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Parra-Gonzalez L.B.,Agriaquaculture Nutritional Genomic Center | Aravena-Abarzua G.A.,Agriaquaculture Nutritional Genomic Center | Navarro-Navarro C.S.,Agriaquaculture Nutritional Genomic Center | Udall J.,Brigham Young University | And 5 more authors.
BMC Genomics | Year: 2012

Background: Yellow lupin (Lupinus luteus L.) is a minor legume crop characterized by its high seed protein content. Although grown in several temperate countries, its orphan condition has limited the generation of genomic tools to aid breeding efforts to improve yield and nutritional quality. In this study, we report the construction of 454-expresed sequence tag (EST) libraries, carried out comparative studies between L. luteus and model legume species, developed a comprehensive set of EST-simple sequence repeat (SSR) markers, and validated their utility on diversity studies and transferability to related species.Results: Two runs of 454 pyrosequencing yielded 205 Mb and 530 Mb of sequence data for L1 (young leaves, buds and flowers) and L2 (immature seeds) EST- libraries. A combined assembly (L1L2) yielded 71,655 contigs with an average contig length of 632 nucleotides. L1L2 contigs were clustered into 55,309 isotigs. 38,200 isotigs translated into proteins and 8,741 of them were full length. Around 57% of L. luteus sequences had significant similarity with at least one sequence of Medicago, Lotus, Arabidopsis, or Glycine, and 40.17% showed positive matches with all of these species. L. luteus isotigs were also screened for the presence of SSR sequences. A total of 2,572 isotigs contained at least one EST-SSR, with a frequency of one SSR per 17.75 kbp. Empirical evaluation of the EST-SSR candidate markers resulted in 222 polymorphic EST-SSRs. Two hundred and fifty four (65.7%) and 113 (30%) SSR primer pairs were able to amplify fragments from L. hispanicus and L. mutabilis DNA, respectively. Fifty polymorphic EST-SSRs were used to genotype a sample of 64 L. luteus accessions. Neighbor-joining distance analysis detected the existence of several clusters among L. luteus accessions, strongly suggesting the existence of population subdivisions. However, no clear clustering patterns followed the accession's origin.Conclusion: L. luteus deep transcriptome sequencing will facilitate the further development of genomic tools and lupin germplasm. Massive sequencing of cDNA libraries will continue to produce raw materials for gene discovery, identification of polymorphisms (SNPs, EST-SSRs, INDELs, etc.) for marker development, anchoring sequences for genome comparisons and putative gene candidates for QTL detection. © 2012 Parra-González et al.; licensee BioMed Central Ltd.


Ogura T.,Agriaquaculture Nutritional Genomic Center | Ogihara J.,Nihon University | Sunairi M.,Nihon University | Takeishi H.,Nihon University | And 5 more authors.
Proteomics | Year: 2014

Yellow lupin (Lupinus luteus L.) is a legume crop containing a large amount of protein in its seeds. In this study, we constructed a seed-protein catalog to provide a foundation for further study of the seeds. A total of 736 proteins were identified in 341 2DE spots by nano-LC-MS/MS. Eight storage proteins were found as multiple spots in the 2DE gels. The 736 proteins correspond to 152 unique proteins as shown by UniRef50 clustering. Sixty-seven of the 152 proteins were associated with KEGG-defined pathways. Of the remaining proteins, 57 were classified according to a GO term. The functions of the remaining 28 proteins have yet to be determined. This is the first yellow lupin seed-protein catalog, and it contains considerably more data than previously reported for white lupin (L. albus L.). © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.


Parra Gonzalez L.B.,Agro Aquaculture Nutritional Genomic Center | Straub S.C.K.,Cornell University | Doyle J.J.,Cornell University | Mora Ortega P.E.,Agro Aquaculture Nutritional Genomic Center | And 3 more authors.
American Journal of Botany | Year: 2010

Premise of the study: Microsatellite primers were developed in Lupinus luteus L., an emerging temperate protein crop, to investigate genetic diversity, population structure, and to facilitate the generation of better yellow lupine varieties. Methods and Results: Thirteen olymorphic primer sets were evaluated in a European and Eastern European accession collection of L. luteus. The primers amplified di-, tri-, and tetranucleotide repeats with 2-4 alleles per locus. These revealed a moderate to low level of genetic variation, as indicated by an average observed heterozygosity of 0.0126. Select loci also amplified successfully in the closely related species L. hispanicus Boiss. & Reut. and in the New World species L. mutabilis Sweet. Conclusions: These results indicate the utility of primers for the study of genetic diversity across L. luteus populations and related lupine species. The use of these microsatellite markers will facilitate the implementation of several molecular breeding strategies in yellow lupine. © 2010 Botanical Society of America.


Morales C.G.,Institute of Agricultural Research INIA | Pino M.T.,Institute of Agricultural Research INIA | del Pozo A.,University of Talca
Scientia Horticulturae | Year: 2013

Raspberry (Rubus idaeus L.) is a deciduous plant with perennial roots, 75% of which are concentrated in the upper level of the soil. Its shallow rooting system requires a regular water supply; a water deficit can affect fructification as well as cane growth and yield for the following season. Despite the demonstrated drought stress impact on the raspberry, there is little information about the phenological and physiological responses to drought stress. The main goal of this study was to evaluate the effects of drought stress on the phenological phases, physiological parameters and yield of two raspberry cultivars: Heritage (remontant type) and Meeker (non-remontant type). All plants were grown in pots under greenhouse conditions, and the following watering treatments were applied: (T1) well-watered, 100% irrigation and (T2) a controlled drought-stress cycle. The volumetric soil water content (θ), phenological phases, leaf net photosynthetic rate (A), transpiration rate (T), and stomatal conductance (gs) were registered periodically. The free proline and total soluble sugars were also determined. Based on the phenological study, Heritage under drought-stress (T2) showed earlier flowering and a shorter fruit production period in relation to well-watered plants (T1). In Meeker, T2 extended the cane and summer lateral elongation, showing earlier senescence. Leaf gas exchange decreased with drought stress, A declined after 28-day period under drought stress, from 9.2μmolCO2m-2s-1 to 3.0μmolCO2m-2s-1 in Heritage, and from 12.2μmolCO2m-2s-1 to 3.0μmolCO2m-2s-1 in Meeker. In both cultivars, the free proline and total soluble sugars increased with drought stress. The fruit production was also affected in the next season under T2 condition, decreasing in 34 and 38% in relation to well-watered plants. © 2013 The Authors.

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