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稳定的埃及芝麻(L.) cv. 苏赫 1 的植物内转化系统。

Stable In-Planta Transformation System For Egyptian Sesame ( L.) cv. Sohag 1.

机构信息

Dept. of Gene transfer, Oil Crops Biotechnology Lab, Agricultural Genetic Engineering Research Institute (AGERI), Agriculture Research Center (ARC), Giza 12619, Egypt.

出版信息

GM Crops Food. 2023 Dec 31;14(1):21-31. doi: 10.1080/21645698.2022.2150041.

Abstract

Sesame ( 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 transformation method. Leaves and flowers excised from the T plants at different developmental stages were PCR screened, and 61/93 seedlings were found to be PCR positive. T 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 T 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 -contaminant.

摘要

芝麻(Sesame)是一种重要的油料作物,也是人类最早认识的油用作物之一。芝麻具有优异的营养和治疗特性;它富含重要的脂肪酸、蛋白质、纤维和重要矿物质。不同基因型的含油率有所不同,但一般占种子干重的 50%以上。为了满足全球对植物油生产不断增长的需求,在全球新开垦地区扩大油作物的种植至关重要。分子育种是品种改良的一种快捷方法,但需要高效的转基因技术。已发表的芝麻转化方法高度特定于属,繁琐,并且涉及制备和测试不同的培养基和外植体。我们使用一种稳定、非侵入性和强大的转化方法生产了转基因芝麻植物。从不同发育阶段的 T 植物上切下的叶片和花朵进行了 PCR 筛选,发现 61/93 株幼苗为 PCR 阳性。来自两条线的 T 种子在生物安全温室设施中发芽,并使用 PCR、草丁膦叶画和喷洒完全成熟的植物(0.02 mg/l)进行草丁膦除草剂喷雾进行筛选;非转基因分离体和对照非转基因植物受到严重损害,最终死亡,而转基因植物不受 Basta 喷雾的影响。T1 植物的 RT-PCR 表明,T 后代中存在 Bar 转录本。此外,使用 NPTII 引物的 RT-PCR 未在转基因芝麻植物中产生任何扩增(NPTII 存在于载体中,但不存在于 T-DNA 区域),表明转基因芝麻植物未受到污染。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/6f1e/9851244/91a65e02546f/KGMC_A_2150041_F0001_OC.jpg

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