Assessment of possible haploid plant production in tomato (Lycopersicon esculentum Mill. cvs. Berlina, Petoperide, and Microtome) via anther and microspore culture

Ahmadi, Behzad and Asghari Zakaria, Rasool and Enayati Shariatpanahi, Mehran and Zare, Nasser and Azadi, Pejman (2015) Assessment of possible haploid plant production in tomato (Lycopersicon esculentum Mill. cvs. Berlina, Petoperide, and Microtome) via anther and microspore culture. Doctoral thesis, university of Mohaghegh Ardabili.

Text (بررسی امکان تولید گیاهان هاپلوئید گوجه فرنگی (Lycopersicon esculentum Mill. ارقام برلینا، پتوپراید و میکروتوم) به روش كشت بساك و ميكروسپور) Behzad Ahmadi.pdf Download (316kB)

Abstract

In this study, embryogenic response of isolated microspores and anthers were assessed in three cultivars of tomato (Berlina, Petoperide, and Microtom). Results of isolated microspore culture in the NLN medium supplemented with 13% sucrose indicated that microspore-derived structures with more than 10 nuclei were only observed in cv. "Berlina" and in the cultures were pretreated for 1 or 2 days at 4°C and then 2 days at 30°C. In addition, cold pretreatment (4°C) for 1 or 2 days and then 2 days treatment at 30°C efficiently induced formation of microspore-derived structure with 9-10 nuclei in both cultivars tested. Structures with more than 5 nuclei were not detected in the cultures were treated at 30°C for 10 days. In cv. "Berlina", structures with 9-10 nuclei were observed in the presence of 25 mg l-1 colchicine treatment for 48h whereas in cv. Petoperide, structures with more than 8 nuclei were not observed in all levels tested. Symmetrical nucleus divisions were not induced in cv. "Microtom" by all inductive stressors applied. Results of anther culture in cv. "Berlina" in solid medium indicated that callogenesis was significantly enhanced in the combination of 0.5 mg l-1 IAA and 0.5 mg l-1 Kin and in cv. "Petoperide", in the presence of 0.5 mg l-1 IAA and 1.0 mg l-1 Kin. Combination of 2, 4-D and BAP also affected callogenesis of cultured anthers so that the highest callusing in cv. "Berlina" was achieved in the cultures containing 2.0 mg l-1 2, 4-D and 2.0 mg l-1 BAP and in the presence of 2.0 mg l-1 2, 4-D and 1.0 mg l-1 BAP in cv. "Petoperide". Plantlet regeneration from calli was observed in cv. "Berlina" and in the cultures containing 50 mg l-1 chitosan. Culture of anthers in the two-layered induction medium also efficiently induced sporophytical divisions in microspores and ultimately led to the formation of globular embryos. Microspore-derived embryos were only detected in the B5 medium supplemented with 2% sucrose, cold treatment (4°C) for 1 and 2 days and heat treatment (30°C) for 2 and 5 days. Based on our knowledge, this study is the first report of embryo regeneration from isolated microspores of tomato. It can be concluded that, microspore embryogenesis could be induced in tomato when appropriate cultivar, optimum culture medium and duration of cold and heat treatment were selected. Molecular analysis indicated that SERK1 and SERK2 genes play important role in microspore embryogenesis induction and plant regeneration.

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