Strains Of Xanthomonas Campestris Pv Research Articles

Lipopolysaccharides and plant responses to phytopathogenic bacteria

Abstract Treatment of the leaves of pepper (Capsicum annuum) cv. ECW10R with lipopolysaccharides (LPS) from both plant pathogenic and enteric bacteria alters several aspects of the plant response to subsequent inoculation with phytopathogenic xanthomonads. LPS pre-treatment prevents the hypersensitive reaction caused by strains of Xanthomonas campestris pv. vesicatoria carrying the avirulence gene avrBs1 (a gene-for-gene interaction) and by X. campestris pv. campestris (a non-host interaction). Associated with this effect are the earlier synthesis of feruloyl- and coumaroyl-tyramine, phenolic conjugates that are potentially antimicrobial, and alterations in the expression patterns of genes for some pathogenesis-related (PR) proteins. Similar effects on the timing of phenolic conjugate synthesis are also seen in the compatible interaction with X. campestris pv. vesicatoria, although the level of the response is lower. Recognition of LPS by plants may allow expression of resistance in the absence of catastrophic tissue damage. However phytopathogenic bacteria may have evolved mechanisms to suppress the effects of LPS (and of other non-specific bacterial elicitors) on plant cells.

Effect of Temperature on the Incubation Period and Leaf Colonization in Bacterial Blight of Anthurium

ABSTRACT Effect of temperature on leaf colonization in anthurium blight was studied using a bioluminescent strain of Xanthomonas campestris pv. dieffenbachiae. In a susceptible cultivar, colonization of leaf tissues (monitored by detection of bioluminescence) and symptom development (assessed visually) advanced rapidly at higher temperatures. For a susceptible cultivar, there was a linear relationship between degree-days and percent leaf area colonized by the pathogen, indicating that leaf colonization in a susceptible cultivar was a direct function of the cumulative effect of temperature. The degree-day intercept of the regression line represented the time from inoculation to detection of bioluminescence, and the slope indicated the increase of leaf colonization per degree-day. There also was a linear relationship between the logarithm of degree-days and the logarithm of percent leaf area showing visible symptoms in a susceptible cultivar. The degree-day intercept of this relationship represented the incubation period (about 500 degree-days). The degree-days required to detect bioluminescence was not considerably different between susceptible and resistant cultivars. However, the subsequent rates of leaf colonization were significantly lower for a resistant cultivar than for a susceptible cultivar in all temperature regimes. The results suggest that multiplication of the pathogen in the leaf tissues is optimized in the susceptible cultivar. In contrast, in the resistant cultivar, the defense mechanisms overshadow the temperature effect. The differential response to temperatures may be an additional indicator of cultivar susceptibility.

Specific detection of Xylella fastidiosa Pierce's disease strains.

Pierce's disease (PD, Xylella fastidiosa) of grapevine is the primary pathogen limiting vinifera grape production in Florida and other regions of the southeastern United States. Quick and accurate detection of PD strains is essential for PD studies and control. A unique random amplified polymorphic DNA (PD1-1-2) was isolated from a PD strain from Florida. Fragment PD1-1-2 was cloned, sequenced, and found to be 1005 bp in length. PCR primers were designed to utilize these sequence data for PD strain detection. One primer set (XF176f-XF954r) amplified a 779-bp DNA fragment from 34 PD strains including seven pathotypes of X. fastidiosa, but not from strains of Xanthomonas campestris pv. campestris, Xan. vesicatoria or Escherichia coli. A second primer set (XF176f and XF686r) amplified a 511-bp fragment specific to 98 PD strains, but not from strains of citrus variegated chlorosis, mulberry leaf scorch, oak leaf scorch, periwinkle wilt, phony peach, or plum leaf scald. Sequence analysis indicated that RAPD fragment PD1-1-2 contains a Ser-tRNA gene. The PD-specific region includes a TaqI restriction site (TCGA) and is 150 bp downstream of the Ser-tRNA gene.

First Observation of Xanthomonas campestris pv. vesicatoria Race T2P7 Isolated from Pepper in the Philippines

Thirteen strains of Xanthomonas campestris pv. vesicatoria, causal agent of bacterial spot of pepper (Capsicum spp.) and tomato (Lycopersicum spp.), were isolated from six different pepper fields in the vicinity of Bongabon, Luzon, the Philippines. The strains were characterized based on sensitivity to copper and streptomycin, physiological tests, analysis of whole cell fatty acids (fatty acid methyl ester; FAME), serology, amplification of the 16S-23S rDNA spacer region, pathogenicity, and virulence on pepper and tomato differential genotypes as previously described (1,2). All of the Philippine strains were copper resistant (50 μg/ml), and streptomycin sensitive (20 μg /ml). None of the strains were pectolytic or amylolytic. FAME analysis showed that all strains had similar fatty acid profiles, and matched the strains to X. campestris (similarity indices [S.I.] = 0.186-0.455). With the universal primers 4F/7R, a DNA fragment (680 bp) was amplified from 16S-23S rDNA spacer regions of all Philippine strains. Serological fingerprints of the strains with a set of eight monoclonal antibodies (MAbs) in indirect enzyme-linked immunosorbent assay showed that all Philippine strains were serovar A1. Existence of the avrRxv gene in X. campestris pv. vesicatoria race T1 strains was also confirmed for all Philippine strains by amplifying a DNA fragment (approximately 0.7 kb) with the primer set RST 27/28. This is genetic evidence for race identity of the Philippine strains with T1. However, hypersensitive test results on pepper and tomato differential lines identified the Philippine strains as T2P7. These results showed that X. campestris pv. vesicatoria strains may contain the avrRxv gene, but it may not be functional. This is the first report of the observation of X. campestris pv. vesicatoria race T2P7 on pepper.

Phenotypic characteristics of Xanthomonas campestris pv. campestris from Nepal

Twenty strains of Xanthomonas campestris pv. campestris (Xcc) were isolated from two major crucifer-growing valleys, Chitwan and Kathmandu in Nepal and characterized by biochemical and pathogenicity tests. Strains were homogeneous in bacteriological characteristics. The ability of a strain to induce high or low disease severity index (DSI) on three host plants, broccoli, cabbage, and cauliflower, was interpreted as virulence. Strains that were associated with high or low virulence were significantly different (P>0.05). No relationship between virulence and biochemical characteristics was observed.

Bacterial Spot of Tomato and Pepper on Four East Caribbean Islands: Races, Their Abundance, Distribution, Aggressiveness, and Prospects for Control.

A total of 5,134 strains of Xanthomonas campestris pv. vesicatoria obtained from diseased pepper and tomato fruits at 22 diverse locations on the East Caribbean islands of Antigua, St. Christopher, St. Lucia, and Grenada were assessed for pathogenicity, taxonomic group phenotypes, and sensitivity to copper, zinc, and streptomycin. Altogether, 15 known races, namely T1, T2, P0T1, P0T2, P1T1, P1T2, P2T1, P2T2, P3T1, P3T2, P4T2, P5T2, P5T3, P6T2, and P6T3, were detected. Races T1, P0T1, P0T2, P1T2, P2T1, and P3T1 were common to all islands except St. Lucia. St. Lucia had races P2T2, P3T2, P4T2, and P6T2 in common with Grenada and two races each from among the four pathotypes in common with Antigua and St. Lucia. Races T1, P1T2, P3T1, and P3T2 were the most abundant strains of the bacterium in Antigua, whereas the two latter pathotypes, as well as races P0T1 and P2T1, were among the most prevalent strains in St. Christopher. In contrast, races P6T2 and P5T2 predominated in St. Lucia, and P6T2, P1T2, and P3T2 were the abundant pathotypes in Grenada. Generally, the largest bacterial spot fruit lesions were associated with the predominant races on each island, and the smallest lesions were generally linked to races capable of overcoming gene Bs2. Only these latter races were similar to group B strains of X. campestris pv. vesicatoria with respect to the presence of β-protein in their cellular protein profiles, ability to hydrolyze starch, and inability to oxidize cis-aconitate. Other race types tested resembled group A strains of the bacterium in their ability to utilize cis-aconitate, inability to hydrolyze starch and pectate, and the presence of the α-protein in profiles of cellular proteins. Copper-resistant strains of X. campestris pv. vesi-catoria constitute 8.7 to 10.6% of the pathogen collections from Antigua and St. Christopher and 44.4 and 66.7% of pathotypes from St. Lucia and Grenada, respectively. Almost all strains were sensitive to zinc. Streptomycin resistance was observed among 62% of the strains in St. Christopher and Grenada and 62.8 to 83.6% of those in Antigua and St. Lucia.

Molecular characterization of the incitant of cowpea bacterial blight and pustule, Xanthomonas campestris pv. vignicola

Strains of Xanthomonas campestris pv. vignicola (Xcv), isolated from cowpea leaves with blight or minute pustules and collected from various geographic areas, were selected on the basis of pathological and physiological features. All strains were analyzed for genotypic markers by two methods: ribotyping with EcoRI endonuclease, and RFLP analysis with a plasmid probe (pthB) containing a gene required for pathogenicity from Xanthomonas campestris pv. manihotis. Ribotyping revealed a unique pattern for all the strains that corresponded to the previously described ribotype rRNA7. Based on polymorphism detected by pthB among Xcv strains, nine haplotypes were defined. The observed genetic variation was independent of the geographic origin of the strains and of pathogenic variation. Some haplotypes were widely distributed, whereas others were localized. In some cases, we could differentiate strains isolated from blight symptoms and pustules according to haplotypic composition. However, in most cases, no significant differences were observed. Our results and the previous pathogenic and biochemical characterizations suggest that the strains isolated from leaves with blight symptoms or minute pustules belong to the same pathovar. We provide information on pathogen diversity that can be used to identify and characterize resistant germplasm.

Cloning and Characterization of a Member of theXanthomonas avr/pthGene Family That Evades All Commercially Utilized CottonRGenes in the United States

ABSTRACT The highly virulent African strains of Xanthomonas campestris pv. malvacearum are quarantined pathogens in the United States and can evade or overcome all commercially utilized resistance (R) genes in cotton grown in the United States including the entire set of host differential lines used to distinguish 19 races of the pathogen. Nevertheless, the African strains carry multiple DNA fragments that strongly hybridize with members of the Xanthomonas avirulence (avr)/pathogenicity (pth) gene family. Since all previously tested members of the gene family confer avirulence against one or more R genes in cotton, strains carrying multiple members might not be expected to evade so many different R genes. The hybridizing DNA fragments were cloned from African strain XcmN and found to confer water-soaking ability to a nearly asymptomatic mutant strain of the pathogen. Restriction mapping, Southern hybridization, and DNA sequencing of the cloned fragments from XcmN were used to identify two water-soaking genes, pthN and pthN2, as new members of the Xanthomonas avr/pth gene family. The complete DNA sequence of pthN was obtained, and it is >94% identical with all other sequenced members of the gene family. Gene fusions of pthN with avrb6 (another family member) and other experiments revealed that the ability of African strain XcmN to water-soak cotton and avoid recognition by commercially used cotton R genes is determined by the specific repeats of multiple functional members of the Xanthomonas avr/pth gene family.

Cytochemistry of defense responses in cassava infected by Xanthomonas campestris pv. manihotis

Stems of susceptible and resistant cassava plants have been cytologically investigated for their defense reactions to an aggressive strain of Xanthomonas campestris pv. manihotis. Histochemistry, in conjunction with gold cytochemistry, revealed that in susceptible and resistant plants, phloem and xylem parenchyma cells displayed a wide range of responses that limited the bacterial growth within the infected plants. Lignification and suberization associated with callose deposition were effective mechanisms that reinforced host barriers in the phloem. In the infected xylem, vessels were plugged by a material of pectic and (or) lignin-like origin. Flavonoids have been seen to be incorporated in secondary cell wall coatings. These reactions occurred at a higher intensity in the resistant plants. The number of phoem and xylem cells producing autofluorescent compounds was higher in infected resistant plants than in susceptible plants. Reactions have been observed in the resistant variety only, such as secretion of phenol-like molecules by tyloses and hyperplasic activity of phloem cells that compartmentalized bacterial lysis pockets, which are potent secondary inoculum sources.Key words: lignin, suberin, callose, phenol, tylose, flavonoid, pectin.

USE OF BIOLUMINESCENCE FOR THE DETECTION OF THE EFFECTIVENESS OF BIOLOGICAL CONTROL AGENTS ON BLACK ROT DISEASE OF CABBAGE

A genetically engineered, bioluminescent strain of Xanthomonas campestris pv. campestris (Xcc) was used to study the effectiveness of plant growth-promoting rhizobacteria (PGPR) as disease control agents. Black-rot-susceptible cabbage plants were wound-inoculated with PGPR and wound- or mist-inoculated with bioluminescent Xcc 10 days later. Growth of the bioluminescent strain in the plants was followed over time with a low-light, charge-coupled device camera. Several PGPR strains effectively reduced growth of the bioluminescent pathogen in the plants when bacteria were introduced into the plant by wound. PGPR inoculation was less effective when bioluminescent bacteria were introduced into the plant by mist inoculation. Little effect on symptom reduction was observed.

Isoenzyme diversity in Xanthomonas campestris pv. mangiferaeindicae

A collection of 26 strains of Xanthomonas campestris pv. mangiferaeindicae isolated from three different host species in eight countries was investigated for variation in isozyme patterns. Three enzyme systems were analysed: esterase (EST), phosphoglucomutase (PGM) and superoxide dismutase (SOD). Four groups of strains were identified: nonpigmented strains isolated from mango and pepper‐tree in Australia, Comores, India, Reunion Island, South Africa, and Taiwan; nonpigmented Brazilian strains from mango; nonpigmented strains from ambarella isolated in the French West Indies; heterogeneous yellow pigmented strains from mango (Brazil and Reunion Island). The value of isozyme profiling as markers of the pathogenicity groups in X. c. pv. mangiferaeindicae is discussed.

Relationship between Symptom Development and Actual Sites of Infection in Leaves of Anthurium Inoculated with a Bioluminescent Strain of Xanthomonas campestris pv. dieffenbachiae

The infection process of bacterial blight of anthurium was monitored with a bioluminescent strain of Xanthomonas campestris pv. dieffenbachiae. The relationship between symptom expression on infected leaves (assessed visually) and the extent of bacterial movement within tissues (evaluated by bioluminescence emission) varied among anthurium cultivars. In several cultivars previously considered susceptible on the basis of symptom development alone, bacterial invasion of leaves extended far beyond the visually affected areas. In other cultivars previously considered resistant, bacterial invasion was restricted to areas with visible symptoms. In three cultivars previously considered resistant, leaves were extensively invaded by the bacterium, and yet few or no symptoms were seen on infected leaves. The pathogen was consistently recovered from leaf sections emitting bioluminescence but not from sections emitting no light. At an early stage of infection, no significant differences in the percentages of infected areas as determined by visual assessment were observed in any of the cultivars. However, differences among cultivars were detected by bioluminescence as the disease progressed, because bacterial invasion was not always accompanied by symptom expression. In susceptible cultivars, the advancing border of infection was 5 to 10 cm inward from the margins of the visible symptoms and often reached to the leaf petiole even when symptoms were visible in <10% of the total leaf area. Comparisons of anthurium cultivars in which a nondestructive method was used to quantify the severity of leaf infection by a bioluminescent pathogen have enabled us to evaluate susceptibility and resistance to bacterial blight accurately. Such evaluations will be of importance in breeding resistant cultivars for disease control.

Role of extracellular polysaccharides from Xanthomonas campestrispv. vesicatoriain bacterial spot of pepper

Purified extracellular polysaccharides (EPS) yielding mainly glucose and mannose on hydrolysis, were prepared from culture filtrates of three strains of Xanthomonas campestrispv. vesicatoriaand tested at concentrations of 2.5 and 5.0mg ml -1in water on four pepper cultivars, either alone or in combination with the bacterium. Pathogen growth and host responses including watersoaking, electrolyte leakage, discoloration and collapse of leaves were studied. EPS alone induced watersoaking and in some cases foliar discoloration including yellowing, browning and /or necrosis. Highly viscous EPS produced by one strain of the bacterium and characterized by a viscosity of 4.13 ×10 -4N-SM 2at 5mg ml -1generally delayed host response to infection and pathogen growth. The other EPS of viscosities ranging from 5.24 ×10 -5to 2.06 ×10 -4N-SM 2at a similar concentration accelerated leaf collapse, maximum electrolyte leakage and visible symptoms of bacterial spot during the incompatible and compatible response. These EPS also inhibited or accelerated pathogen growth during incompatible and compatible responses respectively.

Survival and persistence of bioluminescent Xanthomonas campestris pv. campestris on host and non‐host plants in the field environment

Dispersal and persistence of a pathogenic strain of Xanthomonas campestris pv. campestris, genetically engineered to bioluminesce, was followed in and on host and non‐host plants in the field environment. Black rot susceptible cabbage plants were mist inoculated with the bioluminescent strain only, or were mist inoculated with X. campestris pv. vesicatoria or a weakly pathogenic strain of X. c. campestris 1 week before challenge inoculation with the bioluminescent strain. Growth of the bioluminescent strain was detected with a low‐light, charge‐coupled device camera or through bioluminescence measurements of broth‐enrichment cultures of leaf disk samples. Bioluminescent X. c. campestris could often be observed as populations on symptomless leaves or in lesions, and persisted as a vascular endophyte for more than 6 months throughout the winter growing season. Dispersal to cruciferous and non‐cruciferous weeds was frequently detected. Pre‐inoculation with X. c. vesicatoria or the weakly pathogenic X. c. campestris did not significantly affect the movement and persistence of the bioluminescent strain nor reduce the incidence of black rot disease.

Studies on a Quantitative Analysis of Oryzalides and Oryzalic Acids in Rice Plants by GC-SIM

A procedure for the quantitative analysis of oryzalides and oryzalic acids in leaves of rice cultivars was established by using GC-MS-selective ion monitoring (GC-SIM), 17-deuterated oryzalide B being used as an internal reference. The total contents of four compounds of oryzalides and oryzalic acids in the leaves at three different growth stages of Norin-27, i.e., seedling, tillering, and ripening, showed values of 5.3, 17.9, and 37.9 μg/g, respectively. The contents of oryzalides and oryzalic acids in each organ, i.e., leaf, stem, ear, and root, showed values of 35.0 and 1.7 μg/g for the former two, respectively, and were not detectable for the latter two. A preliminary analysis of Norin-27 and Rantai emas leaves inoculated with an incompatible and compatible strain of Xanthomonas campestris pv. oryzae showed a greater accumulation of oryzalides and oryzalic acids, especially in the lesion area, than the figures for healthy leaves. Mechanical injury by a needle-bundle also increased the total amounts.

Copper accumulation in Xanthomonas campestris pv. vesicatoria

A strain of Xanthomonas campestris pv. vesicatoria showing resistance to 1.2 mM cupric sulfate was analyzed by atomic absorption spectroscopy and ESI (electron spectrophotometry imaging). Accumulation of copper was detected in the periphery of the cell membrane region, suggesting that the mechanism of copper resistance is similar to that previously described for Pseudomonas species. The ESI technique was used to detect copper in the membrane region. Copper-resistance in X. campestris pv. vesicatoria 484 is inducible and occurs by accumulation of the metal and not by efflux mechanism as has been suggested. The growth curve also showed that this system is inducible.

A 13C-Pulse-labeling study of phytoalexin biosynthesis in hypersensitively responding cotton cotyledons

Time courses of multiplication of an incompatible strain of Xanthomonas campestris pv. malvacearum, accumulation of phytoalexins, and development of phytoalexin-rich, yellow-green fluorescent, hypersensitively necrotic cells were followed in cotton cotyledons. Occasionally, hypersensitively responding palisade cells were seen to pass through a stage in which the yellow-green fluorescence, due to the phytoalexins lacinilene C and its methyl ether, was confined to the cytoplasmic layer surrounding an intact vacuole, but more often, when cells became yellow-green fluorescent, the fluorescence appeared throughout the cell. The period of most rapid increase in fluorescent cell numbers, 48-72 h post-inoculation, was also the period of most rapid increase in phytoalexin content, and the rate of bacterial multiplication began to slow down at this time. Stable 13C isotope incorporation from a 4 h pulse with [13C2]acetate into the phytoalexin 2,7-dihydroxycadalene and its methyl ether was measured by direct probe electron impact-mass spectrometry to monitor their biosynthesis at 2, 3, 4, and 6 days after inoculation. Highest incorporation rates occurred during the period of most rapid increase in numbers of fluorescent cells, consistent with the hypothesis that these cells achieve a burst of phytoalexin biosynthesis causing them to become fluorescent as they die. In addition, biosynthesis was still active when fluorescent cell numbers approached their maximum and was appreciably above zero 1 and 2 days later, which suggests the possibility that healthy cells neighboring the hypersensitively responding ones contribute to the biosynthesis of phytoalexins in cotton.

Identification of Xanthomonas campestris pv. Juglandis from (Persian) English Walnut Nursery Trees in South Africa

AbstractBacterial isolates producing yellowish colonies on Nutrient Agar were recovered from symptoms of suspect walnut blight disease on leaves of nursery trees in the southwestern Cape Province of South Africa. The isolates were identified by pathogenicity tests on leaves of walnut and plum trees in the greenhouse. Fifteen isolates from four cultivars at two nurseries produced typical lesions of blight on walnut and one isolate. typical lesions of bacterial spot disease on plum leaves. Cluster analysis was done on 28 characteristics recorded from colony growth. colour. form. and elevation on four different culture media, and starch hydrolysis on a semi‐selective medium for the isolation of Xanthomonas campestris pv. juglandis. Total DNA of the isolates was digested with restriction endonuclease Spel and resolved by contour‐clamped homogeneous electric field (CHEF) electrophoresis. Two phenotypic clusters were distinguished among the 15 South African and one reference strain of X.c.pv. juglandis at the 54%Ssm level. The isolate which induced disease symptoms on plum grouped with reference strains of Xanthomonas campestris pv. pruni in a third cluster. Two‐thirds of the isolates were not characterized on the semi‐selective medium for X.c. pv. juglandis. DNA restriction fragment banding patterns were similar for most isolates of X.c.juglandis in the same phenotypic cluster. However, DNA banding patterns were non‐distinct for some isolates with similar phenotypic characters. Phenotypic characteristics and DNA restriction fragment banding patterns of the isolates were not correlated with geographical origin or cultivar specificity.

Difference Between Xanthomonas campestris pv. phlei and X.c pv. graminis Revealed by Genomic Fingerprinting and Restriction Fragment Length Polymorphism Patterns

AbstractGenomic DNA was prepared from 16 strains of Xanthomonas campestris pv. graminis and Xanthomonas campestris pv. phlei isolated from six species of forage grasses in four countries. The two pathovars could be distinguished clearly by genomic fingerprints generated by EcoRI, BamHI or HindIII digestion. DNA profiles produced by HindIII digestion could differentiate not only between the two pathoars but also among strains within the same pahtovar from different countries. A 1.6 kb EcoRI fragment was cloned from genomic DNA of strain LMG726 and used to detect restriction fragment‐length polymorphism among the same strains. EcoRI and BamHI polymorphisms were seen between the two pathovars probed with this 1.6 kb EcoRI fragment (p726EI probe). These polymorphisms appeared to be highly conserved and unique for each pathovar, consistent with previous grouping of the strains based on other criteria.

A Third Tomato Race ofXanthomonas campestrispv.vesicatoria

Strains of Xanthomonas campestris pv. vesicatoria were isolated from several tomato fields in Florida that differed from tomato race 1 (T1) strains commonly found in Florida. The strains produced a compatible reaction on tomato genotype Hawaii 7998 (H7998), were amylolytic and pectolytic, and were classified originally as T2 strains. However, these strains produced a rapid hypersensitive response when infiltrated into the tomato genotypes Hawaii 7981 (H7981), PI 126932, and PI 128216, whereas T2 strains produced a compatible reaction. In an experiment where electrolyte leakage was determined in leaflets of tomato cv. Bonny Best (compatible with all tomato strains), H7998 (resistant to T1 strains), and the two PIs, a member of these new strains induced a rapid hypersensitive response in the three tomato genotypes, whereas a T2 strain did not. After low concentrations of bacteria were infiltrated into the mesophyll of leaflets of PI 126932, PI 128216, H7998, and cv. Walter (compatible with all tomato strains), internal populations of the amylolytic, pectolytic Florida strain were reduced when tested in the first two genotypes but not in the latter two genotypes. Populations of the T1 strain were reduced only in H7998. These new strains are designated tomato race 3 (T3). When the T3 strains were compared with a representative group of T1 and T2 strains by fatty acid analysis and carbon substrate utilization patterns, the new T3 strains clustered distinctly from representative T1 and T2 strains. The evidence suggests the T3 strain is a recent introduction to Florida. An amylolytic, pectolytic strain isolated from seed grown in Thailand reacted similarly to the T3 strains on the tomato differentials. It also clustered with the T3 strains in the fatty acid and carbon substrate utilization dendrograms