GmWRKY53, a water- and salt-inducible soybean gene for rapid dissection of regulatory elements in BY-2 cell culture

Abstract

Among abiotic stress conditions, drought is the main culprit that causes huge crop losses worldwide. Therefore, engineering drought tolerance in plants is of major economic importance. Many of the experimental systems that are used in attempts to improve drought tolerance, for example the generation of transgenic plants, are time consuming. For this reason, systems of reduced complexity that allows rapid screening of genes and promoters for potential use in projects aimed at improving drought tolerance are desired. One such system could be the tobacco (Nicotiana tabacum L.) Bright Yellow-2 (BY-2) cell line. 1,2BY-2 cells can be easily transformed to a very high efficiency using Agrobacterium tumefaciens and the cells subsequently can respond to stimuli such as the plant hormone methyl jasmonate (JA).3 The use of BY-2 cells also has potential advantages over other cell cultures, such as a highly synchronized growth rate, good transformation after protoplastation or particle bombardment, lack of leaky expression for promoter studies and easy monitoring and analysis. BY-2 cell system is also an excellent tool for sub-cellular localization studies. These characteristics make BY-2 cells an excellent system for many procedures.2 This system of reduced complexity therefore has the potential to be used to identify and better understand regulatory networks in response to water deficit/drought conditions.A number of reports have described using the BY-2 cell system for promoter analysis utilizing either β-glucuronidase 3,4 orDrought is the major cause of crop losses worldwide. Water stress-inducible promoters are important for understanding the mechanisms of water stress responses in crop plants. here we utilized tobacco (Nicotiana tabacum L.) Bright Yellow 2 (BY-2) cell system in presence of polyethylene glycol, salt and phytohormones. Extension of the system to 85 mM naCl led to inducibility of up to 10-fold with the water stress and salt responsive soybean GmWRKY53 promoter. upon aBa and Ja treatment fold inducibility was up to 5-fold and 14-fold, respectively. thus, we hypothesize that GmWRKY53 could be used as potential model candidate for dissecting drought regulatory elements as well as understanding crosstalk utilizing a rapid heterologous system of BY-2 culture.GmWRKY53, a water-and salt-inducible soybean gene for rapid dissection of regulatory elements in BY-2 cell culture

The interactome of soybean GmWRKY53 using yeast 2-hybrid library screening to saturation

“…GmWRKY53 is fully consistent with not only the identity of the interacting proteins (VQ) but also the large number of interactors from this family (19).…”

“…GmWRKY53 is up-regulated at the mRNA level as a response to water stress, 19 salt, 18 and Fusarium virguliforme infection. 20 This suggests that GmWRKY53 is part of several stressrelated signalling webs including biotic and abiotic stresses.…”

“…Using transgenic tobacco (Nicotiana tabacum L.) Bright Yellow 2 (BY-2) cells we have previously shown that the GmWRKY53 promoter directs salt and jasmonate inducible expression and to a lesser degree ABA inducible expression. 19 This suggests that the GmWRKY53 transcription factor is part of several stress-related signalling webs. Consistent with this, transcriptome analysis of the soybean root response to Fusarium virguliforme revealed that GmWRKY53 is the most strongly up-regulated WRKY transcription factor gene during both resistant and susceptible interactions.…”

Molecular identification and interaction assay of the gene (OsUbc13) encoding a ubiquitin-conjugating enzyme in rice

“…The non-fusion construct pMDC43 was used as a control. Here, the tobacco cell system as a model system was selected to study subcellular localization of OsUbc13 for its potential advantages over other cell cultures such as highly synchronized growth rate, high transformation efficiency, and easy monitoring and analysis (Geelen and Inzé, 2001;Tripathi et al, 2013). The data revealed that OsUbc13 was probably localized in the plasma The detection was performed three times and the data were presented as mean±standard deviation (SD).…”