UMR-BFP-1332, INRAE-Bordeaux, Bordeaux-UniversityII, 71 Avenue Bourleaux, 33883 Villenave d'Ornon, France.
USDA-ARS Fruit Station, 2217 Wiltshire Road, Kearneysville, WV 25430, USA.
Genes (Basel). 2021 May 27;12(6):816. doi: 10.3390/genes12060816.
Our goal was to target silencing of the coat protein ( CP) gene independently expressed in plants. Clone C-2 is a transgenic plum expressing CP. We introduced and verified, in planta, the effects of the inverse repeat of CP sequence split by a hairpin (IRSH) that was characterized in the HoneySweet plum. The IRSH construct was driven by two CaMV35S promoter sequences flanking the CP sequence and had been introduced into C1738 plum. To determine if this structure was enough to induce silencing, cross-hybridization was made with the C1738 clone and the CP expressing but -susceptible C2 clone. In total, 4 out of 63 clones were silenced. While introduction of the IRSH is reduced due to the heterozygous character in C1738 plum, the silencing induced by the IRSH CP is robust. Extensive studies, in greenhouse containment, demonstrated that the genetic resource of C1738 clone can silence the CP production. In addition, these were verified through the virus transgene pyramiding in the BO70146 BlueByrd cv. plum that successfully produced resistant BlueByrd BO70146 × C1738 (HybC1738) hybrid plums.
我们的目标是独立地靶向植物中表达的外壳蛋白（CP）基因的沉默。克隆 C-2 是一种表达 CP 的转基因李树。我们在体内引入并验证了在 HoneySweet 李树上具有特征的 CP 序列通过发夹（IRSH）分裂的反向重复的效果。IRSH 构建体由两个 CaMV35S 启动子序列侧翼的 CP 序列驱动，并已被引入 C1738 李树。为了确定这种结构是否足以诱导沉默，与 C1738 克隆和表达但易感的 C2 克隆进行了交叉杂交。总共，63 个克隆中有 4 个被沉默。虽然由于 C1738 李树的杂合子特性导致 IRSH 的引入减少，但 IRSH-CP 诱导的沉默是强大的。在温室隔离的广泛研究中，证明了 C1738 克隆的遗传资源可以沉默 CP 的产生。此外，通过在 BO70146 BlueByrd cv 中进行病毒转基因叠加进行了验证。 plum 成功生产了抗性 BlueByrd BO70146×C1738（HybC1738）杂交李树。
