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Successful application of microspore culture in pakchoi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee) for hybrid breeding

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Microspore embryogenesis is an effective method of obtaining double haploid (DH) lines in only 1 year. However, the microspore embryogenesis protocol was not efficient in pakchoi. This study aimed to establish an effective microspore culture protocol in pakchoi for hybrid breeding. The embryos were obtained from three genotypes (18SY01, 18SY02, 18SY03), but the frequency of microspore embryogenesis was significantly different. Globular embryos from three genotypes were placed into a rotary shaker (50 r/min, 25 ℃) for further culture to improve microspore embryogenesis and plantlet regeneration without callus development. Shake culture not only increased the frequency of cotyledonary embryos but also accelerated microspore embryogenesis in the NLN-13 liquid medium. Moreover, the doubled haploid rates of regenerated plants for the three genotypes were above 50%. The morphological characters and plot yield of DH lines were identified, and there were significant differences between them. According to the measurement of the self-compatibility index, all the DH lines were self-incompatible. Furthermore, the hybrid combination was prepared with the selected DH lines and the pakchoi genic male sterile line GMS010 to develop excellent hybrids. This work contributes to accelerating the application of microspore embryogenesis and supplying the DH lines in pakchoi hybrid breeding.

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Doubled haploid


1-Naphthylacetic acid


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This work was supported by the National Natural Science Foundation of China (31972404), China Agriculture Research System (CARS-23), and Science Study Foundation of Liaoning (LJKZ0640).

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Correspondence to Yun Zhang.

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Fang, S., Ma, Y., Liu, Z. et al. Successful application of microspore culture in pakchoi (Brassica campestris L. ssp. chinensis Makino var. communis Tsen et Lee) for hybrid breeding. Protoplasma 260, 545–555 (2023).

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