Comparative physiological, antioxidant and proteomic investigation reveal robust response to cold stress in Digitalis purpurea L.

BACKGROUND OF THE STUDY: Digitalis purpurea (L) is an important medicinal plant growing at Alpine region of Himalayas and withstands low temperatures and harsh climatic conditions existing at high altitude. It serves as an ideal plant system to decipher the tolerance to cold stress (CS) in plants from high altitudes. METHODS AND RESULTS: To understand the complexity of plant response to CS, we performed a comparative physiological and biochemical study complemented with proteomics in one-month-old D. purpurea grown at 25 °C (control) and 4 °C (CS). We observed an enhanced accumulation of different osmo-protectants (glycine betaine, soluble sugar and proline) and higher transcription (mRNA levels) of various antioxidant enzymes with an increased antioxidant enzyme activity in D. purpurea when exposed to CS. Furthermore, higher concentrations of non-enzymatic antioxidants (flavonoids, phenolics) was also associated with the response to CS. Differential proteomic analysis revealed the role of various proteins primarily involved in redox reactions, protein stabilization, quinone and sterol metabolism involved in CS response in D. purpurea.. CONCLUSION: Our results provide a framework for better understanding the physiological and molecular mechanism of CS response in D. purpurea at high altitudes.

Low temperature elicits differential biochemical and antioxidant responses in maize (Zea mays) genotypes with different susceptibility to low temperature stress.

Maize, a C4 sub-tropical crop, possesses higher temperature optima as compared to the C3 plants. Low temperature (LT) stress confines the growth and productivity of maize. In this context, two maize genotypes, LT tolerant Gurez local and LT susceptible Gujarat-Maize-6 (G-M-6) were analysed in present study for various osmolytes and gene expression of antioxidant enzymes including Ascorbate-glutathione (AsA-GSH) besides trehalose biosynthetic pathways. With the progressive LT treatment, Gurez local showed lesser accumulation of stress markers like hydrogen peroxide (H2O2) and malondialdehyde, a significant increase in osmoprotectants like free proline, total protein, total soluble sugars, trehalose, total phenolics and glycine betaine, and a significant reduction in the plant pigments as compared to the G-M-6. Additionally, Gurez local was found to possess a well-established antioxidant defense system as revealed from the elevated transcripts and enzyme activities of various enzymes of AsA-GSH pathway. Higher gene expression and enzyme activities were exhibited by superoxide dismutase, catalase and peroxidase besides the gene expression of trehalose biosynthetic pathway enzymes. Moreover, through principal component analyses, a positive correlation of all analysed parameters with the LT tolerance was noticed in Gurez local alone demarcating the genotypes on the basis of their extent of LT tolerance. Overall, the present study forms the basis for unravelling of LT tolerance mechanisms and improvement in the performance of the temperate maize. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s12298-021-01020-3.

Comparative proteomics and global genome-wide expression data implicate role of ARMC8 in lung cancer.

BACKGROUND: Cancer loci comprise heterogeneous cell populations with diverse cellular secretions. Therefore, disseminating cancer-specific or cancer-associated protein antigens from tissue lysates could only be marginally correct, if otherwise not validated against precise standards. MATERIALS AND METHODS: In this study, 2DE proteomic profiles were examined from lysates of 13 lung-adenocarcinoma tissue samples and matched against the A549 cell line proteome. A549 matched-cancer-specific hits were analyzed and characterized by MALDI-TOF/MS. RESULTS: Comparative analysis identified a total of 13 protein spots with differential expression. These proteins were found to be involved in critical cellular functions regulating pyrimidine metabolism, pentose phosphate pathway and integrin signaling. Gene ontology based analysis classified majority of protein hits responsible for metabolic processes. Among these, only a single non-predictive protein spot was found to be a cancer cell specific hit, identified as Armadillo repeat-containing protein 8 (ARMC8). Pathway reconstruction studies showed that ARMC8 lies at the centre of cancer metabolic pathways. CONCLUSIONS: The findings in this report are suggestive of a regulatory role of ARMC8 in control of proliferation and differentiation in lung adenocarcinomas.

Affinity proteomics led identification of vimentin as a potential biomarker in colon cancers: insights from serological screening and computational modelling.

Proteomic analysis using multiplex affinity reagents is perhaps the most reliable strategy to capture differentially expressed proteins that are slightly or immensely modified. In addition to expressional variation, it is comprehensively evident that the immunogenicity of a protein can be a deciding factor for instigating an inflammation afflicted-carcinogenesis. Considering both these factors, a simple and systematic strategy was designed to capture the immunogenic cancer biomarkers from sera of colorectal cancer patients. The affinity reagent, in the form of an antibody repertoire against the secretome of the HT29 cell line was used to grade the sera samples on the basis of the degree of immuno-reactivity and to capture differentially expressed antigens from the patient sera. Following affinity based 2DE-MALDI-TOF; the proteins were identified as (1) soluble vimentin; and (2) TGF-beta-inhibited membrane-associated protein (PP16B), in colon cancer sera and (3) keratin, type II cytoskeletal protein in rectal cancer sera. Pathway reconstruction and protein-protein networking of identified proteins predicted only Vimentin to be physically and genetically engaged in close proximity with the most established colorectal cancer associated tumorigenic pathways. Furthermore, our findings suggest that a possible surface stoichiometric shift in the structure of protein could be due to mutations in the coding sequence of Vimentin that may elicit its enhanced secretion possibly due to protein-hyperphosphorylation. Of the three proteins identified, only Vimentin showed higher expression in sera of colon cancer patients alone. Thus, it could be argued that vimentin might help in predicting individuals at higher risk of developing colon cancers. Our data are therefore suggestive of using vimentin as an antigen for tumor vaccination in an autologous set-up for colon cancers.

Alterations in stem sugar content and metabolism in sorghum genotypes subjected to drought stress.

Changes in stem sugar concentrations due to drought stress at the early reproductive stage were studied in seven sorghum (Sorghum bicolor (L.) Moench) genotypes that differ in their stem sugar storage ability. Total sap sugar concentration increased in most genotypes. ANOVA showed a significant contribution of genotype and treatment to the variation in sugar levels. Two genotypes showed little variation in total sugar levels at the fifth internode from the peduncle and five genotypes showed significant increases in total sugar levels under drought; these groups were used to compare sugar metabolism. Drought led to a decrease in catabolic sucrose synthase activity in both groups. Invertase activities increased significantly in two genotypes and correlated with the increase in reducing sugar concentrations under drought. Stem sugar hydrolysis probably had a role in osmotic adjustment under drought and correlated with retention of sap volume. However, the activities of sugar-metabolising enzymes did not correlate with their gene expression levels. After resuming irrigation, grain yields, stalk yields and juice volume at physiological maturity were lower in plants recovering from drought stress compared with the controls. In some genotypes, there were similar losses in grain yields and stem sugars due to drought, indicating photoassimilate source limitation; in other genotypes, grain yield losses were less than stem sugar losses in drought-exposed plants compared with the controls, suggesting mobilisation of sugars from the storage internodes to the developing panicle. Accumulation of stem sugars appears to be an adaptive strategy against drought stress in some sorghum genotypes.

Stem sugar accumulation in sweet sorghum - activity and expression of sucrose metabolizing enzymes and sucrose transporters.

Sugar metabolism was studied in sweet sorghum (SSV74) that is known to store sugars in the mature internodes and which is reported to give grain yields twice that of a grain sorghum variety (SPV1616). Comparison of sugar accumulation in these two varieties was carried out at three stages of growth and in the upper and lower internodes. In spite of large differences in the level of sugar accumulation, osmolarity of the sap did not vary as significantly in the two varieties. Significant contribution of variety, stage and internode position was seen for the variation observed in sugar content. Though the activities of sugar metabolizing enzymes namely sucrose synthase (in the synthesis and cleavage directions), sucrose phosphate synthase and invertase (cytoplasmic and vacuolar) also varied in a stage- and internode-specific manner in the two varieties, these enzymes did not contribute significantly to the variation observed in sugar content. Transcriptional expression of one sucrose synthase (SUC1), two sucrose phosphate synthase (SPS2 and SPS3) and a vacuolar invertase (INV3) gene were lower in sweet sorghum as compared to grain sorghum. Sweet sorghum also showed lower expression of two sucrose transporters (SUT1 and SUT4), which correlated to higher sugar accumulation in this variety. Differential expression of the sugar metabolizing enzymes and sucrose transporters in sweet and grain sorghum suggest a role for signaling molecules and transcription factors in regulating sugar accumulation observed in the mature internodes of sweet sorghum, which needs to be investigated.