RESEARCH ARTICLE

Construction of yeast two-hybrid cDNA libraries for wheat near-isogenic line TcLr19 under the stress of Puccinia recondita and its preliminary appreciation

  • Lifeng ZHANG ,
  • Hui ZHOU ,
  • Fengju WEI ,
  • Ziyi CHENG ,
  • Aihua YAN ,
  • Dongmei WANG
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  • College of Life Science, Agricultural University of Hebei, Baoding 071001, China

Received date: 09 Nov 2010

Accepted date: 06 May 2011

Published date: 05 Dec 2011

Copyright

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg

Abstract

Two cDNA libraries for wheat near-isogenic line TcLr19 under Puccinia recondita stress were constructed by using SMART technique and homologous reorganization method. Wheat near-isogenic line TcLr19 was infected with leaf rust race 366, and total RNA was extracted from the leaves after infection for 4, 8, and 12 h. The total RNAs were reverse transcribed to cDNA by using oligo(dT) primer and random primer, respectively. According to the evaluation on quality, the transformation efficiency was about 1.32 × 106 and 1.0 × 106 transformants/3 μg pGADT7-Rec, respectively, and the library titers were up to 2.62 × 108 and 3.51 × 108 pfu/mL, with 93% and 100% recombinant rate, which indicated the high quality of the two libraries for next screening.

Cite this article

Lifeng ZHANG , Hui ZHOU , Fengju WEI , Ziyi CHENG , Aihua YAN , Dongmei WANG . Construction of yeast two-hybrid cDNA libraries for wheat near-isogenic line TcLr19 under the stress of Puccinia recondita and its preliminary appreciation[J]. Frontiers of Agriculture in China, 0 , 5(4) : 450 -455 . DOI: 10.1007/s11703-011-1123-1

Acknowledgements

This research was supported by the National Natural Science Foundation of China (Grant Nos. 30671244 and 31171472), the Key Basic Research Project of Applied Basic Research Program of Hebei Province, China (No. 08965505D), and the Natural Science Foundation of Hebei Province, China (Nos. C2007000515 and C2010000787).
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