“…On the other hand, phenotypes of both FC1 and FC2 were the results of synergetic interactions between indigenous rhizobia (most likely including parasitic rhizobia), inoculants, genotype and other environmental factors. However, the BNF capacity of small nodules (diameter <2 mm) was significantly lower than that of big nodules (diameter >2 mm) (Li et al, 2018). Contrastingly, FC1 seemed to have a higher compatibility with the inoculated rhizobial strains than indigenous soil rhizobia as indicated by great increases of NN and NW after rhizobium inoculation, especially at low N level (Figure 1).…”
Soybean is an important economic and green manure crop that is widely used in intercropping and rotation systems due to its high biological nitrogen fixation (BNF) capacity and the resulting reduction in N fertilization. However, the genetic mechanisms underlying soybean BNF are largely unknown. Here, two soybean parent genotypes contrasting in BNF traits and 168 F9:11 recombinant inbred lines (RILs) were evaluated under four conditions in the field. The parent FC1 always produced more big nodules, yet fewer nodules in total than the parent FC2 in the field. Furthermore, nodulation in FC1 was more responsive to environmental changes than that in FC2. Broad-sense heritability (h2b) for all BNF traits varied from 0.48 to 0.87, which suggests that variation in the observed BNF traits was primarily determined by genotype. Moreover, two new QTLs for BNF traits, qBNF-16 and qBNF-17, were identified in this study. The qBNF-16 locus was detected under all of the four tested conditions, where it explained 15.9–59.0% of phenotypic variation with LOD values of 6.31–32.5. Meanwhile qBNF-17 explained 12.6–18.6% of observed variation with LOD values of 4.93–7.51. Genotype group analysis indicated that the FC1 genotype of qBNF-16 primarily affected nodule size (NS), while the FC2 genotype of qBNF-16 promoted nodule number (NN). On the other hand, the FC1 genotype of qBNF-17 influenced NN and the FC2 genotype of qBNF-17 impacted NS. The results on the whole suggest that these two QTLs might be valuable markers for breeding elite soybean varieties with high BNF capacities.
“…On the other hand, phenotypes of both FC1 and FC2 were the results of synergetic interactions between indigenous rhizobia (most likely including parasitic rhizobia), inoculants, genotype and other environmental factors. However, the BNF capacity of small nodules (diameter <2 mm) was significantly lower than that of big nodules (diameter >2 mm) (Li et al, 2018). Contrastingly, FC1 seemed to have a higher compatibility with the inoculated rhizobial strains than indigenous soil rhizobia as indicated by great increases of NN and NW after rhizobium inoculation, especially at low N level (Figure 1).…”
Soybean is an important economic and green manure crop that is widely used in intercropping and rotation systems due to its high biological nitrogen fixation (BNF) capacity and the resulting reduction in N fertilization. However, the genetic mechanisms underlying soybean BNF are largely unknown. Here, two soybean parent genotypes contrasting in BNF traits and 168 F9:11 recombinant inbred lines (RILs) were evaluated under four conditions in the field. The parent FC1 always produced more big nodules, yet fewer nodules in total than the parent FC2 in the field. Furthermore, nodulation in FC1 was more responsive to environmental changes than that in FC2. Broad-sense heritability (h2b) for all BNF traits varied from 0.48 to 0.87, which suggests that variation in the observed BNF traits was primarily determined by genotype. Moreover, two new QTLs for BNF traits, qBNF-16 and qBNF-17, were identified in this study. The qBNF-16 locus was detected under all of the four tested conditions, where it explained 15.9–59.0% of phenotypic variation with LOD values of 6.31–32.5. Meanwhile qBNF-17 explained 12.6–18.6% of observed variation with LOD values of 4.93–7.51. Genotype group analysis indicated that the FC1 genotype of qBNF-16 primarily affected nodule size (NS), while the FC2 genotype of qBNF-16 promoted nodule number (NN). On the other hand, the FC1 genotype of qBNF-17 influenced NN and the FC2 genotype of qBNF-17 impacted NS. The results on the whole suggest that these two QTLs might be valuable markers for breeding elite soybean varieties with high BNF capacities.
“…For histochemical analysis of GUS expression, soybean transgenic composite plants harboring miR169cPro:GUS and hairy root composite plants harboring GmNFYA‐CPro:GUS were inoculated with USDA110 under low N (LN, 50 μM) or high N (HN, 5000 μM) conditions. The procedures for GUS staining were performed as described previously (Li et al ., 2018).…”
Section: Methodsmentioning
confidence: 99%
“…The rhizobium inoculation was performed as described previously with few modifications (Li et al ., 2018). Briefly, soybean seeds were surface‐sterilized with 3% hydrogen peroxide (H 2 O 2 ) for 1 min and rinsed with distilled water, and then sowed in the sterilized vermiculite for seven days.…”
Summary
Legume crops contribute a great portion of clean nitrogen (N) to agro‐ecosystems through symbiotic N2 fixation in the nodule; however, the nodulation is always inhibited by high N availability which is known as the N inhibitory effect through largely unknown mechanisms.
We functionally investigated miR169c‐GmNFYA‐C‐GmENOD40 under multiple N conditions in soybean (Glycine max) (ENOD, Early Nodulin; NFYA, Nuclear Factor‐Y Subunit A). We elucidated their regulatory roles in soybean nodulation through analyzing expression patterns, micro–messenger RNA (miRNA–mRNA) interactions, phenotypes of transgenic soybean plants and genetic interactions.
We found that miR169c expression was induced by high N, whereas its target GmNFYA‐C was preferentially expressed in nodules and induced by rhizobium inoculation. Overexpression of miR169c inhibited nodulation through targeting 3′‐UTR of GmNFYA‐C, whereas knockout miR169c through CRISPR‐cas9 promoted nodulation. However, overexpression of GmNFYA‐C promoted soybean nodulation through facilitating rhizobium infection and increasing the expression of symbiotic signaling gene GmENOD40. Besides, GmNFYA‐C directly induced the expression of GmENOD40. In addition, overexpression of GmNFYA‐C without the target site of miR169c partially attenuated the inhibitory effect of high N on soybean nodulation.
We discovered a new regulatory pathway involving the miR169c‐NFYA‐C‐ENOD40 module that regulates soybean nodulation in response to N availability. This pathway provides substantial new insights into the mechanisms underlying the N inhibitory effect on nodulation.
“…In contrast, the excessive growth of nodules would lead to loss of yield because the balance would be disturbed [42]. In this context, Li et al [43] showed that an overexpression of the gene GmINS1 leads to an increase in the nodule number, biomass, and nitrogenase activity of large nodules. Peng et al [44] observed an increase in nodular mass when magnesium was added under nitrogen-limited conditions, as well as an improved N 2 fixation performance of the nodules, which can be attributed to a changed carbohydrate distribution.…”
Section: Plant Growth and Nutrient Concentrationmentioning
Soil samples from different locations with varied soybean cultivation histories were taken from arable fields in 2018 in East Germany and Poland (Lower Silesia) to evaluate the specific microsymbionts of the soybean, Bradyrhizobium japonicum, one to seven years after inoculation. Soybeans were grown in the selected farms between 2011 and 2017. The aim of the experiment was to investigate whether there is a difference in rhizobia contents in soils in which soybeans have been recultivated after one to seven years break, and whether this could lead to differences in soybean plant growth. The obtained soil samples were directly transferred into containers, then sterilized soybean seeds were sown into pots in the greenhouse. After 94 days of growth, the plants were harvested and various parameters such as the nodular mass, number of nodules, and dry matter in the individual plant parts were determined. In addition, the relative abundances of Bradyrhizobium sp. in soil samples were identified by sequencing. No major decline in Bradyrhizobium sp. concentration could be observed due to a longer interruption of soybean cultivation. Soil properties such as pH, P, and Mg contents did not show a significant influence on the nodule mass or number, but seem to have an influence on the relative abundance of Bradyrhizobium sp. The investigations have shown that Bradyrhizobium japonicum persists in arable soils even under Central European site conditions and enters into an effective symbiosis with soybeans for up to seven years.
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