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Acta Horticulturae Sinica ›› 2021, Vol. 48 ›› Issue (11): 2197-2210.doi: 10.16420/j.issn.0513-353x.2020-1078

• Original article • Previous Articles     Next Articles

High-throughput Screening of Transmission Rate for Ogura CMS Fertility Restorer Gene Rfo by HRM Technique in Cabbage

REN Wenjing, YU Hailong, CHEN Li, ZHANG Bin, CHEN Wendi, FANG Zhiyuan, YANG Limei, ZHUANG Mu, LÜ Honghao, WANG Yong, JI Jialei, ZHANG Yangyong()   

  1. Institute of Vegetables and Flowers,Chinese Academy of Agricultural Sciences,Beijing 100081,China
  • Received:2021-02-25 Revised:2021-08-20 Published:2021-12-02

Abstract:

In order to create the cabbage Ogura CMS fertility restorer,backcrosses were conducted after distant hybridization between Brassica oleracea L. var. alboglabra and Brassica napus. BC4 and BC5 generations were developed. In this study,a Rfo-specific InDel molecular marker Rfo-11F/Rfo-11R suitable for high-resolution melting curve(HRM)analysis was designed and developed based on the Rfo gene coding sequence and promoter sequence. This marker was used for Rfo genotyping of BC4 and BC5 generations of the distant hybridization. A total of 41 progeny in BC4 generation with Rfo gene were successfully selected with the average Rfo transmission rate of 0.51% and pollen viability of 81%. Five key Rfo-positive individuals were selected for further backcross. Cytological observation showed that the average proportion of normal pairing and segregating of pollen mother cells in BC4 generation were 84% and 69%,respectively. After further backcrossing,117 Rfo-positive individuals were obtained from 15 408 BC5 backcrossing progeny,and the Rfo gene transmission rate was 0.75%. The mean value of pollen viability was 83%,and the proportion of pollen mother cells with normal pairing and segregating chromosomes were 81% and 84%,respectively. After five successive generations of backcrossing,all the fertility-restored cabbage individuals returned to normal diploid level and exhibited fertility-restored,which was consistent with the molecular marker identification results. This indicated that the fast,accurate and efficient high-throughput genotyping system for Ogura CMS fertility restorer gene Rfo,which was constructed based on the developed InDel marker Rfo-11F/Rfo-11R,can be used for further screen of backcrossing progeny and great improvement of the breeding efficiency.

Key words: cabbage, high-resolution melting(HRM), Ogura CMS, fertility restorer gene, high-throughput

CLC Number: