Beijing Academy of Agriculture and Forestry Sciences, Key Laboratory of Urban Agriculture (North China), Ministry of Agriculture and Rural Affairs, Beijing 100097, China.
College of Forestry and Grassland Science, Jilin Agricultural University, Changchun 130118, China.
Int J Mol Sci. 2023 Oct 18;24(20):15304. doi: 10.3390/ijms242015304.
Lily ( spp.) is a popular ornamental plant. Traditional genetic transformation methods have low efficiency in lily, thus development of a high-efficiency genetic transformation system is important. In this study, a novel transient transformation method involving pollen magnetofection was established and optimized pollen viability, and exogenous gene expression in magnetofected pollen and that of different germplasm were assessed. The highest germination percentage of pollen was 85.73% in medium containing 100 g/L sucrose, 61.5 mg/L HBO, and 91.5 mg/L CaCl. A 1:4 ratio of nanomagnetic beads to DNA plasmid and transformation time of 0.5 h realized the highest transformation efficiency (88.32%). The GFP activity in transformed pollen averaged 69.66%, while that of the control pollen was 0.00%. In contrast to the control, transgenic seedlings obtained by pollination with magnetofected pollen showed strong positive GUS activity with 56.34% transformation efficiency. Among the lily germplasm tested, 'Sweet Surrender' and had the highest transformation efficiency (85.80% and 54.47%), whereas var. was not successfully transformed. Transformation efficiency was positively correlated with pollen equatorial diameter and negatively correlated with polar axis/equatorial diameter ratio. The results suggest that pollen magnetofection-mediated transformation can be applied in but might have species or cultivar specificity.
百合（ spp.）是一种受欢迎的观赏植物。传统的遗传转化方法在百合中的效率较低，因此开发高效的遗传转化系统很重要。本研究建立了一种新的涉及花粉磁转染的瞬时转化方法，并优化了花粉活力以及磁转染花粉中外源基因的表达和不同种质的表达。在含有 100 g/L 蔗糖、61.5 mg/L HBO 和 91.5 mg/L CaCl 的培养基中，花粉的最高萌发率为 85.73%。纳米磁珠与 DNA 质粒的比例为 1:4，转化时间为 0.5 h 时，转化效率最高（88.32%）。转化花粉中的 GFP 活性平均为 69.66%，而对照花粉为 0.00%。与对照相比，用磁转染花粉授粉获得的转基因幼苗表现出强烈的阳性 GUS 活性，转化效率为 56.34%。在所测试的百合种质中，'甜蜜投降'和 具有最高的转化效率（85.80%和 54.47%），而 var. 则未成功转化。转化效率与花粉赤道直径呈正相关，与极轴/赤道直径比呈负相关。结果表明，花粉磁转染介导的转化可以应用于 ，但可能具有物种或品种特异性。
