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HEREDITAS ›› 2012, Vol. 34 ›› Issue (4): 472-484.doi: 10.3724/SP.J.1005.2012.00472

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Cloning and expression analysis of differentially expressed genes in Chinese fir stems treated by different concentrations of exogenous IAA

YANG Li-Wei, SHI Ji-Sen   

  1. Key Lab of Forest Genetics and Biotechnology of the Ministry of Education of China, College of Forest Resources and Environment, Nanjing Forestry University, Nanjing 210037, China
  • Received:2011-09-05 Revised:2012-02-09 Online:2012-04-20 Published:2012-04-25

Abstract: To reveal the potential genetic mechanisms of indole-3-acetic acid (IAA) that regulate Chinese fir wood formation, cloned the differentially expressed genes via suppress subtractive hybridization (SSH) using the truncated stems treated by 0 and 3 mg IAA/g lanolin as the driver and tester, respectively. A total of 332 unigenes that were involved in cell organization and biosynthesis, developmental processes control, electron transport, stress response, and signal transduction. To further test the results from SSH, we selected those unigenes, whose putative encoding proteins showed significantly homologous with HIRA, PGY1, SMP1, TCT, TRN2, and ARF4, and analyzed their expressed specificity in the wood formative tissues and their response to the secondary developmental changes of vascular cambium stimulated by 0, 1, and 3 mg.IAA/g.lanolin treatment. The results showed that ClHIRA, ClPGY1, and ClARF4, which were specifically expressed in the adaxial zone of stem, were positively response to the activities of cell division and tracheid differentiation stimulated by exogenous IAA treatment. However, ClSMP1, ClTCTP1, and ClTRN2, which were mainly expressed in the abaxial zones of stems, showed negative correlation with the treated levels of exogenous IAA and activities of vascular cambium secondary development at the transcriptional level. This result showed that the differential response of develop-mental regulatory genes located in different vascular tissues to the level changes of edogenous IAA in stems is likely to be an important molecular mechanism of auxin regulating wood formation.

Key words: exogenous IAA, Cunninghamia lanceolata (Lamb.) Hook, stems, the cloning of differentially ex-pressed genes, expression analysis