The RUBY reporter enables efficient haploid identification in maize and tomato.
The RUBY reporter enables efficient haploid identification in maize and tomato.
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DOI：
10.1111/pbi.14071
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发表时间：
2023-08
影响因子：
13.8
通讯作者：
Chen, Shaojiang
中科院分区：
文献类型：
--
作者：
Wang, Dong;Zhong, Yu;Feng, Bin;Qi, Xiaolong;Yan, Tongzheng;Liu, Jinchu;Guo, Shuwei;Wang, Yuwen;Liu, Zongkai;Cheng, Dehe;Zhang, Yuling;Shi, Yunlu;Zhang, Shuaisong;Pan, RuXue;Liu, Chenxu;Chen, Shaojiang
体内单倍体诱导已经从玉米扩展到单子叶植物，如水稻、小麦、谷子和双子叶植物，如番茄、油菜籽、烟草和卷心菜。单倍体的准确鉴定是加倍单倍体技术的关键步骤，一个有效的鉴定标记是非常关键的。在玉米单倍体鉴定中，R1-NJ是一种广泛使用的视觉标记。RFP和EGFP已被证明是鉴定单倍体的可行方法。然而，这些方法要么限于特定的物种，要么需要特定的设备。它仍然缺乏一种有效的视觉标记，可以在不同的作物物种中使用。在本研究中，我们将甜菜碱生物合成系统Ruby Report引入玉米和番茄单倍体诱导子中，作为单倍体鉴定的新标记。结果表明，在授粉后10 天，红宝石基因的表达可导致玉米胚中甜菜碱的深度着色，并使未成熟单倍体胚胎鉴定的准确率达到100%。在番茄上的进一步研究表明，新的标记导致胚根中的深红色色素沉着，单倍体可以容易和准确地识别。结果表明，Ruby报告基因是一种不依赖于背景的、高效的单倍体鉴定标记，在不同作物种的加倍单倍体育种中具有广阔的应用前景。
In vivo haploid induction has been extended from maize to monocotyledonous plants like rice, wheat, millet and dicotyledonous plants such as tomato, rapeseed, tobacco and cabbage. Accurate identification of haploids is a crucial step of doubled haploid technology, where a useful identification marker is very pivotal. R1‐nj is an extensively used visual marker for haploid identification in maize. RFP and eGFP have been shown to be feasible in identifying haploid. However, these methods are either limited to specific species, or require specific equipment. It still lacks an efficient visual marker that is practical across different crop species. In this study, we introduced the RUBY reporter, a betalain biosynthesis system, into maize and tomato haploid inducers as a new marker for haploid identification. Results showed that expression of RUBY could result in deep betalain pigmentation in maize embryos as early as 10 days after pollination, and enabled 100% accuracy of immature haploid embryo identification. Further investigation in tomato revealed that the new marker led to deep red pigmentation in radicles and haploids can be identified easily and accurately. The results demonstrated that the RUBY reporter is a background‐independent and efficient marker for haploid identification and would be promising in doubled haploid breeding across different crop species.
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