The Significance of Wild Plants in the Evolutionary Ecology of Three Major Viruses Infecting Cultivated Sweetpotato in Uganda

The studies presented in this thesis contribute to the understanding of evolutionary ecology of three major viruses threatening cultivated sweetpotato (Ipomoea batatas Lam) in East Africa: Sweet potato feathery mottle virus (SPFMV; genus Potyvirus; Potyviridae), Sweet potato chlorotic stunt virus (SPCSV; genus Crinivirus; Closteroviridae) and Sweet potato mild mottle virus (SPMMV; genus Ipomovirus; Potyviridae). The viruses were serologically detected and the positive results confirmed by RT-PCR and sequencing. SPFMV was detected in 24 wild plant species of family Convolvulacea (genera Ipomoea, Lepistemon and Hewittia), of which 19 species were new natural hosts for SPFMV. SPMMV and SPCSV were detected in wild plants belonging to 21 and 12 species (genera Ipomoea, Lepistemon and Hewittia), respectively, all of which were previously unknown to be natural hosts of these viruses. SPFMV was the most abundant virus being detected in 17% of the plants, while SPMMV and SPCSV were detected in 9.8% and 5.4% of the assessed plants, respectively. Wild plants in Uganda were infected with the East African (EA), common (C), and the ordinary (O) strains, or co-infected with the EA and the C strain of SPFMV. The viruses and virus-like diseases were more frequent in the eastern agro-ecological zone than the western and central zones, which contrasted with known incidences of these viruses in sweetpotato crops, except for northern zone where incidences were lowest in wild plants as in sweetpotato. The NIb/CP junction in SPMMV was determined experimentally which facilitated CP-based phylogenetic and evolutionary analyses of SPMMV. Isolates of all the three viruses from wild plants were genetically similar to those found in cultivated sweetpotatoes in East Africa. There was no evidence of host-driven population genetic structures suggesting frequent transmission of these viruses between their wild and cultivated hosts. The p22 RNA silencing suppressor-encoding sequence was absent in a few SPCSV isolates, but regardless of this, SPCSV isolates incited sweet potato virus disease (SPVD) in sweetpotato plants co-infected with SPFMV, indicating that p22 is redundant for synergism between SCSV and SPFMV. Molecular evolutionary analysis revealed that isolates of strain EA of SPFMV that is largely restricted geographically in East Africa experience frequent recombination in comparison to isolates of strain C that is globally distributed. Moreover, non-homologous recombination events between strains EA and C were rare, despite frequent co-infections of these strains in wild plants, suggesting purifying selection against non-homologous recombinants between these strains or that such recombinants are mostly not infectious. Recombination was detected also in the 5 - and 3 -proximal regions of the SPMMV genome providing the first evidence of recombination in genus Ipomovirus, but no recombination events were detected in the characterized genomic regions of SPCSV. Strong purifying selection was implicated on evolution of majority of amino acids of the proteins encoded by the analyzed genomic regions of SPFMV, SPMMV and SPCSV. However, positive selection was predicted on 17 amino acids distributed over the whole the coat protein (CP) in the globally distributed strain C, as compared to only 4 amino acids in the multifunctional CP N-terminus (CP-NT) of strain EA largely restricted geographically to East Africa. A few amino acid sites in the N-terminus of SPMMV P1, the p7 protein and RNA silencing suppressor proteins p22 and RNase3 of SPCSV were also submitted to positive selection. Positively selected amino acids may constitute ligand-binding domains that determine interactions with plant host and/or insect vector factors. The P1 proteinase of SPMMV (genus Ipomovirus) seems to respond to needs of adaptation, which was not observed with the helper component proteinase (HC-Pro) of SPMMV, although the HC-Pro is responsible for many important molecular interactions in genus Potyvirus. Because the centre of origin of cultivated sweetpotato is in the Americas from where the crop was dispersed to other continents in recent history (except for the Australasia and South Pacific region), it would be expected that identical viruses and their strains occur worldwide, presuming virus dispersal with the host. Apparently, this seems not to be the case with SPMMV, the strain EA of SPFMV and the strain EA of SPCSV that are largely geographically confined in East Africa where they are predominant and occur both in natural and agro-ecosystems. The geographical distribution of plant viruses is constrained more by virus-vector relations than by virus-host interactions, which in accordance of the wide range of natural host species and the geographical confinement to East Africa suggest that these viruses existed in East African wild plants before the introduction of sweetpotato. Subsequently, these studies provide compelling evidence that East Africa constitutes a cradle of SPFMV strain EA, SPCSV strain EA, and SPMMV. Therefore, sweet potato virus disease (SPVD) in East Africa may be one of the examples of damaging virus diseases resulting from exchange of viruses between introduced crops and indigenous wild plant species. Keywords: Convolvulaceae, East Africa, epidemiology, evolution, genetic variability, Ipomoea, recombination, SPCSV, SPFMV, SPMMV, selection pressure, sweetpotato, wild plant species Author s Address: Arthur K. Tugume, Department of Agricultural Sciences, Faculty of Agriculture and Forestry, University of Helsinki, Latokartanonkaari 7, P.O Box 27, FIN-00014, Helsinki, Finland. Email: tugume.arthur@helsinki.fi Author s Present Address: Arthur K. Tugume, Department of Botany, Faculty of Science, Makerere University, P.O. Box 7062, Kampala, Uganda. Email: aktugume@botany.mak.ac.ug, tugumeka@yahoo.com

Legume pastures can reduce N2O emissions intensity in subtropical cereal cropping systems

Alternative sources of N are required to bolster subtropical cereal production without increasing N2O emissions from these agro-ecosystems. The reintroduction of legumes in cereal cropping systems is a possible strategy to reduce synthetic N inputs but elevated N2O losses have sometimes been observed after the incorporation of legume residues. However, the magnitude of these losses is highly dependent on local conditions and very little data are available for subtropical regions. The aim of this study was to assess whether, under subtropical conditions, the N mineralised from legume residues can substantially decrease the synthetic N input required by the subsequent cereal crop and reduce overall N2O emissions during the cereal cropping phase. Using a fully automated measuring system, N2O emissions were monitored in a cereal crop (sorghum) following a legume pasture and compared to the same crop in rotation with a grass pasture. Each crop rotation included a nil and a fertilised treatment to assess the N availability of the residues. The incorporation of legumes provided enough readily available N to effectively support crop development but the low labile C left by these residues is likely to have limited denitrification and therefore N2O emissions. As a result, N2O emissions intensities (kgN2O-N yield-1ha-1) were considerably lower in the legume histories than in the grass. Overall, these findings indicate that the C supplied by the crop residue can be more important than the soil NO3 - content in stimulating denitrification and that introducing a legume pasture in a subtropical cereal cropping system is a sustainable practice from both environmental and agronomic perspectives.

Denitrification losses from an intensively managed sub-tropical pasture: Impact of soil moisture on the partitioning of N2 and N2O emissions

Intensively managed pastures in subtropical Australia under dairy production are nitrogen (N) loaded agro-ecosystems, with an increased pool of N available for denitrification. The magnitude of denitrification losses and N2:N2O partitioning in these agro-ecosystems is largely unknown, representing a major uncertainty when estimating total N loss and replacement. This study investigated the influence of different soil moisture contents on N2 and N2O emissions from a subtropical dairy pasture in Queensland, Australia. Intact soil cores were incubated over 15 days at 80% and 100% water-filled pore space (WFPS), after the application of 15N labelled nitrate, equivalent to 50 kg N ha−1. This setup enabled the direct quantification of N2 and N2O emissions following fertilisation using the 15N gas flux method. The main product of denitrification in both treatments was N2. N2 emissions exceeded N2O emissions by a factor of 8 ± 1 at 80% WFPS and a factor of 17 ± 2 at 100% WFPS. The total amount of N-N2 lost over the incubation period was 21.27 kg ± 2.10 N2-N ha−1 at 80% WFPS and 25.26 kg ± 2.79 kg ha−1 at 100% WFPS respectively. N2 emissions remained high at 100% WFPS, while related N2O emissions decreased. At 80% WFPS, N2 emissions increased constantly over time while N2O fluxes declined. Consequently, N2/(N2 + N2O) product ratios increased over the incubation period in both treatments. N2/(N2 + N2O) product ratios responded significantly to soil moisture, confirming WFPS as a key driver of denitrification. The substantial amount of fertiliser lost as N2 reveals the agronomic significance of denitrification as a major pathway of N loss for sub-tropical pastures at high WFPS and may explain the low fertiliser N use efficiency observed for these agro-ecosystems.

Legume pastures can reduce N2O emissions intensity in subtropical cereal cropping systems

Alternative sources of N are required to bolster subtropical cereal production without increasing N2O emissions from these agro-ecosystems. The reintroduction of legumes in cereal cropping systems is a possible strategy to reduce synthetic N inputs but elevated N2O losses have sometimes been observed after the incorporation of legume residues. However, the magnitude of these losses is highly dependent on local conditions and very little data are available for subtropical regions. The aim of this study was to assess whether, under subtropical conditions, the N mineralised from legume residues can substantially decrease the synthetic N input required by the subsequent cereal crop and reduce overall N2O emissions during the cereal cropping phase. Using a fully automated measuring system, N2O emissions were monitored in a cereal crop (sorghum) following a legume pasture and compared to the same crop in rotation with a grass pasture. Each crop rotation included a nil and a fertilised treatment to assess the N availability of the residues. The incorporation of legumes provided enough readily available N to effectively support crop development but the low labile C left by these residues is likely to have limited denitrification and therefore N2O emissions. As a result, N2O emissions intensities (kg N2O-N yield−1 ha−1) were considerably lower in the legume histories than in the grass. Overall, these findings indicate that the C supplied by the crop residue can be more important than the soil NO3− content in stimulating denitrification and that introducing a legume pasture in a subtropical cereal cropping system is a sustainable practice from both environmental and agronomic perspectives.

Impact of compost, vermicompost and biochar on soil fertility, maize yield and soil erosion in Northern Vietnam: A three year mesocosm experiment

Compost, vermicompost and biochar amendments are thought to improve soil quality and plant yield. However, little is known about their long-term impact on crop yield and the environment in tropical agro-ecosystems. In this study we investigated the effect of organic amendments (buffalo manure, compost and verrnicompost) and biochar (applied alone or with vermicompost) on plant yield, soil fertility, soil erosion and water dynamics in a degraded Acrisol in Vietnam. Maize growth and yield, as well as weed growth, were examined for three years in terrestrial mesocosms under natural rainfall. Maize yield and growth showed high inter-annual variability depending on the organic amendment. Vermicompost improved maize growth and yield but its effect was rather small and was only significant when water availability was limited (year 2). This suggests that vermicompost could be a promising substrate for improving the resistance of agrosystems to water stress. When the vermicompost biochar mixture was applied, further growth and yield improvements were recorded in some cases. When applied alone, biochar had a positive influence on maize yield and growth, thus confirming its interest for improving long-term soil productivity. All organic amendments reduced water runoff, soil detachment and NH4+ and NO3- transfer to water. These effects were more significant with vermicompost than with buffalo manure and compost, highlighting that the beneficial influence of vermicompost is not limited to its influence on plant yield. In addition, this study showed for the first time that the combination of vermicompost and biochar may not only improve plant productivity but also reduce the negative impact of agriculture on water quality. (C) 2015 Elsevier B.V. All rights reserved.

Fingerprinting environmental conditions and related stress using stable isotopic composition of rice (Oryza sativa L.) grain organic matter

Sea level rise (SLR) is a primary factor responsible for inundation of low-lying coastal regions across the world, which in turn governs the agricultural productivity. In this study, rice (Oryza sativa L.) cultivated seasonally in the Kuttanad Wetland, a SLR prone region on the southwest coast of India, were analysed for oxygen, hydrogen and carbon isotopic ratios (delta O-18, delta H-2 and delta C-13) to distinguish the seasonal environmental conditions prevalent during rice cultivation. The region receives high rainfall during the wet season which promotes large supply of fresh water to the local water bodies via the rivers. In contrast, during the dry season reduced river discharge favours sea water incursion which adversely affects the rice cultivation. The water for rice cultivation is derived from regional water bodies that are characterised by seasonal salinity variation which co-varies with the delta O-18 and delta H-2 values. Rice cultivated during the wet and the dry season bears the isotopic imprints of this water. We explored the utility of a mechanistic model to quantify the contribution of two prominent factors, namely relative humidity and source water composition in governing the seasonal variation in oxygen isotopic composition of rice grain OM. delta C-13 values of rice grain OM were used to deduce the stress level by estimating the intrinsic water use efficiency (WUEi) of the crop during the two seasons. 1.3 times higher WUE, was exhibited by the same genotype during the dry season. The approach can be extended to other low lying coastal agro-ecosystems to infer the growth conditions of cultivated crops and can further be utilised for retrieving paleo-environmental information from well preserved archaeological plant remains. (c) 2015 Elsevier Ltd. All rights reserved.

Diversification of aquaculture for empowerment to fisheries through institution village linkage programme (IVLP) in Kerala, India

Technology Assessment and Refinement through the Institution Village Linkage Programme (IVLP) is the latest participatory extension model successfully undertaken by the Indian Council of Agricultural Research in India. The Central Marine Fisheries Research Institute has been implementing IVLP since 2001 to assess and refine the technologies of the coastal agro ecosystems at Elamkunnapuzha village (Vypeen Island) in the Ernakulam District of Kerala. A series of need based location specific technology intervention plans have been introduced to overcome the social and biological constraints on farming practices in fisheries, livestock and agriculture, and implemented with the active participation of the stakeholders. The inferences drawn from IVLP ultimately form a package suitable for enhanced production in the costal agro ecosystem for replication to other areas with similar characteristics. This paper gives a brief account of the treatment packages applied in fisheries through various technological interventions and discusses the consequent yield and benefits obtained. The ‘integrated whole village development’ through the involvement of multi institutional teams and a participatory approach was accorded prime importance in the IVLP of Elamkunnapuzha, with a greater emphasis on marginal and small farmers and specifically focusing on women for poverty alleviation and equity under the coastal agro ec

Assessing the consequence of land use change on agricultural productivity in China

China's cultivated land has been undergoing dramatic changes along with its rapidly growing economy and population. The impacts of land use transformation on food production at the national scale, however, have been poorly understood due to the lack of detailed spatially explicit agricultural productivity information on cropland change and crop productivity. This study evaluates the effect of the cropland transformation on agricultural productivity by combining the land use data of China for the period of 1990-2000 from TM images and a satellite-based NPP (net primary production) model driven with NOAH/AVHRR data. The cropland area of China has a net increase of 2.79 Mha in the study period, which causes a slightly increased agricultural productivity (6.96 Mt C) at the national level. Although the newly cultivated lands compensated for the loss from urban expansion, but the contribution to production is insignificant because of the low productivity. The decrease in crop production resulting from urban expansion is about twice of that from abandonment of arable lands to forests and grasslands. The productivity of arable lands occupied by urban expansion was 80% higher than that of the newly cultivated lands in the regions with unfavorable natural conditions. Significance of cropland transformation impacts is spatially diverse with the differences in land use change intensity and land productivity across China. The increase in arable land area and yet decline in land quality may reduce the production potential and sustainability of China's agro-ecosystems. (C) 2008 Elsevier B.V. All rights reserved.

Mating behavior and chemical communication in the order hymenoptera

Insects of the order Hymenoptera are biologically and economically important members of natural and agro ecosystems and exhibit diverse biologies, mating systems, and sex pheromones. We review what is known of their sex pheromone chemistry and function, paying particular emphasis to the Hymenoptera Aculeata (primarily ants, bees, and sphecid and vespid wasps), and provide a framework for the functional classification of their sex pheromones. Sex pheromones often comprise multicomponent blends derived from numerous exocrine tissues, including the cuticle. However, very few sex pheromones have been definitively characterized using bioassays, in part because of the behavioral sophistication of many Aculeata. The relative importance of species isolation versus sexual selection in shaping sex pheromone evolution is still unclear. Many species appear to discriminate among mates at the level of individual or kin/colony, and they use antiaphrodisiacs. Some orchids use hymenopteran sex pheromones to dupe males into performing pseudocopulation, with extreme species specificity.

Persistent negative effects of pesticides on biodiversity and biological control potential on European farmland

During the last 50 years, agricultural intensification has caused many wild plant and animal species to go extinct regionally or nationally and has profoundly changed the functioning of agro-ecosystems. Agricultural intensification has many components, such as loss of landscape elements, enlarged farm and field sizes and larger inputs of fertilizer and pesticides. However, very little is known about the relative contribution of these variables to the large-scale negative effects on biodiversity. In this study, we disentangled the impacts of various components of agricultural intensification on species diversity of wild plants, carabids and ground-nesting farmland birds and on the biological control of aphids.

Causes of legume-rotation effects in increasing cereal yields across the Sudanian, Sahelian and Guinean zone of West Africa

On-farm experiments and pot trials were conducted on eight West African soils to explore the mechanisms governing the often reported legume rotation-induced cereal growth increases in this region. Crops comprised pearl millet (Pennisetum glaucum L.), sorghum (Sorghum bicolor Moench), maize (Zea mays L.), cowpea (Vigna unguiculata Walp.) and groundnut (Arachis hypogaea L.). In groundnut trials the observed 26 to 85% increases in total dry matter (TDM) of rotation cereals (RC) compared with continuous cereals (CC) in the 4th year appeared to be triggered by site- and crop-specific early season differences in nematode infestation (up to 6-fold lower in RC than in CC), enhanced Nmin and a 7% increase in mycorrhizal (AM) infection. In cowpea trials yield effects on millet and differences in nematode numbers, Nmin and AM were much smaller. Rhizosphere studies indicated effects on pH and acid phosphatase activity as secondary causes for the observed growth differences between RC and CC. In the study region legume-rotation effects on cereals seemed to depend on the capability of the legume to suppress nematodes and to enhance early N and P availability for the subsequent cereal.

Effects of grassland conversion and tillage intensities on soil microbial biomass, residues and community structure

Agricultural intensification has a strong impact on level of soil organic matter (SOM), microbial biomass stocks and microbial community structure in agro-ecosystems. The size of the microbial necromass C pool could be about 40 times that of the living microbial biomass C pool in soils. Due to the specificity, amino sugar analysis gives more important information on the relative contribution of fungal and bacterial residues to C sequestration potential of soils. Meanwhile, the relationship between microbial biomass and microbial necromass in soil and its ecological significance on SOM are not fully understood and likely to be very complex in grassland soils. This thesis focuses on the effects of tillage, grassland conversion intensities and fertilisation on microbial biomass, residues and community structure. The combined analyses of microbial biomass and residue formation of both fungi and bacteria provided a unique opportunity to study the effect of tillage, grassland conversion and fertilisation on soil microbial dynamics. In top soil at 0-30 cm layer, a reduction in tillage intensity by the GRT and NT treatments increased the accumulation of saprotrophic fungi in comparison with the MBT treatment. In contrast, the GRT and NT treatments promoted AMF at the expense of saprotrophic fungi in the bottom soil layer at 30-40 cm depth. The negative relationship between the ergosterol to microbial biomass C ratio and the fungal C to bacterial C ratio points to the importance of the relationship between saprotrophic fungi and biotrophic AMF for tillage-induced changes in microbial turnover of SOC. One-season cultivation of winter wheat with two tillage events led to a significant loss in SOC and microbial biomass C stocks at 0-40 cm depth in comparison with the permanent grassland, even 5 years after the tillage event. However, the tillage induced loss in microbial biomass C was roughly 40% less in the long-term than in the short-term of the current experiment, indicating a recovery process during grassland restoration. In general, mould board tillage and grassland conversion to maize monoculture promoted saprotrophic fungi at the expense of biotrophic AMF and bacteria compared to undisturbed grassland soils. Slurry application promoted bacterial residues as indicated by the decreases in both, the ergosterol to microbial biomass C ratio and the fungal C to bacterial C ratio. In addition, the lost microbial functional diversity due to tillage and maize monoculture was restored by slurry application both in arable and grassland soils. I conclude that the microbial biomass C/S ratio can be used as an additional indicator for a shift in microbial community. The strong relationships between microbial biomass and necromass indices points to the importance of saprotrophic fungi and biotrophic AMF for agricultural management induced effects on microbial turnover and ecosystem C storage. Quantitative information on exact biomass estimates of these two important fungal groups in soil is inevitably necessary to understand their different roles in SOM dynamics.

Vigilancia tecnológica de los tres principales productos agrícolas base de alimentación y cultivados en Brasil y Argentina

La seguridad agroalimentaria debe ser prioridad para los gobiernos de Brasil y Argentina, debido a que deben garantizar la producción y el abastecimiento de los alimentos básicos para las futuras generaciones, entendido como alimentos básicos aquellos que más se consumen, ya sea en su forma original o por medio de sus subproductos, estos productos son el trigo, el maíz y el arroz. El garantizar la producción y el abastecimiento de estos productos en el corto y largo plazo, implica entender cuáles de los procesos productivos aplicados en la realización de los productos base de alimentación no son compatibles con el medio ambiente, generando impactos negativos sobre este. Estos impactos ambientales generados a partir de la agricultura, son identificados como el uso excesivo de recursos naturales entre ellos el agua, así como su contaminación por agentes toxico como los agroquímicos y fertilizantes. Asimismo, el uso de estos agentes tóxicos, genera la infertilidad de los suelos afectando directamente la producción en el largo plazo. Entender los impactos ambientales, implica desarrollar estrategias transversales que le permitan a los garantizar un desarrollo sostenible a lo largo de todo el ciclo del producto; estas estrategias deben estar acompañadas por un rendimiento y eficiencia de los cultivos, de nada sirve implementar estrategias compatibles con el medio ambiente si no se cumple con el principal objetivo de la producción de estos producto, que es garantizar el abastecimiento y alimentación para las generaciones presentes y futuras.

Arthropod diversity and community structure in relation to land use in the mekong delta, vietnam

Declining biodiversity in agro-ecosystems, caused by intensification of production or expansion of monocultures, is associated with the emergence of agricultural pests. Understanding how land-use and management control crop-associated biodiversity is, therefore, one of the key steps towards the prediction and maintenance of natural pest-control. Here we report on relationships between land-use variables and arthropod community attributes (for example, species diversity, abundance and guild structure) across a diversification gradient in a rice-dominated landscape in the Mekong delta, Vietnam. We show that rice habitats contained the most diverse arthropod communities, compared with other uncultivated and cultivated land-use types. In addition, arthropod species density and Simpson's diversity in flower, vegetable and fruit habitats was positively related to rice cover in the local landscape. However, across the landscape as a whole, reduction in heterogeneity and the amount of uncultivated cover was associated, generally, with a loss of diversity. Furthermore, arthropod species density in tillering and flowering stages of rice was positively related to crop and vegetation richness, respectively, in the local landscape. Differential effects on feeding guilds were also observed in rice-associated communities with the proportional abundance of predators increasing and the proportional abundance of detritivores decreasing with increased landscape rice cover. Thus, we identify a range of rather complex, sometimes contradictory patterns concerning the impact of rice cover and landscape heterogeneity on arthropod community attributes. Importantly, we conclude that that land-use change associated with expansion of monoculture rice need not automatically impact diversity and functioning of the arthropod community.

Knowledge, attitudes and practices of farmers on rodent pests and their management in the lowlands of the Sierra Madre Biodiversity Corridor, Philippines

A survey of the knowledge, attitudes and practices (KAP) of 100 rice farmers and 50 coconut farmers was conducted in the coastal lowland agro-ecosystems of the Sierra Madre Biodiversity Corridor, Luzon, Philippines to identify current rodent management practices and to understand the extent of rat damage and the attitudes of farmers to community actions for rodent management. Pests were most commonly listed as one of the three most important rice and coconut production constraints. Other major crop production constraints were typhoons and insufficient water. Farmers consider rats to be the major pest of coconut and of rice during the wet season rice crop, with average yield losses of 3.0% and 13.2%, respectively. Rice and coconut farmers practised a wide range of rodent management techniques. These included scrub clearance, hunting and trapping. Of the 42 rice farmers and 3 coconut farmers that applied rodenticides to control rodents, all used the acute rodenticide, zinc phosphide. However, only ten rice farmers (23.8%) applied rodenticides prior to the booting stage and only seven farmers (15.6%) conducted pre-baiting before applying zinc phosphide. The majority of farmers belonged to farmer organisations and believed that rat control can only be done by farmers working together. However, during the last cropping season, less than a third of rice farmers (31.2%) applied rodent management as a group. In order to reduce the impact of rodents on the farmers of the coastal lowlands of the Sierra Madre Biodiversity Corridor, integrated management strategies need to be developed that specifically target the pest rodents in a sustainable manner, and community actions for rodent management should be promoted.