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Gustavo G L Costa,
Odalys G Cabrera,
Ricardo A Tiburcio,
Francisco J Medrano,
Marcelo F Carazzolle,
Daniela P T Thomazella,
Stephen C Schuster,
John E Carlson,
Mark J Guiltinan, Bryan A Bailey,
Piotr Mieczkowski,
Gonçalo A G Pereira,
Lyndel W Meinhardt
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ABSTRACT: In this study, we report the sequence of the mitochondrial (mt) genome of the Basidiomycete fungus Moniliophthora roreri, which is the etiologic agent of frosty pod rot of cacao (Theobroma cacao L.). We also compare it to the mtDNA from the closely-related species Moniliophthora perniciosa, which causes witches' broom disease of cacao. The 94 Kb mtDNA genome of M. roreri has a circular topology and codes for the typical 14 mt genes involved in oxidative phosphorylation. It also codes for both rRNA genes, a ribosomal protein subunit, 13 intronic open reading frames (ORFs), and a full complement of 27 tRNA genes. The conserved genes of M. roreri mtDNA are completely syntenic with homologous genes of the 109 Kb mtDNA of M. perniciosa. As in M. perniciosa, M. roreri mtDNA contains a high number of hypothetical ORFs (28), a remarkable feature that make Moniliophthoras the largest reservoir of hypothetical ORFs among sequenced fungal mtDNA. Additionally, the mt genome of M. roreri has three free invertron-like linear mt plasmids, one of which is very similar to that previously described as integrated into the main M. perniciosa mtDNA molecule. Moniliophthora roreri mtDNA also has a region of suspected plasmid origin containing 15 hypothetical ORFs distributed in both strands. One of these ORFs is similar to an ORF in the mtDNA gene encoding DNA polymerase in Pleurotus ostreatus. The comparison to M. perniciosa showed that the 15 Kb difference in mtDNA sizes is mainly attributed to a lower abundance of repetitive regions in M. roreri (5.8 Kb vs 20.7 Kb). The most notable differences between M. roreri and M. perniciosa mtDNA are attributed to repeats and regions of plasmid origin. These elements might have contributed to the rapid evolution of mtDNA. Since M. roreri is the second species of the genus Moniliophthora whose mtDNA genome has been sequenced, the data presented here contribute valuable information for understanding the evolution of fungal mt genomes among closely-related species.
Fungal Biology 05/2012; 116(5):551-62. · 1.43 Impact Factor
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ABSTRACT: Four taxa of the plant genus Erythroxylum; Erythroxylum coca var. coca (Ecc), Erythroxylum coca var. ipadu (Eci), Erythroxylum novogranatense var. novogranatense (Enn) and Erythroxylum novogranatense var. truxillense (Ent) are cultivated primarily for the illicit extraction and processing of cocaine. Despite their economic and medical importance,
the evolutionary history of these species remains unknown in a modern phylogenetic framework. The aims of this study were
to: (a) investigate the relationship among the cultivated and a select number of non-cultivated taxa, and (b) test Plowman’s
(Journal of Psychodelic Drugs 11:103–117, 1979b) linear progression hypothesis of the cultivated Erythroxylum taxa versus Johnson’s et al. (Annals of Botany 95:601–608, 2005) hypothesis that Ec and En are sister species. AFLP phylogeny was used to compare the relationships among 36 Erythroxylum species (133 accessions) spanning the geographic distribution of the genus. A Maximum Parsimony tree revealed both geographic
and taxonomic partitioning into clades representing species from Africa, Asia-Pacific and the New World (Tropical Americas).
Ec and En formed distinct clades, indicating they are sister species and a cluster of non-cultivated species were the most closely
related to the cultivated species. Multivariate ordination analysis was used to evaluate the relationship between cultivated
and non-cultivated Erythroxylum taxa from the Tropical Americas. Our results support the hypothesis that the cultivated species are more closely related
to each other than to any other species of Erythroxylum, but refute the hypothesis that Ent (and Enn) descended from Ecc. Instead our data suggest an independent, non-linear evolutionary relationship between Ec and En. Finally, the AFLP analyses identified significantly different genetic groups within Erythroxylum suggesting that the current intrageneric classification of this genus be revised.
KeywordsAFLP–Cocaine–
Erythroxylum
–Genetic diversity–Phylogenetics
Tropical Plant Biology 04/2012; 4(2):126-133.
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ABSTRACT: The aim of this study was to determine whether endophytic Bacillus cereus isolates from agronomic crops possessed genes for the nonhaemolytic enterotoxin (Nhe) and haemolysin BL (HBL) and, therefore, have the potential to cause diarrhoeal illness in humans.
PCR followed by sequencing confirmed the presence of enterotoxin genes nheA, nheB, nheC, hblA, hblC, hblD in endophytic B. cereus. All nhe genes were detected in 59% of endophytic B. cereus, while all hbl genes were detected in 44%. All six genes were detected in 41% of isolates. Enterotoxin genes were not detected in 15% of B. cereus isolates. Reverse transcriptase real-time PCR confirmed that endophytic B. cereus could express enterotoxin genes in pure culture.
This study showed that endophytic B. cereus isolates that possess genes for enterotoxin production are present in agronomic crops. Other endophytic B. cereus isolates lacked specific genes or lacked all nhe and hbl genes. Additionally, host, country of origin and tissue of origin had no impact on the enterotoxin genes detected.
Bacillus cereus with the potential of causing diarrhoeal illness in humans is a cosmopolitan endophytic inhabitant of plants, not incidental surface inhabitants or contaminants, as often suggested by previous research.
Letters in Applied Microbiology 03/2012; 54(5):468-74. · 1.62 Impact Factor
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ABSTRACT: Witches’ broom disease of Theobroma cacao L. is caused by the hemibiotrophic basidiomycete Moniliophthora perniciosa. Infection of flower cushions by M. perniciosa results in parthenocarpy. Healthy and parthenocarpic immature cacao pods were obtained from seven cacao clones. Microscopic observations of parthenocarpic pods from two clones confirmed that fruits lack viable seed. Septate mycelia colonized parthenocarpic pods, but were absent from healthy pods. Parthenocarpic pods had increased concentrations of leucine, methionine, serine, phenylalanine, and valine. Major transport metabolites sucrose and asparagine were decreased by 63 and 40 %, respectively, during parthenocarpy. M. perniciosa expressed sequence tags (ESTs) related to detoxification (MpSOD2 and MpCTA1) and nutrient acquisition (MpAS, MpAK, MpATG8, MpPLY, and MpPME) were induced in parthenocarpic pods. Most M. perniciosa ESTs related to plant hormone biosynthesis were repressed (MpGAox, MpCPS, MpDES, MpGGPPS, and MpCAO) in parthenocarpic pods. RT-qPCR analysis was conducted for 54 defense-related cacao ESTs and 93 hormone-related cacao ESTs. Specific cacao ESTs related to plant defense were induced (TcPR5, TcChi4, TcThau-ICS) while others were repressed (TcPR1, TcPR6, TcP12, and TcChiB). Cacao ESTs related to GA biosynthesis (TcGA20OX1B) were repressed in parthenocarpic pods. Cacao ESTs putatively related to maintaining cytokinin (TcCKX3 and TcCKX5) and IAA (TcGH3.17a, TcGH3.1, TcARF18) homeostasis were induced in parthenocarpic pods, suggesting an attempt to regulate cytokinin and auxin concentrations. In conclusion, M. perniciosa expresses specific sets of transcripts targeting nutrient acquisition and survival while altering the host physiology without causing significant necrosis resulting in parthenocarpy. Only a general host defense response is elicited.
Tree Genetics & Genomes 01/2012; 8(6):1261. · 2.34 Impact Factor
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ABSTRACT: a b s t r a c t Sixty-nine endospore-forming bacterial endophytes consisting of 15 different species from five genera were isolated from leaves, pods, branches, and flower cushions of Theobroma cacao as potential biological control agents. Sixteen isolates had in vitro chitinase production. In antagonism studies against cacao pathogens, 42% inhibited Moniliophthora roreri, 33% inhibited Moniliophthora perniciosa, and 49% inhib-ited Phytophthora capsici. Twenty-five percent of isolates inhibited the growth of both Moniliophthora spp., while 22% of isolates inhibited the growth of all three pathogens. Isolates that were chitinolytic and tested negative on Bacillus cereus agar were tested with in planta studies. All 14 isolates colonized the phyllosphere and internal leaf tissue when introduced with Silwet L-77, regardless of the tissue of origin of the isolate. Eight isolates significantly inhibited P. capsici lesion formation (p = 0.05) in detached leaf assays when compared to untreated control leaves. ARISA with bacilli specific primers amplified 21 OTUs in field grown cacao leaves, while eubacteria specific primers amplified 58 OTUs. ARISA analysis of treated leaves demonstrated that inundative application of a single bacterial species did not cause a long-term shift of native bacterial communities. This research illustrates the presence of endospore-forming bacterial endophytes in cacao trees, their potential as antagonists of cacao pathogens, and that cacao har-bors a range of bacterial endophytes. Published by Elsevier Inc.
04/2011;
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ABSTRACT: Endophytic Trichoderma isolates collected in tropical environments were evaluated for biocontrol activity against Phytophthora capsici in hot pepper (Capsicum annuum). Six isolates were tested for parasitic and antimicrobial activity against P. capsici and for endophytic and induced resistance capabilities in pepper. Isolates DIS 70a, DIS 219b, and DIS 376f were P. capsici parasites, while DIS 70a, DIS 259j, DIS 320c, and DIS 376f metabolites inhibited P. capsici. All six isolates colonized roots but were inefficient stem colonizers. DIS 259j, DIS 320c, and DIS 376f induced defense-related expressed sequence tags (EST) in 32-day-old peppers. DIS 70a, DIS 259j, and DIS 376f delayed disease development. Initial colonization of roots by DIS 259j or DIS 376f induced EST with potential to impact Trichoderma endophytic colonization and disease development, including multiple lipid transferase protein (LTP)-like family members. The timing and intensity of induction varied between isolates. Expression of CaLTP-N, encoding a LTP-like protein in pepper, in N. benthamiana leaves reduced disease development in response to P. nicotianae inoculation, suggesting LTP are functional components of resistance induced by Trichoderma species. Trichoderma isolates were endophytic on pepper roots in which, depending on the isolate, they delayed disease development by P. capsici and induced strong and divergent defense reactions.
Molecular Plant-Microbe Interactions 03/2011; 24(3):336-51. · 4.43 Impact Factor
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ABSTRACT: Theobroma cacao (cacao) is a tropical understory tree with sensitivity to drought. Cacao responds to drought by decreases in net photosynthesis,
PS II efficiency, stomatal conductance, water potential and changes in leaf florescence. Drought also alters cacao gene expression
as well as leaf glucose and free amino acid content. In recent years an incredible diversity of fungal endophytes has been
identified in association with cacao. These endophytes are being studied for the benefits they provide to cacao including
tolerance to biotic and abiotic stresses. During establishment of the endophytic association between cacao and fungal endophytes
both plant and fungal gene expression are altered. The endophytic Trichoderma hamatum isolate DIS 219b delays the onset of drought stress in cacao. This delay manifests itself through enhanced root growth, maintenance
of stomatal conductance, water potential, net photosynthesis, and PSII efficiency, changes in free amino acid concentrations,
and a delay in drought-induced changes in leaf gene expression. The cacao plant and DIS 219b adapt to each other and this
adaptation may contribute to the observed plant growth promotion and the delay in onset of drought stress. The increase in
root growth is thought to increase water uptake and availability, delaying the time point where the water supply becomes limiting
and drought stress occurs.
12/2010: pages 157-172;
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Ricardo Augusto Tiburcio,
Gustavo Gilson Lacerda Costa,
Marcelo Falsarella Carazzolle,
Jorge MC Mondego,
Stephen C. Schuster,
John E. Carlson,
Mark J. Guiltinan, Bryan A. Bailey,
Piotr Mieczkowski,
Lyndel W. Meinhardt,
Goncalo AG Pereira
Journal of Molecular Evolution 01/2010; · 2.27 Impact Factor
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ABSTRACT: Four biologically active cDNA clones were derived from the Alternanthera mosaic virus (AltMV; genus Potexvirus) isolate, AltMV-SP, which differ in symptoms in infected Nicotiana benthamiana plants. Two clones induced necrosis and plant death; a mixture of all four clones induced milder symptoms than AltMV-SP. Replication of all clones was enhanced by a minimum of fourfold at 15 degrees C. A mixture of clones 4-7 (severe) and 3-1 (mild) was indistinguishable from AltMV-SP, but the ratio of 4-7 to 3-1 differed at 25 and 15 degrees C. RNA copy numbers of mixed infections were always below those of 4-7 alone. Determinants of symptom severity were identified in both Pol and TGB1; the mildest (4-1) and most severe (3-7) clones differed at three residues in the 'core' Pol domain [R(1110)P, K(1121)R, R(1255)K] and one [S(1535)P] in the C-terminal Pol domain of RNA-dependent RNA polymerase, and one in TGB1 [P(88)L]. Pol [P(1110),R(1121),K(1255)]+TGB1(L(88))] always induced systemic necrosis at 15 degrees C. Gene exchanges of Pol and TGB1 each affected replication and symptom expression, with TGB1(P(88)) significantly reducing silencing suppression. The difference in silencing suppression between TGB1(P(88)) and TGB1(L(88)) was confirmed by an agroinfiltration assay. Further, co-expression of TGB1(P(88)) and TGB1(L(88)) resulted in interference in the suppression of silencing by TGB1(L(88)). Yeast two-hybrid analysis confirmed that TGB1(P(88)) and TGB1(L(88)) interact. These results identify a TGB1 residue that significantly affects replication and silencing suppression, but maintains full movement functions.
Journal of General Virology 01/2010; 91(Pt 1):277-87. · 3.36 Impact Factor
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Ricardo Augusto Tiburcio,
Gustavo Gilson Lacerda Costa,
Marcelo Falsarella Carazzolle,
Jorge Maurício Costa Mondego,
Stephen C Schuster,
John E Carlson,
Mark J Guiltinan, Bryan A Bailey,
Piotr Mieczkowski,
Lyndel W Meinhardt,
Gonçalo Amarante Guimarães Pereira
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ABSTRACT: Moniliophthora perniciosa and Moniliophthora roreri are phytopathogenic basidiomycete species that infect cacao causing two important diseases in this crop: "Witches' Broom" and "Frosty Pod Rot", respectively. The ability of species from this genus (Moniliophthora) to cause disease is exceptional in the family Marasmiaceae. Species in closely related genera including, Marasmius, Crinipellis, and Chaetocalathus, are mainly saprotrophs and are not known to cause disease. In this study, the possibility that this phytopathogenic lifestyle has been acquired by horizontal gene transfer (HGT) was investigated. A stringent genome comparison pipeline was used to identify potential genes that have been obtained by Moniliophthora through HGT. This search led to the identification of three genes: a metallo-dependent hydrolase (MDH), a mannitol phosphate dehydrogenase (MPDH), and a family of necrosis-inducing proteins (NEPs). Phylogenetic analysis of these genes suggests that Moniliophthora acquired NEPs from oomycetes, MDH from actinobacteria and MPDH from firmicutes. Based on the known gene functions and on previous studies of M. perniciosa infection and development, a correlation between gene acquisition and the evolution of the phytopathogenic genus Moniliophthora can be postulated.
Journal of Molecular Evolution 01/2010; 70(1):85-97. · 2.27 Impact Factor
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ABSTRACT: Trichoderma species are usually considered soil organisms that colonize plant roots, sometimes forming a symbiotic relationship. Recent studies demonstrate that Trichoderma species are also capable of colonizing the above ground tissues of Theobroma cacao (cacao) in what has been characterized as an endophytic relationship. Trichoderma species can be re-isolated from surface sterilized cacao stem tissue, including the bark and xylem, the apical meristem, and to a lesser degree from leaves. SEM analysis of cacao stems colonized by strains of four Trichoderma species (Trichoderma ovalisporum-DIS 70a, Trichoderma hamatum-DIS 219b, Trichoderma koningiopsis-DIS 172ai, or Trichoderma harzianum-DIS 219f) showed a preference for surface colonization of glandular trichomes versus non-glandular trichomes. The Trichoderma strains colonized the glandular trichome tips and formed swellings resembling appresoria. Hyphae were observed emerging from the glandular trichomes on surface sterilized stems from cacao seedlings that had been inoculated with each of the four Trichoderma strains. Fungal hyphae were observed under the microscope emerging from the trichomes as soon as 6h after their isolation from surface sterilized cacao seedling stems. Hyphae were also observed, in some cases, emerging from stalk cells opposite the trichome head. Repeated single trichome/hyphae isolations verified that the emerging hyphae were the Trichoderma strains with which the cacao seedlings had been inoculated. Strains of four Trichoderma species were able to enter glandular trichomes during the colonization of cacao stems where they survived surface sterilization and could be re-isolated. The penetration of cacao trichomes may provide the entry point for Trichoderma species into the cacao stem allowing systemic colonization of this tissue.
Mycological Research 09/2009; 113(Pt 12):1365-76. · 2.81 Impact Factor
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ABSTRACT: Theobroma cacao (cacao) is cultivated in tropical climates and is exposed to drought stress. The impact of the endophytic fungus Trichoderma hamatum isolate DIS 219b on cacao's response to drought was studied. Colonization by DIS 219b delayed drought-induced changes in stomatal conductance, net photosynthesis, and green fluorescence emissions. The altered expression of 19 expressed sequence tags (ESTs) (seven in leaves and 17 in roots with some overlap) by drought was detected using quantitative real-time reverse transcription PCR. Roots tended to respond earlier to drought than leaves, with the drought-induced changes in expression of seven ESTs being observed after 7 d of withholding water. Changes in gene expression in leaves were not observed until after 10 d of withholding water. DIS 219b colonization delayed the drought-altered expression of all seven ESTs responsive to drought in leaves by > or = 3 d, but had less influence on the expression pattern of the drought-responsive ESTs in roots. DIS 219b colonization had minimal direct influence on the expression of drought-responsive ESTs in 32-d-old seedlings. By contrast, DIS 219b colonization of 9-d-old seedlings altered expression of drought-responsive ESTs, sometimes in patterns opposite of that observed in response to drought. Drought induced an increase in the concentration of many amino acids in cacao leaves, while DIS 219b colonization caused a decrease in aspartic acid and glutamic acid concentrations and an increase in alanine and gamma-aminobutyric acid concentrations. With or without exposure to drought conditions, colonization by DIS 219b promoted seedling growth, the most consistent effects being an increase in root fresh weight, root dry weight, and root water content. Colonized seedlings were slower to wilt in response to drought as measured by a decrease in the leaf angle drop. The primary direct effect of DIS 219b colonization was promotion of root growth, regardless of water status, and an increase in water content which it is proposed caused a delay in many aspects of the drought response of cacao.
Journal of Experimental Botany 07/2009; 60(11):3279-95. · 5.36 Impact Factor
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ABSTRACT: A small assortment of microbial proteins have the ability to activate defense responses and induce necrosis in plant cells through cell signaling pathways. These proteins are of interest because of their potential use as bioherbicides and inducers of plant resistance in agriculture. A 24-kDa protein (Nep1) was purified from culture filtrates of Fusarium oxysporum, and the effects of this protein on weed leaves were investigated. This protein induced necrosis in detached leaves of Papaver somniferum, Lycopersicon esculentum, Malva neglecta, and Acroptilon repens when taken up through the petiole. The pattern and level of necrosis were dependent on the plant species. Treatment with Nep1 induced the production of ethylene in isolated leaves of various species, and the level of ethylene response was shown to be correlated to the concentration of the protein. Pretreating leaves of P. somniferum, L. esculentum, M. neglecta, and Cardaria draba with 100 µl L-1 ethylene enhanced the protein induction of ethylene biosynthesis in those leaves. Application of Nep1 (200 nM) as a spray to intact plants of Abutilon theophrasti, P. somniferum, Centaurea solstitialis, Centaurea maculosa, and Sonchus oleraceus resulted in extensive necrosis of leaves within 48 h. The results of this research are supplemental to our understanding of the role of specific polypeptides in plant/microbe interactions and demonstrates for the first time that a fungal protein can cause extensive necrosis when applied to weed species as a foliar spray. Nomenclature: Fusarium oxysporum Schlechtend:Fr. f. sp. erythroxyli; Malva ne-glecta Wallr., MALNE, common mallow; Cardaria draba L., CADDR, hoary cress; Papaver somniferum L., PAPSO, opium poppy; Acroptilon repens L. CENRE, Russian knapweed; Centaurea maculosa Lam., CENMA, spotted knapweed; Sonchus oleraceus L. SONOL, annual sowthistle; Abutilon theophrasti Medicus, ABUTH, velvetleaf; Centaurea solstitialis L. CENSO, yellow starthistle; Lycopersicon esculentum L. 'Bonnie Best,' tomato.
Weed Science 01/2009; · 1.73 Impact Factor
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ABSTRACT: The fungal protein Nep1, produced by Fusarium oxysporum f. sp. erythroxyli in liquid culture, caused extensive necrosis to Centaurea maculosa when water solutions of Nep1 (5 µg ml−1) and an organosilicone surfactant (1,1,1,3,5,5,5-heptamethyltrisiloxanyl propyl-methoxy-poly[ethylene oxide]) were applied as foliar sprays. Nep1 did not cause necrosis when applied with a nonionic surfactant or organosilicone surfactant plus unrefined corn oil. Plant age, protein concentration, organosilicone surfactant concentration, and the presence of a dew period influenced the amount of necrosis caused by Nep1. The addition of an 18-h dew period after treatment resulted in an increase of 10% or more in foliar necrosis at the 0.313 and 1.25 µg ml−1 (0.40 and 1.62 g ai ha−1) Nep1 concentrations. Increasing the spray volume from 129 ml m−2 (1,291.3 L ha−1) to 516 ml m−2 (5,165.2 L ha−1) more than doubled the amount of foliar necrosis caused by the 0.313 µg ml−1 (0.40 g ai ha−1 vs. 1.62 g ai ha−1) Nep1 concentration. A maximum necrosis rating of 95% was reached by 1.25 µg ml−1 Nep1 applied at 516 ml m−2 (6.46 g ai ha−1) followed by an 18-h dew period. Nep1 (6.46 g ai ha−1) remained active when coapplied to Centaurea maculosa with the herbicides 2,4-D or glyphosate (0.13 to 2.58 kg ai ha−1), causing foliar necrosis prior to the herbicides killing Centaurea maculosa. An increase in the organosilicone surfactant concentration from 1 to 2 ml ai L−1 was required to achieve levels of Nep1-induced necrosis on Centaurea maculosa acclimated to direct sun comparable to levels achieved on greenhouse-grown plants. Repeated application of Nep1 (6.48 g ai ha−1) 3 wk after an initial treatment (6.48 g ai ha−1) prevented the recovery of acclimated Centaurea maculosa. Greater damage was caused to acclimated Centaurea maculosa when Nep1 was applied near the middle of the day (80% necrosis at 10:00 a.m. and 85% necrosis at 2:00 p.m.) compared to early or late in the day (25% necrosis at 6:00 a.m. and 10% necrosis at 6:00 p.m.). Nomenclature: Glyphosate, monoisopropylamine salt of glyphosate; 2,4-D, sodium salt of 2,4-D; Centaurea maculosa Lam. CENMA, spotted knapweed; Fusarium oxysporum; Nep1.
Weed Science 01/2009; · 1.73 Impact Factor
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Jorge MC Mondego,
Marcelo F Carazzolle,
Gustavo GL Costa,
Eduardo F. Formighieri,
Lucas P Parizzi,
Johana Rincones,
Carolina Cotomacci,
Dirce M Carraro,
Anderson F Cunha,
Helaine Carrer, [......],
Karina P Gramacho,
Marilda S Gonçalves,
José P Moura Neto,
Aristóteles Góes-Neto,
Luciana V. Barbosa,
Mark J Guiltinan, Bryan A Bailey,
Lyndel W Meinhardt,
Julio CM Cascardo,
Gonçalo AG Pereira
BMC Genomics 11/2008; · 4.07 Impact Factor
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ABSTRACT: Moniliophthora perniciosa (=Crinipellis perniciosa) causes one of the three main fungal diseases of Theobroma cacao (cacao), the source of chocolate. This pathogen causes Witches' broom disease (WBD) and has brought about severe economic losses in all of the cacao-growing regions to which it has spread with yield reductions that range from 50 to 90%. Cacao production in South America reflects the severity of this pathogen, as the yields in most of the infected regions have not returned to pre-outbreak levels, even with the introduction of resistant varieties. In this review we give a brief historical account and summarize the current state of knowledge focusing on developments in the areas of systematics, fungal physiology, biochemistry, genomics and gene expression in an attempt to highlight this disease. Moniliophthora perniciosa is a hemibiotrophic fungus with two distinct growth phases. The ability to culture a biotrophic-like phase in vitro along with new findings derived from the nearly complete genome and expression studies clearly show that these different fungal growth phases function under distinct metabolic parameters. These new findings have greatly improved our understanding of this fungal/host interaction and we may be at the crossroads of understanding how hemibiotrophic fungal plant pathogens cause disease in other crops. HISTORICAL SUMMARY OF WBD: The first WDB symptoms appear to have been described in the diaries of Alexandre Rodrigues Ferreira (described as lagartão; meaning big lizard) from his observations of cacao trees in 1785 and 1787 in Amazonia, which is consistent with the generally accepted idea that M. perniciosa, like its main host T. cacao, evolved in this region. The disease subsequently arrived in Surinam in 1895. WBD moved rapidly, spreading to Guyana in 1906, Ecuador in 1918, Trinidad in 1928, Colombia in 1929 and Grenada in 1948. In each case, cacao production was catastrophically affected with yield reductions of 50-90%. After the arrival of M. perniciosa in Bahia in 1989, Brazil went from being the world's 3rd largest producer of cacao (347 000 tonnes in 1988-1990; c. 15% of the total world production at that time) to a net importer (141 000 tonnes in 1998-2000). Fortunately for chocolate lovers, other regions of the world such as West Africa and South East Asia have not yet been affected by this disease and have expanded production to meet growing world demand (predicted to reach 3 700 000 tonnes by 2010). CLASSIFICATION: Moniliophthora perniciosa (Stahel) Aime & Phillips-Mora: super-kingdom Eukaryota; kingdom Fungi; phylum Basidiomycota; subphylum Agaricomycotina; class Agaricomycetes; subclass Agaricomycetidae; order Agaricales; family Marasmiaceae; genus Moniliophthora. USEFUL WEBSITES: http://www.lge.ibi.unicamp.br/vassoura/, http://nt.ars-grin.gov/taxadescriptions/keys/TrichodermaIndex.cfm, http://www.worldcocoafoundation.org/info-center/research-updates.asp, http://www.ars.usda.gov/ba/psi/spcl.
Molecular Plant Pathology 09/2008; 9(5):577-88. · 3.90 Impact Factor
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ABSTRACT: Drought can negatively impact pod production despite the fact that cacao production usually occurs in tropical areas having high rainfall. Polyamines (PAs) have been associated with the response of plants to drought in addition to their roles in responses to many other stresses. The constitutive and drought inducible expression patterns of genes encoding enzymes involved in PA biosynthesis were determined: an ornithine decarboxylase (TcODC), an arginine decarboxylase (TcADC), an S-adenosylmethionine decarboxylase (TcSAMDC), a spermidine synthase (TcSPDS), and a spermine synthase (TcSPMS). Expression analysis using quantitative real-time reverse transcription-PCR (QPCR) results showed that the PA biosynthesis genes were expressed in all plant tissues examined. Constitutive expression of PA biosynthesis genes was generally highest in mature leaves and open flowers. Expression of TcODC, TcADC, and TcSAMDC was induced with the onset of drought and correlated with changes in stomatal conductance, photosynthesis, photosystem II efficiency, leaf water potential and altered emission of blue-green fluorescence from cacao leaves. Induction of TcSAMDC in leaves was most closely correlated with changes in water potential. The earliest measured responses to drought were enhanced expression of TcADC and TcSAMDC in roots along with decreases in stomatal conductance, photosynthesis, and photosystem II efficiency. Elevated levels of putrescine, spermidine, and spermine were detected in cacao leaves 13days after the onset of drought. Expression of all five PA associated transcripts was enhanced (1.5-3-fold) in response to treatment with abscisic acid. TcODC and TcADC, were also responsive to mechanical wounding, infection by Phytophthora megakarya (a causal agent of black pod disease in cacao), the necrosis- and ethylene-inducing protein (Nep1) of Fusarium oxysporum, and flower abscission. TcSAMDC expression was responsive to all stresses except flower abscission. TcODC, although constitutively expressed at much lower levels than TcADC, TcSAMDC, TcSPDS, and TcSPMS, was highly inducible by the fungal protein Nep1 (135-fold) and the cacao pathogen Phytophthora megakarya (671-fold). The full length cDNA for ODC was cloned and characterized. Among the genes studied, TcODC, TcADC, and TcSAMDC were most sensitive to induction by drought in addition to other abiotic and biotic stresses. TcODC, TcADC, and TcSAMDC may share signal transduction pathways and/or the stress induced signal induction pathways may converge at these three genes leading to similar although not identical patterns of expression. It is possible altering PA levels in cacao will result in enhanced tolerance to multiple stresses including drought and disease as has been demonstrated in other crops.
Plant Physiology and Biochemistry 03/2008; 46(2):174-88. · 2.84 Impact Factor
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ABSTRACT: In this study, we investigated protein and genetic profiles of Kunitz trypsin inhibitors (KTIs) in seeds of 16 different soybean genotypes that included four groups consisting of wild soybean (Glycine soja), the cultivated soybean (G. max) ancestors of modern N. American soybean cultivars (old), modern N. American soybean (elite), and Asian cultivated soybean landraces that were the immediate results of domestication from the wild soybean. Proteins were well separated by two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) and stained protein cut from a 2D-PAGE indicated that KTI exists as multiple isoforms (spots) in soybean. Protein spots of KTI were identified and characterized using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF-MS). Although overall distribution patterns of the KTI protein spots appeared similar, the number and intensity of the protein spots between wild and cultivated genotypes varied. Three KTI peptides were identified in three of the wild genotypes, PI 393551, PI 407027 and PI 407282, in which KTI3 peptide showed highest intensity. The remaining wild genotype, PI 366120, showed four protein spots. In contrast, the ancestors, modern and Asian landrace genotypes showed only two protein spots corresponding to KTI. On the basis of DNA blot analysis, there is one copy of the KTI3 gene in all 16 genotypes. Polymorphism was detected in one of the wild genotypes (PI 366120) both in proteomic and genomic analyses. Our data suggest that the major variation of protein profiles were between wild and cultivated soybean genotypes rather than among genotypes in the same group. Genetic variation of KTI1, KTI2 and KTI3-related genes were detected within and between groups.
Journal of Plant Physiology 07/2007; 164(6):756-63. · 2.79 Impact Factor
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ABSTRACT: Treatment of Arabidopsis (Arabidopsis thaliana) with a necrosis- and ethylene-inducing peptide (Nep1) from Fusarium oxysporum inhibited both root and cotyledon growth and triggered cell death, thereby generating necrotic spots. Nep1-like proteins are produced by divergent microbes, many of which are plant pathogens. Nep1 in the plant was localized to the cell wall and cytosol based on immunolocalization results. The ratio of chlorophyll a fluorescence (F685 nm/F730 nm) significantly decreased after 75-min treatment with Nep1 in comparison to the control. This suggested that a short-term compensation of photosynthesis occurred in response to localized damage to cells. The concentrations of most water-soluble metabolites analyzed were reduced in Arabidopsis seedlings after 6 h of Nep1 treatment, indicating that the integrity of cellular membranes had failed. Microarray results showed that short-term treatment with Nep1 altered expression of numerous genes encoding proteins putatively localized to organelles, especially the chloroplast and mitochondria. Short-term treatment with Nep1 induced multiple classes of genes involved in reactive oxygen species production, signal transduction, ethylene biosynthesis, membrane modification, apoptosis, and stress. Quantitative PCR was used to confirm the induction of genes localized in the chloroplast, mitochondria, and plasma membrane, and genes responsive to calcium/calmodulin complexes, ethylene, jasmonate, ethylene biosynthesis, WRKY, and cell death. The majority of Nep1-induced genes has been associated with general stress responses but has not been critically linked to resistance to plant disease. These results are consistent with Nep1 facilitating cell death as a component of diseases caused by necrotrophic plant pathogens.
Plant physiology 08/2006; 141(3):1056-67. · 6.53 Impact Factor
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ABSTRACT: Phvytophthora megakarya is a devastating oomycete pathogen that causes black pod disease in cacao. Phytophthora species produce a protein that has a similar sequence to the necrosis and ethylene inducing protein (Nep1) of Fusarium oxysporum. Multiple copies of NEP1 orthologs (PmegNEP) have been identified in P. megakarya and four other Phytophthora species (P. citrophthora, P. capsici, P. palmivora, and P. sojae). Genome database searches confirmed the existence of multiple copies of NEP1 orthologs in P. sojae and P. ramorum. In this study, nine different PmegNEP orthologs from P. megakarya strain Mk-1 were identified and analyzed. Of these nine orthologs, six were expressed in mycelium and in P. megakarya zoospore-infected cacao leaf tissue. The remaining two clones are either regulated differently, or are nonfunctional genes. Sequence analysis revealed that six PmegNEP orthologs were organized in two clusters of three orthologs each in the P. megakarya genome. Evidence is presented for the instability in the P. megakarya genome resulting from duplications, inversions, and fused genes resulting in multiple NEP1 orthologs. Traits characteristic of the Phytophthora genome, such as the clustering of NEP1 orthologs, the lack of CATT and TATA boxes, the lack of introns, and the short distance between ORFs were also observed.
Mycological Research 01/2006; 109(Pt 12):1373-85. · 2.81 Impact Factor
