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Mosaic is the most common viral disease affecting fig plants. Although the Fig mosaic virus is the leading cause of mosaic disease, other viruses are also involved. High-throughput sequencing was used to assess viral infections in fig plants with mosaic. The genomic DNA and total RNAseq of mosaic-symptomatic fig leaves were sequenced using the Illumina platform. The analysis revealed the presence of fig badnavirus 1 (FBV-1), grapevine badnavirus 1 (GBV-1), citrus exocortis viroid (CEVd), and apple dimple fruit viroid (ADFVd). The FBV-1 and GBV-1 sequences were 7,140 bp and 7,239 bp long, respectively. The two genomes encode one open reading frame containing five major protein domains. The viroids, CEVd and ADFVd, were 397 bp and 305 bp long. Phylogenetic analyses revealed a close relationship between FBV-1 and Iranian isolates of the same species, while GBV-1 was closely related to Russian grapevine badnavirus isolates (Tem64, Blu17, KDH48, and Pal9). CEVd was closely related to other Iraqi isolates, while ADFVd was strongly related to a Spanish isolate. A registered endogenous pararetrovirus, caulimovirus-Fca1, with a size of 7,556 bp, was found in the RNA transcripts with a low expression level. This integrant was also detected in the genomes of the two lines 'Horaishi' (a female line) and 'Caprifig 6085' (a male line). Phylogenetic analyses revealed that caulimovirus-Fca1 was distinct from two other clades of different endogenous virus genera.

Background: The massive use of synthetic insecticides strongly affects the level of insecticide resistance in populations of Myzus persicae worldwide. The selection of target site insensitivity-mutations is particularly worrying in areas where agro-industrial crops are vulnerable to the attacks of aphids that vector viruses, as in the case of Tunisia. Knowledge of the resistance mechanisms evolved locally in this aphid pest is a prerequisite to improving and retaining the sustainability of integrated pest management strategies., Results: Target site mutations were surveyed in several populations of M. persicae collected from peach and potato crops between 2011 and 2017 in three Tunisian regions using real-time allele-specific PCR. The L1014F mutation (kdr locus) was found at a moderate frequency mostly in the heterozygous state and the homozygous resistant genotype was very uncommon. The M918T mutation (super-kdr locus) was present in a few heterozygous individuals, whereas the M918L mutation was detected for the first time in Tunisia and extreme North Africa. This latter mutation was shown to be widespread and well-established in Tunisia mainly as homozygous individuals, and was more abundant on peach than on potato crops. The S431F mutation (MACE) was found in a few heterozygous individuals. No individuals carrying the R81T mutation linked to neonicotinoid resistance were detected., Conclusion: This study points out a critical situation for the efficacy of pyrethroid insecticides to control M. persicae populations in Tunisia. It also confirms the rapid spread of the M918L mutation which has been detected in many different areas of the Mediterranean basin. © 2022 Society of Chemical Industry., (© 2022 Society of Chemical Industry.)

Here, we report on the morphological, molecular, and chemical characterization of a novel Fusarium species recovered from the roots and rhizosphere of Macrochloa tenacissima (halfa, esparto, or needle grass) in central Tunisia. Formally described here as F. spartum , this species is a member of the Fusarium redolens species complex but differs from the other two species within the complex, F. redolens and F. hostae , by its endophytic association with M. tenacissima and its genealogical exclusivity based on multilocus phylogenetic analyses. To assess their sexual reproductive mode, a polymerase chain reaction (PCR) assay was designed and used to screen the three strains of F. spartum , 51 of F. redolens , and 14 of F. hostae for mating type ( MAT ) idiomorph. Genetic architecture of the MAT locus in the former two species suggests that if they reproduce sexually, it is via obligate outcrossing. By comparison, results of the PCR assay indicated that 13/14 of the F. hostae strains possessed MAT1-1 and MAT1-2 idiomorphs and thus might be self-fertile or homothallic. However, when the F. hostae strains were selfed, 11 failed to produce perithecia and one only produced several small abortive perithecia. Cirrhi with ascospores, however, were only produced by 8/28 and 4/84 of the variable size perithecia, respectively, of F. hostae NRRL 29888 and 29890. The potential for the three F. redolens clade species to produce mycotoxins, pigments, and phytohormones was assessed by screening whole genome sequence data and by analyzing extracts on cracked maize kernel cultures via liquid chromatography-mass spectrometry.

The peach potato aphid, Myzus persicae (Sulzer), is a worldwide pest of many crops, and the most important aphid pest of peach and potato crops in Tunisia, mainly due to virus transmission, for which insecticides are frequently applied. We studied the genetic structure of M. persicae populations in Tunisia, in order to further our understanding of the biotic and abiotic factors shaping populations and to predict their evolutionary responses to the present management practices. We monitored peach orchards and seed potato crops in different seasons and regions from 2011-2013 and in 2016 (19 populations), assessing the genetic diversity of M. persicae at six microsatellite loci. Temporal and spatial changes in the frequency and distribution of 397 genotypes in 548 sampled aphids were studied. Only 37 genotypes were found more than once (clonal amplification), as most genotypes were found only once (91.60% in peach; 88.73% in potato crops). A similarly high genetic diversity was observed in aphids sampled from peach (G/N = 0.76; Ho = 0.617) and potato (G/N = 0.70; Ho = 0.641). Only a weak genetic differentiation among populations was found, mainly between geographic locations. Clustering analysis revealed genotypes to be grouped mainly according to host plant. The availability of the primary host, high proportion of unique genotypes, high genetic diversity and lack of structuring suggest that the aphid reproduces mainly through cyclical parthenogenesis in Tunisia. On the other hand, we provide a farm-scale study that shows how easily M. persicae can colonize different areas and hosts, which may have important implications in relation to plant virus vectoring., Competing Interests: The authors declare no conflicts of interest.

Prospecting of local grapevine ( Vitis vinifera L.) germplasm revealed that Tunisia possesses a rich patrimony which presents diversified organoleptic characteristics. However, viral diseases seriously affect all local grapevine cultivars which risk a complete extinction. Sanitation programs need to be established to preserve and exploit, as a gene pool, the Tunisian vineyards areas. The presence of the Grapevine leafroll associated virus-3 (GLRaV-3), Grapevine stem pitting associated virus (GRSPaV) and Grapevine virus A (GVA), were confirmed in a Tunisian grapevine cultivar using serological and molecular analyses. The association between GRSPaV and GVA viruses induces more rugose wood symptoms and damages. For this reason the cleansing of the infected cultivar is highly advisable. Direct and recurrent somatic embryos of cv. 'Hencha' were successfully induced from filament, when cultured on Chée and Pool (1987). based-medium, enriched with 2 mg 1 -1 of 2,4-dichlorophenoxyacetic acid and 2.5 mg 1 -1 of Thidiazuron, after 36 weeks of culture. After six months of acclimatization, RT-PCR carried on 50 somaplants confirmed the absence of GVA, GRSPa-V as well as GLRaV-3 viruses in all somaplants. Ampelographic analysis, based on eight OIV descriptors, was carried out on two years acclimated somaplants, compared to the mother plant. Results demonstrated that the shape and contours of 46 somaclones leaves are identical to mother plant leaves and four phenotypically off-type plants were observed. The healthy state of 100% 'Hencha' somaclones and the high percentage of phenotypically true-to-type plants demonstrate that somatic embryogenesis is a promising technique to adopt for grapevine viruses elimination.

Varroa destructor is one of the scourges of global beekeeping. It was detected for the first time in Benin in 2011 on the honeybee Apis mellifera adansonii. The aim of this study was to identify the strain of Varroa sp. found and study its genetic diversity. In total 183 Varroa mites were sampled in 21 municipalities in Benin. The COI intergenic region of each mite mtDNA was amplified by PCR. The SacI restriction enzyme was used to determine the strains of Varroa sp. Only the Korean (K) haplotype, identical to the most prevalent strain in Africa, was detected. Analysis of the genetic diversity of Varroa mites with eight microsatellite loci (Simple Sequence Repeats) indicated a very low diversity of genotypes. Thus, V. destructor populations from Benin appear to make up a single group. Their clonal wealth ranges from 0.00 to 0.47. This study is an important step forward in the monitoring of the infestation of V. destructor.

Background: We used molecular assays to diagnose resistance to pyrethroids and pirimicarb in samples of Myzus persicae from field crops or an insect suction trap in Tunisia. Genotypes for resistance loci were related to ones for polymorphic microsatellite loci in order to investigate breeding systems and patterns of genetic diversity, and to inform resistance management tactics., Results: The kdr mutation L1014F conferring pyrethroid resistance was found in all samples. The M918T s-kdr mutation also occurred in most samples, but only in conjunction with kdr. We discovered a previously unreported genotype heterozygous for L1014F but homozygous for M918T. Samples with modified acetylcholinesterase (MACE) conferring resistance to pirimicarb were less common but widespread. 16% of samples contained both the kdr and MACE mutations. Many unique microsatellite genotypes were found, suggesting that M. persicae is holocyclic in Tunisia. There were no consistent associations between resistance and microsatellite markers., Conclusion: This first study of insecticide resistance in M. persicae in North Africa showed genetic variation in insecticide resistance within microsatellite multilocus genotypes (MLG M s) and the same resistance mechanisms to be present in different MLG M s. This contrasts with variation in northern Europe where M. persicae is fully anholocyclic. Implications for selection and control strategies are discussed. © 2016 Society of Chemical Industry., (© 2016 Society of Chemical Industry.)

Our study is about the essential oil of Citrus aurantium L. in Tunisia and its plant extract. The yield of this essential oil is 0, 56% but the yield of the extract of plant was 17.1% for the aqueous extract ant 18.3% for the ethanolic extract. The analysis of chemical composition by using GC and GC/MS showed the essential oil of C. aurantium L. species to be rich in monoterpenes such as α-terpineol, lianolyl acetate, linalool and limonene. The antifungal activity of this oil showed us an inhibition of the germination of mushrooms, in the same way we could note that the biologic activities are generally assigned to the chemotypes high content in oxygenated monoterpene.

During a survey of legume crops in the northeast and northwest regions of Tunisia in April 2010, plants showing yellowing, reddening, and stunting symptoms were observed. A total of 281 symptomatic samples were collected: 142 plants from 10 chickpea (Cicer arietinum L.) fields, 84 plants from six faba bean (Vicia faba L.) fields, and 55 plants from six pea (Pisum sativum L.) fields. All samples were tested by the tissue-blot immunoassay procedure with the following monoclonal antibodies (MAbs): a broad-spectrum legume-luteovirus MAb (5G4), Faba bean necrotic yellows virus (FBNYV; genus Nanovirus, family Nanovirudae) (3-2E9; provided by J. Vetten, BBA, Braunschweig, Germany), Beet western yellows virus (BWYV; genus Polerovirus, family Luteoviridae) (A5977; Agdia, Elkhart, IN), Bean leafroll virus (BLRV; genus Luteovirus, family Luteoviridae) (4B10), Soybean dwarf virus (SbDV; genus Luteovirus, family Luteoviridae) (ATCC PVAS-650; American Type Culture Collection ATCC, Rockville, MD,), and a mixture of three MAbs (5-2B8, -3D5, and -5B8) to a Syrian isolate of Chickpea chlorotic stunt virus (CpCSV) (1). Serological tests showed that CpCSV was detected in 121 samples (43.06%) (62 chickpea, 57 faba bean, and 2 pea), followed by FBNYV (detected in three faba bean and three pea), BWYV (detected in three chickpea and one faba bean), and BLRV (detected in one pea sample). FBNYV, BLRV, and BWYV have been previously detected in faba bean and chickpea in Tunisia (4), but to our knowledge, this is the first report of CpCSV affecting legumes in Tunisia, which was found in seven chickpea, seven faba bean, and two pea fields. CpCSV has been reported to naturally infect legume crops such as chickpea, lentil, field pea, and faba bean as well as some leguminous weeds and a few wild non-legume plants species in many countries in West Asia and North Africa and causes economic losses on chickpea in Eritrea, Ethiopia, and Syria (1-3). Serological results of CpCSV was confirmed in four (two pea, one faba bean, and one chickpea) samples by reverse transcription (RT)-PCR using CpCSV specific primers (F:5'-TAGGCGTACTGTTCAGCGGG-3' and R:5'-TCCTTTGTCCATTCGAGGTGA-3') (3), which produced an amplicon of expected size (413 bp). No amplification was observed from healthy plant extracts. Sequence analysis revealed that the four Tunisian isolates (TuV 258-201 collected from faba bean [GenBank Accession No. HQ199310], TuC 215-201 collected from chickpea [HQ199307], and TuP 163-201 [HQ199308] and TuP 166-201 collected from pea [HQ199309]) were most similar to each other with a high sequence identity (99%) and clustered with isolates of CpCSV from Syria (GenBank Accession No. EU541270), Egypt (EU541269), and Morocco (EU541267), to which they were most closely related (98%). The Tunisian isolates also showed high sequence identity (96%) in the coat protein region with Ethiopian (GenBank Accession No. EU541257) and Sudanese (EU541263) isolates. However, all isolates are distinct from BWYV, BLRV, and SbDV (less than 70% sequence identity). Since CpCSV is transmitted by aphids only, additional studies are needed to identify the host range of the virus and the efficient aphid vectors to better understand the epidemiology of this virus under Tunisian conditions References: (1) A. D. Abraham et al. Arch.Virol. 154:791; 2009. (2) N. Y. Asaad et al. J. Phytopathol. 157:756, 2009. (3) S. G. Kumari et al. Phytopathol. Mediterr. 47:42, 2008. (4) A. Najar et al. Phytopathol. Mediterr. 39:423, 2000.

Fusarium culmorum is a major pathogen associated with Fusarium head blight (FHB) of wheat in Tunisia. It may cause yield loss or produce mycotoxins in the grain. The objectives of the present study were threefold: to evaluate by PCR assays the type of mycotoxins produced by 100 F. culmorum isolates recovered from different regions in Northern Tunisia, to determine the amount of mycotoxin production by HPLC analysis, and to analyse for correlations between the amount of mycotoxin produced and the aggressiveness of isolates. PCR assays of Tri5, Tri7, Tri13, and Tri3 were used to predict whether these isolates could produce nivalenol, 3-acetyl-deoxynivalenol, or 15-acetyl-deoxynivalenol. Two of the isolates were predicted to produce NIV, whereas the others were predicted to produce 3-AcDON. Trichothecene production was confirmed and quantified by high pressure liquid chromatography (HPLC) in 28 isolates, after growth on wheat grains, and in a liquid Mycotoxin Synthetic medium (MS). All strains produced DON/3-AcDON at detectable levels ranging from 21 microg/g to 11.000 microg/g of dry biomass on MS medium and from 10 microg/g to 610 microg/g on wheat grain. The evaluation of the relationship between 3-AcDON production and aggressiveness of 17 strains revealed a significant difference in aggressiveness among the isolates. Moreover, only a significant correlation was revealed between aggressiveness and the amount of 3-AcDON produced on MS medium (r=0.36). Chemotyping of F. culmorum isolates is reported for the first time for isolates from Tunisia, and highlights the important potential of F. culmorum to contaminate wheat with 3-AcDON trichothecenes., ((c) 2010 Elsevier B.V. All rights reserved.)

Grapevine leafroll associated virus-3 (GLRaV-3) and Grapevine leafroll associated virus-5 (GLRaV-5), two members of the genus Ampelovirus associated with grapevine leafroll disease, were transmitted by the mealybug Planococcus ficus and the soft scale insect Ceroplastes rusci from infected to healthy vines under experimental conditions. The efficiencies of transmission of GLRaV-3 and GLRaV-5 by P. ficus were 23.3 and 8.3%, respectively, and by C. rusci were 3.3 and 1.7%, respectively. Juvenile instars of P. ficus were more efficient in transmission of the viruses than adult females. This is the first report of the ability of C. rusci to transmit these viruses to grapevines.

Surveys of yellowing viruses under nonheated and geothermal heated plastic tunnels and in open field crops of melon (Cucumis melo), cucumber (C. sativus), zucchini (Cucurbita pepo), squash (C. maxima), watermelon (Citrullus lanatus), and ware cucurbit (Ecballium elaterium) were carried out year-round during 2000-2001, 2003, and 2004 in the major cucurbit-growing areas in Tunisia. Severe yellowing symptoms on older leaves of cucurbits were observed in open fields and under plastic-tunnel production systems. These yellowing symptoms and large populations of aphids (Aphis gossypii) on a diversity of cucurbit crops in Tunisia support the hypothesis of a viral cause of the disease. Virus identification using double-antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA), followed by reverse transcription-polymerase chain reaction (RT-PCR) and immunocapture (IC)-RT-PCR showed that Cucurbit aphid-borne yellows virus (CABYV) was largely distributed in melon, cucumber, zucchini, squash, and watermelon crops. Ware cucurbit (E. elaterium) and lettuce (Lactuca sativa) crops were identified as potential CABYV reservoirs. The RT-PCR-amplified partial coat protein (CP) and P4 genes were cloned and sequenced from nine Tunisian CABYV isolates. CP and P4 gene nucleotide and amino acid sequence comparisons as well as phylogenetic reconstructions showed that the Tunisian isolates clustered into two major subgroups. Comparisons with CABYV sequences retrieved from GenBank showed high nucleotide and CP amino acid identities, and close relationships of the Tunisian isolates with Italian and French isolates.

Grapevine leafroll disease is one of the most important diseases that occurs in cultivated grapevines in the world. So far, nine serologically distinct viruses of the family Closteroviridae have been isolated from diseased vines (3). A previous study (4) has shown that Grapevine leafroll-associated viruses (GLRaV) -1, -2, and -3 are present in Tunisian grapevines and GLRaV-3 is the predominant virus associated with leafroll disease. A survey was conducted in table grapes to identify other viruses associated with this disease. Samples of dormant canes were collected and screened by indirect Biotin Steptavidin ELISA with specific antibodies to GLRaV-5 (Bio-Rad, Sanofi, France) according to the manufacturer's instructions. Serological analysis revealed that nearly 47% of the samples were infected with GLRaV-5. To confirm GLRaV-5 identification and identify other leafroll viruses, vines with severe leafroll symptoms were collected and total RNA extracts were obtained from six samples and tested at Waite Diagnostics (University of Adelaide, Australia) by reverse transcription (RT)-PCR using primers for GLRaV-5 (2), LR5-1F 5'-CCCGTGATACAAGGTAGGACA-3' and LR5-1R 5'-CAGACTTCACCTCCTGTTAC-3' with a resulting amplicon size of 690 bp and primers for GLRaV-9, LR9F 5'-ACAGTGGTCGGCATAAGAAAAG-3' and LR9R 5'-ACACAAACATGCAGGCCAAAG-3' with a resulting amplicon size of 250 bp. Results showed that 1 of 6 and 5 of 6 of the samples were infected with GLRaV-5 and GLRaV-9, respectively, by RT-PCR and comparable results were obtained by ELISA. Amplicons were cloned and sequenced to confirm the identification of GLRaV-5 and GLRaV-9. The obtained sequences showed 99.1% nt identity and 94.8% amino acid similarity with an isolate of GLRaV-5 (GenBank Accession No. AF233934) and 97.6% nt identity and 94.8% amino acid similarity with an isolate of GLRaV-9 (GenBank Accession No. AY297819). The occurrence of GLRaV-9 has previously been reported in California and Australia (1). To our knowledge, this is the first report on the occurrence of GLRaV-5 and -9 in Tunisian grapevines. The widespread occurrence of GLRaV-5 and -9 is probably due either to the presence of their putative vectors, Planococcus ficus (Signoret) and Planococcus citri (Risso), or by propagation using infected local source material. Further studies are in progress to verify the implication of indigenous mealybugs in the spread of these viruses. References: (1) R. Alkowni et al. J. Plant Pathol. 86:123, 2004. (2) X. Good and J. Monis. Phytopathology 91:247, 2001. (3) P. Gugerli. ICVG, Extended Abstracts 14:23, 2003. (4) N. Mahfoudhi et al. EPPO Bulletin 28:197, 1998.

Viruses, distributed worldwide on cucurbits, cause severe damage to crops. Virus surveys in 2003 and 2004 were made in all the major cucurbit-growing areas in Tunisia. Large populations of aphids (Aphis gossypii Glover) and severe yellowing symptoms of older leaves of cucurbits were observed in outdoor and under plastic-tunnel cultivation, suggesting the presence of Cucurbit aphid-borne yellows virus (CABYV, genus Polerovirus, family Luteoviridae). Leaf samples collected from symptomatic and asymptomatic plants of melon (Cucumis melo L.), cucumber (C. sativus L.), squash (Cucurbita pepo L.), watermelon (Citrullus lanatus L.), and ware cucurbit (Ecballium elaterium L. T. Richard) were screened for the presence of CABYV using enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). Reference isolate, CABYV-N (GenBank Accession No. X76931) was provided by H. Lecoq (INRA-Monfavet Cedex, France). Sample extracts from fresh leaf tissues were tested using ELISA with an antiserum prepared against this isolate. In addition, total RNA was extracted from fresh leaf tissues according to the technique of Celix et al. (2) using the Titan RT-PCR kit from Roche Diagnostics (Penzberg, Germany). Forward primer (5'-GAGGCGAAGGCGAAGAAATC-3') and reverse primer (5'-TCTGGACCTGGCACTTGATG-3') were designed with the available sequence of the reference isolate. ELISA tests demonstrated that 91 plants were positive among 160 plants tested with severe yellowing symptoms. All asymptomatic plants were negative. RT-PCR results yielded an expected 550-bp product that was amplified from the reference isolate. Of the 160 plants tested using ELISA, 106 plants were screened with RT-PCR including the 91 plants that were positive in ELISA. These 91 plants also were positive after RT-PCR amplification as were 12 more plants. This demonstrated that the RT-PCR test is more sensitive. No amplicons were produced from extracts of asymptomatic plants, RNA preparations of Cucurbit yellow stunting disorder virus (CYSDV), or Beet pseudo yellows virus (BPYV) positive controls provided by B. Falk (University of California, Davis). CYSDV and BPYV can induce similar yellowing symptoms in cucurbits. The results of the ELISA and RT-PCR tests showed that CABYV is widely distributed on five cucurbit species in the major growing areas of Tunisia including the northern, Sahel, central, and southern regions where it was detected, respectively, in 10 of 25, 11 of 21, 24 of 37, and 58 of 77 samples tested. CABYV was detected at the rates of 63 of 72 on melon, 10 of 21 on cucumber, 17 of 24 on squash, 10 of 25 on watermelon, and 3 of 18 on ware cucurbit. CABYV also seems to be widespread throughout the Mediterranean Basin (1,3,4), but to our knowledge, this is the first report of the occurrence of CABYV in Tunisia on different species of cucurbit and ware cucurbit. References: (1) Y. Abou-Jawdah et al. Crop Prot. 19:217, 2000. (2) A. Celix et al. Phytopathology 86:1370, 1996. (3) M. Juarez et al. Plant Dis. 88:907, 2004. (4) H. Lecoq et al. Plant Pathol. 41:749, 1992.