African Journal of
Biotechnology

  • Abbreviation: Afr. J. Biotechnol.
  • Language: English
  • ISSN: 1684-5315
  • DOI: 10.5897/AJB
  • Start Year: 2002
  • Published Articles: 12487

Full Length Research Paper

Full-length enriched multistage cDNA library construction covering floral bud development in Populus tomentosa

  Xin-Min An, Dong-Mei Wang, Ze-Liang Wang, Mei-Xia Ye and Zhi-Yi Zhang*  
National Engineering Laboratory for Tree Breeding; Key Laboratory of Genetics and Breeding in Forest Trees and Ornamental Plants, Ministry of Education; Tree and Ornamental Plant Breeding and Biotechnology Laboratory of State Forestry Administration, Beijing Forestry University, Beijing 100083, P. R. China.
Email: [email protected]

  •  Accepted: 12 August 2011
  •  Published: 10 April 2012

Abstract

 

Flowering involves expression of a suite of genes associated with floral development. The genome of the Chinese white poplar (Populus trichocarpa) was sequenced because of its importance as a model tree for genetic studies as well as being an economically important woody plant. However, information on expressed genes involved in poplar floral bud development is insufficient to allow annotation of genes and use of the genomic information. To isolate and characterize genes involved in flowering of Populus tomentosa, floral bud samples were collected at different developmental stages from floral bud initiation to flower maturity, and full-length enriched cDNA libraries from both male and female floral buds were constructed. The results of titer analysis showed that the titer of the female and male primary libraries were 8.00 × 10and 7.20 × 10pfu/ml, respectively, and the titer of the amplified libraries were 2.60 × 10and 2.56 × 10pfu/ml, respectively. The combination ratio reached 90% and the insert size was 400 to 2000 bp. The results indicated that cDNA libraries were successfully constructed.

 

Key words: cDNA library, floral bud, flowering, Populus tomentosa.

Abbreviation

Abbreviations: DMSO, Dimethyl sulfoxide; LD-PCR, long-distance polymerase chain reaction; dscDNA, double-strand complementary DNA; pfu, plaque-forming unit.