Optimizing Factors Affecting Agrobacterium Tumefaciens Mediated Transformation Of Dwarf Rootstock Of Apple (Malus Domestica Borkh Cv Gami Almasi)

Mohseniazar, Mehdi; Nazeri, Sonbol; Malboobi, Mohammad Ali; Ghadimzadeh, Morteza; Barzgari, Abolfazl; Farokhzad, Ali Reza

Optimizing Factors Affecting Agrobacterium Tumefaciens Mediated Transformation Of Dwarf Rootstock Of Apple (Malus Domestica Borkh Cv Gami Almasi)

Document Type : Research Paper

Authors

Mehdi Mohseniazar ¹

Sonbol Nazeri ²

Mohammad Ali Malboobi ³

Morteza Ghadimzadeh ⁴

Abolfazl Barzgari ⁵

Ali Reza Farokhzad ⁶

¹ MSc of Biotechnology , Horticulture Department of Agriculture faculty, Urmia University

² Assistant professor of Biotechnology Department of agriculture faculty, Bu-Ali Sina University

³ Associated professor of National Institute of Genetic engineering and Biotechnology, Tehran, Iran

⁴ Associate professor of Plant breeding and Agronomy Department of agriculture faculty, Urmia University

⁵ Agricultural Biotechnology Research Institute of Tabriz – Iran

⁶ Assistant professor of Horticulture Department of Agriculture faculty, Urmia University

Abstract

Gami Almasi is a dwarf rootstock of apple (Malus domestica Bork) that originated from North West of Iran and has some good horticultural characteristics. Agrobacterium mediated gene transformation is a method that perform for introducing of gene into plant species and need for developed to new varieties. Present study performed for first time in this cultivar. Optimizing of factors that affected on transformation is necessary in transformation programs. In this research some factors that affected on transformation such as, Agrobacterium strain, bacteria suspension ‍‍concentration, infection time, Co-cultivation period and Agrobacterium infection medium type, were evaluated byinsertion of b-glucoronidase gene. In transformation experiments, infection by bacterial suspension in 0.5 OD at 600 wavelengths for 5 minutes, showed better results than other treatments. All regenerated shoots on selective medium tested by histochemical GUS assay and PCR. Agrobacterium tumefaciens strain GV3850, with 2 days co-cultivation period were better than GV3101 strain with 3 days, co-cultivation for transformation performance. LB medium had better effect than AB medium as Agrobacterium infection medium. Under the most suitable condition, 2% transformation ratio was observed.

Keywords

Agrobacterium

Apple

Gami

GusA

Strain

transformation