lpa1-5525: A New lpa1 Mutant Isolated in a Mutagenized Population by a Novel Non-Disrupting Screening Method
Abstract
:1. Introduction
2. Results
2.1. Transposon Tagging Mutagenesis
2.2. Density Assay Screening of the Mutagenized Population
2.3. Confirmation of the HIP Phenotype of the Low Density Seeds
2.4. Molecular Analysis of the F1 Plants of the Mutagenized Population
2.5. Screening and Selection of the Putative New lpa1 Mutant
2.6. Quantitative Analysis of Phosphorus Content
2.7. Structure Analysis of the ZmMRP4 Locus
3. Discussion
4. Materials and Methods
4.1. Genetic Stocks and Sampling Material
4.2. Density Assay
4.3. Assay for Free Phosphate Content in the Seeds
4.4. Assay for Seed Phytate Content
4.5. lpa1 Locus Molecular Genotyping
4.6. Structural Analysis of ZmMRP4 Locus in Putative New lpa 1 Mutant
Author Contributions
Funding
Acknowledgments
Conflicts of Interest
References
- O’Dell, B.L.; De Boland, A.R.; Koirtyohann, S.R. Distribution of phytate and nutritionally important elements among the morphological components of cereal grains. J. Agric. Food Chem. 1972, 20, 718–723. [Google Scholar] [CrossRef]
- Raboy, V.; Dickinson, D.B.; Neuffer, M.G. A survey of maize kernel mutants for variation in phytic acid. Maydica 1990, 35, 383–390. [Google Scholar]
- Raboy, V. Accumulation and Storage of Phosphate and Minerals. In Cellular and Molecular Biology of Plant Seed Development; Larkins, B.A., Vasil, I.K., Eds.; Springer: Dordrecht, The Netherlands, 1997; Volume 4, pp. 441–477. [Google Scholar]
- Raboy, V. Progress in Breeding Low Phytate Crops. J. Nutr. 2002, 132, 503–505. [Google Scholar] [CrossRef] [PubMed]
- Laboure, A.M.; Gagnon, J.; Lescure, A.M. Purification and characterization of a phytase (myo-inositol-hexakisphosphate phosphohydrolase) accumulated in maize (Zea mays) seedlings during germination. Biochem. J. 1993, 295, 413–419. [Google Scholar] [CrossRef] [PubMed]
- Graf, E.; Eaton, J.W. Antioxidant functions of phytic acid. Free Radic. Biol. Med. 1990, 8, 61–69. [Google Scholar] [CrossRef]
- Doria, E.; Galleschi, L.; Calucci, L.; Pinzino, C.; Pilu, R.; Cassani, E.; Nielsen, E. Phytic acid prevents oxidative stress in seeds: Evidence from a maize (Zea mays L.) low phytic acid mutant. J. Exp. Bot. 2009, 60, 967–978. [Google Scholar] [CrossRef] [PubMed]
- Sharpley, A.; Meyer, M. Minimizing Agricultural Nonpoint-Source Impacts: A Symposium Overview. J. Environ. Qual. 1994, 23, 1–3. [Google Scholar] [CrossRef]
- Mendoza, C.; Viteri, F.E.; Lönnerdal, B.; Young, K.A.; Raboy, V.; Brown, K.H. Effect of genetically modified, low-phytic acid maize on absorption of iron from tortillas. Am. J. Clin. Nutr. 1998, 68, 1123–1127. [Google Scholar] [CrossRef]
- Hambidge, K.M.; Huffer, J.W.; Raboy, V.; Grunwald, G.K.; Westcott, J.L.; Sian, L.; Miller, L.V.; Dorsch, J.A.; Krebs, N.F. Zinc absorption from low-phytate hybrids of maize and their wild-type isohybrids. Am. J. Clin. Nutr. 2004, 79, 1053–1059. [Google Scholar] [CrossRef] [Green Version]
- Hambidge, K.M.; Krebs, N.F.; Westcott, J.L.; Sian, L.; Miller, L.V.; Peterson, K.L.; Raboy, V. Absorption of calcium from tortilla meals prepared from low-phytate maize. Am. J. Clin. Nutr. 2005, 82, 84–87. [Google Scholar] [CrossRef]
- Sparvoli, F.; Cominelli, E. Seed Biofortification and phytic acid reduction: A conflict of interest for the plant? Plants 2015, 4, 728–755. [Google Scholar] [CrossRef] [PubMed]
- Panzeri, D.; Cassani, E.; Doria, E.; Tagliabue, G.; Forti, L.; Campion, B.; Bollini, R.; Brearley, C.A.; Pilu, R.; Nielsen, E.; et al. A defective ABC transporter of the MRP family, responsible for the bean lpa1 mutation, affects the regulation of the phytic acid pathway, reduces seed myo-inositol and alters ABA sensitivity. New Phytol. 2011, 191, 70–83. [Google Scholar] [CrossRef] [PubMed]
- Naidoo, R.; Watson, G.M.F.; Derera, J.; Tongoona, P.; Laing, M.D. Marker-assisted selection for low phytic acid (lpa1-1) with single nucleotide polymorphism marker and amplified fragment length polymorphisms for background selection in a maize backcross breeding programme. Mol. Breed. 2012, 30, 1207–1217. [Google Scholar] [CrossRef]
- Sureshkumar, S.; Tamilkumar, P.; Senthil, N.; Nagarajan, P.; Thangavelu, A.U.; Raveendran, M.; Vellaikumar, S.; Ganesan, K.N.; Balagopal, R.; Vijayalakshmi, G.; et al. Marker assisted selection of low phytic acid trait in maize (Zea mays L.). Hereditas 2014, 151, 20–27. [Google Scholar] [CrossRef] [PubMed]
- Raboy, V.; Gerbasi, P.F.; Young, K.A.; Stoneberg, S.D.; Pickett, S.G.; Bauman, A.T.; Murthy, P.P.N.; Sheridan, W.F.; Ertl, D.S. Origin and seed phenotype of maize low phytic acid 1-1 and low phytic acid 2-1. Plant Physiol. 2000, 124, 355–368. [Google Scholar] [CrossRef] [PubMed]
- Pilu, R.; Panzeri, D.; Gavazzi, G.; Rasmussen, S.K.; Consonni, G.; Nielsen, E. Phenotypic, genetic and molecular characterization of a maize low phytic acid mutant (lpa241). Theor. Appl. Genet. 2003, 107, 980–987. [Google Scholar] [CrossRef] [PubMed]
- Shi, J.; Wang, H.; Hazebroek, K.; Ertl, D.S.; Harp, T. The maize low-phytic acid 3 encodes a myo-inositol kinase that plays a role in phytic acid biosynthesis in developing seeds. Plant J. 2005, 42, 708–719. [Google Scholar] [CrossRef]
- Larson, S.R.; Young, K.A.; Cook, A.; Blake, T.K.; Raboy, V. Linkage mapping of two mutations that reduce phytic acid content of barley grain. Theor. Appl. Genet. 1998, 97, 141–146. [Google Scholar] [CrossRef]
- Rasmussen, S.K.; Hatzack, F. Identification of two low-phytate barley (Hordeum vulgare L.) grain mutants by TLC and genetic analysis. Hereditas 1998, 129, 107–112. [Google Scholar] [CrossRef]
- Bregitzer, P.; Raboy, V. Effects of four independent low-phytate mutations in barley (Hordeum vulgare L.) on seed phosphorus characteristics and malting quality. Cereal Chem. 2006, 83, 460–464. [Google Scholar] [CrossRef]
- Guttieri, M.; Bowen, D.; Dorsch, J.A.; Raboy, V.; Souza, E. Identification and characterization of a low phytic acid wheat. Crop Sci. 2004, 44, 418–424. [Google Scholar] [CrossRef]
- Liu, Q.L.; Xu, X.H.; Ren, X.L.; Fu, H.W.; Wu, D.X.; Shu, Q.Y. Generation and characterization of low phytic acid germplasm in rice (Oryza sativa L.). Theor. Appl. Genet. 2007, 114, 803–814. [Google Scholar] [CrossRef] [PubMed]
- Wilcox, J.R.; Premachandra, G.S.; Young, K.A.; Raboy, V. Isolation of high seed inorganic P, low-phytate soybean mutants. Crop Sci. 2000, 40, 1601–1605. [Google Scholar] [CrossRef]
- Hitz, W.D.; Carlson, T.J.; Kerr, P.S.; Sebastian, S.A. Biochemical and molecular characterization of a mutation that confers a decreased raffinosaccharide and phytic acid phenotype on soybean seeds. Plant Physiol. 2002, 128, 650–660. [Google Scholar] [CrossRef] [PubMed]
- Yuan, F.J.; Zhao, H.J.; Ren, X.L.; Zhu, S.L.; Fu, X.J.; Shu, Q.Y. Generation and characterization of two novel low phytate mutations in soybean (Glycine max L. Merr.). Theor. Appl. Genet. 2007, 115, 945–957. [Google Scholar] [CrossRef] [PubMed]
- Campion, B.; Sparvoli, F.; Doria, E.; Tagliabue, G.; Galasso, I.; Fileppi, M.; Bollini, R.; Nielsen, E. Isolation and characterisation of an lpa (low phytic acid) mutant in common bean (Phaseolus vulgaris L.). Theor. Appl. Genet. 2009, 118, 1211–1221. [Google Scholar] [CrossRef] [PubMed]
- Chen, P.S.; Toribara, T.Y.; Warner, H. Microdetermination of phosphorus. Anal. Chem. 1956, 28, 1756–1758. [Google Scholar] [CrossRef]
- Shi, J.; Wang, H.; Schellin, K.; Li, B.; Faller, M.; Stoop, J.M.; Meeley, R.B.; Ertl, D.E.; Ranch, J.P.; Glassman, K. Embryo-specific silencing of a transporter reduces phytic acid content of maize and soybean seeds. Nat. Biotechnol. 2007, 25, 930–937. [Google Scholar] [CrossRef]
- Swarbreck, D.; Ripoll, P.J.; Brown, D.A.; Edwards, K.J.; Theodoulou, F. Isolation and characterisation of two multidrug resistance associated protein genes from maize. Gene 2003, 315, 153–164. [Google Scholar] [CrossRef]
- Klein, M.; Burla, B.; Martinoia, E. The multidrug resistance-associated protein (MRP/ABCC) subfamily of ATP-binding cassette transporters in plants. FEBS Lett. 2006, 580, 1112–1122. [Google Scholar] [CrossRef]
- Maes, T.; De Keukeleire, P.; Gerats, T. Plant tagnology. Trends Plant Sci. 1999, 4, 90–96. [Google Scholar] [CrossRef]
- Vollbrecht, E.; Duvick, J.; Schares, J.P.; Ahern, K.R.; Deewatthanawong, P.; Xu, L.; Conrad, L.J.; Kikuchi, K.; Kubinec, T.A.; Hall, B.D.; et al. Genome-wide distribution of transposed Dissociation elements in maize. Plant Cell 2010, 22, 1667–1685. [Google Scholar] [CrossRef] [PubMed]
- Dooner, H.K.; Belachew, A. Transposition pattern of the maize element Ac from the Bz-M2(ac) allele. Genetics 1989, 122, 447–457. [Google Scholar] [PubMed]
- Landoni, M.; Badone, F.C.; Haman, N.; Schiraldi, A.; Fessas, F.; Cesari, V.; Toschi, I.; Cremona, R.; Delogu, C.; Villa, D.; et al. Low Phytic Acid 1 mutation in maize modifies density, starch properties, cations, and fiber contents in the seed. J. Agric. Food Chem. 2013, 61, 4622–4630. [Google Scholar] [CrossRef] [PubMed]
- Cerino Badone, F.; Amelotti, M.; Cassani, E.; Pilu, R. Study of low phytic acid1-7 (lpa1-7), a new ZmMRP4 mutation in maize. J. Hered. 2012, 103, 598–605. [Google Scholar] [CrossRef] [PubMed]
- Honke, J.; Kozłowska, H.; Vidal-Valverde, C.; Frias, J.; Górecki, R. Changes in quantities of inositol phosphates during maturation and germination of legume seeds. Eur. Food Res. Technol. 1998, 206, 279–283. [Google Scholar] [CrossRef]
- Loewus, F.A.; Murthy, P.P.N. Myo-Inositol metabolism in plants. Plant Sci. 2000, 150, 1–19. [Google Scholar] [CrossRef]
- Davidsson, L.; Almgren, A.; Juillerat, M.A.; Hurrell, R.F. Manganese absorption in humans: The effect of phytic acid and ascorbic acid in soy formula. Am. J. Clin. Nutr. 1995, 62, 984–987. [Google Scholar] [CrossRef]
- Spencer, J.D.; Allee, G.L.; Sauber, T.E. Phosphorus bioavailability and digestibility of normal and genetically modified low-phytate corn for pigs. J. Anim. Sci. 2000, 78, 675–681. [Google Scholar] [CrossRef]
- Veum, T.L.; Ledoux, D.R.; Raboy, V.; Ertl, D.S. Low-phytic acid corn improves nutrient utilization for growing pigs. J. Anim. Sci. 2001, 79, 2873–2880. [Google Scholar] [CrossRef]
- Bohlke, R.A.; Thaler, R.C.; Stein, H.H. Calcium, phosphorus, and amino acid digestibility in low-phytate corn, normal corn, and soybean meal by growing pigs. J. Anim. Sci. 2005, 83, 2396–2403. [Google Scholar] [CrossRef] [PubMed]
- Chen, R.; Xue, G.; Chen, P.; Yao, B.; Yang, W.; Ma, Q.; Fan, Y.; Zhao, Z.; Tarczynski, M.C.; Shi, J. Transgenic maize plants expressing a fungal phytase gene. Transg. Res. 2008, 17, 633–643. [Google Scholar] [CrossRef] [PubMed]
- Golovan, S.P.; Meidinger, R.G.; Ajakaiye, A.; Cottrill, M.; Wiederkehr, M.Z.; Barney, D.J.; Plante, C.; Pollard, J.W.; Fan, M.Z.; Hayes, M.A.; et al. Pigs expressing salivary phytase produce low-phosphorus manure. Nat. Biotechnol. 2001, 19, 741–745. [Google Scholar] [CrossRef] [PubMed]
- Edwards, J.D.; Jackson, A.K.; McClung, A.M. Genetic architecture of grain chalk in rice and interactions with a low phytic acid locus. Field Crops Res. 2017, 205, 116–123. [Google Scholar] [CrossRef] [Green Version]
- Moreno, M.A.; Chen, J.; Greenblatt, I.; Dellaporta, S.L. Reconstitutional mutagenesis of the maize P gene by short-range Ac transpositions. Genetics 1992, 131, 939–956. [Google Scholar] [PubMed]
- Pilu, R.; Panzeri, D.; Cassani, E.; Cerino Badone, F.; Landoni, M.; Nielsen, E. A paramutation phenomenon is involved in the genetics of maize low phytic acid1-241 (lpa1-241) trait. Heredity 2009, 102, 236–245. [Google Scholar] [CrossRef] [PubMed]
Code | Total Seeds Tested | Putative Mutant Seeds Isolated | Sucrose Solution Density (23 °C) |
---|---|---|---|
R3962/R3967 (1) | 138 | 0 | 1.218 |
R3962/R3967 (2) | 271 | 2 | 1.218 |
R3962/R3967 (3) | 108 | 0 | 1.218 |
R3962/R3967 (4) | 232 | 4 | 1.218 |
R3962/R3967 (5) | 246 | 12 | 1.218 |
R3962/R3967 (6) | 175 | 5 | 1.218 |
R3962/R3967 (7) | 132 | 3 | 1.218 |
R3963/R3967 (1) | 159 | 25 | 1.218 |
R3963/R3967 (2) | 196 | 1 | 1.218 |
R3963/R3967 (3) | 170 | 1 | 1.218 |
R3963/R3967 (4) | 101 | 14 | 1.218 |
R3963/R3970 (1) | 194 | 3 | 1.218 |
R3963/R3970 (2) | 143 | 7 | 1.218 |
R3964/R3967 (1) | 232 | 4 | 1.218 |
R3964/R3967 (2) | 258 | 4 | 1.218 |
R3964/R3967 (4) | 219 | 27 | 1.218 |
R3964/R3967 (5) | 232 | 20 | 1.218 |
R3964/R3967 (6) | 217 | 29 | 1.218 |
R3965/R3970 (2) | 160 | 23 | 1.218 |
R3966/R3970 (8) | 114 | 2 | 1.218 |
R3893/R916-300 | 166 | 8 | 1.218 |
R864/R916/(3) | 335 | 20 | 1.222 |
R864/R916 (5) | 285 | 14 | 1.222 |
R864/R916 (8) | 159 | 27 | 1.222 |
R864/R916 (9) | 145 | 16 | 1.222 |
Total | 4787 | 271 |
Code | N° of Seeds Tested | Phenotype | S/I % | |
---|---|---|---|---|
S/I | W/WT | |||
R4889 ⊗ | 24 | 13 | 11 | 54.17% |
R4890 ⊗ | 36 | 15 | 21 | 41.67% |
R4891 ⊗ | 36 | 8 | 28 | 22.22% |
R4892 ⊗ | 42 | 41 | 1 | 97.62% |
R4893 ⊗ | 18 | 17 | 1 | 94.44% |
R4894 ⊗ | 36 | 29 | 7 | 80.56% |
Total | 192 | 123 | 69 | 64.06% |
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Borlini, G.; Rovera, C.; Landoni, M.; Cassani, E.; Pilu, R. lpa1-5525: A New lpa1 Mutant Isolated in a Mutagenized Population by a Novel Non-Disrupting Screening Method. Plants 2019, 8, 209. https://doi.org/10.3390/plants8070209
Borlini G, Rovera C, Landoni M, Cassani E, Pilu R. lpa1-5525: A New lpa1 Mutant Isolated in a Mutagenized Population by a Novel Non-Disrupting Screening Method. Plants. 2019; 8(7):209. https://doi.org/10.3390/plants8070209
Chicago/Turabian StyleBorlini, Giulia, Cesare Rovera, Michela Landoni, Elena Cassani, and Roberto Pilu. 2019. "lpa1-5525: A New lpa1 Mutant Isolated in a Mutagenized Population by a Novel Non-Disrupting Screening Method" Plants 8, no. 7: 209. https://doi.org/10.3390/plants8070209