Production of transformed bitter gourd plants through in vitro regeneration is a laborious practice, which may also result in somaclonal variations. Thereby, in the current study, we have established a less tedious in planta protocol for more efficient genetic transformation of bitter gourd var. NBGH-470 (Apurva) using the seed as a target material. Bitter gourd seeds were transformed by Agrobacterium tumefaciens strain LBA4404 bearing the pCAMBIA1301 binary vector, and the transformed plants were selected against hygromycin B. The putatively transformed bitter gourd (T_o) plants were examined by GUS assay. Two parameters, including vacuum infiltration and sonication improving the in planta transformation, have been investigated in the present work. Amongst several time durations examined, the highest transformation rate (37%) was achieved upon subjecting the pre-cultured bitter gourd seeds to sonication for 15 min, followed by vacuum infiltration for 6 min in LBA4404 culture suspension. The transformed bitter gourd (T₁) plants were selected against hygromycin B, and the transgene integration was evaluated by polymerase chain reaction (PCR), Southern hybridization, and reverse transcriptase PCR (RT-PCR). The developed protocol in the current study is suitable to genetically engineer the bitter gourd plants with disease and pest-resistant traits.

通过体外再生生产转化的苦瓜植物是费力的实践，这也可能导致体细胞克隆变异。因此，在当前的研究中，我们已经建立了一个不太繁琐的植物方案，以更有效地进行苦瓜变种的遗传转化。NBGH-470（Apurva），使用种子作为目标材料。苦瓜种子通过转化根癌农杆菌菌株LBA4404轴承质粒pCAMBIA1301二元载体，并选择对潮霉素B的转化植物推定转化苦瓜（T _ö通过GUS测定法检查植物。在目前的工作中，已经研究了两个参数，包括真空渗透和超声处理以改善植物体内的转化。在所考察的几个时间段中，对预培养的苦瓜种子进行超声处理15分钟，然后在LBA4404培养悬浮液中进行真空浸润6分钟，从而实现了最高转化率（37％）。针对潮霉素B选择转化的苦瓜（T ₁）植物，并通过聚合酶链反应（PCR），Southern杂交和逆转录酶PCR（RT-PCR）评估转基因整合。本研究中开发的方案适用于遗传改造具有疾病和抗虫特性的苦瓜植物。