Abstract

Maize Agrobacterium-mediated transformation efficiency has been greatly improved in recent years. Antioxidants, such as, cysteine, can significantly improve maize transformation frequency through improving the Agrobacterium infection efficiency. However, the mechanism underlying the transformation improvement after cysteine exposure has not been elucidated. In this study, we showed that the addition of cysteine to the co-cultivation medium significantly increased the Agrobacterium infection efficiency of hybrid HiII and inbred line Z31 maize embryos. Reactive oxygen species contents were higher in embryos treated with cysteine than that without cysteine. We further investigated the mechanism behind cysteine-related infection efficiency increase using transcriptome analysis. The results showed that the cysteine treatment up-regulated 939 genes and down-regulated 549 genes in both Z31 and HiII. Additionally, more differentially expressed genes were found in HiII embryos than those in Z31 embryos, suggesting that HiII was more sensitive to the cysteine treatment than Z31. GO analysis showed that the up-regulated genes were mainly involved in the oxidation reduction process. The up-regulation of these genes could help maize embryos to cope with the oxidative stress stimulated by Agrobacterium infection. The down-regulated genes were mainly involved in the cell wall and membrane metabolism, such as, aquaporin and expansin genes. Decreased expression of these cell wall integrity genes could loosen the cell wall, thereby improving the entry of Agrobacterium into plant cells. This study offers insight into the role of cysteine in improving Agrobacterium-mediated transformation of maize immature embryos.

Highlights

  • An efficient crop transformation method is one of the prerequisites for creating transgenic crops

  • We showed that cysteine could improve Agrobacterium infection efficiency in the maize inbred line Z31 and hybrid line HiII

  • The results showed that the up-regulated genes were mainly involved in the oxidation reduction process, and the down-regulated genes were mainly involved in the cell wall and membrane metabolism

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Summary

Introduction

An efficient crop transformation method is one of the prerequisites for creating transgenic crops. Widely used transformation methods include the particle bombardment method and the Agrobacterium-mediated method. Agrobacterium-mediated transformation has the advantages of simple operation, low cost, low foreign gene copy number, and the stable inheritance of foreign genes. The T-complexes formed between VirD2 and T-DNA enter the plant cell, where the T-complexes can bind the other Vir proteins (VirE2, VirE3, VirD5, and VirF) to form a super complex. This super complex can enter the plant cell nucleus and release the T-DNA for integration into the plant genome (Gelvin, 2010; Pitzschke and Hirt, 2010)

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