ABSTRACT

Sesame (Sesamum indicum L.) is an important oil crop and one of the oldest-known oil crops to humankind. Sesame has excellent nutritional and therapeutic properties; it is rich in important fatty acids, protein, fiber, and vital minerals. Oil percentage varies among different genotypes but generally accounts for more than 50% of the seed’s dry weight. To meet the increasing demand for vegetable oil production worldwide, expanding the cultivation of oil crops in newly reclaimed areas worldwide is essential. Molecular breeding is an expeditious approach for varietal improvement but requires efficient transgenesis. Published sesame transformation methods are highly genus-specific, tedious, and involve preparing and testing different media and explants. We produced transgenic sesame plants using a stable, noninvasive, and robust Agrobacterium tumefaciens transformation method. Leaves and flowers excised from the T₀ plants at different developmental stages were PCR screened, and 61/93 seedlings were found to be PCR positive. T₁ seeds resulting from two lines were germinated in a biocontainment greenhouse facility and screened using PCR, basta leaf painting, and spraying fully matured plants with basta herbicide (0.02 mg/l); non-transgenic segregants and control non-transgenic plants were severely damaged, and eventually died, while transgenic plants were not affected by the Basta spraying. RT-PCR on T1 plants indicated the presence of Bar transcripts in T₁ progeny. Furthermore, RT-PCR using NPTII primers did not result in any amplification in transgenic sesame plants (NPTII is present in the vector but not in the T-DNA region) indicating that the transgenic sesame plants were not an Agrobacterium-contaminant.

摘要

芝麻 ( Sesamum indicumL.) 是一种重要的油料作物，也是人类已知最古老的油料作物之一。芝麻具有极好的营养和治疗特性；它富含重要的脂肪酸、蛋白质、纤维和重要矿物质。不同基因型的油百分比不同，但通常占种子干重的 50% 以上。为了满足全球对植物油生产日益增长的需求，在全球新开垦地区扩大油料作物种植至关重要。分子育种是品种改良的快速方法，但需要有效的转基因。已发布的芝麻转化方法具有高度属特异性、繁琐，并且涉及准备和测试不同的培养基和外植体。我们使用稳定、无创且稳健的方法生产转基因芝麻植物根癌农杆菌转化方法。_{从不同发育阶段的T 0}植株切下的叶和花进行PCR筛选，发现61/93的幼苗为PCR阳性。来自两个品系的T ₁种子在生物防护温室设施中发芽，并使用 PCR、basta 叶涂和用 basta 除草剂（0.02 mg/l）喷洒完全成熟的植物进行筛选；非转基因分离子和对照非转基因植物受到严重破坏，并最终死亡，而转基因植物不受巴斯塔喷洒的影响。_{T1 植物上的 RT-PCR 表明 T 1}中存在 Bar 转录物后代。此外，使用 NPTII 引物的 RT-PCR 没有导致转基因芝麻植物中的任何扩增（NPTII 存在于载体中但不存在于 T-DNA 区域中）表明转基因芝麻植物不是农杆菌污染物。