RESUMO
The objective of this study was to use RNA interference (RNAi) to improve protein quality and decrease anti-nutritional effects in soybean. Agrobacterium tumefaciens-mediated transformation was conducted using RNAi and an expression vector containing the 7S globulin ß-subunit gene. The BAR gene was used as the selective marker and cotyledonary nodes of soybean genotype Jinong 27 were chosen as explant material. Regenerated plants were detected by molecular biology techniques. Transformation of the ß-subunit gene in the 7S protein was detected by PCR, Southern blot, and q-PCR. Positive plants (10 T0, and 6 T1, and 13 T2) were tested by PCR. Hybridization bands were detected by Southern blot analysis in two of the T1 transgenic plants. RNAi expression vectors containing the soybean 7S protein ß-subunit gene were successfully integrated into the genome of transgenic plants. qRT-PCR analysis in soybean seeds showed a clear decrease in expression of the soybean ß-subunit gene. The level of 7S protein ß-subunit expression in transgenic plants decreased by 77.5% as compared to that of the wild-type plants. This study has established a basis for the application of RNAi to improve the anti-nutritional effects of soybean.
Assuntos
Agrobacterium tumefaciens/genética , Antígenos de Plantas/genética , Globulinas/genética , Glycine max/genética , Interferência de RNA , Proteínas de Armazenamento de Sementes/genética , Proteínas de Soja/genética , Antígenos de Plantas/metabolismo , Cotilédone/citologia , Cotilédone/genética , Cotilédone/metabolismo , Técnicas de Transferência de Genes , Genoma de Planta , Globulinas/metabolismo , Recombinação Genética , Proteínas de Armazenamento de Sementes/metabolismo , Proteínas de Soja/metabolismo , TransgenesRESUMO
Plant traits are important indices for regulating and controlling yield ability in soybean varieties. It is important to comprehensively study the quantitative trait locus (QTL) mapping for soybean plant traits, cloning related genes, and marker assistant breeding. In this study, 236 F2 generation plants and a derivative group were constructed by using Jiyu50 and Jinong18, obtained from Jilin Province. A total of 102 simple sequence repeat markers were used to construct a genetic linkage map. With 2 years of molecular and phenotypic data, QTL analyses and mapping were conducted for soybean maturity, plant height, main stem node, main stem branch, seed weight per plant, and more. Five main plant traits were analyzed via inclusive composite interval mapping using QTL IciMapping v2.2. Using one-dimensional scanning, a total of 30 QTLs were detected and distributed across 1 (A1), 4 (C2), and 12 (G). There were 9 linkage groups, including 16 major QTLs. Using two-dimensional scanning, 7 pairs of epistatic QTL interactions for maturity and plant height were detected in the soybean.
Assuntos
Mapeamento Cromossômico/métodos , Glycine max/genética , Locos de Características Quantitativas , Cromossomos de Plantas/genética , DNA de Plantas/análise , Ligação Genética , Hibridização Genética , Repetições de MicrossatélitesRESUMO
Seed number per pod is an important component of yield traits in soybean (Glycine max L.). In 2010, we identified a natural mutant with an increased number of four-seed pods from a soybean variety named 'Jinong 18' (JN18). Subsequent observations indicated that the trait was stably inherited. To identify and understand the function of genes associated with this mutant trait, we analyzed the genetic differences between the mutant (JN18MT01) and source variety (JN18) by transcriptome sequencing. Three types of tissues, axillary buds, unfertilized ovaries, and young pods at three different growth stages, V6, R1, and R3, were analyzed, respectively. The sequencing results yielded 55,582 expressed genes and 4183 differentially expressed genes (DEGs). Among these, the log2 ratio value of 162 DEGs was >10, and 13 DEGs had overlapping expression at three different growth stages. Comparisons of DEGs among three different growth stages yielded similar results in terms of the percentage of genes classified into each gene ontology (GO) category. DEGs were classified into 25 different functional groups in clusters of orthologous groups analysis. Proportions of the main functional genes differed significantly over developmental stages. A comparison of enriched pathways among the three developmental stages revealed that 646 unigenes were involved in 103 metabolic pathways. These results show that the development of four-seed pods is associated with a complex network involving multiple physiological and metabolic pathways. This study lays the foundation for further research on cloning and on the molecular regulation of genes related to the four-seed pod mutation.
Assuntos
Frutas/genética , Regulação da Expressão Gênica de Plantas , Glycine max/genética , Proteínas de Plantas/genética , Característica Quantitativa Herdável , Sementes/genética , Transcriptoma , Frutas/anatomia & histologia , Frutas/crescimento & desenvolvimento , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Ontologia Genética , Redes Reguladoras de Genes , Sequenciamento de Nucleotídeos em Larga Escala , Redes e Vias Metabólicas/genética , Anotação de Sequência Molecular , Mutação , Fenótipo , Proteínas de Plantas/metabolismo , Sementes/crescimento & desenvolvimento , Glycine max/anatomia & histologia , Glycine max/crescimento & desenvolvimentoRESUMO
Here, we compare the molecular mechanism of soybean heterosis through the differential expression of basic cloning. Specifically, we cloned 22 differentially expressed cDNA fragments from hybrid combinations of Jilin 38 x EXP (which had obvious yield advantages) and their parents. In addition, we compared the homology of these fragments and predicted their functions. Cloning differentially expressed genes included the identification of the calmodulin binding protein, 18S ribosomal gene, 26S ribosomal gene, soybean satellite DNA, soybean acid phosphatase, soybean chlorophyll a/b-binding protein II (Cab-6) gene, soybean chloroplast PI 437654 gene, soybean PPR protein gene, and other fragments with unknown functions. In conclusion, the cloning and functional prediction of differentially expressed soybean genes in this study is anticipated to provide valuable information for studies on the molecular mechanism of heterosis.