Overexpression of the StP5CS gene promotes nodulation and nitrogen fixation in vegetable soybean under drought stress

2019 ◽  
Author(s):  
X. X. Wang ◽  
F. . Gao ◽  
S. P. Yang ◽  
J. Y. Gai ◽  
Y. L. Zhu
Keyword(s):  
Nitrogen Fixation ◽  
Drought Stress ◽  
Symbiotic Nitrogen Fixation ◽  
Nodule Development ◽  
Vegetable Soybean ◽  
Wild Type ◽  
Total N ◽  
Transgenic Lines ◽  
Glycine Max L ◽  
Better Than

The StP5CS (GenBank accession number: JN606861) T6 homozygous transgenic lines (HTLs) of vegetable soybean [Glycine max (L.) Merrill] were grown using vermiculite pot culture to determine whether StP5CS overexpression would enhance nodulation and symbiotic nitrogen fixation (SNF) in two T6 HTLs (17W-1, 17W-2) under drought conditions. The growth performance, nodule development and seed weight of T6 HTLs were significantly better than those of wild type (WT) plants. The proline levels in various tissues of T6 HTLs were higher than WT plants. The concentrations of total ureide, total N, leghemoglobin (Lb) and the activity of glutamine synthetase (GS, EC 6.3.1.2) in the T6 HTLs were significantly increased. Moreover, the relative expression levels of five key nodulation- and SNF-associated genes (i.e., GmENOD40-1, GmENOD40-2, GmLba, GmGS1â1 and GmGS1â2) were significantly higher in T6 HTLs. In conclusion, overexpression of StP5CS enhances nodulation and SNF in transgenic vegetable soybean under drought stress conditions

Starch content and activities of starch-metabolizing enzymes in effective and ineffective root nodules of soybean

1991 ◽  
Vol 69 (4) ◽  
pp. 697-701 ◽  
Author(s):  
Sharon I. Forrest ◽  
Desh Pal S. Verma ◽  
Rajinder S. Dhindsa
Keyword(s):  
Nitrogen Fixation ◽  
Starch Content ◽  
Root Nodules ◽  
Starch Synthase ◽  
Nodule Development ◽  
Wild Type ◽  
Starch Phosphorylase ◽  
High Concentration ◽  
Key Words Nitrogen Fixation ◽  
Glycine Max L

Starch content and activities of some enzymes of starch metabolism were determined in wild-type, N2-fixing (fix+) nodules and in two non-N2-fixing (fix−) nodules induced by Bradyrhizobium japonicum mutant strains, T5-95 and T8-1, on soybean (Glycine max L.) roots. The T5-95 nodules are similar to wild type in ultrastructure, but the T8-1 nodules are different in that the bacteroids are not released from the infection thread. After initial accumulation to relatively high concentration, starch was depleted during nitrogen fixation in fix+ nodules. However, in fix− nodules, the accumulated starch was not metabolized. The activity of starch-bound starch synthase (EC 2.4.1.21) declined in fix+ nodules but remained high in fix− nodules. The activity of α-amylase (EC 3.2.1.1) was only slightly higher than wild type in T5-95 but was four times higher than wild type in T8-1 nodules. The activity of starch phosphorylase (EC 2.4.1.1) increased in all nodule types from 14 to 21 days postinfection. A positive correlation was observed between the capacity of nodules to fix N2 and their capacity to degrade starch. Collectively, these results support the concept that starch accumulated during early stages of nodule development is metabolized to supply energy for nitrogen fixation and to meet the metabolic demands of bacteroids. Key words: nitrogen fixation, starch content, effective and ineffective nodules, starch synthase, starch phosphorylase, α-amylase.

Differential responses of the sunn4 and rdn1-1 super-nodulation mutants of Medicago truncatula to elevated atmospheric CO2

2021 ◽  
Author(s):  
Yunfa Qiao ◽  
Shujie Miao ◽  
Jian Jin ◽  
Ulrike Mathesius ◽  
Caixian Tang
Keyword(s):  
Nitrogen Fixation ◽  
Elevated Co2 ◽  
Medicago Truncatula ◽  
N2 Fixation ◽  
Sink Strength ◽  
Wild Type ◽  
Total N ◽  
Autoregulation Of Nodulation ◽  
Nodulation Mutants ◽  
Fixation Capacity

Abstract Background and Aims Nitrogen fixation in legumes requires tight control of carbon and nitrogen balance. Thus, legumes control nodule numbers via an autoregulation mechanism. ‘Autoregulation of nodulation’ mutants super-nodulate and are thought to be carbon-limited due to the high carbon-sink strength of excessive nodules. This study aimed to examine the effect of increasing carbon supply on the performance of super-nodulation mutants. Methods We compared the responses of Medicago truncatula super-nodulation mutants (sunn-4 and rdn1-1) and wild type to five CO2 levels (300-850 μmol mol -1). Nodule formation and N2 fixation were assessed in soil-grown plants at 18 and 42 days after sowing. Key results Shoot and root biomass, nodule number and biomass, nitrogenase activity and fixed-N per plant of all genotypes increased with increasing CO2 concentration and reached the maximum around 700 μmol mol -1. While the sunn-4 mutant showed strong growth-retardation compared to wild-type plants, elevated CO2 increased shoot biomass and total N content of rdn1-1 mutant up to two-fold. This was accompanied by a four-fold increase in nitrogen fixation capacity in the rdn1-1 mutant. Conclusions These results suggest that the super-nodulation phenotype per se did not limit growth. The additional nitrogen fixation capacity of the rdn1-1 mutant may enhance the benefit of elevated CO2 on plant growth and N2 fixation.

scholarly journals Antioxidant ability of glutaredoxins and their role in symbiotic nitrogen fixation in Rhizobium leguminosarum bv. viciae 3841

2020 ◽  
Author(s):  
Qian Zou ◽  
Yanlin Zhou ◽  
Guojun Cheng ◽  
Yang Peng ◽  
Sha Luo ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Quantitative Proteomics ◽  
Rhizobium Leguminosarum ◽  
Symbiotic Nitrogen Fixation ◽  
Proteome Analysis ◽  
Wild Type ◽  
Triple Mutant ◽  
Antioxidant Proteins ◽  
Mixed Disulfides ◽  
Nodule Symbiosis

Glutaredoxins (Grx) are redoxin family proteins that reduce disulfides and mixed disulfides between glutathione and proteins. Rhizobium leguminosarum bv. Viciae 3841 contains three genes coding for glutaredoxins: RL4289 (grxA) codes for a dithiolic glutaredoxin, RL2615 (grxB) codes for a monothiol glutaredoxin, while RL4261 (grxC) codes for a glutaredoxin-like NrdH protein. We generated mutants interrupted in one, two, or three glutaredoxin genes. These mutants had no obvious differences in growth phenotypes from the wild type RL3841. However, while a mutant of grxC did not affect the antioxidant or symbiotic capacities of R. leguminosarum, grxA-derived or grxB mutants decreased antioxidant and nitrogen fixation capacities. Furthermore, grxA mutants were severely impaired in rhizosphere colonization, and formed smaller nodules with defects of bacteroid differentiation, whereas nodules induced by grxB mutants contained abnormally thick cortices and prematurely senescent bacteroids. The grx triple mutant had the greatest defect in antioxidant and symbiotic capacities of R. leguminosarum and quantitative proteomics revealed it had 56 up-regulated and 81 down-regulated proteins relative to wildtype. Of these proteins, twenty-eight are involved in transporter activity, twenty are related to stress response and virulence, and sixteen are involved in amino acid metabolism. Overall, R. leguminosarum glutaredoxins behave as antioxidant proteins mediating root nodule symbiosis. IMPORTANCE Glutaredoxin catalyzes glutathionylation/deglutathionylation reactions, protects SH-groups from oxidation and restores functionally active thiols. Three glutaredoxins exist in R. leguminosarum and their properties were investigated in free-living bacteria and during nitrogen-fixing symbiosis. All the glutaredoxins were necessary for oxidative stress defense. Dithiol GrxA affects nodulation and nitrogen fixation of bacteroids by altering deglutathionylation reactions, monothiol GrxB is involved in symbiotic nitrogen fixation by regulating Fe-S cluster biogenesis, and GrxC may participate in symbiosis by an unknown mechanism. Proteome analysis provides clues to explain the differences between the grx triple mutant and wild-type nodules.

scholarly journals CsCYT75B1, a Citrus CYTOCHROME P450 Gene, Is Involved in Accumulation of Antioxidant Flavonoids and Induces Drought Tolerance in Transgenic Arabidopsis

Antioxidants ◽  
2020 ◽  
Vol 9 (2) ◽  
pp. 161 ◽  
Author(s):  
Muhammad Junaid Rao ◽  
Yuantao Xu ◽  
Xiaomei Tang ◽  
Yue Huang ◽  
Jihong Liu ◽  
...  
Keyword(s):  
Cytochrome P450 ◽  
Drought Stress ◽  
Drought Tolerance ◽  
Transgenic Arabidopsis ◽  
Wild Type ◽  
Ros Scavenging ◽  
Cytochrome P450 Gene ◽  
P450 Gene ◽  
Large Gene ◽  
Transgenic Lines

CYTOCHROME P450s genes are a large gene family in the plant kingdom. Our earlier transcriptome data revealed that a CYTOCHROME P450 gene of Citrus sinensis (CsCYT75B1) was associated with flavonoid metabolism and was highly induced after drought stress. Here, we characterized the function of CsCYT75B1 in drought tolerance by overexpressing it in Arabidopsis thaliana. Our results demonstrated that the overexpression of the CsCYT75B1 gene significantly enhanced the total flavonoid contents with increased antioxidant activity in transgenic Arabidopsis. The gene expression results showed that several genes that are responsible for the biosynthesis of antioxidant flavonoids were induced by 2–12 fold in transgenic Arabidopsis lines. After 14 days of drought stress, all transgenic lines displayed an enhanced tolerance to drought stress along with accumulating antioxidant flavonoids with lower superoxide radicals and reactive oxygen species (ROS) than wild type plants. In addition, drought-stressed transgenic lines possessed higher antioxidant enzymatic activities than wild type transgenic lines. Moreover, the stressed transgenic lines had significantly lower levels of electrolytic leakage than wild type transgenic lines. These results demonstrate that the CsCYT75B1 gene of sweet orange functions in the metabolism of antioxidant flavonoid and contributes to drought tolerance by elevating ROS scavenging activities.

scholarly journals Digalactosyldiacylglycerol Synthase Gene MtDGD1 Plays an Essential Role in Nodule Development and Nitrogen Fixation

2019 ◽  
Vol 32 (9) ◽  
pp. 1196-1209
Author(s):  
Zaiyong Si ◽  
Qianqian Yang ◽  
Rongrong Liang ◽  
Ling Chen ◽  
Dasong Chen ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Essential Role ◽  
Symbiotic Nitrogen Fixation ◽  
Histochemical Staining ◽  
Nodule Development ◽  
Nodule Number ◽  
Nitrogen Fixation Activity ◽  
Fixation Activity ◽  
Infected Cells ◽  
Nodule Symbiosis

Little is known about the genes participating in digalactosyldiacylglycerol (DGDG) synthesis during nodule symbiosis. Here, we identified full-length MtDGD1, a synthase of DGDG, and characterized its effect on symbiotic nitrogen fixation in Medicago truncatula. Immunofluorescence and immunoelectron microscopy showed that MtDGD1 was located on the symbiosome membranes in the infected cells. β-Glucuronidase histochemical staining revealed that MtDGD1 was highly expressed in the infection zone of young nodules as well as in the whole mature nodules. Compared with the control, MtDGD1-RNA interference transgenic plants exhibited significant decreases in nodule number, symbiotic nitrogen fixation activity, and DGDG abundance in the nodules, as well as abnormal nodule and symbiosome development. Overexpression of MtDGD1 resulted in enhancement of nodule number and nitrogen fixation activity. In response to phosphorus starvation, the MtDGD1 expression level was substantially upregulated and the abundance of nonphospholipid DGDG was significantly increased in the roots and nodules, accompanied by corresponding decreases in the abundance of phospholipids such as phosphatidylcholine, phosphatidylethanolamine, and phosphatidylinositol. Overall, our results indicate that DGD1 contributes to effective nodule organogenesis and nitrogen fixation by affecting the synthesis and content of DGDG during symbiosis.

scholarly journals Succinate Transport Is Not Essential for Symbiotic Nitrogen Fixation by Sinorhizobium meliloti or Rhizobium leguminosarum

2017 ◽  
Vol 84 (1) ◽  
Author(s):  
Michael J. Mitsch ◽  
George C. diCenzo ◽  
Alison Cowie ◽  
Turlough M. Finan
Keyword(s):  
Nitrogen Fixation ◽  
Carbon Source ◽  
Sinorhizobium Meliloti ◽  
Rhizobium Leguminosarum ◽  
Symbiotic Nitrogen Fixation ◽  
Sequencing Analysis ◽  
Wild Type ◽  
Content Type ◽  
Transcriptional Changes ◽  
Symbiotic Properties

ABSTRACTSymbiotic nitrogen fixation (SNF) is an energetically expensive process performed by bacteria during endosymbiotic relationships with plants. The bacteria require the plant to provide a carbon source for the generation of reductant to power SNF. While C4-dicarboxylates (succinate, fumarate, and malate) appear to be the primary, if not sole, carbon source provided to the bacteria, the contribution of each C4-dicarboxylate is not known. We address this issue using genetic and systems-level analyses. Expression of a malate-specific transporter (MaeP) inSinorhizobium melilotiRm1021dctmutants unable to transport C4-dicarboxylates resulted in malate import rates of up to 30% that of the wild type. This was sufficient to support SNF withMedicago sativa, with acetylene reduction rates of up to 50% those of plants inoculated with wild-typeS. meliloti.Rhizobium leguminosarumbv. viciae 3841dctmutants unable to transport C4-dicarboxylates but expressing themaePtransporter had strong symbiotic properties, withPisum sativumplants inoculated with these strains appearing similar to plants inoculated with wild-typeR. leguminosarum. This was despite malate transport rates by the mutant bacteroids being 10% those of the wild type. An RNA-sequencing analysis of the combinedP. sativum-R. leguminosarumnodule transcriptome was performed to identify systems-level adaptations in response to the inability of the bacteria to import succinate or fumarate. Few transcriptional changes, with no obvious pattern, were detected. Overall, these data illustrated that succinate and fumarate are not essential for SNF and that, at least in specific symbioses,l-malate is likely the primary C4-dicarboxylate provided to the bacterium.IMPORTANCESymbiotic nitrogen fixation (SNF) is an economically and ecologically important biological process that allows plants to grow in nitrogen-poor soils without the need to apply nitrogen-based fertilizers. Much research has been dedicated to this topic to understand this process and to eventually manipulate it for agricultural gains. The work presented in this article provides new insights into the metabolic integration of the plant and bacterial partners. It is shown that malate is the only carbon source that needs to be available to the bacterium to support SNF and that, at least in some symbioses, malate, and not other C4-dicarboxylates, is likely the primary carbon provided to the bacterium. This work extends our knowledge of the minimal metabolic capabilities the bacterium requires to successfully perform SNF and may be useful in further studies aiming to optimize this process through synthetic biology approaches. The work describes an engineering approach to investigate a metabolic process that occurs between a eukaryotic host and its prokaryotic endosymbiont.

scholarly journals Induction and Spatial Organization of Polyamine Biosynthesis During Nodule Development in Lotus japonicus

2004 ◽  
Vol 17 (12) ◽  
pp. 1283-1293 ◽  
Author(s):  
Emmanouil Flemetakis ◽  
Rodica C. Efrose ◽  
Guilhem Desbrosses ◽  
Maria Dimou ◽  
Costas Delis ◽  
...  
Keyword(s):  
Nitrogen Fixation ◽  
Symbiotic Nitrogen Fixation ◽  
Expression Patterns ◽  
Lotus Japonicus ◽  
Cyclin D3 ◽  
Nodule Development ◽  
Cdna Clones ◽  
Long Distance ◽  
Polyamine Biosynthesis ◽  
Transcript Levels

Putrescine and other polyamines are produced by two alternative pathways in plants. One pathway starts with the enzyme arginine decarboxylase (ADC; EC 4.1.1.19), the other with ornithine decarboxylase (ODC; EC 4.1.1.17). Metabolite profiling of nitrogen-fixing Lotus japonicus nodules, using gas chromatography coupled to mass spectrometry, revealed a two- to sixfold increase in putrescine levels in mature nodules compared with other organs. Genes involved in polyamine biosynthesis in L. japonicus nodules were identified by isolating cDNA clones encoding ADC (LjADC1) and ODC (LjODC) from a nodule library. Searches of the public expressed sequence tag databases revealed the presence of a second gene encoding ADC (LjADC2). Real-time reverse-transcription-polymerase chain reaction analysis showed that LjADC1 and LjADC2 were expressed throughout the plant, while LjODC transcripts were detected only in nodules and roots. Induction of LjODC and LjADC gene expression during nodule development preceded symbiotic nitrogen fixation. Transcripts accumulation was maximal at 10 days postinfection, when a 6.5-fold increase in the transcript levels of LjODC was observed in comparison with the uninfected roots, while a twofold increase in the transcript levels of LjADC1 and LjADC2 was detected. At later stages of nodule development, transcripts for ADC drastically declined, while in the case of ODC, transcript accumulation was higher than that in roots until after 21 days postinfection. The expression profile of genes involved in putrescine biosynthesis correlated well with the expression patterns of genes involved in cell division and expansion, including a L. japonicus Cyclin D3 and an α-expansin gene. Spatial localization of LjODC and LjADC1 gene transcripts in developing nodules revealed that both transcripts were expressed in nodule inner cortical cells and in the central tissue. High levels of LjADC1 transcripts were also observed in both nodule and connecting root vascular tissue, suggesting that putrescine and other polyamines may be subject to long-distance transport. Our results indicate that polyamines are primarily involved in physiological and cellular processes involved in nodule development, rather than in processes that support directly symbiotic nitrogen fixation and assimilation.

Metabolite Regulation of Phosphenolpyruvate Carboxylase in Legume Root Nodules

1996 ◽  
Vol 23 (4) ◽  
pp. 413 ◽  
Author(s):  
KC Woo ◽  
S Xu
Keyword(s):  
Nitrogen Fixation ◽  
Glycine Max ◽  
Vigna Unguiculata ◽  
Symbiotic Nitrogen Fixation ◽  
Feedback Inhibition ◽  
Root Nodules ◽  
Psophocarpus Tetragonolobus ◽  
Glycine Max L ◽  
Fructose 1,6 Bisphosphate ◽  
Metabolite Regulation

The effects of metabolic activators and inhibitors on phosphoenolpyruvate carboxylase (PEPC) activity were examined at pH 7 in partially purified enzyme from nodules of soybean (Glycine max (L.) Merr.), Psophocarpus tetragonolobus DC. and Vigna unguiculata ssp. sesquipedalis (L.) Verdc. Glucose 6-phosphate, fructose 6-phosphate, glucose 1-phosphate, fructose 1-phosphate, fructose 1,6- bisphosphate and phosphoglycerate stimulated the activity about 2-fold at low (0.5 mM) but not saturating (2.5 mM) PEP concentration. Glc 6-P and fru 6-P were the most effective activators and they increased the affinity of the enzyme for PEP by 2-4-fold. The dicarboxylates, malate, succinate, malonate, 2-oxoglutarate and aspartate inhibited PEPC activity. Malate was the most inhibitory, and strongly inhibited PEPC activity even at saturating PEP concentration. The Ki values for malate were 0.3-0.4 mM for soybean and P. tetragonolobus. However, glc 6-P and fru 6-P alleviated maiate inhibition and increased the Ki values by 11- to 28-fold in these two species. We propose that glc 6-P (fru 6-P) activates PEPC in a feedforward regulation and protects it against feedback inhibition by malate and thus coordinates the supply of photosynthate availability with malate synthesis required by the bacteroids to support symbiotic nitrogen fixation in nodules.

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