OsRuvBL1a DNA Helicase Boost Salinity and Drought Tolerance in Transgenic Indica Rice Raised by In-Planta Transformation

Abstract

RuvBL, is a member of SF6 superfamily of helicases and is conserved among the various model systems. Recently rice homolog of RuvBL has been biochemically characterized for its ATPase and DNA helicase activities, however its involvement in stress is not been studied yet. This study reports the detailed functional characterization of RuvBL homolog of Oryza sativa, under abiotic stress through transgenic approach. An improved Agrobacterium-mediated in-planta transformation method was developed in indica rice to generate the transgenic lines and study was focused on optimization of factors to achieve maximum transformation efficiency. Overexpressing OsRuvBL1a transgenics showed enhanced tolerance under in vivo salinity stress as compared to WT plants. The physiological and biochemical analysis of the OsRuvBL1a transgenic lines showed better performance under salinity and drought stresses. Several stress responsive interacting partners of OsRuvBL1a were identified using Y2H method. Working mechanism for boosting the stress tolerance by OsRuvBL1a has been proposed in this study. This integration of OsRuvBL1a gene in rice genome using in-planta transformation method helped us to achieve the abiotic stress tolerant smart crop. This study is the first direct evidence to show the novel function of RuvBL in boosting abiotic stress tolerance in plants.

A Genotype-Independent, Simple, Effective and Efficient in Planta Agrobacterium-Mediated Genetic Transformation Protocol

“…A transformation efficiency of 35–41% was observed by [ 31 ] when in planta transformation of rice seed was carried out. However, the reported protocol involves pre-culturing of rice seeds for 24 h on half-strength MS medium and germination of the Agro-infected seeds on selection medium, leading to longer duration of the whole procedure.…”

Identification and characterization of RuvBL DNA helicase genes for tolerance against abiotic stresses in bread wheat (Triticum aestivum L.) and related species

“…13). In another earlier study also, expression of OsRuvBL1a differed under abiotic stresses (cold, heat and salinity; Sai et al 2017), which was later con rmed using transgenic approach (Sai et al 2021). Similar results were also reported in Arabidopsis (Holt et al 2002) and pigeon pea (Singh et al 2020).…”

“…RuvBL1 and RuvBL2 have been discovered and characterized only in few plant systems like Arabidopsis with one RuvBL gene-RUVBL1 (TIP49a/b) (Fritsch et al 2004; Holt et al 2002) and rice with four RuvBL genes-OsRuvBL1a, OsRuvBL1b, OsRuvBL2a, and OsRuvBL2b (Wang et al 2011;Sai et al 2017;Sai et al 2018;Sai et al 2021). RuvBL genes have also been utilized for the production of transgenic pigeonpea, to demonstrate the function of these genes in providing tolerance against abiotic stresses (Singh et al 2020).…”