Pepper Jordan T, Maheshwari Priti, Eudes François
Agriculture and Agri-food/Agriculture et Agroalimentaire Canada, Lethbridge Research Centre, 54031st Avenue South, Lethbridge, Alberta, T1J 4P4, Canada.
Colloids Surf B Biointerfaces. 2017 Sep 1;157:207-214. doi: 10.1016/j.colsurfb.2017.05.075. Epub 2017 Jun 3.
Microspores are the pre-gametophyte stage of pollen, and have proven to be a successful tissue culture material for the production of doubled haploid plants. Microspore culture has also been used as a platform for the production of transgenic plants. The use of cell-penetrating peptides (CPPs) as transfection agents in microspores has been previously demonstrated, but at low efficiencies. Here, the pH dependent adsorption of the cationic CPP Tat to the sporopollenin surface of the microspore (the exine) has been explored using electrophoretic light scattering (ELS). Furthermore the adsorption of a commercially available polycationic polymer; Luviquat FC-370 (polyquaternium D-16, PQ-D16) was similarly measured using ELS. It was found that a suspension media with a pH less than 7.0 showed an approximately ten fold decrease in the amount of Tat that was required before apparent surface neutralization. This data suggests that a change in the surface chemistry of the microspore occurs in acidic pH conditions, that modulates the binding affinity of the CPP Tat in a non-trivially complex manner.
小孢子是花粉的配子体前期阶段，并且已被证明是用于生产双单倍体植物的一种成功的组织培养材料。小孢子培养也已被用作生产转基因植物的一个平台。此前已证明使用细胞穿透肽（CPPs）作为小孢子中的转染剂，但效率较低。在这里，已使用电泳光散射（ELS）研究了阳离子CPP Tat对小孢子孢粉素表面（外壁）的pH依赖性吸附。此外，使用ELS类似地测量了市售聚阳离子聚合物卢维夸特FC - 370（聚季铵盐D - 16，PQ - D16）的吸附。发现pH小于7.0的悬浮培养基在表面明显中和之前所需的Tat量减少了约十倍。该数据表明，在酸性pH条件下小孢子的表面化学发生了变化，以一种非常复杂的方式调节了CPP Tat的结合亲和力。
