Journal of Crop Production and Processing

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Motallebi-Azar A R, Shojaei H, Amani M. Optimization of Culture Medium for Propagation and Rooting of Greenhouse Cucumber at Various Nitrogen and Sucrose Levels. Journal of Crop Production and Processing 2025; 15 (4) :99-118
URL: http://jcpp.iut.ac.ir/article-1-3393-en.html

Extended Abstract
Introduction: Cucumber is a mineral-rich vegetable known for its detoxifying and antioxidant properties. While traditional cucumber varieties produce both male and female flowers, greenhouse cucumbers mainly produce female flowers. Seed propagation is common, but the high cost of hybrid seeds and the labor-intensive production process make in-vitro micropropagation a viable alternative. This study aims to optimize tissue culture techniques for cucumber to enhance micropropagation, improve plantlet regeneration, and address hybrid seed production issues while identifying disease-resistant and salt-tolerant varieties.

Materials and Methods: This study investigated the effects of different concentrations of total nitrogen (50, 55, 60 (control), 65, and 70 mM) and sucrose (20, 30, and 40 g l-1) in a factorial design within a completely randomized design with four replications on the propagation and rooting of the greenhouse cucumber cultivar Sultan, using lateral and terminal bud explants. The measured traits in this study included: (1) the percentage of established explants (the number of developed samples relative to the total number of explants) after 15 days; (2) the percentage of explants that developed nodes (the number of nodes producing lateral shoots relative to the total explants cultured) after 30 days; (3) the average shoot length after 30 days; (4) the number of shoots produced per explant after 30 days; (5) the number of leaves produced per shoot after 15 and 30 days; (6) the percentage of shoots producing roots (the ratio of rooted shoots to the total shoots cultured) after 15 and 30 days; (7) the root length per shoot (average length of roots produced on a shoot) after 30 days; and (8) the success rate of transfer (the number of developed plantlets relative to the number of plantlets transferred to mist conditions) after 15 days. The analysis of variance (ANOVA) of the data was conducted in a factorial experiment within a completely randomized design with three replications. The means of the treatments were compared using Duncan’s multiple range test at a 5% significance level. Data analysis was performed using SPSS software, and graphs were created using Excel software.

Results and Discussion: The highest fresh and dry weights of the shoot were observed in the medium containing 40 g l-1 of sucrose, while the highest fresh weight of the shoot and callus was recorded in 70 mM of nitrogen. The culture media containing 20 and 30 g l-1 of sucrose and 50 to 70 mM of nitrogen were identified as suitable for shoot length. The maximum number of nodes per shoot was obtained in the medium containing 30 g l-1 of sucrose, and the maximum shoot length was achieved at sucrose concentrations of 20 and 30 g l-1. At nitrogen concentrations of 50, 60, and 70 mM, the fresh weight of the shoot significantly increased. The lateral buds derived from terminal bud culture were rooted in MS medium without hormones, supplemented with five nitrogen concentrations, three sucrose concentrations, and 0.1 mM of IBA. The highest number of nodes per shoot, as well as the maximum shoot and root lengths, were produced in the culture medium containing 20 and 30 g l-1 of sucrose. The data clearly demonstrate the significant effects of sucrose and total nitrogen concentrations on the growth characteristics of cucumber in tissue culture. Sucrose plays a key role as a carbon source in tissue culture media, and its different concentrations have significant impacts on the number of nodes, shoot length, and diameter of the callus. Notably, an increase in sucrose concentration consistently 

Received: Oct. 23, 2025; Revised: Dec. 03, 2025; Accepted: Dec. 04, 2025; Published Online: Dec. 27, 2025.
* Corresponding Author: Minaamani@tabrizu.ac.ir    
led to a decrease in the number of nodes and shoot lengths, which may be due to its negative effects on the plant’s metabolic processes. Total nitrogen, as an essential element for growth, was identified in this study as a key factor in increasing the fresh weight of the shoots; however, its effect was less pronounced compared to sucrose. These results may be attributed to the sufficiency of total nitrogen at the tested concentrations. Transferring the plantlets to pots and gradually changing conditions from in-vitro to greenhouse environments are crucial for reducing stress and enhancing the viability of the plants. The use of sterilized perlite as a growing medium improves aeration and prevents fungal diseases, while maintenance methods in misting conditions effectively preserve moisture and increase the survival rate of the plantlets.

Conclusions: The results of this study clearly demonstrate the significant effects of sucrose and total nitrogen concentrations on the growth characteristics of cucumber in tissue culture. Sucrose plays a key role as a carbon source in tissue culture media, and its different concentrations have significant impacts on the number of nodes, shoot length, and diameter of the callus. Notably, an increase in sucrose concentration consistently led to a decrease in the number of nodes and shoot lengths, which may be due to its negative effects on the plant’s metabolic processes. Total nitrogen, as an essential element for growth, was identified in this study as a key factor in increasing the fresh weight of the shoots; however, its effect was less pronounced compared to sucrose. These results may be attributed to the sufficiency of total nitrogen at the tested concentrations. Transferring the plantlets to pots and gradually changing conditions from in-vitro to greenhouse environments are crucial for reducing stress and enhancing the viability of the plants. The use of sterilized perlite as a growing medium improves aeration and prevents fungal diseases, while maintenance methods in misting conditions effectively preserve moisture and increase the survival rate of the plantlets.