Rizwan Hafiz Muhammad, Yang Qiang, Yousef Ahmed Fathy, Zhang Xiaoxue, Sharif Yasir, Kaijie Jia, Shi Meng, Li Han, Munir Nigarish, Yang Xuelian, Wei Xiaoxia, Oelmüller Ralf, Cheng Chunzhen, Chen Faxing
College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, China.
Department of Horticulture, College of Agriculture, University of Al-Azhar (Branch Assiut), Assiut 71524, Egypt.
Plants (Basel). 2021 Nov 15;10(11):2459. doi: 10.3390/plants10112459.
Passion fruit () is an important fruit crop with high economic value. Genetic engineering plays an important role in crop improvement with desired traits and gene functional studies. The lack of a simple, efficient, and stable transformation system for passion fruit has greatly limited gene functional studies. In this study, a simple and efficient -mediated transformation system for passion fruit was established, using virulent strain EHA105 harboring the binary vectors pCAMBIA1301 and pCAMBIA1302 with GUS and GFP reporter genes. The system requires less time and labor costs than conventional transformation systems, and no additional phytohormones and sterile conditions are required. Regeneration efficiency of 86% and transformation efficiency of 29% were achieved, when the wounds were wrapped with Parafilm and the plants were kept in darkness for 15 days. Approximately 75% of the regenerated plants had a single shoot and 26% multiple shoots. The transformation was confirmed at the DNA and RNA levels as well as by GUS staining and GFP fluorescent measurements. The developed protocol will contribute to the genetic improvement of passion fruit breeding.
西番莲是一种具有高经济价值的重要水果作物。基因工程在培育具有理想性状的作物和基因功能研究中发挥着重要作用。西番莲缺乏简单、高效且稳定的转化系统，这极大地限制了基因功能研究。在本研究中，利用携带含有GUS和GFP报告基因的双元载体pCAMBIA1301和pCAMBIA1302的强毒株EHA105，建立了一种简单高效的西番莲农杆菌介导的转化系统。该系统比传统转化系统所需时间和劳动力成本更少，且无需额外的植物激素和无菌条件。当用封口膜包裹伤口并将植株置于黑暗中15天时，再生效率达到86%，转化效率达到29%。大约75%的再生植株有单芽，26%有多芽。通过DNA和RNA水平以及GUS染色和GFP荧光测量对转化进行了确认。所建立的方案将有助于西番莲育种的遗传改良。
