Molecular Characterization of Granule-Bound Starch Synthase (GBSSI) gene of Waxy Locus Mutants in Japonica Rice (Oryza sativa L.)

  • Sohn, Seong-Han (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Rhee, Yong (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Hwang, Duk-Ju (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Lee, Sok-Young (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Lee, Jung-Ro (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Lee, Yeon-Hee (National Academy of Agricultural Science (NAAS), Rural Development Administration (RDA)) ;
  • Shin, Young-Seop (National Institute of Crop Science (NICS), Rural Development Administration (RDA)) ;
  • Jeung, Ji-Ung (National Institute of Crop Science (NICS), Rural Development Administration (RDA)) ;
  • Kim, Myung-Ki (National Institute of Crop Science (NICS), Rural Development Administration (RDA))
  • 투고 : 2010.03.10
  • 발행 : 20100300

초록

Five mutants were investigated at the molecular level to determine the factors responsible for mutated endosperm types. They were classified as high (HA) or low amylose (LA) phenotypes based on the amylose content in endosperm. The five were previously produced from Ilpum and Shindongjin cultivar treated with N-methyl-N-nitrosourea and gamma-ray irradiation, respectively. Analysis of the genomic structure and expression of Granule-bounded Starch Synthase I (GBSSI) genes revealed that mutants generally showed a higher incidence of nucleotide transition than transversion, and the $A:T{\rightarrow}G:C$ transition was particularly prevalent. The rates of nucleotide substitution in HA mutants were generally higher than those in the LA mutants, leading to higher substitutions of amino acid in the HA mutants. Neither nucleotide substitutions interfering with intron splicing or causing early termination of protein translation were found, nor any large-sized deletions or additions were found in all the mutants. In principle, amylose content can be regulated by three factors: internal alterations of GBSSI protein, the strength of gene expression, and other unknown external factors. Our results indicate that the endosperm mutants from Shindongjin arose from internal alterations of GBSSI proteins, which may be the result of amino acid substitutions. On the other hand, the Ilpum mutants might be principally caused by the alteration of gene expression level. Analysis of another three glutinous cultivars revealed that the major factor leading to glutinous phenotypes is the 23-bp duplicative motif (5'-ACGGGTTCCAGGGCCTCAAGCCC-3') commonly found in exon 2, which results in the premature termination of protein translation leading to the production of a non-functional GBSSI enzyme.

키워드

과제정보

연구 과제 주관 기관 : Rural Development Administration(RDA)

참고문헌

  1. Baldwin P. 2001. Starch granule-associated proteins and polypeptides. Starch 53:475-503. https://doi.org/10.1002/1521-379X(200110)53:10<475::AID-STAR475>3.0.CO;2-E
  2. Denyer K. Johnson P. Zeeman S. Smith AM. 2001. The control of amylose synthesis. J Plant Physiol 158:479-487. https://doi.org/10.1078/0176-1617-00360
  3. Cai XL, Wang ZV, Xing YY, Zhang JL, Hong MM. 1998. Aberrant splicing of intron 1 leads 10 the heterogeneous 5' UTR and decreased expression of waxy, gene in rice cultivars of intermediate amylose content. Plant J 14:459-465. https://doi.org/10.1046/j.1365-313X.1998.00126.x
  4. Chin JH, Kim JH, Jiang W, Chu SH, Woo MO, Han L, Brar D, Koh HJ. 2007. Identification of Subspecies-specific STS Markers and Their Association with Segregation Distortion in Rice (Oryza saliva L.). J Crop Sci Biotech 10:175-184.
  5. Colbert T, Till BJ, Tompa R, Reynolds S, Steine MN, Yeung AT, MeCallum CM, Comai L, McCallum CM, Comai L. Greene EA, Henikoff S. 2000. Targeted screening for induced mutations. Nat Biotech 18:455-457. https://doi.org/10.1038/74542
  6. Erog lu A, Ulu A, Kurtman C, Cam R, Akar N. 2006. 23-bp endothelial protein C receptor (eper) gene insertion mutation in cancer patients with and without thrombosis, Am J Hematol 81:220.
  7. Henikoff S. 2001. High-Throughput Screening for Induced Point Mutations. Plant Physiol 126:480-484. https://doi.org/10.1104/pp.126.2.480
  8. Hirano HY, Sano Y, 1991. Molecular Characterization of the waxy Locus of Rice (Oryza saliva). Plant Cell Physiol 32:989-997.
  9. Hori Y, Fujimoto R, Sato Y, Nishio T, 2007. A novel wx mutation caused by insertion of a retrotransposon-like sequence in a glutinous cultivar of rice (Oryza saliva). Theor App Genet 115:217-224. https://doi.org/10.1007/s00122-007-0557-6
  10. Inukai T, Sako A, Hirano HY, Sano Y. 2000. Analysis of intragenic recombination at wx in rice: correlation between the molecular and genetic maps within the locus. Genome 43:589-96.
  11. Isshiki M, Morino K, Nakajima M, Okagaki RJ, Wessler SR, Izawa T, Shinmmoto K, 1998. A naturally occurring functional allele of the rice waxy locus has a GT to TT mutation at thc 5' splice site of the first intron. Plant J 15:133-138. https://doi.org/10.1046/j.1365-313X.1998.00189.x
  12. Kaare M, Ulander VM, Painter JN, Ahvenainen T, Kaaja R, Aittomaki K. 2007. Variations in the thrombomodulin and endothelial protein C receptor genes in couples with recurrent miscarriage. Human Reproduct 22:864-868.
  13. MeCallum CM, Comai L, Greene EA, Henikoff S. 2000. Targeting Induced Local Lesions IN Genomes (TILLING) for Plant Functional Genomics. Plant Physiol 123: 439-442. https://doi.org/10.1104/pp.123.2.439
  14. Olsen O, Wang X, von Wettstein D. 1993. Sodium azide mutagenesis: Preferential generation of $A.T\{rightarrow}G.C$ transitions in the barley Ant18 gene. Proc Natl Acad Sci USA: 043-8047.
  15. Perez CM, Juliano BO. 1978. Modification of the simplified amylose test for milled rice. Starch 30: 424-426. https://doi.org/10.1002/star.19780301206
  16. Shapter FM, Eggler P, Lee LS, Henry RJ. 2009. Variation in Granule Bound Starch Synthase I (GBSSI) loci amongst Australian wild cereal relatives (Poaceae). J Cereal Sci 49:4-11. https://doi.org/10.1016/j.jcs.2008.06.013
  17. Taira T, Fujita N, Takaoka K, Ucmatsu M, Wadano A, Kozaki S, Okabe S. 1995. Variation in the primary structure of waxy proteins (granule-bound starch synthase) in diploid cereals. Biochem Genet 33:269-281.
  18. Umeda M, Ohtsubo H, Ohtsubo E. 1991. Diversification of the rice Waxy gene by insertion of mobile DNA elements into introns. Jpn J Genet 66:569-586. https://doi.org/10.1266/jjg.66.569
  19. Wanchana S, Toojinda T, Tragoonrung S, Vanavichil A. 2003. Duplicated coding sequence in the waxy allele of tropical glutinous rice (Oryza saliva L.). Plant Sci 165:1193-1199. https://doi.org/10.1016/S0168-9452(03)00326-1
  20. Wang ZY, Zheng FQ, Shen GZ, Gao JP, Snustad P, Li MG, Zhang JL, Hong MM. 1995. The amylose content in rice endosperm is related to the post-transcriptional regulation of the waxy gene. Plant J 7:613-622. https://doi.org/10.1046/j.1365-313X.1995.7040613.x