Production of Malus domestica L. virus free via meristem culture and the effects of pectin and phloroglucinol on its proliferation

Document Type : Research Paper

Authors

1 Professor, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

2 Assistant Professor, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

3 MSc graduate, Department of Plant Sciences, Faculty of Biological Sciences, Kharazmi University, Tehran, Iran.

10.22103/jab.2026.25499.1725

Abstract

Objective
Meristem culture is a method used to produce virus-free plants. This study aimed to eliminate common viruses affecting Malus domestica L. Additionally, the investigation focused on the effects of two compounds, pectin and phloroglucinol, on improving branching and rooting.  
Materials and methods
Lateral and apical bud explants were collected at three different time points in July, August, and September. The establishment of meristems was assessed using various concentrations of BA hormones. Explants derived from meristem growth were then transferred to an MS propagation medium supplemented with BA and GA3 hormones. After identifying the most suitable culture medium for propagation, the explants were moved to an optimal medium containing pectin and phloroglucinol to examine their effects on shoot length, shoot number, and leaf development. Rooting of elongated stems from the propagation stage was performed in MS½ medium with NAA and IBA hormones. Additionally, an experiment was conducted to evaluate the influence of phloroglucinol on rooting percentage. To confirm their virus-free status, regenerated plants from meristems were tested using RT-PCR.
Results
The results indicated that explants collected in the latter half of July in MS medium with a concentration of 1 mg L-1 BA showed the highest establishment percentage (26.66%). The optimal medium for promoting stem elongation contained 1 BA and 1 mg L-1 GA3. The highest leaf development was observed in MS medium with 1 mg L-1 BA and without GA3. The interaction between BA and GA₃ significantly affected the number of secondary branches, whereas no significant effect was observed on the number of primary branches. Adding pectin in the proliferation stage increased the stem length (45%) and the branch number (82.6%). Incorporation of phloroglucinol into the medium led to a 24% increase in stem length but a 39.13% decrease in branch quantity. The highest rooting percentage (34%) was observed in½ MS medium treated with 1 IBA plus 0.5 mg L-1 NAA. Phloroglucinol treatment diminished the rooting percentage while increasing the average number of roots per explant. RT-PCR analysis of the plant samples derived from meristem culture did not reveal any bands indicative of viral infection.
Conclusions
This research demonstrated that meristem culture is an effective technique for generating virus-free red-fleshed apple plants. Additionally, the RT-PCR method proved to be sufficiently precise for virus detection. Taking all factors into account, the application of pectin and phloroglucinol can serve as two dependable compounds to enhance branching during meristem culture.

Keywords

References

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Agricultural Biotechnology Journal
Volume 18, Issue 3
June 2026
Pages 27-54

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