Assessing the Degeneration of Cassava Under High-Virus Inoculum Conditions in Coastal Tanzania.

Cassava brown streak disease (CBSD), caused by cassava brown streak ipomoviruses (CBSIs), has become the most debilitating biotic stress to cassava production in East and Central Africa. Lack of CBSD-resistant varieties has necessitated the search for alternative control measures. Most smallholder farmers reuse stems from previous crops for planting in the new season. Recycling planting material in this way can lead to "degeneration" owing to the compounding effects of disease. In this study, degeneration was defined as the increase in CBSD incidence and reduction in marketable root yield over time. An experiment was established to study the rates of degeneration in selected cassava varieties Chereko, KBH2002_135, Kipusa, Kizimbani, and Mkuranga1 and cultivars Kiroba and Kikombe under high-CBSD inoculum conditions in Bagamoyo, Tanzania from 2013 to 2017. The experiment was replicated across two seasons: the first planted during the long rains (Masika) between March and June and the second planted during the short rains (Vuli) between October and December. Mean abundance of the whitefly vector (Bemisia tabaci) was much greater during the Vuli season (>19 insects per plant) than the Masika season (<2 insects per plant). CBSD shoot symptoms occurred naturally and were observed only on Kikombe, Kiroba, and Kipusa. New materials had overall lower CBSD shoot incidences (1.5%) compared with recycled materials (6.9%) in Masika, although no significant differences were obvious in Vuli. However, Masika (8.7%) had an overall lower CBSD shoot incidence than Vuli (16.5%) in the varieties that had shoot symptoms. CBSD root incidences were higher in Vuli (10.3%) than Masika (4.4%), and root yields in Masika (29.4 t/ha) were significantly greater than those in Vuli (22.5 t/ha). The highest percentage of roots rendered unusable owing to CBSD was observed in Vuli. There was significantly higher unusable root incidence in recycled materials (3.7%) than in new materials (1.4%) in Masika but not in Vuli. Overall root yield was similar between recycled and new materials in either season. Significant reductions in root yield over the course of the experiment were observed both in Masika and Vuli, whereas changes in marketable yield were significant only in Masika. Differences in the response of varieties to degeneration led to the identification of four degeneration patterns, namely "strong," "moderate," "mild," and "delayed" degeneration. The strongest effects of degeneration were most obvious in the susceptible cultivar (Kikombe), which also had the lowest marketable yield in either season. Seasonal differences were a key driver of degeneration, because its effects were much greater in Vuli than Masika. To the best of our knowledge, this work reports the first study of degeneration caused by cassava viruses.[Formula: see text] Copyright © 2019 The Author(s). This is an open access article distributed under the CC BY 4.0 International license.

Genotype by environment cultivar evaluation for cassava brown streak disease resistance in Tanzania.

Cassava brown steak disease (CBSD), caused by Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV), is the most important biotic constraint to cassava production in East and Central Africa. Concerted efforts are required to prevent further spread into West Africa as well as to reduce losses in areas already affected. The study reported here was part of a five-country (Kenya, Malawi, Mozambique, Tanzania and Uganda) programme that aimed to identify superior cassava cultivars resistant to CBSD and to disseminate them widely in the region. Seventeen tissue-cultured and virus-tested cultivars were evaluated in Tanzania across nine sites with diverse CBSD inoculum conditions. Experiments were planted using an alpha-lattice design and assessments were made of surrounding inoculum pressure, CBSD foliar and root incidence and root yield at harvest. There were large differences in CBSD infection between sites, with greatest spread recorded from the north-western Lake (Victoria) zone. Differences were driven by Bemisia tabaci whitefly vector abundance and CBSD inoculum pressure. Both CBSV and UCBSV were almost equally represented in cassava fields surrounding experimental plots, although CBSV predominated in the north-west whilst UCBSV was more frequent in coastal and southern sites. However, the incidence of CBSV was much greater than that of UCBSV in initially virus-free experimental plots, suggesting that CBSV is more virulent. Cultivars could be categorised into three groups based on the degree of CBSD symptom expression in shoots and roots. The seven cultivars (F10_30R2, Eyope, Mkumba, Mkuranga1, Narocass1, Nase3 and Orera) in the most resistant category each had shoot and root incidences of less than 20%. Fresh root yield differed between sites and cultivars, but there was no genotype by environment interaction for this trait, probably attributable to the large fertility and soil moisture differences between sites. Susceptible cultivars and the local check performed well in the absence of CBSD pressure, highlighting the importance of exploiting quality and yield traits of local landraces in breeding programmes. Overall, our results emphasized the importance of applying a balanced strategy for CBSD management. This should use both improved and local germplasm resources to generate high yielding cultivars for specific end-user traits, and combine the deployment of improved cultivars with phytosanitary control measures including the use of healthy planting material and planting during periods of reduced CBSD infection.

Absolute quantification of cassava brown streak virus mRNA by real-time qPCR.

Cassava brown streak disease (CBSD) is the most important virus disease of cassava and a major food security threat in Africa. Yearly economic losses of up to $100 million USD have been attributed to CBSD. The lack of information on plant-virus interactions has restricted progress in breeding for CBSD resistance. Virus quantification is becoming a major tool for the quick and reliable assessment of plant host resistance. Therefore, a protocol for specific absolute quantification of Cassava brown streak virus (CBSV) and Ugandan cassava brown streak virus (UCBSV) was developed. CBSV and UCBSV coat protein (CP) specific standard templates: CBSV (pFer2, 826bp) and UCBSV (pUF1-R1-1, 732) respectively were generated and maintained in a TA cloning vector. These were used to construct standard curves using a TaqMan qPCR assay. Standard curves with acceptable amplification efficiencies (90-105%) and coefficients of determination (R2) greater than 0.99 were obtained. Infected cassava plants were sampled from a screenhouse and the field and used to validate this assay. Results obtained by testing several screenhouse and field samples revealed consistent absolute quantification assays for different CBSV and UCBSV isolates. This study presents the first protocol for absolute quantification of CBSVs and is expected to accelerate screening for CBSD resistance and hence breeding for CBSD resistance. The use of the method presented here should improve the clarity of virus quantification data as the results obtained are not influenced by varietal, host, seasonal or environmental conditions. Screening efficiency will also be greatly improved as there is no need for the use of reference genes consequently allowing for a larger number of samples to be analyzed. This will increase experimental precision in a timely and cost effective manner.