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Development of a new codominant CAPS marker for sex genotype identification in asparagus

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Abstract

Garden asparagus (Asparagus officinalis L.) is a dioecious species, with male [XY] and female [XX] individuals. Since male individuals are preferred over females for agricultural production, all-male cultivars have agronomic advantages over mixed-sex cultivars. To produce an all-male cultivar, it is important to obtain a supermale [YY]. Given their morphological similarities, males and supermales are usually distinguished by genetic analysis. To reduce the time required for asparagus breeding, various dominant and codominant male-specific DNA markers have been developed to date. Although two sex-linked codominant markers (Asp2-SP6 and RM17) have been reported previously, their ability to distinguish the sex in A. officinalis and other dioecious Asparagus species has not been evaluated extensively. Therefore, we analyzed the application of these markers to A. officinalis, purple asparagus cultivars and other dioecious Asparagus species in this study. Our results showed that the capacity of these markers to distinguish the sex in A. officinalis and purple asparagus cultivars was limited, thus we developed a new CAPS marker (SSM01), based on the sequence around the RM17 marker. Since SSM01 amplify ca. 470 bp fragment and only Y-specific fragment was digested into ca. 270/200 bp by XspI, this marker can discriminate between females, males and supermales. SSM01 is applicable for discrimination between X- and Y-specific sequences in various A. officinalis and purple asparagus cultivars, also in three dioecious Asparagus species.

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All data generated or analysed during this study are included in this published article and its supplementary information files.

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Acknowledgements

We thank Prof. Takahiro Sonoda (Rakuno Gakuen University, Japan) for providing plant materials of ‘RG murasakishikibu Luce’ and ‘RG murasakishikibu First’. This work was supported by JSPS KAKENHI (Grant Numbers: 18H02192 and 18K19197) and by the research program on development of innovative technology Grants (JPJ007097) from the Project of the Bio-oriented Technology Research Advancement Institution (BRAIN).

Funding

This work was supported by JSPS KAKENHI (Grant Numbers: 18H02192 and 18K19197) and by the research program on development of innovative technology Grants (JPJ007097) from the Project of the Bio-oriented Technology Research Advancement Institution (BRAIN).

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Authors and Affiliations

  1. Graduate School of Life Sciences, Tohoku University, Katahira 2-1-1, Aoba-ku, Sendai, 980-8577, Japan

    Mako Akahori & Akira Kanno

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  1. Mako Akahori
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  2. Akira Kanno
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All authors contributed to the study conception and design. Material preparation, data collection and analysis were performed by MA. The manuscript was written by both authors.

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Correspondence to Akira Kanno.

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Supplementary Information

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10681_2022_3029_MOESM1_ESM.pdf

Supplementary file1 (PDF 348 kb)—PCR analysis of A. officinalis and purple asparagus cultivars using the Asp2-SP6 marker. a Six F1 individuals generated by self-fertilizing the andromonoecious line ‘Gijnlim’ [XY]. Lanes 1, 2: females [XX]; 3, 4: males [XY]; 5, 6: supermales [YY]. b Four A. officinalis cultivars. Lanes 1–3, 7–9, 16–18: females [XX]; 4–6, 10–15, 19-21: males [XY]. c Two purple asparagus cultivars. Lanes 1–3, 7–9: females; 4–6, 10–12: males. In a–c, the top image shows fragments amplified by Asp2-SP6 primers, and the bottom image shows Asp2-SP6 amplicons digested by MseI. Lane M: 100 bp ladder. Arrows indicate 500 bp

10681_2022_3029_MOESM2_ESM.pdf

Supplementary file2 (PDF 306 kb)—PCR analysis of asparagus cultivars using the RM17 marker. Black triangles indicate X- and Y-specific fragments (189 and 215 bp, respectively). a Six F1 individuals derived by self-fertilizing the andromonoecious line ‘Gijnlim’ [XY]. Lanes 1, 2: females [XX]; 3, 4: males [XY]; 5, 6: supermales [YY]. b Four A. officinalis cultivars. Lanes 1–3, 7–9, 13–15, females [XX]; 4–6, 10–12, 16–21, males [XY]. c Two purple asparagus cultivars. Lanes 1–3, 7–9: females; 4–6, 10–12: males

10681_2022_3029_MOESM3_ESM.pdf

Supplementary file3 (PDF 593 kb)—Alignment of X- and Y-specific sequences around the SSM01 marker in A. officinalis ‘Mary Washington 500 W’ (MW), ‘Gijnlim’ (GM) and purple asparagus cultivars ‘Purple Passion’ (Pas) and ‘Pacific Purple’ (Pfc). Dotted arrows indicate the annealing sites of RM17su-fw and RM17su-rv primers, and solid arrows indicate the annealing sites of SSM01-fw and SSM01-rv primers. Black box indicates the Y-specific XspI site. Red squares indicate Y-specific polymorphisms

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Akahori, M., Kanno, A. Development of a new codominant CAPS marker for sex genotype identification in asparagus. Euphytica 218, 75 (2022). https://doi.org/10.1007/s10681-022-03029-5

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