[Initial functional analysis of the promoter region and coding region of Pib gene in transgenic rice].
The promoter region and intact coding region of Pib gene (9.9 kb) was inserted into the downstream of CaMV 35S promoter in a binary vector pPZP2Ha3(+), resulting a plasmid pNAR701. And a fragment of Pib gene from 6 986 to 9 392 bp was placed into pPZP2Ha3(-) under the control of CaMV 35S promoter, producing an antisense expression vector pNAR703. These two recombined vectors were transferred into a blast medium susceptible rice cultivar R109 by an Agrobecterium-mediated method. Tests of PCR and Southern blotting for transgenic plants as well as the segregation of hygromycin resistance in T1 generation confirmed that the target DNA fragments were integrated into genome of R109 and hereditable. Northern blotting analysis showed the coding region of Pib gene double driven by 35S and its native promoter was able to transcript in T1 transgenic plants. Rice blast resistance test for T1 transgenic seedlings of 3-4 leaves stage and in vitro leaves in tillering stage showed that transgenic plants of pNAR701 were more resistant to blast race ZD1 and ZG1 than the wild type plants, but the resistance of antisense transgenic plants from pNAR703 was decreased compared to the controls.
Zhou
Ming
M
National Key Laboratory of Crop Genetics and Germplasm Enhancement, Nanjing Agricultural University, Nanjing 210095, China.
Yang
Shi-Hu
SH
Lan
Ying
Y
Jin
Yu-Kuan
YK
Wan
Jian-Min
JM
D015139:Blotting, Southern; D017343:Genes, Plant / Q000235:genetics* / Q000502:physiology; D005822:Genetic Vectors / Q000235:genetics; D008957:Models, Genetic; D012275:Oryza / Q000235:genetics*; D030821:Plants, Genetically Modified / Q000235:genetics*; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000235:genetics*