[The analysis of differential expression and cloning of genes related to raised secondary lateral veins mutant of Lycoris aurea].
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 microarray 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.
Gao
Lei
L
The Key Laboratory of Forest Genetics and Gene Engineering, Nanjing Forestry University, Nanjing 210037, China. lgao528@yahoo.com.cn
Guo
Su-Min
SM
Cui
Yong-Lan
YL
Zhuge
Qiang
Q
Huang
Min-Ren
MR
D000595:Amino Acid Sequence; D019411:Clinical Laboratory Techniques; D019976:Cloning, Organism; D018076:DNA, Complementary / Q000302:isolation & purification*; D015870:Gene Expression; D020869:Gene Expression Profiling; D017343:Genes, Plant / Q000235:genetics* / Q000502:physiology; D031426:Lycoris / Q000235:genetics* / Q000254:growth & development*; D008969:Molecular Sequence Data; D020411:Oligonucleotide Array Sequence Analysis; D018515:Plant Leaves / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010946:Plants, Medicinal / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment
D018076:DNA, Complementary