Use of Tissue Culture Techniques for Producing Virus-Free Plant in Garlic and Their Identification through Real-Time PCR

Abstract

This study was performed for comparison of meristem culture technique with shoot tip culture technique for obtaining virus-free plant, comparison of micropropagation success of two different nutrient media, and determination of effectiveness of real-time PCR assay for the detection of viruses. Two different garlic species (Allium sativum and Allium tuncelianum) and two different nutrient media were used in this experiment. Results showed that Medium 2 was more successful compared to Medium 1 for both A. tuncelianum and A. sativum (Kastamonu garlic clone). In vitro plants obtained via meristem and shoot tip cultures were tested for determination of onion yellow dwarf virus (OYDV) and leek yellow stripe virus (LYSV) through real-time PCR assay. In garlic plants propagated via meristem culture, we could not detect any virus. OYDV and LYSV viruses were detected in plants obtained via shoot tip culture. OYDV virus was observed in amount of 80% and 73% of tested plants for A. tuncelianum and A. sativum, respectively. LYSV virus was found in amount of 67% of tested plants of A. tuncelianum and in amount of 87% of tested plants of A. sativum in this study.

Comparitive Study on Volatile Aroma Compounds of Two Different Garlic Types (Kastamonu And Chinese) Using Gas Chromatography Mass Spectrometry (Hs-Gc/Ms) Technique

“…Therefore, pests and diseases can be carried via contaminated materials. Garlic is sexually sterile and therefore, is propagated vegetatively (Taşkın et al, 2013). In recent years, Chinese garlic clone was seen as an alternative to Kastamonu garlic clone both in Turkish markets and production areas.…”

“…Kastamonu and Chinese garlic samples were provided from Taşköprü-Kastamonu region of Turkey (Figure 1), and from Turkish market, respectively. Kastamonu garlic clone is suitable for winter consumption, can be stored for a long time, and suitable for processing due to its dry matter content (Türkeş, 1978;Taşkın et al, 2013). In a study carried out by Artık and Poyrazoğlu (1994), properties of Kastamonu garlic type were determined as: average bulb weight: 30.85 g (minimum: 12.56 g, maximum: 50.18 g), average clove number: 13 (minimum: 6, maximum: 20), average big clove number: 5 (minimum: 1, maximum: 13), average small clove number: 8 (minimum: 2, maximum: 16), average big clove weight: 2.697 g (minimum: 0.64 g, maximum: 6.7 g), average small clove weight: 1.643 g (minimum: 0.361 g, maximum: 3.759 g), average dry matter: 39.07%, average pH: 6.30, average total sugar: 40.15%.…”

“…Kastamonu garlic type is suitable for winter consumption, and thus can be stored for a long period of time. Hence it is suitable for processing because of its dry matter content (Türkeş, 1978;Taşkın et al, 2013). China ranks first among the world garlic-producer countries with 833 400 ha area and a total production yeild of 19 219 939 tones (Anonymous, 2011).…”

Eliminasi Onion yellow dwarf virus melalui Kultur Meristem Tip pada Bawang Merah (Allium ascalonicum L.)

“…Hal ini didasari oleh asumsi bahwa bagian kubah apikal bebas dari infeksi virus atau mengandung sedikit konsentrasi virus (Wang dan Hu, 1980). Metode ini telah diaplikasikan oleh Verbeek et al (1995) dan Taşkin et al (2013) pada bawang putih dan berhasil mendapatkan tanaman yang bebas dari infeksi OYDV. Penelitian Kasutjianingsih et al (2011) menunjukkan bahwa aplikasi in vitro Rizobakteri sebagai agen hayati dapat menurunkan keparahan penyakit layu Fusarium.…”

“…Pada umumnya, sitokinin digunakan dalam kultur meristem tip. Aplikasi sitokinin (2-ip, BA dan kinetin) pada konsentrasi tinggi dengan kisaran 0.5-2.0 mg L -1 dilaporkan dapat menginduksi terjadinya multiplikasi tunas pada meristem tip bawang putih, baik pemberian dalam bentuk kombinasi auksin-sitokinin ataupun sitokinin tunggal (Taşkin et al, 2013;Gull et al, 2014 Persiapan eksplan diawali dengan membuang kulit terluar dan mencuci bersih umbi bawang merah, selanjutnya umbi dicuci bersih menggunakan deterjen selama 7 menit sebanyak 2 kali, dan dibilas dengan air mengalir, kemudian umbi direndam dalam larutan 0.04% streptomisin sulfat dan 0.16% mankozeb selama semalam, dan dibilas dengan akuades steril sebanyak 2-3 kali. Umbi disterilisasi dengan 1.05% NaOCl selama 20 menit.…”