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KMID : 0613820040140040650
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Journal of Life Science
2004 Volume.14 No. 4 p.650 ~ p.656
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Genes of Wild Rice (Oryza grandiglumis) Induced by Wounding and Yeast Extract
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Shin Sang-Hyun
Im Hyun-Hee
Lee Jai-Heon
Kim Do-Hoon
Chung Won-Bok
Kang Kyung-Ho
Chung Young-Soo
Cho Sung-Ki
Shin Jeong-Sheop
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Abstract
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Oryza grandiglumis (CCDD, 2n=48), one of the wild rice species, has been known to possess fungal-, bacterial-, and insect-resistance against sheath blight, rice blast, bacterial leaf blight and brown plant hopper (Nilaparvata lugens). To rapidly isolate differentially expressed genes responding to fungal and wounding stress, wounding and yeast extract were treated to O. grandiglumis for 24 hrs. Suppression subtractive hybridization (SSH) method was used to obtain differentially expressed genes from yeast extract and wounding treated plants. Seven hundreds and seventy six clones were obtained by subcloning PCR product, and colony array and screening were carried out using radio-isotope labeled cDNA probes prepared from the wounding and yeast extract treated plants. One hundred and fifteen colonies were confirmed as true positive ones. Average insert size of the clones were ranged from 400 bp to 700 bp and all the inserts were sequenced. To decide the identity of those clones, sequences were analyzed by sequence homology via GenBank database. The homology search result showed that 68 clones were matched to the genes with known function; 16 were related to primary metabolism, 5 to plant retrotransposons, 5 to defense related metallothionein-like genes. In addition to that, others were matched to various genes with known function in amino acid synthesis and processing, membrane transport, and signal transduction, so on. In northern blot analysis, induced expressions of ogwfi-161, ogwfi-646, ogwfi-663, and ogwfi-695 by wounding and yeast extract treatments were confirmed. The result indicates that SSH method is very efficient for rapid screening of differentially expressed genes.
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KEYWORD
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Oryza grandiglumis, SSH, yeast extract, wounding, colony array
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