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1.  Chrysanthemum Cutting Productivity and Rooting Ability Are Improved by Grafting 
The Scientific World Journal  2013;2013:286328.
Chrysanthemum has been commercially propagated by rooting of cuttings, whereas the quality will decline over multiple collections from a single plant. Therefore, we compared the vigour, rooting ability, and some physiological parameters between cuttings harvested from nongrafted “Jinba” (non-grafted cuttings) with those collected from grafted “Jinba” plants onto Artemisia scoparia as a rootstock (grafted cuttings). The yield, length, node number, stem diameter, fresh weight, and dry weight of the grafted cuttings were superior to the non-grafted cuttings. Also grafted cuttings “Jinba” rooted 1 day earlier, but showing enhanced rooting quality including number, length, diameter, and dry weight of roots, where compared to the non-grafted. The physiological parameters that indicated contents of soluble protein, peroxidase activity, soluble sugar, and starch, ratios of soluble sugar/nitrogen ratio, and carbohydrate/nitrogen (C/N), as well as contents of indole-3-acetic acid (IAA) and abscisic acid (ABA), and IAA/ABA ratio were significantly increased in the grafted cuttings. This suggested their important parts in mediating rooting ability. Results from this study showed that grafting improved productivity and rooting ability related to an altered physiology, which provide a means to meet the increasing demand.
PMCID: PMC3708427  PMID: 23878523
2.  Crystal structure of stable protein CutA1 from psychrotrophic bacterium Shewanella sp. SIB1 
Journal of Synchrotron Radiation  2010;18(Pt 1):6-10.
The crystal structure of CutA1 from the psychrotrophic bacterium Shewanella sp. SIB1 in a trimeric form was determined at 2.7 Å resolution. This is the first crystal structure of a psychrotrophic CutA1.
CutA1 is widely found in bacteria, plants and animals, including humans. The functions of CutA1, however, have not been well clarified. It is known that CutA1s from Pyrococcus horikoshii, Thermus thermophilus and Oryza sativa unfold at temperatures remarkably higher than the growth temperatures of the host organisms. In this work the crystal structure of CutA1 from the psychrotrophic bacterium Shewanella sp. SIB1 (SIB1–CutA1) in a trimeric form was determined at 2.7 Å resolution. This is the first crystal structure of a psychrotrophic CutA1. The overall structure of SIB1–CutA1 is similar to those of CutA1 from Homo sapiens, Escherichia coli, Pyrococcus horikoshii, Thermus thermophilus, Termotoga maritima, Oryza sativa and Rattus norvergicus. A peculiarity is observed in the β2 strand. The β2 strand is divided into two short β strands, β2a and β2b, in SIB1–CutA1. A thermal denaturation experiment revealed that SIB1–CutA1 does not unfold completely at 363 K at pH 7.0, although Shewanella sp. SIB1 cannot grow at temperatures exceeding 303 K. These results indicate that the trimeric structural motif of CutA1 is the critical factor in its unusually high stability and suggest that CutA1 needs to maintain its high stability in order to function, even in psychrotrophs.
PMCID: PMC3004244  PMID: 21169681
CutA1; Shewanella sp. SIB1; crystal structure; thermal denaturation; trimeric structural motif
3.  Uptake of bacteriophage f2 through plant roots. 
A model system was designed to measure viral uptake through the roots of plants and translocation to distal plant parts. For this study, uptake of bacteriophage f2 was measured in corn and bean plants growing in hydroponic solutions. Few phage were detected in plants with uncut roots. However, when roots of both plant types were cut just before exposure to very high concentrations of phage, the amount of phage uptake was several orders of magnitude greater than with uncut roots, but still was considerably less than that which was theoretically possible. Furthermore, cut roots were rapidly repaired, thus inhibiting uptake, and the amount of uptake in plants with cut roots was proportional to phage exposure levels. Finally, phage were transported to all plant parts examined, but their survival times within each portion of the plants appeared to be of limited duration. All of these factors tend to minimize the possible public health significance associated with viral uptake through the root systems of plants.
PMCID: PMC244192  PMID: 7103476
4.  Effect of Cutting Age on the Resistance of Prunus cerasifera (Myrobalan Plum) to Meloidogyne arenaria 
Journal of Nematology  1995;27(4S):634-638.
The response of softwood cuttings of Myrobalan plum infested after 50 and 105 days with 3,000 second-stage juveniles (J2) of Meloidogyne arenaria was compared to 15-month-old hardwood cuttings in 13 genotypes ranging from highly resistant to susceptible. Gall index and number of galls were recorded 30 days after infestation. Fifty-day-old cuttings rooted in perlite developed many rootlets, but had only incipient galls after infestation. In sand, rooting of 50-day-old cuttings not treated with indolebutyric acid (IBA) hormone was so variable that their resistance could not be assessed. Similar cuttings rooted with IBA developed more galls, but neither number of galls per plant nor gall index was a reliable criterion for determination of host suitability. Because of the better rooting results with IBA treatment, 105-day-old cuttings were first rooted with IBA in perlite and then transferred into sand for nematode inoculation. Known highly resistant genotypes of Myrobalan plum were gall-free and the responses of other genotypes paralleled that of the reference hardwood cuttings, although the test was less discriminating. Expression of M. arenaria host suitability in Myrobalan plum depends on root tissue maturation and cannot be reliably evaluated with 50-day-old cuttings.
PMCID: PMC2619659  PMID: 19277333
hardwood cutting; host suitability; Meloidogyne arenaria; nematode; Prunus cerasifera; resistance; root galling; root-knot nematode; softwood cutting
5.  Effect of Mowing Cotton Stalks and Preventing Plant Re-Growth on Post-Harvest Reproduction of Meloidogyne incognita 
Journal of Nematology  2010;42(2):96-100.
The southern root-knot nematode (Meloidogyne incognita) is a major parasite of cotton in the U.S., and management tactics for this nematode attempt to minimize population levels. We compared three post-harvest practices for their ability to reduce nematode population levels in the field, thereby reducing initial nematode population for the next year's crop. The three practices tested were: 1) chemical defoliation before harvest plus cutting cotton stalks after harvest, 2) chemical defoliation plus applying a herbicide to kill plants prior to cutting the stalks, and 3) chemical defoliation without cutting stalks. Experiments were conducted in both the greenhouse and in the field. The greenhouse experiments demonstrated that M. incognita reproduction (measured as egg counts and root gall rating indices) was significantly greater when stalks were not cut. Cutting stalks plus applying herbicide to kill cotton roots did not significantly reduce nematode reproduction compared to cutting stalks alone. In field experiments, cutting stalks reduced egg populations and root galling compared to defoliation without stalk cutting. In a greenhouse bioassay which used soil from the field plots, plants grown in soil from the defoliation only treatment had greater root gall ratings and egg counts than in the stalk cutting plus herbicide treatment. Therefore, we conclude that cutting cotton stalks immediately after harvest effectively reduces M. incognita reproduction, and may lead to a lower initial population density of this nematode in the following year.
PMCID: PMC3380473  PMID: 22736845
Cotton; cultural control; defoliation; Gossypium hirsutum; herbicide; Meloidogyne incognita; nematode management; post-harvest; reproduction; roots; southern root-knot nematode
6.  Discriminative feeding behaviour of Anopheles gambiae s.s. on endemic plants in western Kenya 
Medical and veterinary entomology  2007;21(1):103-111.
Anopheles gambiae Giles s.s. (Diptera: Culicidae) is known to feed on plant sugars, but this is the first experimental study to consider whether it discriminates between plant species. Thirteen perennial plant species were selected on the basis of their local availability within the vicinity of human dwellings and larval habitats of An. gambiae s.s. in western Kenya. Groups of 100 or 200 mosquitoes were released into cages either with a cutting of one plant type at a time (single-plant assay) or with cuttings of all 13 plants simultaneously (choice assay), respectively, and left overnight. In the choice assay, direct observations of the percentages of mosquitoes perching or feeding on each plant were recorded over four 1-h periods each night. For both types of assay, mosquitoes were recaptured and the percentage that had fed on plants was assessed by testing them individually for the presence of fructose. To identify which plants the choice-assay mosquitoes had fed on, gas chromatography (GC) profiles of samples of mosquito homogenates were compared with GC profiles of extracts from relevant parts of each plant. Four of the plants that were observed to have been fed on most frequently in the choice assay (Parthenium hysterophorus L., Tecoma stans L., Ricinus communis L., and Senna didymobotrya Fresen) were also shown to have been ingested most often by mosquitoes in both types of assay, suggesting that An. gambiae is differentially responsive to this range of plants, regardless of whether the plants were presented singly or mixed together. Significantly more females than males fed on plants, with the exception of P. hysterophorus L., one of the plants most frequently fed on. For most plant species (ten of 13), GC profiles indicated that An. gambiae obtained sugars primarily from flowers. The exceptions were P. hysterophorus L., Lantana camara L. and R. communis L., on which An. gambiae fed more often from leaves and stems than from flowers.
PMCID: PMC2705332  PMID: 17373953
Anopheles gambiae; Kenya; malaria; nectar; plant-feeding
7.  Assessment of Variation in Bacterial Composition among Microhabitats in a Mangrove Environment Using DGGE Fingerprints and Barcoded Pyrosequencing 
PLoS ONE  2012;7(1):e29380.
Here, we use DGGE fingerprinting and barcoded pyrosequencing data, at six cut-off levels (85–100%), of all bacteria, Alphaproteobacteria and Betaproteobacteria to assess composition in the rhizosphere of nursery plants and nursery-raised transplants, native plants and bulk sediment in a mangrove habitat. When comparing compositional data based on DGGE fingerprinting and barcoded pyrosequencing at different cut-off levels, all revealed highly significant differences in composition among microhabitats. Procrustes superimposition revealed that ordination results using cut-off levels from 85–100% and DGGE fingerprint data were highly congruent with the standard 97% cut-off level. The various approaches revealed a primary gradient in composition from nursery to mangrove samples. The affinity between the nursery and transplants was greatest when using Betaproteobacteria followed by Alphaproteobacteria data. There was a distinct secondary gradient in composition from transplants to bulk sediment with native plants intermediate, which was most prevalent using all bacteria at intermediate cut-off levels (92–97%). Our results show that PCR-DGGE provides a robust and cost effective exploratory approach and is effective in distinguishing among a priori defined groups.
PMCID: PMC3256149  PMID: 22247774
8.  Identification of Xylem Occlusions Occurring in Cut Clematis (Clematis L., fam. Ranunculaceae Juss.) Stems during Their Vase Life 
The Scientific World Journal  2012;2012:749281.
During the vase life of cut stems obstruction of xylem vessels occurs due to microbial growth, formation of tyloses, deposition of materials in the lumen of xylem vessels and the presence of air emboli in the vascular system. Such obstructions may restrict water uptake and its transport towards upwards thus lowering their ornamental value and longevity of cut flowers. Clematis is a very attractive plant material which may be used as cut flower in floral compositions. Nothing is known about the histochemical or cytological nature of xylem blockages occurring in cut stems of this plant. This study shows that in clematis, tyloses are the main source of occlusions, although bacteria and some amorphic substances may also appear inside the vessels. A preservative composed of 200 mg dm−3 8-HQC (8-hydroxyquinolin citrate) and 2% sucrose arrested bacterial development and the growth of tyloses. This information can be helpful in the development of new treatments to improve keeping qualities of cut clematis stems.
PMCID: PMC3417206  PMID: 22919351
9.  Effects of IAA, IBA, NAA, and GA3 on Rooting and Morphological Features of Melissa officinalis L. Stem Cuttings 
The Scientific World Journal  2013;2013:909507.
This study analyzed the potential of producing Melissa officinalis L. using stem cuttings. Four different hormones (IAA, IBA, NAA, and GA3) were applied to the cuttings, with and without buds, in two doses (1000 mg/L and 5000 mg/L), and after 60 days, 10 morphological characteristics of newly generated plants were detected, and a statistical analysis was carried out. The results of the study show that the cuttings with at least one bud must be used in order to produce M. officinalis using stem cuttings. Even though the auxin group hormones (IAA, IBA, and NAA) do not have an apparent effect on rooting percentage, these hormones were detected to affect the morphological characteristics of the newly generated plants, especially root generation. GA3 application has a considerable effect on stem height.
PMCID: PMC3683503  PMID: 23818834
10.  Effect of Naphthalene Acetic Acid on Adventitious Root Development and Associated Physiological Changes in Stem Cutting of Hemarthria compressa 
PLoS ONE  2014;9(3):e90700.
In order to find a way to induce rooting on cuttings of Hemarthria compressa cv. Ya’an under controlled conditions, a project was carried out to study the effect of naphthalene acetic acid (NAA) on rooting in stem cuttings and related physiological changes during the rooting process of Hemarthria compressa. The cuttings were treated with five concentrations of NAA (0, 100, 200 300, 400 mg/l) at three soaking durations (10, 20, 30 minutes), and cuttings without treatment were considered as control. Samples were planted immediately into pots after treatment. IAA-oxidase (IAAO) activity, peroxidase (POD) activity and polyphenol oxidase (PPO) activity were determined after planting. Results showed that NAA had positive effect on rooting at the concentration of 200 mg/l compared to other concentrations at 30 days after planting (DAP). Among the three soaking durations, 20 minutes (min) of 200 mg/l NAA resulted in higher percentages of rooting, larger numbers of adventitious roots and heavier root dry weight per cutting. The lowest IAAO activity was obtained when soaked at 200 mg/l NAA for 20 min soaking duration. This was consistent with the best rooting ability, indicating that the lower IAAO activity, the higher POD activity and PPO activity could be used as an indicator of better rooting ability for whip grass cuttings and might serve as a good marker for rooting ability in cuttings.
PMCID: PMC3942460  PMID: 24595064
11.  Growth of Potato and Control of Pratylenchus penetrans with Oxamyl-treated Seed Pieces in Greenhouse Studies 
Journal of Nematology  1988;20(3):405-409.
Oxamyl was applied to both uncut and cut potato tubers in aqueous solutions of 1,000 to 32,000 μg/ml. Emergence in greenhouse pots was delayed for a day or more after soaking cut tuber pieces in 32,000 μg/ml. After 10 weeks plant growth was greater, relative to the control, when Pratylenchus penetrans-infested soil was planted with cut tubers soaked for 20 minutes in 32,000 μg/ml. Soaking for 40 minutes did not increase nematode control nor affect plant growth. Oxamyl applied to tubers at 1,000 μg/ml reduced the numbers of P. penetrans in the soil by 20% and in the roots by 35%; at 32,000 μg/ml, the numbers of P. penetrans in the soil were reduced by 73-86% and in the roots by 86-97%. The numbers of P. penetrans did not increase in the roots of plants developed from cut tubers soaked in 32,000 μg/ml over a period of 10 weeks, but numbers of lesion nematodes had begun to increase in the soil.
PMCID: PMC2618826  PMID: 19290230
chemical control; root-lesion nematode; Solanum tuberosum; Vydate
12.  Effect of Hirschmanniella caudacrena on the Submersed Aquatic Plants Ceratophyllum demersum and Hydrilla verticillata 
Journal of Nematology  1987;19(4):447-453.
In vitro pathogenicity tests demonstrated that Hirschmanniella caudacrena is pathogenic to Ceratophyllum demersum (coontail). Symptoms were chlorotic tissue, deformed stems, and, finally, death of the plant. Inoculum densities of 500 nematodes per 5-cm-long cutting in a test tube containing 50 ml of water resulted in death and decay of some of the cuttings within 8 weeks; 100 nematodes killed the plants in 12 weeks, and 50 and 25 nematodes killed them in 16 weeks. The lowest inoculum level of 10 nematodes did not seriously affect the plants at 16 weeks when the experiment was terminated. A second test conducted outdoors in glass jars containing 3 liters of water and two cuttings weighing a total of 15 g fresh weight showed damage, but results were not statistically significant. Hydrilla verticillata inoculated with H. caudacrena was not affected seriously.
PMCID: PMC2618666  PMID: 19290169
aquatic plant; biological control; Ceratophyllum demersum; coontail; Hirschmanniella caudacrena; hydrilla; Hydrilla verticillata; pathogenicity test; rice root nematode
13.  Plants use macronutrients accumulated in leaf-cutting ant nests 
Leaf-cutting ants (Atta spp.) are known for their extensive defoliation in neo-tropical forests and savannahs. Debate about the costs and benefits of their activities has been largely dominated by their detrimental effects on agriculture and agroforestry. However, the large accumulation of nutrients and changes in soil properties near their nests might benefit plants growing near them. Here, we test whether trees use nutrients that accumulate in debris piles near, or refuse chambers within, leaf-cutting ant nests. At two tropical sites (a moist tropical forest site in Panama and a savannah site in Brazil), we fed leaves labelled with the stable isotope 15N to two species of leaf-cutting ants (Atta colombica and Atta laevigata) and traced the stable isotope label in plants surrounding the two nests. Thus, we show that plants in both sites access resources associated with Atta nests. In addition, leaf tissue of trees near the nests labelled with 15N had significantly higher calcium concentrations than those of distal, unlabelled conspecifics. It has been documented that calcium is a limiting macronutrient in tropical forests and savannahs. Atta may thus play an important ecological role through their long-distance transport, redistribution and concentration of critical macronutrients.
PMCID: PMC1702380  PMID: 17164194
leaf-cutting ants; Atta; nitrogen isotopes; nutrients; tropical forests; savannahs
14.  Off-season biogenic volatile organic compound emissions from heath mesocosms: responses to vegetation cutting 
Biogenic volatile organic compounds (BVOCs) affect both atmospheric processes and ecological interactions. Our primary aim was to differentiate between BVOC emissions from above- and belowground plant parts and heath soil outside the growing season. The second aim was to assess emissions from herbivory, mimicked by cutting the plants. Mesocosms from a temperate Deschampsia flexuosa-dominated heath ecosystem and a subarctic mixed heath ecosystem were either left intact, the aboveground vegetation was cut, or all plant parts (including roots) were removed. For 3–5 weeks, BVOC emissions were measured in growth chambers by an enclosure method using gas chromatography-mass spectrometry. CO2 exchange, soil microbial biomass, and soil carbon and nitrogen concentrations were also analyzed. Vegetation cutting increased BVOC emissions by more than 20-fold, and the induced compounds were mainly eight-carbon compounds and sesquiterpenes. In the Deschampsia heath, the overall low BVOC emissions originated mainly from soil. In the mixed heath, root, and soil emissions were negligible. Net BVOC emissions from roots and soil of these well-drained heaths do not significantly contribute to ecosystem emissions, at least outside the growing season. If insect outbreaks become more frequent with climate change, ecosystem BVOC emissions will periodically increase due to herbivory.
PMCID: PMC3744039  PMID: 23966983
induced volatiles; BVOC; sesquiterpenes; soil; plant wounding; grazing; Deschampsia flexuosa; arctic
15.  Endophytic fungi reduce leaf-cutting ant damage to seedlings 
Biology Letters  2010;7(1):30-32.
Our study examines how the mutualism between Atta colombica leaf-cutting ants and their cultivated fungus is influenced by the presence of diverse foliar endophytic fungi (endophytes) at high densities in tropical leaf tissues. We conducted laboratory choice trials in which ant colonies chose between Cordia alliodora seedlings with high (Ehigh) or low (Elow) densities of endophytes. The Ehigh seedlings contained 5.5 times higher endophyte content and a greater diversity of fungal morphospecies than the Elow treatment, and endophyte content was not correlated with leaf toughness or thickness. Leaf-cutting ants cut over 2.5 times the leaf area from Elow relative to Ehigh seedlings and had a tendency to recruit more ants to Elow plants. Our findings suggest that leaf-cutting ants may incur costs from cutting and processing leaves with high endophyte loads, which could impact Neotropical forests by causing variable damage rates within plant communities.
PMCID: PMC3030869  PMID: 20610420
Attini; herbivory; symbiosis; endophytes; mutualism
16.  Novel Thylakoid Membrane GreenCut Protein CPLD38 Impacts Accumulation of the Cytochrome b6f Complex and Associated Regulatory Processes* 
The Journal of Biological Chemistry  2013;288(10):7024-7036.
Background: The GreenCut is a group of ∼600 green-lineage-specific proteins hypothetically involved in photosynthesis.
Results: A Chlamydomonas reinhardtii mutant disrupted for GreenCut gene CPLD38 has a marked reduction in cytochrome b6f and an increase in chlororespiration.
Conclusion: CPLD38 is essential for accumulating cytochrome b6f and balancing chlororespiration and photosynthesis.
Significance: This analysis demonstrates the importance of a GreenCut protein in photosynthesis.
Based on previous comparative genomic analyses, a set of nearly 600 polypeptides was identified that is present in green algae and flowering and nonflowering plants but is not present (or is highly diverged) in nonphotosynthetic organisms. The gene encoding one of these “GreenCut” proteins, CPLD38, is in the same operon as ndhL in most cyanobacteria; the NdhL protein is part of a complex essential for cyanobacterial respiration. A cpld38 mutant of Chlamydomonas reinhardtii does not grow on minimal medium, is high light-sensitive under photoheterotrophic conditions, has lower accumulation of photosynthetic complexes, reduced photosynthetic electron flow to P700+, and reduced photochemical efficiency of photosystem II (ΦPSII); these phenotypes are rescued by a wild-type copy of CPLD38. Single turnover flash experiments and biochemical analyses demonstrated that cytochrome b6f function was severely compromised, and the levels of transcripts and polypeptide subunits of the cytochrome b6f complex were also significantly lower in the cpld38 mutant. Furthermore, subunits of the cytochrome b6f complex in mutant cells turned over much more rapidly than in wild-type cells. Interestingly, PTOX2 and NDA2, two major proteins involved in chlororespiration, were more than 5-fold higher in mutants relative to wild-type cells, suggesting a shift in the cpld38 mutant from photosynthesis toward chlororespiratory metabolism, which is supported by experiments that quantify the reduction state of the plastoquinone pool. Together, these findings support the hypothesis that CPLD38 impacts the stability of the cytochrome b6f complex and possibly plays a role in balancing redox inputs to the quinone pool from photosynthesis and chlororespiration.
PMCID: PMC3591612  PMID: 23303190
Algae; Chlamydomonas; Chloroplast; Electron Transport; Genomics; Photosynthesis; Plant Physiology; Comparative Genomics; Cytochrome b6f Complex; Chlororespiration
17.  Do Leaf Cutting Ants Cut Undetected? Testing the Effect of Ant-Induced Plant Defences on Foraging Decisions in Atta colombica 
PLoS ONE  2011;6(7):e22340.
Leaf-cutting ants (LCAs) are polyphagous, yet highly selective herbivores. The factors that govern their selection of food plants, however, remain poorly understood. We hypothesized that the induction of anti-herbivore defences by attacked food plants, which are toxic to either ants or their mutualistic fungus, should significantly affect the ants' foraging behaviour. To test this “induced defence hypothesis,” we used lima bean (Phaseolus lunatus), a plant that emits many volatile organic compounds (VOCs) upon herbivore attack with known anti-fungal or ant-repellent effects. Our results provide three important insights into the foraging ecology of LCAs. First, leaf-cutting by Atta ants can induce plant defences: Lima bean plants that were repeatedly exposed to foraging workers of Atta colombica over a period of three days emitted significantly more VOCs than undamaged control plants. Second, the level to which a plant has induced its anti-herbivore defences can affect the LCAs' foraging behaviour: In dual choice bioassays, foragers discriminated control plants from plants that have been damaged mechanically or by LCAs 24 h ago. In contrast, strong induction levels of plants after treatment with the plant hormone jasmonic acid or three days of LCA feeding strongly repelled LCA foragers relative to undamaged control plants. Third, the LCA-specific mode of damaging leaves allows them to remove larger quantities of leaf material before being recognized by the plant: While leaf loss of approximately 15% due to a chewing herbivore (coccinelid beetle) was sufficient to significantly increase VOC emission levels after 24 h, the removal of even 20% of a plant's leaf area within 20 min by LCAs did not affect its VOC emission rate after 24 h. Taken together, our results support the “induced defence hypothesis” and provide first empirical evidence that the foraging behaviour of LCAs is affected by the induction of plant defence responses.
PMCID: PMC3140513  PMID: 21799831
18.  Effect of Plant Age on Resistance of Alfalfa to Meloidogyne hapla 
Journal of Nematology  1972;4(2):87-90.
Meloidogyne hapla-resistant plants grown from cuttings and inoculated with M. hapla larvae were free of galls. However, 35 to 48% of the seedling intercross progeny of resistant genotypes that were inoculated in the germinated seed stage were galled. There was an inverse relationship between the age of plants grown from seed and the percentage of plants galled by M. hapla; the older the plants at inoculation, the greater the percentage of gall-free plants. The per cent of galled plants was significantly reduced when galled roots were removed and plants reinoculated. Reproduction of M. hapla on galled progeny of resistant plants was significantly less than that on susceptible plants. There were no differences in nematode reproduction on galled progeny of resistant plants, regardless of age at time of inoculation. An in,ease in inoculum levels from 100 to 10,000 M. hapla larvae did not affect resistance or susceptility. There was a direct correlation between galling of inoculated seedlings of resistant progeny and temperature; inoculated 8-week-old cuttings of resistant plants were galled only at 32 C.
PMCID: PMC2619930  PMID: 19319252
selections; temperature
19.  Adhesion of Bacteroides succinogenes in pure culture and in the presence of Ruminococcus flavefaciens to cell walls in leaves of perennial ryegrass (Lolium perenne). 
Bacteroides succinogenes and Ruminococcus flavefaciens are two of the most important cellulolytic bacteria in the rumen. Adhesion of B. succinogenes in pure culture, and in mixed culture with R. flavefaciens, to the various types of cell walls in sections of perennial ryegrass (Lolium perenne L. cultivar S24) leaves was examined by transmission and scanning electron microscopy. B. succinogenes adhered to the cut edges of most plant cell walls except those of the meta- and protoxylem. It also adhered, though in much smaller numbers, to the uncut surfaces of mesophyll, epidermal, and phloem cell walls. In mixed culture, both species adhered in significant numbers to the cut edges of most types of plant cell wall, but R. flavefaciens predominated on the epidermis, phloem, and sclerenchyma cell walls. B. succinogenes predominated on the cut edges and on the uncut surfaces of the mesophyll cell walls, and its ability to adhere to uncut surfaces of other cell walls was not affected by the presence of the ruminococcus. Both organisms rapidly digested the epidermal, mesophyll, and phloem cell walls. Zones of digestion were observed around bacteria of both species when attached to the lignified cell walls of the sclerenchyma, but not when attached to the lignified xylem vessels.
PMCID: PMC243001  PMID: 567035
20.  Microscopic and Molecular Studies of the Diversity of Free-Living Protozoa in Meat-Cutting Plants▿  
Applied and Environmental Microbiology  2008;74(18):5741-5749.
The diversity of free-living protozoa in five meat-cutting plants was determined. Light microscopy after enrichment culturing was combined with sequencing of PCR-amplified, denaturing gradient gel electrophoresis (DGGE)-separated 18S rRNA gene fragments, which was used as a fast screening method. The general results of the survey showed that a protozoan community of amoebae, ciliates, and flagellates was present in all of the plants. Protozoa were detected mainly in floor drains, in standing water on the floor, on soiled bars of cutting tables, on plastic pallets, and in out-of-use hot water knife sanitizers, but they were also detected on surfaces which come into direct contact with meat, such as conveyer belts, working surfaces of cutting tables, and needles of a meat tenderizer. After 7 days of incubation at refrigerator temperature, protozoa were detected in about one-half of the enrichment cultures. Based on microscopic observations, 61 morphospecies were found, and Bodo saltans, Bodo spp., Epistylis spp., Glaucoma scintillans, Petalomonas spp., Prodiscophrya collini, and Vannella sp. were the most frequently encountered identified organisms. Sequencing of DGGE bands resulted in identification of a total of 49 phylotypes, including representatives of the Amoebozoa, Chromalveolata, Excavata, Opisthokonta, and Rhizaria. Sequences of small heterotrophic flagellates were affiliated mainly with the Alveolata (Apicomplexa), Stramenopiles (Chrysophyceae), and Rhizaria (Cercozoa). This survey showed that there is high protozoan species richness in meat-cutting plants and that the species included species related to known hosts of food-borne pathogens.
PMCID: PMC2547033  PMID: 18641165
21.  Stem Nematode-Fusarium Wilt Complex in Alfalfa as Related to Irrigation Management at Harvest Time 
Journal of Nematology  1992;24(2):315-320.
A high moisture level in the top 10 cm of soil at time of cutting of alfalfa increased the incidence of plant mortality and Fusarium wilt in soil infested with Ditylenchus dipsaci and Fusarium oxysporum f. sp. medicaginis in greenhouse and field microplot studies. Ranger alfalfa, susceptible to both D. dipsaci and F. oxysporum f. sp. medicaginis, was less persistent than Moapa 69 (nematode susceptible and Fusarium wilt resistant) and Lahontan alfalfa (nematode resistant with low Fusarium wilt resistance). In the greenhouse, the persistence of Ranger, Moapa 69, and Lahontan alfalfa plants was 46%, 64%, and 67% respectively, in nematode + fungus infested soil at high soil moisture at time of cutting. This compared to 74%, 84%, and 73% persistence of Ranger, Moapa 69, and Lahontan, respectively, at low soil moisture at time of cutting. Shoot weights as a percentage of uninoculated controls at the high soil moisture level were 38%, 40%, and 71% for Ranger, Moapa 69, and Lahontan, respectively. Low soil moisture at time of cutting negated the effect D. dipsaci on plant persistence and growth of subsequent cuttings, and reduced Fusarium wilt of plants in the nematode-fungus treatment; shoot weights were 75%, 90%, and 74% of uninoculated controls for Ranger, Moapa 69, and Lahontan. Similar results were obtained in the field microplot study, and stand persistence and shoot weights were less in nematode + fungus-infested soil at the high soil-moisture level (early irrigation) than at the low soil-moisture level (late irrigation).
PMCID: PMC2619274  PMID: 19283002
alfalfa; Ditylenchus dipsaci; Fusarium oxysporum f. sp. medicaginis; interaction; irrigation timing; Medicago sativa; mortality; nematode; soil moisture; suppression
22.  Influence of Microgravity Environment on Root Growth, Soluble Sugars, and Starch Concentration of Sweetpotato Stem Cuttings 
Because sweetpotato [Ipomoea batatas (L.) Lam.] stem cuttings regenerate very easily and quickly, a study of their early growth and development in microgravity could be useful to an understanding of morphological changes that might occur under such conditions for crops that are propagated vegetatively. An experiment was conducted aboard a U.S. Space Shuttle to investigate the impact of microgravity on root growth, distribution of amyloplasts in the root cells, and on the concentration of soluble sugars and starch in the stems of sweetpotatoes. Twelve stem cuttings of ‘Whatley/Loretan’ sweetpotato (5 cm long) with three to four nodes were grown in each of two plant growth units filled with a nutrient agarose medium impregnated with a half-strength Hoagland solution. One plant growth unit was flown on Space Shuttle Colombia for 5 days, whereas the other remained on the ground as a control. The cuttings were received within 2 h postflight and, along with ground controls, processed in ≈45 min. Adventitious roots were counted, measured, and fixed for electron microscopy and stems frozen for starch and sugar assays. Air samples were collected from the headspace of each plant growth unit for postflight determination of carbon dioxide, oxygen, and ethylene levels. All stem cuttings produced adventitious roots and growth was quite vigorous in both ground-based and flight samples and, except for a slight browning of some root tips in the flight samples, all stem cuttings appeared normal. The roots on the flight cuttings tended to grow in random directions. Also, stem cuttings grown in microgravity had more roots and greater total root length than ground-based controls. Amyloplasts in root cap cells of ground-based controls were evenly sedimented toward one end compared with a more random distribution in the flight samples. The concentration of soluble sugars, glucose, fructose, and sucrose and total starch concentration were all substantially greater in the stems of flight samples than those found in the ground-based samples. Carbon dioxide levels were 50% greater and oxygen marginally lower in the flight plants, whereas ethylene levels were similar and averaged less than 10 nL·L −1. Despite the greater accumulation of carbohydrates in the stems, and greater root growth in the flight cuttings, overall results showed minimal differences in cell development between space flight and ground-based tissues. This suggests that the space flight environment did not adversely impact sweetpotato metabolism and that vegetative cuttings should be an acceptable approach for propagating sweetpotato plants for space applications.
PMCID: PMC2827937  PMID: 20186286
hypoxia; Ipomea batata; Phytagel; carbohydrate metabolism; bioregenerative life support
23.  The role of plant traits and their plasticity in the response of pasture grasses to nutrients and cutting frequency 
Annals of Botany  2010;105(6):957-965.
Background and Aims
Although plant functional traits (PFTs) appear to be important indicators of species' responses to land use changes, there is no clear understanding of how the variations in traits and their plasticity determine variations in species performance. This study investigated the role of functional shoot traits and their plasticity for variation in above-ground net primary productivity (ANPP) due to changes in N supply and in cutting frequency for 13 native perennial C3 grass species.
Monocultures of the grass species were grown in a fully factorial block design combining plant species, cutting frequency and N supply as factors.
Key Results
Four major trait associations were obtained by reducing the dimensions of 14 PFTs with a principal component analysis (PCA).Variations in species' productivity in response to an increase in cutting frequency was mainly explained by traits linked to the first PCA axis, opposing high plant stature from lower shoot cellulose and lignin contents and high leaf N content. Variation in species productivity in response to change in N supply was mainly explained by a set of predictor variables combining traits (average flowering date) and a trait's plasticity (tiller density per unit land area and leaf dry matter content, i.e. mg dry matter g fresh mass−1). These traits involved are linked to the second PCA axis (‘nutrient acquisition–conservation’), which opposes distinct strategies based on response to nutrient supply.
Variations in ANPP of species in response to an increase in cutting frequency and a decrease in N supply are controlled by a group of traits, rather than by one individual trait. Incorporating plasticity of the individual traits into these trait combinations was the key to explaining species' productivity responses, accounting for up to 89 % of the total variability in response to the changes in N supply.
PMCID: PMC2876009  PMID: 20354073
C3 grasses; cutting; grassland; leaf traits; nitrogen; primary productivity; species strategy; trait plasticity
24.  Foraging of Psilocybe basidiocarps by the leaf-cutting ant Acromyrmex lobicornis in Santa Fé, Argentina 
SpringerPlus  2013;2:254.
It is generally accepted that material collected by leaf-cutting ants of the genus Acromyrmex consists solely of plant matter, which is used in the nest as substrate for a symbiotic fungus providing nutrition to the ants. There is only one previous report of any leaf-cutting ant foraging directly on fungal basidiocarps.
Basidiocarps of Psilocybe coprophila growing on cow dung were actively collected by workers of Acromyrmex lobicornis in Santa Fé province, Argentina. During this behaviour the ants displayed typical signals of recognition and continuously recruited other foragers to the task. Basidiocarps of different stages of maturity were being transported into the nest by particular groups of workers, while other workers collected plant material.
The collection of mature basidiocarps with viable spores by leaf-cutting ants in nature adds substance to theories relating to the origin of fungiculture in these highly specialized social insects.
Electronic supplementary material
The online version of this article (doi:10.1186/2193-1801-2-254) contains supplementary material, which is available to authorized users.
PMCID: PMC3679412  PMID: 23772357
Acromyrmex lobicornis; Basidiocarps; Coprophilous fungus; Deconica coprophila; Forage behaviour; Leaf-cutting ants; Psilocybe coprophila
25.  Towards a functional–structural plant model of cut-rose: simulation of light environment, light absorption, photosynthesis and interference with the plant structure 
Annals of Botany  2011;108(6):1121-1134.
Background and Aims
The production system of cut-rose (Rosa × hybrida) involves a complex combination of plant material, management practice and environment. Plant structure is determined by bud break and shoot development while having an effect on local light climate. The aim of the present study is to cover selected aspects of the cut-rose system using functional–structural plant modelling (FSPM), in order to better understand processes contributing to produce quality and quantity.
The model describes the production system in three dimensions, including a virtual greenhouse environment with the crop, light sources (diffuse and direct sun light and lamps) and photosynthetically active radiation (PAR) sensors. The crop model is designed as a multiscaled FSPM with plant organs (axillary buds, leaves, internodes, flowers) as basic units, and local light interception and photosynthesis within each leaf. A Monte-Carlo light model was used to compute the local light climate for leaf photosynthesis, the latter described using a biochemical rate model.
Key Results
The model was able to reproduce PAR measurements taken at different canopy positions, different times of the day and different light regimes. Simulated incident and absorbed PAR as well as net assimilation rate in upright and bent shoots showed characteristic spatial and diurnal dynamics for different common cultivation scenarios.
The model of cut-rose presented allowed the creation of a range of initial structures thanks to interactive rules for pruning, cutting and bending. These static structures can be regarded as departure points for the dynamic simulation of production of flower canes. Furthermore, the model was able to predict local (per leaf) light absorption and photosynthesis. It can be used to investigate the physiology of ornamental plants, and provide support for the decisions of growers and consultants.
PMCID: PMC3189845  PMID: 21856634
Cut-rose; Rosa × hybrida; bud break; light distribution; interactive modelling; functional–structural plant model; FSPM; L-system; virtual PAR sensor

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