Direct regeneration of haploid or doubled haploid plantlets in cucumber ( Cucumis sativus L.) through ovary culture
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ORIGINAL ARTICLE
Direct regeneration of haploid or doubled haploid plantlets in cucumber (Cucumis sativus L.) through ovary culture Ying Deng1,2 · Bing Tang2 · Xia Zhou1 · Wenyuan Fu2 · Lian Tao2 · Lu Zhang1 · Jinfeng Chen1 Received: 22 August 2019 / Accepted: 30 April 2020 © Springer Nature B.V. 2020
Abstract This paper reports a simple and effective method of directly producing cucumber plants through unfertilized ovary culture. The paper involves an experiment investigating factorial interactions between TDZ treatment, cold pretreatment, genotypes of cucumber, to improve plant induction. Cold pretreatment was effective in stimulating the ovary. The results showed that cold pretreatment for 4 days, TDZ for 0.06 mg·L−1, the interaction with genotype can be used as an effective strategy to improve the efficiency of gynogenesis. The plant regeneration induction rate was highest (79.3%). In addition, we observed the process of cucumber megasporogenesis and plant regeneration. The plants obtained from ovary culture of cucumber were identified as diploid or haploid by flow cytometry, consistent with the results of chromosome counting. The diploid plants were further identified as pure doubled haploid using simple sequence repeats (SSR). The doubling treatment we used was one of the simplest and most effective methods, completed in a short time (1 h) with a doubling rate of 75%. The acclimation rate for the surviving was 70%. This work provides a basis for promoting haploid breeding in cucumber. Keywords Cucumber (cucumis sativus L.) · Ovary culture · Doubled haploid · Direct regeneration
Key message Communicated by Jose M. Segui-Simarro. Ying Deng and Bing Tang have contributed equally to this work. * Jinfeng Chen [email protected] Ying Deng [email protected] Bing Tang [email protected] Xia Zhou [email protected] Wenyuan Fu [email protected] Lian Tao [email protected] Lu Zhang [email protected] 1
State Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China
Institute of Horticulture, Guizhou Academy of Agricultural Sciences, Guiyang 55006, China
2
An effective technique for the production of double haploids was provided through ovary culture in cucumber (Cucumis sativus L.). More interesting, we observed the entire process of gynogenesis.
Introduction Gynogenesis refers to the process of embryo regeneration from unfertilized female gametophytes in vitro or in vivo (Gémes-Juhász et al. 2002). In contrast to somatic embryogenesis, which results in clonal propagation of a specific genotype, gynogenic plants exhibit the product of meiotic segregation. They have the important characteristic of containing only one set of chromosomes and are therefore haploid plants (Maraschin et al. 2005). Homozygous double haploids obtained by colchicine-induced or spontaneous doubling are used in breeding. The process of inducing female gametophytes to form embryos is difficult to sustainable observe, because the embryo sac is embedded in somatic cell tissues, making it di
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