Open Access

ARTICLE

Optimization of Callus Induction Conditions from Immature Embryos in Maize and Plant Regeneration

Peng Jiao^1,#, Ruiqi Ma^2,#, Zhuo Qi¹, Zhenzhong Jiang¹, Siyan Liu¹, Jing Qu³, Shuyan Guan^1,*, Yiyong Ma^3,*

1 College of Life Sciences, Jilin Agricultural University, Changchun, 130118, China
2 Northeast Normal University, Environmental School, Changchun, 130118, China
3 College of Agronomy, Jilin Agricultural University, Changchun, 130118, China
# These authors contributed equally to this work

* Corresponding Author: Shuyan Guan. Email: ; Yiyong Ma. Email:

Phyton-International Journal of Experimental Botany 2020, 89(1), 121-130. https://doi.org/10.32604/phyton.2020.07980

Received 17 July 2019; Accepted 17 October 2019; Issue published 01 March 2020

Abstract

This research uses the immature embryos of inbred maize lines (GSH9901, Hi01, Hi02, and Chang 7-2) as receptor materials to establish the callus induction system. These inbred lines provide the receptor materials for the genetic regeneration of maize and the verification of the genetic functions of maize. The factor experiment and orthogonal experiments were used to investigate the impacts of different genotypes, immature embryo size, shield orientation, 2, 4-D concentration, proline concentration, and folic acid concentration on the induction rate of embryogenic callus tissue. A sensitivity experiment testing glyphosate (Bar) and an antibiotic (Cefotaxime sodium) were also conducted. The results indicate that the immature embryos of inbred maize line GSH9901 were the most effective for callus tissue induction, and the immature embryos with a length of 1.6-2.0 mm produce the best result. The upward shield face is more successful for the formation of induced callus. Using orthogonal analysis, we found that the optimal combination for the induction system was A₃ (2,4-D concentration 0.25 mg mL^-1 ), B₁C₃ (proline concentration 0.8 mg mL^-1 ), and D₂ (folate Concentration 0.5 mg mL^-1) and the induction rate reached 84%. We found that cold storage at 4 °C for 1 d is more conducive for the formation of embryogenic callus than the other treatments tested. The sensitivity experiment for callus tissue screening revealed the critical concentration of glyphosate to be 10 mg ml^-1 , and the critical concentration of antibiotic is 250 mg ml^-1 . Using this combination of glyphosate and antibiotic resulted in regenerated plants. This study established the optimal conditions for immature embryo callus tissue induction in maize.

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Keywords

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