Ludivine Lassois

Crown Rot of Bananas: Preharvest Factors Involved in Postharvest Disease Development and Integrated Control Methods

Crown rot is a complex disease that affects export bananas in all banana-producing countries. Usually invisible when the fruits are packed for transportation from tropical countries to distant destinations, disease symptoms occur during shipment, ripening, and storage. This disease, characterized by rot and necrosis, affects tissues joining the fingers with each other, called the crown. It may reach the pedicel and even the banana pulp when crown rot is severe. Losses from 10 to 86% have been recorded for treated and untreated bananas, respectively. In this paper, we summarize the current knowledge on crown rot disease and associated control measures that must be considered throughout the production channel in order to be effective. We suggest a new approach to this postharvest disease of bananas: that of considering fruit quality potential in the field. This new concept of preharvest quality potential is a key factor to understanding crown rot development. Fruit quality potential depends on both a physio...

Identification of Genes Involved in the Response of Banana to Crown Rot Disease

Variations in banana susceptibility to crown rot disease have been observed but the molecular mechanisms underlying these quantitative host-pathogen relationships are still unknown. This study was designed to compare gene expression between crowns of banana fruit showing a high susceptibility (S(+)) and crowns showing a low susceptibility (S(-)) to the disease. Comparisons were performed at two situation times: i) between crowns (S(+) and S(-)) collected 1 h before inoculation and ii) between crowns (S+ and S-) collected 13 days after inoculation. Gene expression comparisons were performed with cDNA-amplified fragment length polymorphism (AFLP) and results were confirmed by real-time reverse-transcription polymerase chain reaction. Among genes identified as differentially expressed between S(+) and S(-) crowns, two were involved in signal transduction, three in proteolytic machinery, two had similarity to pathogenesis-related protein 14, one to a CCR4-associated factor protein, and one to a cellulose synthase. Paradoxically, the overexpression of the cellulose synthase gene was associated with banana showing a high susceptibility in both pre- and post-inoculation situations. Finally, the cDNA-AFLP identified a gene that seems to be associated with the quantitative banana responses to crown rot disease; this gene encodes a dopamine-β-monooxygenase, which is involved in the catecholamine pathway. To our knowledge, this work is the first to address both pre- and post-infection gene expression with the same host-pathogen combination and distinct susceptibility levels.

Dealing with paralogy in RADseq data: in silico detection and single nucleotide polymorphism validation in Robinia pseudoacacia L.

Abstract The RADseq technology allows researchers to efficiently develop thousands of polymorphic loci across multiple individuals with little or no prior information on the genome. However, many questions remain about the biases inherent to this technology. Notably, sequence misalignments arising from paralogy may affect the development of single nucleotide polymorphism (SNP) markers and the estimation of genetic diversity. We evaluated the impact of putative paralog loci on genetic diversity estimation during the development of SNPs from a RADseq dataset for the nonmodel tree species Robinia pseudoacacia L. We sequenced nine genotypes and analyzed the frequency of putative paralogous RAD loci as a function of both the depth of coverage and the mismatch threshold allowed between loci. Putative paralogy was detected in a very variable number of loci, from 1% to more than 20%, with the depth of coverage having a major influence on the result. Putative paralogy artificially increased the observed degree of polymorphism and resulting estimates of diversity. The choice of the depth of coverage also affected diversity estimation and SNP validation: A low threshold decreased the chances of detecting minor alleles while a high threshold increased allelic dropout. SNP validation was better for the low threshold (4×) than for the high threshold (18×) we tested. Using the strategy developed here, we were able to validate more than 80% of the SNPs tested by means of individual genotyping, resulting in a readily usable set of 330 SNPs, suitable for use in population genetics applications.

The susceptibility of bananas to crown rot disease is influenced by geographical and seasonal effects

Abstract Crown rot of banana fruits is caused by a complex of fungal pathogens, the most common of which is Colletotrichum musae, and is one of the main quality defects of exported bananas. Susceptibility of banana fruits to crown rot is influenced by many pre-harvest factors. The aim of this study was to improve on the methodology for the evaluation of fruit susceptibility and to verify whether cultivation areas in Cameroon as well as seasonal variations have an influence on the susceptibility to crown rot. Fruit susceptibility was evaluated on a monthly basis throughout a year (including the dry and rainy seasons) in three banana plantations located in very different agro-ecological conditions (two in a lowland area and one in a highland area). Fruit susceptibility was determined through an internal necrotic surface (INS) assessment after artificial inoculation with C. musae. The standardization of post-inoculation environmental conditions enabled more reliable INS assessments. Fruit susceptibility was found to be significantly influenced by cultivation area (P < 0.001) since fruits grown in low altitude (Dia-dia, Koumba, 80 m) were more susceptible than fruits grown in high altitude (Ekona, 500 m). Although no seasonal effect was observed (P = 0.075), there was a highly significant date effect (P < 0.001). This was specifically the case in low-altitude plantations where fruit susceptibility was higher for some harvest dates within the rainy season. In Ekona, fruit grade and number of leaves on the banana plant were found to be significantly higher than in the two other locations, while black leaf streak disease severity was significantly lower. The potential relationship with fruit susceptibility is fully discussed.

crown rot disease of bananas

Abstract Crown rot is a complex postharvest disease which affects export bananas in all banana-producing countries. A wide range of organisms are involved in crown rot of bananas but Colletotrichum musae is frequently considered as the most commonly isolated fungus and the most pathogenic one. Usually invisible when the fruits are packed for transportation from tropical countries to distant destinations, disease symptoms occur during shipment, ripening, and storage. This disease, characterized by rot and necrosis, affects tissues joining the fingers with each other, called the crown. It may reach the pedicel and even the banana pulp when crown rot is severe. Losses from 10% to 86% have been recorded for treated and untreated bananas, respectively. In most banana-growing areas, crown rot is principally controlled by postharvest fungicide treatments, but alternative control methods are being sought because of: (1) the emergence of resistance to some commonly used fungicides; (2) environmental problems linked to dumping of fungicide mixtures used at packing stations; and (3) consumer aversion to chemical residues in food. Important variations of both the severity of the damage and the nature of the complex are observed, depending on various pre- and postharvest factors. More specifically, the preharvest factors influence the parasitic and the physiological component of the fruit quality potential at harvest. The physiological component is defined as the sensitivity of the fruits to crown rot, and the parasitic component reflects the capacity of the parasitic complex to induce a level of disease. In this chapter, we summarize the current knowledge on crown rot disease and associated control measures which must be considered throughout the production channel in order to be effective as an integrated control strategy. The pre- and postharvest factors that favor infection are also discussed.

Thermotherapy, Chemotherapy, and Meristem Culture in Banana

Bananas that provide a staple food to the millions of people are adversely affected by several viruses such as Banana bunchy Top Virus (BBTV), Banana Streak Virus (BSV), and Cucumber Mosaic Virus (CMV). These viruses are known to have a devastating effect on crop production and constraint to the international exchange and conservation of banana germplasm-a cornerstone for breeding new cultivars. The viruses are particularly problematic in vegetative propagated crops, like bananas, because of their transmission in the planting material. Different virus eradication techniques have been developed, such as thermotherapy, chemotherapy, and meristem culture for providing virus-free planting material. Meristem culture proved to be the most effective procedure to eradicate phloem-associated viruses. This method requires isolation of meristematic dome of plant under the aseptic conditions and culture in an appropriate nutrient medium to develop new virus-free plants. Thermotherapy is another widely used virus eradication technique, which is initially carried out on in vivo or in vitro plants and eventually combined with meristem culture technique. The plantlets are initially grown at 28°C day temperature and increase it by 2°C per day until reaches 40°C and the night temperature at 28°C; maintain plants at 40°C for 4 weeks; excise meristem and culture onto the regeneration medium. In chemotherapy technique, antiviral chemical compound Virazole(®) is applied on meristem culture. Combination of these techniques is also applied to improve the eradication rate.

DNA taxonomy in the timber genus Milicia : evidence of unidirectional introgression in the West African contact zone

DNA-based techniques are helpful in characterising hybridisation patterns in plant species. To be efficient in disentangling species boundaries and interspecific gene flow, it is recommended to combine various methodologies and types of markers. Here, we used different analytical tools (algorithms implemented in Structure, Tess, NewHybrids and HIest, and the haploweb approach) and three nuclear genetic markers (7 nuclear simple sequence repeat loci (SSRs), 62 single-nucleotide polymorphism loci (SNPs) and a single-copy gene region, At103) to revisit hybridisation patterns in the commercially important African tree genus Milicia. Samples were collected in the natural ranges of Milicia regia and Milicia excelsa in West Africa. Using real data sets, simulated purebreds and hybrid genotypes, we found that SNPs yielded results more consistent than SSRs; outputs from the Bayesian and maximum-likelihood analyses differed significantly using the SSRs, whereas they were perfectly congruent using SNPs. A proportion of 12.4% hybrids were detected amongst the SNP genotype samples. A haploweb analysis of At103 gene sequences confirmed the existence of interspecific hybrids. There was also a clear evidence of advanced generations of hybrids (backcrossed individuals) but only towards M. regia. Although more investigation is required for understanding the mechanisms responsible for this asymmetric introgression, we suggest that it may be due to the differences in flowering time between species and between sexes, combined with a maternal inheritance of flowering time.

Lessons learned from the virus indexing of Musa germplasm: insights from a multiyear collaboration

The Bioversity International Transit Center (ITC) for banana hosts more than 1500 accessions largely covering the genetic diversity of the genus Musa. Its objective is to conserve this genetic diversity and to supply plant materials to users worldwide. All the Musa accessions must be tested for virus presence and, if infected, virus elimination must be attempted, to enable the supply of virus-free plant material. An international collaborative effort launched under the auspices of Bioversity International (2007–2013) finally led to the implementation of a two-step process to test the accessions. The first step, called pre-indexing, involved only molecular tests and was designed as a pre-screen of new germplasm lines or existing accessions to reduce the need for post-entry virus therapy and repeated virus indexing. The second step, called full indexing, was performed on either older existing accessions or newer accessions which tested negative during pre-indexing, and involved molecular tests, transmission electron microscopy (TEM) and symptom observation. In total, 270 germplasm lines (434 samples) were pre-indexed; while full indexing was carried out on 243 accessions (68 of which had been pre-indexed). A significant proportion of the samples tested during pre-indexing was infected with at least one virus (68%), showing the utility of this early pre-screening step. Banana streak OL virus and Banana mild mosaic virus were the most commonly detected viruses during both pre- and full indexing. For 22 accessions, viral particles were observed by TEM in full indexing while the molecular tests were negative, underlining the importance of combining various detection techniques. After full indexing, viruses were not detected in 166 accessions, which were then released for international distribution from the ITC. This publication exemplifies how the practical application of diagnostic protocols can raise fundamental questions related to their appropriate use in routine practice and the need for their continuous monitoring and improvement after their first publication.