Identification of quantitative trait loci and epistasis for root characteristics and root exudations in maize (Zea mays L.) under deficient phosphorus

The phosphorus uptake （PU） in above-ground parts of plant, root characteristics and root exudations as well as the quantitative trait loci （QTLs） associated with these characteristics were determined for a F2：3 population derived from the cross of two contrasting maize （Zea mays L.） genotypes, 082 and Yel07. A total of 241 F2：3 families were evaluated in replicated trials under deficient phosphorus conditions in 2007 at two sites （Kaixian County and Southwest University, Chongqing, P. R. China）. The results show pleiotropy and close linkage among QTLs. Four common regions in different environments were in bnlg100- bnlg1268b （bins 1.02） for QTL of H＋, bnlg1268a-umc1290a （bins 1.09） for QTL of AP （acid phospbatase activity）, dupssrl5- P 1MT/a （bins 6.06） for QTLs of PU （phosphorus uptake） and RW （root weight）, and P IM3/d-P1M3/g （bins9.04） for QTLs of PU and AP. These QTLs are non-environment or minor QTLs x environment. By epistatic analysis, three main QTLs and eighteen QTLs x QTLs interactions were detected for the seven measured characteristics. These QTLs may affect trait expression by epistatic interaction with the other loci, and make a substantial contribution to phosphorus utilization efficiency, which should be considered when breeding maize varieties with high P efficiency. Two regions were detected in dupssrl 5- P1MT/a （bins 6.06） for QTL of RW and P1M3/d- P 1M3/g （bins 9.04） for QTL of PU and AP. They were detected in two different environments and by two methods of QTL analysis, which were useful for marker-assisted selection.

Analysis of TORCH results of retinal exudative changes in neonates

AIM:To explore the relationship between retinal exudative changes in neonates and perinatal toxoplasmosis,others,rubella,cytomegalovirus,and herpes simplex virus(TORCH)infections,as well as the characteristics of TORCH infection in neonates with retinal exudative changes.METHODS:Retrospective study.A total of 612 neonates with retinal exudative changes detected during ophthalmic screening in our hospital from May 2019 to March 2023 were selected.TORCH tests were performed on these neonates,and the results were subjected to statistical analysis to determine the infection characteristics.The neonates with retinal exudative changes were grouped by sex and age,the characteristics of TORCH infection were analyzed,and the positive rates were compared.RESULTS:Among the 612 neonates with retinal exudative changes,the highest positive rate was observed for cytomegalovirus(CMV-IgG)(96.7%),followed by rubella virus(RV-IgG)(73.9%).Mixed infections with two or three viruses were also observed,with the highest positive rate for mixed infection of RV-IgG and CMV-IgG reaching 71.2%.There was no statistically significant difference in TORCH infection among neonates of different sex(P>0.05).However,there were statistically significant differences in RV-IgG and CMV-IgM infections with retinal exudative changes among neonates of different age groups(P<0.05).CONCLUSION:Perinatal TORCH infection may be an important factor causing retinal exudative changes in neonates.The differences in various infections are not related to sex but are related to different age groups.

De novel heterozygous copy number deletion on 7q31.31-7q31.32 involving TSPAN12 gene with familial exudative vitreoretinopathy in a Chinese family

AIM:To investigate the genetic and clinical characteristics of patients with a large heterozygous copy number deletion on 7q31.31-7q31.32.METHODS:A family with familial exudative vitreoretinopathy(FEVR)phenotype was included in the study.Whole-exome sequencing(WES)was initially used to locate copy number variations(CNVs)on 7q31.31-31.32,but failed to detect the precise breakpoint.The long-read sequencing,Oxford Nanopore sequencing Technology(ONT)was used to get the accurate breakpoint which is verified by quantitative real-time polymerase chain reaction(QPCR)and Sanger Sequencing.RESULTS:The proband,along with her father and younger brother,were found to have a heterozygous 4.5 Mb CNV deletion located on 7q31.31-31.32,which included the FEVRrelated gene TSPAN12.The specific deletion was confirmed as del(7)(q31.31q31.32)chr7:g.119451239_123956818del.The proband exhibited a phase 2A FEVR phenotype,characterized by a falciform retinal fold,macular dragging,and peripheral neovascularization with leaking of fluorescence.These symptoms led to a significant decrease in visual acuity in both eyes.On the other hand,the affected father and younger brother showed a milder phenotype.CONCLUSION:The heterozygous CNV deletion located on 7q31.31-7q31.32 is associated with the FEVR phenotype.The use of long-read sequencing techniques is essential for accurate molecular diagnosis of genetic disorders.

IM-EDRD from Retinal Fundus Images Using Multi-Level Classification Techniques

In recent years,there has been a significant increase in the number of people suffering from eye illnesses,which should be treated as soon as possible in order to avoid blindness.Retinal Fundus images are employed for this purpose,as well as for analysing eye abnormalities and diagnosing eye illnesses.Exudates can be recognised as bright lesions in fundus pictures,which can be thefirst indicator of diabetic retinopathy.With that in mind,the purpose of this work is to create an Integrated Model for Exudate and Diabetic Retinopathy Diagnosis(IM-EDRD)with multi-level classifications.The model uses Support Vector Machine(SVM)-based classification to separate normal and abnormal fundus images at thefirst level.The input pictures for SVM are pre-processed with Green Channel Extraction and the retrieved features are based on Gray Level Co-occurrence Matrix(GLCM).Furthermore,the presence of Exudate and Diabetic Retinopathy(DR)in fundus images is detected using the Adaptive Neuro Fuzzy Inference System(ANFIS)classifier at the second level of classification.Exudate detection,blood vessel extraction,and Optic Disc(OD)detection are all processed to achieve suitable results.Furthermore,the second level processing comprises Morphological Component Analysis(MCA)based image enhancement and object segmentation processes,as well as feature extraction for training the ANFIS classifier,to reliably diagnose DR.Furthermore,thefindings reveal that the proposed model surpasses existing models in terms of accuracy,time efficiency,and precision rate with the lowest possible error rate.

A Novel Soft Clustering Method for Detection of Exudates

One of the earliest indications of diabetes consequence is Diabetic Retinopathy(DR),the main contributor to blindness worldwide.Recent studies have proposed that Exudates(EXs)are the hallmark of DR severity.The present study aims to accurately and automatically detect EXs that are difficult to detect in retinal images in the early stages.An improved Fusion of Histogram-Based Fuzzy C-Means Clustering(FHBFCM)by a New Weight Assignment Scheme(NWAS)and a set of four selected features from stages of pre-processing to evolve the detection method is proposed.The features of DR train the optimal parameter of FHBFCM for detecting EXs diseases through a stepwise enhancement method through the coarse segmentation stage.The histogram-based is applied to find the color intensity in each pixel and performed to accomplish Red,Green,and Blue(RGB)color information.This RGB color information is used as the initial cluster centers for creating the appropriate region and generating the homogeneous regions by Fuzzy C-Means(FCM).Afterward,the best expression of NWAS is used for the delicate detection stage.According to the experiment results,the proposed method successfully detects EXs on the retinal image datasets of DiaretDB0(Standard Diabetic Retinopathy Database Calibration level 0),DiaretDB1(Standard Diabetic Retinopathy Database Calibration level 1),and STARE(Structured Analysis of the Retina)with accuracy values of 96.12%,97.20%,and 93.22%,respectively.As a result,this study proposes a new approach for the early detection of EXs with competitive accuracy and the ability to outperform existing methods by improving the detection quality and perhaps significantly reducing the segmentation of false positives.

Efficacy of lidocaine wet compress combined with red-light irradiation for chronic wounds

BACKGROUND Chronic wounds that fail to progress through normal phases of healing present a significant healthcare burden owing to prolonged treatment and associated costs.Traditional wound care typically involves regular dressing changes,which can be painful.Recent approaches have explored the use of lidocaine to manage pain and red-light irradiation(RLI),known for its anti-inflammatory and cell proliferation effects,to potentially enhance wound healing.AIM To investigate the therapeutic efficacy of lidocaine wet compression(LWC)combined with RLI for chronic wounds.METHODS We enrolled 150 patients with chronic wounds from the Wound and Ostomy Outpatient Clinic of the Second Hospital of Anhui Medical University from April to September 2022.The wounds were treated with dressing changes.The patients were randomly assigned to the control and experimental groups using a random number table and given the same first dressing change(2%LWC for 5 min and routine dressing change).From the second dressing change,in addition to 2%LWC for 5 min and routine dressing change,the experimental group received RLI,whereas the control group continued to receive the same LWC and dressing change.The first and second dressing changes were performed on days 1 and 2,respectively.The third dressing change was performed 3 d after the second change.The frequency of subsequent dressing changes was determined based on wound exudation and pain.Pain during the first three dressing changes was evaluated in both groups.The average number of dressing changes within 28 d and the degree of wound healing on day 28 were also recorded.RESULTS During the initial dressing change,no noticeable differences were observed in the pain levels experienced by the two groups,indicating similar pain tolerance.However,during the second and third dressing changes,the experimental group reported significantly less pain than the control group.Furthermore,over 28 d,the experimental group required fewer dressing changes than the control group.CONCLUSION Notably,the effectiveness of wound healing on the 28th day was significantly higher in the experimental group than that of in the control group.The combination of LWC and RLI was effective in reducing early-stage pain,promoting wound healing,decreasing the frequency of dressing changes,and enhancing patients’overall quality of life with chronic wounds.

蟛蜞菊根分泌物的异种克生作用及初步分离

蟛蜞菊根分泌物的异种克生作用及初步分离曾任森，林象联，谭惠芬，曾强（华南农业大学农业生态室，广州５１０６４２）（南开大学元素有机化学研究所，天津３０００７）ＡｌｌｅｌｏｐａｔｈｉｃＥｆｆｅｃｔｓａｎｄＰｒｅｌｉｍｉｎａｒｙＩｓｏｌａｔｉｏｎｏｆＲｏｏ...

Preliminary Study on Garlic Root Exudates Influences to the Growth of Pomegranate Wilt Pathogen(Ceratocystis fimbriata)and Bacillus subtilis

[Objectives] This paper aims to explore the possibility to intercrop garlic with pomegranate tree to control pomegranate wilt.[Methods] Root exudates of garlic is cultivated in 1/5 concentration of MS solution and distilled water is examined in lab to test their effect to growth of mycelia of pomegranate wilt pathogen（Ceratocystis fimbriata）and multiplication of Bacillus subtilis.[Results] The result shows that garlic root exudates whatever cultivated in MS solution or distilled water could not inhibit or promote mycelia growth of C.fimbriata.However,garlic root exudates cultivated in both methods effectively promote multiplication of B.subtilis.[Conclusions] It is suggested that intercropping garlic with pomegranate tree by combining application B.subtilis could be a promising way to prevent pomegranate wilt spread in practice.

Analysis of the major components of root exudates released from several economic forest tree using GC-MS

In 2000 and 2001, 1-year-old seedlings of 7 economic tree species including chestnut, apricot, persimmon, peach, walnut, pear and apple were cultured in garden pots that had a diameter of 40 cm and were filled with clean sand. The major components of exudates released from their roots were isolated and analysed by GC-MS. Totally 200 kinds of organic chemicals were isolated, of which 3 kinds i.e. naphthalene, dimethylbenzene and dibutyl phthalate were principally controlled pollutants according to 'Blacklist of Principal Environment Pollutants in China' and the standard of U.S. Environmental Protection Agency (EPA). The research result provided theoretical evidence for selecting low-pollution economic forest crops in the water source protection area in Miyun Reservoir.

The Effects of Climate,Soil and Cultivar on the Content of Surface Trichome Exudates from Fresh Flue-cured Leaves

[Objective] The aim was to study the effect of climate and soil on the content of surface trichome exudates from fresh flue-cured leaves. [Method] The experiment was conducted in 2009 in Longgang, Weining and Tianzhu of Guizhou Province with flue-cured tobacco variety Yun 85. Mature tobacco leaves were taken from lower, middle and upper plant positions respectively and major cuticular com-ponents of the leaves were extracted with methylene chloride and their chemical compositions were analyzed with GC-MS. [Result] The result showed that there was large difference among trichome exudates of leaves grown under different climatic conditions. The leaf trichome exudates from Tianzhu were quite higher than those from Weining and Longgang, and trichome exudates difference between Tianzhu and Longgang was significant. Soil conditions had considerable effect on the content of tobacco leaf surface trichome exudates, leaf from Longgang soil had highest con-tent, fol owed by Tianzhu soil leaf, Weining soil leaf had lowest content. Under vari-ous climatic conditions, the contents of leaf trichome exudates of same soil were in-consistent, indicating the presence of certain climate and soil factor interaction; The contents of leaf trichome exudates from four cultivars in Longgang showed large difference, Nanjiang 3 was the lowest, fol owed by K326, Guiyan 201 was high, and Yunyan85 the highest. The content of leaf trichome exudates of Yunyan85 was more than two times of that of Nanjiang 3. [Conclusion] The Effect of climate factor on the content of leaf trichome exudates was greater than that of cultivar, which was then larger than soil condition.

Root Exudates, Rhizosphere Zn Fractions, and Zn Accumulation of Ryegrass at Different Soil Zn Levels

A glasshouse experiment was conducted using a root-bag technique to study the root exudates, rhizosphere Zn fractions, and Zn concentrations and accumulations of two ryegrass cultivars （Lolium perenne L. cvs. Airs and Tede） at different soil Zn levels （0, 2, 4, 8, and 16 mmol kg^-1 soil）. Results indicated that plant growth of the two cultivars was not advérsely affected at soil Zn level ≤ 8 mmol kg^-1. Plants accumulated more Zn as soil Zn levels increased, and Zn concentrations of shoots were about 540 μg g^-1 in Aris and 583.9 μg g^-1 in Tede in response to 16 mmol Zn kg^-1 soil. Zn ratios of shoots to roots across the soil Zn levels were higher in Tede than in Airs, corresponding with higher rhizosphere available Zn fractions （exchangeable, bound to manganese oxides, and bound to organic matter） in Airs than in Tede. Low-molecular-weight （LMW） organic acids （oxalic, tartaric, malic, and succinic acids） and amino acids （proline, threonine, glutamic acid, and aspartic acid, etc.） were detected in root exudates, and the concentrations of LMW organic acids and amino acids increased with addition of 4 mmol Zn kg^-1 soil compared with zero Zn addition. Higher rhizosphere concentrations of oxalic acid, glutamic acid, alanine, phenylalanine, leucine, and proline in Tede than in Airs likely resulted in increased Zn uptake from the soil by Tede than by Airs. The results suggested that genotypic differences in Zn accumulations were mainly because of different root exudates and rhizosphere Zn fractions.

Effects of maize root exudates and organic acids on the desorption of phenanthrene from soils

The effects of maize root exudates and low-molecular-weight-organic anions （LMWOAs） on the desorption of phenanthrene from eight artificially contaminated soils were evaluated. A significant negative correlation was observed between the amounts of phenanthrene desorbed and the soil organic carbon （SOC） contents （P 〈 0.01）, and the influences of soil pH and clay content on phenanthrene desorption were insignificant （P 〉 0.1）. Neither maize root exudates nor oxalate and citrate anions influenced desorption of phenanthrene with the addition of NaN3. A faster phenanthrene desorption occurred without the addition of NaN3 in the presence of maize root exudates than oxalate or citrate due to the enhanced degradation by root exudates. Without the addition of NAN3, oxalate or citrate at different concentrations could inhibit phenanthrene desorption to different extents and the inhibiting effect by citrate was more significant than by oxalate. This study leads to the conclusion that maize root exudates can not enhance the desorption under abiotic condition with the addition of NaN3 and can promote the desorption of phenanthrene in soils without the addition of NaN3.

Influence of Indian mustard (Brassica juncea) on rhizosphere soil solution chemistry in long-term contaminated soils:A rhizobox study

This study investigated the influence of Indian mustard （Brassicajuncea） root exudation on soil solution properties （pH, dissolved organic carbon （DOC）, metal solubility） in the rhizosphere using a rhizobox. Measurement was conducted following the cultivation of Indian mustard in the rhizobox filled four different types of heavy metal contaminated soils （two alkaline soils and two acidic soils）. The growth of Indian mustard resulted in a significant increase （by 0.6 pH units） in rhizosphere soil solution pH of acidic soils and only a slight increase （〈 0.1 pH units） in alkaline soils. Furthermore, the DOC concentration increased by 17-156 mg/L in the rhizosphere regardless of soil type and the extent of contamination, demonstrating the exudation of DOC from root. Ion chromatographic determination showed a marked increase in the total dissolved organic acids （OAs） in rhizosphere. While root exudates were observed in all soils, the amount of DOC and OAs in soil solution varied considerably amongst different soils, resulting in significant changes to soil solution metals in the rhizosphere. For example, the soil solution Cd, Cu, Pb, and Zn concentrations increased in the rhizosphere of alkaline soils compared to bulk soil following plant cultivation. In contrast, the soluble concentrations of Cd, Pb, and Zn in acidic soils decreased in rhizosphere soil when compared to bulk soils. Besides the influence of pH and DOC on metal solubility, the increase of heavy metal concentration having high stability constant such as Cu and Pb resulted in a release of Cd and Zn from solid phase to liquid phase.

Soluble protein and acid phosphatase exuded by ectomycorrhizal fungi and seedlings in response to excessive Cu and Cd

Fungi and their symbionts can alleviate heavy metal stress by exuding soluble proteins and enzymes. This study examined the role of soluble protein and acid phosphatase （APase） exuded by Xerocomus chrysenteron, an ectomycorrhizal fungus, and the seedlings of its symbiont, Chinese pine （Pinus tabulaeformis）, under conditions of excessive Cu and Cd. The growth type showed that this poorly studied ectomycorrhizal fungus was capable of tolerating high concentrations of Cu, and may be useful in phytoremediation. X. chrysenteron grew well at 80 mg/L Cu, and the EC50 for Cd was 17.82 mg/L. X. chrysenteron also showed enhanced exudation of soluble protein in both isolated and inoculated cultivations under the influence of Cu and Cd. Soluble protein exudation, however, differed under Cu and Cd stress in isolates. In mediums containing Cu, soluble protein exudation increased with concentration, but in mediums containing Cd the content of soluble protein increased to a comparable level at all concentrations. This study demonstrated that soluble protein was related to heavy metal tolerance, although the different ions played different roles. While APase activity in exudates of fungi and seedlings decreased under Cu and Cd stress in comparison to the control, the APase activity in seedlings was maintained by inoculation. Thus, X. chrysenteron facilitated the ability of plant to maintain a normal nutrient uptake, and therefore to protect it from heavy metal toxicity.

Artificial root exudates and soil organic carbon mineralization in a degraded sandy grassland in northern China

Plant root exudates contain various organic and inorganic components that include glucose, citric and oxalic acid. These components affect rhizosphere microbial and microfaunal activities, but the mechanisms are not fully known. Studies concerned from degraded grassland ecosystems with low soil carbon（C） contents are rare, in spite of the global distribution of grasslands in need of restoration. All these have a high potential for carbon sequestration, with a reduced carbon content due to overutilization. An exudate component that rapidly decomposes will increase soil respiration and CO2 emission, while a component that reduces decomposition of native soil carbon can reduce CO2 emission and actually help sequestering carbon in soil. Therefore, to investigate root exudate effects on rhizosphere activity, citric acid, glucose and oxalic acid（0.6 g C/kg dry soil） were added to soils from three biotopes（grassland, fixed dune and mobile dune） located in Naiman, Horqin Sandy Land, Inner Mongolia, China） and subjected to a 24-day incubation experiment together with a control. The soils were also analyzed for general soil properties. The results show that total respiration without exudate addition was highest in grassland soil, intermediate in fixed dune and lowest in mobile dune soil. However, the proportion of native soil carbon mineralized was highest in mobile dune soil, reflecting the low C/N ratio found there. The exudate effects on CO2-C emissions and other variables differed somewhat between biotopes, but total respiration（including that from the added substrates） was significantly increased in all combinations compared with the control, except for oxalic acid addition to mobile dune soil, which reduced CO2-C emissions from native soil carbon. A small but statistically significant increase in pH by the exudate additions in grassland and fixed dune soil was observed, but there was a major decrease from acid additions to mobile dune soil. In contrast, electrical conductivity decreased in grassland and fixed dune soil and increased in mobile dune. Thus, discrete components of root exudates affected soil environmental conditions differently, and responses to root exudates in soils with low carbon contents can differ from those in normal soils. The results indicate a potential for, e.g., acid root exudates to decrease decomposition rate of soil organic matter in low carbon soils, which is of interest for both soil restoration and carbon sequestration.

Effect of Organic Acids and Protons on Release of Non-Exchangeable NH_4^+ in Flooded Paddy Soils

In a model experiment, which imitated the rhizosphere of rice, the effect of organic acids (oxalic acid, citric acid) and protons on the release of non-exchangeable NH4+ and the resin adsorption of N was studied in a paddy soil, typical for Zhejiang Province, China. Oxalic and citric acids under low pH conditions, in combination with proton secretion, favored the mobilization of NH4+ ions and increased resin adsorption of N. The release of non-exchangeable NH4+ was associated with less formation of iron oxides. These could coat clay minerals and thus hinder the diffusion of NH4+ ions out of the interlayer. Protons enhanced the release of NH4+, and then they could enter the wedge zones of the clay minerals and displace non-exchangeable NH4+ ions.

Microbial activity related to N cycling in the rhizosphere of maize stressed by heavy metals

A greenhouse experiment was carried out to compare differences in potential activities of ammonification, nitrification and denitrification in rhizosphere and bulk soil in a heavy-metal-stressed system. Exchangeable fractions of Cd, Cu and Cr were all higher in the rhizosphere of maize than in bulk soil. Results showed that the mineralization of N in soil was stimulated by low concentration of Cd. Addition of Cd at low levels stimulated the ammonifying and nitrifying activity in soil, while inhibitory influences were shown at high levels. Nitrifying bacteria was proved to be the most sensitive one, whilst the effect on denitrifying bacteria was very limited. Comparing Cd, Cu and Cr(VI) at 20 mg/kg soil, Cd was the most effective inhibitor of ammonification and denitrification, while Cr(VI) had the strongest inhibitory influence on nitrifying activity. Root exudates played important roles on the different exchangeable metal fractions and bacterial activities between rhizosphere and non-rhizosphere. Nitrate was the main form of mineral N in soil, as well as the main form of N absorbed by plants, but the formation and relative absorption of ammonium were promoted in response to high Cd exposure.

Overexpression of a Foreign Bt Gene in Cotton Affects the Low-Molecular-Weight Components in Root Exudates

Most research in the past using genetically modified crops (GM crops) has focused on the ecological safety of foreign gene (i.e., the gene flow), gene products (for example, Bt (Bacillus thuringiensis) protein), and the safety of transgenic food for humans. In this study, changes in both the species and amounts of low-molecular-weight components in cotton (Gossypium hirsutum L.) root exudates after foreign Bt gene overexpression were investigated under different nutritional conditions. Transgenic cotton containing Bt (Bt-cotton), supplemented with all the mineral nutrients, secreted more organic acids than the wild-type cotton (WT). When nitrogen was removed from the full-nutrient solution, the amount of organic acids secretion of Bt-cotton was lesser than that of WT. The roots of the transgenic cotton secreted lesser amounts of amino acids and soluble sugars than the WT roots in the full-nutrient solution. Deficiencies of P and K caused a large increase in the total amino acid and soluble sugar secretions of both Bt-cotton and WT, with larger increases observed in Bt-cotton. Because transferring the foreign Bt gene into cotton can result in alterations in the components of the root exudates, with the effect varying depending on the nutritional status, the cultivation of genetically modified crops, such as Bt-cotton, in soil environments should be more carefully assessed, and the possible effects as a result of the alterations in the root exudate components should be considered.

The Extraction,Isolation and Identification of Exudates from the Roots of Flaveria bidentis

Large amounts ofFlaveria bidentis＇s root culturing solution were obtained by using DFT （deep flow technique） equipment and these solution which was vacuum concentrated （10, 20 mg mL-~） can have a certain inhibition on Triticum aestivum, Cucumis sativus, Raphanus sativus, Amaranthus retroflexus, Setaria viridis, Chenopodium album, Echinochloa crusgalli and Chloris virgata. This outcome suggested some active compounds in the root exudates ofFIaveria bidentis can inhibit the germination, seedling elongation and root length. The dichloromethane extract of root exudates was identificated by GC-MS, and 29 kinds of compounds, including esters, hydrocarbons, ketones, thiazole, amines, etc. were obtained and the phthalate n-octyl ester, phthalate 2-ethylhexyl ester were proved to be allelochemicals. The culturing solution of root exudates was separated through the resin column and silica gel column and a component inhibiting seedling height, root length and fresh weight of wheat was got. There were 6 kinds of organic compounds in this component including dioctyl phthalate, 1,2-phthalate, mono（2-ethylhexyl） ester by GC-MS.

Determination of Organic Acids in Root Exudates by High Performance Liquid Chromatography: I. Development and Assessment of Chromatographic Conditions *1

Methods for determining nine low molecular weight organic acids in root exudates were developed by using reversed phase high performance liquid chromatography with UV (ultraviolet) detection at 214 nm. The mobile phase was 18 mmol L -1 kH 2PO 4 adjusted to pH 2.25 with phosphoric acid and the flow rate was 0.3 mL min -1 . The analytical column was a reversed phase silica based C 18 column (Shim pack CLC ODS). The root exudates were collected through submerging the whole root system into aerated deionized water for 2 hours. The filtered exudate solutions were concentrated to dryness by rotary evaporation at 40 °C, dissolved in 10 mL mobile phase. The chromatographic conditions of organic acid determination were analyzed. The results showed that there was a high selectivity and sensitivity in the organic acid determination by reversed phase high performance liquid chromatography. Coefficients of variation for organic acid determination were lower than 10% except lactic acid. The recoveries were consistently between 80.1% to 108.3%. Detection limits were approximately 0.05 to 4.5 mg L -1 for organic acids except succinic acid with the detection limit of 7.0 mg L -1 . Phosphorus deficiency may contribute to the release of organic acids in soybean root exudates especially malic, lactic and citric acids.