Genetic Transformation in Cucumber as Influenced by Inoculation Time and CoCultivation Period
DOI:
https://doi.org/10.69996/ijari.2024020Keywords:
genetic transformation, cucumber, abiotic stressAbstract
The LBA4404 strain of Agrobacterium tumefaciens was utilized to assess the impact of inoculation time and co-cultivation period on the genetic transformation of cucumber (var. Shital) for abiotic stress tolerance. This strain carries the CIPK sense gene. The transformation was performed on the leaf, nodal, and internodal calli. Histochemical analysis of GUS reporter gene expression in viable calli samples allowed for evaluation of transformation capacity. In the callus tissue, there was a noticeable patch that was positive for GUS (blue in color). In this inquiry, three things were considered. A total of three different kinds of explants—leaf, nodal, and internodal callus—made up factor A. Factor B included inoculation times of three and five minutes, respectively. Factor C included co-cultivation times of twenty-four and forty-eight hours. The number of calluses that tested positive for GUS (3.17% and 52.87%, respectively) increased with increasing immersion time in the bacterial suspension (from 3 to 5 minutes), while the number of calluses that tested negative for GUS decreased (2.15 and 34.88%, respectively) with decreasing immersion time.More time spent co-cultivating (48 hours) resulted in superior performance compared to a shorter period. After two days of co-cultivation (48 hours), a greater number of calluses (12.45%) and a higher proportion of calluses (51.87%), respectively, survived. After immersing leaf explants in an Agrobacterium suspension for 5 minutes and then transferring them to co-cultivation media for 48 hours, the maximum number of GUS +ve callus (4.04 and 67.38%, respectively) were achieved.
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