Punctual mutations in lipoxygenase sequence expressed in black soybean seed coat

Mutações pontuais em sequências expressas da enzima lipoxigenase em tegumentos de sementes de soja de coloração preta


Lipoxygenase enzymes are relevant in several characteristics of soybean seeds, such as germination performance, storage capacity and tolerance to infection by microorganisms. In this sense, the study of allelic variations of the genes related to that enzyme may be promising for the development of molecular markers associated with phenotypic characteristics which are relevant to seed physiology. This study aimed to identify mutations in lipoxygenase sequence expressed in black soybean seed coat. After isolation of the sequence of interest by the cDNA-AFLP technique, such sequence was compared with sequences of nucleotides and proteins of the lipoxygenase present in yellow soybean seed coat. Analysis of multiple sequence alignment has shown an expressive number of changes in the sequence of nucleotides, which have resulted in the modification of 13 amino acids in the translated protein. The identification of such mutations widens the possibility for further studies involving the association of these allelic variations with phenotypic differences of interest to seeds physiology.

Glycine max; molecular marker; enzymes

Enzimas lipoxigenases possuem relevância em diversas características das sementes de soja, como desempenho germinativo, capacidade de armazenamento e tolerância ao ataque de microrganismos. Nesse sentido, o estudo de variações alélicas de genes relacionados a essa enzima podem ser promissores para o desenvolvimento de marcadores moleculares relacionados a caracteres fenotípicos de interesse a fisiologia de sementes. Este trabalho objetivou identificar mutações em sequência de lipoxigenase expressa em tegumentos de sementes de soja de coloração preta. Após isolamento da sequência de interesse pela técnica de cDNA-AFLP, a mesma foi comparada com sequências de nucleotídeos e proteínas da lipoxigenase presente em sementes de soja de coloração amarela. A análise do alinhamento múltiplo entre as sequências mostrou um número expressivo de alterações na sequência de nucleotídeos, as quais resultaram na modificação de 13 aminoácidos na proteína traduzida. A identificação dessas mutações abre a possibilidade para estudos futuros envolvendo a associação dessas variações alélicas com diferenças fenotípicas para caracteres de interesse a fisiologia de sementes.

Glycine max; marcadores moleculares; enzimas

  • AKRAM, A.; ONGENA, M.; DUBY, F.; DOMMES, J.; THONART, P. Systemic resistance and lipoxygenase-related defence response induced in tomato by Pseudomonas putida strain BTP1. BMC Plant Biology, v.8, n.113, 2008. http://www.biomedcentral.com/1471-2229/8/113
  • AXELROD, V.; CHEESBROUGH, T.M.; LAASKO, S. Lipoxygenase from soybeans. Methods in Enzymology, v.71, p.441-451, 1981.
  • DIAS, A.C.P.; REIS, M.S.; OLIVEIRA, M.G.A.; SEDIYAMA, C.S.; MOREIRA, M.A. Atividade de lipoxigenases durante a germinação de sementes de soja. Revista Ceres, v.49, n.284, p.419-30, 2002. http://www.ceres.ufv.br/ceres/revistas/v49n284p16602.pdf
  • GAO, X.; BRODHAGEN, M.; ISAKEIT, T.; BROWN, S.H.; GOBEL, C.; BETRAN, J.; FEUSSNER, I., KELLER, N.P.; KOLOMIETS, M.V. Inactivation of the lipoxygenase ZmLOX3 increases susceptibility of maize to Aspergillus spp. In: Molecular Plant Microbe Interactions, v.22, p.222-231, 2009.
  • GASTEIGER, E.; GATTIKER, A.; HOOGLAND, C.; IVANYI, I.; APPEL, R.D.; BAIROCH, A. ExPASy: the proteomics server for in-depth protein knowledge and analysis. Nucleic Acids Research, v.31, p.3784-3788, 2003.
  • HALL, T.A. BioEdit: a user-friendly biological sequence alignment editor and analysis program for Windows 95/98/NT. Nucleic Acids Symposium, 1999. 41p.
  • LENIS, J.M.; GILLMAN, J.D.; LEE, J.D.; SHANNON, J.G.; BILYEU, K.D. Soybean seed lipoxygenase genes: molecular characterization and development of molecular marker assays. Theorical and Applied Genetics, v.120, p.1139-1149, 2010. http://link.springer.com/article/10.1007%2fs00122-009-1241-9
  • LIMA, W.A.A.; BORÉM, A.; DIAS, D.C.F.S.; MOREIRA, M.A.; DIAS, L.A.S. Lipoxygenase and physiological quality of soybean seeds during storage. Seed Science and Technology, v.38, n.3, p.767-771, 2010.
  • MALONE, G.; PESKE, S.T.; ZIMMER, P.D.; MALONE, E.; MENEGHELLO, G.E.; OLIVEIRA, C.A. Single nucleotide polymorphism (SNP) detection in the red rice α-amylase gene amy1: effect on seedling vigor. Seed Science and Technology, v.36, n.2, p.447-445, 2008.
  • MERTZ, L.M.; HENNING, F.A.; BORSUK, S.; MAIA, L.C.; DELLAGOSTIN, O.A.; PESKE, S.T.; ZIMMER, P.D. cDNA-AFLP analyses between black and yellow soybean seed coats. Seed Science and Technology, v.38, p.88-95, 2010.
  • MO, B.; KOSTER, K.L. Changes in lipoxygenase isoforms during germination and early seedling growth of Pisum sativum L. Seed Science Research, v.16, p.97-106, 2006.
  • NCBI. National Center for Biotechnology Information http://www.ncbi.nlm.nih.gov. Accessed on: Jan. 13rd 2012.
  • OLIVEIRA, D.A.; PIOVESAN, N.D.; JOSÉ, I.C.; BARROS, E.G.; DIAS, D.C.F.S.; MOREIRA, M.A. Lipoxigenases e teor de ácido linolênico relacionados à qualidade de sementes de soja. Revista Brasileira de Sementes, v.28, n.1, p.30-35, 2006. http://www.scielo.br/pdf/rbs/v28n1/a05v28n1.pdf
  • SIEDOW, J.N. Plant lipoxygenase: structure and function. Annual Reviews Plant Physiology Plant Molecular Biology, v.42, p.145 - 188, 1991. http://www.annualreviews.org/doi/pdf/10.1146/annurev.pp.42.060191.001045
  • SKRZYPCZAK-JANKUN, E.; AMZEL, L.M.; KROA, B.A.; JR. FUNK, M.O. Proteins: Structure, Function and Genetic, v.29, p.15-31, 1997.
  • THOMPSON, J.D.; HIGGINS, D.G.; GIBSON, T.J. CLUSTAL W. Improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Research, v.22, n.22, p.4673-4680, 1994. http://www.ncbi.nlm.nih.gov/pmc/articles/pmc308517/pdf/nar00046-0131.pdf
  • VICK, B.A.; ZIMMERMAN, D.C. Lipoxygenase, hydroperoxide isomerase and hydroperoxide cyclase in young cotton seedlings. Plant Physiology, v.67, p.92-97, 1981. http://www.plantphysiol.org/content/67/1/92.full.pdf
  • WANG, J.Y.; FUJIMOTO, K.; MIYAZAWA, T.; ENDO, Y.; KITAMURA, K. Sensitivities of lipoxygenase-lacking soybean seeds to accelerated aging and their chemiluminescence levels. Phytochemistry, v.29, n.12, p.3739-3742, 1990. http://www.sciencedirect.com/science/article/pii/0031942290853238
  • WANG, C.; CROFT, K.P.C.; JARLFORS, U.; HILDEBRAND, D.F. Subcellular localization studies indicate that lipoxygenases 1 to 6 are not involved in lipid mobilization during soybean germination. Plant Physiology, v.120, p.227 - 236, 1999. http://www.ncbi.nlm.nih.gov/pubmed/10318700

Publication Dates

  • Publication in this collection
    21 June 2013
  • Date of issue


  • Received
    25 Oct 2012
  • Accepted
    13 Feb 2013
ABRATES - Associação Brasileira de Tecnologia de Sementes Avenida Maringá, nº 1219 , Jardim Vitória Londrina - Paraná Brasil, Tel./ Fax. 55 43 3025-5120 - Londrina - PR - Brazil
E-mail: contato@abrates.org.br