were extracted for detection of differentially expressed proteins. Protein sequencing of leaf tissues under water stress yielded a total of 549 differentially expressed proteins: 75 and 320 upregulated proteins as well as 70 and 84 downregulated proteins were obtained after 8 and 24 h of water deficit, respectively. Gene Ontology analysis revealed that most of the differentially expressed proteins (DEPs) were involved in catalytic activity, molecular function, and metabolic processes, whereas some of them were involved in photosynthesis, carbon metabolism, and energy metabolism. We also identified some differentially expressed proteins that may be involved in the regulation of water deficit response. Our study provides a theoretical basis for the breeding of drought-resistant soybean varieties.     

The role of the pod in seed development: strategies for manipulating yield

Pods play a key role in encapsulating the developing seeds and protecting them from pests and pathogens. In addition to this protective function, it has been shown that the photosynthetically active pod wall contributes assimilates and nutrients to fuel seed growth. Recent work has revealed that signals originating from the pod may also act to coordinate grain filling and regulate the reallocation of reserves from damaged seeds to those that have retained viability. In this review we consider the evidence that pods can regulate seed growth and maturation, particularly in members of the Brassicaceae family, and explore how the timing and duration of pod development might be manipulated to enhance either the quantity of crop yield or its nutritional properties.     

Dissecting the proteome of pea mature seeds reveals the phenotypic plasticity of seed protein composition

Pea (Pisum sativum L.) is the most cultivated European pulse crop and the pea seeds mainly serve as a protein source for monogastric animals. Because the seed protein composition impacts on seed nutritional value, we aimed at identifying the determinants of its variability. This paper presents the first pea mature seed proteome reference map, which includes 156 identified proteins (http://www.inra.fr/legumbase/peaseedmap/). This map provides a fine dissection of the pea seed storage protein composition revealing a large diversity of storage proteins resulting both from gene diversity and post‐translational processing. It gives new insights into the pea storage protein processing (especially 7S globulins) as a possible adaptation towards progressive mobilization of the proteins during germination. The nonstorage seed proteome revealed the presence of proteins involved in seed defense together with proteins preparing germination. The plasticity of the seed proteome was revealed for seeds produced in three successive years of cultivation, and 30% of the spots were affected by environmental variations. This work pinpoints seed proteins most affected by environment, highlighting new targets to stabilize storage protein composition that should be further analyzed.     

Quantitative proteomics to study the response of wheat to contrasting fertilisation regimes

Negative environmental impacts from mineral fertilisers and pesticides used in conventional cropping have raised concern over the sustainability of arable crop production. Organic cropping uses alternatives that avoid many of these negative environmental effects; however, crop yields can be significantly reduced, possibly due to a lower proportion of plant-available nutrients. To gain insights into the molecular effects of organic compared to conventional cropping systems on plant utilisation of nutrients, we used proteomics to analyse winter wheat (Triticum aestivum). Our aim was to investigate the effects of contrasting fertility management and crop protection regimes in organic and conventional cropping systems on the wheat flag leaf proteome and the association between the proteome and physiological traits. Wheat flag leaves were flash-frozen, lyophilised and milled prior to protein extraction (TCA/acetone) and analysed using 2D gel electrophoresis and MALDI-TOF MS. The abundance of 111 protein spots varied significantly between fertilisation regimes. Flag leaf N and P composition were significant drivers of differences in protein spot abundance, including major proteins involved in nitrogen remobilisation, photosynthesis, metabolism and stress response. These results indicate that molecular-based mechanisms are involved in the effect of contrasting cropping systems on nutrient utilisation and wheat grain yield. Using a functional genomics approach, we were able to identify proteins that are linked to causal genes, enabling the potential development of functional molecular markers for crop improvement in nutrient use efficiency.     

Maternal control of seed weight in rapeseed (Brassica napus L.): the causal link between the size of pod (mother,source) and seed (offspring,sink)

Seed size/weight is one of the key traits related to plant domestication and crop improvement. In rapeseed (Brassica napus L.) germplasm, seed weight shows extensive variation, but its regulatory mechanism is poorly understood. To identify the key mechanism of seed weight regulation, a systematic comparative study was performed. Genetic, morphological and cytological evidence showed that seed weight was controlled by maternal genotype, through the regulation of seed size mainly via cell number. The physiological evidence indicated that differences in the pod length might result in differences in pod wall photosynthetic area, carbohydrates and the final seed weight. We also identified two pleiotropic major quantitative trait loci that acted indirectly on seed weight via their effects on pod length. RNA‐seq results showed that genes related to pod development and hormones were significantly differentially expressed in the pod wall; genes related to development, cell division, nutrient reservoir and ribosomal proteins were all up‐regulated in the seeds of the large‐seed pool. Finally, we proposed a potential seed weight regulatory mechanism that is specific to rapeseed and novel in plants. The results demonstrate a causal link between the size of the pod (mother, source) and the seed (offspring, sink) in rapeseed, which provides novel insight into the maternal control of seed weight and will open a new research field in plants.     

Oviposition by the African migratory locust, Locusta migratoria migratorioides, in a crop environment in South Africa

Oviposition by the African migratory locust, Locusta migratoria migratorioides (Orthoptera: Acrididae), was studied in maize and wheat crops on the Orange Free State Highveld. Maize was shown to be the most important oviposition habitat with peak laying taking place in autumn and early winter when highest pod densities were recorded. Laying was mainly concentrated along the middle of the crop interrows in maize and within clearings in the wheat crop. Despite the uniform layout of these crops, the distribution of egg pods was found to be aggregated. Non-reproductive behaviour, such as locust aggregation, basking and feeding, as well as environmental factors appeared to influence the distribution of egg pods in these crops. Secondary selection for optinum soil moisture and compaction on the laying site enhanced the aggregation of pods.     

Genetic basis of variation of yield, and yield components in mungbean (Vigna radiata (L.) Wilczek)

Additive, dominance, and epistasis genetic basis of seed yield per plant, number of pods per plant, number of seeds per pod, and 1000 seed weight in mungbean (Vigna radiata (L.) Wilczek) have been examined, using Triple Test Cross (TTC) analysis. The material for TTC test was evaluated in two seasons i.e., kharif (July-October) and spring/summer (March-June), at the research station of the Nuclear Institute for Agriculture and Biology (NIAB), Faisalabad, Pakistan. Epistasis was present significantly for number of pods per plant and number of seeds per pod when grown in the spring/summer season (March to June). Partition of epistasis showed that additive x additive ('i' type) interaction was an important component of number of pods per plant, and number of seeds per pod was found to be of both types 'i' type, and additive x dominance, and dominance x dominance ('j' and 'l' type) interactions. This indicated that epistasis might be a non-trivial factor in the inheritance of pods per plant, and seeds per pod in mungbean. The expression of epistasis was influenced differentially by particular genotypes, indicating that a limited number of genotypes may not be sufficient to detect non-allelic interactions for a trait in mungbean. Additive and dominance genetic components were significant for all four traits in kharif season (July to October) but only for seed yield and 1000 seed weight in spring/summer season. This suggests that the genes controlling seed yield per plant, and 1000 seed weight are equally sensitive to the environment. The predominance additive gene action in those traits is not significantly influenced by epistasis, suggesting that improvement of the traits can be achieved through standard selection procedures.     

Proteomic analysis of changes in mitochondrial protein expression during fruit senescence

Fruit senescence has been reported to be an oxidative phenomenon, but the detailed mechanisms by which ROS regulate this process remain largely unknown. Here we show that senescence process of apple fruit was concomitant with the dynamic alterations in the mitochondrial proteome. Mitochondrial proteins involved in tricarboxylic acid cycle, electron transport chain, carbon metabolism, and stress response were found to be differentially expressed during fruit senescence. Alleviating oxidative stress by lowering the ambient oxygen concentration noticeably decreased the number of changed proteins and delayed fruit senescence, indicating the involvement of ROS in this process. To further investigate the regulatory effect of ROS on senescence process, we analyzed the mitochondrial proteome variations upon exposure to high oxygen (100%), which induces oxidative stress and accelerates fruit senescence. High oxygen treatment led to a further identification of differentially expressed proteins such as mitochondrial manganese superoxide dismutase, an antioxidant scavenging superoxide radicals produced in the mitochondria. Activity of manganese superoxide dismutase was reduced after high oxygen exposure, accompanied by an increase in oxidative protein carbonylation (damaged proteins). These data suggest that ROS may regulate fruit senescence by changing expression profiles of specific mitochondrial proteins and impairing the biological function of these proteins.