고등식물의 질산시그널에 의한 유전자 발현제어 관련 전사인자의 연구현황

정유진(Yu Jin Jung),박정순(Joung Soon Park),고지윤(Ji Yun Go),이효주(Hyo Ju Lee),김진영(Jin Young Kim),이예지(Ye Ji Lee),남기홍(Ki Hong Nam),조용구(Yong-Gu Cho),강권규(Kwon Kyoo Kang) 한국식물생명공학회 2021 JOURNAL OF PLANT BIOTECHNOLOGY Vol.48 No.3

Nitrate is an important nutrient and signaling molecule in plants that modulates the expression of many genes and regulates plant growth. In this study, we cover the research status of transcription factors related to the control of gene expression by nitrate signaling in higher plants. Nitrate reductase is a key enzyme in nitrogen assimilation, as it catalyzes the nitrate-to-nitrite reduction process in plants. A variety of factors, including nitrate, light, metabolites, phytohormones, low temperature, and drought, modulate the expression levels of nitrate reductase genes and nitrate reductase activity, which is consistent with the physiological role if. Recently, several transcription factors controlling the expression of nitrate reductase genes have been identified in higher plants. NODULE-INCEPTION-Like Proteins (NLPs) are transcription factors responsible for the nitrate-inducible expression of nitrate reductase genes. Since NLPs also control the nitrate-inducible expression of genes encoding the nitrate transporter, nitrite transporter, and nitrite reductase, the expression levels of nitrate reduction pathway-associated genes are coordinately modulated by NLPs in response to nitrate. Understanding the function of nitrate in plants will be useful to create crops with low nitrogen use.

Nitrate inhibits soybean nodulation by regulating expression of CLE genes

Lim, Chae Woo,Lee, Young Woo,Lee, Sung Chul,Hwang, Cheol Ho Elsevier 2014 Plant science Vol.229 No.-

<P><B>Abstract</B></P> <P>Nitrogen compounds such as nitrate act as a potential inhibitor for legume nodulation. In this study, we isolated a new CLE gene, <I>GmNIC2</I>, from nitrate-treated roots, which shares high sequence homology with nitrate-induced CLE gene <I>GmNIC1</I>. Similar to <I>GmNIC1</I>, the expression level of <I>GmNIC2</I> was not significantly altered in roots by rhizobial inoculation and was much higher in young nodules than in roots. In addition, overexpression of <I>GmNIC2</I> led to similar nodulation inhibition of transgenic hairy roots to that of <I>GmNIC1</I>, which occurred in GmNARK-dependent manner and at the local level. By analyzing GmNARK loss-of-function mutant, SS2-2, it was found that expression levels of <I>GmNIC1</I> and <I>GmNIC2</I> in the SS2-2 roots were lower than in the wild type (WT) roots in response to nitrate. In contrast to <I>GmNIC1</I> and <I>GmNIC2</I>, expressions of <I>GmRIC1</I> and <I>GmRIC2</I> genes that are related to the autoregulation of nodulation (AON) were strongly suppressed both of the soybeans during all periods of nitrate treatment and even were not induced by additional inoculation with rhizobia. Taken together, the results of this study suggest that GmNIC2, as an active homologous gene located in chromosome 13, acts locally to suppress nodulation, like GmNIC1, and nitrate inhibition of nodulation is led by fine-tuned regulation of both nitrate-induced CLEs and rhizobia-induced CLEs.</P> <P><B>Highlights</B></P> <P> <UL> <LI> A new nitrate-induced CLE gene, <I>GmNIC2</I>, is isolated from soybean roots. </LI> <LI> GmNIC2 acts as a local signal in nitrate inhibition of nodulation, like GmNIC1. </LI> <LI> Nitrate induces lower expression of <I>GmNIC</I> genes in the supernodulation mutant. </LI> <LI> Nitrate inhibits expression of rhizobia-induced <I>CLEs</I> after rhizobial inoculation. </LI> <LI> Nitrate inhibition of nodulation is led by a fine-tune regulation between CLEs. </LI> </UL> </P>

Effects of Nitrate Addition on Rumen Fermentation, Bacterial Biodiversity and Abundance

Liping Zhao,Qingxiang Meng,Liping Ren,Wei Liu,Xinzhuang Zhang,Yunlong Huo,Zhenming Zhou 아세아·태평양축산학회 2015 Animal Bioscience Vol.28 No.10

This study examined changes of rumen fermentation, ruminal bacteria biodiversity and abundance caused by nitrate addition with Ion Torrent sequencing and real-time polymerase chain reaction. Three rumen-fistulated steers were fed diets supplemented with 0%, 1%, and 2% nitrate (dry matter %) in succession. Nitrate supplementation linearly increased total volatile fatty acids and acetate concentration obviously (p = 0.02; p = 0.02; p<0.01), butyrate and isovalerate concentration numerically (p = 0.07). The alpha (p>0.05) and beta biodiversityof ruminal bacteria were not affected by nitrate. Nitrate increased typical efficient cellulolytic bacteria species (Ruminococcus flavefaciens, Ruminococcus ablus, and Fibrobacter succinogenes) (p<0.01; p = 0.06; p = 0.02). Ruminobactr, Sphaerochaeta, CF231, and BF311 genus were increased by 1% nitrate. Campylobacter fetus, Selenomonas ruminantium, and Mannheimia succiniciproducens were core nitrate reducing bacteria in steers and their abundance increased linearly along with nitrate addition level (p<0.01; p = 0.02; p = 0.04). Potential nitrate reducers in the rumen, Campylobacter genus and Cyanobacteria phyla were significantly increased by nitrate (p<0.01; p = 0.01).To the best of our knowledge, this was the first detailed view of changes in ruminal microbiota by nitrate. This finding would provide useful information on nitrate utilization and nitrate reducer exploration in the rumen.

Effect of Sodium Nitrate and Nitrate Reducing Bacteria on In vitro Methane Production and Fermentation with Buffalo Rumen Liquor

Sakthivel, Pillanatham Civalingam,Kamra, Devki Nandan,Agarwal, Neeta,Chaudhary, Chandra Asian Australasian Association of Animal Productio 2012 Animal Bioscience Vol.25 No.6

Nitrate can serve as a terminal electron acceptor in place of carbon dioxide and inhibit methane emission in the rumen and nitrate reducing bacteria might help enhance the reduction of nitrate/nitrite, which depends on the type of feed offered to animals. In this study the effects of three levels of sodium nitrate (0, 5, 10 mM) on fermentation of three diets varying in their wheat straw to concentrate ratio (700:300, low concentrate, LC; 500:500, medium concentrate, MC and 300:700, high concentrate, HC diet) were investigated in vitro using buffalo rumen liquor as inoculum. Nitrate reducing bacteria, isolated from the rumen of buffalo were tested as a probiotic to study if it could help in enhancing methane inhibition in vitro. Inclusion of sodium nitrate at 5 or 10 mM reduced (p<0.01) methane production (9.56, 7.93 vs. 21.76 ml/g DM; 12.20, 10.42 vs. 25.76 ml/g DM; 15.49, 12.33 vs. 26.86 ml/g DM) in LC, MC and HC diets, respectively. Inclusion of nitrate at both 5 and 10 mM also reduced (p<0.01) gas production in all the diets, but in vitro true digestibility (IVTD) of feed reduced (p<0.05) only in LC and MC diets. In the medium at 10 mM sodium nitrate level, there was 0.76 to 1.18 mM of residual nitrate and nitrite (p<0.01) also accumulated. In an attempt to eliminate residual nitrate and nitrite in the medium, the nitrate reducing bacteria were isolated from buffalo adapted to nitrate feeding and introduced individually (3 ml containing 1.2 to $2.3{\times}10^6$ cfu/ml) into in vitro incubations containing the MC diet with 10 mM sodium nitrate. Addition of live culture of NRBB 57 resulted in complete removal of nitrate and nitrite from the medium with a further reduction in methane and no effect on IVTD compared to the control treatments containing nitrate with autoclaved cultures or nitrate without any culture. The data revealed that nitrate reducing bacteria can be used as probiotic to prevent the accumulation of nitrite when sodium nitrate is used to reduce in vitro methane emissions.

사과나무의 질산염환원효소

이희재 서울대학교 농업개발연구소 2000 농업생명과학연구 Vol.4 No.-

optimize the assay conditions for various tissues of MM 106 apple trees. Optimum co Factors affecting the in vivo nitrate reductase assay were investigated in order to ncentrations of nitrate and phosphate for in vivo nitrate reductase activities ranged from 50 to 100 mM and were tissue-specific. The optimum pH of the assay medium was 7.5. Vacuum infiltration and the addition of 2% (v/v) n-propanol to the assay medium resulted in stimulation of nitrate reducatase activity. The in vivo nitrate reductase assays for leaves and stems were linear for at least 60 minutes following an initial 30 minute lag, whereas there was no lag phase in root tissues. The effect of nitrate supply in the nutrient solution on nitrogen accumulation and nitrate reductase activity was examined. Increasing nitrate supply resulted in increases in the concentration and amount of reduced nitrogen in leaves and stems. The reduced nitrogen contents of trunk bark and trunk wood were significantly decreased when low nitrate was supplied, whereas that of roots was changed insignificantly. Most organs of MM 106 apple trees contained only a small portion of their total nitrogen as nitrate. The activity of nitrate reductase was measured using in vivo assay either with or without nitrate, namely in vivo+NO3 and in vivo- NO3 nitrate reductase assay. The in vivo- NO3 assay may be the better way to estimate reduced nitrogen accumulation than the in vivo+ NO3 assay. Increasing nitrate supply resulted in significant increases of in vivo- NO3 nitrate reductase activities of all organs. The proportion of leaf nitrate reductase activity to the total activity increased as the nitrate supply increased. The stems also contributed considerably to the total nitrate reducatase activity. Therefore, it can be concluded that nitrate reduction can occur in various parts of MM 106 apple trees as long as nitrate is available. Effect of darkness on intrate reduction was also examined in MM 106 apple leaves. As the period of darkness increased, nitrate reductase activity decreased and the accumulation of nitrate increased. The decrease in nitrate reductase activity with increasing period of darkness may be related to the decrease in soluble carbohydrates.

Nitrate Uptake in the Halotolerant Cyanobacterium Aphanothece halophytica is energy-dependent driven by ΔpH

Incharoensakdi, Aran,Laloknam, Surasak Korean Society for Biochemistry and Molecular Biol 2005 Journal of biochemistry and molecular biology Vol.38 No.4

The energetics of nitrate uptake by intact cells of the halotolerant cyanobacterium Aphanothece halophytica were investigated. Nitrate uptake was inhibited by various protonophores suggesting the coupling of nitrate uptake to the proton motive force. An artificially-generated pH gradient across the membrane (${\Delta}pH$) caused an increase of nitrate uptake. In contrast, the suppression of ${\Delta}pH$ resulted in a decrease of nitrate uptake. The increase of external pH also resulted in an enhancement of nitrate uptake. The generation of the electrical potential across the membrane ($\Delta\psi$) resulted in no elevation of the rate of nitrate uptake. On the other hand, the valinomycin-mediated dissipation of $\Delta\psi$ caused no depression of the rate of nitrate uptake. Thus, it is unlikely that $\Delta\psi$ participated in the energization of the uptake of nitrate. However, $Na^+$-gradient across the membrane was suggested to play a role in nitrate uptake since monensin which collapses $Na^+$-gradient strongly inhibited nitrate uptake. Exogenously added glucose and lactate stimulated nitrate uptake in the starved cells. N, N'-dicyclohexylcarbodiimide, an inhibitor of ATPase, could also inhibit nitrate uptake suggesting that ATP hydrolysis was required for nitrate uptake. All these results indicate that nitrate uptake in A. halophytica is ATP-dependent, driven by ${\Delta}pH$ and $Na^+$-gradient.

Effects of Nitrate Addition on Rumen Fermentation, Bacterial Biodiversity and Abundance

Zhao, Liping,Meng, Qingxiang,Ren, Liping,Liu, Wei,Zhang, Xinzhuang,Huo, Yunlong,Zhou, Zhenming Asian Australasian Association of Animal Productio 2015 Animal Bioscience Vol.28 No.10

This study examined changes of rumen fermentation, ruminal bacteria biodiversity and abundance caused by nitrate addition with Ion Torrent sequencing and real-time polymerase chain reaction. Three rumen-fistulated steers were fed diets supplemented with 0%, 1%, and 2% nitrate (dry matter %) in succession. Nitrate supplementation linearly increased total volatile fatty acids and acetate concentration obviously (p = 0.02; p = 0.02; p<0.01), butyrate and isovalerate concentration numerically (p = 0.07). The alpha (p>0.05) and beta biodiversityof ruminal bacteria were not affected by nitrate. Nitrate increased typical efficient cellulolytic bacteria species (Ruminococcus flavefaciens, Ruminococcus ablus, and Fibrobacter succinogenes) (p<0.01; p = 0.06; p = 0.02). Ruminobactr, Sphaerochaeta, CF231, and BF311 genus were increased by 1% nitrate. Campylobacter fetus, Selenomonas ruminantium, and Mannheimia succiniciproducens were core nitrate reducing bacteria in steers and their abundance increased linearly along with nitrate addition level (p<0.01; p = 0.02; p = 0.04). Potential nitrate reducers in the rumen, Campylobacter genus and Cyanobacteria phyla were significantly increased by nitrate (p<0.01; p = 0.01).To the best of our knowledge, this was the first detailed view of changes in ruminal microbiota by nitrate. This finding would provide useful information on nitrate utilization and nitrate reducer exploration in the rumen.

Nitrate supplementation of rations based on rice straw but not Pangola hay, improves growth performance in meat goats

Paengkoum Siwaporn,Khotsakdee Jiravan,Paengkoum Pramote,Schonewille Thomas,Yuangklang Chalermpon 아세아·태평양축산학회 2021 Animal Bioscience Vol.34 No.6

Objective: Supplemental nitrate is known to be an effective tool to mitigate methane emission by ruminants. Based on theoretical considerations, supplemental nitrate can improve but also deteriorate the growth performance. The overall effect of supplemental nitrate on growth performance, however, is not yet known. The objective of the current study was therefore to evaluate the effect of a higher dose of NO3 – on overall growth performance when feeding either Pangola grass hay or rice straw. Methods: Thirty-two crossbred, 3-month-old Thai native×Anglo-Nubian crossbred male goats were used. The experiment had a 2×2 factorial design with an experimental period of 60 days. Eight goats were randomly allocated to each dietary treatment, i.e. a ration containing either Pangola hay (Digitaria eriantha Steud) or rice straw (Oryza Sativa) as a source of roughage, supplemented with a concentrate containing either 3.2% or 4.8% potassium nitrate. The rations were formulated to be isonitrogenous. The animals were weighed at the start of the experiment and at days 30 and 60. Feces were collected during the last five days of each 30-day period. Results: High-nitrate increased overall DM intake by approximately 3%, irrespective the source of roughage, but only the goats fed a rice straw-based ration responded with an increase in body weight (BW). Thus, the overall feed conversion ratio (kg feed/kg BW gain) was influenced by roughage source ×nitrate and decreased by almost 60% when the goats were fed rice straw in combination with a high versus a low dietary nitrate content. The digestibility of macronutrients was only affected by the source of roughage and the digestibility of organic matter, crude protein, and neutral detergent fibre was greater when the goats were fed Pangola hay. Conclusion: It was concluded that the replacement of soybean meal by nitrate improves the growth performance of meat goats fed rations based on rice straw, but not Pangola hay. Objective: Supplemental nitrate is known to be an effective tool to mitigate methane emission by ruminants. Based on theoretical considerations, supplemental nitrate can improve but also deteriorate the growth performance. The overall effect of supplemental nitrate on growth performance, however, is not yet known. The objective of the current study was therefore to evaluate the effect of a higher dose of NO₃^– on overall growth performance when feeding either Pangola grass hay or rice straw.Methods: Thirty-two crossbred, 3-month-old Thai native×Anglo-Nubian crossbred male goats were used. The experiment had a 2×2 factorial design with an experimental period of 60 days. Eight goats were randomly allocated to each dietary treatment, i.e. a ration containing either Pangola hay (<i>Digitaria eriantha Steud</i>) or rice straw (<i>Oryza Sativa</i>) as a source of roughage, supplemented with a concentrate containing either 3.2% or 4.8% potassium nitrate. The rations were formulated to be isonitrogenous. The animals were weighed at the start of the experiment and at days 30 and 60. Feces were collected during the last five days of each 30-day period.Results: High-nitrate increased overall DM intake by approximately 3%, irrespective the source of roughage, but only the goats fed a rice straw-based ration responded with an increase in body weight (BW). Thus, the overall feed conversion ratio (kg feed/kg BW gain) was influenced by roughage source ×nitrate and decreased by almost 60% when the goats were fed rice straw in combination with a high versus a low dietary nitrate content. The digestibility of macronutrients was only affected by the source of roughage and the digestibility of organic matter, crude protein, and neutral detergent fibre was greater when the goats were fed Pangola hay.Conclusion: It was concluded that the replacement of soybean meal by nitrate improves the growth performance of meat goats fed rations based on rice straw, but not Pangola hay.

염기성 pH에서 Enterobacter amnigenus GG0461의 질산이온 흡수증가

최태근,김성태,한민우,김영기 한국응용생명화학회 2008 Journal of Applied Biological Chemistry (J. Appl. Vol.51 No.1

Salt accumulation in soils of greenhouse due to the massive application of nitrogen fertilizers causes salt stress on the various crops, a serious problem in domestic agriculture. Since the majority of the salinity is nitrate, the excess nitrate should be removed; therefore, a bacterial strain having high capacity of nitrate uptake and identified as Enterobacter amnigenus GG0461 was isolated from the soils of greenhouse. Optimum conditions for the bacterial growth and nitrate uptake were investigated. GG0461 was able to grow without nitrate; however, nitrate facilitated the growth. The rate of nitrate uptake increased at alkaline pH and both growth and nitrate uptake were maximal at pH 8-9. When the initial pH of culture medium was increased to pH 8 or 9, it was decreased to neutral upon bacterial growth and nitrate uptake. These results imply that the major factor mediating bacterial nitrate uptake is a nitrate/proton antiporter. The fact was supported by the effect of nitrate addition in the absence of nitrate, since the addition of nitrate greatly increased the nitrate uptake and rapidly decreased pH of media. 질소비료의 과량 시용에 따른 시설원예지 토양 중 염류집적은 다양한 작물에 염류장애를 유발하여 국내농업에 심각한 문제가 되고 있다. 염류의 주성분은 질산염으로 필요이상의 질산 이온을 제거하기 위하여 질산이온 흡수력이 크며, Enterobacter amnigenus GG0461로 동정된 토양세균을 시설원예 토양에서 분리하였다. 이 균주의 최적 생육 및 질산이온 흡수력을 조사하였을 때, GG0461 균주는 질산이온이 없는 조건에서 생육이 가능하나, 질산이온이 존재할 때 생육이 크게 촉진되었다. 또한, 염기성 조건에서 질산이온 흡수율이 증가하였으며, 성장과 질산이온 흡수 모두 pH 8-9에서 최대로 나타났다. 배지의 초기 pH를 8과 9로 조정하였을 때, 균주에 의하여 질산이온이 흡수 됨에 따라 배지의 pH는 중성으로 감소하였다. 이러한 결과는 질산이온 흡수를 위한 주된 인자가 nitrate/proton antiporter임을 의미한다. 이것은 질산이온이 없는 조건에서 질산이온의 첨가 효과인 질산이온 흡수의 급격한 증가와 빠른 배지의 pH 감소 사실로도 확인되었다.

Effects of Combination of Nitrate with ${\beta}$1-4 Galacto-oligosaccharides and Yeast (Candida kefyr) on Methane Emission from Sheep

Sar, C.,Santoso, B.,Gamo, Y.,Kobayashi, T.,Shiozaki, S.,Kimura, K.,Mizukoshi, H.,Arai, I.,Takahashi, J. Asian Australasian Association of Animal Productio 2004 Animal Bioscience Vol.17 No.1

The objective of the present study was to determine whether ${\beta}$1-4 galacto-oligosaccharides (GOS) and Candida kefyr combined with nitrate as manipulators could suppress rumen methanogenesis without nitrate poisoning in sheep. Four rumen fistulated wethers were allocated to a $4{\times}4$ Latin square design. Nitrate (1.3 g $NaNO_3$ $Kg^{-0.75}$body weight) with and without GOS and Candida kefyr were administered into the rumen through fistula as a single dose 30 min after the morning meal. GOS and Candida kefyr were supplemented by sprinkling onto the feed and through rumen fistula, respectively. The four treatments consisted of saline, nitrate, nitrate plus GOS and nitrate plus GOS plus Candida kefyr. Physiological saline was used as the control treatment. Compared to saline treatment, the administration of nitrate alone resulted in a very marked decrease in rumen methanogenesis and an increase in rumen and plasma nitrite production and blood methaemoglobin formation consequently causing a decline in oxygen consumption, carbon dioxide production and metabolic rate. When compared to nitrate alone, the simultaneous administration of nitrate with GOS decreased nitrite accumulation in rumen and plasma and nitrate-induced methaemoglobin, while retaining low methane production. However, GOS could not fully restore metabolic parameters reduced by nitrate. When compared to the simultaneous administration of nitrate with GOS, the simultaneous administration of nitrate with GOS plus Candida kefyr lowered rumen methanogenesis to a negligible level, but did not decrease rumen and plasma nitrite accumulation as well as blood methaemoglobin formation. Thus, these results suggest that combination of nitrate with GOS may be a potent manipulator to suppress rumen methanogenesis with abating the hazards of nitratenitrite toxicity in ruminants.