Outlook of Cassava Brown Streak Disease Assessment: Perspectives of the Screening Methods of Breeders and Pathologists

Cassava production and productivity in Eastern, Central, and Southern Africa are ravaged by cassava brown streak disease (CBSD), causing yield losses of up to 100% when susceptible varieties are grown. Efforts to develop CBSD-resistant clones are underway. However, the methods for screening CBSD resistance currently vary between breeders and pathologists, with the limited empirical data to support their choices. In this study, we used the empirical CBSD foliar and root necrosis data from two breeding populations, termed cycle zero (C0) and cycle one (C1), to assess and compare the effectiveness of the CBSD screening methods of breeders vs. pathologists. On the one hand, the estimates of broad-sense heritability (H2) for the CBSD root necrosis assessment of breeder ranged from 0.15 to 0.87, while for the assessment method of pathologists, H2 varied from 0.00 to 0.71 in C0 clones. On the other hand, the marker-based heritability estimates (h2) for C0 ranged from 0.00 to 0.70 for the assessment method of breeders and from 0.00 to 0.63 for the assessment method of pathologists. For cycle one (C1) population, where both foliar and root necrosis data were analyzed for clones assessed at clonal evaluation trials (CETs) and advanced yield trials (AYTs), H2 varied from 0.10 to 0.59 for the assessment method of breeders, while the H2 values ranged from 0.09 to 0.35 for the CBSD computation method of pathologists. In general, higher correlations were recorded for foliar severity from the assessment method of breeders (r = 0.4, p ≤ 0.01 for CBSD3s and r = 0.37, p ≤ 0.01 for CBSD6s) in C1 clones evaluated at both clonal and advanced breeding stages than from the approach of pathologists. Ranking of top 10 C1 clones by their indexed best linear unbiased predictors (BLUPs) for CBSD foliar and root necrosis showed four overlapping clones between clonal and advanced selection stages for the method of breeders; meanwhile, only a clone featured in both clonal and advanced selection stages from the CBSD assessment method of pathologists. Overall, the CBSD assessment method of breeders was more effective than the assessment method of pathologists, and thus, it justifies its continued use in CBSD resistance breeding.

Pythium spp. associated with root rot and stunting of winter crops in North Carolina

Annual double-crop rotation systems that incorporate winter wheat, clary sage, or a cover crop are common in eastern North Carolina. Stunting and root rot of clary sage (Salvia sclarea L.) reduce yields of this crop, especially in wet soils. Stunting and reduced stand establishment also afflict winter cover crops, including rye, rapeseed, and winter pea. Pythium spp. are causal agents of root rot of winter wheat in this region, but their role in root rot and stunting of other winter crops is not understood. During the growing seasons of 2018-2019 and 2019-2020 samples of clary sage, rye, rapeseed, and winter pea displaying symptoms of stunting were collected across eastern NC, resulting in the recovery of 420 isolates of Pythium from the roots of all hosts. P. irregulare, P. spinosum, and the complex Pythium sp. cluster B2A were the most frequently isolated species from clary sage. P. irregulare and P. spinosum were aggressive pathogens of clary sage at 18°C, and caused moderate root rot at 28°C. Koch’s postulates confirmed that isolates belonging to Pythium sp. cluster B2A, P. sylvaticum, P. pachycaule, P aphanidermatum, P. myriotylum, and P. oopapillum are pathogens of clary sage. P. irregulare (37% of all isolates) and members of the species complex Pythium sp. cluster B2A (28% of all isolates) comprised the majority of isolates collected from all hosts and were the most frequently isolated species from rye, rapeseed, and winter pea. In pathogenicity assays, isolates representing P. irregulare and P. spinosum caused slight to moderate root necrosis on rye, rapeseed, and winter pea. Isolates representing Pythium sp. cluster B2A caused slight to moderate root necrosis on rapeseed and clary sage, but no symptoms on rye or winter pea.

Evaluation of the phytotoxicity of 2,4-diacetylphloroglucinol and Pseudomonas brassicacearum Q8r1-96 on different wheat cultivars

Pseudomonas brassicacearum Q8r1-96 and other 2,4-diacetylphloroglucinol (DAPG)-producing pseudomonads of the Pseudomonas fluorescens complex possess both biocontrol and growth-promoting properties and play an important role in suppression of take-all of wheat in the Pacific Northwest (PNW) of the United States. However, P. brassicacearum can also reduce seed germination and cause root necrosis on some wheat cultivars. We evaluated the effect of Q8r1-96 and DAPG on the germination of 69 wheat cultivars that have been or currently are grown in the PNW. Cultivars varied widely in their ability to tolerate P. brassicacearum or DAPG. The frequency of germination of the cultivars ranged from 0 to 0.87 and from 0.47 to 0.90 when treated with Q8r1-96 and DAPG, respectively. There was a significant positive correlation between the frequency of germination of cultivars treated with Q8r1-96 in assays conducted in vitro and in the greenhouse. The correlation was greater for spring than for winter cultivars. In contrast, the effect of Q8r1-96 on seed germination was not correlated with that of DAPG alone, suggesting that DAPG is not the only factor responsible for the phytotoxicity of Q8r1-96. Three wheat cultivars with the greatest tolerance and three cultivars with the least tolerance to Q8r1-96 were tested for their ability to support root colonization by strain Q8r1-96. Cultivars with the greatest tolerance supported significantly greater populations of strain Q8r1-96 than those with the least tolerance to the bacteria. Our results show that wheat cultivars differ widely in their interaction with P. brassicacearum and the biocontrol antibiotic DAPG.

Evaluation of Newly Released Cassava Varieties for Yield Performance, Reactions to Cassava Diseases and Farmers’ Preference in Adjumani District of Uganda

Cassava viral diseases particularly cassava brown streak disease (CBSD) and cassava mosaic disease (CMD) have put pressure on cassava breeders to develop varieties that are resistant/tolerant to them. Several cassava varieties have been rolled out to farmers with the latest being NAROCASS series that are tolerant to these diseases. The yield performance of these new varieties have not been documented in some sub zones like Adjumani district that falls within a major West Nile agro-ecology of Uganda. Therefore this study sought to established yield performances of, reactions to major diseases, and farmers’ preference to these newly released cassava varieties in Adjumani. Results showed significant (P ≤ 0.05) differences among cassava varieties and experimental sites for all the parameters evaluated. Average yield performance by varieties were in the order of a local cassava—Alifasia (8.7 t/Ha) lowest, NAROCASS 2 (18.55 t/Ha), NASE 14 (33.97 t/Ha), NASE 19 (41.26 t/Ha), and NAROCASS 1 (41.71 t/Ha) highest. CMD foliar symptom was present at all sites on a local cassava—Alifasia, and on NAROCASS 1 in Ayiri parish, Ukusijoni sub-county. CBSD foliar symptoms were observed on off-types (TME 14) in the plot of NASE 14 in Miniki Parish only whereas CBSD root necrosis was observed at all sites on the local cassava—Alifasia, and on NASE 19 in Maaji parish, Ukusijoni sub-county. Cassava root rot disease was localised in Ukusijoni sub-county only. Farmers’ preferences to these newly released cassava varieties were in the order of NASE 19 (40.96%), NAROCASS 1 (24.86%), NAROCASS 2 (15.82.28%), NASE 14 (15.54%), and a Local cassava—Alifasia (2.83%). Result from this study strengthens the information gap in the breeding process towards developing a cassava variety with farmer-preferred attributes, and can also inform the utilisation of these improved cassava varieties in Adjumani district.

RNA silencing machinery contributes to inability of BSBV to establish infection in Nicotiana benthamiana: evidence from characterization of agroinfectious clones of Beet soil-borne virus

Beet soil-borne virus (BSBV) is a sugar beet pomovirus frequently associated with Beet necrotic yellow veins virus, the causal agent of the rhizomania disease. BSBV has been detected in most of the major beet-growing regions worldwide, yet its impact on this crop remains unclear. With the aim to understand the life cycle of this virus and clarify its putative pathogenicity, agroinfectious clones have been engineered for each segment of its tripartite genome. The biological properties of these clones were then studied on different plant species. Local infection was obtained on agroinfiltrated leaves of Beta macrocarpa. On leaves of Nicotiana benthamiana, similar results were obtained, but only when heterologous viral suppressors of RNA silencing were co-expressed or in a transgenic line down regulated for both dicer-like protein 2 and 4. On sugar beet, local infection following agroinoculation was obtained on cotyledons, but not on other tested plant parts. Nevertheless, leaf symptoms were observed on this host via sap inoculation. Likewise, roots were efficiently mechanically infected, highlighting low frequency of root necrosis and constriction, and enabling the demonstration of transmission by the vector Polymyxa betae. Altogether, the entire viral cycle was reproduced, validating the constructed agroclones as efficient inoculation tools, paving the way for further studies on BSBV and its related pathosystem.

FaRCa1 Confers Moderate Resistance to the Root Necrosis Form of Strawberry Anthracnose Caused by Colletotrichum acutatum

FaRCa1 is a major locus conferring resistance to anthracnose fruit rot (AFR) caused by Colletotrichum acutatum, an important pathogen of strawberry (Fragaria ×ananassa). The objective of this study was to characterize the effects of FaRCa1 on anthracnose root necrosis (ARN) via root inoculations and DNA marker characterization of the locus. A subgenome-specific high-resolution melting (HRM) marker for an insertion/deletion (InDel) near FaRCa1 was designed using the ‘Camarosa’ octoploid reference genome. The marker was used to genotype cultivars and advanced selections studied in two seasons. A root disease screening method was developed in which roots were cut and dipped in a spore suspension before planting, using a mixture of three local isolates of the C. acutatum species complex. ARN was indirectly scored by calculating image-based leaf area differences among inoculated and noninoculated plants. The allele of FaRCa1 conferring resistance to AFR also conferred a significant reduction in ARN. Thus, a robust and easily scored DNA test is now available to breeders for selecting for resistance to both the fruit and root forms of strawberry anthracnose.

Optimum Time for Harvesting Cassava Tubers to Reduce Losses Due to Cassava Brown Streak Disease in Northeastern DRC

The present study aimed to determine the appropriate time to harvest cassava tuberous root which minimize the losses due to cassava brown streak disease (CBSD) in the region of Yangambi, DRC. To achieve the aim of the study, 38 cassava cultivars were evaluated in Yangambi INERA’s Research Center for CBSD in the roots at harvest time between 9 and 13 months after planting (MAP). All the 38 cultivars tested showed CBSD root necrosis symptoms. Foliar symptoms occurred on 37.6% of the evaluated cultivars while CBSD root necrosis varied significantly among cultivars (7.0% to 82.5%) depending on susceptibility and the age of plant. This indicates the differential response of the cultivars to CBSD infection. Whitefly population density decreased with age of cassava, it was of 3 whiteflies per plant (9 MAP) to 1 (10 MAP). We noticed that in older plants, whitefly population decreased from 1 at 11 MAP to none at 13 MAP. Although, some cultivars did not show CBSD symptoms up to 12 MAP, they were not necessarily less attractive to whitefly. Negative relationship (r = -0.08 and r = -0.25) has been found between whitefly number and foliar symptom severity or between whitefly and tuber necrosis severity. Beyond 12 MAP, CBSD necrosis (severity score 4) was present in the tubers of 3 cultivars (EUR/2011/0148, Yafelamonene and Ybi/2011/258). Our study shows that in order to mitigate the losses due to CBSD necrosis, the optimum harvesting time for cassava tubers in Yangambi is 9 MAP.

Cassava Root Necrosis Disease (CRND): A New Crop Disease Spreading in Western Democratic Republic of Congo and in Some Central African Countries

Cassava is consumed in the Democratic Republic of Congo (DRC) as a staple food for the majority of the Congolese population. This crop is used in several forms: as fufu, chikwangue and pondu; cassava leaves are the most consumed vegetable in the country. In 2002, cassava root symptoms similar to cassava brown streak disease (CBSD) were reported for the first time in western DRC. PCR assays, using primers specific to Cassava brown streak virus (CBSV), failed to detect or identify any viral pathogens in diseased cassava samples from western DRC. Therefore, next generation sequencing (NGS) techniques were used as they are able to sequence full organism genomes and are widely used for the identification of pathogens responsible for new diseases. The main objective of this study was to identify the pathogens causing root necrosis in western DRC. Whatman®FTA™ cards were used to collect 12 cassava leaf samples from plants with symptoms indicative of very severe root necrosis, as well as two asymptomatic samples. These 12 samples were sent to Australia at the University of Western Australia in Perth for next generation sequencing (NGS) using the Illumina HiSeq platform. Additional bioinformatics tools included Geneious, CLC workbench, ParaKraken and Kaijou software for short DNA sequences. No viruses (including CBSV) were found in any of the DRC samples. These preliminary results confirm all the previous negative results obtained using PCR and CBSV primers. However, NGS analyses did reveal the presence of a number of bacterial and fungal taxa. These will require further investigation and tests such as the Koch Postulates, to establish their specific pathogenic role in cassava. This is the first scientific evidence that no currently known virus is responsible for the disease which had been referred to previously as ‘CBSD-like disease’. Consequently, the disease found in DRC cassava samples has been designated ‘Cassava Root Necrosis Disease’ or CRND.

Allelopathy of Dahlstedtia araripensis on Calotropis procera and Zea mays

Studies related to the allelopathic properties of plants have aroused great interest, since species that have compounds with allelopathic activity can be used as bioherbicides in the control of weeds. In this way, the aim of this study was to evaluate the allelopathic action of Dahlstedtia araripensis on the germination and growth of Calotropis procera and Zea mays. The bioassays were prepared using two 50 g portions of leaves, stem bark and D. araripensis roots, and each part of the plant received a hot treatment (1 L of distilled water at 100 °C) and one part cold (1 L of distilled water at 25 °C). The experimental design consisted of six treatments and the control group. The variables analyzed were: Index of Emergency Speed (IES), germinability, length and occurrence of necrotic radicles. The results indicated that the extracts interfered negatively on the germinability of the seeds, mainly on those of C. procera, since all extracts significantly inhibited its germination. In the seeds of C. procera and Z. mays there was delay in IES. The results indicated that the cold and hot extracts of the distinct parts of D. araripensis affected the development of the seedlings, besides promoting root necrosis. The observed effects may be due to the presence of secondary metabolites detected in the different extracts of D. araripensis. However, further research is required to prove the performance of such compounds, as well as their isolation, for future use asnatural herbicides.