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PubMed2003

[Change of superficial shape and cell microstructure of milk-vetch (Astragalus L.) root under simulated Cu pollution in a red soil].

This study carried out experiments to investigate changes of shape, inter-structures and cell microstructures of milk-vetch root under simulated Cu stress at non- or contaminated levels in a red soil using observation and bio-microscopic technique. It resulted that when Cu concentration ranged from 0 to 40 mg.kg-1 soil, the milk-vetch grew well and it had a whole root which worked normally. When Cu concentration reached 50 mg.kg-1 soil, the growth of milk-vetch began to get influence with decline of biomass, the taproot crooked and was less branched, root became short and hazel and had fewer shorter hairs, tubby appeared, epidermis began to shrink and cell wall cockled slightly and unevenly, the boundary between plasmalemma and organelle blurred as well. When treatment concentration reaching to 200 mg.kg-1 soil, milk-vetch roots became rotted and black, the cell wall broke and cytoplasm shrank so severely that plasmolysis happened and the plant died. So the critical Cu concentration in experimented soil was 50 mg.kg-1 soil, and the resistance of milk-vetch root to Cu contamination buildup with the growth of aboveground part, and cell wall was the main part to Cu tolerance.

Astragalus Plant↗
PubMed2005

[Tolerance of Arundo donax to heavy metals].

This paper studied the tolerance of Arundo donax grown in a simulated heavy metals polluted wetland, and determined the biological characters and chlorophyll contents of the plant at its different growth stages as well as the changes of soil heavy metals contents. The results showed that Arundo donax could survive in the wetland when the concentrations of Cu2+, Pb2+, Cd2+, Zn2+, Ni2+ and Hg2+ were 100 mg x kg(-1) and Cr6+ concentration was 50 mg x kg(-1). During 40 days growth period, the chlorophyll content decreased by 20%-56% and the leaf became soft with its tip withered, but the plant still grew. Compared with control, Arundo donax in the polluted wetland was slight and yellow-green, but the impact on plant height was inconspicuous. Arundo donax treated with 100 mg x kg(-1) Cr6+ grew slowly with its root stock rotted, and its leaves withered in a short time, indicating that the plant could not tolerate the pollution of high concentration Cr6+. The concentrations of soil heavy metals declined with the growth of the plant, probably due to the translocation of heavy metals from peripheral soil to rhizosphere and the phytoextraction and phytovolatilization, because the heavy metals contents in rhizosphere were much higher than those in the bulk soil in the test jar. The characters of large biomass, exuberant root and good adaptability of Arundo donax suggested its great potential in remediation of polluted soils. The study on the application of Arundo donax to phytoremediation is of realistic significance.

Biodegradation, Environmental↗
PubMed1996

Greenhouse spinach production in a NFT system.

Primed spinach (Spinacia oleracea L., cv. Nordic) seed was started in rockwool slabs in a growth room for eight days before the seedlings were transplanted into a controlled environment greenhouse equipped with five identical, but separate, NFT systems. The day and night temperatures in the greenhouse were maintained at 24 and 18 degrees C, respectively, with the daytime starting at 06:00 and ending at 22:00 hr. A photoperiod of 16 hrs was maintained, to prevent early bolting, and different target daily integrated light levels (PPF, in mol m-2 d-1) were studied to observe dry weight production. HPS lamps were used as the supplemental light source. Thirty-three days after seeding a final harvest was performed. Using the expolinear growth equation, dry weight production can be predicted based solely on target daily integrated light levels. Total chlorine residuals in the nutrient solution higher than 1 ppm were observed to be toxic. Root disease (rot) in the plant crown was found to be caused by Fusarium. Several remedies, including three biofungicides and potassium silicate, were tried but none proved to be consistently successful.

Biomass↗
PubMed1979

Race-specific molecules that protect soybeans from Phytophthora megasperma var. sojae.

Phytophthora megasperma var. sojae (A. A. Hildebrand) is a fungal stem and root rot-causing pathogen of soybeans. Glycoproteins secreted into the medium of the aseptically cultured fungus have been partially purified by (NH(4))(2)SO(4) precipitation and by column chromatography on norleucine-substituted Sepharose 4B and on DEAE-cellulose. Glycoprotein preparations from P. megasperma var. sojae races 1, 2, and 3 have been tested on four cultivars of soybeans. The partially purified glycoproteins from incompatible races of the pathogen (races that cannot successfully infect the plant), but not those from compatible races (races that can kill the plant), protect soybean seedlings from attack by compatible races. The seedlings are protected by introducing the glycoproteins into hypocotyl wounds of seedlings either 90 min prior to or at the time of inoculation of the wounds with mycelia of one of the pathogens. The glycoprotein preparations are poor nonspecific elicitors of phytoalexin accumulation; the glycoproteins have less than 1.0% of the elicitor activity of the glucans present in the mycelial walls of the pathogen.

Journal Article↗
PubMed2006

Clonality in South African isolates and evidence for a European origin of the root pathogen Thielaviopsis basicola.

Thielaviopsis basicola is a soil-borne fungal pathogen with a wide host range and a cosmopolitan distribution. It causes disease on many agricultural crops, and in South Africa is the causal agent of black pod rot of groundnuts and black root rot on chicory. Knowledge of the population diversity of T. basicola could provide valuable information regarding management strategies, the possible movement, origin, and reproductive strategies of the fungus. The objective of this study was to determine the population diversity of T. basicola isolates from groundnuts and chicory in South Africa using co-dominant polymorphic markers. These markers were also used to compare isolates from South Africa with those from other hosts and geographic regions. Seven loci revealed nine alleles and two genotypes, one on groundnut and one on chicory, differing at only two loci. T. basicola isolates from eight different countries and ten different hosts revealed 17 genotypes across the seven loci with 39 different alleles. The lack of diversity for the two South African host-related populations of isolates suggests that T. basicola was introduced into South Africa. Some evidence is provided for a European origin of the pathogen, possibly linked to trade in root crops.

Europe↗
PubMed2002

Inability to find consistent bacterial biocontrol agents of Pythium aphanidermatum in cucumber using screens based on ecophysiological traits.

A collection of 821 rhizobacteria from cucumber, originating from different root locations and stages of plant development, was screened for potential biocontrol agents of Pythium aphanidermatum (Edson) Fitzp. The screening procedure exploited carbon source utilization profiles and growth rates of bacteria as indicators of a partial niche overlap with the pathogen. The bacteria were tested for growth on nine carbon sources (glucose, fucose, sucrose, maltose, asparagine, alanine, galacturonic acid, succinic acid, and linoleic acid), most of which are reported to be used by the zoospores of P. aphanidermatum in the infection process. The isolates were classified as fast- or slow-growing, depending on their growth rate in 1/10 strength TSB. By nonhierarchical cluster analysis, 20 clusters were generated of bacteria with similar profiles of carbon source utilization. Redundancy analysis showed that the type of root sample explained 47% of the variance found in the relative abundance of bacteria from the clusters. Bacteria from clusters using none or few of the carbon sources, e.g., maltose and linoleic acid, with many slow-growing isolates, showed a preference for plants in the vegetative or generative stage, or for old root regions (root base). Bacteria from clusters with fast-growing isolates, using many carbon sources, were relatively abundant in the seedling stage. A selection of 127 bacteria from the different clusters was tested for disease suppressive capabilities in bioassays on young cucumber plants in nutrient solution, inoculated with zoospores of P. aphanidermatum. Nine of these bacteria produced biosurfactants, and 27 showed antibiosis against mycelial growth in plate assays. For 31 isolates, significant positive effects on plant biomass were shown, as analyzed with a general linear regression model. For most isolates, these effects occurred only in one of two replicate assays and no reductions in the degree of root and crown rot were found. Of the isolates that used many of the tested carbon sources, only four had positive effects on plant biomass. The majority of the isolates that positively affected plant biomass used few to moderate numbers of carbon sources and did not produce antibiotics or biosurfactants. In conclusion, competition for the tested carbon sources with the zoospores did not play a decisive role in disease suppression, and no clear relation was found between ecophysiological traits and disease suppression. Only isolate 3.1T8, isolated from root tips in the generative stage of plant growth, significantly increased plant biomass and suppressed root and crown rot symptoms in five out of six bioassays. The isolate produced an antifungal substance in plate assays and showed biosurfactant production in several (cucumber-derived) media.

Bacteria↗
PubMed1987

Growth and disease response of soybeans from early maturity groups to ozone and Fusarium oxysporum.

The single and combined effects of ozone (O(3)) and Fusarium oxysporum on growth and disease expression of soybean genotypes differing in foliar sensitivity to O(3) were studied in the greenhouse. O(3) had no effect on root and hypocotyl rot severity of PI 153.283 (O(3)-sensitive, S) or PI 189.907 (O(3)-tolerant, T) maturity group I soybean lines. Plants of both genotypes infected with F. oxysporum and exposed to O(3) had greater reductions in relative growth rate (RGR), net assimilation rate (NAR), and had more stippled leaves per plant than Fusarium-free plants exposed to O(3). O(3) alone had a greater impact on shoot dry weight, RGR, and NAR of PI 153.283 (S) than of PI 189.907 (T). O(3) alone reduced shoot and root dry weights primarily through a depression in NAR and less through reduced leaf area. F. oxysporum alone reduced root dry weight at 35 days; however, infected plants responded with increases in root dry weight from 49 to 63 days. Similarly, F. oxysporum alone lowered early RGR but subsequent RGR decline was less rapid while NAR remained high, particularly during later sampling intervals. Infection by F. oxysporum that causes root and hypocotyl rot increased soybean sensitivity to O(3) by prolonging active vegetative growth.

Journal Article↗
PubMed1982

Studies on a Factor in Sweet Potato Root Which Agglutinates Spores of Ceratocystis fimbriata, Black Rot Fungus.

A factor which agglutinated the spores of Ceratocystis fimbriata in the presence of Ca(2+) was purified from sweet potato (Ipomea batatas Lam cv. Norin[1]) root. Element composition of the purified factor was as follows; analysis found: C (29.8%), H (3.97%), O (65.34%), N (0.81%): calculated for C(43)H(69)O(70)N(1): C (30.02%), H (4.01%), O (65.15%), N (0.81%). The factor was mainly composed of galacturonic acid (53% of dry weight) and contained arabinose, fucose, and unidentified component as minor components. The factor also agglutinated A-, B-, AB-, and O types of human erythrocytes to almost the same degree in the presence of Ca(2+). The differential spore-agglutinating activity of the factor depended on the pH of the assay medium; it agglutinated similarly the germinated spores of sweet potato and coffee strains at pH 7.5 and 5.5, whereas it displayed a distinct differential agglutinating activity at pH 6.5. The factor was assayed for spore-agglutinating activity at pH 6.5, using the germinated and ungerminated spores of seven strains of C. fimbriata; sweet potato, coffee, prune, cacao, oak, taro, and almond strains. The factor agglutinated ungerminated spores of all seven strains similarly, although small differences were observed among strains. On the other hand, a clear differential agglutination was observed among the germinated spores of various strains; sweet potato and almond strains were highly insensitive in comparison with other strains. The growth of the agglutinated spores of C. fimbriata was inhibited. These results are discussed in relation to host-parasite specificity.

Journal Article↗
PubMed2003

Analysis of rDNA ITS sequences to determine genetic relationships among, and provide a basis for simplified diagnosis of, Fusarium species causing crown rot and head blight of cereals.

Genetic relationships of the complex of Fusarium species associated with crown rot and head blight in cereals and some species associated with plant diseases in general were examined by distance and maximum parsimony algorithms of their internal transcribed spacer sequences. The analysis clustered the complex of Fusarium species that causes root and crown rot and head blight of cereals and three other clusters of F. sambucinum, F. venenatum and F. poae into one clade. This group of Fustarium species was also found in this study to correspond to the group defined by the presence of the tri5 gene. The tri5 gene was recently reported to co-segregate with the locus governing the type of trichothecene produced, and probably maps in the trichothecene gene cluster. The other clusters of F. avenaceum, F. tricinctum, F. torulosum, F. oxysporum, F. verticillioides and F. solani did not have the tri5 gene. Although, F. pseudograminearum was phylogenetically close with the cluster of F. graminearum, F. culmorum and F. cerealis, it could be distinctly separated from them. The distinct genetic status of F. pseudograminearum from F. graminearum corroborated with other published molecular data and isozyme findings. The molecular analysis provided a simple diagnostic tool to differentiate fungi causing crown rot from those involved in head blight.

Cluster Analysis↗
PubMed1978

Isolation and Characterization of Factors in Sweet Potato Root Which Agglutinate Germinated Spores of Ceratocystis fimbriata, Black Rot Fungus.

A factor which agglutinates the germinated spores of Ceratocystis fimbriata was isolated from the sweet potato root. The factor is a glycoprotein with a molecular weight of 1.6 x 10(6) daltons and required divalent cations such as Ca(2+), Mn(2+), Ni(2+), and Mg(2+) for activity. The activity of the factor was pH-dependent. The factor also agglutinated rabbit erythrocytes and is classified as a phytohemagglutinin or lectin. The factor agglutinated germinated spores of seven strains of C. fimbriata to almost the same degree. The factor showed differential agglutinating activity toward the strains in the presence of unidentified low molecular weight factor(s) in the sweet potato root. These results support our earlier suggestion that the spore-agglutinating factors in host plants function as the determinants of specificity in some host-parasite interactions.

Journal Article↗
PubMed1987

Pathotoxin effects in sorghum are also produced by mercuric chloride treatment.

Pathogenic isolates of Periconia circinata produce a host-specific toxin (PC-toxin) and cause a root and crown rot in susceptible genotypes of sorghum. Treatment with PC-toxin leads to selective development of disease symptoms and an increase in synthesis of a group of acidic, low molecular weight proteins only in susceptible genotypes. Treatment of sorghum seedlings or excised root tips with HgCl(2) resulted in responses indistinguishable from those produced by treatment with PC-toxin, but the effects were not genotype specific.

Journal Article↗
PubMed2005

Fumonisin B1, a sphingoid toxin, is a potent inhibitor of the plasma membrane H+-ATPase.

Fumonisin B(1) (FB(1)) is an amphipathic toxin produced by the pathogenic fungus Fusarium verticillioides which causes stem, root and ear rot in maize (Zea mays L.). In this work, we studied the action of FB(1) on the plasma membrane H(+)-ATPase (EC 3.6.1.34) from germinating maize embryos, and on the fluidity and lipid peroxidation of these membranes. In maize embryos the toxin at 40 microM inhibited root elongation by 50% and at 30 microM decreased medium acidification by about 80%. Irrespective of the presence and absence of FB(1), the H(+)-ATPase in plasma membrane vesicles exhibited non-hyperbolic saturation kinetics by ATPH-Mg, with Hill number of 0.67. Initial velocity studies revealed that FB(1) is a total uncompetitive inhibitor of this enzyme with an inhibition constant value of 17.5+/-1 microM. Thus FB(1) decreased V(max) and increased the apparent affinity of the enzyme for ATP-Mg to the same extent. Although FB(1) increased the fluidity at the hydrophobic region of the membrane, no correlation was found with its effect on enzyme activity, since both effects showed different FB(1)-concentration dependence. Peroxidation of membrane lipids was not affected by the toxin. Our results suggest that, under in vivo conditions, the plasma membrane H(+)-ATPase is a potentially important target of the toxin, as it is inhibited not only by FB(1) but also by its structural analogs, the sphingoid intermediates, which accumulate upon the inhibition of sphinganine N-acyltransferase by this toxin.

Cell Membrane↗
PubMed2001

Biological activity of Pythium oligandrum against Phytophthora species.

Influence of Pythium oligandrum as an a.i. of Polyversum on population dynamic of Phytophthora cryptogea in peat and development of Phytophtora rot on gerbera, Lawson cypress and yew-tree were evaluated. Drenching of peat, artificially infested with P. cryptogea, with Polyversum immediatelly after gerbera planting resulted in significant decrease of colony forming units number within 4 weeks. Concentrations of the biopreparate used had no significant influence on its greater, biological activity. Drenching of plants with Polyversum at conc. 0.05 or 0.1%, after planting into peat infested with P. cryptogea or P. cinnamomi, resulted in the strong suppression of Phytophthora foot or root and stem rot of gerbera, cypress and yew-tree.

Antibiosis↗
PubMed1996

The Phytophthora sojae genome contains tandem repeat sequences which vary from strain to strain.

The oömycete Phytophthora sojae causes root and stem rot of soybean. In P. sojae, 37 cultivarspecific physiological races have been reported with different reactions against 13 single dominant resistance genes in soybean. Recent genetic studies have demonstrated that the P. sojae-soybean interaction follows Flor's gene-for-gene model. Genomic subtraction was carried out to isolate race-specific DNA sequences possibly including avirulence genes. DNA from a race 1 isolate was subtraction-enriched with the DNAs from race 19 and race 22 isolates. The enriched DNAs were used to differentially screen a P. sojae genomic library. Characterization of the clones obtained identified repetitive sequences with variable copy numbers among isolates. Five of these repetitive sequences were tandemly repeated and were localized on single chromosomes. The sixth corresponded to the ribosomal RNA genes. These observations suggest that gene amplification could contribute to the generation of genetic diversity in P. sojae. Enriched sequences corresponding to avirulence genes were not detected.

Base Sequence↗
PubMed2005

Identification of a large cluster of coiled coil-nucleotide binding site--leucine rich repeat-type genes from the Rps1 region containing Phytophthora resistance genes in soybean.

Fifteen Rps genes confer resistance against the oomycete pathogen Phytophthora sojae, which causes root and stem rot disease in soybean. We have isolated a disease resistance gene-like sequence from the genomic region containing Rps1-k. Four classes of cDNA of the sequence were isolated from etiolated hypocotyl tissues that express the Rps1-k-encoded Phytophthora resistance. Sequence analyses of a cDNA clone showed that the sequence is a member of the coiled coil-nucleotide binding site-leucine rich repeat (CC-NBS-LRR)-type of disease resistance genes. It showed 36% identity to the recently cloned soybean resistance gene Rpg1-b, which confers resistance against Pseudomonas syringae pv. glycinea, and 56% and 38% sequence identity to putative resistance gene sequences from lotus and Medicago truncatula, respectively. The soybean genome contains about 38 copies of the sequence. Most of these copies are clustered in approximately 600 kb of contiguous DNA of the Rps1-k region. We have identified a recombinant that carries both rps1-k- and Rps1-k-haplotype-specific allelomorphs of two Rps1-k-linked molecular markers. An unequal crossover event presumably led to duplication of alleles for these two physically linked molecular markers. We hypothesize that the unequal crossing over was one of the mechanisms involved in tandem duplication of CC-NBS-LRR sequences in the Rps1-k region.

Amino Acid Sequence↗
PubMed2001

Cloning and expression analysis of NhL1, a gene encoding an extracellular lipase from the fungal pea pathogen Nectria haematococca MP VI (Fusarium solani f. sp. pisi) that is expressed in planta.

The filamentous fungus Nectria haematococca (anamorph Fusarium solani f. sp. pisi) resides in soil, and attacks pea seedlings in the area of the underground epicotyl and upper tap root, causing foot rot disease. We detected lipase activity during in vitro growth of N. haematococca. Subsequently, a lipase gene was cloned and functionally characterised by heterologous expression in Saccharomyces cerevisiae. The full-length cDNA of 1152 bp was cloned using a 3' RACE-PCR approach coupled with cDNA library screening. The genomic clone, comprising an ORF of 999 bp interrupted by two introns of 56 and 64 bp, was isolated from a newly constructed lambda phage library. Analysis of the deduced protein sequence revealed the presence of a typical signal peptide at the N-terminus, and of the three conserved amino acids forming the active site of lipases. The lipase of N. haematococca has a low degree of similarity to the lipases from Humicola lanuginosa (37.2%), Rhizomucor miehei (21.6%), Rhizopus delemar (23.1%), Rhizopus niveus (25.9%), and to mono- and diacylglycerol lipase from Penicillium camembertii (30.8%), and very high similarity (94.6%) to a lipase from Fusarium heterosporum. The lipase from N. haematococca shows maximal activity at 37 degrees C and pH 8.0. Based on Southern analysis, the lipase clone represents a single-copy gene in N. haematococca. Expression analysis was performed by RT-PCR. In vitro, the lipase gene shows a low basal expression, but is highly inducible by lipase substrates, and repressed by glucose. During plant infection, transcripts of this fungal lipase gene were detected 4, 8, and 10 days after infection.

Amino Acid Sequence↗
PubMed2004

Phytophthora sojae avirulence genes Avr4 and Avr6 are located in a 24kb, recombination-rich region of genomic DNA.

A cross between two different races (race 7xrace 25) of the soybean root and stem rot pathogen Phytophthora sojae was analyzed to characterize the genomic region flanking two cosegregating avirulence genes, Avr4 and Avr6. Both genes cosegregated in the ratio of 82:17 (avirulent:virulent) in an F(2) population, suggestive of a single locus controlling both phenotypes. A chromosome walk was commenced from RAPD marker OPE7.1C, 2.0cM distant from the Avr4/6 locus. Three overlapping cosmids were isolated which included genetic markers that flank the Avr4/6 locus. The chromosome walk spanned a physical distance of 67kb which represented a genetic map distance of 22.3cM, an average recombination frequency of 3.0kb/cM and 11.7-fold greater than the predicted average recombination frequency of 35.3kb/cM for the entire P. sojae genome. Six genes (cDNA clones) expressed from the Avr4/6 genomic region encompassed by the cosmid contig were identified. Single nucleotide polymorphisms and restriction fragment length polymorphisms showed these six genes were closely linked to the Avr4/6 locus. Physical mapping of the cDNA clones within the cosmid contig made it possible to deduce the precise linkage order of the cDNAs. None of the six cDNA clones appear to be candidates for Avr4/6. We conclude that two of these cDNA clones flank a physical region of approximately 24kb and 4.3cM that appears to include the Avr4/6 locus.

Algal Proteins↗
PubMed2002

Inheritance and mapping of 11 avirulence genes in Phytophthora sojae.

Two new crosses involving four races (races 7, 16, 17, and 25) of the soybean root and stem rot pathogen Phytophthora sojae were established (7/16 cross; 17/25 cross). An F2 population derived from each cross was used to determine the genetic basis of avirulence towards 11 different resistance genes in soybean. Avirulence was found to be dominant and determined by a single locus for Avr1b, 1d, 1k, 3b, 4, and 6, as expected for a simple gene-for-gene model. We also observed several cases of segregation, inconsistent with a single dominant gene being solely responsible for avirulence, which suggests that the genetic background of the different crosses can affect avirulence. Avr4 and 6 cosegregated in both the 7/16 and 17/25 crosses and, in the 7/16 cross, Avr1b and 1k were closely linked. Information from segregating RAPD, RFLP, and AFLP markers screened on F2 progeny from the two new crosses and two crosses described previously (a total of 212 F2 individuals, 53 from each cross) were used to construct an integrated genetic linkage map of P. sojae. This revised genetic linkage map consists of 386 markers comprising 35 RFLP, 236 RAPD, and 105 AFLP markers, as well as 10 avirulence genes. The map is composed of 21 major linkage groups and seven minor linkage groups covering a total map distance of 1640.4cM.

Algal Proteins↗