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Cloning and Expression Analysis of AP3-like Gene in Oncidium

Author: XuXiaoYan
Tutor: LongMingHua；TianMin
School: Guangxi University
Course: Crop Cultivation and Farming System
Keywords: Floral organ formation MADS-box Oncidium APETALA3 OAP3 gene qRT-PCR
CLC: S682.31
Type: Master's thesis
Year: 2011
Downloads: 24
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Abstract


Angiosperm AP3 family belong to the MADS-box genes in B functional genes , and its important role in the formation of floral organs and reproductive and developmental . In order to study the relationship between the orchid MADS-box gene expression and floral organ development Oncidium (Oncidium Milliongolds) , as the material this study by RT-PCR method was cloned into a flower development-related MADS-box genes , by the gene expression patterns in the flower buds at different developmental stages and different organizations floral organ formation , suggesting that the gene regulation played the role of lay the foundation for further study of the molecular mechanism for Lanhuahua organ development , provide a theoretical basis for the orchid molecular breeding . The results of this study are as follows: (1) isolated by RT-PCR , the full length of the OAP3 cDNA from the gene is 799bp , including a 612bp the CDS box , a length coding for a protein of 203 amino acids . In GenBank ( accession number : GU644447 ) . Phylogenetic analysis showed that OAP3 of floral organ development class B gene , but the gene does not exist Class B gene C-terminal domain common . (2 ) the Oncidium OAP3 gene protein belongs AP3-like gene family paleoAP3 series , compared with other species , the amino acid sequences of up to 78% of the sequence of Phalaenopsis PeMADS5 amino acid sequence homology with monocot plant rice the MADS16 and lily LMADS1 of amino acid sequence homology of 62% , significantly higher than the dicotyledon Arabidopsis AP3 (50%) and the snapdragon DEF (50%) homology . (3) OAP3 gene encoding the protein is speculated that its molecular weight is 23661.1D theoretical isoelectric point is 9.49 . Wherein the basic amino acid and total amino acid 18.23% total 13.80% of the amino acids , acidic amino acids , hydrophobic amino acids 33.00% of the total amino acid , 34.97% of the total amino acids of the polar amino acids . (4) qRT-PCR expression analysis : from start pumping flowers , with the growth of the flower buds , the gene relative expression abundance showing a rising trend ; OAP3 in sepals , petals, lips, stamen, ovary , and vegetative organs of leaves and stems in all expression .

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Perennial Flowers > Other Flowers > Orchids
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