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用于高效介导转化的大豆((L.) Merr.)杂交种的创制与评估

Generation and Assessment of Soybean ( (L.) Merr.) Hybrids for High-Efficiency -Mediated Transformation.

作者信息

Khan Muhammad Waqar, Shaheen Aaqib, Zhang Xuebin, Zhang Junli, Dewir Yaser Hassan, Magyar-Tábori Katalin

机构信息

State Key Laboratory of Crop Stress Adaptation and Improvement, Henan Joint International Laboratory for Crop Multi-Omics Research, School of Life Sciences, Henan University, Jinming Road, Kaifeng 475004, China.

State Key Laboratory of Crop Stress Adaptation and Improvement, College of Agriculture, Henan University, Kaifeng 475004, China.

出版信息

Life (Basel). 2024 Dec 12;14(12):1649. doi: 10.3390/life14121649.

Abstract

The -mediated technique is widely employed for soybean transformation, but the efficiency of this method is still relatively modest, in which multiple factors are involved. Numerous chemical and physiological cues from host plants are needed for attraction and subsequent T-DNA integration into the plant genome. Susceptible genotypes may permit this attachment and integration, and the agronomically superior genotypes with susceptibility to would play an important role in increasing transformation efficiency. In this study, we aimed to elevate the -mediated transformation efficiency of soybean by integrating susceptibility alleles from William82 and flavonoids accumulating alleles from LX genotypes in the same soybean line. The crossing was made between LX () and William82 () soybean by hand pollination. Expectedly, the resulting hybrid soybean progenies inherited susceptibility traits and high flavonoid contents (i.e., genistein, genistin, apigenin, naringenin, quercetin, and cinnamic acid) essential for potential plant-pathogen interaction. Furthermore, the progenies and susceptible William82 soybean were subjected to transformation using (GV3101) harboring the GmUbi-3XFlag-35S-GFP and reassembled GmUbi3XFlag-35S-GFP: GUS vectors during separate events. Important transformation-related traits like shoot induction and shoot regeneration ability were also significantly improved in progenies. The progenies designated as ZX-3 exhibited superiority over the William82 parental line in all three traits, i.e., shoot induction, regeneration, and -mediated transformation. The transient transformation efficiency of the ZX-16 line was remarkably higher when half-cotyledon explants were wounded and transformed with harboring GUS assembly vector and then co-cultivated on MS medium supplemented with 2 mg/L spermidine, 0.3 g/L GA3, 0.3 mg/L kinetin, and 1.3 mg/L 6-benzylaminopurine. In addition, the shoot elongation was also higher than that of William82 after two weeks of culture on the shoot induction medium. The newly generated soybeans have the potential to serve as a valuable source for high transgene production and represent a promising avenue for future soybean varietal development.

摘要

农杆菌介导的技术被广泛用于大豆转化,但该方法的效率仍然相对较低,其中涉及多个因素。宿主植物需要众多化学和生理信号来吸引并随后将T-DNA整合到植物基因组中。易感基因型可能允许这种附着和整合,而对农杆菌敏感的农艺学上优良的基因型在提高转化效率方面将发挥重要作用。在本研究中,我们旨在通过在同一大豆品系中整合来自威廉姆斯82的易感等位基因和来自LX基因型的黄酮类化合物积累等位基因,提高农杆菌介导的大豆转化效率。通过人工授粉使LX(♀)和威廉姆斯82(♂)大豆杂交。不出所料,得到的杂交大豆后代继承了潜在植物-病原体相互作用所必需的易感性状和高黄酮类化合物含量(即染料木黄酮、染料木苷、芹菜素、柚皮苷、槲皮素和肉桂酸)。此外,在不同的实验中,使用携带GmUbi-3XFlag-35S-GFP和重组GmUbi3XFlag-35S-GFP:GUS载体的农杆菌(GV3101)对后代和易感的威廉姆斯82大豆进行转化。后代中与转化相关的重要性状如芽诱导和芽再生能力也有显著提高。命名为ZX-3的后代在芽诱导、再生和农杆菌介导的转化这三个性状上均表现出优于威廉姆斯82亲本系。当用携带GUS组装载体的农杆菌对半子叶外植体进行创伤和转化,然后在添加2mg/L亚精胺、0.3g/L GA3、0.3mg/L激动素和1.3mg/L 6-苄基腺嘌呤的MS培养基上共培养时,ZX-16品系的瞬时转化效率显著更高。此外,在芽诱导培养基上培养两周后,芽伸长也高于威廉姆斯82。新培育的大豆有潜力成为高转基因产量的宝贵来源,并代表了未来大豆品种发展的一条有前景的途径。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/19c5/11676979/f35cccf62271/life-14-01649-g001.jpg

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