title: Efficient regeneration and improved sonication-assisted Agrobacterium transformation (SAAT) method for Catharanthus roseus.
abstract: Catharanthus roseus is an important medicinal plant known for its pharmacological qualities such as antimicrobial, anticancerous, antifeedant, antisterility, antidiabetic activities. More than 130 bioactive compounds like vinblastine, vindoline and vincristine have been synthesized in this plant. Extensive studies have been carried out for optimization regeneration and transformation protocols. Most of the protocol described are laborious and time-consuming. Due to sophisticated protocol of regeneration and genetic transformation, the production of these bioactive molecules is less and not feasible to be commercialized worldwide. Here we have optimized the efficient protocol for regeneration and transformation to minimize the time scale and enhance the transformation frequency through Agrobacterium and sonication-assisted transformation (SAAT) method. In this study, hypocotyl explants responded best for maximal production of transformed shoots. The callus percentage were recorded 52% with 1.0 mg L-1(BAP) and 0.5 mg L-1(NAA) while 80% shoot percentage obtained with 4.0 mg L-1(BAP) and 0.05 mg L-1(NAA). The microscopic studies revealed that the expression of GFP was clearly localized in leaf tissue of the C. roseus after transformation of pRepGFP0029 construct. Consequently, transformation efficiency was revealed on the basis of GFP localization. The transformation efficiency of SAAT method was 6.0% comparable to 3.5% as conventional method. Further, PCR analysis confirmed the integration of the nptII gene in the transformed plantlets of C. roseus.
authors.affiliation: Sara Alghonaim Research Chair (SRC), Biology Department, College of Science and Humanities, Prince Sattam bin Abdulaziz University (PSAU), Alkharj, 11942, Kingdom of Saudi Arabia. alamprez@gmail.com.|Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi, 110025, India. alamprez@gmail.com.
authors.forename: Pravej
authors.initials: P
authors.lastname: Alam
authors.affiliation: Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi, 110025, India.
authors.forename: Zainul Abdeen
authors.initials: ZA
authors.lastname: Khan
authors.affiliation: Department of Biotechnology, Faculty of Science, Centre for Transgenic Plant Development, Jamia Hamdard, New Delhi, 110062, India.
authors.forename: Malik Zainul
authors.initials: MZ
authors.lastname: Abdin
authors.affiliation: Department of Biosciences, Faculty of Natural Sciences, Jamia Millia Islamia, New Delhi, 110025, India.
authors.forename: Jawaid A
authors.initials: JA
authors.lastname: Khan
authors.affiliation: Department of Botany and Microbiology, Faculty of Science, King Saud University, Riyadh, 11451, Saudi Arabia.|Department of Botany, S.P. College, Srinagar, 190001, Jammu and Kashmir, India.
authors.forename: Parvaiz
authors.initials: P
authors.lastname: Ahmad
authors.affiliation: Sara Alghonaim Research Chair (SRC), Biology Department, College of Science and Humanities, Prince Sattam bin Abdulaziz University (PSAU), Alkharj, 11942, Kingdom of Saudi Arabia.|Plant Transformation and Biopharmaceuticals Lab, Agricultural Genetic Engineering Research Institute (AGERI), Agricultural Research Centre (ARC), Giza, Egypt.
authors.forename: Shereen F
authors.initials: SF
authors.lastname: Elkholy
authors.affiliation: Sara Alghonaim Research Chair (SRC), Biology Department, College of Science and Humanities, Prince Sattam bin Abdulaziz University (PSAU), Alkharj, 11942, Kingdom of Saudi Arabia.|Department of Medicinal and Aromatic Plants Research, National Research Centre (NRC), Cairo, 12622, Egypt.
authors.forename: Mahmoud A
authors.initials: MA
authors.lastname: Sharaf-Eldin
keywords: Agrobacterium; BAP; Catharanthus roseus; GFP; Hypocotyl; Internode; NAA; Regeneration; SAAT