Comparison of Different Methods for Separation of Haploid Embryo Induced through Irradiated Pollen and Their Economic Analysis in Melon ( Cucumis melo

Irradiated pollen technique is the most successful haploidization technique within Cucurbitaceae. After harvesting of fruits pollinated with irradiated pollen, classical method called as “inspecting the seeds one by one” is used to find haploid embryos in the seeds. In this study, different methods were used to extract the embryos more easily, quickly, economically, and effectively. “Inspecting the seeds one by one” was used as control treatment. Other four methods tested were “sowing seeds direct nutrient media,” “inspecting seeds in the light source,” “floating seeds on liquid media,” and “floating seeds on liquid media after surface sterilization.” Y2 and Y3 melon genotypes selected from the third backcross population of Yuva were used as plant material. Results of this study show that there is no statistically significant difference among methods “inspecting the seeds one by one,” “sowing seeds direct CP nutrient media,” and “inspecting seeds in the light source,” although the average number of embryos per fruit is slightly different. No embryo production was obtained from liquid culture because of infection. When considered together with labor costs and time required for embryo rescue, the best methods were “sowing seeds directly in the CP nutrient media“ and ”inspecting seeds in the light source.

Kavunda (cucumis melo var. inodorus) ışınlanmış polenle uyartılmış haploid embriyoların ayrılmasında kullanılabile

TEZ7374Tez (Yüksek Lisans) -- Çukurova Üniversitesi, Adana, 2009.Kaynakça (s.54-60) var.xii, 61 s. : rnk.res. ; 29 cm.This study was conducted between the years 2007-2009 in Horticulture Department, Faculty of Agriculture, University of Çukurova. Genotype Y2 and Y3 selected from backcross three (BC3) population of Yuva melon genetic resources collected on the Department of Horticulture are allocated. In this study, different methods for embryo rescue were tested to find an easier, fast, economical and effective method. ""Inspecting the seeds one by one"" was used as control treatment. Other four methods tested are ""sowing seeds direct nutrient media"", ""inspecting seeds on the light"", ""floating seeds on liquid media"" and "" floating seeds on liquid media after surface sterilization"". Overcome of this study show that, there is no significant difference statistically between the methods ""Inspecting the seeds one by one"", ""sowing seeds direct CP nutrient media"" and ""inspecting seeds on the light"" although the average number of embryos per fruit are slightly different (3.3, 2.4 and 3.1 respectively). No embryo production was obtained from liquid culture due to infection. When labor costs and time requied for embryo rescue taking into account, the best methods are ""sowing seeds direct CP nutrient media"" and ""inspecting seeds on the light"".Bu çalışma 2007-2009 yılları arasında Çukurova Üniversitesi Ziraat Fakültesi Bahçe Bitkileri Bölümünde yürütülmüştür. Materyal olarak Bahçe Bitkileri Bölümü bünyesinde toplanan kavun genetik kaynağındaki Yuva tipi geriye melez 3 (GM3) populasyonundan Y2 ve Y3 nolu genotipler kullanılmıştır. Bu çalışmada embriyoların daha kolay, hızlı, ekonomik ve etkili şekilde çıkartılabilmesi için farklı yöntemler denenmiştir. Çalışmada kontrol uygulaması olarak ""tohumların tek tek açılması"" yöntemi kullanılmış; bunun yanında, ""tohumların doğrudan besin ortamına ekimi"", ""tohumlara ışıkta bakma"" , ""tohumların sıvı besin ortamına ekimi"" ve ""tohumlara yüzey sterilizasyonu yapılarak ekimi"" yöntemi olmak üzere olmak üzere beş farklı yöntem denenmiştir. Deneme sonucunda, meyve başına elde edilen ortalama embriyo sayıları tanık olarak kullanılan tek tek açma yönteminde 3.3 ile en yüksek çıkmış, bunu sırayla 3.1 adet ile tohumlara ışıkta bakarak ve 2.4 adet ile tohumların hepsinin CP besin ortamına ekimi yöntemleri izlemiş fakat bu üç uygulama arasındaki farklılıklar istatistiksel olarak önemli çıkmamıştır. Tüm tohumların sıvı ortama ekimi uygulamasında ise hiç embriyo gelişimi sağlanamamış ve yüksek oranda enfeksiyon sorunu meydana gelmiştir. Gerekli süre ile birlikte işçilik maliyeti dikkate alındığında ise en iyi uygulamanın tohumların ışıkta bakılarak ayrılması yöntemi olmuş, bunu tohumların doğrudan CP besin ortamına ekimi izlemiştir.Bu çalışma Ç.Ü. Bilimsel Araştırma Projeleri Birimi Tarafından Desteklenmiştir. Proje No:ZF2008YL1

Effects of different genotypes and gamma ray doses on haploidization using irradiated pollen technique in squash

Fourteen genotypes and 3 different gamma ray doses were tested to develop an efficient haploidization protocol in squash. For this purpose, male flowers collected 1 day before anthesis were irradiated with 150, 200, and 300 Gy gamma ray doses, and female flowers were pollinated with the irradiated pollens the next day. In the first year of the study, 1858 embryos were obtained from 219 fruits. While 1358 of these were found in fruits irradiated with a 150 Gy gamma dose, the remaining 500 embryos were found in fruits irradiated with 200 Gy. From fruits irradiated with 150 and 200 Gy gamma doses, 9.12 and 3.53 haploid embryos per 100 seeds were obtained, respectively. While Genotype 3 was the most successful genotype with 12.42 embryos per 100 seeds, the minimum embryo numbers were obtained from Genotype 4 with 1.46 embryos per 100 seeds. In the second experimental year, 8 genotypes and the same gamma doses were used, and 2625 haploid and 1378 diploid embryos were obtained from 217 fruits. At irradiation doses of 150, 200, and 300 Gy, 2010, 539, and 76 haploid embryos were found, respectively. Genotype 6 was the most successful genotype with 13.35 embryos per 100 seeds.Fourteen genotypes and 3 different gamma ray doses were tested to develop an efficient haploidization protocol in squash. For this purpose, male flowers collected 1 day before anthesis were irradiated with 150, 200, and 300 Gy gamma ray doses, and female flowers were pollinated with the irradiated pollens the next day. In the first year of the study, 1858 embryos were obtained from 219 fruits. While 1358 of these were found in fruits irradiated with a 150 Gy gamma dose, the remaining 500 embryos were found in fruits irradiated with 200 Gy. From fruits irradiated with 150 and 200 Gy gamma doses, 9.12 and 3.53 haploid embryos per 100 seeds were obtained, respectively. While Genotype 3 was the most successful genotype with 12.42 embryos per 100 seeds, the minimum embryo numbers were obtained from Genotype 4 with 1.46 embryos per 100 seeds. In the second experimental year, 8 genotypes and the same gamma doses were used, and 2625 haploid and 1378 diploid embryos were obtained from 217 fruits. At irradiation doses of 150, 200, and 300 Gy, 2010, 539, and 76 haploid embryos were found, respectively. Genotype 6 was the most successful genotype with 13.35 embryos per 100 seeds

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

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