Selection of optimum nutrient media for clonal micropropagation of the physalis alkekengi plant and creation of a virus-free plant

Abstract Background Physalis alkekengi is a medicinal plant rich in vitamins, flavonoids, and various bioactive substances. Its productivity is compromised by natural infection with the tobacco mosaic virus (TMV), which disrupts physiological processes. This study aimed to establish virus-free plantations of Physalis alkekengi using microcloning techniques. Methods Plant explants were cultured on a multicomponent Murashige-Skoog nutrient medium, supplemented with biologically active substances and growth regulators. Specific cultivation conditions, including optimal temperature, pH, humidity, and lighting, were maintained. The effects of nystatin and different concentrations of auxins (6 µM NAA-1-naphthylacetic acid and IBA-indole-3-butyric acid) on plant growth were evaluated. A PCR method with custom primers was developed to detect Ph-TMV at the molecular level. Virus-free plant cuttings were grown in soil under optimized conditions. Results The addition of nystatin to the nutrient medium enhanced the growth and development of Physalis alkekengi explants, though higher concentrations of nystatin had a suppressive effect. Optimal root growth and development were observed at an auxin concentration of 6 µM (NAA and IBA). The PCR method effectively detected the presence of TMV, facilitating the identification and elimination of infected plants. Under optimized conditions, virus-free Physalis alkekengi plants were successfully cultivated in soil. Discussion This study demonstrated that microcloning, along with precise control of nutrient medium components and environmental conditions, can effectively produce virus-free Physalis alkekengi plants. The beneficial effects of nystatin and auxins in specific concentrations were crucial for optimal plant growth. The PCR detection method proved reliable for identifying TMV-infected plants at the molecular level. Conclusion Microcloning, in combination with optimized nutrient media and growth conditions, is an effective strategy for cultivating virus-free Physalis alkekengi plants. This approach has the potential to enhance the productivity and medicinal value of this important plant species.