Expression of insect (Microdera puntipennis dzungarica) antifreeze protein MpAFP149 confers the cold tolerance to transgenic tobacco.
To elucidate the function of antifreeze protein from Microdera puntipennis dzhungarica for freezing stress tolerance in plant, the construct of MpAFP149 gene with the signal peptide sequence responsible for secreting the native MpAFP149 into the apoplast space under control of a cauliflower mosaic virus 35S promoter was introduced into tobacco by Agrobacterium tumefaciens-mediated transformation. The observation of immunogold localization by TEM (transmission electron microscope) showed that the heterologous MpAFP149 protein was mainly distributed on the cell wall in apoplast of the transgenic tobacco plant. T1 generation transgenic tobacco plants displayed a more frost resistant phenotype and kept the lower ion leakage ratio and MDA (malondialdehyde) content in the leaves compared with wild-type ones at -1 degrees C for 3 days. The results showed that MpAFP149 provided protection and conferred cold tolerance to transgenic tobacco plant during freezing stress.
Wang
Yan
Y
Xinjiang Key Laboratory of Biological Resources and Genetic Engineering, College of Life Science and Technology, Xinjiang University, 14 Shengli Road, 830046, Urumqi, China.
Qiu
Liming
L
Dai
Chunying
C
Wang
Jing
J
Luo
Jianmin
J
Zhang
Fuchun
F
Ma
Ji
J
D000222:Adaptation, Physiological / Q000235:genetics / Q000502:physiology; D000818:Animals; D021301:Antifreeze Proteins / Q000235:genetics* / Q000502:physiology; D017796:Caulimovirus / Q000235:genetics; D001517:Coleoptera / Q000235:genetics / Q000378:metabolism; D005615:Freezing; D018506:Gene Expression Regulation, Plant; D019476:Insect Proteins / Q000235:genetics / Q000502:physiology; D046529:Microscopy, Electron, Transmission; D030821:Plants, Genetically Modified / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure; D011401:Promoter Regions, Genetic / Q000235:genetics; D014026:Nicotiana / Q000235:genetics* / Q000502:physiology / Q000648:ultrastructure
D021301:Antifreeze Proteins; D019476:Insect Proteins