Application of uniconazole in improving the high-throughput genetic transformation efficiency in maize

IF 4.2 2区生物学 Q2 BIOCHEMISTRY & MOLECULAR BIOLOGY Plant Science Pub Date : 2024-09-28 DOI:10.1016/j.plantsci.2024.112270

Shengnan Liu , Jihui Qiao , Shuaisong Zhang , Minhui Lu , Yongqing Yang , Jinsheng Lai , Yan Guo , Yunlu Shi

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Abstract

Agrobacterium-mediated genetic transformation is the most effective and widely used delivery system for candidate genes and genome editors in maize, which is an important crop with the largest planting area and the highest yield. Here, we used gibberellin synthesis inhibitor, uniconazole, to enhance the stem strength of regenerated plantlets resulting in a significantly increase from 11.6 % to 18.2 % in the percentage of regenerated plantlets, and the transformation frequency was also improved from 9.4 % to 15.6 % in the test experiments. The physiological condition of immature embryo is greatly affected by ear source, season and insect pests, while it can cause significant fluctuations in the transformation frequency. Our optimization works at the differentiation subculture stage, avoiding the impact on the physiological condition of immature embryo. So, it can be applicated to high-throughput genetic transformation in different seasons and different ear sources throughout the year. The productive experiment results indicated that the annual average transformation frequency significantly improved from 2.76 % to 7.14 % (approximately 2.6 folds improvement), and the tissue culture cycle was shortened from 115 days to 106 days by using optimized system. Our optimized genetic transformation system opens avenues for maize improvement based on transgenic and genome editing technology.

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在提高玉米高通量遗传转化效率中应用联苯苄唑。

玉米是种植面积最大、产量最高的重要作物，农杆菌介导的遗传转化是玉米候选基因和基因组编辑最有效、应用最广泛的传递系统。在这里，我们使用赤霉素合成抑制剂--烯效唑来增强再生小苗的茎干强度，结果再生小苗的比例从 11.6% 显著提高到 18.2%，转化频率也从 9.4% 提高到 15.6%。未成熟胚的生理状况受穗源、季节和虫害的影响很大，同时也会导致转化频率的大幅波动。我们的优化方案在分化亚培养阶段就能发挥作用，避免了对未成熟胚胎生理状况的影响。因此，它可以应用于全年不同季节和不同耳源的高通量遗传转化。生产性实验结果表明，使用优化系统后，年平均转化率从 2.76% 显著提高到 7.14%（约提高 2.6 倍），组织培养周期从 115 天缩短到 106 天。我们的优化遗传转化系统为基于转基因和基因组编辑技术的玉米改良开辟了道路。

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来源期刊

Plant Science 生物-生化与分子生物学

CiteScore

9.10

自引率

1.90%

发文量

322

审稿时长

33 days

期刊介绍： Plant Science will publish in the minimum of time, research manuscripts as well as commissioned reviews and commentaries recommended by its referees in all areas of experimental plant biology with emphasis in the broad areas of genomics, proteomics, biochemistry (including enzymology), physiology, cell biology, development, genetics, functional plant breeding, systems biology and the interaction of plants with the environment. Manuscripts for full consideration should be written concisely and essentially as a final report. The main criterion for publication is that the manuscript must contain original and significant insights that lead to a better understanding of fundamental plant biology. Papers centering on plant cell culture should be of interest to a wide audience and methods employed result in a substantial improvement over existing established techniques and approaches. Methods papers are welcome only when the technique(s) described is novel or provides a major advancement of established protocols.