Screening landraces for additional sources of field resistance to cassava mosaic disease and green mite for integration into the cassava improvement program

Twelve cassava landraces were evaluated for sources of resistance genes to diseases and pests of major economic importance in Africa. The objective was to assess their levels of field resistance to mosaic disease （ACMD）, bacterial blight （CBB）, anthracnose （CAD）, and green mite （CGM）, compared to TMS30572, an elite cultivar widely adopted in Africa. Considerable genotypic variation was observed among cultivars for resistance to ACMD and CGM but not for CBB and CAD. The lowest mean incidence of 12% and severity of 1.8 on a scale of 1-5 for ACMD was recorded for Atu, a landrace with farmer acceptable qualities. In comparison, the improved cultivar, TMS 30572, had a mean disease incidence of 72% and a severity score of 2.8. Another landrace, MS-20 had the lowest CGM damage score （2.1） while TMS 30572 emerged as one of the susceptible cultivars with a damage score of 3. Additional sources of resistance to ACMD and CGM that may possibly be better than the popular improved cultivar, TMS 30572, were identified in this study. These could serve as novel sources of additional genes to complement existing resources for elite cassava breeding in Africa.     

A Screening Method for Detecting Resistance Against Cassava Bacterial Blight Disease

Inoculation of the cassava bacterial blight (CBB) pathogen by using a modified pair of tongs on cassava shoots (1-month-old) in the glasshouse and in the field resulted in symptoms within 24-48 h on susceptible cassava lines ‘02864’ and ‘M'pelolongi’. Gum exudates on inoculated young stems and die-back appeared 7-12 and 20-25 days later, respectively. No symptoms of CBB were observed on cv. ‘Kadanga Malombe’. Symptom development and CBB ratings on 3-month-old cassava shoots in the glasshouse and inthe field were not consistent. Inoculation of CBB pathogen on 521 cassava shoots (1-month-old) using the tongs method gave 81 and 82 resistant and moderately resistant plants to CBB, respectively. The tongs method is an excellent field-screening technique for detecting resistance to CBB on 1-month-old cassava shoots.     

The Relative Resistance of Cassava Cultivars to African Cassava Mosaic Disease (ACMD) as Determined by Two Methods: Rank-Sum and the Area Under the Disease Progress Curve

Twenty-five newly bred improved cassava cultivars, twenty-three improved from the International Institute of Tropical Agriculture and two local cultivars were evaluated for their relative resistance to African cassava mosaic begomovirus disease (ACMD) at Ibadan, in an area of high disease pressure representative of the forest/savanna transition zone of Nigeria. These cultivars were exposed to natural infection by the viruliferous whiteflies ( Bemisia tabaci Gennadius) and the disease incidence (DI) and index of symptom severity (ISS) were assessed for all clones. Results for the Rank-sum (i.e., sum of ranks for DI and ISS for each cultivar) and the area under the disease progress curve (AUDPC) were used to assess the relative resistance of the cassava clones. Those that showed low Rank-sum and AUDPC values were rated 'moderately resistant (MR)', 'resistant ( R )', and 'highly resistant (HR)' to ACMD depending on their respective values and deviation from the mean distribution curve. Clones M94/0121 and 94/0239 were rated HR under the two methods. Clone M94/0583 was rated HR under the AUDPC with a deviation from the mean distribution curve of m 2.00 while it was rated R under the Rank-sum method with a deviation from the mean distribution of m 1.99. Also plants of clones ISU and TMS 30572 were rated highly susceptible (HS) under both methods. Clone TME-1 was intermediate between Moderately resistant (MR) and Moderately susceptible (MS) under the AUDPC method with a deviation from the mean distribution of 0.00 but observed to be MS under the Rank-sum method with a deviation of + 0.2. The two methods of evaluation gave similar results as revealed by Spearman rank correlation ( r equals; 0.99, P <0.01). However, the AUDPC method is less cumbersome compared to the Rank-sum method. None of the clones was observed to be immune to the disease.     

Relations Between Biomass Accumulation and Infection by Vesicular-Arbuscular Mycorrhizal Fungi in Cassava as Affected by Planting Date

Roots of 12, 3-month-old, field-propagated clones of cassava (Manihot esculenta Crantz) were more heavily mycorrhizal in the dry (Jan.–March and Oct.–Dec.) (39–83 %) than in the wet season (April–Sept.) (20–71%). Mean dry weights (biomass) of roots and shoots in the wet season (3.8–7.9 and 9.7–19.1 g/plant) were higher (P = 0.01) than in the dry season (2.1–5.9 and 5.4–12.8 g/plant, respectively). Clones with symptoms of the African cassava mosaic disease (ACMD) were less mycorrhizal, (20–69%) than mosaic symptom-free clones (51–83%). Higher colonization of roots of the clones by indigenous fungal symbionts and lower biomass accumulation in the dry season are attributed mainly to soil moisture and other effects, while reduced infections in cassava with ACMD symptoms may be due to ACMD-induced reductions in carbohydrate levels.     

Rapid Screening Method of Cassava Cultivars for Resistance to Colletotrichum gloeosporioides f.sp. manihotis

An in vitro method for assessing cassava anthracnose disease (CAD) resistance was developed as a preliminary screen to a CAD-resistant breeding programme. Potato dextrose agar (PDA) media was amended by extracts from the stem cortex of 10 cassava cultivars (30001; 30572, 30211, 88/02549, 88/00695, 88/01336, 91/00344, 91/00313, 91/00684 and 91/00475), and assayed for efficacy of inhibition of the growth of Colletotrichum gloeosporioides f. sp. manihotis isolates (05FCN, 10FCN, 12FCN, and 18FCN). Morphological and physiological data indicated that there was a significant difference (P ≤ 0.05), in mycelial growth, spore germination and sporulation among the four isolates on PDA amended with cassava stem extracts. Extracts from cassava cultivars 30211, 91/00684 and 91/00313 showed higher inhibition of germ tube development, mycelial growth and sporulation of the fungal isolates, whereas cultivars 88/02549 and 88/01336 showed the least inhibition. The 10 cultivars were further tested in both greenhouse and field conditions, under disease pressure for two planting seasons, to corroborate resistance to the fungus as observed in vitro . Greenhouse and field trials with the 10 cassava cultivars showed a significant difference ( P  ≤ 0.05) in CAD resistance. Cultivars 88/02549 and 88/01336 were highly CAD-susceptible, as shown in the in vitro assays and confirmed in the greenhouse and field tests. The other eight cultivars were either resistant (30211, 91/00684), or moderately resistant (30572, 88/00695, 91/00475, 91/00344, 30001 and 91/00313) to CAD. The study shows that an in vitro screening assay of cassava for resistance to CAD could serve as a convenient preliminary screening technique to discriminate CAD-resistant from CAD-susceptible cassava cultivars. The in vitro screening method considerably reduces time and labour in comparison with the current screening techniques of cassava, which involve field planting, inoculation and evaluation.     

Cassava genetic transformation and its application in breeding

As a major source of food, cassava (Manihot esculenta Crantz) is an important root crop in the tropics and subtropics of Africa and Latin America, and serves as raw material for the production of starches and bioethanol in tropical Asia. Cassava improvement through genetic engineering not only overcomes the high heterozygosity and serious trait separation that occurs in its traditional breeding, but also quickly achieves improved target traits. Since the first report on genetic transformation in cassava in 1996, the technology has gradually matured over almost 15 years of development and has overcome cassava genotype constraints, changing from mode cultivars to farmer-preferred ones. Significant progress has been made in terms of an increased resistance to pests and diseases, biofortification, and improved starch quality, building on the fundamental knowledge and technologies related to planting, nutrition, and the processing of this important food crop that has often been neglected. Therefore, cassava has great potential in food security and bioenergy development worldwide.     

Field Studies of Cross Protection with Cassava Mosaic Geminiviruses in Uganda

The effect of cassava mosaic virus disease (CMD) was compared on plants grown from cuttings that were initially virus‐free at planting and those infected with a mild strain of East African cassava mosaic virus‐Uganda (EACMV‐UG). All initially healthy plants developed CMD symptoms within 5 months of planting (MAP) at both trial sites in Uganda, although spread was more rapid at Kamuli than at Serere. Significantly (P < 0.001) higher symptom severity scores were recorded in initially healthy plants, which had average scores of 3.6 and 3.5 at Kamuli and Serere, respectively, compared with 2.8 for mildly diseased plants at each location. Severity scores of 4 and 5 were more frequent in initially healthy plants, accounting for 77 and 39% of the total infections recorded in comparison with 47 and 11% in mildly diseased plants at Kamuli and Serere, respectively. Mildly diseased plants were significantly taller than initially healthy plants 8 and 12 MAP at both locations. However, the converse was true 4 MAP although differences were significant at Serere but not at Kamuli. Mildly diseased plants yielded significantly more tuberous roots than initially healthy plants at Kamuli but not at Serere. Average total weights of tuberous roots per plant were 2.48 and 1.63 kg for mildly diseased and initially healthy plants at Kamuli and 4.46 and 4.61 kg at Serere, respectively. These results may help to explain the increased prevalence in recent years of mildly diseased plants of local CMD‐susceptible cultivars in eastern Uganda, from where these varieties virtually disappeared following the severe CMD epidemic in the 1990s. The results also provide the first field evidence of a cross protective effect of mild strains of a cassava mosaic geminivirus.     

Use of resistant and susceptible potato cultivars in the trap cropping of potato cyst nematodes, Globodera pallida and G. rostochiensis

The effects of planting date and growing period of potato cultivars on their efficiency as trap crops for potato cyst nematodes (PCN) were studied. Plots were planted with susceptible or resistant cultivars in April, June and August and these were grown for 5, 6 or 7 wk before removal of the plants by hand lifting. Crops planted in June provided the best overall reductions in PCN population density of up to 95%, with cv. Santé significantly more effective than the other cultivars. Population reductions from the August planting were only slightly less than from planting in June but the tuber yields obtained were much greater: Maris Piper and Maris Bard produced 16.4 and 21.4 t ha^-1 respectively, with 37% and 43% respectively, of a size useful for canning (i.e. between 20 and 40 mm diameter).     

Cassava diseases caused by Xanthomonas phaseoli pv. manihotis and Xanthomonas cassavae

Xanthomonas phaseoli pv. manihotis (Xpm) and X. cassavae (Xc) are two bacterial pathogens attacking cassava. Cassava bacterial blight (CBB) is a systemic disease caused by Xpm, which might have dramatic effects on plant growth and crop production. Cassava bacterial necrosis is a nonvascular disease caused by Xc with foliar symptoms similar to CBB, but its impacts on the plant vigour and the crop are limited. In this review, we describe the epidemiology and ecology of the two pathogens, the impacts and management of the diseases, and the main research achievements for each pathosystem. Because Xc data are sparse, our main focus is on Xpm and CBB.     

The Cassava Genome: Current Progress, Future Directions

The starchy swollen roots of cassava provide an essential food source for nearly a billion people, as well as possibilities for bioenergy, yet improvements to nutritional content and resistance to threatening diseases are currently impeded. A 454-based whole genome shotgun sequence has been assembled, which covers 69% of the predicted genome size and 96% of protein-coding gene space, with genome finishing underway. The predicted 30,666 genes and 3,485 alternate splice forms are supported by 1.4 M expressed sequence tags (ESTs). Maps based on simple sequence repeat (SSR)-, and EST-derived single nucleotide polymorphisms (SNPs) already exist. Thanks to the genome sequence, a high-density linkage map is currently being developed from a cross between two diverse cassava cultivars: one susceptible to cassava brown streak disease; the other resistant. An efficient genotyping-by-sequencing (GBS) approach is being developed to catalog SNPs both within the mapping population and among diverse African farmer-preferred varieties of cassava. These resources will accelerate marker-assisted breeding programs, allowing improvements in disease-resistance and nutrition, and will help us understand the genetic basis for disease resistance.     

Control of Globodera tabacum solanacearum by Alternating Host Resistance and Nematicide

The feasibility of alternating use of resistant vs. susceptible flue-cured tobacco cultivars to improve control of Globodera tabacum subsp, solanacearum (TCN) was investigated at two Virginia locations in 1984-86. Post-harvest TCN population densities were reduced in each year of the study when fenamiphos was used with a TCN-resistant cultivar (NC 567), relative to susceptible cultivars (K 326 or Mc 944). Using NC 567 with fenamipbos also reduced preplant TCN population densities in the next growing season. Egg population densities before planting in 1986 were significantly lower in plots planted with NC 567 in 1984, even when a susceptible cultivar had been planted in 1985. Use of fenamiphos with NC 567 in 1984 and 1985 further reduced preplant egg population densities in 1986. Economic returns were significantly greater in 1984 when NC 567 was used with fenamiphos, rather than a susceptible cultivar. Treatments involving fenamiphos and (or) NC 567 in 1984 and 1985 resulted in higher economic returns in 1986 than did treatments using a susceptible cultivar without fenamiphos in both previous years. Economic returns were highest in 1986 when fenamiphos and NC 567 were used in 1984 and 1985 and a susceptible cultivar was planted in 1986.     

Endophytic bacterial and fungal microbiota in different cultivars of cassava (Manihot esculenta Crantz)

Endophytes colonize tissues of healthy host plants and play a crucial role in plant growth and development. However, little attention has been paid to the endophytes of tuber crops such as cassava, which is used as a staple food by approximately 800 million people worldwide. This study aimed to elucidate the diversity and composition of endophytic bacterial and fungal communities in different cassava cultivars using high-throughput sequencing. Although no significant differences in richness or diversity were observed among the different cassava cultivars, the community compositions were diverse. Two cultivars (SC124 and SC205) tolerant to root rot exhibited similar community compositions, while two other cultivars (SC10 and SC5), which are moderately and highly susceptible to root rot, respectively, harboured similar community compositions. Proteobacteria, Firmicutes, and Ascomycota dominated the endophyte assemblages, with Weissella, Serratia, Lasiodiplodia, Fusarium, and Diaporthe being the predominant genera. The differentially abundant taxonomic clades between the tolerant and susceptible cultivars were mainly rare taxa, such as Lachnoclostridium_5, Rhizobium, Lampropedia, and Stenotrophomonas. These seemed to be key genera that affected the susceptibility of cassava to root rot. Moreover, the comparison of KEGG functional profiles revealed that ‘Environmental adaptation’ category was significantly enriched in the tolerant cultivars, while ‘Infectious diseases: Parasitic’ category was significantly enriched in the susceptible cultivars. The present findings open opportunities for further studies on the roles of endophytes in the susceptibility of plants to diseases.

     

The influence of growth stages on cassava tuberous root rot in different ecological environments

A multilocational field experiment to determine the influence of cassava growth and development on tuberous root rots in Onne and Sabongidda-Ora humid forests and in the Ibadan derived savanna of Nigeria was carried out from June 2003 to November 2005. Healthy cassava stems of TME-1 (local), TMS 30572 and TMS 4 (2) 1425 (both improved) were planted in a randomised complete block design with three replications. The experiment was repeated and the data collected were statistically analysed using the general linear model statistical procedures with the SAS system for Windows. Experimental result showed tuberous root rot incidence and severity to be pronounced at 12 months after planting in Onne and Sabongidda-Ora humid forests but less at the Ibadan derived savanna. Local TME-1 variety had the highest root rot incidence of 53% and severity of 21% while improved TMS 4(2)1425 variety had the least root rot incidence and severity at 0% each.     

The Spread of African Cassava Mosaic Virus into and within Cassava Fields

Only a small proportion of Bemisia tabaci collected in totally infected cassava fields at a site in Côte d'lvoire transmitted African cassava mosaic virus (ACMV) to test plants. Nevertheless, the monthly increase in disease incidence in an experimental planting was directly related to numbers of adult whiteflies counted on plants 6 weeks earlier. In plots at different spacings, the greatest incidence of ACMV expressed as a percentage of the total stand occurred at the lowest plant density. Much speread into the spacing trial and into two other experimental plantings occurred from outside sources and followed downwind gradients. By contrast, spread from ACMV-infected sources within plantings was limited. It occurred in all directions but over distances of only a few metres. These contrasting patterns of spread are attributed to the differentbehavior of B. tabaci above and within the crop canopy. It is concluded that contamination of cassava fields in the coastal forest area of Côte d'Ivoire is due mainly to rapid spread from outside sources which leads to internal foci that contribute to some further, although limited, spread. These findings are discussed in relation to possible control strategies based on the release of healthy cuttings, dense planting and subsequent roguing. Such measures are unlikely to be effectve in the coastal forest region of Côte d'Ivoire and adjacent countriesunless varieties are grown with greater resistance to infection than those currently used.     

Tissue Culture of Cassava on Chemically Defined Media

Defined media that promote the initiation and undifferentiated growth of callus derived from stem explants of four cultivars of cassava, Manihot esculenta Crantz, are described. Growth rates and yields of cassava callus after 4 weeks of culture are shown to be comparable to those of callus of Nicotiana tabacum L. cv. Wisconsin No. 38. Nitrogen sources of ammonium nitrate or of ammonium chloride plus succinate supported growth of all four cultivars. Sucrose was superior to glucose as a carbon source. The cassava cultivars differed in their response to increasing concentrations of sucrose between 0.5% (w/v) and 3%, two of them increasing in dry matter with increasing sucrose concentrations of up to 3%. When cultured in the light on defined media that contained higher ratios of cytokinin to auxin, callus of the latter two cultivars turned green. Roots but not shoots differentiated from the callus of all cultivars. The influence of hormone concentrations, sucrose level, and nitrogen source on greening and root formation is summarized.     

Evaluation of Cassava Cultivars for Resistance to Cassava Mosaic Disease and Yield Potential in Central African Republic

Eleven cassava genotypes were tested against cassava mosaic disease (CMD) and compared to a local susceptible cultivar in field conditions from June 2011 to July 2012 in Central African Republic (CAR) at two sites representative of the savanna (Damara) and forest (Pissa) zones of the country. The mean number of whiteflies observed on plants varied among genotypes within each site, but was found nearly three times higher at Damara than at Pissa, resulting in a CMD incidence nearly five times higher at Damara than at Pissa. However, no relation was observed between the number of insect on the plants and the level of susceptibility/resistance of the genotypes. The difference of disease pressure between the two sites revealed high level of resistance in several genotypes, while some other ones indicated rather only a partial resistance. Nevertheless, none of the genotypes tested was found immune, in the end, the virus being detected at least in one site in every genotype, including those ones presenting no symptoms in both sites. The impact of CMD on yield components was assessed on the local susceptible check and three partially resistant genotypes, showing that the disease has no significant effect on the tuberous roots number as well as their weight in both sites. The yield potential varied among different genotypes and between the two sites, the mean number of tuberous roots as well as their mean weight being higher in Damara than in Pissa. This study identified highly resistant genotypes such as ‘Gabon’ that performed well in both sites, and ‘91/02322’ that was symptomless and presented a yield potential equivalent to the local check. These genotypes could be distributed to growers with the main advantage to be resistant to CMD and, therefore, reducing the risk to spread sources of inoculum all over the cassava cropping areas in the country.     

Screening of cassava varieties for resistance to root-knot nematodes

Twenty cassava cultivars were screened in micro-plot experiments in the green house of Institute of Agricultural Research and Training, Obafemi Awolowo University, Moor Plantation, Ibadan for root-knot nematode resistance. Four weeks after planting, each plant was inoculated with 5000 eggs of Meloidogyne incognita. There were three replications per cultivar. The plants were uprooted and assessed for resistance after 60 days of inoculation. Assessment of plant damaged was determined by gall index and reproductive factor. Based on host status rating, 3 cultivars were found to be resistant, 4 cultivars were tolerant, 10 cultivars ere susceptible while .3 cultivars were hypersusceptible. All the screened cultivars are resistant to cassava mosaic disease and other major pests of cassava like bacterial disease, anthracnose disease, cassava green mite and cassava mealy bug. They are all high yielding, suitable for food, industry and livestock feed.     

Viruses Associated with Cassava Mosaic Disease in Senegal and Guinea Conakry

A survey in Senegal and Guinea Conakry established the presence and incidence of cassava mosaic virus disease (CMD) in both countries. CMD occurred in all the fields surveyed, although its incidence was higher in Senegal (83%) than in Guinea (64%). Populations of the whitefly vector, Bemisia tabaci, were low in both countries averaging 1.7 adults per shoot in Guinea and 3.2 in Senegal. Most infections were attributed to the use of infected cuttings, 86 and 83% in Senegal and Guinea, respectively, and there was no evidence of rapid current‐season, whitefly‐borne infection at any of the sampled locations. Disease severity was generally low in the two countries and averaged 2.5 in Guinea and 2.3 in Senegal. No plants with unusually severe CMD symptoms characteristic of the CMD pandemic in East and Central Africa were observed. Restriction fragment length polymorphism (RFLP)‐based diagnostics revealed that African cassava mosaic virus (ACMV) is exclusively associated with CMD in both the countries. Neither East African cassava mosaic virus (EACMV), nor the recombinant Uganda variant (EACMV‐UG2) was detected in any sample. These survey data indicate that CMD could be effectively controlled in both countries by phytosanitation, involving the use of CMD‐free planting material and the removal of diseased plants.