Abstract
Immature embryos of inbred maize (Zea mays) lines (H8183, H8184, and H8185) were used for Agrobacterium infection. We used the β-glucuronidase gene (GUS) as the target gene and the glufosinate resistance gene (bar) as the selection marker. We conducted research on several aspects, such as different genotypes, coculture conditions, screening agent concentrations, and concentrations of indole-3-butytric acid (IBA), 6-benzylaminopurine (6-BA), and ascorbic acid (Vc) in the differentiation medium. We optimized the genetic transformation system, and the obtained results indicated that among the three lines studied, the induction rate of H8185 was the highest at 93.2%, followed by H8184, with H8183 having the lowest induction rate (80.1%). The best coculture method was that using the N6 coculture medium layered with a sterile filter paper. Using orthogonal analysis, we found that the optimal combination of the three factors in the differentiation medium was A3 (1 mg mL−1 IBA), B3C1 (1.6 mg mL−1 6-BA), and D3 (1.5 mg mL−1 Vc). Through GUS staining analysis, Bar test-strip analysis, and polymerase chain reaction, five transgenic plants were finally obtained. This study established the optimal conditions for genetic transformation in maize.
Highlights
Maize, Zea mays L. (Gramineae family), originated in South America and is a globally important food crop
Agrobacterium-mediated genetic transformation was performed in immature embryos of three maize inbred lines, H8183, H8184, and H8185, and the results revealed that H8185 had the highest induction efficiency
At the embryogenic callus induction stage, induction rate and regeneration ability is controlled by genetic factors
Summary
Zea mays L. (Gramineae family), originated in South America and is a globally important food crop. (Gramineae family), originated in South America and is a globally important food crop. With the recent increasing demand, genetic modification via plant genetic engineering has become of great significance toward yield and quality improvement. There are various transformation methods, the efficiency of maize transformation is typically low owing to the lack of stable and efficient tissue culture and plant regeneration systems. Agrobacterium-mediated transformation has been widely used in dicotyledonous plants since 1983, when Zambryski successfully obtained transgenic tobacco. The application range of this method is limited for monocots, which are unnatural hosts of Agrobacterium. Studying the conditions affecting maize tissue culture and Agrobacterium-mediated genetic transformation is essential for cultivating high-yield, high-quality, and stress-resistant novel maize varieties
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