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HEREDITAS ›› 2007, Vol. 29 ›› Issue (4): 490-490―498.doi: 10.1360/yc-007-0490

• 研究报告 • Previous Articles     Next Articles

The analysis of differential expression and cloning of genes related to raised secondary lateral veins mutant of Lycoris aurea

GAO Lei1,2,3, GUO Su-Min1, CUI Yong-Lan1,4, ZHUGE Qiang1, HUANG Min-Ren1

  

  1. 1. The Key Laboratory of Forest Genetics and Gene Engineering, Nanjing Forestry University, Nanjing 210037, China;
    2. Wuhan Botanical Garden, Chinese Academy of Sciences, Wuhan 430074, China;
    3. Graduate School of Chinese Academy of Sciences, Beijing 100049, China;
    4. College of Life of Shanghai Normal University, Shanghai 200234, China
  • Received:2006-07-21 Revised:2006-10-09 Online:2007-04-10 Published:2007-04-10
  • Contact: HUANG Min-Ren

Abstract:

Lycoris aurea exhibits parallel venation, the main vein with many lateral veins in a longitudinal parallel arrangement. There are secondary lateral veins (SLV) between each longitudinal veins. In general, SLVs are not remarkable. In this paper, the material was one kind of Lycoris aurea mutant called Raised Secondary Lateral Veins mutant (RSLV), because many Raised Secondary Lateral Veins are in abaxial surface of its leaves. Its growing potential is weaker than that of wild type and its blades are very thin. Moreover, the stamens of RSLV degenerate completely. Two cDNA libraries were constructed from RSLV mutant and wild type (WT) leaves. From the libraries, 3,122 ESTs, which are longer than 100 bp each after vector sequence removed, were acquired by single-pass sequencing from the 5′end. Following a multistep

selection, 512 70-mer oligo-DNA probes were designed for attachment on the microarray slide based on the ESTs. The gene expression profile of RSLV mutant and WT leaves was compared through the microarray at transcriptional level. The mi-croarray experiment results were further confirmed by Quantitative Real-Time PCR (QRT-PCR). We identified 5 genes whose expressions changed more than 2-fold between RSLV mutant and WT leaves. They encode phloem protein 2 (PP2), ferritin, pectin methyl esterase (PME), chlorophyll a/b binding protein (CAB protein) and pyruvate decarboxylase (PDC), respectively. Furthermore, the full-length cDNA sequences of the 5 genes were separately obtained from RSLV and WT by RACE. The relationship between differential expressions of the genes and the formation of the RSLV mutant phenotype were discussed.