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"Graham, James H"
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Little evidence that farmers should consider abundance or diversity of arbuscular mycorrhizal fungi when managing crops
2018
Arbuscular mycorrhizal fungi (AMF) are ubiquitous in agroecosystems and often stated to be critical for crop yield and agroecosystem sustainability. However, should farmers modify management to enhance the abundance and diversity of AMF? We address this question with a focus on field experiments that manipulated colonisation by indigenous AMF and report crop yield, or investigated community structure and diversity of AMF. We find that the literature presents an overly optimistic view of the importance of AMF in crop yield due, in part, to flawed methodology in field experiments. A small body of rigorous research only sometimes reports a positive impact of high colonisation on crop yield, even under phosphorus limitation. We suggest that studies vary due to the interaction of environment and genotype (crop and mycorrhizal fungal). We also find that the literature can be overly pessimistic about the impact of some common agricultural practices on mycorrhizal fungal communities and that interactions between AMF and soil microbes are complex and poorly understood. We provide a template for future field experiments and a list of research priorities, including phosphorus-efficient agroecosystems. However, we conclude that management of AMF by farmers will not be warranted until benefits are demonstrated at the field scale under prescribed agronomic management.
Journal Article
Presence of Extracellular DNA during Biofilm Formation by Xanthomonas citri subsp. citri Strains with Different Host Range
by
Sena-Vélez, Marta
,
Redondo, Cristina
,
Graham, James H.
in
Antibiotics
,
Bacteria
,
Bacteriology
2016
Xanthomonas citri subsp. citri (Xcc) A strain causes citrus bacterial canker, a serious leaf, fruit and stem spotting disease of several Citrus species. X. alfalfae subsp. citrumelonis (Xac) is the cause of citrus bacterial spot, a minor disease of citrus nursery plants and X. campestris pv. campestris (Xc) is a systemic pathogen that causes black rot of cabbage. Xanthomonas spp. form biofilms in planta that facilitate the host infection process. Herein, the role of extracellular DNA (eDNA) was evaluated in the formation and stabilization of the biofilm matrix at different stages of biofilm development. Fluorescence and light microscopy, as well as DNAse treatments, were used to determine the presence of eDNA in biofilms and bacterial cultures. DNAse treatments of Xcc strains and Xac reduced biofilm formation at the initial stage of development, as well as disrupted preformed biofilm. By comparison, no significant effect of the DNAse was detected for biofilm formation by Xc. DNAse effects on biofilm formation or disruption varied among Xcc strains and Xanthomonas species which suggest different roles for eDNA. Variation in the structure of fibers containing eDNA in biofilms, bacterial cultures, and in twitching motility was also visualized by microscopy. The proposed roles for eDNA are as an adhesin in the early stages of biofilm formation, as an structural component of mature bacterial aggregates, and twitching motility structures.
Journal Article
Plant mitochondrial introns as genetic markers - conservation and variation
2023
Plant genomes are comprised of nuclear, plastid and mitochondrial components characterized by different patterns of inheritance and evolution. Genetic markers from the three genomes provide complementary tools for investigations of inheritance, genetic relationships and phenotypic contributions. Plant mitochondrial genomes are challenging for universal marker development because they are highly variable in terms of size, gene order and intergenic sequences and highly conserved with respect to protein-coding sequences. PCR amplification of introns with primers that anneal to conserved, flanking exons is effective for the development of polymorphic nuclear genome markers. The potential for plant mitochondrial intron polymorphisms to distinguish between congeneric species or intraspecific varieties has not been systematically investigated and is possibly constrained by requirements for intron secondary structure and interactions with co-evolved organelle intron splicing factors. To explore the potential for broadly applicable plant mitochondrial intron markers, PCR primer sets based upon conserved sequences flanking 11 introns common to seven angiosperm species were tested across a range of plant orders. PCR-amplified introns were screened for indel polymorphisms among a group of cross-compatible Citrus species and relatives; two Raphanus sativus mitotypes; representatives of the two Phaseolus vulgaris gene pools; and congeneric pairs of Cynodon , Cenchrus , Solanum , and Vaccinium species. All introns were successfully amplified from each plant entry. Length polymorphisms distinguishable by gel electrophoresis were common among genera but infrequent within genera. Sequencing of three introns amplified from 16 entries identified additional short indel polymorphisms and nucleotide substitutions that separated Citrus , Cynodon , Cenchrus and Vaccinium congeners, but failed to distinguish Solanum congeners or representatives of the Phaseolus vulgaris major gene pools. The ability of primer sets to amplify a wider range of plant species’ introns and the presence of intron polymorphisms that distinguish congeners was confirmed by in silico analysis. While mitochondrial intron variation is limited in comparison to nuclear introns, these exon-based primer sets provide robust tools for the amplification of mitochondrial introns across a wide range of plant species wherein useful polymorphisms can be identified.
Journal Article
Nutrient Status and Root Density of Huanglongbing-Affected Trees: Consequences of Irrigation Water Bicarbonate and Soil pH Mitigation with Acidification
2019
The Candidatus Liberibacter asiaticus bacterium, associated with Huanglongbing (HLB) disease of citrus trees, moves downward in the phloem and infects the roots soon after transmission by the Asian Citrus Psyllid (Diaphorina citri) vector into shoots. Before canopy symptoms appear, 30–50% of the roots are damaged. Without aggressive management to reduce abiotic and biotic stress, root loss increases to 70–80%. An extensive survey of HLB-affected groves in central and south-central Florida indicated that a greater decline in fibrous root health as well as a greater expression of HLB symptoms is observed where irrigation water is high in bicarbonates (>100 mg L−1) and soil pH is >6.5. Over three seasons of survey, acidification of irrigation water in the central and south-central citrus growing regions of Florida reduced the decline in root density associated with HLB. Irrigation water treatment with sulfuric acid and soil amendment with elemental sulfur for 36 months to establish a soil pH range from 4.0 to 7.0 increased root growth, soil nutrient availability, and the uptake of Ca, Mg, Mn, and Zn in response to a gradual reduction in soil pH in young and mature Valencia orange groves on Swingle citrumelo rootstock. The reduction in soil pH increased yield and soluble solids in fruit and so would improve citrus production.
Journal Article
The continuum concept remains a useful framework for studying mycorrhizal functioning
by
Johnson, Nancy Collins
,
Graham, James H.
in
Agronomy. Soil science and plant productions
,
Animal, plant and microbial ecology
,
Biogeochemistry
2013
Background Recent studies have questioned the validity of the mutualism-parasitism continuum of mycorrhizal function. This paper re-evaluates the continuum model and analyzes these concerns. Scope and Conclusions Three insights arise from this analysis. First, the continuum model defines mycorrhizal function as an emergent property of complex interactions. The model identifies resource trade and symbiotic control as key determinants of the costs and benefits of the symbiosis for plants and fungi, and the interaction of these factors with the environment ultimately controls mycorrhizal function. Second, analysis of carbon costs and phosphorus benefits is too narrow a focus to accurately predict mycorrhizal function. Analysis of plant and fungal fitness responses in ecologically and evolutionarily relevant systems are required to elucidate the full range of nutritional and non-nutritional factors embodied within mycorrhizal functioning. Finally, the definition of the term 'parasitism' has evolved. Some fields of science maintain the original definition of a nutritional relationship between host and parasite while other fields define it as a +/- fitness relationship. This has generated debate about whether the continuum of mycorrhizal functioning should properly be called a positive-negative response continuum or a mutualism-parasitism continuum. This controversy about semantics should be resolved, but it does not overturn the continuum concept.
Journal Article
Chemotactic Responses of Xanthomonas with Different Host Ranges
2022
Xanthomonas citri pv. citri (Xcc) (X. citri subsp. citri) type A is the causal agent of citrus bacterial canker (CBC) on most Citrus spp. and close relatives. Two narrow-host-range strains of Xcc, Aw and A*, from Florida and Southwest Asia, respectively, infect only Mexican lime (Citrus aurantifolia) and alemow (C. macrophylla). In the initial stage of infection, these xanthomonads enter via stomata to reach the apoplast. Herein, we investigated the differences in chemotactic responses for wide and narrow-host-range strains of Xcc A, X. euvesicatoria pv. citrumelonis (X. alfalfae subsp. citrumelonis), the causal agent of citrus bacterial spot, and X. campestris pv. campestris, the crucifer black rot pathogen. These strains of Xanthomonas were compared for carbon source use, the chemotactic responses toward carbon compounds, chemotaxis sensor content, and responses to apoplastic fluids from Citrus spp. and Chinese cabbage (Brassica pekinensis). Different chemotactic responses occurred for carbon sources and apoplastic fluids, depending on the Xanthomonas strain and the host plant from which the apoplastic fluid was derived. Differential chemotactic responses to carbon sources and citrus apoplasts suggest that these Xanthomonas strains sense host-specific signals that facilitate their location and entry of stomatal openings or wounds.
Journal Article
Phospholipase D (PLD) Response to Water Stress in Citrus Roots and Leaves
2020
The effects of drought stress on phospholipase D (PLD) gene expression and enzymatic activity were investigated in ‘Pineapple’ sweet orange. PLDs are tissue-specific, with overlapping functions, and in response to stress they may interact with ABA signaling. Tissue specificity for expression of PLDs and their regulation are unknown in citrus. To assess PLD response to water stress and gene expression/regulation in citrus, we subjected potted seedlings to increasing levels of soil drought. Evapotranspirational demand (ET) was estimated by measuring weight loss in pots, and water stress was further assessed by measuring ABA content. Three treatments were performed over a 3-week period: (a) Control treatment without drought stress (100% of daily ET); (b) mild water stress (50% ET); and (c) severe water stress (0% ET). ABA content increased during drought stress in both roots and leaves, being higher in leaves than in roots by the end of the experiment for any stress condition assayed. PLD enzymatic activity was monitored and expression of five PLD genes was studied. PLD activity increased linearly over time in response to increased soil drought and was around three times higher in roots than in leaves. PLD activation occurred initially in roots and then in leaves. PLD gene expression in response to stress soil drought differed between roots and leaves. These results show the potential of PLD as a suitable indicator of stress severity in citrus.
Journal Article
Novel Plastid-Nuclear Genome Combinations Enhance Resistance to Citrus Canker in Cybrid Grapefruit
2019
Host disease resistance is the most desirable strategy for control of citrus canker, a disease caused by a gram-negative bacterium
subsp.
. However, no resistant commercial citrus cultivar has been identified. Cybridization, a somatic hybridization approach that combines the organelle and nuclear genomes from different species, was used to create cybrids between citrus canker resistant 'Meiwa' kumquat (
Swingle snym.
Thunb.) and susceptible grapefruit (
Macfad) cultivars. From these fusions, cybrids with grapefruit nucleus, kumquat mitochondria and kumquat chloroplasts and cybrids with grapefruit nucleus, kumquat mitochondria and grapefruit chloroplasts were generated. These cybrids showed a range of citrus canker response, but all cybrids with kumquat chloroplasts had a significantly lower number of lesions and lower
subsp.
populations than the grapefruit controls. Cybrids with grapefruit chloroplasts had a significantly higher number of lesions than those with kumquat chloroplasts. To understand the role of chloroplasts in the cybrid disease defense, quantitative PCR was performed on both cybrid types and their parents to examine changes in gene expression during
subsp.
infection. The results revealed chloroplast influences on nuclear gene expression, since isonuclear cybrids and 'Marsh' grapefruit had different gene expression profiles. In addition, only genotypes with kumquat chloroplasts showed an early up-regulation of reactive oxygen species genes upon
subsp.
infection. These cybrids have the potential to enhance citrus canker resistance in commercial grapefruit orchards. They also serve as models for understanding the contribution of chloroplasts to plant disease response and raise the question of whether other alien chloroplast genotypes would condition similar results.
Journal Article
Negative plant–soil feedbacks are stronger in agricultural habitats than in forest fragments in the tropical Andes
by
Mangan, Scott A.
,
Kitajima, Kaoru
,
Pizano, Camila
in
Agricultural land
,
Andes region
,
arbuscular mycorrhizal fungi
2019
There is now strong evidence suggesting that interactions between plants and their species-specific antagonistic microbes can maintain native plant community diversity. In contrast, the decay in diversity in plant communities invaded by nonnative plant species might be caused by weakening negative feedback strengths, perhaps because of the increased relative importance of plant mutualists such as arbuscular mycorrhizal fungi (AMF). Although the vast majority of studies examining plant–soil feedbacks have been conducted in a single habitat type, there are fewer studies that have tested how the strength and direction of these feedbacks change across habitats with differing dominating plants. In a fragmented montane agricultural system in Colombia, we experimentally teased apart the relative importance of AMF and non-AMF microbes (a microbial filtrate) to the strength and direction of feedbacks in both native and nonnative plant species. We hypothesized that native tree species of forest fragments would exhibit stronger negative feedbacks with a microbial filtrate that likely contained pathogens than with AMF alone, whereas nonnative plant species, especially a highly invasive dominant grass, would exhibit overall weaker negative feedbacks or even positive feedbacks regardless of the microbial type. We reciprocally inoculated each of 10 plant species separately with either the AMF community or the microbial filtrate originating from their own conspecifics, or with the AMF or microbial filtrate originating from each of the other nine heterospecific plant species. Overall, we found that the strength of negative feedback mediated by the filtrate was much stronger than feedbacks mediated by AMF. Surprisingly, we found that the two nonnative species, Urochloa brizantha and Coffea arabica, experienced stronger negative feedbacks with microbial filtrate than did the native forest tree species, suggesting that species-specific antagonistic microbes accumulate when a single host species dominates, as is the case in agricultural habitats. However, negative feedback between forest trees and agricultural species suggests that soil community dynamics may contribute to the re-establishment of native species into abandoned agricultural lands. Furthermore, our finding of no negative feedbacks among trees in forest fragments may be due to a loss in diversity of those microbes that drive diversity-maintaining processes in intact tropical forests.
Journal Article