The Prevalence and Distribution of Fusarium species in Norwegian Cereals: a Survey

In the period 1994–1996 a post-harvest survey was conducted in wheat, barley and oats to assess the occurrence and geographic distribution of Fusarium species in Norwegian cereals. The number of samples investigated was adjusted proportionally to the production of each cereal species within the regions. A total of 695 grain samples were analysed. The amount of Fusarium infection varied with cereal species and region of origin. The most frequently isolated Fusarium spp. from all samples were F. avenaceum, F. poae, F. tricinctum and F. culmorum. Other important toxigenic Fusarium spp. were F. graminearum, “powdery F. poae”, F. equiseti and F. sporotrichioides. A north-south gradient was valid for F. tricinctum, F. poae and in 1994 for “powdery F. poae”. In 1994 “powdery F. poae” was the most abundant potential producer of HT-2 and T-2 toxins in Norwegian cereals. Distribution of F. graminearum and F. culmorum demonstrated in this study, corresponded to previously reported DON-distribution, although DON seems to be produced by different species in different regions. Distribution of the isolated Fusarium species and comparison between cereals and locations are discussed.     

Impact of agronomic practices on populations of Fusarium and other fungi in cereal and noncereal crop residues on the Canadian Prairies

Fusarium head blight (FHB) is an important disease which has been causing damage to wheat and barley crops in western Canada. Because crop residues are an important source of inoculum, it is important to know the ability of Fusarium spp. to colonize and survive in different residue types, and how their populations might be affected by agronomic practices. Sampling of residue types on producers’ fields for quantification of Fusarium and other fungi was conducted in 2000–2001 in eastern Saskatchewan. Fusarium spp. were isolated from most fields, whereas their mean percentage isolation (MPI) was over 50% for cereal and pulse residues, and under 30% for oilseed residues. The most common Fusarium, F. avenaceum, had a higher MPI in pulse and flax (45–48%) than in cereal or canola (10–22%) residues. This was followed by F. equiseti, F. acuminatum, F. graminearum, F. culmorum and F. poae which were isolated from all, or most, residue types. Factors affecting Fusarium abundance in residues included the current crop, cropping history, and tillage system. In cereal residues, the MPI of F. avenaceum was higher when the current crop was another cereal (24%) versus a noncereal (4–8%). When the current crop was another cereal, the lowest MPI of F. avenaceum and F. culmorum occurred when the field had been in summerfallow (SF) two years previous (F. avenaceum: 17% for SF, 28% for a crop; F. culmorum: 1% for SF, 4% for a crop); in contrast, F. equiseti and Cochliobolus sativus were most common in residues of cereal crops preceded by SF (F. equiseti: 16% for SF, 10% for a crop; C. sativus: 22% for SF, 13% for a crop). The MPI of F. graminearum was higher when the crop two years previous was an oilseed (7%) versus a cereal (4%). In regards to tillage effects, when the current crop was a cereal, the MPI of F. avenaceum was higher under minimum (MT) and zero tillage (ZT) (22–37%) than conventional tillage (CT) (15%), that of F. graminearum was lowest under ZT (3% for ZT, 7–11% for CT-MT), whereas that of C. sativus was highest under CT (27% for CT, 6–11% for MT-ZT). Under ZT, previous glyphosate applications were correlated positively with F. avenaceum and negatively with F. equiseti and C. sativus. These observations generally agreed with results from previous FHB and root rot studies of wheat and barley in the same region. Percentage isolation of F. avenaceum from noncereal and of F. graminearum from cereal residues were positively correlated with FHB severity and percentage Fusarium-damaged kernels of barley and wheat caused by the same fungi.     

Application of random amplified polymorphic DNA to study genetic diversity in Paspalum scrobiculatum L. (Kodo millet, Poaceae)

Summary Genetic diversity and patterns of geographic variation among collections of Paspalum scrobiculatum (kodo millet) and P. polystachyum were studied using molecular markers generated through the random amplified polymorphic DNA (RAPD) method. A high level of polymorphism in RAPD markers was observed among the individual accessions, demonstrating the high genetic diversity of the crop. The markers obtained from the RAPD method were analyzed with the cluster analysis, principal coordinates and minimum spanning tree methods. Three major groups were resolved, one representing the African accessions, and two for the Indian accessions. The accessions of the north African kodo millet and P. polystachyum (considered conspecific with P. scrobiculatum) were quite distinct. The Australian kodo millet showed higher affinity to the African types. The study demonstrated that the RAPD technique can be applied to resolving degrees and patterns of genetic variation at the population and species levels, identifying cultivars, and defining gene pools of this crop.     

Funneliformis mosseae affects the root rot pathogen Fusarium oxysporum in soybeans

We analyzed the relationship between the dominant arbuscular mycorrhizal (AM) fungus Funneliformis mosseae and the dominant soybean root rot pathogen Fusarium oxysporum through the pot trials to help overcome obstacles to continuous cropping of soybean and to provide theoretical evidence that can be used to help prevent the reduced production induced by soybean root rot. Using qRT-PCR, we amplified the specific rDNA sequences of F. mosseae and F. oxysporum in soybean roots and rhizosphere soil and quantified the DNA contents of these fungi to determine the relationship between the dominant AM fungus F. mosseae and F. oxysporum. The DNA contents of F. oxysporum differed significantly depending on the presence of F. mosseae in both soybean roots and rhizosphere soil. Specifically, the DNA contents of F. oxysporum were reduced after inoculation with F. mosseae, suggesting that F. mosseae has a negative effect on the growth of F. oxysporum.     

Genetic diversity among varieties of Chia (Salvia hispanica L.)

Chia, Salvia hispanica L., was a staple crop in pre-Columbian Mesoamerica. Despite the great potential of the species as an oilseed crop, little research related to domesticated and wild varieties exists. A study was undertaken to assess genetic diversity among 38 wild and domesticated accessions of S. hispanica collected throughout Mesoamerica by using RAPD markers. Genetic diversity was higher among wild varieties (H _G= 0.15) than all domesticated varieties (H _G= 0.10) and modern commercial domesticated varieties (H _G= 0.02), suggesting a slight loss of diversity accompanying domestication and a near lack of diversity in modern commercial varieties. In addition, the preliminary results indicate that the center of genetic diversity is in the highlands of western Mexico.     

Differences in the apportionment of molecular and morphological variation in North American strawberry and the consequences for genetic resource management

Variation for 24 morphological traits measured in a greenhouse environment and 36 randomly amplified polymorphic DNA (RAPD) markers was assessed among 318 wild octoploid strawberry (Fragariaspp.) genotypes from diverse habitats across the northern USA. RAPD marker frequencies and certain leaf and flower morphology traits (petiole color, leaf mass/area ratio, leaflet length and width, flower and receptacle diameter, petal width, flowers/inflorescence) were significantly different between the F. chiloensis-platypetala and F. virginiana-glauca species complexes. The proportion of variation accounted for by provenance effects was lower for the RAPD markers than for most morphological traits, especially in the F. virginiana-glauca species complex. Morphological traits of potential adaptive importance group the collection into provenances within each species-complex, and reflect the significant habitat and geographic differences across the region from which the germplasm was collected. Variation among populations within provenances was low for the molecular and most morphological traits, with a much larger amount of variability among plants within populations. Most of the variation for the presumably more selectively-neutral RAPD data was among plants within populations and populations within provenances rather than among the provenances that were recognized based on morphological traits, especially in the F. virginiana-glauca complex. Patterns of diversity for morphological traits must be considered, along with more selectively-neutral molecular characters such as RAPDs, to formulate effective sampling strategies and to properly estimate the quantity and apportionment of diversity within this germplasm.     

Screening of culture conditions for pathogens of potato dry rot

Dry rot is a severe fungal disease during potato storage. Fusarium spp. is the pathogen causing severe rot of potato tubers, producing Fusarium toxins and contaminating food. The disease leads to a huge loss to China's potato industry. The present research optimized and screened the culture conditions of the major pathogens causing potato dry rot including Fusarium culmorum, Fusarium avenaceum, Fusarium sambucinum, Fusarium solani var. coeruleum, Fusarium trichothecioides, and Fusarium sporotrioides. The results showed that F. sambucinum and F. culmorum were sensitive to temperature and pH. F. sambucinum, F. trichothecioides, F. Solani var. coeruleum, and F. sporotrioides are favorable to the glucose-based culture while F. culmorum and F. avenaceum were to sucrose-based culture. Illumination stimulated the growth of F. sambucinum and F. sporotrioides but inhibited the growth of F. culmorum and F. avenaceum. It is concluded that the culture conditions are different with species of Fusarium and the understanding of the culture conditions is expected useful in research and the disease control.     

Identification and evaluation of <Emphasis Type="Italic">Forsythia</Emphasis> germplasm using molecular markers

This study identifies Forsythia germplasm and evaluated the genetic relationships of F. ×intermedia hybrids, cultivars and their putative parental species. Leaf samples of F. ×intermedia cultivars and species, such as F. koreana and F. suspensa, were collected in the Netherlands, Korea, and USA. Total genomic DNA was extracted and evaluated by randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) analyses. Dendrograms were constructed by using the neighbor-joining (NJ) clustering algorithm applying the interior branch (IB) test or analyzed by STRUCTURE. In the dendrogram generated by RAPD markers, two major clusters were observed. One cluster (CL-I) contained most of the F. ×intermedia cultivars, F. suspensa, and F. koreana. The other cluster (CL-II) included F. europaea, F. ovata, F. densiflora, F. mandshurica, F. japonica, F. viridissima, and cultivars derived from F. ovata. In the AFLP dendrogram, the placement of F. ×intermedia cultivars with F. suspensa was similar, forming cluster CL-A I. The RAPD and AFLP results clearly separated most F. ×intermedia cultivars from F. ovata derived cultivars. The full range of genetic diversity of F. suspensa and F. viridissima should be investigated to verify whether these two species are truly parental taxa for F. ×intermedia. Placement of F. viridissima, F. ovata, and F. japonica in different sub-clusters requires further investigation regarding genetic diversity in the species, and their close relationship with F. koreana, F. mandshurica, and F. saxatalis.     

Developing suppressive soil for root diseases of soybean with continuous long-term cropping of soybean in black soil of Northeast China

Field experiments were set up in 1991 to test if suppressive soil to soybean root diseases could be developed with continuous long-term soybean cropping in black soil of Northeast China. Based on the field observation in 2007, 2009, and 2011, soybean root growth was promoted, and the severities of root disease were reduced in a field with continuous long-term cropping with soybean. Population densities of the pathogens (Fusarium spp. and Heterodera glycines) in the soybean cropping field were significantly (P < 0.05) lower than rotation of soybean with wheat or corn. Higher levels of biological control agents (Trichoderma harzianum, Pochonia chlamydosporia, or Paecilomyces lilacinus) also were found in the long-term soybean cropping field. Therefore, continuous long-term cropping of soybean in black soil of Northeast China could develop suppressive soil to soybean root diseases.     

Geographic Pattern of Genetic Diversity Among 43 Ethiopian Mustard(Brassica carinata A. Braun) Accessions as Revealed by RAPD Analysis

As an oilseed crop, the cultivation of Ethiopian mustard (Brassica carinata) is restricted only to Ethiopia. Even though geographic diversity is a potent source of allelic diversity, the extent of genetic diversity among germplasm material of Ethiopian mustard from different countries has not been assessed. Forty-three accessions, comprising 29 accessions from eight different geographic regions of Ethiopia and 14 exotic accessions from Australia, Pakistan, Spain, and Zambia were analysed for their genetic diversity using random amplified polymorphic DNA (RAPD) technique. A set of 50 primers yielded a total of 275 polymorphic bands allowing an unequivocal separation of every Ethiopian mustard accession. The usefulness of the 50 RAPD primers in measuring heterozygousity and distinguishing accessions was variable such that polymorphic information content (PIC) varied from 0.05 to 0.40, band informativeness (BI) from 0.05 to 0.65 and primer resolving power (RP) from 0.15 to 6.83. Jaccard's similarity coefficients ranged from 0.44 to 0.87 indicating the presence of a high level of genetic diversity. On the average, Australian and Ethiopian accessions were the most similar while, Spanish and Zambian accessions were the most distant ones. Cluster analysis grouped the 43 accessions into four groups, which has quite a high fit (r = 0.80) to the original similarity matrix. With no prior molecular information, the RAPD technique detected large genetic diversity among the 43 accessions from five different countries and their grouping by dendrogram and principal coordinate analysis (PCoA) was inclined towards geographic differentiation of RAPD markers. Conversely, RAPD differentiation along geographic origin was not apparent within the Ethiopian accessions.     

A study of genetic diversity of sesame (<Emphasis Type="Italic">Sesamum indicum</Emphasis> L.) in Vietnam and Cambodia estimated by RAPD markers

Sesame (Sesamum indicum L.) is a traditional oil crop cultivated throughout South East Asia. To estimate the genetic diversity of this crop in parts at the region, 22 sesame accessions collected in Vietnam and Cambodia were analyzed using 10 RAPD markers. The 10 primers generated 107 amplification products of which 88 were polymorphic fragments (83%). Genetic diversity of all populations was H_t = 0.34 when estimated by Nei’s genetic diversity and species diversity was H′_sp = 0.513 when estimated by Shannon diversity index. Genetic distance ranged from 0.03 to 0.43, with a mean genetic distance of 0.23. The unweighted pair group method with arithmetic averages (UPGMA) cluster analysis for the 22 accessions divided the material in four groups. The dendrogram revealed a clear division among the sesame accessions based on their geographical region. Interestingly, some geographically distant accessions clustered in the same group, which might indicate the human factor involved in the spreading of sesame varieties. The high level of polymorphism shown suggests that RAPD techniques can also be useful for the selection of parents in sesame (Sesamum indicum L.) breeding program and for cultivar differentiation.     

新疆灰霉病菌多态性及其致病力分化分析

灰霉病菌(Botrytis cinerea)是引起植物病害的重要病原。本研究将来自新疆3个主要生态区域的12个灰霉病菌菌株, 经人工接种于7个寄主的离体叶片上测定病原菌的致病性, 并采用RAPD技术分析了12个菌株的遗传多态性。研究结果表明, 12个灰霉菌株的致病力存在明显差异, 可分为强、中、弱3种致病型, 其中来自和田的黄瓜灰霉菌和伊犁特克斯的番茄灰霉菌致病力最强; 阿克苏番茄灰霉菌的致病力最弱。根据在不同寄主上的致病反应, 12个菌株被划分为4个菌群。RAPD分析结果表明, 在0.65阈值下, 12个灰霉菌株可分为4个菌群, 说明供试灰霉菌株间有高度的遗传分化。灰霉病菌致病力强弱与菌株寄主、地理位置均无直接相关性, 但灰霉菌株的遗传距离与菌株来源存在明显相关性。     

Influence of Soil Moisture on Root Colonization of Glyphosate‐Treated Soybean by Fusarium Species

Abstract

The widespread use of glyphosate‐resistant (GR) cropping systems may impact rhizosphere microbial associations and crop productivity. It was previously reported that glyphosate accumulation in the rhizosphere may stimulate colonization of soybean [Glycine max (L.) Merr.] roots by soilborne Fusarium. Field studies often reveal inconsistent root colonization by Fusarium, especially during growing seasons characterized by contrasting rainfall patterns. Therefore, this study was conducted to determine the impact of different soil moisture contents on root colonization of glyphosate‐treated soybean by Fusarium species. Glyphosate (0.84 kg ae ha^?1) was applied to greenhouse‐grown glyphosate‐resistant (GR) soybean at the two to three trifoliate-leaf (V2–V3) growth stage growing in a Mexico silt loam at 27%, 13%, and 10% soil moisture contents. Soil and plant samples were sampled periodically after herbicide application and selectively cultured for Fusarium. Highest Fusarium colonization was associated with the glyphosate treatment, with maximum levels occurring at the highest soil moisture level. Thus, glyphosate interactions with root colonization by Fusarium in glyphosate‐resistant soybean are greatly influenced by soil moisture content.     

Soil microbial, fungal, and nematode responses to soil fumigation and cover crops under potato production

Sodium N-methyldithiocarbamate (metam sodium) and 1,3 dichloropropene are widely used in potato production for the control of soil-borne pathogens, weeds, and plant parasitic nematodes that reduce crop yield and quality. Soil fumigation with metam sodium has been shown in microcosm studies to significantly reduce soil microbial populations and important soil processes such as C and N mineralization. However, few published data report the impact of metam sodium on microbial populations and activities in potato production systems under field conditions. Fall-planted white mustard (Brassica hirta) and sudangrass (Sorghum sudanense) cover crops may serve as an alternative to soil fumigation. The effect of metam sodium and cover crops was determined on soil microbial populations, soil-borne pathogens (Verticillium dahliae, Pythium spp., and Fusarium spp.), free-living and plant-parasitic nematodes, and C and N mineralization potentials under potato production on five soil types in the Columbia Basin of Eastern Washington. Microbial biomass C was 8–23% greater in cover crop treatments compared to those fumigated with metam sodium among the soil types tested. Replacing fumigation with cover crops did not significantly affect C or N mineralization potentials. Cumulative N mineralized over a 49-day laboratory incubation averaged 18 mg NO₃-N kg⁻¹ soil across all soil types and treatments. There was a general trend for N mineralized from fumigated treatments to be lower than cover-cropped treatments. Soil fungal populations and free-living nematode levels were significantly lowered in fumigated field trials compared to cover-cropped treatments. Fumigation among the five soil types significantly reduced Pythium spp. by 97%, Fusarium spp. by 84%, and V. dahliae by 56% compared to the mustard cover crop treatment. The percentage of bacteria and fungi surviving fumigation was greater for fine- than coarse-textured soils, suggesting physical protection of organisms within the soil matrix or a reduced penetration and distribution of the fumigants. This suggests the potential need for a higher rate of fumigant to be used in fine-textured soils to obtain comparable reductions in soil-borne pathogens.     

Studying the extent of genetic diversity among Gossypium arboreum L. genotypes/cultivars using DNA fingerprinting

Genetic diversity is an area of concern for sustaining crop yield. Information on genetic relatedness/diversity among Gossypium arboreum L. cultivars/genotypes is scanty. We have used random amplified polymorphic DNA (RAPD) analysis to assess the genetic divergence/relationship among 30 genotypes/cultivars of G. arboreum. Of 45 primers surveyed, 63% were polymorphic. Out of the total number of loci amplified, 36% were polymorphic. The calculated genetic similarity between the cultivars/genotypes was in the range of 47.05–98.73%. Two genotypes, HK-244 and Entry-17, were the most distantly related. The average genetic relatedness among all the genotypes was 80.46%. However, most of the cultivated varieties showed a close genetic relationship, indicating a narrow genetic base in comparison to the non-cultivated germplasm. The calculated coefficients were used to construct a dendrogram using the unweighted pair group of arithmetic means (UPGMA) algorithm, which grouped the genotypes/cultivars into two major and three smaller clusters. The study is the first comprehensive analysis of the genetic diversity of G. arboreum germplasm and identifies cultivars that will be useful in extending the genetic diversity of cultivated varieties and future genome mapping projects.     

RAPD Polymorphism of the White-Flowered Gourd (Lagenaria siceraria (Molina) Standl. Landraces and its Wild Relatives in Kenya

The present study was performed to investigate genetic diversity of Kenyan landraces of the white-flowered gourd (Lagenaria siceraria), which exhibits tremendous morphological variation. RAPD analyses were performed on 53 landraces of the cultivated species L. siceraria and 42 accessions of three wild species (40 L. sphaerica, 1 L. abyssinica, and 1 L. breviflora). A total of 432 polymorphic bands were detected using 54 primers. The four species were clearly differentiated from one another. Intra-specific variations were investigated with L. siceraria and its wild relative L. sphaerica. Landraces of the cultivated species collected from different ethnic communities or regions were differentiated. Morphological variations were not associated with RAPD variations. Bitter landraces collected in Maasai communities showed two specific RAPD bands. In the wild species, accessions collected from the eastern and western sides of the Great Rift Valley were genetically differentiated from each other. In both species, genetic and geographical distance matrices computed among all pairs of accessions were significantly correlated, implying that the observed geographical variation can be explained by the 'Isolation by distance model'. Progeny plants derived from a common mother in L. siceraria showed a low level of segregation in RAPD pattern, suggesting that collected landraces are cultivated, maintaining their inherent traits although they are monoecious and insect-pollinated, whereas the wild relative L. sphaerica showed a higher level of segregation. The morphological diversity observed among landraces of L. siceraria is the result of human selection and their genetic identities are maintained by inbreeding probably resulting from frequent self-pollination.     

Distribution and biodiversity of soybean rhizobia in the soils of Shennongjia forest reserve,China

Soil samples were collected at an altitude of 500, 1,060, 1,500, 1,950, 2,400 and 3,100 m, respectively, from Shennongjia, a forest reserve in Hubei province (central China). Their corresponding pHs were 5.50, 4.91, 5.64, 5.28, 5.49 and 4.60. By using a plant trap method, a total of 25 soybean rhizobia were isolated from the soil above an altitude of 1,500 m and all identified to be Sinorhizobium fredii. Their genetic biodiversity was characterized by 16S–23S rDNA internally transcribed spacer (ITS) region polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) and random amplification DNA (RAPD) analysis. All the tested strains produced a 2.1 kb 16S–23S rDNA ITS fragment. After digestion with three restriction endonucleases (HaeIII, MspI and CfoI), respectively, great variations in 16S–23S rDNA ITS PCR-RFLP patterns were observed. The tested strains could be differentiated into 11 ITS genotypes. The genotypes of rhizobia were not related to geographical location. Twelve primers were applied to RAPD analysis and a dendrogram was obtained, showing that all the strains (including reference strain S. fredii USDA205) were divided into two diverging groups. Moreover, each group could be further divided into two subgroups. Both RAPD and 16S–23S rDNA ITS PCR-RFLP analysis indicated that a high degree of genetic diversity existed among S. fredii strains isolated from Shennongjia virgin soils. Since Shennongjia is an unexploited forest region in central China and the gene centre of soybean is located in China, the symbiotic genes harboured by these strains may be of great importance and the rich diversity of these strains might contribute to the adaptation of soybean to an alpine environment.     

Genetic variations and interrelationships in Vanilla planifolia and few related species as expressed by RAPD polymorphism

Vanilla is naturally distributed in Mexico and parts of Central America and the history of origin of cultivated vanilla suggests that the entire stock outside Mexico may be from a single genetic source. In the present study, RAPD polymorphism was used to estimate the level of genetic diversity and interrelationships among different collections of Vanilla planifolia Andr., and few related species, including both leafy and leafless types such as V. tahitensis J.W.Moore, V. andamanica, Rolfe, V. pilifera Holtt., and V. aphylla Blume. Studies revealed that there are very limited variation within collections of V. planifolia, indicative of its narrow genetic base, and of the related species we tested, V. tahitensis is nearest to V. planifolia. The species studied are diverse and have a similarity ranging from 1.2 to 57.3 %. Of the sampled taxa, V. andamanica is the most divergent and there is also reasonable variability within its collections, indicating the possibility of natural seed set. A total of 82 polymorphic bands expressed in the RAPD profiles were used to generate a genetic distance matrix, which was then used in cluster analysis. Specific groupings were revealed by the cluster analysis whereby the leafless forms (V. aphylla, V. pilifera and the new species) and V. andamanica formed separate clusters. This is the first report of species interrelationship studies, including both cultivated and wild vanilla species.     

Genetic variability of soybean bradyrhizobia populations under different soil managements

Due to their ecologic and economic importance, bradyrhizobia have been extensively studied in recent years. Since 1992, Bradyrhizobium elkanii SEMIA 587 and SEMIA 5019 and Bradyrhizobium japonicum SEMIA 5079 and SEMIA 5080 have been widely used in most Brazilian soybean fields. The objective of this work was to estimate the genetic variability of bradyrhizobial isolates recovered from soils under rhizobial inoculation and different soil managements. Only 25% of the isolates demonstrated high similarities to the original strains, and a strong correlation was obtained between the bradyrhizobial genetic variability and soil management. A high level of genetic diversity was observed both within isolates (H = 5.46) as well as among the different soil practices. Soil under no-tillage presented a higher bradyrhizobia diversity compared with bradyrhizobia isolated from soil under conventional tillage. Serological characterization also indicated that B. elkanii strains SEMIA 587 and SEMIA 5019 were more competitive and presented a higher nodular occupancy capacity than strains belonging to B. japonicum species in Southern Brazilian soils.