Phytomicrobiome Interactions and Sustainable Agriculture. Группа авторов

Phytomicrobiome Interactions and Sustainable Agriculture - Группа авторов


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their utility as important agricultural tools, there are inherent disadvantages to many of them and the search for novel, rapid and reliable techniques is an ongoing process within the scientific and agricultural communities.

      2.4.1 Gel‐Based Protein Separation Techniques

      The most frequently used technique with gel‐based technology is the 2DE as this method involves utilizing several characteristic features of proteins for separation such as proteins, isoelectric point, molecular mass, as well as solubility (Joshi 2017).

      Here in the foremost step, the proteins are subjected to a pH gradation throughout their gel run. Hence proteins separate and precipitate when their isoelectric constant is achieved. At this point at a particular pH, the overall charge of the protein neutralizes thereby precipitating the proteins. This is also known as isoelectric focusing (IEF). IEF is followed by 2DE separation where the proteins are segregated based upon their molecular weight done by the SDS‐PAGE technique (Gorg et al. 2004). The protein separation profile is then visualized using certain staining methods that stain the proteins against the gel. Then with the comparison with a standard set of proteins, one can verify and characterize the isolated proteins. The major setback of this technology is limited reproducibility and lower sensitivity, hence only a well abundant and most characterized protein separation is recommended (Bunai and Yamane 2005). 2D fluorescence differential gel electrophoresis (2DDIGE) is an alternative method where the separation of proteins is done on the same gel using differential fluorescent dyes and a comparative is set based upon their spot intensity (Dunn 1993).

      2.4.2 Non‐Gel Protein Separation Techniques

      A non‐gel separation technique is usotope coded affinity tag (ICAT) is in principle used to compare the two protein‐separated samples. It consists of a tag having one biotin group containing an isotope‐coded linker, which forms the heavy and light sections of a tag while the other group is a thiol. This technique is conveniently utilized in the integral membrane protein identification (Gygi et al. 1999). Isobaric tagging of relative and absolute quantification of protein (iTRAQ) uses the N terminus of the peptide chain along with the side chain which is tagged by the isobaric‐labeled tags. Within a given sample, this technique permits quantification (relative and absolute) of peptides from varied sources all at once (Ross et al. 2004).

Flowchart depicts proteomic approaches for phytomicrobiome analysis.

      Thus, microbes can be considered for positive exploitation of the plant growth promotion to enhance the growth of the plant. Microbes also make plants sustainable toward the abiotic as well as biotic stresses looking into progressive climate change. There have been many such types of research, where the positive associations have been well characterized using proteomic analysis and have further been utilized for the enhancement of a natural process.

      Enhancement of drought and salt resistance observed for the enhanced salt and drought resistance owing to the effects of plant growth–promoting bacteria as studied in L. perenne, also known as ryegrass. Unfortunately for this perennially relevant grass as turf and forage, it is highly sensitive to drought‐induced stress and high salinity. However, B. amyloliquefaciens, combined with hydrogels to prevent the soil from eroding, can help the grass sustain the drought‐induced stress by the plant (Su et al. 2017).

      In another example, a species of plant growth‐promoting bacteria that have been found to be colonizing the roots, stems, and leaf parts of the sugarcane is the species Gluconacetobacter diazotrophicus. This is one such example where the bacteria serve as a legume endophyte. However, the proteome interaction studies of this bacterium with the sugar cane are still not very deep. A study was performed to investigate the molecular aspects using MS proteomic analysis using N15 metabolic labeling of bacterial, root samples, and co‐cultures. Over 400 proteins were assessed out of which near about 78 combinations of involved proteins appeared relevant among the interaction model. A comparative analysis of the proteomic data thus derived revealed the proteins involved in fundamental roles of protein recognition. Additionally, 30 bacterial active proteins have been identified where 9 were categorically induced by plant signaling pathways. This is the first line of study for the G. diazotrophicus and sugar cane (Schirawski and Perlin 2018).

      The microbial action on plants can work both ways. Certain interactions work for the enhancement of the plant, however, some have a negative impact owing to such associations. Any plant maybe rendered vulnerable to disease owing to an undesirable microbial association or any abiotic stress situation which throws the plants defenses out of order. In the case of the pathogen associate


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