نوع مقاله : مقاله پژوهشی

نویسندگان

• مریم رحیمی ¹
• سیامک کلانتری ²
• اکبر دیزجی ³
• مجید شکرپور ⁴

¹ گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه تهران، کرج- ایران

² گروه علوم باغبانی، دانشکده کشاورزی ی، دانشگاه تهران ، کرج- ایران

³ گروه گیاهپزشکی، دانشکده کشاورزی، دانشکدگان کشاورزی و منابع طبیعی، دانشگاه تهران، کرج، ایران

⁴ گروه علوم باغبانی، دانشکده کشاورزی، دانشگاه تهران، کرج - ایران

چکیده

کلیدواژه‌ها

• الایزا
• زعفران
• ویروس زدایی
• RT-PCR
• RT-qPCR

عنوان مقاله [English]

Production of saffron plants free of saffron latent virus using meristem culture and thermotherapy

نویسندگان [English]

• Maryam Rahimy ¹
• Siamak Kalantari ²
• Akbar Dizadji ³
• Majid Shokrpour ⁴

کلیدواژه‌ها [English]

• ELISA
• RT-PCR
• RT-qPCR
• Virus elimination
• saffron latent virus

Extended Abstract

Introduction

      Saffron, often referred to as "Red Gold," is one of the most expensive medicinal plants globally, with a history of use spanning over 4,000 years. Iran is the leading producer of saffron, contributing more than 90% of the global supply. However, the emergence of Saffron latent virus (SaLV), a newly identified Potyvirus species, has severely impacted saffron cultivation in Iran, with infection rates exceeding 70%. SaLV compromises the quality and yield of saffron, necessitating effective virus elimination strategies. Common methods for producing virus-free plants include meristem culture, thermotherapy, chemotherapy, and their combinations. For instance, thermotherapy combined with meristem culture has successfully eliminated viruses such as bean yellow mosaic virus (BYMV) in gladiolus and apple chlorotic leaf spot virus (ACLSV) in apple cultivars. This study explores the efficacy of meristem culture, thermotherapy, and their combination in eliminating SaLV from saffron plants.

Material and Methods

      A total of 530 leaf tissue samples were collected from saffron plants cultivated at the Horticulture Research Center, University of Tehran. Samples were screened for SaLV using a Potyvirus genus-specific antibody (RT-0573/1, DSMZ, Germany). Positive samples (absorbance >3× healthy control at 405 nm) were subjected to meristem culture (0.3, 0.5, and 0.7 mm) and thermotherapy (50 °C for 60 min; 40 °C for 60 min; 28 °C for 21 days). Corm tissues from ELISA-positive plants were treated, while ELISA-negative corms were cultivated without treatment. Post-treatment, SaLV presence was assessed using ACP-ELISA, RT-PCR, and RT-qPCR. Data were analyzed using Tukey’s test in R 4.3.2.

Results

    Totally, 420 out of 530 samples (80.7%) tested positive for SaLV. Meristem culture and thermotherapy significantly influenced the percentage of virus-free plants (p < 0.05). Smaller meristem sizes yielded higher SaLV-free rates: 83.73% and 81.29% for 0.3 mm, compared to 38.5% and 30.91% for 0.7 mm, based on ELISA and RT-PCR, respectively. Thermotherapy at 50 °C for 60 min achieved the highest SaLV-free rate (71.36% and 66.36%), while 40 °C for 60 min yielded the lowest (50.98% and 46.47%). The combined treatment of 0.3 mm meristem culture and thermotherapy at 50 °C for 60 min resulted in 100% SaLV-free plants, as confirmed by ELISA, RT-PCR, and RT-qPCR.

Conclusion

    This study demonstrates that the combination of 0.3 mm meristem culture and thermotherapy at 50 °C for 60 min effectively eliminates SaLV from saffron plants.  This approach provides a robust method for producing virus-free saffron cultivars, supporting sustainable saffron production and propagation. Future research should explore the scalability and economic feasibility of this method for large-scale applications.