scholarly journals Efficient Fluorescence-Based Localization Technique for Tracking Endophytes Route in Host-Plants Colonization

2021 ◽  
Author(s):  
Christine Andeme Ondzighi-Assoume ◽  
Bandana Bhusal ◽  
Adam Traore ◽  
Wilson K Ouma ◽  
Margaret Mmbaga ◽  
...  

Bacterial isolates that enhance plant growth and suppress plant pathogens growth are essential tools for reducing pesticide applications in plant production systems. The objectives of this study were to develop a reliable fluorescence-based technique for labeling bacterial isolates selected as biological control agents (BCAs) to allow their direct tracking in the host-plant interactions, understand the BCA localization within their host plants, and the route of plant colonization. Objectives were achieved by developing competent BCAs transformed with two plasmids, pBSU101 and pANIC-10A, containing reporter genes eGFP and pporRFP, respectively. Our results revealed that the plasmid-mediated transformation efficiencies of antibiotic-resistant competent BCAs identified as PSL, IMC8, and PS were up 84%. Fluorescent BCA-tagged reporter genes were associated with roots and hypocotyls but not with leaves or stems and were confirmed by fluoresence microscopy and PCR analyses in colonized Arabidopsis and sorghum. This fluorescence-based technique's high resolution and reproducibility make it a platform-independent system that allows tracking of BCAs spatially within plant tissues, enabling assessment of the movement and niches of BCAs within colonized plants. Steps for producing and transforming competent fluorescent BCAs, as well as the inoculation of plants with transformed BCAs, localization, and confirmation of fluorescent BCAs through fluorescence imaging and PCR, are provided in this manuscript. This study features host-plant interactions and subsequently biological and physiological mechanisms implicated in these interactions. The maximum time to complete all the steps of this protocol is approximately three months.

Author(s):  
Marcin W. Zielonka ◽  
Tom W. Pope ◽  
Simon R. Leather

Abstract The carnation tortrix moth, Cacoecimorpha pronubana (Hübner, [1799]) (Lepidoptera: Tortricidae), is one of the most economically important insect species affecting the horticultural industry in the UK. The larvae consume foliage, flowers or fruits, and/or rolls leaves together with silken threads, negatively affecting the growth and/or aesthetics of the crop. In order to understand the polyphagous behaviour of this species within an ornamental crop habitat, we hypothesized that different host plant species affect its life history traits differently. This study investigated the effects of the host plant species on larval and pupal durations and sizes, and fecundity (the number of eggs and the number and size of egg clutches). At 20°C, 60% RH and a 16L:8D photoperiod larvae developed 10, 14, 20 and 36 days faster when reared on Christmas berry, Photinia (Rosaceae), than on cherry laurel, Prunus laurocerasus (Rosaceae), New Zealand broadleaf, Griselinia littoralis (Griseliniaceae), Mexican orange, Choisya ternata (Rutaceae), and firethorn, Pyracantha angustifolia (Rosaceae), respectively. Female pupae were 23.8 mg heavier than male pupae, and pupal weight was significantly correlated with the duration of larval development. The lowest and the highest mean numbers of eggs were produced by females reared on Pyracantha (41) and Photinia (202), respectively. Clutch size differed significantly among moths reared on different host plants, although the total number of eggs did not differ. This study showed that different ornamental host plants affect the development of C. pronubana differently. Improved understanding of the influence of host plant on the moth's life history parameters measured here will help in determining the economic impact that this species may have within the ornamental plant production environment, and may be used in developing more accurate crop protection methodologies within integrated pest management of this insect.


1995 ◽  
Vol 73 (S1) ◽  
pp. 1275-1283 ◽  
Author(s):  
Shigehito Takenaka

To develop efficient control measures against fungal plant pathogens, the dynamics of host plant colonization during disease development and the interactions among fungi within host plant tissues need to be clarified. These studies require accurate quantitative estimation of specific fungal biomass in plant tissues. This has been approached by direct-microscopic methods, cultural methods, chemical determinations of fungal components, serological methods, and molecular methods. Among these methods, serological and molecular methods provide rapid, specific, and sensitive quantitative measures of fungal biomass in host plant tissues. Therefore, studies on fungal dynamics of host plant colonization using these two methods are presented. Some examples of species interactions among pathogenic fungi within host plants, such as synergism and competition, are reviewed and the usefulness of serological and molecular methods for studies on these interactions is presented. These quantitative methods will provide helpful information for understanding the ecology of plant pathogenic fungi, such as the dynamics of host plant colonization and species interactions. Key words: quantitative methods, fungal biomass, ELISA, PCR, fungal colonization, species interaction.


Nematology ◽  
2004 ◽  
Vol 6 (3) ◽  
pp. 375-387 ◽  
Author(s):  
N. Aileen Ryan ◽  
Peter Jones

AbstractSeventy bacteria, isolated from the rhizosphere of the potato cyst nematode (PCN) host plant, potato, were cultured in the presence and absence of potato root leachate (PRL) and the resultant culture filtrates were analysed for their ability to affect the hatch in vitro of the two PCN species. Of the isolates tested, nine had a significant effect on PCN hatch. Six affected Globodera pallida hatch and three affected G. rostochiensis hatch. Five of the isolates significantly increased hatch only when cultured in the presence of PRL. Three of the isolates decreased PCN hatch significantly in PRL. Only one isolate increased hatch significantly in the absence of PRL. No isolate affected the hatch of both species. Six of the nine isolates that significantly affected PCN hatch had been pre-selected by culturing on PRL. Bacterial isolates from PCN non-hosts (14 from wheat, 17 from sugar beet) were also tested for hatching activity. The principal effect of the hatch-active isolates from the PCN non-host plants was to increase PCN hatch in the presence of PRL. In contrast to the host bacteria results, the isolates from non-host plants affected only G. rostochiensis hatch (three wheat isolates and four sugar beet isolates significantly increased G. rostochiensis hatch); no such isolate affected G. pallida hatch significantly in the presence of PRL. Ten isolates (32%) from non-host plants had the ability to increase significantly the hatch of PCN in the absence of PRL (eight of these affected G. rostochiensis hatch and four affected G. pallida hatch), compared to only one bacterial isolate (1%) from a host plant. The majority of the isolates from non-hosts produced PCN species-specific effects, as with the bacteria isolated from potatoes, although two wheat isolates increased the hatch of both species significantly in the absence of PRL. Of 20 hatch-active bacterial isolates (from all three plants) identified, 70% were Bacillus spp. Other genera identified were Arthrobacter , Acinetobacter and Staphylococcus .


Water ◽  
2019 ◽  
Vol 11 (12) ◽  
pp. 2558 ◽  
Author(s):  
Andrew Ristvey ◽  
Bruk Belayneh ◽  
John Lea-Cox

Water security in ornamental plant production systems is vital for maintaining profitability. Expensive, complicated, or potentially dangerous treatment systems, together with skilled labor, is often necessary to ensure water quality and plant health. Two contrasting commercial ornamental crop production systems in a mesic region are compared, providing insight into the various strategies employed using irrigation-water containment and treatment systems. The first is a greenhouse/outdoor container operation which grows annual ornamental plants throughout the year using irrigation booms, drip emitters, and/or ebb and flow systems depending on the crop, container size, and/or stage of growth. The operation contains and recycles 50–75% of applied water through a system of underground cisterns, using a recycling reservoir and a newly constructed 0.25 ha slow-sand filtration (SSF) unit. Groundwater provides additional water when needed. Water quantity is not a problem in this operation, but disease and water quality issues, including agrochemicals, are of potential concern. The second is a perennial-plant nursery which propagates cuttings and produces field-grown trees and containerized plants. It has a series of containment/recycling reservoirs that capture rainwater and irrigation return water, together with wells of limited output. Water quantity is a more important issue for this nursery, but poor water quality has had some negative economic effects. Irrigation return water is filtered and sanitized with chlorine gas before being applied to plants via overhead and micro-irrigation systems. The agrochemical paclobutrazol was monitored for one year in the first operation and plant pathogens were qualified and quantified over two seasons for both production systems. The two operations employ very different water treatment systems based on their access to water, growing methods, land topography, and capital investment. Each operation has experienced different water quantity and quality vulnerabilities, and has addressed these threats using a variety of technologies and management techniques to reduce their impacts.


2019 ◽  
Author(s):  
Hesham A.Y. Gibriel ◽  
Jinling Li ◽  
Longfu Zhu ◽  
Michael F. Seidl ◽  
Bart P.H.J. Thomma

Originality and significance statementDuring host colonization, plant pathogens secrete molecules that enable host colonization, also known as effector proteins. Here, we show that strains of the fungal plant pathogen Verticillium dahliae that are able to infect the same host plant harbour highly divergent LS effector repertoires. Our study outlines the variability within LS effector gene repertoires of V. dahliae strains, which may allow the various strains to be competitive in the co-evolution with their hosts.SummaryEffectors are proteins secreted by pathogens to support colonization of host plants, often by deregulating host immunity. Effector genes are often localized within dynamic lineage-specific (LS) genomic regions, allowing rapid evolution of effector catalogues. Such localization permits pathogens to be competitive in the co-evolutionary arms races with their hosts. For a broad host-range pathogen such as Verticillium dahliae it is unclear to what extent single members of their total effector repertoires contribute to disease development on multiple hosts. Here, we determined the core and LS effector repertoires of a collection of V. dahliae strains, as well as the ability of these strains to infect a range of plant species comprising tomato, cotton, Nicotiana benthamiana, Arabidopsis, and sunflower to assess whether the presence of particular LS effectors correlates with the ability to infect particular plant species. Surprisingly, we found that V. dahliae strains that are able to infect the same host plant harbor highly divergent LS effector repertoires. Furthermore, we observed differential V. dahliae core effector gene expression between host plants. Our data suggest that different V. dahliae lineages utilise divergent effector catalogs to colonize the same host plant, suggesting considerable redundancy among the activities of effector catalogs between lineages.


2021 ◽  
Vol 21 (3) ◽  
Author(s):  
Maria Virginia Urso-Guimarães ◽  
Ingrid Koch ◽  
Ana Carolina Devides Castello

Abstract: The Midwest region of Brazil possesses large areas dominated by the Cerrado that is poorly known concerning insect gall and their interactions. In this study, we inventory the gall morphology, host plants, and the gall makers from Parque Nacional da Chapada dos Guimarães, Mato Grosso State, in areas of natural vegetation from Cerrado, for the first time. Samplings occurred in two expeditions, July 2012 and January 2013. We characterized 295 morphotypes of insect galls in 140 host plant species, with 89 gall makers; the richest family in host plants was Fabaceae (16.7%), and the species was Protium heptaphyllum (Aubl.) Marchand (Burseraceae, 3.7%). Parque Nacional da Chapada dos Guimarães is the richest Brazilian cerrado area in gall morphotypes (295) and the second in average morphotypes/host plant species (2.1). Additionally, six genera and 38 species are new records as host plants; two of them, Bernardia similis Pax and K.Hoffm and Ormosia macrophylla Benth., are new occurrences for Mato Grosso State, and other two, Vochysia petraea Warm. and Talisia subalbens (Mart.) Radlk. are listed in the Red List of Threatened Species IUCN. This inventory data represents a testimony of insect-plant interactions in a Brazilian Cerrado area that was consumed by an unprecedented fire in the dry season of 2020.


2021 ◽  
Vol 21 (3) ◽  
Author(s):  
Maria Virginia Urso-Guimarães ◽  
Ingrid Koch ◽  
Ana Carolina Devides Castello

Abstract: The Pantanal Biome occupies 20% of the Brazilian territory extending its distribution over two Brazilian States, Mato Grosso and Mato Grosso do Sul. This Biome is one of Brazil's poorly known regions concerning insect gall and their interactions with host plants. In this study, we characterized for the first time the gall morphology, identified host plants and the gall makers from an area of Brazilian wetlands from Mato Grosso State, known as Pantanal Matogrossense. We sampled Pantanal Biome areas in Poconé municipality, along the Transpantaneira Road, Mato Grosso State, Brazil, in two expeditions, July 2012 and January 2013, with a total effort of 2 hours. We characterized 91 morphotypes of insect galls in 54 host plant species; 28 gall makers in 24 host plant species; the richest host plant families are Fabaceae, Myrtaceae, and Sapindaceae. Psidium guineense Sw. is the super host species. This area in Pantanal Matogrossense is the second in the richness of gall morphotypes (N=91) and average morphotypes/plant species (1.7), comparing phytophysiognomies. Additionally, 15 plant species are new record as host in galler-host plant interaction in the world. This number represents 30% of the total of host plant species sampled in Poconé. This inventory is new knowledge to the Pantanal Matogrossense and representing a unique testimony of insect-plant interactions consumed by the unprecedented fire that occurred in Pantanal Biome in the dry season of 2020.


2008 ◽  
Vol 98 (4) ◽  
pp. 333-341 ◽  
Author(s):  
K. Charaabi ◽  
J. Carletto ◽  
P. Chavigny ◽  
M. Marrakchi ◽  
M. Makni ◽  
...  

AbstractThe study of intraspecific variation with respect to host plant utilization in polyphagous insects is crucial for understanding evolutionary patterns of insect-plant interactions. Aphis gossypii (Glover) is a cosmopolitan and extremely polyphagous aphid species. If host plant species or families constitute selective regimes to these aphids, genetic differentiation and host associated adaptation may occur. In this study, we describe the genetic structure of A. gossypii collected in six localities in Tunisia on different vegetable crops, on citrus trees and on Hibiscus. The aim was to determine if the aphid populations are structured in relation to the host plants and if such differentiation is consistent among localities. The genetic variability of A. gossypii samples was examined at eight microsatellite loci. We identified only 11 multilocus genotypes among 559 individuals. Significant deviations from Hardy-Weinberg equilibrium, linkage disequilibria and absence of recombinant genotypes, confirmed that A. gossypii reproduces by continuous apomictic parthenogenesis. Genetic differentiation between localities was not significant, whereas a strong differentiation was observed between host plant families (0.175<FST<0.691). The great majority of aphids exhibited one of three predominant multilocus genotypes that were repeatedly and respectively associated to the three plant families, Cucurbitaceae, Solanaceae and Rutaceae, demonstrating host specialization in A. gossypii. These specialized genotypes were simultaneously found with other clones on Hibiscus, suggesting that this perennial host could act as a refuge plant between two vegetable crop seasons.


Insects ◽  
2020 ◽  
Vol 11 (4) ◽  
pp. 257 ◽  
Author(s):  
Steven Grodsky ◽  
Leslie Saul-Gershenz ◽  
Kara Moore-O’Leary ◽  
Rebecca Hernandez

Butterfly–host plant relationships can inform our understanding of ecological and trophic interactions that contribute to ecosystem function, resiliency, and services. The ecology of danaid–milkweed (Apocynaceae) host plant interactions has been studied in several biomes but is neglected in deserts. Our objective was to determine effects of plant traits, seasonality, and landscape-level host plant availability on selection of Mojave milkweed (Asclepias nyctaginifolia A. Gray) by ovipositing monarch butterflies (Danaus plexippus plexippus) and queen butterflies (Danaus gilippus thersippus) in the Californian Mojave Desert. We surveyed all known Mojave milkweed locations in the Ivanpah Valley, California (n = 419) during early, mid-, and late spring in 2017. For each survey, we counted monarch and queen butterfly eggs on each Mojave milkweed plant. We also measured canopy cover, height, volume, and reproductive stage of each Mojave milkweed plant. We counted a total of 276 queen butterfly eggs and zero monarch butterfly eggs on Mojave milkweed host plants. We determined that count of queen butterfly eggs significantly increased with increasing Mojave milkweed canopy cover. Additionally, count of queen butterfly eggs was: (1) greater on adult Mojave milkweed plants than on juvenile and seedling plants and greater on juvenile Mojave milkweed plants than on seedling plants; and (2) greater during early spring than mid-spring—we recorded no eggs during late spring. Based on aggregation indices, queen butterfly eggs occurred on Mojave milkweed plants in a nonrandom, clustered pattern throughout the Ivanpah Valley. We provide the first evidence of trophic interactions between queen butterflies and Mojave milkweed at multiple spatial scales in the Mojave Desert, suggesting that conservation and management practices for both species should be implemented concurrently. Given its role as an herbivore, pollinator and prey, the queen butterfly may serve as a model organism for understanding effects of anthropogenic disturbance (e.g., solar energy development) on “bottom-up” and trophic interactions among soils, plants and animals in desert ecosystems.


2021 ◽  
Author(s):  
Yu Okamura ◽  
Ai Sato ◽  
Lina Kawaguchi ◽  
Atsushi J. Nagano ◽  
Masashi Murakami ◽  
...  

Herbivorous insects have evolved counteradaptations to overcome the chemical defenses of their host plants. Several of these counteradaptations have been elucidated at the molecular level, in particular for insects specialized on cruciferous host plants. While the importance of these counteradaptations for host plant colonization is well established, little is known about their microevolutionary dynamics in the field. In this study, we examine patterns of host plant use and insect counteradaptation in three Pieris butterfly species across Japan. The larvae of these butterflies express nitrile-specifier protein (NSP) and its paralog major allergen (MA) in their gut to overcome the highly diversified glucosinolate-myrosinase defense system of their cruciferous host plants. Pieris napi and Pieris melete colonize wild Brassicaceae whereas Pieris rapae typically uses cultivated Brassica as a host, regardless of the local composition of wild crucifers. As expected, NSP and MA diversity was independent of the local composition of wild Brassicaceae in P. rapae. In contrast, NSP diversity correlated with local host plant diversity in both species that preferred wild Brassicaceae. P. melete and P. napi both revealed two distinct major NSP alleles, which shaped diversity among local populations, albeit with different evolutionary trajectories. In comparison, MA showed no indication for local adaptation. Altogether, MA appeared to be evolutionary more conserved than NSP, suggesting that both genes play different roles in diverting host plant chemical defense.


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