Congolese rhizospheric soils as a rich source of new plant growth-promoting endophytic Piriformospora isolates

In the last decade, there has been an increasing focus on the implementation of plant growth-promoting (PGP) organisms as a sustainable option to compensate for poor soil fertility conditions in developing countries. Trap systems were used in an effort to isolate PGP fungi from rhizospheric soil samples collected in the region around Kisangani in the Democratic Republic of Congo. With sudangrass as a host, a highly conducive environment was created for sebacinalean chlamydospore formation inside the plant roots resulting in a collection of 51 axenically cultured isolates of the elusive genus Piriformospora (recently transferred to the genus Serendipita). Based on morphological data, ISSR fingerprinting profiles and marker gene sequences, we propose that these isolates together with Piriformospora williamsii constitute a species complex designated Piriformospora (= Serendipita) 'williamsii.' A selection of isolates strongly promoted plant growth of in vitro inoculated Arabidopsis seedlings, which was evidenced by an increase in shoot fresh weight and a strong stimulation of lateral root formation. This isolate collection provides unprecedented opportunities for fundamental as well as translational research on the Serendipitaceae, a family of fungal endophytes in full expansion

Respiratory CO2 combined with a blend of volatiles emitted by endophytic Serendipita strains strongly stimulate growth of Arabidopsis implicating auxin and cytokinin signaling

Rhizospheric microorganisms can alter plant physiology and morphology in many different ways including through the emission of volatile organic compounds (VOCs). Here we demonstrate that VOCs from beneficial root endophytic Serendipita spp. are able to improve the performance of in vitro grown Arabidopsis seedlings, with an up to 9.3-fold increase in plant biomass. Additional changes in VOC-exposed plants comprised petiole elongation, epidermal cell and leaf area expansion, extension of the lateral root system, enhanced maximum quantum efficiency of photosystem II (Fv/Fm), and accumulation of high levels of anthocyanin. Notwithstanding that the magnitude of the effects was highly dependent on the test system and cultivation medium, the volatile blends of each of the examined strains, including the references S. indica and S. williamsii, exhibited comparable plant growth-promoting activities. By combining different approaches, we provide strong evidence that not only fungal respiratory CO2 accumulating in the headspace, but also other volatile compounds contribute to the observed plant responses. Volatile profiling identified methyl benzoate as the most abundant fungal VOC, released especially by Serendipita cultures that elicit plant growth promotion. However, under our experimental conditions, application of methyl benzoate as a sole volatile did not affect plant performance, suggesting that other compounds are involved or that the mixture of VOCs, rather than single molecules, accounts for the strong plant responses. Using Arabidopsis mutant and reporter lines in some of the major plant hormone signal transduction pathways further revealed the involvement of auxin and cytokinin signaling in Serendipita VOC-induced plant growth modulation. Although we are still far from translating the current knowledge into the implementation of Serendipita VOCs as biofertilizers and phytostimulants, volatile production is a novel mechanism by which sebacinoid fungi can trigger and control biological processes in plants, which might offer opportunities to address agricultural and environmental problems in the future

Present status of bacterial blight in cotton genotypes evaluated at Busia and Siaya counties of Western Kenya

Bacterial blight (BB) incited by Xanthomonas citri subsp. malvacearum (Xcm) is an important bacterial disease occurring in all cotton growing areas throughout the world, including parts of Western Kenya, that are characterized by a hot and humid climate. The disease causes seedling blight, angular leaf spot, boll rot and black arm on petioles and branches leading to a loss of fruiting branches with yield losses of up to 35 %. Fifty- one Kenyan cotton genotypes (Gossypium hirsutum L.) were established in the two counties of Siaya and Busia famous for their cotton cultivation in the western region. The BB symptoms caused by natural Xcm infection in the field were scored for each cotton accession. In addition artificial inoculation with the same strains was done to confirm the status under controlled greenhouse conditions. The results of BB disease scoring revealed that some accessions (e.g. T 24A and T 24B) possess a reasonable level of resistance. However, most of the Kenyan genotypes surveyed showed medium to severe symptoms of BB, including KSA 81 M, the only commercially grown cultivar in the region of Western Kenya. Overall, 71 % of the genotypes showed susceptibility and 29 % were classified as either resistant or moderately resistant. There is therefore need to improve the local commercial genotypes by introducing new genetic resources with a more durable BB resistance to ensure a successful revitalization of the Kenyan cotton sector

Glasshouse-specific occurrence of basal rot pathogens and the seasonal shift of Rhizoctonia solani anastomosis groups in lettuce

Basal rot is a common disease in Belgian lettuce, which is mainly controlled by fungicides and chemical soil disinfestation. A seasonal appearance of the basal rot pathogens: Rhizoctonia solani, Sclerotinia spp., Botrytis cinerea and Pythium spp. has been reported, but lettuce growers use standard spraying schemes, irrespective of the occurrence of the pathogen. Due to stricter regulations and environmental concerns the superfluous use of fungicides should be omitted. We investigated if the use of fungicides could be reduced by only controlling the active pathogens. Therefore, lettuce was continuously grown in three glasshouses without any fungal disease control and the active pathogens causing basal rot were identified. The occurrence of basal rot pathogens appeared to be glasshouse specific and the different basal rot pathogens were active throughout the year. However, a seasonal appearance of R. solani anastomosis groups and Pythium spp. was observed with AG4-HGI and Pythium ultimum active at higher temperatures and AG2-1, AG-BI, AG1-IB and Pythium sylvaticum at lower temperatures. We report for the first time the isolation of AG-BI from infected plants. Each R. solani anastomosis group had its own optimal growth rate in vitro. Differences in pathogenicity between R. solani anastomosis groups were observed on detached leaves. AG1-IB and AG4-HGI were most pathogenic, followed by AG2-1 and AG-BI. These results show that the fungicide spraying scheme should be adapted to the occurring pathogens in the glasshouse. This information is of high importance in developing a sustainable control strategy for basal rot pathogens

A thorough study of a Paratylenchus sp. in glasshouse-grown lettuce : characterisation, population dynamics, host plants and damage threshold as keys to its integrated management

In glasshouses practising monoculture of butterhead lettuce in Belgium, high densities of pin nematodes (Paratylenchusspp.) are frequently associated with reduced plant growth. Growers currently apply chemical soil disinfestation measures to manage this problem, although stricter phytosanitary regulations are forcing a shift towards integrated management. Efficient implementation of such management requires knowledge about the factors influencing nematode population dynamics, and the damage threshold for lettuce. The nematode populations in five Belgian glasshouses were monitored for at least 1 year by frequently soil sampling at 0-30 cm and 30-60 cm depth. An undescribed species of Paratylenchuswas identified in all glasshouses based on morphological and molecular features. High nematode densities (>20,000 (100 ml soil)(-1)) occurred in winter and spring. Chemical soil disinfestation lowered these populations greatly, although up to 14% survived in the deeper soil layer. After soil steaming under negative pressure, no pin nematodes were found. After 2 months of black fallow pin nematode densities were reduced by 50%-76%. Lamb's lettuce, parsley and wild rocket were found to be poor hosts in a pot experiment, while reproduction factors (P-f/P-i) on lettuce cultivars varied between 1 and 3. In three experiments with butterhead lettuce 'Cosmopolia' in pots with a series of 9 or 10 densities of Paratylenchussp. [up to 35,000 (100 ml soil)(-1)], no damage to lettuce heads was observed. However, root weight and root quality were reduced, and the corresponding damage thresholds were rather low [1,754 and 362Paratylenchussp. (100 ml soil)(-1), respectively]. Management strategies such as crop rotation, soil disinfestation or fallow are recommended to avoid pin nematode population build-up

Taxonomy and epidemiology of Pectobacterium and Dickeya spp. in Europe, North America and South Africa

<p>The blackleg-soft rot-aerial stem rot disease complex causes serious losses to the potato industry. It is caused by species of the genera <i>Pectobacterium</i> and <i>Dickeya</i>, collectively known as the soft rot Pectobacteriaceae. These soft rot Pectobacteriaceae also cause damage in a wide range of other host plants. <i>Pectobacterium brasiliense</i> has been the most prevalent in potato and pathogenic species in Europe and South Africa for the past decade, although the species composition is in constant flux due to the introduction of new species and taxonomic reclassification of current ones.</p><p>Updated information on the current species composition is required, as well as knowledge of possible differences in symptom expression between species. Such information would aid certification and diagnostic services in testing for the correct species and making accurate diagnoses.</p><p>Findings indicate that <i>Pectobacterium brasiliense</i> remains the most prevalent and widely distributed species in potato production areas. Other species that were identified included e.g. <i>Pectobacterium carotovorum</i>, <i>Pectobacterium parmentieri</i>, <i>Dickeya chrysanthemi</i> and <i>Pectobacterium versatile</i>. <i>Pectobacterium brasiliense</i> was also the most pathogenic species on potato. When looking at other host plants a wide variety of <i>Pectobacterium</i> and <i>Dickeya </i>species with large genetic variation occurs.</p><p>MALDI-TOF MS can only be used to identify <i>Pectobacterium</i> and <i>Dickeya </i>isolates at the genus level but preliminary results after improving the reference library look promising.</p><p>Given that there is also a large group of nonvirulent <i>P. brasiliense</i> isolates, a specific PCR which can differentiate between virulent and nonvirulent isolates of this species is being developed.</p&gt

Pratylenchus penetrans, a potential risk in glasshouse-grown lettuce : population dynamics and damage threshold

The root-lesion nematode, Pratylenchus penetrans, causes growth reduction in glasshouse-grown lettuce and is mainly controlled by chemical soil disinfestation. Integrated management strategies require more knowledge about the population dynamics and damage threshold densities. We monitored the population during 2.5 years in a commercial glasshouse by sampling soil in the same four 1 m(2) spots at 0-30 cm and 30-60 cm depth. The grower grew lettuce in rotation with leek, applied 1,3-dichloropropene in summer and left the field fallow during winter. Growing leek reduced the nematode population slightly but chemical soil disinfestation lowered the numbers drastically, although 41% of the nematodes in the deeper layer survived. Black fallow resulted in a slight increase of the population, probably due to hatching. Two pot experiments with ten densities of P. penetrans were conducted to estimate the damage threshold for a summer and autumn cultivar ('Cosmopolia' and 'Brighton', respectively). The thresholds for lettuce weight were 669 and 3834 P. penetrans (100 ml soil)(-1) in summer and autumn, respectively, but with considerable variability in estimated parameters. The thresholds for root damage were much lower: 204 and 48 P. penetrans (100 ml soil)(-1). Nematode numbers did not increase on lettuce in the pot tests (maximum multiplication rate was 0.40) but increased slightly in the commercial setting. These results show that populations of P. penetrans build up slowly when butterhead lettuce is rotated with leek and fallow, but chemical soil disinfestation is required to avoid numbers resulting in root damage

Fusarium isolates from Belgium causing wilt in lettuce show genetic and pathogenic diversity

Fusarium oxysporum f. sp. lactucae race 4 causes vascular necrosis and wilting of lettuce. First observed in Belgium in 2015, the lack of disease resistance in commercial cultivars allowed this pathogen to spread to nearly the entire Belgian production area within 4 years. Different levels of disease development were observed in different commercial greenhouses. To help explain this variation, we collected 78 Fusarium isolates and characterized them both physiologically and genetically. Molecular race identification indicated that 91% of the isolates belonged to race 4, while 6% of the isolates belonged to race 1, which was not previously reported in Belgium. Pathogenicity assays using differential cultivars confirmed the molecular race assignment of selected isolates. Cultivar Patriot was identified as a suitable new differential cultivar to race 4. Race 4 isolates were more aggressive than race 1 isolates at 24 degrees C, but only when using chlamydospore inoculum instead of a root dip assay containing microconidia. Variation in pathogenicity and aggressiveness of the races may explain differences in disease development in commercial greenhouses. Based on genotyping-by-sequencing (GBS), race 1 and race 4 isolates were highly similar to reference isolates. Fusarium curvatum, F. oxysporum f. sp. tulipae and F. oxysporum f. sp. rhois were phylogenetically separated from F. oxysporum f. sp. lactucae races 1 and 4 based on the GBS data, but not when using multilocus sequence data. Within F. oxysporum f. sp. lactucae race 4, the GBS data differentiated two rather homogeneous groups, suggesting at least two introductions. However, the two groups did not differ in aggressiveness