2023-05-11
Agrobacterium tumefaciens-mediated leaf disc genetic transformation is an important way to achieve transgenics or gene editing. Desipe of over half a century's research on related field, ensuring stable and efficient genetic transformation is still an important problem in modern biology.
Recently, the research results entitled "Effect of a suitable treatment period on the genetic transformation efficiency of the plant leaf disc method" from the research group led by the Professor Kang Xiangyang from the College of Biological Sciences and Biotechnology of BFU was published in the journal of Plant Methods(Q1). The study provides a set of simple and efficient methods to determine the optimal treatment period of genetic transformation using the leaf disc method and provides theoretical support for other species to establish an efficient and stable genetic transformation system.
It is assumed that the difference in the development status of genetic transformation cells of receptor materials is the main reason for the difference and instability of genetic transformation efficiency; the stable and efficient genetic transformation rate can be obtained by defining the appropriate treatment period of the receptor material and applying genetic transformation in a timely manner. Based on these assumptions, the research team studied and established an efficient and stable Agrobacterium-mediated plant transformation system with hybrid poplar (Populus alba × Populus glandulosa, 84 K) leaves, stem segments and tobacco leaves as the research objects. There were differences in the development process of leaf bud primordial cells from different explants, and the genetic transformation efficiency was significantly related to the cell development stage of the in vitro cultured materials. Among them, the genetic transformation rate of poplar and tobacco leaves was the highest on the 3rd and 2nd day of culture, reaching 86.6% and 57.3%, respectively. The genetic transformation rate of poplar stem segments was the highest on the 4th day of culture, reaching 77.8%. The best treatment period was from the development of leaf bud primordial cells to the S phase of the cell cycle. The number of cells detected using flow cytometry and 5-ethynyl-2ʹ-deoxyuridine (EdU) staining, the expression of cell cycle-related protein CDKB1; 2, CDKD1; 1, CYCA3; 4, CYCD1; 1, CYCD3; 2, CYCD6; 1, and CYCH; 1 of explants, and morphological changes of explants can be used as indicators to determine the appropriate treatment period for genetic transformation.
The study provides a new and universal set of methods and characteristics to identify the S phase of the cell cycle and apply genetic transformation treatments at the appropriate time, and the results are of great significance for improving the efficiency and stability of plant leaf disc genetic transformation.
The first author of the article is Xia Yufei, a doctoral candidate from the College of Biological Sciences and Biotechnology, and the coresponding author is the Professor Kang Xianyang. This research was supported by the National Key R&D Program of China during the 14th Five-year Plan Period (2021YFD2200100). Cao Yuan from the State Key Laboratory of Tree Genetics and Breeding of Chinese Academy of Forestry, doctoral candidate Ren Yongyu and Ling Aoyu, Postdoctor Du Kang, lecturer Li Yun and Yang Jun all made contributions to the research.
Paper Link: https://doi.org/10.1186/s13007-023-00994-3
