Soil drying as a model for the action of stress factors on the natural bacterial population

The drying of soil samples reduced the abundance (especially of predominant species) and the diversity of bacteria isolated from these samples, making easier the isolation of rare bacterial species. Some bacterial species that were minor before soil drying became dominant in dried soil samples. In general, soil drying allowed the diversity of soil bacteria to be determined more adequately. The bacteria that were isolated from dried soil samples turned out to be resistant to gamma radiation (with LD90 = 2.8-4.6 kGy) and desiccation. It is concluded that soil drying may serve as a model for the action of stress factors on natural bacterial populations. The hypothesis that periodic desiccation was the primary cause of formation of bacterial radioresistance in nature is discussed.     

Soil Drying As a Model for the Action of Stress Factors on Natural Bacterial Populations

The drying of soil samples reduced the abundance (especially of predominant species) and the diversity of bacteria isolated from these samples, making easier the isolation of rare bacterial species. Some bacterial species that were minor before soil drying became dominant in dried soil samples. In general, soil drying allowed the diversity of soil bacteria to be determined more adequately. The bacteria that were isolated from dried soil samples turned out to be resistant to gamma radiation (with LD₉₀ = 2.8–4.6 kGy) and desiccation. It is concluded that soil drying may serve as a model for the action of stress factors on natural bacterial populations. The hypothesis that periodic desiccation was the primary cause of formation of bacterial radioresistance in nature is discussed.     

Competitive Ability and Survival in Soil of Pseudomonas Strain 679-2, a Dominant, Nonobligate Bacterial Predator of Bacteria

A copper-resistant, nonobligate, bacterial predator of bacteria was isolated from soil. It was a Pseudomonas species, designated strain 679-2. It attacked most other nonobligate bacterial predators and hence could control their predatory and other activities in nature. It also inhibited various fungi. It attached to prey cells and produced a toxic, copper-related, growth initiation factor like that produced by Cupriavidus necator. In addition, it produced a second, novel compound that was both antibacterial and antifungal. Strain 679-2 appeared to have only a very limited natural occurrence. It was found only in the soil from one small area in one field. It was absent on the leaves of the plant species that were examined. Regardless of its rarity, however, it was highly competitive in soil. An inoculum consisting of only a few cells added to soil multiplied rapidly to become a major component of the soil microflora within 24 h. A small amount of glutamic acid could be added along with the cells to stimulate production of the toxic compounds noted above, but this was not necessary. After this multiplication, or when large numbers of cells were added to soil, the numbers decreased only slowly during the next several months. Cell survival also was good on plant leaves. The survival in soil and on plant leaves occurred in both laboratory and field experiments. Other than desiccation, the natural mechanism for controlling the numbers or activities of strain 679-2 in soil is not known. The various characteristics of this bacterium, as noted above, are of particular interest because they indicate a possible use of the cells or inhibitor compounds for controlling organisms in soil or on plant surfaces.     

Wide variation in post-emergence desiccation tolerance of seedlings of fynbos proteoid shrubs

Fynbos Proteaceae that are killed by fire and bear their seeds in serotinous cones (proteoids), are entirely dependent on seedling recruitment for persistence. Hence, the regeneration phase represents a vulnerable stage of the plant life cycle. In laboratory-based experiments we investigated the effect of desiccation on the survival of newly emerged seedlings of 23 proteoid species (Leucadendron and Protea) occurring in a wide variety of fynbos habitats. We tested the hypothesis that species of drier habitats would be more tolerant of desiccation than those from more moist areas. Results showed that with no desiccation treatment, or with desiccation prior to radicle emergence, all species germinated to high levels. However, with desiccation treatments imposed after radicle emergence, there were significant declines in seedling emergence after subsequent re-wetting. Furthermore, other than three species that grow in waterlogged habitats, germination responses could not be reliably modeled as a function of soil moisture variables. An important finding was that the species had highly individualistic responses to desiccation. In conclusion, early seedling emergence represents a species-specific stage that is highly sensitive to a decrease in soil moisture. Since species are killed by fire (non-sprouting), vulnerability to increasing aridity associated with anthropomorphic climate change would increase the odds of local and global extinction.     

Isolation and Diversity of Actinomycetes in the Chesapeake Bay

Chesapeake Bay was investigated as a source of actinomycetes to screen for production of novel bioactive compounds. The presence of relatively large populations of actinoplanetes (chemotype II/D actinomycetes) in Chesapeake Bay sediment samples indicates that it is an eminently suitable ecosystem from which to isolate actinomycetes for screening programs. Actinomycetes were isolated from sediment samples collected in Chesapeake Bay with an isolation medium containing nalidixic acid, which proved to be more effective than heat pretreatment of samples. Actinomycete counts ranged from a high of 1.4 × 10⁵ to a low of 1.8 × 10² CFU/ml of sediment. Actinomycetes constituted 0.15 to 8.63% of the culturable microbial community. The majority of isolates from the eight stations studied were actinoplanetes (i.e., chemotype II/D), and 249 of these isolates were obtained in a total of 298 actinomycete isolates. Antimicrobial activity profiles indicated that diverse populations of actinoplanetes were present at each station. DNA hybridization studies showed considerable diversity among isolates between stations, but indicated that actinoplanete strains making up populations at nearby stations were more similar to each other than to populations sampled at distant stations. The diversity of actinoplanetes and the ease with which these organisms were isolated from Chesapeake Bay sediments make this a useful source of these actinomycetes.     

Recovery of photosynthesis in sunflower after a period of low leaf water potential

Photosynthesis was studied in sunflower plants subjected to 1 to 2 days of desiccation and then permitted to recover. The leaf water potential to which leaves returned after rewatering was dependent on the severity of desiccation and the evaporative conditions. Under moderately evaporative conditions, leaf water potential returned to predesiccation levels after 3 to 5 hours when desiccation was slight. Leaf water potentials remained below predesiccation levels for several days after rewatering when leaf water potentials decreased to −13 to −19 bars during desiccation. Leaf water potential showed no sign of recovery when leaf water potentials decreased to −20 bars or below during desiccation. The lack of full recovery of leaf water potential was attributable to increased resistance to water transport in the roots and stem. The resistance ultimately became large enough to result in death of the leaves because net water loss continued even after the soil had been rewatered.     

Drought-deciduous behavior reduces nutrient losses from temperate deciduous trees under severe drought

Nutrient resorption from senescing leaves is an important mechanism of nutrient conservation in temperate deciduous forests. Resorption, however, may be curtailed by climatic events that cause rapid leaf death, such as severe drought, which has been projected to double by the year 2100 in the eastern United States. During a record drought in the southeastern US, we studied 18 common temperate winter-deciduous trees and shrubs to understand how extreme drought affects nutrient resorption of the macronutrients N, P, K, and Ca. Four species exhibited drought-induced leaf senescence and maintained higher leaf water potentials than the remaining 14 species (here called drought-evergreen species). This strategy prevented extensive leaf desiccation during the drought and successfully averted large nutrient losses caused by leaf desiccation. These four drought-deciduous species were also able to resorb N, P, and K from drought-senesced leaves, whereas drought-evergreen species did not resorb any nutrients from leaves lost to desiccation during the drought. For Oxydendrum arboreum, the species most severely affected by the drought, our results indicate that trees lost 50% more N and P due to desiccation than would have been lost from fall senescence alone. For all drought-deciduous species, resorption of N and P in fall-senesced leaves was highly proficient, whereas resorption was incomplete for drought-evergreen species. The lower seasonal nutrient losses of drought-deciduous species may give them a competitive advantage over drought-evergreen species in the years following the drought, thereby impacting species composition in temperate deciduous forests in the future.     

Contents of sugars in leaves of drying desiccation tolerant flowering plants, particularly grasses

Sugar complements were analysed in extracts from leaves of desiccation tolerant species in the angiosperm families Cyperaceae, Gesneriaceae, Liliaceae, Poaceae and Velloziaceae. Total sugar content was higher in live air-dry leaves of all desiccation tolerant species (except the grass Eragrostiella nardoides; 22 µmoles/g dw) than in the dead air-dry leaves of the desiccation sensitive grass Sporobolus pyramidalis (36 µmoles/g dw). Sucrose contents rose to high levels (40–98 µmoles/g dw) in live air-dry leaves of all species (except the grass Eragrostiella nardoides in which it rose to only 11 µmoles/g dw) to become the predominant sugar. Glucose and/or fructose contents frequently were lower after leaf drying but usually these were the sugars of next highest contents in live air-dry leaves. Contents of raffinose (that has been postulated to reduce sucrose crystallization) rose to c. 10% of sucrose contents in air-dry leaves of most desiccation tolerant species (but only c. 4% in Tripogon jacquemontii) compared with c. 2% of sucrose contents in the sensitive grass S. pyramidalis. Trehalose (a rare sugar in seed-plants) was present in all but one desiccation tolerant species (Xerophyta villosa) but only in minor amounts. The results are consistent with the views that sugars play a protective role during drying of desiccation tolerant plants in general but that other factors are also involved indesiccation tolerance, that in desiccation tolerant angiospermae sucrose is generally the predominant protective sugar and that raffinose and trehalose may supplement the role of sucrose.     

Effects of soil fertility on the activities of certain enzymes of desiccated wheat leaves

Summary Isolated wheat (Triticum aestivum var. Kalyan Sona) leaves obtained from plants raised under high and low soil fertility were subjected to quick and slow drying conditions avoiding temperature stress. The leaves of plants raised under high fertility display higher enzyme activity (Amylase, acid pyrophosphatase and ATPase) and large chlorophyll content as compared to the leaves of plants raised under low fertility.It was concluded that adequate nutrition conducive for greater plant vigour brings about an efficient enzyme activity and higher chlorophyll content, despite desiccation, as compared to plants raised under low soil fertility. The results indicated that the hydrature of the tissue need not necessarily be an index of efficiency of metabolism. Under desiccation leaves of plants under high fertility may lose more moisture as compared to leaves of plants raised under low fertility. At the low identical states of hydrature enzyme activities and chlorophyll content remain higher in leaves of plants raised under high soil fertility.     

Distribution of actinomycetes in near-shore tropical marine sediments.

Actinomycetes were isolated from near-shore marine sediments collected at 15 island locations throughout the Bahamas. A total of 289 actinomycete colonies were observed, and all but 6 could be assigned to the suprageneric groups actinoplanetes and streptomycetes. A bimodal distribution in the actinomycete population in relation to depth was recorded, with the maximum numbers occurring in the shallow and deep sampling sites. This distribution can be accounted for by a rapid decrease in streptomycetes and an increase in actinoplanetes with increasing depth and does not conform to the theory that actinomycetes isolated from marine sources are of terrestrial origin. Sixty-three of the isolated actinomycetes were tested for the effects of seawater on growth. Streptomycete growth in nonsaline media was reduced by 39% compared with that in seawater. The actinoplanetes had a near obligate requirement of seawater for growth, and this is presented as evidence that actinomycetes can be physiologically active in the marine environment. Problems encountered with the enumeration of actinomycetes in marine sediments are also discussed.     

Desiccation increases sucrose levels in Ramonda and Haberlea, two genera of resurrection plants in the Gesneriaceae

A few genera of angiosperms are known as 'resurrection plants' since their leaves withstand complete desiccation. In many organisms, including some resurrection plants, desiccation tolerance is associated with the accumulation of special carbohydrates. We examined whether this is also true for the two European angiosperm genera of resurrection plants, Ramonda and Haberlea in the Gesneriaceae. Using gas chromatography, non-structural carbohydrates were determined as a percentage of the dry weight in leaves of Ramonda nathaliae subjected to various desiccation regimes. Sucrose was the predominant soluble carbohydrate in all samples, and its level steadily increased from 2 to 10% during desiccation. Starch amounted to ca 2% in control leaves and disappeared completely within 8 days of desiccation. Considerable amounts (1–2.5%) of raffinose and smaller amounts of its precursor galactinol (1-a-galactosyl- myo -inositol) were present in control leaves; these carbohydrates showed only minor changes upon desiccation. Similar results were obtained when excised leaves of Ramonda nathaliae, Ramonda myconi and Haberlea rhodopensis were subjected to desiccation. These data indicate that sucrose accumulation is connected to desiccation tolerance in Gesneriaceae; the presence of raffinose may be a pre-adaptation since this sugar prevents crystallization of sucrose during drying.     

Distribution of actinomycetes in near-shore tropical marine sediments

Actinomycetes were isolated from near-shore marine sediments collected at 15 island locations throughout the Bahamas. A total of 289 actinomycete colonies were observed, and all but 6 could be assigned to the suprageneric groups actinoplanetes and streptomycetes. A bimodal distribution in the actinomycete population in relation to depth was recorded, with the maximum numbers occurring in the shallow and deep sampling sites. This distribution can be accounted for by a rapid decrease in streptomycetes and an increase in actinoplanetes with increasing depth and does not conform to the theory that actinomycetes isolated from marine sources are of terrestrial origin. Sixty-three of the isolated actinomycetes were tested for the effects of seawater on growth. Streptomycete growth in nonsaline media was reduced by 39% compared with that in seawater. The actinoplanetes had a near obligate requirement of seawater for growth, and this is presented as evidence that actinomycetes can be physiologically active in the marine environment. Problems encountered with the enumeration of actinomycetes in marine sediments are also discussed.     

Hyphae-Colonizing <Emphasis Type="Italic">Burkholderia</Emphasis> sp.—A New Source of Biological Control Agents Against Sheath Blight Disease (<Emphasis Type="Italic">Rhizoctonia solani</Emphasis> AG1-IA<Emphasis Type="Italic">)</Emphasis> in Rice

Sheath blight infection of rice by Rhizoctonia solani Kühn AG1-IA often results in serious yield losses in intensive rice cultivation. Biological control agents (BCAs) have previously been isolated but poor efficiency is often observed when applied under field conditions. This study compares a traditional dual-culture plate assay and a new water-surface microcosm assay for isolation of antagonistic soil bacteria. In the water-surface microcosm assay, floating pathogen mycelium is used as a source for isolation of hyphae-colonizing soil bacteria (HCSB), which are subsequently screened for antagonism. Ten antagonistic soil bacteria (ASB) isolated from a variety of Vietnamese rice soils using dual-culture plates were found to be affiliated with Bacillus based on 16S rRNA gene sequencing. However, all the ASB isolates grew poorly and showed no antagonism in the water-surface microcosm assay. In contrast, 11 (out of 13) HCSB isolates affiliated with Burkholderia sp. all grew well by colonizing the hyphae in the microcosms. Two of the Burkholderia sp. isolates, assigned to B. vietnamiensis based on recA gene sequencing, strongly inhibited fungal growth in both the dual-culture and water-surface microcosm assays; HCSB isolates affiliated to other species or species groups showed limited or no inhibition of R. solani in the microcosms. Our results suggest that HCSB obtained from floating pathogen hyphae can be a new source for isolation of efficient BCAs against R. solani, as the isolation assay mimics the natural habitat for fungal-bacterial interaction in the fields.     

Isolation of new actinomycete strains for the screening of new bioactive compounds

In order to facilitate the discovery of novel bioactive compounds from microorganisms, various techniques for isolation of new actinomycete strains have been attempted. Studies of the vertical distribution of actinomycetes in soil, isolation of actinomycetes from desert soils or fallen leaves, selective isolation of Kitasatospora strains using novobiocin or Actinoplanes strains using the chemotactic method, and the use of gellan gum as a solidifying agent were carried out. We discovered 9 novel bioactive compounds from actinomycete strains isolated under unusual conditions, and proposed two new genera, five new species and one new subspecies.     

STUDIES OF FLUCTUATIONS IN INSECT POPULATIONS XIII. AN IMPROVED METHOD OF ASCERTAINING THE CORRECT DATE TO SAMPLE WHEN ASSESSING LARVAL INFESTATIONS OF THE WHEAT-BLOSSOM MIDGES

Fully fed larvae of the wheat-blossom midges need moist conditions in order to descend from the ears to the soil. A dry spell coinciding with the fully fed stage therefore causes an accumulation of larvae, which subsequently leave the ears in a body when rain comes. Conversely, in wet weather each larva leaves the wheat as soon as it finishes feeding, and, since all the eggs are not laid on the same day, the descent will then be a gradual process.
A method is described of taking small samples of wheat ears at frequent intervals, thus facilitating the determination of the correct date on which to take the main sample for the assessment of infestation by the two wheat-blossom midges.     

Comparative effects of desiccation,heat shock and high temperatures on seed germination of savanna and forest tree species

Although forest and savanna biomes predominate in tropics regions, the factors that control their distribution remain unclear. South American savannas occur in regions that are considered warm and humid enough to support forests, indicating that agents other than climate determine the occurrence of one or the other physiognomy. Herbivory, fire and water deficit have been considered environmental filters that limit the forest species encroachment in savanna physiognomies, but the effects of these filters on the capability of these species to recruit from seeds remain poorly understood. In this study we investigated how stress factors characteristic of savanna environments, such as soil desiccation, heat shocks and high temperatures affect the survival and germination of seeds from savanna and forest tree species. We found that desiccation (to 5%) reduced the germination percentage of forest seeds, but had no effect on the germination of savanna seeds. Forest seeds were less tolerant to heat shocks of 140°C and 200°C, and showed lower germination percentage at temperatures of 35 and 40°C, when compared with savanna seeds. Savanna seeds presented longer germination times and higher germination variance than forest seeds, indicating a risk‐spreading germination strategy among savanna species. The low tolerance of forest seeds to desiccation, heat shock and high temperatures may explain the low recruitment of forest trees into savanna physiognomies. Climate change models predict lower soil moisture, higher temperatures and higher fires frequency for South America biomes. Our results suggest that savanna species are likely to be more capable of withstanding the effects of these changes than forest species.     

Effect of different carbon sources on community composition of bacterial enrichments from soil

Soil is a highly heterogeneous matrix, which can contain thousands of different bacterial species per gram. Only a small component of this diversity (maybe <1%) is commonly captured using standard isolation techniques, although indications are that a larger proportion of the soil community is in fact culturable. Better isolation techniques yielding greater bacterial diversity would be of benefit for understanding the metabolic activity and capability of many soil microorganisms. We studied the response of soil bacterial communities to carbon source enrichment in small matrices by means of terminal restriction fragment length polymorphism (TRFLP) analysis. The community composition of replicate enrichments from soil displayed high variability, likely attributable to soil heterogeneity. An analysis of TRFLP data indicated that enrichment on structurally similar carbon sources selected for similar bacterial communities. The same analysis indicated that communities first enriched on glucose or benzoate and subsequently transferred into medium containing an alternate carbon source retained a distinct community signature induced by the carbon source used in the primary enrichment. Enrichment on leucine presented a selective challenge that was able to override the imprint left by primary enrichment on acetate. In a time series experiment community change was most rapid 18 hours after inoculation, corresponding to exponential growth. Community composition did not stabilize even 4 days after secondary enrichment. Four different soil types were enriched on four different carbon sources. TRFLP analysis indicated that in three out of four cases communities enriched on the same carbon source were more similar regardless of which soil type was used. Conversely, the garden soil samples yielded similar enrichment communities regardless of the enrichment carbon source. Our results indicate that in order to maximize the diversity of bacteria recovered from the environment, multiple enrichments should be performed using a chemically diverse set of carbon sources.     

The role of seed-tuber and stem inoculum in the development of gangrene in potatoes

To determine the relative importance of seed tubers and stems as sources of inoculum for potato gangrene in progeny tubers, different levels of inoculum of Phoma exigua var. foveata were established in field experiments by planting rotting or contaminated seed tubers and by inoculating stems shortly before haulm desiccation. The pathogen was only occasionally detected by isolation from inside green stems in June and July on plants growing from contaminated but unrotted seed. The incidence of pycnidia on desiccated stems increased with increasing concentration of inoculum contaminating seed tubers and with increasing time interval between desiccation and harvest. Stem infection was probably derived from inoculum on seed tubers spreading via the soil to the stem bases. Stem inoculation of plants growing from uninoculated or uncontaminated seed greatly increased both the gangrene potential of progeny tubers (defined as % wounds with gangrene after uniformly damaging tubers and storing them at 5°C for 12 wk), and tuber contamination (defined as % wounds with gangrene after spreading tuber-borne soil onto test tuber slices and storing them at 5°C for 8 wk). However, when stems of plants growing from rotting or contaminated seed were cut at ground level and removed before desiccation, gangrene potential of progeny tubers was only slightly less than that of tubers from untreated plots. There was no evidence that soil inoculum or aerial spread played a significant role in disease development. Gangrene potential and contamination of progeny tubers were related to Contamination levels on seed tubers. Some transmission also occurred from rotted seed tubers to progeny. Inoculum levels around progeny tubers increased rapidly after desiccation even in plots where stems had previously been cut at soil level and removed to eliminate pycnidial development above ground as a source of inoculum. Both gangrene potential and contamination of progeny tubers were greater on cv. Ulster Sceptre than on cv. Pentland Crown. The results showed that the inoculum on seed tubers, whether from rots or surface contamination, contributed more to the contamination of progeny tubers at harvest than did the inoculum from pycnidia on stems following desiccation of the haulm.     

Resistance to desiccation and distribution patterns in bush-dwelling land snails

Bush-dwelling land snails are exposed to desiccating conditions that are more severe than those of snails that seek the shelter of rock crevices, litter or the upper layers of the soil. We studied the resistance to desiccation in four bush-dwelling species of Israeli snails to evaluate a possible relation between their water economy and their distribution pattern. The resistance to desiccation decreased in the following order: Xeropicta vestalis (a widely distributed Mediterranean species), Trochoidea simulata (a desert species), Theba pisana (a Mediterranean species from the sand dunes of the coastal plain), and Monacha haifaensis (a Mediterranean species). Xeropicta vestalis also had the quickest response to the desiccating conditions. It is probably prevented from establishing itself in the desert, in spite of its superior water economy, because it is an annual, semelparous species, and the desert is a highly unpredictable environment. An immediate response to desiccating conditions may be a key factor in the success of desert-inhabiting land snails. Snails from more humid regions take several days to recruit their water preserving mechanisms, a delay which may be crucial to their water balance. The conchiometrics of X. vestalis and T. simulata suggest that the main water preserving mechanisms of these species are located in the mantle, rather than in the shell and epiphragm.