A simple and efficient seed-based approach to induce callus production from B73 maize genotype

Simeon O Kotchoni,Daniel P Russo,Lamine Baba-Moussa,Pacôme A Noumavo,Emma W Gachomo,Adolphe Adjanohoun,John Dell’Angelo

doi:10.4236/ajmb.2012.24039

Abstract

The wild type maize genotype, B73, is not amenable for callus production and an efficient protocol for B73 maize callus induction has never been reported up-to-date. Scientific efforts in producing B73 maize callus using all known callus inducible media have been unsatisfactory. Here we developed and described an efficient protocol for callus induction from B73 maize seedlings. The protocol is based on well known callus inducible media CM4C where we have sequentially subtracted some chemical compounds and added some new compounds mediating cell proliferations. This newly described protocol was able to induce callus production in a wide range of crop species including rice and soybean. We found that cell proliferation factors, NAA (auxin analog) and 2,4 D (auxin influx carrier) were not only very crucial but required for positive B73 maize callus induction. The absence of one or the other will lead to the failure of B73 maize callus production. The well known CM4C callus induction composition lacks NAA. Our findings will advance genetic studies of maize mutants generated from B73 genotype background.

Highlights

Callus production and transformation has been established as a powerful tool to study plant functional genomics, growth regulation, and plant cell morphogenesis
We found that cell proliferation factors, NAA and 2,4-D were very crucial but required for positive B73 maize callus induction
Different maize mutants that are highly susceptible to pathogens (Figures 3(B)-(D)), as well as two mutants that are resistant to pathogens

Summary

Introduction

Callus production and transformation has been established as a powerful tool to study plant functional genomics, growth regulation, and plant cell morphogenesis. Advances such as callus regeneration from explants and the production of diverse germplasms have provided useful tools for successful crop breeding programs. For still unknown reasons, production of callus from maize B73 genotype have proven difficult using various known callus induction media and production of B73 callus remains unsatisfactory. This bottleneck has resulted in often utilizing the maize inbred Hi II for the production of maize callus as an alternative to the B73 genotype. One experimental drawback is an inability to adequately evaluate mutant lines engineered in B73 background

Methods

Results

Conclusion