Abstract

Drought stress poses a significant challenge to agriculture in the context of climate change, as it restricts chickpea yield and leads to financial losses. Chickpea (Cicer arietinum L.) is the second most widely farmed legume worldwide. This work aimed to examine the impact of altering the Sal1 gene, which encodes the 3’-phosphoadenosine 5’-phosphate phosphatase, on drought tolerance in chickpea. The CRISPR-based gene editing technique was employed for this purpose. Considering the constraints imposed by gene delivery systems and the laborious and expensive tissue culture procedure, we opted for the utilization of Agrobacterium rhizogenes. The designed sgRNA was inserted into the expression vector, pDW3873, which contains the nuclease null variant, nCas9 D10A, coupled to the PmCDA1 cytosine deaminase at the C-terminal, then injected into the A. rhizogenes strain K599. In this study, CRISPR/Cas9 was used to make precise changes to a specific genomic area in CaSal1 gene. Results demonstrated that a total of 6 chickpea plants were identified as transgenic plants. Their drought tolerance was enhanced and root showed double branches of the control but shorter in length. This article demonstrates that the CDB system effectively facilitates gene editing in a sterile-free environment, eliminating the requirement for tissue culture. Consequently, it is well-suited for the transformation of a substantial quantity of plants. This technique can also be utilized on several species that have the ability to produce root suckers.