Application of Synthetic Peptide CEP1 Increases Nutrient Uptake Rates Along Plant Roots

The root system of a plant provides vital functions including resource uptake, storage, and anchorage in soil. The uptake of macro-nutrients like nitrogen (N), phosphorus (P), potassium (K), and sulphur (S) from the soil is critical for plant growth and development. Small signaling peptide (SSP) hormones are best known as potent regulators of plant growth and development with a few also known to have specialized roles in macronutrient utilization. Here we describe a high throughput phenotyping platform for testing SSP effects on root uptake of multiple nutrients. The SSP, CEP1 (C-TERMINALLY ENCODED PEPTIDE) enhanced nitrate uptake rate per unit root length in Medicago truncatula plants deprived of N in the high-affinity transport range. Single structural variants of M. truncatula and Arabidopsis thaliana specific CEP1 peptides, MtCEP1D1:hyp4,11 and AtCEP1:hyp4,11, enhanced uptake not only of nitrate, but also phosphate and sulfate in both model plant species. Transcriptome analysis of Medicago roots treated with different MtCEP1 encoded peptide domains revealed that hundreds of genes respond to these peptides, including several nitrate transporters and a sulfate transporter that may mediate the uptake of these macronutrients downstream of CEP1 signaling. Likewise, several putative signaling pathway genes including LEUCINE-RICH REPEAT RECPTOR-LIKE KINASES and Myb domain containing transcription factors, were induced in roots by CEP1 treatment. Thus, a scalable method has been developed for screening synthetic peptides of potential use in agriculture, with CEP1 shown to be one such peptide.

Application of synthetic peptide CEP1 increases nutrient uptake rates along plant roots

The root system of a plant provides vital functions including resource uptake, storage, and anchorage in soil. Uptake from the soil of macro-nutrients like nitrogen (N), phosphorus (P), potassium (K), and sulphur (S) is critical for plant growth and development. Small signaling peptide (SSP) hormones are best known as potent regulators of plant growth and development with a few also known to have specialized roles in macronutrient utilization. Here we describe a high-throughput screen of SSP effects on root uptake of multiple nutrients. The SSP, MtCEP1 enhanced nitrate uptake rate per unit root length in Medicago truncatula plants deprived of N. MtCEP1 and AtCEP1 enhanced uptake not only of nitrate, but also phosphate and sulfate in both Medicago and Arabidopsis. Transcriptome analysis of Medicago roots treated with different MtCEP1 encoded peptide domains revealed that hundreds of genes respond to these peptides, including several nitrate transporters and a sulfate transporter that may mediate the uptake of these macronutrients downstream of CEP1 signaling. Likewise, several putative signaling pathway genes were induced in roots by CEP1 treatment. Thus, a scalable method has been developed for screening synthetic peptides of potential use in agriculture, with CEP1 shown to be one such peptide.

Initial evaluation of nitrogen uptake of Thalassia hemprichii in Nha Trang, Khanh Hoa

The uptake rate of nitrate by leaves and roots was simultaneously investigated in the seagrass T. hemprichii collected from Lo river, Nha Trang. The principle of nutrient uptake of seagrass was based on the approaches of the Michaelis-Menten model. The laboratory experiments of nitrate uptake were done the ranges of ambient nitrate concentrations in the separate water column of leaves and roots. The calculated results indicated that the nitrate uptake rate through the leaves was higher than that of the roots. The maximum uptake rate (Vmax) through the leaves and the roots was 1,777.0 mgN/g DW/h and 131.6 mgN/g DW/h, respectively. From these results, the nitrate uptake rate by the seagrass in Lo river was estimated at 52.2 mgNm-2h-1. The value plays an important role in developing and implementing plans for the protection and rehabilitation of seagrass beds in coastal areas.

Physiological response of a golden tide alga (<i>Sargassum muticum</i>) to the interaction of ocean acidification and phosphorus enrichment

The development of golden tides is potentially influenced by global change factors, such as ocean acidification and eutrophication, but related studies are very scarce. In this study, we cultured a golden tide alga, Sargasssum muticum, at two levels of pCO2 (400 and 1000 µatm) and phosphate (0.5 and 40 µM) to investigate the interactive effects of elevated pCO2 and phosphate on the physiological properties of the thalli. Higher pCO2 and phosphate (P) levels alone increased the relative growth rate by 41 and 48 %, the net photosynthetic rate by 46 and 55 %, and the soluble carbohydrates by 33 and 62 %, respectively, while the combination of these two levels did not promote growth or soluble carbohydrates further. The higher levels of pCO2 and P alone also enhanced the nitrate uptake rate by 68 and 36 %, the nitrate reductase activity (NRA) by 89 and 39 %, and the soluble protein by 19 and 15 %, respectively. The nitrate uptake rate and soluble protein was further enhanced, although the nitrate reductase activity was reduced when the higher levels of pCO2 and P worked together. The higher pCO2 and higher P levels alone did not affect the dark respiration rate of the thalli, but together they increased it by 32 % compared to the condition of lower pCO2 and lower P. The neutral effect of the higher levels of pCO2 and higher P on growth and soluble carbohydrates, combined with the promoting effect on soluble protein and dark respiration, suggests that more energy was drawn from carbon assimilation to nitrogen assimilation under conditions of higher pCO2 and higher P; this is most likely to act against the higher pCO2 that caused acid–base perturbation via synthesizing H+ transport-related protein. Our results indicate that ocean acidification and eutrophication may not boost golden tide events synergistically, although each one has a promoting effect.

Physiological response of a golden tide alga (<i>Sargassum muticum</i>) to the interaction of ocean acidification and phosphorus enrichment

The evolvement of golden tides would be influenced by global change factors, such as ocean acidification and eutrophication, but the related studies are very scarce. In this study, we cultured a golden tide alga, Sargasssum muticum, at two levels of pCO2 (400, 1000 µatm) and phosphate (0.5 µM, 40 µM) conditions to investigate the interactive effects of elevated pCO2 and phosphate on physiological properties of the thalli. The higher pCO2 level and phosphate (P) level alone increased the relative growth rate by 40.82 % and 47.78 %, net photosynthetic rate by 46.34 % and 55.16 %, soluble carbohydrates by 32.78 % and 61.83 % respectively whilst the combination of these two levels did not promote growth or soluble carbohydrates further. The higher levels of pCO2 and P alone also enhanced the nitrate uptake rate by 68.27 % and 35.89 %, nitrate reductase activity by 89.08 % and 39.31 %, and soluble protein by 19.05 % and 15.13 % respectively. The nitrate uptake rate and soluble protein was further enhanced although the nitrate reductase activity was reduced when the higher levels of pCO2 and P worked together. The higher pCO2 level and higher P level alone did not affect the dark respiration rate of thalli but they together increased it by 32.30 % compared to the condition of the lower pCO2 and lower P. The mute effect of the higher level of pCO2 and higher P on growth, soluble carbohydrates, combined with the promoting effect of it on soluble protein and dark respiration, suggests more energy was drawn from carbon assimilation to nitrogen assimilation at the condition of higher pCO2 and higher P, probably to act against the higher pCO2 caused acid-base perturbation via synthesizing H+ transport-related protein. Our results indicate ocean acidification and eutrophication may not boost the gold tides events synergistically although each of them alone has a promoting effect.

Uptake and Assimilation of Nitrate and Iron in Two Vaccinium Species as Affected by External Nitrate Concentration

Most Vaccinium species have narrow soil adaptation and are limited to soils that have low pH, high available iron (Fe), and nitrogen (N) primarily in the ammonium (NH4+) form. Vaccinium arboreum Marsh. is a wild species that can tolerate a wider range of soil conditions, including higher pH and nitrate (NO3-) as the predominant N form. This wider soil adaptation may be related to the ability of V. arboreum to acquire Fe and NO3- more efficiently than cultivated Vaccinium species, such as V. corymbosum L. interspecific hybrid (southern highbush). Nitrate and Fe uptake, and nitrate reductase (NR) and ferric chelate reductase (FCR) activities were compared in these two species grown hydroponically in either 1.0 or 5.0 mm NO3-. Nitrate uptake rate (on a whole-plant and FW basis) and root NR activity were significantly greater in V. arboreum compared with V. corymbosum. Iron uptake on a FW basis was also greater in V. arboreum, and was correlated with higher root FCR activity than was found in V. corymbosum. Increased Fe and NO3- uptake/assimilation in V. arboreum were reflected in increased organ and whole-plant dry weights compared with V. corymbosum. Vaccinium arboreum appears to be more efficient in acquiring and assimilating NO3- and Fe than is the cultivated species, V. corymbosum. This may partially explain the wider soil adaptation of V. arboreum.

A nitrate biosensor based methodology for monitoring anoxic activated sludge activity

An improved methodology based on a nitrate biosensor is developed and applied successfully for in-depth monitoring and study of anoxic activated sludge activities. The major advantages of the methodology are its simplicity, reliability and high data quality. The resulting data allowed for the first time to monitor anoxic respiration rate of activated sludge (nitrate uptake rate (NUR)) at a high time resolution making it clearly comparable with high frequency oxygen uptake rate (OUR) measurements obtained under aerobic conditions. Further, the anoxic respiration data resulting from a pulse addition of carbon source to endogenously respiring anoxic activated sludge shows a clear start-up phenomenon and storage tail that is usually also observed in high-frequency OUR measurements. Finally, the improved methodology can be expected to serve as an anoxic respirometer for activated sludge treatment plants where denitrification process occurs in single-step. Further, it can be used for a variety of purposes e.g. for toxicity and activity monitoring, process control and parameter estimation of the activated sludge process, similar to the aerobic respirometers.

Denitrifying activity measurements using an anoxic titration (pHstat) bioassay

An anoxic titrimetric test was investigated for measuring denitrifying activity in an activated sludge system. The method measures the amount of acid that is required to maintain the pH set-point value in a batch denitrification experiment. An iterative algorithm was implemented to extract nitrate uptake rate (NUR) data from titration data, since the accumulation and depletion (stripping) of reaction by-products HCO3- and CO2 affects the direct calculation of denitrifying activity from titration data. This method was performed using an automatic pH-stat acid dosing system, and the data were analysed using the simulation software package, AQUASIM.

Lupinus angustifolius has a plastic uptake response to heterogeneously supplied nitrate while Lupinus pilosus does not

Uptake rates calculated from plants uniformly supplied with a nutrient will underestimate uptake under heterogeneous conditions. A split-root nutrient solution experiment was set up to compare the uptake rate of 2 lupin species (Lupinus angustifolius L., L. pilosus Murr.) under conditions of uniform and heterogeneous nitrate supply. Nitrate was supplied uniformly to the root system at 250 M (low), 750 M (high), or 1500 M (high), or in a split low/high or high/low combination between the upper and lower root system. While L. pilosus had a greater total nitrate uptake over the treatment period due to a higher total root length, L. angustifolius had 1.5–2.5 times greater nitrate uptake rate per unit of root length. L. angustifolius also had the capacity to increase the nitrate uptake rate in sections of the root system supplied locally with high nitrate, compared with a root system uniformly supplied with high nitrate. This increased uptake rate under heterogeneous supply enabled the plant to take up 74–94% of the total nitrate taken up when uniformly supplied with high nitrate, while only 58–72% would have been taken up without such a compensation mechanism. L. pilosus did not show this response. The difference between the response of these 2 species suggests that a range of nitrate uptake responses may exist across the lupin germplasm, and that it may be possible to select a lupin species with an enhanced ability to capture nitrate from the profile, thus decreasing nitrate losses from leaching.