Agrobacterium tumefaciens-mediated transformation of a hevein-like gene into asparagus leads to stem wilt resistance


Autoři: Helong Chen aff001;  Anping Guo aff001;  Zhiwei Lu aff004;  Shibei Tan aff003;  Jian Wang aff001;  Jianming Gao aff002;  Shiqing Zhang aff002;  Xing Huang aff003;  Jinlong Zheng aff003;  Jingen Xi aff003;  Kexian Yi aff003
Působiště autorů: Institute of Tropical Agriculture and Forestry, Hainan University, Haikou, China aff001;  Institute of Tropical Bioscience and Biotechnology/Hainan Academy of Tropical Agricultural Resource, Chinese Academy of Tropical Agricultural Sciences /Key Laboratory of Biology and Genetic Resources of Tropical Crops, Ministry of Agriculture, Haikou, Chi aff002;  Environment and Plant Protection Institute, Chinese Academy of Tropical Agricultural Sciences, Haikou, Hainan, China aff003;  South Subtropical Crops Institute, Chinese Academy of Tropical Agricultural Sciences/ Zhanjiang City Key Laboratory for Tropical Crops Genetic Improvement, Zhanjiang, Guangdong, China aff004
Vyšlo v časopise: PLoS ONE 14(10)
Kategorie: Research Article
doi: 10.1371/journal.pone.0223331

Souhrn

Asparagus stem wilt, is a significant and devastating disease, typically leading to extensive economic losses in the asparagus industry. To obtain transgenic plants resistant to stem wilt, the hevein-like gene, providing broad spectrum bacterial resistance was inserted into the asparagus genome through Agrobacterium tumefaciens-mediated transformation. The optimal genetic transformation system for asparagus was as follows: pre-culture of embryos for 2 days, inoculation using a bacterial titre of OD600 = 0.6, infection time 10 min and co-culturing for 4 days using an Acetosyringone concentration of 200 μmol/L. Highest transformation frequencies reached 21% and ten transgenic asparagus seedlings carrying the hevein-like gene were identified by polymerase chain reaction. Moreover, integration of the hevein-like gene in the T1 generation of transgenic plants was confirmed by southern blot hybridization. Analysis showed that resistance to stem wilt was enhanced significantly in the transgenic plants, in comparison to non- transgenic plants. The results provide additional data for genetic improvement and are of importance for the development of new disease-resistant asparagus varieties.

Klíčová slova:

Agrobacteria – Embryos – Genetically modified plants – Plant pathology – Plasmid construction – Polymerase chain reaction – Seedlings – Agrobacterium tumefaciens


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