Naqvi Rubab Z, Asif Muhammad, Saeed Muhammad, Asad Shaheen, Khatoon Asia, Amin Imran, Mukhtar Zahid, Bashir Aftab, Mansoor Shahid
Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic EngineeringFaisalabad, Pakistan; Pakistan Institute of Engineering and Applied SciencesNilore, Pakistan.
Agricultural Biotechnology Division, National Institute for Biotechnology and Genetic Engineering Faisalabad, Pakistan.
Front Plant Sci. 2017 Jan 24;8:55. doi: 10.3389/fpls.2017.00055. eCollection 2017.
Insect pest complex, cotton leaf curl disease and weeds pose major threat to crop production worldwide, including Pakistan. To address these problems, in the present study a triple gene construct harboring , , and cassettes has been developed for plant specifically in cotton transformation against lepidopteron insect-pests and weeds. (tobacco) was used as a model system for characterization of this triple gene construct. The construct has been assembled in plant expression vector and transformed in In six transgenic tobacco lines the integration of - in tobacco genome was checked by PCR, while successful protein expression of all the three genes was confirmed through immunostrip assay. Efficacy of and was evaluated through insect bioassay using armyworm (). These transgenic tobacco plants showed significant insect mortality as compared to control plants during insect bioassay. Three out of six tested transgenic lines L3, L5, and L9 exhibited 100% mortality of armyworm, while three other lines L1, L10, and L7 showed 86, 80, and 40% mortality, respectively. This construct can readily be used with confidence to transform cotton and other crops for the development of insect resistant and herbicide tolerant transgenic plants. The transgenic crop plants developed using this triple gene construct will provide an excellent germplasm resource for the breeders to improve their efficiency in developing stable homozygous lines as all the three genes being in a single T-DNA border will inherit together.
害虫复合体、棉花卷叶病和杂草对包括巴基斯坦在内的全球作物生产构成重大威胁。为解决这些问题，在本研究中，已开发出一种包含 、 和 盒的三基因构建体，专门用于棉花转化，以对抗鳞翅目害虫和杂草。 （烟草）被用作表征该三基因构建体的模型系统。该构建体已组装到植物表达载体中并转化到 中。在六个转基因烟草品系中，通过PCR检测了 - 在烟草基因组中的整合情况，同时通过免疫条带试验确认了所有三个基因的成功蛋白表达。通过使用粘虫（ ）进行昆虫生物测定来评估 和 的功效。在昆虫生物测定期间，与对照植物相比，这些转基因烟草植物表现出显著的昆虫死亡率。六个测试的转基因品系中的三个，即L3、L5和L9，表现出粘虫100%的死亡率，而其他三个品系L1、L10和L7分别表现出86%、80%和40%的死亡率。该构建体可放心地用于转化棉花和其他作物，以培育抗虫和耐除草剂的转基因植物。使用这种三基因构建体培育的转基因作物将为育种者提供优秀的种质资源，以提高他们开发稳定纯合品系的效率，因为所有三个基因位于单个T-DNA边界内将一起遗传。
