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. 2025 Sep 9:2025:10.17912/micropub.biology.001772.
doi: 10.17912/micropub.biology.001772. eCollection 2025.

A simple protocol for producing axenic seeds of Sorghum bicolor

Beatrice Bock  1   2 , Jack Scherer  1 , Faith Parrish  1 , Julia Burnside  1 , Callum Rohrer  1   2 , Catherine Gehring  1   2
Affiliations

A simple protocol for producing axenic seeds of Sorghum bicolor

Beatrice Bock et al. MicroPubl Biol. .

Abstract

Microbes within seeds can confound research on microbial colonization, symbiosis, and pathogenesis. Sterilization of both external and internal seed tissues is therefore essential in certain experiments, but the method must also preserve seed viability. Here, we present a reliable and simple protocol for sterilizing Sorghum bicolor seeds by submerging them in 95% ethanol for 2 minutes followed by 3.75% sodium hypochlorite for 20 minutes. This approach yielded a low contamination rate (2 out of 95 seeds) and a robust median germination rate (63%). Its simplicity, cost-effectiveness, and accessibility make it a practical option for experiments requiring axenic seeds.

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Conflict of interest statement

The authors declare that there are no conflicts of interest present.

Figures

Figure 1. <b> Prevalence of germination and contamination among <i>Sorghum bicolor</i> seeds treated with the described sterilization protocol </b>
Figure 1. Prevalence of germination and contamination among Sorghum bicolor seeds treated with the described sterilization protocol
Each X symbol represents a group’s observed proportion of germinated or contaminated seeds. Each group consisted of 7-8 seeds treated at the same time with the described method. Of the 95 seeds tested, only two showed contamination by culturable microbes. Both seeds came from the same group. Boxplot elements include a horizontal line at the median (contamination = 0, germination = 0.63), a box showing the interquartile range (IQR), whiskers extending to values within 1.5 × IQR, and black circles marking outliers.
See this image and copyright information in PMC

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