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1.  Disorganized distribution of homogalacturonan epitopes in cell walls as one possible mechanism for aluminium-induced root growth inhibition in maize 
Annals of Botany  2009;104(2):235-241.
Background and Aims
Aluminium (Al) toxicity is one of the most severe limitations to crop production in acid soils. Inhibition of root elongation is the primary symptom of Al toxicity. However, the underlying basis of the process is unclear. Considering the multiple physiological and biochemical functions of pectin in plants, possible involvement of homogalacturonan (HG), one of the pectic polysaccharide domains, was examined in connection with root growth inhibition induced by Al.
Methods
An immunolabelling technique with antibodies specific to HG epitopes (JIM5, unesterified residues flanked by methylesterifed residues; JIM7, methyl-esterified residues flanked by unesterified residues) was used to visualize the distribution of different types of HG in cell walls of root apices of two maize cultivars differing in Al resistance.
Key Results
In the absence of Al, the JIM5 epitope was present around the cell wall with higher fluorescence intensity at cell corners lining the intercellular spaces, and the JIM7 epitope was present throughout the cell wall. However, treatment with 50 µm Al for 3 h produced 10 % root growth inhibition in both cultivars and caused the disappearance of fluorescence in the middle lamella of both epitopes. Prolonged Al treatment (24 h) with 50 % root growth inhibition in ‘B73’, an Al-sensitive cultivar, resulted in faint and irregular distribution of both epitopes. In ‘Nongda3138’, an Al-resistant cultivar, the distribution of HG epitopes was also restricted to the lining of intercellular spaces when a 50 % inhibition to root growth was induced by Al (100 µm Al, 9 h). Altered distribution of both epitopes was also observed when of roots were exposed to 50 µm LaCl3 for 24 h, resulting in 40 % inhibition of root growth.
Conclusions
Changes in HG distribution and root growth inhibition were highly correlated, indicating that Al-induced perturbed distribution of HG epitopes is possibly involved in Al-induced inhibition of root growth in maize.
doi:10.1093/aob/mcp123
PMCID: PMC2710910  PMID: 19483201
Al toxicity; cell wall; homogalacturnonan; immunofluorescence; methylesterification; pectin
2.  The role of the root apoplast in aluminium-induced inhibition of root elongation and in aluminium resistance of plants: a review 
Annals of Botany  2010;106(1):185-197.
Background
Aluminium (Al) toxicity is the most important soil constraint for plant growth and development in acid soils. The mechanism of Al-induced inhibition of root elongation is still not well understood, and it is a matter of debate whether the primary lesions of Al toxicity are apoplastic or symplastic.
Scope
The present review focuses on the role of the apoplast in Al toxicity and resistance, summarizing evidence from our own experimental work and other evidence published since 1995.
Conclusions
The binding of Al in the cell wall particularly to the pectic matrix and to the apoplastic face of the plasma membrane in the most Al-sensitive root zone of the root apex thus impairing apoplastic and symplastic cell functions is a major factor leading to Al-induced inhibition of root elongation. Although symplastic lesions of Al toxicity cannot be excluded, the protection of the root apoplast appears to be a prerequisite for Al resistance in both Al-tolerant and Al-accumulating plant species. In many plant species the release of organic acid anions complexing Al, thus protecting the root apoplast from Al binding, is a most important Al resistance mechanism. However, there is increasing physiological, biochemical and, most recently also, molecular evidence showing that the modification of the binding properties of the root apoplast contributes to Al resistance. A further in-depth characterization of the Al-induced apoplastic reaction in the most Al-sensitive zone of the root apex is urgently required, particularly to understand the Al resistance of the most Al-resistant plant species.
doi:10.1093/aob/mcq053
PMCID: PMC2889789  PMID: 20237112
Aluminium; aluminum; resistance; apoplast; cell wall; pectin; root elongation
3.  Aluminium localization in root tips of the aluminium-accumulating plant species buckwheat (Fagopyrum esculentum Moench) 
Journal of Experimental Botany  2011;62(15):5453-5462.
Aluminium (Al) uptake and transport in the root tip of buckwheat is not yet completely understood. For localization of Al in root tips, fluorescent dyes and laser ablation inductively coupled plasma mass spectrometry (LA-ICP-MS) were compared. The staining of Al with morin is an appropriate means to study qualitatively the radial distribution along the root tip axis of Al which is complexed by oxalate and citrate in buckwheat roots. The results compare well with the distribution of total Al determined by LA-ICP-MS which could be reliably calibrated to compare with Al contents by conventional total Al determination using graphite furnace atomic absorption spectrometry. The Al localization in root cross-sections along the root tip showed that in buckwheat Al is highly mobile in the radial direction. The root apex predominantly accumulated Al in the cortex. The subapical root section showed a homogenous Al distribution across the whole section. In the following root section Al was located particularly in the pericycle and the xylem parenchyma cells. With further increasing distance from the root apex Al could be detected only in individual xylem vessels. The results support the view that the 10 mm apical root tip is the main site of Al uptake into the symplast of the cortex, while the subapical 10–20 mm zone is the main site of xylem loading through the pericycle and xylem parenchyma cells. Progress in the better molecular understanding of Al transport in buckwheat will depend on the consideration of the tissue specificity of Al transport and complexation.
doi:10.1093/jxb/err222
PMCID: PMC3223043  PMID: 21831842
Aluminum; Al accumulator; Al–organic acid complex; in situ Al quantification; laser ablation ICP-MS; lumogallion; morin; xylem loading
4.  Differential transcript profiling through cDNA-AFLP showed complexity of rutin biosynthesis and accumulation in seeds of a nutraceutical food crop (Fagopyrum spp.) 
BMC Genomics  2012;13:231.
Background
Buckwheat, consisting of two cultivated species Fagopyrum tataricum and F. esculentum, is the richest source of flavonoid rutin. Vegetative tissues of both the Fagopyrum species contain almost similar amount of rutin; however, rutin content in seed of F. tataricum are ~50 folds of that in seed of F. esculentum. In order to understand the molecular basis of high rutin content in F. tataricum, differential transcript profiling through cDNA-AFLP has been utilized to decipher what genetic factors in addition to flavonoid structural genes contribute to high rutin content of F. tataricum compared to F. esculentum.
Results
Differential transcript profiling through cDNA-AFLP in seed maturing stages (inflorescence to seed maturation) with 32 primer combinations generated total of 509 transcript fragments (TDFs). 167 TDFs were then eluted, cloned and sequenced from F. tataricum and F. esculentum. Categorization of TDFs on the basis of their presence/absence (qualitative variation) or differences in the amount of expression (quantitative variation) between both the Fagopyrum species showed that majority of variants are quantitative (64%). The TDFs represented genes controlling different biological processes such as basic and secondary metabolism (33%), regulation (18%), signal transduction (14%), transportation (13%), cellular organization (10%), and photosynthesis & energy (4%). Most of the TDFs except belonging to cellular metabolism showed relatively higher transcript abundance in F. tataricum over F. esculentum. Quantitative RT-PCR analysis of nine TDFs representing genes involved in regulation, metabolism, signaling and transport of secondary metabolites showed that all the tested nine TDFs (Ubiquitin protein ligase, ABC transporter, sugar transporter) except MYB 118 showed significantly higher expression in early seed formation stage (S7) of F. tataricum compared to F. esculentum. qRT-PCR results were found to be consistent with the cDNA-AFLP results.
Conclusions
The present study concludes that in addition to structural genes, other classes of genes such as regulators, modifiers and transporters are also important in biosynthesis and accumulation of flavonoid content in plants. cDNA-AFLP technology was successfully utilized to capture genes that are contributing to differences in rutin content in seed maturing stages of Fagopyrum species. Increased transcript abundance of TDFs during transition from flowers to seed maturation suggests their involvement not only in the higher rutin content of F. tataricum over F. esculentum but also in nutritional superiority of the former.
doi:10.1186/1471-2164-13-231
PMCID: PMC3441755  PMID: 22686486
Rutin; cDNA-AFLP; Transcript derived fragments; qRT-PCR
5.  Genetic analyses of agronomic traits in Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) 
Breeding Science  2012;62(4):303-309.
The consumption of products made from Tartary buckwheat (Fagopyrum tataricum (L.) Gaertn.) has increased in recent years in Japan. Increased consumer demand has led to recognition of the need for early varieties of this crop with high and stable yields. In order to accomplish this, more information is needed on the genetic mechanisms affecting earliness and yield. We conducted genetic analysis of 3 agronomic traits (days to flowering, plant height and total seed weight per plant) to segregate F2 and F3 populations derived from a cross between Tartary buckwheat cultivars ‘Hokuriku No. 4’ and ‘Ishisoba’. Broad-sense heritability estimates for days to flowering, plant height and total seed weight were 0.70, 0.62 and 0.75, respectively, in F3 population. Narrow-sense heritability for total seed weight (0.51) was highest, followed by heritability for days to flowering (0.37), with heritability for plant height (0.26) lowest. Later flowering was associated with increased plant height and higher yields. From the F4 generation, we identified twelve candidate plants with earlier maturity and reduced plant height compared to ‘Hokuriku No. 4’, but almost the same total seed weight. These results suggest that hybridization breeding using the single seed descent (SSD) method is an effective approach for improving agronomic characteristics of Tartary buckwheat.
doi:10.1270/jsbbs.62.303
PMCID: PMC3528326  PMID: 23341743
Tartary buckwheat; segregating generation; genetic parameter; heritability; genetic correlation; phenotypic correlation; hybridization breeding
6.  Physiological and molecular analysis of the interaction between aluminium toxicity and drought stress in common bean (Phaseolus vulgaris) 
Journal of Experimental Botany  2012;63(8):3109-3125.
Aluminium (Al) toxicity and drought are two major factors limiting common bean (Phaseolus vulgaris) production in the tropics. Short-term effects of Al toxicity and drought stress on root growth in acid, Al-toxic soil were studied, with special emphasis on Al–drought interaction in the root apex. Root elongation was inhibited by both Al and drought. Combined stresses resulted in a more severe inhibition of root elongation than either stress alone. This result was different from the alleviation of Al toxicity by osmotic stress (–0.60 MPa polyethylene glycol) in hydroponics. However, drought reduced the impact of Al on the root tip, as indicated by the reduction of Al-induced callose formation and MATE expression. Combined Al and drought stress enhanced up-regulation of ACCO expression and synthesis of zeatin riboside, reduced drought-enhanced abscisic acid (ABA) concentration, and expression of NCED involved in ABA biosynthesis and the transcription factors bZIP and MYB, thus affecting the regulation of ABA-dependent genes (SUS, PvLEA18, KS-DHN, and LTP) in root tips. The results provide circumstantial evidence that in soil, drought alleviates Al injury, but Al renders the root apex more drought-sensitive, particularly by impacting the gene regulatory network involved in ABA signal transduction and cross-talk with other phytohormones necessary for maintaining root growth under drought.
doi:10.1093/jxb/ers038
PMCID: PMC3350927  PMID: 22371077
Abscisic acid; aluminum; callose; common bean; cytokinin; drought; gene expression; root growth
7.  De novo sequencing and characterization of floral transcriptome in two species of buckwheat (Fagopyrum) 
BMC Genomics  2011;12:30.
Background
Transcriptome sequencing data has become an integral component of modern genetics, genomics and evolutionary biology. However, despite advances in the technologies of DNA sequencing, such data are lacking for many groups of living organisms, in particular, many plant taxa. We present here the results of transcriptome sequencing for two closely related plant species. These species, Fagopyrum esculentum and F. tataricum, belong to the order Caryophyllales - a large group of flowering plants with uncertain evolutionary relationships. F. esculentum (common buckwheat) is also an important food crop. Despite these practical and evolutionary considerations Fagopyrum species have not been the subject of large-scale sequencing projects.
Results
Normalized cDNA corresponding to genes expressed in flowers and inflorescences of F. esculentum and F. tataricum was sequenced using the 454 pyrosequencing technology. This resulted in 267 (for F. esculentum) and 229 (F. tataricum) thousands of reads with average length of 341-349 nucleotides. De novo assembly of the reads produced about 25 thousands of contigs for each species, with 7.5-8.2× coverage. Comparative analysis of two transcriptomes demonstrated their overall similarity but also revealed genes that are presumably differentially expressed. Among them are retrotransposon genes and genes involved in sugar biosynthesis and metabolism. Thirteen single-copy genes were used for phylogenetic analysis; the resulting trees are largely consistent with those inferred from multigenic plastid datasets. The sister relationships of the Caryophyllales and asterids now gained high support from nuclear gene sequences.
Conclusions
454 transcriptome sequencing and de novo assembly was performed for two congeneric flowering plant species, F. esculentum and F. tataricum. As a result, a large set of cDNA sequences that represent orthologs of known plant genes as well as potential new genes was generated.
doi:10.1186/1471-2164-12-30
PMCID: PMC3027159  PMID: 21232141
8.  Alteration of cell-wall porosity is involved in osmotic stress-induced enhancement of aluminium resistance in common bean (Phaseolus vulgaris L.) 
Journal of Experimental Botany  2010;61(12):3245-3258.
Aluminium (Al) toxicity and drought are the two major abiotic stress factors limiting common bean production in the tropics. Using hydroponics, the short-term effects of combined Al toxicity and drought stress on root growth and Al uptake into the root apex were investigated. In the presence of Al stress, PEG 6000 (polyethylene glycol)-induced osmotic (drought) stress led to the amelioration of Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1. PEG 6000 (>> PEG 1000) treatment greatly decreased Al accumulation in the 1 cm root apices even when the roots were physically separated from the PEG solution using dialysis membrane tubes. Upon removal of PEG from the treatment solution, the root tips recovered from osmotic stress and the Al accumulation capacity was quickly restored. The PEG-induced reduction of Al accumulation was not due to a lower phytotoxic Al concentration in the treatment solution, reduced negativity of the root apoplast, or to enhanced citrate exudation. Also cell-wall (CW) material isolated from PEG-treated roots showed a low Al-binding capacity which, however, was restored after destroying the physical structure of the CW. The comparison of the Al3+, La3+, Sr2+, and Rb+ binding capacity of the intact root tips and the isolated CW revealed the specificity of the PEG 6000 effect for Al. This could be due to the higher hydrated ionic radius of Al3+ compared with other cations (Al3+ >> La3+ > Sr2+ > Rb+). In conclusion, the results provide circumstantial evidence that the osmotic stress-inhibited Al accumulation in root apices and thus reduced Al-induced inhibition of root elongation in the Al-sensitive genotype VAX 1 is related to the alteration of CW porosity resulting from PEG 6000-induced dehydration of the root apoplast.
doi:10.1093/jxb/erq146
PMCID: PMC2905193  PMID: 20511277
Aluminium; apoplast; drought stress; intercellular space; organic acids; polyethylene glycol; root elongation
9.  Are Pectins Involved in Cold Acclimation and De-acclimation of Winter Oil-seed Rape Plants? 
Annals of Botany  2008;101(4):521-530.
Background and Aims
The hypothesis was tested that pectin content and methylation degree participate in regulation of cell wall mechanical properties and in this way may affect tissue growth and freezing resistance over the course of plant cold acclimation and de-acclimation.
Methods
Experiments were carried on the leaves of two double-haploid lines of winter oil-seed rape (Brassica napus subsp. oleifera), differing in winter survival and resistance to blackleg fungus (Leptosphaeria maculans).
Key Results
Plant acclimation in the cold (2 °C) brought about retardation of leaf expansion, concomitant with development of freezing resistance. These effects were associated with the increases in leaf tensile stiffness, cell wall and pectin contents, pectin methylesterase (EC 3·1·1·11) activity and the low-methylated pectin content, independently of the genotype studied. However, the cold-induced modifications in the cell wall properties were more pronounced in the leaves of the more pathogen-resistant genotype. De-acclimation promoted leaf expansion and reversed most of the cold-induced effects, with the exception of pectin methylesterase activity.
Conclusions
The results show that the temperature-dependent modifications in pectin content and their methyl esterification degree correlate with changes in tensile strength of a leaf tissue, and in this way affect leaf expansion ability and its resistance to freezing and to fungus pathogens.
doi:10.1093/aob/mcm329
PMCID: PMC2710196  PMID: 18222909
Brassica napus subsp; oleifera; cell wall; cold acclimation; de-acclimation; freezing; growth; leaf stiffness; pathogen; pectins; pectin methylesterase
10.  The Composition of Root Exudates from Two Different Resistant Peanut Cultivars and Their Effects on the Growth of Soil-Borne Pathogen 
The high incidence of various soil-borne diseases in the monoculture field of peanut is a major production constraint in the red soil regions of southern China. The peanut root exudates are generally thought to play an important role in regulating soil-borne pathogens. The responses of the soil-borne pathogens, Fusarium oxysporum and F. solani to the peanut root exudates were studied using one susceptible cultivar Ganhua-5 (GH) and one mid-resistant cultivar Quanhua-7 (QH) as the test materials. The components and contents of the amino acids, sugars and phenolic acids in the peanut root exudates were determined. The results demonstrated that the root exudates from both susceptible and mid-resistant cultivars significantly promoted the spore germination, sporulation and mycelial growth of soil-borne pathogens, F. oxysporum, F. solani compared with the control. The extent of the stimulation was depended on the strains of the Fusarium tested, and gradually increased with the increased concentrations of peanut root exudates. HPLC analysis showed that the contents of sugars, alanine, total amino acids in the root exudates of GH were significantly higher than that in QH, whereas the contents of p-hydroxybenzoic acid, benzoic acid, p-coumaric acid and total phenolic acids were significantly lower than that in QH. Results of the study suggested that the differences in the root exudates from the different peanut cultivars were considered to regulate the wilt-resistance mechanism in the rhizosphere of peanut. The results are therefore crucial important to illustrate the mechanism of peanut replanted obstacle, and to develop its control techniques in the red soil regions of southern China.
doi:10.7150/ijbs.5579
PMCID: PMC3572399  PMID: 23412138
Peanut; Root exudates; Soil-borne pathogens; Amino acids; Phenolic acids.
11.  Phosphorus Enhances Al Resistance in Al-resistant Lespedeza bicolor but not in Al-sensitive L. cuneata Under Relatively High Al Stress 
Annals of Botany  2008;102(5):795-804.
Background and Aims
Aluminium (Al) toxicity and phosphorus (P) deficiency often co-exist in acidic soils and limit crop production worldwide. Lespedeza bicolor is a leguminous forage species that grows very well in infertile, acidic soils. The objective of this study was to investigate the effects of Al and P interactions on growth of Lespedeza and the distributions of Al and P in two different Al-resistant species, and to explore whether P can ameliorate the toxic effect of Al in the two species.
Methods
Two species, Lespedeza bicolor and L. cuneata, were grown for 30 d with alternate Al and P treatments in a hydroponics system. Harvested roots were examined using a root-system scanner, and the contents of Al, P and other nutrient elements in the plants were determined using inductively coupled plasma-atomic emission spectroscopy (ICP-AES). Haematoxylin staining was used to observe the distribution of Al in the roots of seedlings. After pre-culture with or without P application, organic acids in the exudates of roots exposed to Al were held in an anion-exchange resin, eluted with 2 m HCl and then analysed using high-performance liquid chromatography (HPLC).
Key Results
Lespedeza bicolor exhibited a stronger Al resistance than did L. cuneata; Al exclusion mechanisms may mainly be responsible for resistance. P application alleviated the toxic effect of Al on root growth in L. bicolor, while no obvious effects were observed in L. cuneata. Much less Al was accumulated in roots of L. bicolor than in L. cuneata after P application, and the P contents in both roots and shoots increased much more for L. bicolor than for L. cuneata. Lespedeza bicolor showed a higher P/Al ratio in roots and shoots than did L. cuneata. P application decreased the Al accumulation in root tips of L. bicolor but not in L. cuneata. The amount of Al-induced organic acid (citrate and malate) exudation from roots pre-cultured with P was much less than from roots without P application; no malate and citrate exudation was detected in L. cuneata.
Conclusions
P enhanced Al resistance in the Al-resistant L. bicolor species but not in the Al-sensitive L. cuneata under relatively high Al stress, although P in L. cuneata might also possess an alleviative potential. Enhancement of Al resistance by P in the resistant species might be associated with its more efficient P accumulation and translocation to shoots and greater Al exclusion from root tips after P application, but not with an increased exudation of organic acids from roots.
doi:10.1093/aob/mcn166
PMCID: PMC2712389  PMID: 18757448
Lespedeza bicolor; L. cuneata; Al toxicity; Al resistance; root morphology; phosphorus
12.  The lateral root initiation index: an integrative measure of primordium formation 
Annals of Botany  2009;103(5):807-817.
Background and Aims
Lateral root initiation is an essential and continuous process in the formation of root systems; therefore, its quantitative analysis is indispensable. In this study a new measure of lateral root initiation is proposed and analysed, namely the lateral root initiation index (ILRI), which defines how many lateral roots and/or primordia are formed along a parent-root portion corresponding to 100 cortical cells in a file.
Methods
For data collection, a commonly used root clearing procedure was employed, and a new simple root clearing procedure is also proposed. The ILRI was determined as 100dl, where d is the density of lateral root initiation events (number mm−1) and l is the average fully elongated cortical cell length (mm).
Key Results
Analyses of different Arabidopsis thaliana genotypes and of a crop plant, tomato (Solanum lycopersicum), showed that ILRI is a more precise parameter than others commonly used as it normalizes root growth for variations in cell length. Lateral root primordium density varied in the A. thaliana accessions Col, Ler, Ws, and C24; however, in all accessions except Ws, ILRI was similar under the same growth conditions. The nitrogen/carbon ratio in the growth medium did not change the lateral root primordium density but did affect ILRI. The ILRI was also modified in a number of auxin-related mutants, revealing new root branching phenotypes in some of these mutants. The rate of lateral root initiation increased with Arabidopsis seedling age; however, ILRI was not changed in plants between 8 and 14 d post-germination.
Conclusions
The ILRI allows for a more precise comparison of lateral root initiation under different growth conditions, treatments, genotypes and plant species than other comparable methods.
doi:10.1093/aob/mcn267
PMCID: PMC2707874  PMID: 19151042
Arabidopsis thaliana; auxin; lateral root density; lateral root initiation index; mutant phenotype; pericycle; root architecture; root branching; root primordium; Solanum lycopersicum
13.  Aluminum stress signaling in plants 
Plant Signaling & Behavior  2009;4(7):592-597.
Aluminum (Al) toxicity is a major constraint for crop production in acidic soil worldwide. When the soil pH is lower than 5, Al3+ is released to the soil and enters into root tip cell ceases root development of plant. In acid soil with high mineral content, Al is the major cause of phytotoxicity. The target of Al toxicity is the root tip, in which Al exposure causes inhibition of cell elongation and cell division, leading to root stunting accompanied by reduced water and nutrient uptake. A variety of genes have been identified that are induced or repressed upon Al exposure. At tissue level, the distal part of the transition zone is the most sensitive to Al. At cellular and molecular level, many cell components are implicated in the Al toxicity including DNA in nucleus, numerous cytoplastic compounds, mitochondria, the plasma membrane and the cell wall. Although it is difficult to distinguish the primary targets from the secondary effects so far, understanding of the target sites of the Al toxicity is helpful for elucidating the mechanisms by which Al exerts its deleterious effects on root growth. To develop high tolerance against Al stress is the major goal of plant sciences. This review examines our current understanding of the Al signaling with the physiological, genetic and molecular approaches to improve the crop performance under the Al toxicity. New discoveries will open up new avenues of molecular/physiological inquiry that should greatly advance our understanding of Al tolerance mechanisms. Additionally, these breakthroughs will provide new molecular resources for improving the crop Al tolerance via molecular-assisted breeding and biotechnology.
PMCID: PMC2710549  PMID: 19820334
aluminum; toxicity; tolerance; signal transduction; plants
14.  Fagopyrum tataricum (Buckwheat) Improved High-Glucose-Induced Insulin Resistance in Mouse Hepatocytes and Diabetes in Fructose-Rich Diet-Induced Mice 
Experimental Diabetes Research  2012;2012:375673.
Fagopyrum tataricum (buckwheat) is used for the treatment of type 2 diabetes mellitus in Taiwan. This study was to evaluate the antihyperglycemic and anti-insulin resistance effects of 75% ethanol extracts of buckwheat (EEB) in FL83B hepatocytes by high-glucose (33 mM) induction and in C57BL/6 mice by fructose-rich diet (FRD; 60%) induction. The active compounds of EEB (100 μg/mL; 50 mg/kg bw), quercetin (6 μg/mL; 3 mg/kg bw), and rutin (23 μg/mL; 11.5 mg/kg bw) were also employed to treat FL83B hepatocytes and animal. Results indicated that EEB, rutin, and quercetin + rutin significantly improved 2-NBDG uptake via promoting Akt phosphorylation and preventing PPARγ degradation caused by high-glucose induction for 48 h in FL83B hepatocytes. We also found that EEB could elevate hepatic antioxidant enzymes activities to attenuate insulin resistance as well as its antioxidation caused by rutin and quercetin. Finally, EEB also inhibited increases in blood glucose and insulin levels of C57BL/6 mice induced by FRD.
doi:10.1155/2012/375673
PMCID: PMC3324901  PMID: 22548048
15.  Bioactivity comparison of extracts from various parts of common and tartary buckwheats: evaluation of the antioxidant- and angiotensin-converting enzyme inhibitory activities 
Background
Buckwheat flour and buckwheat sprouts possess antioxidant properties, and previous studies have reported on buckwheat flour displaying an inhibitory activity for angiotensin-I converting enzyme (ACE). Information is lacking on the bioactivity of other parts of the buckwheat, such as the seed hulls and plant stalks. This study investigates the ACE inhibitory activity and antioxidant activity of various parts of 2 types of buckwheat, namely, common buckwheat (Fagopyrum esculentum Moench) and tartary buckwheat (Fagopyrum tataricum Gaertn).
Results
The extract of common hulls extracted using 50% (v/v)-ethanol solvent presented a remarkable inhibitory activity. The value of IC50 is 30 μg ml-1. The extracts of both common and tartary hulls extracted using 50% (v/v)-ethanol solvent demonstrated an antioxidant activity that is superior to that of other extracts.
Conclusion
This study determined that the ethanolic extract of the hulls of common buckwheat presented more favorable antioxidant and ACE inhibitory abilities. However, the correlation of antioxidant activity and ACE inhibitory activity for all 18 types of extracts is low. The ACE inhibitory activity could have been caused by a synergistic effect of flavonoids or from other unidentified components in the extracts. The ethanolic extract of common hulls demonstrated remarkable ACE inhibitory activity and is worthy of further animal study.
doi:10.1186/1752-153X-6-78
PMCID: PMC3485629  PMID: 22853321
Tartary buckwheat; Common buckwheat; ACE inhibition; Microplate fluometric assay
16.  Identification of Sources of Resistance to Four Species of Root-knot Nematodes in Tobacco 
Journal of Nematology  1999;31(3):272-282.
Resistance to the southern root-knot nematode, Meloidogyne incognita races 1 and 3, has been identified, incorporated, and deployed into commercial cultivars of tobacco, Nicotiana tabacum. Cultivars with resistance to other economically important root-knot nematode species attacking tobacco, M. arenaria, M. hapla, M. javanica, and other host-specific races of M. incognita, are not available in the United States. Twenty-eight tobacco genotypes of diverse origin and two standard cultivars, NC 2326 (susceptible) and Speight G 28 (resistant to M. incognita races 1 and 3), were screened for resistance to eight root-knot nematode populations of North Carolina origin. Based on root gall indices at 8 to 12 weeks after inoculation, all genotypes except NC 2326 and Okinawa were resistant to M. arenaria race 1, and races 1 and 3 of M. incognita. Except for slight root galling, genotypes resistant to M. arenaria race 1 responded similarly to races 1 and 3 of M. incognita. All genotypes except NC 2326, Okinawa, and Speight G 28 showed resistance to M. javanica. Okinawa, while supporting lower reproduction of M. javanica than NC 2326, was rated as moderately susceptible. Tobacco breeding lines 81-R-617A, 81-RL- 2K, SA 1213, SA 1214, SA 1223, and SA 1224 were resistant to M. arenaria race 2, and thus may be used as sources of resistance to this pathogen. No resistance to M. hapla and only moderate resistance to races 2 and 4 of M. incognita were found in any of the tobacco genotypes. Under natural field infestations of M. arenaria race 2, nematode development on resistant tobacco breeding lines 81-RL-2K, SA 1214, and SA 1215 was similar to a susceptible cultivar with some nematicide treatments; however, quantity and quality of yield were inferior compared to K 326 plus nematicides.
PMCID: PMC2620380  PMID: 19270897
Javanese root-knot nematode; Meloidogyne species; nematode; resistance; southern root-knot nematode; tobacco
17.  Early Root Response to Meloidogyne incognita in Resistant and Susceptible Alfalfa Cultivars 
Journal of Nematology  1996;28(4):475-484.
The early events of Meloidogyne incognita behavior and associated host responses following root penetration were studied in resistant (cv. Moapa 69) and susceptible (cv. Lahontan) alfalfa. Ten-day-old seedlings of alfalfa cultivars were inoculated with second-stage juveniles (J2) and harvested 12, 24, 48, and 72 hours and 7, 14, and 21 days later. Both cultivars supported similar root penetration and initial J2 migration. By 72 hours after inoculation the majority of J2 were amassed inside the vascular cylinder in roots of susceptible Lahontan, while J2 had not entered the vascular cylinder of resistant Moapa 69 and remained clumped at the root apex. Nematode development progressed normally in Lahontan, but J2 were not observed in Moapa 69 after day 7. The greatest differences between RNA translation products isolated from inoculated and uninoculated roots of Lahanton occurred 72 hours after inoculation. Only minor differences in gene expression were observed between inoculated and uninoculated Moapa 69 roots at 72 hours. Comparison of translation products from inoculated versus mechanically wounded Lahontan roots revealed products that were specific to or enhanced in nematode-infected plants. Moapa 69 appears to possess a type of resistance to M. incognita that does not depend on a conventional hypersensitive response.
PMCID: PMC2619717  PMID: 19277165
alfalfa; gene expression; host response; Lahontan; Medicago sativa; Meloidogyne incognita; Moapa 69; resistance; RNA; root-knot nematode
18.  Effect of Field Crops on Population Densities of Pratylenchus neglectus and P. thornei in Southeastern Australia; Part 1: P. neglectus 
Journal of Nematology  2000;32(4S):591-599.
Eighty-one cultivars from 12 field crop species were assessed for suitability as hosts to the root lesion nematode, Pratylenchus neglectus, in two field trials. Host status was assessed on the basis of either final P. neglectus densities in soil or multiplication rate under different crops. Both techniques gave consistent results for crop and cultivar ranking, and it was therefore concluded that, in these trials, final population density could be used for screening cultivars for resistance to P. neglectus. Differences were observed among crops and cultivars for host suitability to P. neglectus. Chickpea, wheat, and canola were good hosts, while barley, oat, durum wheat, medic, and vetch were moderate hosts. Field pea, faba bean, and triticale were poor hosts. A range in host suitability was observed for wheat, barley, and oat cultivars.
PMCID: PMC2620495  PMID: 19271014
barley; canola; chickpea; control; faba bean; field pea; host suitability; medic; multiplication; oat; resistance; root lesion nematode; rotation; susceptibility; triticale; vetch; wheat
19.  Root-to-shoot Cd translocation via the xylem is the major process determining shoot and grain cadmium accumulation in rice 
Journal of Experimental Botany  2009;60(9):2677-2688.
Physiological properties involved in divergent cadmium (Cd) accumulation among rice genotypes were characterized using the indica cultivar ‘Habataki’ (high Cd in grains) and the japonica cultivar ‘Sasanishiki’ (low Cd in grains). Time-dependence and concentration-dependence of symplastic Cd absorption in roots were revealed not to be responsible for the different Cd accumulation between the two cultivars because root Cd uptake was not greater in the Cd-accumulating cultivar ‘Habataki’ compared with ‘Sasanishiki’. On the other hand, rapid and greater root-to-shoot Cd translocation was observed in ‘Habataki’, which could be mediated by higher abilities in xylem loading of Cd and transpiration rate as a driving force. To verify whether different abilities in xylem-mediated shoot-to-root translocation generally account for the genotypic variation in shoot Cd accumulation in rice, the world rice core collection, consisting of 69 accessions which covers the genetic diversity of almost 32 000 accessions of cultivated rice, was used. The results showed strong correlation between Cd levels in xylem sap and shoots and grains among the 69 rice accessions. Overall, the results presented in this study revealed that the root-to-shoot Cd translocation via the xylem is the major and common physiological process determining the Cd accumulation level in shoots and grains of rice plants.
doi:10.1093/jxb/erp119
PMCID: PMC2692013  PMID: 19401409
Cadmium; genotypic variation; Oryza sativa; uptake; xylem loading
20.  A role for redox factors in shaping root architecture under phosphorus deficiency 
Plant Signaling & Behavior  2010;5(1):64-66.
The developmental response of the Arabidopsis root system to low phosphorus (P) availability involves the reduction in primary root elongation accompanied by the formation of numerous lateral roots. We studied the roles of selected redox metabolites, namely, radical oxygen species (ROS) and ascorbic acid (ASC) in the regulation of root system architecture by different P availability. Rapidly growing roots of plants grown on P-sufficient medium synthesize ROS in root elongation zone and quiescent centre. We have demonstrated that the arrest of root elongation at low P medium coincides with the disappearance of ROS from the elongation zone. P-starvation resulted in a decrease in ascorbic acid level in roots. This correlated with a decrease in cell division activity. On the other hand, feeding P-deficient plants with ASC, stimulated mitotic activity in the primary root meristem and partly reversed the inhibition of root growth imposed by low P conditions. In this paper, we discuss the idea of the involvement of redox agents in the regulation of root system architecture under low P availability.
PMCID: PMC2835962  PMID: 20592813
ascorbic acid; phosphate deficiency; primary root; radical oxygen species; root growth; root system architecture
21.  Analysis of LuPME3, a pectin methylesterase from Linum usitatissimum, revealed a variability in PME proteolytic maturation 
Plant Signaling & Behavior  2012;7(1):59-61.
Pectin methylesterase (PME) catalyzes the de-methylesterification of pectin in plant cell walls during cell elongation.1 Pectins are mainly composed of α(1, 4)-D-galacturonosyl acid units that are synthesized in a methylesterified form in the Golgi apparatus to prevent any interaction with Ca2+ ions during their intracellular transport.2 The highly methylesterified pectins are then secreted into the apoplasm3 and subsequently de-methylesterified in muro by PMEs. This can either induce the formation of pectin gels through the Ca2+ crosslinking of neighboring non-methylesterified chains or create substrates for pectin-degrading enzymes such as polygalacturonases and pectate lyases for the initiation of cell wall loosening.4 PMEs belong to a large multigene family. Sixty­six PME-related genes are predicted in the Arabidopsis genome.1 Among them, we have recently shown that AtPME3 (At3g14310), a major basic PME isoform in A. thaliana, is ubiquitously expressed in vascular tissues and play a role in adventitious rooting.5 In flax (Linum usitatissimum), three genes encoding PMEs have been sequenced so far, including LuPME3, the ortholog of AtPME3. Analysis of the LuPME3 isoform brings new insights into the processing of these proteins.
doi:10.4161/psb.7.1.18632
PMCID: PMC3357370  PMID: 22301970
Linum usitatissimum; cell wall; pectin; pectin methylesterase; protein maturation
22.  Evolution of root plasticity responses to variation in soil nutrient distribution and concentration 
Evolutionary Applications  2012;5(8):850-857.
Root plasticity, a trait that can respond to selective pressure, may help plants forage for nutrients in heterogeneous soils. Agricultural breeding programs have artificially selected for increased yield under comparatively homogeneous soil conditions, potentially decreasing the capacity for plasticity in crop plants like barley (Hordeum vulgare). However, the effects of domestication on the evolution of root plasticity are essentially unknown. Using a split container approach, we examined the differences in root plasticity among three domestication levels of barley germplasm (wild, landrace, and cultivar) grown under different concentrations and distribution patterns of soil nutrients. Domestication level, nutrient concentration, and nutrient distribution interactively affected average root diameter; differential root allocation (within-plant plasticity) was greatest in wild barley (Hordeum spontaneum), especially under low nutrient levels. Correlations of within-plant root plasticity and plant size were most pronounced in modern cultivars under low-nutrient conditions. Barley plants invested more resources to root systems when grown in low-nutrient soils and allocated more roots to higher-nutrient locations. Root plasticity in barley is scale dependent and varies with domestication level. Although wild barley harbors a greater capacity for within-plant root plasticity than domesticated barley, cultivars exhibited the greatest capacity to translate within-plant plasticity into increased plant size.
doi:10.1111/j.1752-4571.2012.00263.x
PMCID: PMC3552402  PMID: 23346229
artificial selection; barley; evolution of plasticity; Hordeum spontaneum; Hordeum vulgare; plant domestication
23.  Role of the iron transporter OsNRAMP1 in cadmium uptake and accumulation in rice 
Plant Signaling & Behavior  2011;6(11):1813-1816.
The heavy metal cadmium (Cd) is toxic to humans, and its accumulation in rice grains is a major agricultural problem. Rice has seven putative metal transporter NRAMP genes, but microarray analysis showed that only OsNRAMP1 is highly up-regulated by iron (Fe) deficiency. OsNRAMP1 localized to the plasma membrane and transported Cd as well as Fe. OsNRAMP1 expression was observed mainly in roots and was higher in the roots of a high-Cd-accumulating cultivar (Habataki) than in those of a low-Cd-accumulating cultivar (Sasanishiki). The amino acid sequence of OsNRAMP1 in the Sasanishiki and Habataki cultivars was found to be 100% identical. These results suggest that OsNRAMP1 participates in cellular Cd uptake and that the differences observed in Cd accumulation among cultivars are because of differences in OsNRAMP1 expression levels in roots.
doi:10.4161/psb.6.11.17587
PMCID: PMC3329356  PMID: 22067109
cadmium; metal transporter; OsNRAMP1; phytoremediation; rice
24.  Strigolactones as Germination Stimulants for Root Parasitic Plants 
Plant and Cell Physiology  2010;51(7):1095-1103.
Witchweeds (Striga spp.) and broomrapes (Orobanche and Phelipanche spp.) are the two most devastating root parasitic plants belonging to the family Orobanchaceae and are causing enormous crop losses throughout the world. Seeds of these root parasites will not germinate unless they are exposed to chemical stimuli, ‘germination stimulants’ produced by and released from plant roots. Most of the germination stimulants identified so far are strigolactones (SLs), which also function as host recognition signals for arbuscular mycorrhizal fungi and a novel class of plant hormones inhibiting shoot branching. In this review, we focus on SLs as germination stimulants for root parasitic plants. In addition, we discuss how quantitative and qualitative differences in SL exudation among sorghum cultivars influence their susceptibility to Striga.
doi:10.1093/pcp/pcq055
PMCID: PMC2900819  PMID: 20403809
Germination stimulant; Orobanche; Phelipanche; Root parasitic plants; Striga; Strigolactone
25.  Aluminium-induced inhibition of root elongation in Arabidopsis is mediated by ethylene and auxin 
Journal of Experimental Botany  2009;61(2):347-356.
Aluminium (Al) is phytotoxic when solubilized into Al3+ in acidic soils. One of the earliest and distinct symptoms of Al3+ toxicity is inhibition of root elongation. To decipher the mechanism by which Al3+ inhibits root elongation, the role of ethylene and auxin in Al3+-induced inhibition of root elongation in Arabidopsis thaliana was investigated using the wild type and mutants defective in ethylene signalling (etr1-3 and ein2-1) and auxin polar transport (aux1-7 and pin2). Exposure of wild-type Arabidopsis to AlCl3 led to a marked inhibition of root elongation, and elicited a rapid ethylene evolution and enhanced activity of the ethylene reporter EBS:GUS in root apices. Root elongation in etr1-3 and ein2-1 mutants was less inhibited by Al3+ than that in wild-type plants. Ethylene synthesis inhibitors, Co2+ and aminoethoxyvinylglycine (AVG), and an antagonist of ethylene perception (Ag+) abolished the Al3+-induced inhibition of root elongation. There was less inhibition of root elongation by Al3+ in aux1-7 and pin2 mutants than in the wild type. The auxin polar transport inhibitor, naphthylphthalamic acid (NPA), substantially alleviated the Al3+-induced inhibition of root elongation. The Al3+ and ethylene synthesis precursor aminocyclopropane carboxylic acid (ACC) increased auxin reporter DR5:GUS activity in roots. The Al3+-induced increase in DR5:GUS activity was reduced by AVG, while the Al3+-induced increase in EBS:GUS activity was not altered by NPA. Al3+ and ACC increased transcripts of AUX1 and PIN2, and this effect was no longer observed in the presence of AVG and Co2+. These findings indicate that Al3+-induced ethylene production is likely to act as a signal to alter auxin distribution in roots by disrupting AUX1- and PIN2-mediated auxin polar transport, leading to arrest of root elongation.
doi:10.1093/jxb/erp306
PMCID: PMC2803203  PMID: 19858117
Aluminium toxicity; Arabidopsis thaliana; auxin polar transport; ethylene; root elongation

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