The study characterized Meloidogyne species infecting banana roots across plantations in Martinique, Guadeloupe, and French Guiana. A total of 96 nematode isolates were collected and identified using esterase and malate dehydrogenase enzyme phenotypes, as well as perineal patterns. The major species found were M. arenaria at 61.9% of isolates and M. incognita at 34.3%. Intraspecific variability was detected within M. arenaria and M. incognita using the enzyme analysis. RAPD markers also revealed genetic variation within and between species, with M. arenaria showing 61.6% polymorphic variation within isolates. This high intraspecific diversity in M. aren
Diversity of hymenopteran parasitoids (Hymenoptera: Chalcididae) associated w...arboreo.net
This research evaluated the diversity of hymenopteran
parasitoids (Hymenoptera: Chalcididae) at different
reforestation sites of Tectona grandis. Insects were collected with Malaise traps from October 2009 to September 2010.
One collected a total of 414 Chalcididae specimens
distributed in 3 genera and 16 species. Brachymeria and
Conura were the most representative genera with 14 species.
The site bordered by pasture vegetation presented a higher
number of collected specimens when compared to the other sites. Brachymeria pandora and Ceyxia ventrispinosa
occurred as super dominant, super abundant, super frequent and constant species.
Floating Row Cover & Transparent Mulch to Reduce Insect Populations, Virus Diseases & Increase Yield in Cantaloupe; Gardening Guidebook for Florida www.scribd.com/doc/239851313 ~ Florida Master Gardeners, Florida State University, For more information, Please see Organic Edible Schoolyards & Gardening with Children www.scribd.com/doc/239851214 - Double Food Production from your School Garden with Organic Tech www.scribd.com/doc/239851079 - Free School Gardening Art Posters www.scribd.com/doc/239851159 - Increase Food Production with Companion Planting in your School Garden www.scribd.com/doc/239851159 - Healthy Foods Dramatically Improves Student Academic Success www.scribd.com/doc/239851348 - City Chickens for your Organic School Garden www.scribd.com/doc/239850440 - Huerto Ecológico, Tecnologías Sostenibles, Agricultura Organica www.scribd.com/doc/239850233 - Simple Square Foot Gardening for Schools, Teacher Guide www.scribd.com/doc/23985111 ~
Identification and pathogenicity of fusarium and phomopsis foliar diseases of...Premier Publishers
Research on foliage disease of Jatropha curcas was conducted in Sokoto, Kebbi and Zamfara States of Nigeria to determine the occurrence, incidence and severity of the diseases. Fusarium and Phomopsis species were the fungal pathogens found to be responsible for the disease on J. curcas in the study area. A spore count of the isolates was made and used as inocula in the pathogenicity trial in glasshouse of the department to prove Kochs’ postulate. Results from the farmers’ field revealed that, highest incidence (81.00%) and severity (53.33%) of Phomopsis leaf blight was recorded in Tsaki of Sokoto State, while Janbaki in Kebbi State had the highest incidence (75.33%) and severity (60.00%) of Fusarium leaf blight. The surveys conducted showed that, J. curcas planted in lowland areas tend to be more prone to the fungal leaf blight particularly those close to water source. In the pathogenicity trial, results indicated that, there was no significant difference in the methods of inoculation and number of days after inoculation with respect to incidence and severity of leaf blight. It is recommended that fungicides that can be used for the management of fungal leaf blight of J. curcas should be identified.
Characterization of Arsenic contaminated Rice (Oryza Sativa L.) through RAPD ...IOSR Journals
Rice being cultivated under anaerobic condition is vulnerable to arsenite, a mobile arsenic speciation Both arsenate and arsenite are highly toxic to human body. Experiments were conducted in Nonaghata (Nadia) and De-ganga (North 24 parganas) with 40 genotypes of rice in Boro season. A few varieties have been identified with low arsenic. Simultaneously some of them were characterized at molecular level by RAPD technique. It is worth to be seen how the low grain and high grain arsenic varieties behave in RAPD fingerprinting. Fourteen out of the 18 decamer random primers showed amplification of genomic DNA in all individuals. A total of 59 fragments were scored, of these fragments, 10 (16.94%) were common to all accessions, 49 (83.06%) were polymorphic and shared by at least eight accessions. It is interesting to mention that the genotypes Azucena and Lemont have already been identified as low grain arsenic genotypes and occupied a distinct different cluster for all primers at the molecular level also. More research is needed in arsenic research in crop plants in different agro climatic situation to have a meaningful and stable conclusion so that the farmers and also the people of West Bengal do not suffer from arsenic hazards at least from the consumption of rice.
Diversity of hymenopteran parasitoids (Hymenoptera: Chalcididae) associated w...arboreo.net
This research evaluated the diversity of hymenopteran
parasitoids (Hymenoptera: Chalcididae) at different
reforestation sites of Tectona grandis. Insects were collected with Malaise traps from October 2009 to September 2010.
One collected a total of 414 Chalcididae specimens
distributed in 3 genera and 16 species. Brachymeria and
Conura were the most representative genera with 14 species.
The site bordered by pasture vegetation presented a higher
number of collected specimens when compared to the other sites. Brachymeria pandora and Ceyxia ventrispinosa
occurred as super dominant, super abundant, super frequent and constant species.
Floating Row Cover & Transparent Mulch to Reduce Insect Populations, Virus Diseases & Increase Yield in Cantaloupe; Gardening Guidebook for Florida www.scribd.com/doc/239851313 ~ Florida Master Gardeners, Florida State University, For more information, Please see Organic Edible Schoolyards & Gardening with Children www.scribd.com/doc/239851214 - Double Food Production from your School Garden with Organic Tech www.scribd.com/doc/239851079 - Free School Gardening Art Posters www.scribd.com/doc/239851159 - Increase Food Production with Companion Planting in your School Garden www.scribd.com/doc/239851159 - Healthy Foods Dramatically Improves Student Academic Success www.scribd.com/doc/239851348 - City Chickens for your Organic School Garden www.scribd.com/doc/239850440 - Huerto Ecológico, Tecnologías Sostenibles, Agricultura Organica www.scribd.com/doc/239850233 - Simple Square Foot Gardening for Schools, Teacher Guide www.scribd.com/doc/23985111 ~
Identification and pathogenicity of fusarium and phomopsis foliar diseases of...Premier Publishers
Research on foliage disease of Jatropha curcas was conducted in Sokoto, Kebbi and Zamfara States of Nigeria to determine the occurrence, incidence and severity of the diseases. Fusarium and Phomopsis species were the fungal pathogens found to be responsible for the disease on J. curcas in the study area. A spore count of the isolates was made and used as inocula in the pathogenicity trial in glasshouse of the department to prove Kochs’ postulate. Results from the farmers’ field revealed that, highest incidence (81.00%) and severity (53.33%) of Phomopsis leaf blight was recorded in Tsaki of Sokoto State, while Janbaki in Kebbi State had the highest incidence (75.33%) and severity (60.00%) of Fusarium leaf blight. The surveys conducted showed that, J. curcas planted in lowland areas tend to be more prone to the fungal leaf blight particularly those close to water source. In the pathogenicity trial, results indicated that, there was no significant difference in the methods of inoculation and number of days after inoculation with respect to incidence and severity of leaf blight. It is recommended that fungicides that can be used for the management of fungal leaf blight of J. curcas should be identified.
Characterization of Arsenic contaminated Rice (Oryza Sativa L.) through RAPD ...IOSR Journals
Rice being cultivated under anaerobic condition is vulnerable to arsenite, a mobile arsenic speciation Both arsenate and arsenite are highly toxic to human body. Experiments were conducted in Nonaghata (Nadia) and De-ganga (North 24 parganas) with 40 genotypes of rice in Boro season. A few varieties have been identified with low arsenic. Simultaneously some of them were characterized at molecular level by RAPD technique. It is worth to be seen how the low grain and high grain arsenic varieties behave in RAPD fingerprinting. Fourteen out of the 18 decamer random primers showed amplification of genomic DNA in all individuals. A total of 59 fragments were scored, of these fragments, 10 (16.94%) were common to all accessions, 49 (83.06%) were polymorphic and shared by at least eight accessions. It is interesting to mention that the genotypes Azucena and Lemont have already been identified as low grain arsenic genotypes and occupied a distinct different cluster for all primers at the molecular level also. More research is needed in arsenic research in crop plants in different agro climatic situation to have a meaningful and stable conclusion so that the farmers and also the people of West Bengal do not suffer from arsenic hazards at least from the consumption of rice.
Genetic variability and phylogenetic relationships studies of Aegilops L. usi...Innspub Net
Studying of genetic relationships among Aegilops L. species is very important for broadening the cultivated wheat genepool, and monitoring genetic erosion, because the genus Aegilops includes the wild relatives of cultivated wheat which contain numerous unique alleles that are absent in modern wheat cultivars and it can contribute to broaden the genetic base of wheat and improve yield, quality and resistance to biotic and abiotic stresses of wheat. The use of molecular markers, revealing polymorphism at the DNA level, has been playing an increasing part in plant biotechnology and their genetics studies. There are different types of markers, morphological, biochemical and DNA based molecular markers. These DNA-based markers based on PCR (RAPD, AFLP, SSR, ISSR, IRAP), amongst others, the microsatellite DNA marker has been the most widely used, due to its easy use by simple PCR, followed by a denaturing gel electrophoresis for allele size determination, and to the high degree of information provided by its large number of alleles per locus. Day by day development of such new and specific types of markers makes their importance in understanding the genomic variability and the diversity between the same as well as different species of the plants. In this review, we will discuss about genetic variability and phylogenetic relationships studies of Aegilops L. using some molecular markers, with theirs Advantages, and disadvantages.
We evaluated the oviposition preference and damage capacity of Spodoptera frugiperda on the different phenological stages of corn. Tests were performed at the Assis Chateaubriand Agricultural School (07º10'15" S, 35º51'13" W, altitude 634 meters), municipality of Lagoa Seca, Paraíba State, Brazil, in two areas of 500 m2, with CMS maize hybrid strain and maize intercropped with bean with the spacing of 0.80 x 0.40 m. Eggs and caterpillars were collected weekly on 50 plants randomly sampled in five spots. Height and number of leaves per plant, and damage from caterpillars of S. frugiperda were recorded using the scale, the rangers were., 0) no damage, 1) leaf scraped, 2) leaf pierced, 3) leaf torn, 4) damage in cartridge, 5) cartridge destroyed. The average number of clutches did not differ significantly among the three phenological stages of the culture, but average clutch size (number of eggs) was significantly smaller for the stage of 4-6 leaves. However, there was a significant interaction with respect to the number of clutches between position in the plant (lower, middle, and upper) and phenological stage, and between leaf surface and phenological stages. There were significant differences among tillage systems for corn in monoculture and corn intercropped with bean.
Assessment of Genetic Diversity in 13 Local Banana (Musa Spp.) Cultivars Usin...paperpublications3
Abstract: A Study was conducted to investigate the genetic variability among 13 local banana cultivars using 3 SSR primers of Mb1-69, Mb1-113 and Mb1-134. All the primer pairs amplified a total of 29 different marker bands with an average of 9.6 bands per primer. Among the 29 bands only 4 bands were monomorphic and the rest 25 bands were polymorphic. The sizes of the amplified DNA bands in 13 local banana cultivars varied from 200 bp to 600 bp. The primer Mb1-113 amplified the highest (14) number of DNA bands and the primer Mb1-69 amplified the lowest (7) number of DNA bands whilst primer Mb1-134 amplified 8 DNA bands. The values of pair-wise genetic distances ranged from 1.00 to 9.00 indicating the presence of wide genetic diversity. The dendogram constructed based on phylogenetic relationship analysis revealed that the highest genetic diversity (9.00) found between the cultivars champa and jawayta and also the cultivars champa and jahazy whilst the lowest (1.00) between the cultivars doubled haploid and kathaly, doubled haploid and sorishafruity, doubled haploid and amritsagor and doubled haploid and ganasundory. The UPGMA dendogram has segregated the 13 local banana cultivars into two major clusters. Agnishwar and champa formed in cluster 1 and the rest of the cultivars like sobri jesore, sobri, anazy, kathaly, jawayta, sorishafruity, amritsagor, jahazy, bangle, ganasundory and doubled haploid have constituted the cluster 2.
S M Masiul Azam, Md Shahidul Islam, Parvin Shahanaz, Md Shafiqur Rahman and Sarder Md Shahriar Alam. “Molecular Characterization of Brassica Cultivars through RAPD Markers” United International Journal for Research & Technology (UIJRT) 1.3 (2019): 41-45.
Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.
Assessment of Genetic Diversity in Sesame (Sesamum indicum L.) Genotypes at B...Premier Publishers
Field experiment was conducted to assess the extent of genetic diversity in sesame (Sesamum indicum L.) genotypes to identify superior genotypes for further improvement program. A total of forty-nine sesame genotypes were evaluated at Bako and Uke during 2018 cropping season. Data were recorded and analyzed by SAS software. The combined analysis showed significant differences among the genotypes for all traits. Cluster analysis grouped 49 sesame genotypes into four clusters. The highest inter-cluster distance occurred between clusters three and four while the lowest was between clusters one and two. Principal components analysis showed that about 76.1% of the total variations among sesame genotypes were contributed by the first four PCs with eigen values greater than unity. Estimation of phenotypic diversity based on qualitative traits showed seed color and flower color were the highest divergent traits followed by stem color and leaf color. Generally, the result of the study showed existence of significant genetic variability among tested genotypes. Therefore, simple selection of promising genotypes and crossing of highly divergent group to produce best heterotic offspring could be recommended from the present study.
Genetic variability and phylogenetic relationships studies of Aegilops L. usi...Innspub Net
Studying of genetic relationships among Aegilops L. species is very important for broadening the cultivated wheat genepool, and monitoring genetic erosion, because the genus Aegilops includes the wild relatives of cultivated wheat which contain numerous unique alleles that are absent in modern wheat cultivars and it can contribute to broaden the genetic base of wheat and improve yield, quality and resistance to biotic and abiotic stresses of wheat. The use of molecular markers, revealing polymorphism at the DNA level, has been playing an increasing part in plant biotechnology and their genetics studies. There are different types of markers, morphological, biochemical and DNA based molecular markers. These DNA-based markers based on PCR (RAPD, AFLP, SSR, ISSR, IRAP), amongst others, the microsatellite DNA marker has been the most widely used, due to its easy use by simple PCR, followed by a denaturing gel electrophoresis for allele size determination, and to the high degree of information provided by its large number of alleles per locus. Day by day development of such new and specific types of markers makes their importance in understanding the genomic variability and the diversity between the same as well as different species of the plants. In this review, we will discuss about genetic variability and phylogenetic relationships studies of Aegilops L. using some molecular markers, with theirs Advantages, and disadvantages.
We evaluated the oviposition preference and damage capacity of Spodoptera frugiperda on the different phenological stages of corn. Tests were performed at the Assis Chateaubriand Agricultural School (07º10'15" S, 35º51'13" W, altitude 634 meters), municipality of Lagoa Seca, Paraíba State, Brazil, in two areas of 500 m2, with CMS maize hybrid strain and maize intercropped with bean with the spacing of 0.80 x 0.40 m. Eggs and caterpillars were collected weekly on 50 plants randomly sampled in five spots. Height and number of leaves per plant, and damage from caterpillars of S. frugiperda were recorded using the scale, the rangers were., 0) no damage, 1) leaf scraped, 2) leaf pierced, 3) leaf torn, 4) damage in cartridge, 5) cartridge destroyed. The average number of clutches did not differ significantly among the three phenological stages of the culture, but average clutch size (number of eggs) was significantly smaller for the stage of 4-6 leaves. However, there was a significant interaction with respect to the number of clutches between position in the plant (lower, middle, and upper) and phenological stage, and between leaf surface and phenological stages. There were significant differences among tillage systems for corn in monoculture and corn intercropped with bean.
Assessment of Genetic Diversity in 13 Local Banana (Musa Spp.) Cultivars Usin...paperpublications3
Abstract: A Study was conducted to investigate the genetic variability among 13 local banana cultivars using 3 SSR primers of Mb1-69, Mb1-113 and Mb1-134. All the primer pairs amplified a total of 29 different marker bands with an average of 9.6 bands per primer. Among the 29 bands only 4 bands were monomorphic and the rest 25 bands were polymorphic. The sizes of the amplified DNA bands in 13 local banana cultivars varied from 200 bp to 600 bp. The primer Mb1-113 amplified the highest (14) number of DNA bands and the primer Mb1-69 amplified the lowest (7) number of DNA bands whilst primer Mb1-134 amplified 8 DNA bands. The values of pair-wise genetic distances ranged from 1.00 to 9.00 indicating the presence of wide genetic diversity. The dendogram constructed based on phylogenetic relationship analysis revealed that the highest genetic diversity (9.00) found between the cultivars champa and jawayta and also the cultivars champa and jahazy whilst the lowest (1.00) between the cultivars doubled haploid and kathaly, doubled haploid and sorishafruity, doubled haploid and amritsagor and doubled haploid and ganasundory. The UPGMA dendogram has segregated the 13 local banana cultivars into two major clusters. Agnishwar and champa formed in cluster 1 and the rest of the cultivars like sobri jesore, sobri, anazy, kathaly, jawayta, sorishafruity, amritsagor, jahazy, bangle, ganasundory and doubled haploid have constituted the cluster 2.
S M Masiul Azam, Md Shahidul Islam, Parvin Shahanaz, Md Shafiqur Rahman and Sarder Md Shahriar Alam. “Molecular Characterization of Brassica Cultivars through RAPD Markers” United International Journal for Research & Technology (UIJRT) 1.3 (2019): 41-45.
Dioscorea rotundata is a staple food crop for millions of people in the tropical and subtropical regions. In vitro germplasm conservation is a very useful tool in yam improvement strategies but very little is known about the genetic integrity and stability of in-vitro conserved yam plants. In this study, 42 accessions from in vitro and field populations were genotyped using 11 microsatellite markers and 23 morphological descriptors to assess variability within and between accessions. Out of the 23 morphological variables used, 13 were identified as most discriminate and were used to cluster the accessions into 4 clusters using the unweighted pair group arithmetic mean average (UPGMA). Accession maintained in field as well as in in-vitro showed high genetic similarity (R2 = 0.91, p-value: 1e-04). Out of the 42 accessions analyzed, nine accessions maintained in the field and in-vitro displayed different genetic profiles. This study provided basic information on the possible somaclonal variation of yam accessions maintained through in-vitro. Further study with advanced tools such as next-generation sequencing is required to elucidate the nature of the observed variation within clones.
Assessment of Genetic Diversity in Sesame (Sesamum indicum L.) Genotypes at B...Premier Publishers
Field experiment was conducted to assess the extent of genetic diversity in sesame (Sesamum indicum L.) genotypes to identify superior genotypes for further improvement program. A total of forty-nine sesame genotypes were evaluated at Bako and Uke during 2018 cropping season. Data were recorded and analyzed by SAS software. The combined analysis showed significant differences among the genotypes for all traits. Cluster analysis grouped 49 sesame genotypes into four clusters. The highest inter-cluster distance occurred between clusters three and four while the lowest was between clusters one and two. Principal components analysis showed that about 76.1% of the total variations among sesame genotypes were contributed by the first four PCs with eigen values greater than unity. Estimation of phenotypic diversity based on qualitative traits showed seed color and flower color were the highest divergent traits followed by stem color and leaf color. Generally, the result of the study showed existence of significant genetic variability among tested genotypes. Therefore, simple selection of promising genotypes and crossing of highly divergent group to produce best heterotic offspring could be recommended from the present study.
This study was carried out on the mycoflora associated with seeds of different citrus species. Citrus seed material was collected from districts of Punjab, i.e. Multan, Sargodha and Khanpur. Standard methods were applied for the isolation and identification of fungi. A total of 11 fungi including Aspergillus fumigatus, Aspergillus flavus, Dreschslera tetramera, Alternaria alternata, Curvularia lunata, Macrophomina phaseolina, Aspergillus niger, Fusarium solani, Fusarium moniliforme, Rhizopus and Penicillium spp were isolated from the seeds of citrus. For control of isolated seed-born fungi, 3 recommended fungicides such as Ridomil Gold, Bavistin, Score and two chemical Salicylic acid and Boric acid, were used at 20, 30, 40 mg/10 mL and 5, 6, 7 μL/10 mL, respectively and chemical with 20, 30, 40 mg/10 mL. All these fungicide and chemicals significantly reuced with population of all fungi present in naturally infected seed samples. Ridomil Gold and Salicylic acid were found to be the best for the control of se d-born fungi of citrus seed at 40 mg/10 mL. The isolation and identification of different mycotoxins is essential to study health status of the citrus consumers and to safeguard the standards of WTO.
Genome Sequencing in Finger Millet
Genome size estimation
SOLiD Sequencing Technology
Illumina Sequencing Technology
Gene prediction and functional annotation of genes
Mining of plant transcription factors and other genes
SDS-PAGE electrophoretic analysis of young leaves protein was used to asses the genetic relatedness
among 54 specimens belonging to 6 common cultivars of Phoenix dactylifera L. (Aglany, Amry, Haiany,
Bent Aisha, Samany, and Zaghloul), that were collected from 7 different locations in Ismailia region. A
total of 10 different protein bands were collectively detected in the gels of the 54 studied specimens. The
resulted protein profile comprised one monomorphic band. The maximum number of bands observed was
10 bands found in one specimen of Zaghloul cultivar. While the minimum number of bands observed
was three bands found in one specimen of Aglany cultivar and two specimens of Bent Aisha cultivar. The
data of the allele frequencies of the six studied date palm cultivars revealed that all the alleles in Haiany
cultivar were monomorphic, except in three loci with the lowest percentage of polymorphic loci (30%).
While Bent Aisha and Aglany cultivars have the highest polymorphism (70% and 80%, respectively). In
addition, it was found that the ratio of gene diversity/locus varied greatly within the specimens of the six
studied date palm cultivars. Agglomerative cluster analysis, based on the genetic distances of the studied
54 specimens, revealed the variations and relatedness among the six collected cultivars.
Morphological diversity, pathogenicity and biofungicides efficacity on Cercos...Open Access Research Paper
Fungal diseases are one of major constraints on groundnut production in Burkina Faso. Among these diseases, early leaf spot caused by Cercospora arachidicola (Hori.) is one of the most important economic diseases of groundnut. Aim of contributing to search effective control methods against this disease, we undertook the present study, which consisted in (i) study morphological diversity of different Cercospora arachidicola isolates (ii) study the level of pathogenicity of Cercospora arachidicola strains identified (iii) evaluate the efficacy of some bio-fungicides on the strains identified. The study was carried out in 14 villages in the Hauts Bassins and Boucle du Mouhoun regions of Burkina Faso with regard to prospecting and sample collection, and at the INERA Bobo Dioulasso plant pathology laboratory for isolation, identification, pathogenicity and biofungicide efficacy testing. A total of five strains of Cercospora arachidicola were identified. The pathogenicity test was used to classify the five strains according to their virulence. In decreasing order of virulence, the strains were Fara, Darsa, Logo, Santi and Kod. In vitro evaluation of biofungicide efficacy shows that PLANSAIN biofungicide provides better control of Cercospora arachidicola strains. Trichoderma hazanium, the active ingredient in PLANSAIN, inhibits the radial growth of Cercospora arachidicola strains to a greater extent.
Identification and marketing of Marantaceae in the Ndjolé area, in central Ga...Innspub Net
The forests of the Congo Basin cover an area of 200 million hectares, of which just over 10% is in Gabon. In this country, crop products and non-timber forest products (NTFPs) are abundant because of its favourable climate. There is significant biodiversity and great potential for non-timber forest products. This study is interested in the identification and the supply chain of the Marantaceae, one of these NTFPs of plant origin in central Gabon, whose exploitation is national. Through a survey of the main actors in the sector and field visits in the locality of Bifoun, it emerges the existence of three large groups of exploited marantaceae, whose harvest and transport are mainly done by women, in various containers. The storage of this NTFPs does not exceed four days, with the risk of losing its commercial value due to drying out. The uses of this resource are multiple: processing cassava, cooking food, making handicrafts, etc. The income it provides to producers is mainly use towards small family expenses but helps to monetize the rural world. The difficulties inherent in the distance from harvesting points, the low price of the product and the impossibility of long-term storage of the marantaceae leaves constitute a brake on the development of this activity. It would therefore be wise to envisage the domestication of the species listed for a sustainable use of this plant genetic resource.
Identification and marketing of Marantaceae in the Ndjolé area, in central GabonOpen Access Research Paper
The forests of the Congo Basin cover an area of 200 million hectares, of which just over 10% is in Gabon. In this country, crop products and non-timber forest products (NTFPs) are abundant because of its favourable climate. There is significant biodiversity and great potential for non-timber forest products. This study is interested in the identification and the supply chain of the Marantaceae, one of these NTFPs of plant origin in central Gabon, whose exploitation is national. Through a survey of the main actors in the sector and field visits in the locality of Bifoun, it emerges the existence of three large groups of exploited marantaceae, whose harvest and transport are mainly done by women, in various containers. The storage of this NTFPs does not exceed four days, with the risk of losing its commercial value due to drying out. The uses of this resource are multiple: processing cassava, cooking food, making handicrafts, etc. The income it provides to producers is mainly use towards small family expenses but helps to monetize the rural world. The difficulties inherent in the distance from harvesting points, the low price of the product and the impossibility of long-term storage of the marantaceae leaves constitute a brake on the development of this activity. It would therefore be wise to envisage the domestication of the species listed for a sustainable use of this plant genetic resource.
Genotoxicity of Eleusine indica (Nkim enang: Efik) was investigated in the Wister strain albino rat (Rattus novergicus). Nine (9) male and nine (9) female rats were randomly assigned to three (3) groups, of which two were exposed to the aqueous extract of E.indica – Group A (control-no extract)), Group B (50 mg/kg BW of E. indica) and Group C (100 mg/kg BW of E. indica). This was administered to the rats by oral gavage for 14 days after which the peripheral blood from the tail tips were collected and assayed for the presence of micronuclei, following standard procedures. Proximate analysis and phytochemical screening of the herb extract was carried out. Results obtained showed that E. indica did not cause any significant (P > 0.05) increase in the incidence of micronucleated polychromatic erythrocytes in rat peripheral blood at any of the doses administered. The polychromatic: normochromatic erythrocyte (PCE: NCE) ratio was found to be in the range of 0.50 ± 0.11 to 0.55 ± 0.02. Also, the aqueous herb extract is rich in Carbohydrates (76.17%) and Tannins (21.76%). Mean body weights (MBW) of rats showed normal distribution throughout the duration of the investigation. The results of this study demonstrate that E. indica does not confer any genotoxicity in mammals. Further in-depth study on its efficacy is recommended.
2. 314 Journal of Nematology, Volume 37, No. 3, September 2005
TABLE 1. Population code, origin, soil type, enzyme phenotype, and percentage occurrence of Meloidogyne spp. on “Cavendish” bananas
from Martinique.
Enzyme phenotypeb
Population Occurrencec
code Township Soil type Species Est Mdh %
1 Saint Joseph Halloysitic M. arenaria A2 N1 100
2 Saint Joseph Halloysitic M. incognita I2 N1 100
3A Saint Joseph Halloysitic M. arenaria A2 N1 80
3B Halloysitic M. incognita I2 N1 20
4 Carbet Allophanic M. arenaria A2 N1 100
5 Carbet Allophanic M. incognita I2 N1 100
6A Carbet Allophanic M. arenaria A2 N1 11.7
6B Allophanic M. incognita I2 N1 88.3
7A Carbet Allophanic M. arenaria A2 N1 50
7B Allophanic M. incognita I2 N1 50
8 Aa Carbet Allophanic M. incognita I2 N1 38.4
8 Ba Allophanic Meloidogyne sp. B2 N1 7.8
8C Allophanic M. hispanica Hi3 N3 53.8
9A Carbet Allophanic M. arenaria A2 N1 45.2
9B Allophanic M. arenaria A2 N3 32.3
9C Allophanic M. incognita I2 N1 22.5
10 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
11 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
12 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
13 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
14a Ajoupa Bouillon Andosol M. arenaria A2 N1 100
15 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
16 Ajoupa Bouillon Andosol M. arenaria A2 N1 100
17 Sainte Marie Ferrisol M. arenaria A2 N1 100
18 Sainte Marie Ferrisol M. arenaria A2 N1 100
19 Sainte Marie Ferrisol M. incognita I2 N1 100
20 Gros-Morne Halloysitic M. arenaria A2 N1 100
21 Gros-Morne Halloysitic M. arenaria A2 N1 100
22 Trinite´ Ferrisol M. arenaria A2 N1 100
23 Trinite´ Ferrisol M. arenaria A2 N1 100
24 Trinite´ Ferrisol M. incognita I2 N1 100
25 A Trinite´ Ferrisol M. arenaria A2 N1 60
25 B Trinite´ Ferrisol M. incognita I2 N1 40
26 Francois
¸ Ferrisol M. arenaria A2 N1 100
27 Francois
¸ Ferrisol M. arenaria A2 N1 100
28 Francois
¸ Ferrisol M. arenaria A2 N3 100
29 A Francois
¸ Ferrisol M. arenaria A2 N1 55
29 B Ferrisol M. arenaria A2 N3 45
30 A Francois
¸ Ferrisol M. arenaria A2 N1 80
30 B Ferrisol M. incognita I2 N1 20
31 A Francois
¸ Ferrisol M. arenaria A2 N1 40
31 B Ferrisol M. incognita I2 N1 60
32 Vauclin Vertisol M. arenaria A2 N1 100
33 Vauclin Vertisol M. arenaria A2 N1 100
34 Vauclin Vertisol M. arenaria A2 N1 100
35 Vauclin Vertisol M. incognita I2 N1 100
36 A Vauclin Vertisol M. arenaria A2 N1 34.5
36 B Vertisol M. arenaria A2 N3 65.5
37 A Vauclin Vertisol M. arenaria A2 N1 89
37 B Vertisol M. incognita I2 N1 11
38 Lamentin Alluvial M. arenaria A2 N1 100
39 A Saint-Pierre Allophanic M. incognita I2 N1 100
40 Saint-Pierre Allophanic M. incognita I2 N1 100
41 Saint-Pierre Allophanic M. incognita I2 N1 100
42 Saint-Pierre Allophanic M. incognita I2 N1 100
43 A Saint-Pierre Allophanic M. arenaria A2 N1 52.4
43 B Allophanic M. incognita I2 N1 47.6
44 Basse Pointe Allophanic M. incognita I2 N1 100
45 Basse Pointe Allophanic M. incognita I2 N1 100
46 Basse Pointe Allophanic M. incognita I2 N1 100
47 A Basse Pointe Allophanic M. arenaria A2 N1 35.7
47 B Allophanic M. incognita I2 N1 64.3
48 A Marigot Halloysitic M. incognita I2 N1 100
49 A Marigot Halloysitic M. arenaria A2 N1 50
49 B Halloysitic M. arenaria A2 N3 50
3. Meloidogyne spp. on Musa: Cofcewicz et al. 315
TABLE 1. Continued
Enzyme phenotypeb
Population Occurrencec
code Township Soil type Species Est Mdh %
50 Macouba Allophanic M. incognita I2 N1 100
51 Macouba Allophanic M. incognita I2 N1 100
52 A Macouba Allophanic M. arenaria A2 N1 28.6
52 B Allophanic M. incognita I2 N1 71.4
53 A Macouba Allophanic M. incognita I2 N1 80
53 B Allophanic M. incognita I1 N1 20
54 A Sainte-Anne Vertisol M. arenaria A2 N1 20
54 B Vertisol M. incognita I2 N1 80
55 A Sainte-Anne Vertisol M. arenaria A2 N1 81.8
55 B Sainte-Anne Vertisol M. incognita I2 N1 18.2
a
Meloidogyne spp. isolates used in morphological and DNA analyses.
b
Phenotype designations: Est = esterase, Mdh = malate dehydrogenase.
c
Percentage of occurrence of Meloidogyne spp. phenotypes in 10 analyzed females.
analyzed to obtain isozyme profiles of esterase (Est) in glycerin (Taylor and Netscher, 1974), and at least 30
and malate dehydrogenase (Mdh) (Esbenshade and specimens were examined from each isolate.
Triantaphyllous, 1985, 1990). Electrophoresis was per- For molecular analysis, the eggs of each isolate were
formed with PhastGel gradient 10% to 15%, in discon- extracted using the method of Cofcewicz et al. (2004).
tinuous buffer system using an automated system Total genomic DNA was extracted from 150 to 200 µl of
(PhastSystem, GMI, Inc., Ramsey, MN). Ten young fe- nematode eggs of each available isolate that had been
males were collected directly from the banana root stored at −80°C. Eggs were placed in liquid nitrogen
sample and individually macerated in 0.6 µl of extrac- and ground with a pestle and mortar. The DNA was
tion buffer containing 20% sucrose, 2% Triton X-100, purified from the resulting powder by a phenol-
and 0.01% Bromophenol blue. The macerate was ap- chloroform extraction (Sambrook et al., 1989). Follow-
plied with two combs in the electrophoresis apparatus ing ethanol precipitation, DNA was resuspended in TE
(Karssen et al., 1995). On each gel slab the protein buffer.
extracts of two females of M. javanica (Treub) Chit- Forty-one random 10-mer primers, purchased from
wood were included as a reference standard. Eurogentec (Herstal, Belgium), were used in RAPD ex-
Gels were stained for esterase activity for 60 min, periments (Table 4). RAPD-PCR was performed in a
and for malate dehydrogenase for 10 min (Esbenshade final volume of 25 µl containing 5 ng of total genomic
and Triantaphyllou, 1985). Gels were washed and DNA; 80 pM of primer, dATP, dCTP, dGTP, and dTTP
placed in a solution containing 10% acetic acid, 8% each at 200 µM final concentration; 1xTaq incubation
glycerol, and 82% water for 1 day. Gels were pressed buffer; and 1.25 units Taq polymerase (Phoneutria Bio-
between two cellophane sheets and left to dry (Car- technologia & Servicos, SP, Brazil). Amplification was
¸
neiro and Almeida, 2001). Enzyme phenotypes were performed on a PTC-100MJ Research thermal cycler
designated with letter(s) suggestive of the species and (MJ Research Inc., Waltham, MA). The cycling pro-
the number of bands (Esbenshade and Triantaphyllou, gram was 1 min at 94 °C, 40 cycles of 20 sec at 94 °C, 30
1985, 1990). sec at 36 °C, and 2 min at 70 °C. Amplification products
Due to limitations in the biological material avail- were separated by electrophoresis in 1.4% agarose gels
able, the RAPD and morphological analyses were per- in TBE buffer at a constant current of 150 mA for ap-
formed only in 16 isolates, which were single egg mass proximately 3 hours and visualized with ethidium bro-
isolates and transported to Brazil (Embrapa Recursos mide (0.5 µg ml−1) under UV light.
Geneticos e Biotecnologia, Brasılia, DF) to complete
´ ´ Observed DNA bands were scored as present or ab-
the analyses. These comprised two isolates of M. ja- sent directly from the gels. For each isolate, two inde-
vanica, two isolates of M. cruciani Garcia-Martinez, Tay- pendent PCR reactions were electrophoresed in the
lor & Smart, 1982, seven isolates of M. arenaria (Neal) same gel; only DNA fragments consistently present or
Chitwood, three isolates of M. incognita (Kofoid & absent in these replicated samples were scored as bi-
White) Chitwood, and two isolates of an unidentified nary characters. Each reaction was repeated at least
Meloidogyne species. Unfortunately, the isolates of M. once. DNA fingerprints from each isolate were con-
hispanica Hirschmann, M. incognita (I2N3), and the iso- verted to a 0-1 matrix and a phylogenetic analysis was
late Meloidogyne sp. (B0N1) without esterase bands were conducted using the computer program PAUP*4.0
lost during the transportation to Brazil (Tables 1, 2, 3). (Swofford, 1998). Distance analysis was performed ac-
For the morphological studies, perineal patterns cording to the UPGMA method. One thousand boot-
were cut from females in 45% lactic acid and mounted strap replicates were performed to test the node sup-
4. 316 Journal of Nematology, Volume 37, No. 3, September 2005
TABLE 2. Population code, origin, soil type, enzyme phenotype, and percentage occurrence of Meloidogyne spp. on “Cavendish” bananas
from Guadeloupe.
Enzyme
phenotypeb
Population Occurrencec
code Township Soil type Species Est Mdh %
1 Saint-Caude Andosol M. arenaria A2 N1 100
2 Saint-Caude Andosol M. arenaria A2 N1 100
3 Saint-Caude Andosol M. arenaria A2 N1 100
4 Goyave Ferrisol M. arenaria A2 N1 100
5A Goyave Ferrisol M. arenaria A2 N1 78.9
5 Ba Ferrisol M. arenaria A2 N3 21.1
6A Goyave Ferrisol M. arenaria A2 N1 82.3
6B Ferrisol M. incognita I2 N1 17.7
7 Trois-Rivières Andosol M. arenaria A2 N1 100
8A Trois-Rivières Andosol M. arenaria A2 N1 81.8
8B Andosol M. incognita I2 N1 18.2
9A Trois-Rivières Andosol M. arenaria A2 N1 9.9
9B Andosol M. incognita I2 N1 90.1
10 Baie-Mahault Ferrisol M. arenaria A2 N1 100
11a Capesterre-Belle-Eau Halloysitic M. arenaria A2 N1 100
12 Capesterre-Belle-Eau Andosol M. arenaria A2 N1 100
13 Capesterre-Belle-Eau Andosol M. arenaria A2 N1 100
14 Capesterre-Belle-Eau Andosol M. arenaria A2 N1 100
15 Capesterre-Belle-Eau Alluvial M. arenaria A2 N1 100
16 Capesterre-Belle-Eau Andosol M. arenaria A2 N1 100
17 A Capesterre-Belle-Eau Andosol M. arenaria A2 N1 70.6
17 B Andosol M. incognita I2 N1 29.4
18 A Capesterre-Belle-Eau Andosol M. arenaria A2 N1 68.2
18 Ba Andosol M. cruciani Cr3 N1 31.8
19 A Capesterre-Belle-Eau Andosol M. arenaria A2 N1 72.7
19 Ba Andosol Meloidogyne sp. Ba2 N1 27.3
20 Capesterre-Belle-Eau Andosol Meloidogyne sp. B0 N1 100
21 Petit-Canal Vertisol M. arenaria A2 N1 100
22 Bouillante Ferrisol M. arenaria A2 N1 100
23a Vieux-Habitants Ferrisol M. arenaria A2 N1 100
24 A Vieux-Habitants Alluvial M. arenaria A2 N1 66.7
24 B Alluvial M. incognita I2 N1 33.3
25 A Vieux-Habitants Alluvial M. arenaria A2 N1 81.8
25 Ba Alluvial M. incognita I2 N1 18.2
26 Saint-Francois
¸ Vertisol M. incognita I2 N1 100
27 A Petit Bourg Ferrisol M. arenaria A2 N1 27.3
27 B Ferrisol M. incognita I2 N1 72.7
28 A San Julian Vertisol M. incognita I2 N1 13.3
28 B Vertisol M. arenaria A2 N1 60
28 Ca Vertisol M. arenaria A1 N1 26.7
a
Meloidogyne spp. isolates used in morphological and DNA analyses.
b
Phenotype designation: Est = esterase, Mdh = malate dehydrogenase.
c
Percentage of occurrence of Meloidogyne spp. phenotypes in 10 analyzed females.
port (Felsenstein, 1985), and a consensus dendrogram and 3 populations of M. incognita, respectively, from
was computed. Martinique (Table 1; Figs. 1, 2) and French Guiana
(Table 3; Figs. 1, 2). In Guadeloupe nine populations
Results with the phenotype I2 were detected, but the pheno-
type I1 was not observed (Table 2; Figs. 1, 2).
Esterase characterization: Eleven bands for Est activity The phenotype A2 (Rm 1.2, 1.3) was detected in 41
were detected in the 96 Meloidogyne populations. A dis- populations of M. arenaria from Martinique (Table 1;
tinct Est-phenotype was associated with the major and Figs. 1, 2), in 27 populations from Guadeloupe (Table
minor Meloidogyne species (Tables 1, 2, 3, 4; Figs. 1, 2). 2; Figs. 1, 2), and in nine populations from French
The species-specific phenotype J3 (Rm: 1.0, 1.25, 1.4) Guiana (Table 3; Figs. 1, 2). Only one population with
and phenotype J2 (Rm: 1.0, 1.25) were detected in two the phenotype A1 (Rm 1.2) was detected from Guade-
M. javanica populations from French Guiana (Table 3; loupe (Table 2; Figs. 1, 2).
Figs. 1, 2). The phenotype Cr3 of M. cruciani with three bands
The phenotype I2 (Rm 1.05), with a minor band (Rm (Rm 1.05, 1.28, 1.38) was detected in two populations
1.10), and phenotype I1 (Rm 1.0) were detected in 39 from Guadeloupe and French Guiana.
5. Meloidogyne spp. on Musa: Cofcewicz et al. 317
TABLE 3. Population code, origin, soil type, enzyme phenotype, and percentage occurrence of Meloidogyne spp. on “Cavendish” bananas
from French Guiana.
Enzyme
phenotypesb
Population Soil Occurrencec
code Township type Species Est Mdh %
1 Aa Macouria Alluvial M. incognita I2 N1 50
1 Ba Alluvial M. cruciani Cr3 N3 40
1 Ca Alluvial M. javanica J3 N1 10
2 Macouria Alluvial M. incognita I2 N1 100
3A Macouria Alluvial M. incognita I2 N1 80
3 Ba Alluvial M. arenaria A2 N3 20
4A Macouria Alluvial M. arenaria A2 N3 90.9
4 Ba Alluvial M. javanica J2 N1 9.1
5A Macouria Alluvial M. arenaria A2 N3 93.3
5B Alluvial M. incognita I2 N1 6.7
6A Macouria Alluvial M. arenaria A2 N1 27.3
6B Alluvial M. incognita I2 N1 72.7
7 Regina
´ Ferrisol M. incognita I1 N1 100
8 Regina
´ Ferrisol M. arenaria A2 N1 100
9A Cacao Ferrisol M. arenaria A2 N1 27.3
9B Ferrisol M. arenaria A2 N3 72.7
10 A Cacao Ferrisol M. incognita I2 N1 33.3
10 B Ferrisol M. arenaria A2 N1 66.7
11 A Counamama Alluvial M. incognita I1 N1 10
11 B Alluvial M. incognita I2 N1 90
12 Counamama Alluvial M. incognita I2 N1 100
13a Rocoucoua Ferrisol M. arenaria A2 N1 100
a
Meloidogyne spp. isolates used in morphological and DNA analyses.
b
Phenotype designations: Est = esterase, Mdh = malate dehydrogenase.
c
Percentage of occurrence of Meloidogyne spp. phenotypes in 10 analyzed females.
The phenotype Ba2 (Rm 1.05, 1.38) was detected in specific and were designated by the letter N for non-
one population from Guadeloupe and is a new esterase specific phenotype and a number indicating the num-
phenotype. ber of bands of activity.
In Martinique, the phenotype Hi3 (Rm 0.8, 0.9, 1.05) The Mdh N1 (Rm 1.0) and N3 (Rm 1.0, 1.1, 1.2)
of M. hispanica (Hirschmann, 1986) was detected differentiated two populations of M. arenaria. Pheno-
mixed with one population of Meloidogyne sp., Est phe- type N1 was detected in 69, 39, and 17 populations
notype B2 (Rm 0.9, 1.05). One population (Meloidogyne from Martinique, Guadeloupe, and French Guiana, re-
sp.) from Guadeloupe (Table 2) did not give any major spectively. Phenotype N3 was detected in six, one, and
esterase bands (B0). five populations.
In the banana field conditions of Martinique, M. are- The populations of M. incognita were differentiated
naria, M. incognita, and Meloidogyne sp., were detected in for Mdh phenotypes in French Guiana, where pheno-
58.8%, 40.1%, and 1.1%, respectively. Of the 55 ana- type N1 appeared in 10 populations and N3 in one
lyzed areas, 34.6% presented multiple species and the population. All the minor species, except M. hispanica
other 65.4% individual species (Table 1). (N3), had the phenotype N1.
In Guadeloupe, M. arenaria, M. incognita, and Morphological features: The M. arenaria perineal pat-
Meloidogyne spp. were detected in 80.9%, 13.4%, and terns as described by Eisenback and Triantaphyllou
5.7% of the samples, respectively. Of the 28 locations (1991) were observed in the isolates of phenotypes
sampled, 39.3% contained multiple species and the A2N1, A1N1. The M. arenaria isolated with phenotypes
other 60.7% contained an individual species (Table 2). A2N3 had the different perineal pattern reported pre-
In French Guiana, M. arenaria, M. incognita, and viously by Cliff and Hirschmann (1985). Dorsal arch
Meloidogyne spp. were detected in 46.0%, 49.4%, and was usually high and rounded to squarish in shape.
4.6%, respectively. Of the 13 locations sampled, 61.5% Occasionally dorsal striae form shoulders in lateral re-
contained multiple species and the other 38.5% con- gions of the pattern. Lateral lines near the tail tip were
tained an individual species, with M. arenaria and M. often widely spaced, with few broken irregular striae
incognita prevailing (Table 3). between lines. Anteriorly from the tail tip lines became
Malate Dehydrogenase (Mdh): Three bands of Mdh ac- finer. Dorsal and ventral striae were wavy, irregular, or
tivity were detected in the 96 populations of RKN stud- forked near lateral lines but usually smooth thoughout
ied (Fig. 1). Two distinct phenotypes were recognized remainder of the pattern, never a forming zigzag pat-
(Fig. 1). However, the phenotypes were not species- tern (Cliff and Hirschmann, 1985).
6. 318 Journal of Nematology, Volume 37, No. 3, September 2005
TABLE 4. RAPD primers used and number of amplified fragments scored.
N° amplified fragments
Primer 5Ј sequence 3Ј % GC Minimum/ isolate Maximum/isolate Polymorphic Total
A01 CAG GCC CTT C 70 7 14 28 28
A04 AAT CGG GCT G 60 3 15 22 24
A12 TCG GCG ATA G 60 5 8 17 17
B11 GTA TAC CCG T 50 4 10 13 15
B12 CCT TGA CGC A 60 7 12 13 14
B17 AGG GAA CGA G 60 3 6 9 10
C02 GTG AGG CGT C 70 4 10 20 20
C07 GTC CCG ACG A 70 2 12 22 23
C09 CTC ACC GTC C 70 5 8 9 12
C16 CAC ACT CCA G 60 3 9 17 18
D05 TGA GCG GAC A 60 8 14 16 18
D13 GGG GTG ACG A 70 9 14 17 21
E07 AGA TGC AGC C 60 8 17 17 22
G02 GGC ACT GAG G 70 6 17 21 23
G03 GAG CCC TCC A 70 2 5 8 9
G05 CTG AGA CGG A 60 5 13 23 24
G06 GTG CCT AAC C 60 1 12 20 21
G13 CTC TCC GCC A 70 7 13 18 20
J10 AAG CCC GAG G 70 5 13 24 25
J20 AAG CGG CCT C 70 5 12 14 16
K01 CAT TCG AGC C 60 4 12 17 18
K04 CCG CCC AAA C 70 10 22 38 40
K06 CAC CTT TCC C 60 9 16 23 24
K10 CAC CTT TCC C 60 4 12 17 20
K14 CCC GCT ACA C 70 4 7 15 15
K16 GAG CGT CGA A 60 7 12 19 21
K19 CAC AGG CGG A 70 9 14 18 20
K20 GTG TCG CGA G 70 9 18 29 30
L08 AGC AGG TGG A 60 8 13 18 21
M20 AGG TCT TGG G 60 8 16 27 29
N07 CAG CCC AGA G 70 4 10 11 12
P01 GTA GCA CTC C 60 6 16 24 26
P02 TCG GCA CGC A 70 8 11 12 16
PO5 CCC CGG TAA C 70 5 12 16 17
R05 GAC CTA GTG G 60 9 15 22 23
R07 ACT GGC CTG A 60 6 12 21 21
R08 CCC GTT GCC T 70 11 13 28 29
AB02 GGA AAC CCC T 60 9 14 31 32
AB04 GGC ACG GGT T 70 7 16 22 24
AB06 GTG GCT TGG A 60 6 16 29 30
AB09 GGG CGA CTA C 70 10 22 31 33
Total - - 816 881
The phenotype I2N1 had perineal patterns typical of have wing-like striae on one or both sides, a well-
M. incognita and the two isolates of M. javanica, pheno- defined tail tip area with striations in most specimens,
types J3N1 and J2N1, had perineal patterns like those and an anus distinct and often covered by a thick cu-
described for the species (Eisenback and Triantaphyl- ticular fold.
lou, 1991). RAPD characterization: With the 41 random primers
The phenotype Cr3N1 had perineal patterns with used, the number of reproducible fragments varied
subcuticular punctuations surrounding the anus on the from 1 to 22/isolate, and ranged from ca 200 to ca
lateral and posterior side; striae deep, wavy, sometimes 4,000 bp in size. One gel containing the 16 isolate pat-
broken; lateral field deep with distinct phasmids; and terns obtained with primer E07 is shown in Figure 3.
tail terminus indistinct, as described for M. cruciani. The global results of the RAPD analysis are provided in
(Garcıa-Martinez et al., 1982). Some perineal patterns
´ Table 4. Over the entire experiment, each primer pro-
of this species resembled those of M. javanica in having duced from eight to 38 polymorphic bands. Overall,
distinct lateral lines; however, the lateral lines of M. 881 fragments were amplified and scored as RAPD
cruciani did not extend as far as those of M. javanica. markers, 816 of which were polymorphic, and 65 were
The phenotype B2N1 had perineal patterns elon- found to be amplified in all the isolates tested (i.e.,
gated to ovoid with a flattened-to-high, squarish dorsal monomorphic).
arch with widely spaced coarse striae. Lateral fields may For the four species for which more than one isolate
7. Meloidogyne spp. on Musa: Cofcewicz et al. 319
clustered together with 62% of bootstrap support and
57.4% of polymorphism; (iv) all isolates of M. arenaria
(A2N1, A1N1, and A2N3) were clustered together, but
the support for their clustering was much lower than
that for the other species (52% in the bootstrap); (v)
M. arenaria isolates phenotypes (A2N1 and A1N1) clus-
tered together with a bootstrap of 100%; (vi) the atypi-
cal isolate phenotype Ba2N1 (19 B Gua) clustered to-
gether with M. arenaria (A2N1, A1N1) with a bootstrap
of 92%, but with polymorphism of 53.7%; (vi) M. are-
naria isolates A2N3 clustered together with a bootstrap
of 100%; (vii) all isolates of M. incognita clustered to-
gether (100% of bootstrap) and with the other atypical
isolate phenotype B2N1 with a high confidence level
(100% of bootstrap), but with a polymorphism of
62.4%.
Discussion
Fig. 1. Phenotypes of esterase (Est) and malate dehydrogenase This study presents a characterization of Meloidogyne
(Mdh) observed in 96 populations of Meloidogyne spp. from Musa. species collected on banana fields in Martinique,
(For origin of populations, see Tables 1, 2, and 3.)
Guadeloupe, and French Guiana, using isozyme phe-
notype (Est and Mdh), morphology, and DNA ap-
was available (i.e., M. arenaria, M. incognita, M. javanica, proaches. A total of 96 banana fields were sampled and
and M. cruciani), 307 to 576 reproducible fragments analyzed. This was the most detailed and extensive RKN
were amplified (Table 5). For M. incognita, M. javanica, survey to have been carried out in the French Overseas
and M. cruciani, fewer RAPD bands were polymorphic Departments.
(14.9%, 16.6%, and 16.1%, respectively). For M. are- Results confirm that the use of esterase phenotypes is
naria, if all esterase phenotypes are placed in a single a rapid and efficient method to i) characterize Meloido-
group (Table 5), 61.6% of the amplified bands were gyne species and detect atypical forms, ii) determine the
polymorphic. If the different esterase phenotypes were frequency and relative distribution of Meloidogyne spp.
separated into sub-groups (A2N1, A1N1 + A2N1 and in field surveys, and iii) detect mixed species popula-
A2N3), then 29%, 37.3% and 23.2%, respectively, of tions prior to conducting subsequent studies such as
the bands were polymorphic. DNA analyses and morphological characterization
In the dendrogram of relationships between the 16 (Carneiro et al., 1996, 2000; Cofcewiz et al., 2004; Es-
isolates based on the RAPD polymorphisms, the isolates benshade and Triantaphyllou, 1985, 1990).
belonging to a given species always clustered together Malate dehydrogenase phenotypes detected intraspe-
(Fig. 4). These results confirmed the existence of three cific variability only among isolates of M. arenaria and
separate groups corresponding to the different species: M. incognita. A similar result was observed among popu-
(i) the two isolates of M. javanica clustered together lations of Meloidogyne from Portugal (Pais and Abrantes,
with a bootstrap support of 100% in the analysis; (ii) 1989).
the two isolates of M. cruciani clustered together (boot- The perineal pattern is often an unreliable character
strap support of 100%); (iii) M. javanica and M. cruciani for making diagnostic conclusions when used alone
but, when used as a complement to isozyme pheno-
types, species identification is more robust (Carneiro et
al., 2004). The RAPD polymorphism data were consis-
tent with isozyme phenotype data and morphology for
identification of species and estimating genetic rela-
tionship and diversity among isolates (Carneiro et al.,
2004; Cofcewicz et al. 2004).
Meloidogyne arenaria was the most common species on
banana in Martinique, Guadeloupe, and French Gui-
ana. The phenotypes A3, A2, and A1 were species-
specific for M. arenaria (Esbenshade and Triantaphyl-
lou, 1985, 1990), with the A2N1 phenotype being most
Fig. 2. Eleven esterase phenotypes found in 96 populations of
Meloidogyne spp. (For explanation of phenotype designations, see common. The phenotype A1N1 detected only once in
Fig. 1.) Guadeloupe and the A2N3 phenotype was intermediate
8. 320 Journal of Nematology, Volume 37, No. 3, September 2005
Fig. 3. RAPD patterns for 16 Meloidogyne spp. isolates generated with primer E07. For each isolate, two duplicate amplifications were loaded
side by side on the gel. (C: control reaction without template DNA. M: Nucleic acid marker in bp. Population codes are given in Tables 1, 2,
and 3.)
in occurence and was detected in Martinique, Guade- possibly a species complex. Alternatively, these varia-
loupe, and French Guiana. The A3N1 phenotypes did tions may indicate that M. arenaria is an old species that
not appear during this survey but were detected from has diverged considerably through multiple mutations
banana in Brazil (Cofcewicz et al., 2004). The M. are- and adaptations to different environments (Esben-
naria isolates with phenotype A2N1 clustered with 99% shade and Triantaphyllou, 1987).
of bootstrap and separated from the isolates A1N1 and The atypical population with the Ba2N1 had a new
A2N3, showing a very high intraspecific variability esterase phenotype not previously detected. This popu-
based on DNA analysis and congruent with isozyme lation also had a perineal pattern similar to those of M.
phenotypes. These results agree with the previous mo- arenaria and in RAPD analysis clustered with the group
lecular analysis on the genome (Castagnone—Sereno M. arenaria with high polymorphism. Additional studies
et al., 1994; Cofcewicz et al., 2004; Randig et al., 2002; will be necessary to identify this isolate.
Semblat et al., 1998). The isolates A2N1 and A1N1 pre- The Caribbean populations of M. incognita had two
sented perineal patterns like those described for M. esterase phenotypes: I2 and occasionally I1. The same
arenaria, but the isolates with phenotype A2N3 had dif- phenotypes were observed by Pais and Abrantes (1989)
ferent perineal patterns, as previously demonstrated and Carneiro et al. (1996, 2000). Phenotype I1 was also
(Cliff and Hirschmann, 1985). Meloidogyne arenaria is rarely observed among populations from soybean
known to be morphologically, physiologically, and cy- (Castro et al., 2003) or banana (Cofcewicz et al., 2004)
togenetically variable (Cliff and Hirschmann, 1985; Tri- in Brazil. As observed in this study, Carneiro et al.
antaphyllou, 1985). This variation may indicate mul- (2004), using RAPD analysis, showed that I1 and I2
tiple origins for populations classified as M. arenaria or clustered together and had relatively low polymor-
TABLE 5. Rate of RAPD polymorphisms observed at species level.
RAPD fragments
Species Amplified Polymorphic (%)
M. javanica (J3N1, J2N1) (Isolates: 1C Gui, 4B Gui) 307 51 (16.6%)
M. cruciani (Cr3N1) (Isolates: 1B Gui, 18B Gua) 378 61 (16.1%)
M. javanica + M. cruciani (1C Gui, 4B Gui, 1B Gui, 18B Gua) 467 268 (57.4%)
M. incognita (I2N1) (Isolates: 25 B Gua, 8A Mar, 1 A Gui) 401 60 (14.9%)
M. incognita + Meloidogyne sp. (8B Mar) 556 347 (62.4%)
M. arenaria (A2N1) (Isolates: 23 Gua, 11 Gua, 13 Gui, 14 Mar) 382 111 (29.0%)
M. arenaria (A2N3) 5B Gua, 3B Gui 379 88 (23.2%)
M. arenaria (A2N1 + A1N1) (Isolates: 23 Gua, 11 Gua, 13 Gui, 14 Mar, 28C Gua) 421 157 (37.3%)
M. arenaria (A2N1 + A1N1) + Meloidogyne sp. (Ba2N1) (Isolate: 19B Gua) 497 267 (53.7%)
M. arenaria group (A2N1 + A1N1 + A2N3) (Isolates: 23 Gua, 11 Gua, 13 Gui, 14 Mar, 28C Gua, 5B Gua,
3B Gui) 513 316 (61.6%)
M. arenaria + Meloidogyne sp1(A2N1 + A1N1 + A2N3 + Ba2N1) (Isolates: 23 Gua, 11 Gua, 13 Gui, 14 Mar,
28C Gua, 5B Gua, 3B Gui, 19B Gua) 576 405 (70.3%)
9. Meloidogyne spp. on Musa: Cofcewicz et al. 321
ana and Guadeloupe, also is a minor species presenting
a species-specific esterase phenotype (Cr3) character-
ized first as M3a by Esbenshade and Triantaphyllou
(1985). Based on RAPD data, M. javanica and M. cru-
ciani were related species; the similarities in perineal
patterns also suggest a close relationship between M.
cruciani and M. javanica (Garcia-Martinez et al., 1982).
One population from Martinique exhibited the phe-
notype Hi3 similar to that found in M. hispanica and
referred to as ЈЈSeville populations’’ (Dalmasso and
Berge, 1978; Janati et al. 1982) and later designated as
´
S2-M1 (Esbenshade and Triantaphyllou, 1985). Two
Mdh phenotypes, N1 and N3, were observed for differ-
ent populations of M. hispanica (Esbenshade and Tri-
antaphyllou, 1985) and for Portuguese populations
(Pais and Abrantes, 1989), respectively. Our population
had the same phenotype, N3, as detected for Portu-
guese isolates.
The lack of esterase bands (Est B0) was observed first
by Esbenshade and Triantaphyllou (1985). The same
phenotype was described for M. fallax Karssen (1996),
but the Mdh is not N1 and is specific for this species.
The identification of Meloidogyne species parasitizing
bananas needs to be more accurate. Our data indicate
the value of using a combination of morphological and
biochemical traits for species identification.
Fig. 4. Majority-rule consensus UPGMA dendrogram of relation-
ships of 16 Meloidogyne spp. isolates (M. javanica: jav, M. cruciani: cru, Literature Cited
M. arenaria: are, M. incognita: inc, Meloidogyne sp.: sp) based on RAPD Baum, T. J., P. M. Gresshoff, S. A. Lewis, and R. A. Dean. 1994.
data. (Bootstrap percentages based on 1,000 replicates are given on Characterization and phylogenetic analysis of four root-knot nema-
each node. Population codes are given in Tables 1, 2, and 3.) tode species using DNA amplification fingerprinting and automated
polyacrylamide gel electrophoresis. Molecular Plant Microbe Inter-
action 7:39–47.
phism, despite the populations having different ester- Blok, V. C., M. S. Phillips, J. W. McNicol, and M. Fargette. 1997.
ase phenotypes. Genetic variation in tropical Meloidogyne spp. as shown by RAPDs.
The atypical population of Meloidogyne sp. (pheno- Fundamental and Applied Nematology 20:127–133.
Carneiro, R. M. D. G., and M. R. A. Almeida. 2001. Tecnica de
´
type B2N1) from Martinique has also been found in eletroforese usada no estudo de enzimas dos nemato ´ides de galhas
sugar-cane (Saccharum spp.) in the northeast part of the para identificacao de especies. Nematologia Brasileira 25:35–44.
¸˜ ´
island (Queneherve, pers. comm.). The RAPD data
´ ´ ´ Carneiro, R. M. D. G., M. R. A. Almeida, and R. G. Carneiro. 1996.
clustered this population with M. incognita isolates but Enzyme phenotypes of Brazilian isolates of Meloidogyne spp. Funda-
mental and Applied Nematology 19:555–560.
with sufficient polymorphism that it could not be iden- Carneiro, R. M. D. G., M. R. A. Almeida, and P. Queneherve. 2000.
´ ´ ´
tified as M. incognita. The esterase phenotype B2N1 Enzyme phenotypes of Meloidogyne spp. populations. Nematology 2:
(labelled S1-M1) has also been reported from Nigeria, 645–654.
the Cote-d’Ivoire, the Philippines, Samoa (Fargette, Carneiro, R. M. D. G., P. Castagnone-Sereno, and D. W. Dickson.
1998. Variability among four populations of Meloidogyne javanica from
1987), and the United States (Esbenshade and Trian- Brazil. Fundamental and Applied Nematology 21:319–326.
taphyllou, 1985). Carneiro, R. M. D. G., M. S. Tigano, O. Randig, M. R. A. Almeida,
During this survey, M. javanica was rarely found on and J. L. Sarah. 2004. Identification and genetic diversity of Meloido-
gyne spp. (Tylenchida: Meloidogynidae) on coffee from Brazil, Cen-
banana but was commonly found parasitizing Musa in
tral America, and Hawaii. Nematology 6:37–47.
Brazil (Cofcewicz et al., 2004). Most of the M. javanica Castagnone-Sereno, P., F. Vanlerberghe-Masutti, and F. Leroy.
populations have a single esterase phenotype, J3, not 1994. Genetic polymorphism between and within Meloidogyne species
found in any other Meloidogyne species (Esbenshade detected with RAPD markers. Genome 37:904–909.
Castro, J. M. C. E., R. D. Lima, and R. M. D. G. Carneiro. 2003.
and Triantaphyllou, 1990). The phenotype J2 was first
Variabilidade isoenzimatica de populacoes de Meloidogyne spp. prove-
´ ¸˜
detected by Tomaszewski et al. (1994). Using morphol- nientes de regiònes brasileiras produtoras de soja. Nematologia Bra-
ogy, isozyme profiles, karyology, host ranges, and RAPD sileira 27:1–12.
analysis, Carneiro et al. (1998) showed that the J2 iso- Cenis, J. L. 1993. Identification of four major Meloidogyne spp. by
random amplified polymorphic DNA (RAPD-PCR). Phytopathology
late did not exhibit any useful differentiating characters 83:76–80.
when compared with typical M. javanica isolates. Chacon, M. R., E. Rodriguez, R. M. E. Parkhouse, P. R. Burrows,
Meloidogyne cruciani, which occurred in French Gui- and T. Garate. 1994. The differentiation of parasitic nematodes using
10. 322 Journal of Nematology, Volume 37, No. 3, September 2005
random amplified polymorphic DNA. Journal of Helminthology 68: Janati, A., J. B. Berge, A. C. Triantaphyllou, and A. Dalmasso. 1982.
´
109–113. Nouvelles donne es sur l’utilisation des isoste rases pour
´ ´
Cliff, M. G., and H. Hirschmann. 1985. Evaluation of morphologi- l’identification des Meloidogyne. Revue de Nematologie 5:147–154.
´
cal variability in Meloidogyne arenaria. Journal of Nematology 17:445– Karssen, G. 1996. Description of Meloidogyne fallax n. sp. (Nema-
459. tode, Heteroderidade), a root-knot nematode from the Netherlands.
Cofcewicz, E. T., R. M. D. G. Carneiro, P. Castagnone-Sereno, and Fundamental and Applied Nematology 19:593–599.
P. Queneherve. 2004. Enzyme phenotypes and genetic diversity of
´ ´ ´ Karssen, G., T. Van Hoenselaar, B. Verkerk-Bakker, and R. Janssen.
root-knot nematodes parasitising Musa in Brazil. Nematology 6:85– 1995. Species identification of cyst and root-knot nematodes from
95. potato by electrophoresis of individual females. Electrophoresis 16:
Dalmaso, A., and J. B. Berge. 1978. Molecular polymorphism and
´ 105–109.
phylogenetic relationship in some Meloidogyne spp.: Application to
Pais, C. S., and I. M. O. Abrantes. 1989. Esterase and malate dehy-
the taxonomy of Meloidogyne. Journal of Nematology 10:323–332.
drogenase in Portuguese populations of Meloidogyne species. Journal
De Waele, D. 2000. Root-knot nematodes. Pp. 307–314 in D. R.
of Nematology 21:342–346.
Jones, ed. Diseases of banana, abaca, and enset. Wallingford, UK:
CAB International. Randig, O., M. Bongiovanni, R. M. D. G. Carneiro, and P. Castag-
Eisenback, J. D., and H. H. Triantaphyllou. 1991. Root-knot nema- none-Sereno. 2002. Genetic diversity of root-knot nematodes from
tode: Meloidogyne sp. and races. Pp. 191–274 in W. R. Nickle, ed. Brazil and development of SCAR markers specific for the coffee-
Manual of agricultural nematology. New York: Marcell-Dekker. damaging species. Genome 45:862–870.
Esbenshade, P. R., and A. C. Triantaphyllou. 1985. Use of enzyme Sambrook, J., E. F. Fritsch, and T. Maniatis. 1989. Molecular clon-
phenotypes for identification of Meloidogyne species. Journal of Nema- ing: A laboratory manual, 2nd ed. Cold Spring Harbor, NY: Cold
tology 17:6–20. Spring Harbor Laboratory Press.
Esbenshade, P. R., and A. C. Triantaphyllou. 1987. Enzymatic rela- Semblat, J. P., E. Wajnberg, A. Dalmasso, P. Abad, and P. Castag-
tionships and evolution in the genus Meloidogyne (Nematoda: Tylen- none-Sereno. 1998. High-resolution DNA fingerprinting of parthe-
chida). Journal of Nematology 19:8–18. nogenetic root-knot nematodes using AFLP analysis. Molecular Ecol-
Esbenshade, P. R., and A. C. Triantaphyllou. 1990. Isozyme pheno- ogy 7:119–125.
types for the identification of Meloidogyne species. Journal of Nema- Swofford, D. L. 1998. PAUP*. Phylogenetic Analysis Using UPGMA
tology 22:10–15. (*and Other Methods), vers. 4. Sinauer Associates, Sunderland, MA.
Fargette, M. 1987. Use of the esterase phenotype in the taxonomy Taylor, D. P., and C. Netscher. 1974. An improved technique for
of the genus Meloidogyne. 2. Esterase phenotype observed in West preparing perineal patterns of Meloidogyne spp. Nematologica 20:268–
African populations and their characterization. Revue de Nematolo-
´ 269.
gie 10:45–56.
Tomaszewski, E. K., M. A. M. Khalil, A. A. El-Deeb, T. O. Powers,
Felsenstein, J. 1985. Confidence limits on phylogenies: An ap-
and J. L. Starr. 1994. Meloidogyne javanica parasitic on peanut. Journal
proach using the bootstrap. Evolution 39:783–791.
of Nematology 26:436–441.
Garcia-Martinez, R., A. L. Taylor, and G. C. Smart, Jr. 1982. Meloido-
gyne cruciani n. sp., a root-knot nematode from St. Croix (U.S. Virgin Triantaphyllou, A. C. 1985. Cytological methods for study of oo-
Islands), with observations on morphology of this and two other spe- genesis and reproduction of root-knot nematode. Pp. 107–114 in K.
cies of the genus. Journal of Nematology 14:292–303. R. Barker, C. C. Carter, and J. N. Sasser, eds. An advanced treatise on
Gowen, S. R., and P. Queneherve. 1990. Nematode parasites of
´ ´ ´ Meloidogyne, vol. I, Methodology. Raleigh, NC: North Carolina State
bananas. Pp. 431–460 in M. Luc, R. A. Sikora, and J. Bridge, eds. University Graphics.
Plant-parasitic nematodes in subtropical and tropical agriculture. Welsh, J., and M. McClelland. 1990. Fingerprinting genomes using
Wallingford, UK: CAB International. PCR with arbitrary primers. Nucleic Acid Research 18:7213–7218.
Hirschmann, H. 1986. Meloidogyne hispanica n. sp. (Nematoda: Williams, G. K., A. R. Kubelick, K. J. Livak, J. A. Rafalski, and S. V.
Meloidogynidae), the Seville root-knot nematode. Journal of Nema- Tingey. 1990. DNA polymorphisms amplified with arbitrary primers
tology 18:520–532. are useful genetic markers. Nucleic Acid Research 18:6531–6535.