• <tr id="yyy80"></tr>
  • <sup id="yyy80"></sup>
  • <tfoot id="yyy80"><noscript id="yyy80"></noscript></tfoot>
  • 99热精品在线国产_美女午夜性视频免费_国产精品国产高清国产av_av欧美777_自拍偷自拍亚洲精品老妇_亚洲熟女精品中文字幕_www日本黄色视频网_国产精品野战在线观看 ?

    Brassinosteroids modulate nitrogen physiological response and promote nitrogen uptake in maize (Zea mays L.)

    2022-02-19 09:31:38JipengXingYubinWngQingqingYoYushiZhngMingciZhngZhohuLi
    The Crop Journal 2022年1期

    Jipeng Xing, Yubin Wng,b, Qingqing Yo, Yushi Zhng, Mingci Zhng,*, Zhohu Li

    a State Key Laboratory of Plant Physiology and Biochemistry, College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China

    b Crop Research Institute, Shandong Academy of Agricultural Sciences, Jinan 250100, Shandong, China

    Keywords:Brassinosteroids Nitrogen uptake Nitrate transporter gene Root architecture Maize

    ABSTRACT Brassinosteroids(BRs)are steroid hormones that function in plant growth and development and response to environmental stresses and nutrient supplies.However,few studies have investigated the effect of BRs in modulating the physiological response to nitrogen(N)supply in maize.In the present study,BR signalingdeficient mutant zmbri1-RNAi lines and exogenous application of 2,4-epibrassinolide (eBL) were used to study the role of BRs in the regulation of physiological response in maize seedlings supplied with N.Exogenous application of eBL increased primary root length and plant biomass,but zmbri1 plants showed shorter primary roots and less plant biomass than wild-type plants under low N(LN)and normal N(NN)conditions.LN induced the expression of the BR signaling-associated genes ZmDWF4, ZmCPD, ZmDET2,and ZmBZR1 and the production of longer primary roots than NN.Knockdown of ZmBRI1 weakened the biological effects of LN-induced primary root elongation.eBL treatment increased N accumulation in shoots and roots of maize seedlings exposed to LN or NN treatment.Correspondingly,zmbri1 plants showed lower N accumulation in shoots and roots than wild-type plants.Along with reduced N accumulation, zmbri1 plants showed lower fluxes and 15 uptake.The expression of nitrate transporter (NRT) genes(ZmNPF6.4, ZmNPF6.6, ZmNRT2.1, ZmNRT2.2) was lower in zmbri1 than in wild-type roots, but eBL treatments up-regulated the transcript expression of NRT genes.Thus,BRs modulated N physiological response and regulated the transcript expression of NRT genes to promote N uptake in maize.

    1.Introduction

    Brassinosteroids (BRs) are plant-specific steroidal hormones involved in plant growth and developmental processes and response to various abiotic stresses.Studies have revealed BRs influence root architecture, which determinies the efficiency of water and nutrient acquisition in higher plants.High concentrations of BRs inhibit, and low concentrations promote, root growth[1,2].InArabidopsis,the BR-insensitive mutantsbri1-116andbes1-D(gain-of-function mutants) exhibit short roots [3,4].The root elongation ofzmbri1-RNAi plants is insensitive to brassinolide treatment [5].Root development is a highly plastic process that is sensitive to nutrient limitation and other environmental parameters, with plant hormones acting as signaling mechanisms [6,7].BRs are involved in nitrogen (N)-modulated root elongation processes inArabidopsis.In a whole-genome association analysis,BRASSINOSTEROID SIGNALING KINASE3 (BSK3) functioned in root elongation response under low N condition[8].The BR biosynthesis geneAtDWF1is also a key gene involved in the root elongation response induced by N deficiency,and its overexpression increased N accumulation [9].These findings support the role of BRs in root development modulated by nutrition absorption.However, the function of BRs in N-modulated root architecture development in crops, especially monocots, has been little studied.

    Recent research has focused on the role of BRs in plant growth and development adapted to nutrient-deficiency stress [10].In rice, iron (Fe) deficiency inhibits BR biosynthesis, and reduced endogenous BRs contribute to Fe transport and translocation from roots to shoots [11].Besides their involvement in root development in response to N deficiency,recent studies [12,13] have suggested that BRs are also involved in N uptake and metabolism processes.BRs regulate ammonium () uptake via regulation oftransporters inArabidopsisand rice.BR signaling acts in tomato plant response to N starvation by regulating autophagy[14].In cucumber, exogenous application of 2,4-epibrassinolide(eBL)reduced the harmful effects of suboptimal root zone temperature by changing the N metabolism and the flux rates ofand nitrate (), thereby increasing the growth of seedlings [15].These studies suggest positive roles of BRs in plant N uptake.

    Genetic and breeding studies in maize,a major food and industrial crop, have shown that BRs influence plant architecture and kernel size regulation,thereby contributing to increase grain yield.Maize BR-deficient mutants have shortened internodes, twisted,dark green, erect leaves, and feminized male flowers [5,31,32].

    ZmRAVL1regulatesbrd1(brassinosteroid C-6 oxidase1) to alter endogenous BR content.ZmRAVL1-edited lines displayed reduced leaf angle and compacted plant architecture leading to increased plant density and potential yield [33].ZmBES1/BZR1-5 positively regulates maize kernel size [34].

    In maize production,external N inputs and their soil availability are the main nutrient-limiting factors in yield,and increasing N use efficiency is a goal of production[35].Identifying the physiological and molecular mechanisms of N uptake and utilization would further the breeding of N-efficient maize cultivars.N supply mediated the biosynthesis of BRs to modulate root elongation inArabidopsis[9].Overexpression ofZmDWF4increased grain yield by enhancing photosynthetic ability,promoting the expression of genes regulating cell division and the storage reservoir in maize kernels [36].Exogenous application of BR regulated maize growth and biomass accumulation to increase yield under either stress or normal field condition [37,38].The balance between carbon and N is the basis of high yield in crop production, but little is known about how BRs modulateuptake in maize plants in response to N supply.

    The aim of this study was to investigate the role of BRs in the regulation of physiological responses to N supply in maize.The BR signaling-deficient mutantzmbri1-RNAi and exogenous eBL treatment were used for measuring the N uptake under low N(LN) and normal N (NN) conditions.A noninvasive micro-test(NMT) and15N labeling were used for evaluating BR-mediateduptake.

    2.Materials and methods

    2.1.Plant materials and growth conditions

    ZmBRI1RNA interference(RNAi)plants were obtained from the Genetics,Development,and Cell Biology Department of Iowa State University [5].The gene expression ofZmBRI1was decreased inzmbri1plants (Fig.S1).Seeds of wild-type (WT) maize (Zea maysL.cv.B73) andzmbri1plants were surface sterilized in a 10% (v/v) H2O2solution for 20 min and germinated on sand for 7 days in a growth chamber, at 28/22 °C with a 16 h/8 h light/dark cycle and 70%-80%relative humidity.Uniform seedlings with two visible leaves were transferred to half-strength culture solution for 2 days and then to full-strength solution (0.5 mmol L-1MgSO4,0.1 mmol L-1KH2PO4, 1 mmol L-1CaCl2, 0.1 mmol L-1EDTA-Fe,0.03 mmol L-1H3BO3, 0.0025 mmol L-1ZnSO4, 0.008 mmol L-1CuSO4, 0.005 mmol L-1MnSO4,and 0.0003 mmol L-1(NH4)6Mo7-O24),but supplied with differentconcentrations(2.0 mmol L-1KNO3,NN;0.05 mmol L-1KNO3,LN).The concentration of K+in the LN solution was supplemented with KCl to the same level as that in the NN solution.The nutrient solution was renewed every 3 days.When seedlings were transferred either the NN or the LN solution,treatment with 2,4-epibrassinolide (eBL) (RealTimes, CAS:78821-43-9) was applied simultaneously.The eBL was first dissolved in ethanol to make a stock liquor and then added into nutrient solution to a final concentration of 0.05 nmol L-1at every change of solution.The same dose of ethanol was added to the control.

    2.2.Measurement of root length and plant biomass

    Uniform seedlings from each treatment were harvested and separated into roots and shoots 6 days after NN and LN treatments.Primary root length was measured with a ruler.The relative rate of primary root length induced by LN is calculated by (LN - NN)/LN.Total root length and surface area were determined with WinRHIZO software (Pro 2014b, Beijing, China) from images acquired with an Epson V700 (Seiko Epson Corp, Nagano-ken, Japan) scanner.Samples for measuring plant biomass were heated for 30 min at 105 °C and dried for 3 days at 75 °C and dry weight was recorded using a electronic balance.Ten biological replicates were used for each treatment.

    2.3.Measurement of fluxes at the root surface by NMT

    2.4.15N uptake measurement and estimation of total N content

    After LN or NN treatment for 5 days, uniform seedlings were selected for15N labeling.Roots were rinsed with 0.1 mmol L-1CaSO4solution for 1 min and transferred into nutrient solution with 0.05 or 2 mmol L-1K15NO3with a 99% atom excess of15N for 10 min.They were then rinsed for 1 min in 0.1 mmol L-1CaSO4solution before sampling.Samples were harvested separately and oven-dried for 3 days at 75 °C to constant weight.Dry samples were weighed and ground.The powder was used for total15N determination by isotope ratio mass spectrometry (Vario PYRO cube ISOprime 100, Isoprime/Elementar Ltd., Cheadle Hulme,UK).Total N accumulation in roots and shoots was estimated following Bremner et al.[40].Plant total N accumulation was calculated as the product of N concentration and corresponding dry weight.Three biological samples were used for each treatment.

    2.5.RNA extraction and quantitative RT-PCR (qRT-PCR) analysis

    Total RNA was isolated from each sample using an EASYspin rapid plant extraction kit(Aialab,Beijing,China),and reverse transcription was preformed using Oligo d (T) primer and M-MLV reverse transcriptase (Takara, Japan).The qRT-PCR was conducted in an Applied Biosystems 7500 Fast Real-Time PCR System(Applied Biosystems, USA) using TB Green Premix Ex Taq II (Takara, Japan).Gene expression was calibrated to the expression ofZmUBC(ubiquitin C).The relative gene expression was calculated by the 2-ΔΔCTmethod [41].Three biological replicates were used for each treatment.The primers for qRT-PCR are listed in Table S1.

    2.6.Statistical analysis

    Analysis of variance was performed using the general linear model (GLM) procedure in SPSS 21.0 (SPSS Inc., Chicago, IL, USA).Means of more than two groups were compared by Duncan’s multiple-range test atP<0.05.Changes in biomass and N accumulation were tested with Fisher’s LSD test atP< 0.05.

    3.Results

    3.1.BRs increased plant growth in response to N supply

    As shown in Fig.1a,LN inhibited the growth of roots and shoots compared to NN, while eBL treatment promoted shoot and root growth under both N conditions.The relative rate of primary root length induced by LN was 9.6% in eBL-treated plants and 7.3% in control plants(Fig.1b).In comparison with control plants,the total root length and root surface area increased by respectively 17.4%and 16.4% in eBL-treated plants under the NN condition and by 22.9% and 20.2% under the LN condition (Fig.S2).Shoot and root biomass increased by respectively 21.0% and 15.0% in eBL-treated plants under the NN condition and by 24.6% and 19.9% under the LN condition (Fig.1c, d).However, the ratio of root to shoot showed no significant difference between eBL-treated and control plants under LN and NN conditions (Fig.1e).

    Thezmbri1plants showed a dwarf phenotype, lower primary root length, total root length, and root surface area than wildtype plants under both N conditions (Fig.2a).The relative rate of primary root length induced by LN was 1.2%-2.3%inzmbri1plants but 9.1% in wild-type plants (Fig.2b).In comparison with wildtype plants, total root length and root surface area decreased by respectively 50.6%-51.8% and 58.2%-60.8% inzmbri1plants under the NN condition but by 52.3%-52.5% and 59.4%-61.5% under the LN condition (Fig.S3).Shoot and root biomass decreased by 61.2%-63.8%and 54.2%-60.7%inzmbri1plants in comparison with wild-type plants under the NN condition, but by 62.1%-64.3% and 53.6%-61.2%under the LN condition(Fig.2c,d).The ratio of root to shoot was higher inzmbri1plants than in wild-type plants(Fig.2e).

    3.2.N supply was involved in mediating the transcript expression of BR biosynthetic and signaling genes

    As shown in Fig.3a, LN significantly induced the expression ofZmDWF4compared to NN at 1-6 days after N treatments.The expression levels ofZmCPDandZmDET2in LN-treated seedlings were also higher than those in NN-treated plants (Fig.3b, c).LN significantly increased the expression levels ofZmBZR1, a gene mediating pleiotropic BR responses, in comparison with NN at 1-6 days after N treatments (Fig.3d).

    3.3.BRs influenced N accumulation in maize seedlings subjected to N supply

    To determine the role of BRs involved in N uptake in maize seedlings, N accumulation was measured in eBL-treated plants and zmbri1plants under both N conditions.Under NN condition,eBL treatment significantly increased N accumulation in shoots and roots in comparison with control plants.N accumulation in shoots and roots in eBL-treated plants increased by respectively 11.6%and 31.5%under the LN condition(Fig.4a).N accumulations in shoots and roots inzmbri1plants were lower than those in wildtype plants under LN and NN conditions.They decreased by 49.8%-51.9% and 43.9%-48.1% under NN and by 57.3%-57.8% and 50.1%-53.1% under LN (Fig.4b).

    To test whether the effects of BRs on N accumulation were due to plant growth or nutritional status,the dynamic accumulation of dry biomass and total N accumulation were measured in wild-type andzmbri1plants under LN and NN conditions.LN reduced the dry biomass of shoot and root compared to NN, whilezmbri1plants presented lower dry biomass of shoot and root than wild-type plants at 1-9 days after LN or NN treatment (Fig.5a, b).Although the dry biomass of shoot and root was lower inzmbri1plants than in wild-type plants under both LN and NN conditions,there was no significant difference between wild-type andzmbri1plants in the relative inhibition rate by LN to NN of dry biomass(Fig.5c,d).Similarly,zmbri1plants showed lower N accumulation in shoots and roots than wild-type plants at 1-9 days after LN or NN treatment(Fig.5e, f).However,zmbri1plants showed higher relative inhibition rates of LN in N accumulation in shoots and roots than wildtype plants at 1-9 days after N treatments (Fig.5g, h).

    3.4.BRs were involved in modulating NO3- uptake

    To clarify the function of BRs in modulating N uptake, netfluxes were measured by NMT in wild-type andzmbri1roots.As shown in Fig.6a, LN significantly repressed netfluxes in wild-type andzmbri1roots in comparison with NN, while netfluxes were markedly decreased inzmbri1roots relative to those in wild-type roots under LN and NN conditions.In comparison with wild-type roots, meanfluxes were decreased by 31.7%-45.9% inzmbri1roots under the NN condition and by 37.1%-57.5% inzmbri1roots under the LN condition (Fig.6b).The15N tracing assay suggested that the amount of15N inzmbri1plants was less than that in wild-type plants, while LN reduced the amount of15N in wild-type andzmbri1plants (Fig.6c).15fluxes also decreased inzmbri1plants (Fig.6d).

    3.5.BRs regulated the transcript expression of NO3- uptake-associated genes

    To investigate how BRs altered N uptake, the expression levels ofuptake-related genes were measured in eBL-treated,wild-type, andzmbri1roots responding to NN and LN.As shown in Fig.7a, the transcription levels ofZmNPF6.4were significantly decreased inzmbri1roots in comparison with wild-type roots,while LN down-regulated the expression ofZmNPF6.4in wildtype andzmbri1roots at 6 days after N treatments.Similarly toZmNPF6.4, the expression levels ofZmNPF6.6were lower inzmbri1roots than those in wild-type roots under both LN and NN conditions at 1-6 days after N supply (Fig.7b).Compared to NN, the expressions ofZmNRT2.1andZmNRT2.2were down-regulated at 3 and 6 days after LN treatment in wild-type andzmbri1roots(Fig.7c-d).The expression levels of these genes inzmbri1roots were lower than those in wild-type roots under LN and NN conditions.

    Fig.1.Changes in morphology and plant biomass in control and eBL-treated plants in response to N supply.(a)Phenotypic characteristics of eBL-treated plants at 6 days after LN or NN.Scale bars,10 cm.(b-e)Effects of eBL on primary root length(b),shoot dry weight(c),root dry weight(d),and ratio of root to shoot(e)in maize seedlings under LN and NN conditions.The control was maize cv.B73.Values with error bars represent mean±SD(n=10).Different letters indicate significant difference between treatments according to Duncan’s multiple range test at P < 0.05.eBL, 2,4-epibrassinolide; LN, low N; NN, normal N.

    Exogenous eBL application was used to further clarify the regulation by BRs of the transcript expression ofuptake-associated genes under both LN and NN conditions.eBL treatment significantly up-regulated the expression ofZmNPF6.4andZmNPF6.6relative to the control at 1-6 days after N supply (Fig.7e, f).eBLtreated plants showed higher transcript expression levels ofZmNRT2.1andZmNRT2.2than control plants under both LN and NN conditions (Fig.7g, h).

    4.Discussion

    Brassinosteroids are growth-promoting steroidal hormones that play important roles in physiological and developmental adaptation to plants environment conditions [42-44].BRs are also involved in modulating the adaptation of plants in response to the supply of nutrients such as Fe and Pi [11,45].Several studies[10] have suggested that BRs play important roles in mediating N uptake and metabolism processes in plants.In the present study,exogenous eBL treatment and the BR signaling-deficient mutantzmbri1-RNAi affected the accumulation of shoot and root biomass,and the ratio of root to shoot in maize seedlings in response to LN or NN.These findings suggested that BR accumulation affects the adaptation of the maize plant to N supply in the soil.LN induced the expression of the BR signaling-associated genesZmDWF4,ZmCPD,ZmDET2,andZmBZR1and the production of longer primary roots than NN.Similarly,low N regulated the biosynthesis of BRs to stimulate root elongation inArabidopsis[9].Thus, BRs influenced the response to N supply in maize.

    Fig.2.Changes in morphology and plant biomass in zmbri1 plants under LN and NN conditions.(a)Phenotypic characteristics of zmbri1 plants at 6 days after LN or NN.Scale bars, 10 cm.(b-e) Changes in primary root length (b), shoot dry weight (c), root dry weight (d), and ratio of root to shoot (e) in zmbri1 plants under LN and NN conditions.Values with error bars are mean±SD(n=10).Different letters indicate significant difference between treatments by Duncan’s multiple range test at P<0.05.WT,wild type;LN, low N; NN, normal N.

    Fig.3.Effects of N supply on expression of ZmDWF4 (a),ZmCPD (b),ZmDET2(c), and ZmBZR1 (d) in maize (cv.B73)seedlings.Values with error bars are mean±SD(n=3).Different letters indicate significant difference between treatments by Duncan’s multiple range test at P < 0.05.LN, low N; NN, normal N.

    Fig.4.Effects of BRs on N accumulation under different N conditions.(a)N accumulation in shoots and roots of maize seedlings supplemented or not with eBL at 6 days after LN or NN supply.(b)N accumulation in shoots and roots of wild-type and zmbri1 plants at 6 days after LN or NN supply.The control and WT are maize cv.B73.Values with error bar are mean±SD(n=3).Different letters indicate significant difference between treatments in shoots and roots by Duncan’s multiple-range test at P<0.05.eBL,2,4-epibrassinolide; WT, wild type; LN, low N; NN, normal N.

    Root architecture plasticity is crucial for many plant species suffering from soil nutrient limitations, and N supply affects root growth and development inArabidopsis, rice, and maize [46-48].Studies [49,50] suggest that BRs influence the root growth and development of various plant species.BR-deficient or -signaling mutants displayed short-root phenotypes [1,2,51].Exogenous low concentrations of BRs increased root growth, whereas high concentrations of BRs repressed it [1,52].In the present study,exogenous application of eBL promoted primary root growth under both LN and NN conditions, whereaszmbri1plants showed shortroot phenotypes in contrast to wild-type plants.LN induced growth of primary roots in maize seedlings, and exogenous eBL promoted the effects of LN-mediated primary root elongation,whereas knockdown ofZmBRI1weakened the biological effects of LN-induced primary root elongation.In a previous study [8], LN up-regulated the transcript of the BR co-receptor BAK1 to activate BR signaling and stimulate root elongation, whereas a mutant ofbsk3in BR signaling effectively repressed low N-mediated primary root elongation.LN also up-regulated the expression of BR biosynthesis geneAtDWF1contributed to the root foraging response [9].In agreement with these results, the present study suggested that BRs were involved in-modulated root architecture in maize seedlings.Further research could reveal the physiological and molecular mechanism by which BRs influence-modulatedroot elongation.

    Fig.5.Changes in plant biomass and N accumulation in wild-type and zmbri1 plants under LN and NN conditions.(a,b)Dynamics changes in shoot(a)and root(b)dry weight of wild-type and zmbri1 plants at 1,3,6,and 9 days after LN or NN supply.Vertical bars indicate LSD at 0.05 levels.(c,d)Relative inhibition rate in shoot(c)and root(d)dry weight of wild-type and zmbri1 plants.Relative inhibition rate = (NN - LN)/NN.(e, f) Dynamic changes in N accumulation in shoot (e) and root (f) of wild-type and zmbri1 plants at 1,3,6,and 9 days after LN or NN supply.Vertical bars indicate LSD at 0.05 levels.(g,h).Relative inhibition rate of N accumulation in shoot(g)and root(h)of wildtype and zmbri1 plants.Values with error bars are mean±SD(n=3).Different letters indicate significant difference between different treatments at the same time point by Duncan’s multiple-range test at P < 0.05.WT, wild type; LN, low N; NN, normal N.

    Fig.6.Knockdown of ZmBRI1 influenced uptake.(a,b)Net fluxes(a)and mean fluxes(b)in roots of wild-type and zmbri1 plants under LN and NN conditions.Values with error bars are mean±SD(n=8).(c,d) 15N content(c)and 15influxes(d)after 10 min 15N tracing assay in wild-type and zmbri1 roots.Values with error bars are mean±SD(n=3).Different letters indicate significant difference between treatments by Duncan’s multiple range test at P<0.05.WT,wild type;LN,low N;NN,normal N.

    BRs function in the regulation of physiological and developmental processes in plants and also modulate N uptake and metabolism in plants [13,53].In the present study, exogenous application of eBL increased N accumulation in shoots and roots of maize seedlings exposed to LN or NN treatment.Correspondingly,zmbri1plants showed lower N accumulation in shoots and roots than did wild-type plants under both LN and NN conditions.Knockdown ofZmBRI1altered the relative inhibition rate by LN of N accumulation.Although the lower growth rate of zmbri1plants led to low nutrient demand, the relative inhibition rate of dry biomass in shoots showed no significant difference between wild-type andzmbri1plants under the LN condition, while that inzmbri1plants showed a slight upward tendency in comparison with wild-type plants subjected to extended LN treatment.Thus, the higher inhibition rate of N accumulation inzmbri1plants appears not to be the major limiting factor in plant biomass accumulation during short-term N deficiency, and thezmbri1plant biomass inhibition rate may depend on the cumulative time under LN conditions.

    Associated with their N accumulation,zmbri1plants showed lowerfluxes and15uptake than wild-type plants under both LN and NN conditions.Similar results were observed [15] in cucumber, where exogenous application of eBL promotedandflux rates.These findings indicate that BRs are involved in the process of N uptake for increasing N accumulation in maize seedlings.NRT protein families have been shown [20] to be involved inabsorption from soil and translocation to various plant tissues in many species.The expression of NRT genes is regulated by auxin, ethylene, cytokinin, and gibberellin in plants in response to N supply [30,54].Previous studies [15,53] indicated that BRs regulated the transcription of NPF/NTR1 genes inArabidopsisand cucumber.ZmNPF6.6is a dual-affinity nitrate transporter protein, whereasZmNPF6.4has low-affinitytransport activity [22].In the present study, the expressions ofZmNPF6.4andZmNPF6.6were down-regulated inzmbri1roots in comparison with wild-type roots under LN and NN conditions.In contrast,eBL treatment increased the transcript expression ofZmNPF6.4andZmNPF6.6relative to the control under both LN and NN conditions.AtNRT2.1is the major high-affinity transport gene responsible foruptake at low concentrations ofinArabidopsis[17].The expression levels ofZmNRT2.1andZmNRT2.2,two maize homologs ofAtNRT2.1were also correlated withuptake capacity in response to N supply in maize [21].In the present study,zmbri1roots showed lower expression levels ofZmNRT2.1andZmNRT2.2than wild-type roots under both LN and NN conditions.But eBL treatment increased the transcript expression ofZmNRT2.1andZmNRT2.2relative to the control.Thus,zmbri1roots showed lower transcription levels of NRT genes, leading to lowerfluxes and15uptake.These findings indicate that BRs could modulate the transcript expression of NRT genes to alter N uptake in maize in response to N supply.The regulatory interactions between BRs and NRT genes invite further study.

    5.Conclusions

    Fig.7.Changes in transcript expression of ZmNPF6.4, ZmNPF6.6, ZmNRT2.1, and ZmNRT2.2 in zmbri1 (a-d) and eBL-treated (e-h) roots under LN and NN conditions.The control and WT are maize cv.B73.Values with error bars are mean±SD(n=3).Different letters indicate significant difference between treatments at the same time point by Duncan’s multiple range teste at P < 0.05.eBL, 2,4-epibrassinolide; WT, wild type; LN, low N; NN, normal N.

    N supply influenced the transcript expression of genes involved in BR biosynthesis and signal transduction, and exogenous eBL application increased root length,plant biomass,and N accumulation in maize seedlings.zmbri1plants showed shorter roots and less plant biomass and N accumulation than wild-type plants under both LN and NN conditions.zmbri1plants showed lower netfluxes and15uptake than wild-type plants.The transcript expression ofZmNRTgenes was down-regulated inzmbri1roots relative to that in wild-type roots after N treatments.Exogenous application of eBL up-regulated the expression ofZmNRTgenes.Thus,BRs modulate N physiological response and regulate the transcript expression of NRT genes to promote N uptake in maize,suggesting a way to improve N efficiency in maize production.

    CRediT authorship contribution statement

    Jiapeng Xing:designed the research, performed experiments,and drafted the manuscript.Yubin Wang, Qingqing Yao, and Yushi Zhang:performed part of the work.Zhaohu Li:provided technical assistance.Mingcai Zhang:conceived the project and revised the manuscript.

    Declaration of competing interest

    The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.

    Acknowledgments

    This research was supported by National Key Research and Development Program of China (2017YFD0300410).We thank Dr.Philip W.Becraft and Dr.Yanhai Yin (Genetics, Development, and Cell Biology Department of Iowa State University,USA)for supplying theZmBRI1RNA interference (RNAi) mutants and excellent technical assistance.

    Appendix A.Supplementary data

    Supplementary data for this article can be found online at https://doi.org/10.1016/j.cj.2021.04.004.

    女人久久www免费人成看片| √禁漫天堂资源中文www| 国产精品一区二区三区四区免费观看| 亚洲少妇的诱惑av| 国产免费一区二区三区四区乱码| 99热国产这里只有精品6| 99视频精品全部免费 在线| 男人添女人高潮全过程视频| 黄色视频在线播放观看不卡| 在线观看国产h片| 日产精品乱码卡一卡2卡三| 高清视频免费观看一区二区| 18禁观看日本| 亚洲国产最新在线播放| 精品久久蜜臀av无| 久久久亚洲精品成人影院| 亚洲国产精品999| 十八禁高潮呻吟视频| 亚洲高清免费不卡视频| 国产精品人妻久久久影院| 午夜日本视频在线| 人妻制服诱惑在线中文字幕| 老熟女久久久| 精品人妻熟女av久视频| av天堂久久9| 这个男人来自地球电影免费观看 | 黄色毛片三级朝国网站| 五月伊人婷婷丁香| 美女脱内裤让男人舔精品视频| 久久国产精品男人的天堂亚洲 | 天美传媒精品一区二区| 亚洲国产精品专区欧美| 午夜福利在线观看免费完整高清在| 国产淫语在线视频| 在线亚洲精品国产二区图片欧美 | 少妇的逼好多水| 国产片内射在线| 五月玫瑰六月丁香| 午夜久久久在线观看| 久久人人爽av亚洲精品天堂| 国产日韩一区二区三区精品不卡 | 中文乱码字字幕精品一区二区三区| 99久久综合免费| 天堂8中文在线网| 多毛熟女@视频| 欧美国产精品一级二级三级| 夫妻午夜视频| 一级二级三级毛片免费看| √禁漫天堂资源中文www| 精品久久蜜臀av无| 七月丁香在线播放| 久久精品国产自在天天线| 亚洲天堂av无毛| 亚洲熟女精品中文字幕| 汤姆久久久久久久影院中文字幕| 欧美日韩一区二区视频在线观看视频在线| 精品少妇黑人巨大在线播放| 国产精品欧美亚洲77777| 99热网站在线观看| 日本色播在线视频| 国产熟女午夜一区二区三区 | 99久久综合免费| 久久99热这里只频精品6学生| 国产成人精品婷婷| 久久久久精品久久久久真实原创| freevideosex欧美| 免费高清在线观看日韩| 午夜福利,免费看| 欧美老熟妇乱子伦牲交| 久热这里只有精品99| av在线老鸭窝| 日日啪夜夜爽| 亚洲精品乱码久久久v下载方式| av在线播放精品| 亚洲av福利一区| 91aial.com中文字幕在线观看| 亚洲av欧美aⅴ国产| 久久免费观看电影| 国产免费一级a男人的天堂| 亚洲欧美一区二区三区黑人 | 欧美另类一区| √禁漫天堂资源中文www| 97超视频在线观看视频| 爱豆传媒免费全集在线观看| 久久久久精品久久久久真实原创| 中文欧美无线码| 三级国产精品欧美在线观看| 午夜福利在线观看免费完整高清在| 国产免费福利视频在线观看| 国产熟女欧美一区二区| 黄色配什么色好看| 大香蕉久久网| 久久精品国产亚洲av涩爱| 国产探花极品一区二区| 日日摸夜夜添夜夜添av毛片| 男女无遮挡免费网站观看| 亚洲av在线观看美女高潮| 高清在线视频一区二区三区| 精品人妻一区二区三区麻豆| 国产色爽女视频免费观看| 国产高清三级在线| 久久精品国产亚洲av天美| 美女视频免费永久观看网站| 精品人妻一区二区三区麻豆| 女的被弄到高潮叫床怎么办| 亚洲五月色婷婷综合| 精品一区二区三区视频在线| 欧美精品亚洲一区二区| av不卡在线播放| 久久 成人 亚洲| 啦啦啦视频在线资源免费观看| 天美传媒精品一区二区| 亚洲av电影在线观看一区二区三区| 搡老乐熟女国产| 久久久久人妻精品一区果冻| 久久精品国产自在天天线| av免费观看日本| 99精国产麻豆久久婷婷| 热99久久久久精品小说推荐| a 毛片基地| 午夜精品国产一区二区电影| 三级国产精品片| 国产精品99久久99久久久不卡 | 成人漫画全彩无遮挡| 黑人欧美特级aaaaaa片| 久久久久人妻精品一区果冻| 国产综合精华液| 色视频在线一区二区三区| 亚洲av电影在线观看一区二区三区| 日韩中文字幕视频在线看片| 国产国拍精品亚洲av在线观看| 又粗又硬又长又爽又黄的视频| 国产男女内射视频| 亚洲精品国产色婷婷电影| xxxhd国产人妻xxx| 美女内射精品一级片tv| 亚洲av免费高清在线观看| 在线免费观看不下载黄p国产| 国产精品久久久久久精品电影小说| 一级黄片播放器| 亚洲精品中文字幕在线视频| av线在线观看网站| 亚洲三级黄色毛片| 国产精品不卡视频一区二区| 亚洲久久久国产精品| 嫩草影院入口| 国内精品宾馆在线| 麻豆精品久久久久久蜜桃| 久久久久久久久久久丰满| av在线app专区| 丁香六月天网| 久热这里只有精品99| 高清视频免费观看一区二区| 久久精品人人爽人人爽视色| 三级国产精品欧美在线观看| freevideosex欧美| 制服诱惑二区| 免费不卡的大黄色大毛片视频在线观看| 一级a做视频免费观看| 在线观看国产h片| 高清不卡的av网站| 国产亚洲精品久久久com| 国产乱来视频区| 欧美成人午夜免费资源| 少妇的逼水好多| 日韩精品免费视频一区二区三区 | 黄色怎么调成土黄色| 91精品国产国语对白视频| 51国产日韩欧美| 中文字幕制服av| 老女人水多毛片| 少妇的逼水好多| 国产精品久久久久久久电影| 满18在线观看网站| 国产午夜精品久久久久久一区二区三区| 亚洲欧美成人综合另类久久久| 黑人高潮一二区| 一级二级三级毛片免费看| 青青草视频在线视频观看| 国产精品久久久久久av不卡| 国产精品熟女久久久久浪| 最近2019中文字幕mv第一页| 亚洲国产成人一精品久久久| 国产精品麻豆人妻色哟哟久久| 一级毛片aaaaaa免费看小| 亚洲国产精品999| 亚洲人成77777在线视频| 久久女婷五月综合色啪小说| 22中文网久久字幕| 日韩伦理黄色片| 国产精品人妻久久久影院| 久久ye,这里只有精品| 一区二区三区免费毛片| 另类精品久久| 97在线人人人人妻| 国产毛片在线视频| 日本黄大片高清| 国产一区二区三区av在线| 日韩制服骚丝袜av| 亚洲精品乱久久久久久| 国产高清国产精品国产三级| 高清不卡的av网站| av不卡在线播放| 免费看av在线观看网站| 国产亚洲av片在线观看秒播厂| 久热这里只有精品99| 高清视频免费观看一区二区| 成人亚洲欧美一区二区av| 亚洲av中文av极速乱| 久久女婷五月综合色啪小说| 国产女主播在线喷水免费视频网站| 亚洲精品,欧美精品| 最近最新中文字幕免费大全7| 中文字幕最新亚洲高清| 99九九在线精品视频| 九色亚洲精品在线播放| 亚洲av不卡在线观看| av在线app专区| 麻豆成人av视频| 欧美xxⅹ黑人| 女人精品久久久久毛片| 啦啦啦视频在线资源免费观看| av女优亚洲男人天堂| 国产日韩欧美亚洲二区| h视频一区二区三区| 亚洲人成77777在线视频| 亚洲精品日韩在线中文字幕| 亚洲av成人精品一二三区| 制服人妻中文乱码| 高清欧美精品videossex| 三级国产精品片| 丝袜喷水一区| 十八禁网站网址无遮挡| 狠狠婷婷综合久久久久久88av| 国产精品国产三级专区第一集| 久久久久精品久久久久真实原创| 免费av中文字幕在线| 一区二区三区乱码不卡18| 少妇熟女欧美另类| 日本91视频免费播放| 亚洲,欧美,日韩| 亚洲av欧美aⅴ国产| 日本vs欧美在线观看视频| 校园人妻丝袜中文字幕| 熟女人妻精品中文字幕| 日日啪夜夜爽| 91精品一卡2卡3卡4卡| 欧美三级亚洲精品| 又大又黄又爽视频免费| 搡老乐熟女国产| 日本欧美国产在线视频| 麻豆精品久久久久久蜜桃| 精品亚洲成国产av| 人妻一区二区av| 内地一区二区视频在线| 国产日韩欧美在线精品| 精品少妇黑人巨大在线播放| 永久网站在线| 视频在线观看一区二区三区| 寂寞人妻少妇视频99o| av播播在线观看一区| 一边亲一边摸免费视频| 一二三四中文在线观看免费高清| 丝袜脚勾引网站| 亚洲不卡免费看| 在线播放无遮挡| 午夜福利在线观看免费完整高清在| 26uuu在线亚洲综合色| 日韩强制内射视频| 夜夜骑夜夜射夜夜干| 91精品国产九色| 日韩欧美一区视频在线观看| 夫妻性生交免费视频一级片| 日本色播在线视频| 国产男女内射视频| 人妻人人澡人人爽人人| 亚洲国产欧美日韩在线播放| 午夜影院在线不卡| 五月开心婷婷网| 91国产中文字幕| 欧美国产精品一级二级三级| 国产精品久久久久成人av| 如何舔出高潮| 考比视频在线观看| 国产熟女欧美一区二区| 精品久久久久久久久亚洲| 午夜久久久在线观看| 亚洲国产最新在线播放| 另类精品久久| 日韩,欧美,国产一区二区三区| 国产欧美日韩综合在线一区二区| 亚洲,一卡二卡三卡| 一边摸一边做爽爽视频免费| 两个人免费观看高清视频| 天天躁夜夜躁狠狠久久av| 久久久精品94久久精品| 久久国内精品自在自线图片| 精品99又大又爽又粗少妇毛片| 看十八女毛片水多多多| 美女国产视频在线观看| √禁漫天堂资源中文www| 曰老女人黄片| 80岁老熟妇乱子伦牲交| 国产永久视频网站| 国产精品三级大全| 五月天丁香电影| 一级爰片在线观看| 精品熟女少妇av免费看| 一本—道久久a久久精品蜜桃钙片| 综合色丁香网| 欧美3d第一页| 国产成人一区二区在线| 亚洲美女黄色视频免费看| 国产高清有码在线观看视频| 18禁观看日本| 人体艺术视频欧美日本| 哪个播放器可以免费观看大片| 一级毛片电影观看| 老司机影院成人| 国产精品麻豆人妻色哟哟久久| 人人妻人人澡人人爽人人夜夜| 亚洲美女黄色视频免费看| av一本久久久久| 色婷婷久久久亚洲欧美| 十八禁高潮呻吟视频| 黄色一级大片看看| 男人操女人黄网站| 国产精品一区二区在线不卡| 高清欧美精品videossex| 亚洲国产成人一精品久久久| 亚洲人成网站在线观看播放| 肉色欧美久久久久久久蜜桃| 男人操女人黄网站| 久久青草综合色| 嘟嘟电影网在线观看| 国产精品不卡视频一区二区| 黄片无遮挡物在线观看| 天堂8中文在线网| 女的被弄到高潮叫床怎么办| 黄色一级大片看看| 亚洲欧美成人综合另类久久久| 卡戴珊不雅视频在线播放| 亚洲国产成人一精品久久久| 看免费成人av毛片| 久久99蜜桃精品久久| videossex国产| 纯流量卡能插随身wifi吗| 亚洲成人av在线免费| 日韩三级伦理在线观看| 美女国产高潮福利片在线看| 国产日韩欧美在线精品| 日韩一本色道免费dvd| 搡老乐熟女国产| 晚上一个人看的免费电影| 亚洲中文av在线| 国产成人91sexporn| 国内精品宾馆在线| 少妇被粗大的猛进出69影院 | 日韩中文字幕视频在线看片| 美女内射精品一级片tv| 精品99又大又爽又粗少妇毛片| 欧美+日韩+精品| 国产欧美日韩综合在线一区二区| 亚洲精品av麻豆狂野| 国产亚洲精品第一综合不卡 | 国产成人午夜福利电影在线观看| 亚洲精品,欧美精品| 午夜福利视频精品| 免费黄频网站在线观看国产| 大话2 男鬼变身卡| 97在线视频观看| 蜜桃国产av成人99| 在线观看免费视频网站a站| 国产黄色免费在线视频| 精品少妇久久久久久888优播| 国产精品久久久久久久久免| 国产伦理片在线播放av一区| 国产不卡av网站在线观看| 熟女av电影| 国产极品天堂在线| 97在线视频观看| 亚洲成人一二三区av| 国产欧美亚洲国产| 亚洲国产av影院在线观看| 国产精品熟女久久久久浪| 91精品一卡2卡3卡4卡| 国产av精品麻豆| 永久网站在线| 少妇的逼好多水| 亚洲精品国产av成人精品| 一级毛片aaaaaa免费看小| 超色免费av| 91精品伊人久久大香线蕉| 丝袜喷水一区| 亚洲精品自拍成人| av国产精品久久久久影院| 精品人妻一区二区三区麻豆| 我的老师免费观看完整版| 精品久久久久久久久av| 天堂俺去俺来也www色官网| 国产成人精品在线电影| 亚洲精品色激情综合| 国产乱来视频区| 久久久久久久久久人人人人人人| 国产极品粉嫩免费观看在线 | 亚洲精品av麻豆狂野| 纯流量卡能插随身wifi吗| 黄色一级大片看看| 国产成人免费观看mmmm| av女优亚洲男人天堂| 熟女av电影| 一区二区av电影网| 搡老乐熟女国产| 亚洲av综合色区一区| 99re6热这里在线精品视频| 久久精品国产自在天天线| 国语对白做爰xxxⅹ性视频网站| 国产成人精品一,二区| 亚洲精品自拍成人| 91在线精品国自产拍蜜月| 免费黄网站久久成人精品| 美女脱内裤让男人舔精品视频| 桃花免费在线播放| 一本一本综合久久| 午夜免费男女啪啪视频观看| 日本午夜av视频| 国产成人免费无遮挡视频| 国产色爽女视频免费观看| 永久免费av网站大全| 搡老乐熟女国产| 九九爱精品视频在线观看| 国产成人精品在线电影| 五月玫瑰六月丁香| 亚洲成人一二三区av| 久久女婷五月综合色啪小说| 国产精品人妻久久久影院| 狂野欧美白嫩少妇大欣赏| 亚洲在久久综合| 色吧在线观看| 在现免费观看毛片| 人妻一区二区av| 高清av免费在线| 飞空精品影院首页| 日本黄大片高清| 亚洲精品久久午夜乱码| 国产高清三级在线| 国产精品国产三级专区第一集| 亚洲成人一二三区av| 狠狠婷婷综合久久久久久88av| 99热这里只有是精品在线观看| 久久国产亚洲av麻豆专区| 欧美精品亚洲一区二区| 午夜免费观看性视频| 又大又黄又爽视频免费| 男人爽女人下面视频在线观看| 考比视频在线观看| 人人妻人人澡人人看| 97精品久久久久久久久久精品| 久久精品人人爽人人爽视色| 亚洲内射少妇av| 亚洲不卡免费看| 国产精品 国内视频| 校园人妻丝袜中文字幕| 91成人精品电影| 日韩人妻高清精品专区| 欧美日韩国产mv在线观看视频| 久久久欧美国产精品| 最新中文字幕久久久久| 国产精品一区www在线观看| 久久精品人人爽人人爽视色| 多毛熟女@视频| 亚洲av二区三区四区| 在线观看三级黄色| 香蕉精品网在线| 欧美精品一区二区大全| 国产精品久久久久久av不卡| 久久毛片免费看一区二区三区| 国产成人91sexporn| 韩国高清视频一区二区三区| 精品熟女少妇av免费看| 久久99热这里只频精品6学生| 中文字幕久久专区| 亚洲伊人久久精品综合| 亚洲国产欧美日韩在线播放| 在线天堂最新版资源| 国产精品三级大全| 午夜福利视频在线观看免费| a级毛片在线看网站| 一区二区三区乱码不卡18| 亚洲精华国产精华液的使用体验| 精品一区二区免费观看| 亚洲精品第二区| 国产av码专区亚洲av| 中文字幕最新亚洲高清| 自线自在国产av| 免费高清在线观看视频在线观看| 能在线免费看毛片的网站| 中国美白少妇内射xxxbb| 日韩中文字幕视频在线看片| 丝袜在线中文字幕| 亚洲精品乱码久久久久久按摩| 大片免费播放器 马上看| 国产精品不卡视频一区二区| 精品久久蜜臀av无| 狠狠精品人妻久久久久久综合| 亚洲精品视频女| 一本久久精品| 十八禁网站网址无遮挡| 久久久国产一区二区| 成人二区视频| 成人毛片a级毛片在线播放| 精品少妇内射三级| 中国美白少妇内射xxxbb| 久久人人爽人人爽人人片va| 欧美成人午夜免费资源| 大香蕉久久成人网| 老司机亚洲免费影院| 成人免费观看视频高清| av天堂久久9| 精品久久久噜噜| 国产伦精品一区二区三区视频9| 欧美丝袜亚洲另类| 亚洲精品乱码久久久久久按摩| 欧美 亚洲 国产 日韩一| 夫妻性生交免费视频一级片| 国产精品女同一区二区软件| 大又大粗又爽又黄少妇毛片口| 少妇人妻 视频| 妹子高潮喷水视频| 日韩一本色道免费dvd| 我要看黄色一级片免费的| 色94色欧美一区二区| 啦啦啦视频在线资源免费观看| 国产伦理片在线播放av一区| 国产男女内射视频| 久久久久久久亚洲中文字幕| 日韩不卡一区二区三区视频在线| 蜜桃国产av成人99| 亚洲精品,欧美精品| 欧美精品亚洲一区二区| 高清午夜精品一区二区三区| 高清不卡的av网站| 亚洲无线观看免费| 一级a做视频免费观看| 精品人妻一区二区三区麻豆| 五月伊人婷婷丁香| 99热这里只有是精品在线观看| 国产成人精品无人区| 性色avwww在线观看| 岛国毛片在线播放| 美女xxoo啪啪120秒动态图| 男女无遮挡免费网站观看| 亚洲av.av天堂| 欧美日韩亚洲高清精品| 亚洲精品亚洲一区二区| 在现免费观看毛片| 中文字幕人妻丝袜制服| av国产久精品久网站免费入址| 国产成人精品福利久久| 天堂8中文在线网| 麻豆成人av视频| 制服人妻中文乱码| 人妻 亚洲 视频| 欧美精品国产亚洲| 午夜福利视频在线观看免费| 少妇人妻久久综合中文| 纯流量卡能插随身wifi吗| 亚洲美女黄色视频免费看| 三级国产精品片| 亚洲精品乱码久久久久久按摩| 女性被躁到高潮视频| 中文字幕人妻丝袜制服| 能在线免费看毛片的网站| 日韩熟女老妇一区二区性免费视频| 观看美女的网站| 麻豆成人av视频| 国产日韩一区二区三区精品不卡 | 日韩一区二区三区影片| 黑丝袜美女国产一区| 黑人欧美特级aaaaaa片| 欧美激情 高清一区二区三区| 一级毛片电影观看| 一区二区日韩欧美中文字幕 | 岛国毛片在线播放| 国产高清有码在线观看视频| 精品人妻熟女毛片av久久网站| 色视频在线一区二区三区| 午夜福利,免费看| 9色porny在线观看| 国产不卡av网站在线观看| 搡老乐熟女国产| 亚洲欧美成人精品一区二区| 91久久精品国产一区二区三区| 久久久精品免费免费高清| 日韩av免费高清视频| 男女无遮挡免费网站观看| 日韩av免费高清视频| 成年女人在线观看亚洲视频| 国产片内射在线| 麻豆成人av视频| 国产av国产精品国产| 波野结衣二区三区在线| 国模一区二区三区四区视频| 全区人妻精品视频| 国产在视频线精品| 亚洲欧美一区二区三区黑人 | 免费看光身美女| 黄色毛片三级朝国网站| 国产精品.久久久| videossex国产| 精品国产露脸久久av麻豆| 欧美 亚洲 国产 日韩一| 久久99一区二区三区| 亚洲国产精品成人久久小说| 91久久精品电影网|