Molecular,morphological and agronomic characterization of the sweet potato (Ipomoea batatas L.) germplasm collection from Mozambique: Genotype selection for drought prone regions

Sweet potato (Ipomoea batatas (L.) Lam) is one of the most important root crops in Mozambique, ranking in the 3rd position, after cassava and maize. Within the scope of the national and regional strategies/initiatives, we have used a multi-analysis approach to characterize the national sweet potato germplasm collection at two different levels: i) genetic, morphological and agronomic diversity; and ii) agronomic potential (storage root yield, vine weight, biomass, harvest index and dry matter content) toward drought tolerance. This collection, composed by 44 accessions, comprises 28 genotypes cultivated in three different provinces of Mozambique (Gaza, Inhambane and Zambezia), nine from other African countries (Kenya, South Africa, Uganda and Zimbabwe), one from the United States of America, and six from CGIAR research centers (IITA and CIP). According to our results, the Mozambican germplasm bank presents a high level of diversity, comparable to those from the collections of the primary centers of origin and South Africa, therefore constituting of a good source of agronomic traits for breeding. Regarding drought tolerance, six Mozambican genotypes (Admarc, Chingova, Nhacoongo-1, Xihetamakote, Nwanatuyo, and Chissicuana-2), one from Uganda (NASPOT-5), one from Zimbabwe (Moz_white), one from Kenya (SPK 004), and one from the USA (Resisto) seem to have the highest potential to be used in regions with frequent drought seasons and in future breeding programs. The results showed that such integrated analysis can be used to successfully characterize the genetic material in terms of suitability to drought-prone regions, therefore helping sweet potato crop management, with economic and food security impacts.     

Resistance to sweet potato virus disease (SPVD) in wild East African Ipomoea

Sweet potato virus disease (SPVD), the most harmful disease of sweet potatoes in East Africa, is caused by mixed infection with sweet potato feathery mottle potyvirus (SPFMV) and sweet potato chlorotic stunt crinivirus (SPCSV). Wild Ipomoea spp. native to East Africa (J cairica, I. hildebrandtii, I. involucra and J wightii) were graft-inoculated with SPVD-affected sweet potato scions. Inoculated plants were monitored for symptom development and tested for SPFMV and SPCSV by grafting to the indicator plant J setosa, and by enzyme-linked immunosorbent assay (ELISA). Virus-free scions of sweet potato cv. Jersey were grafted onto these wild Ipomoea spp. in the field, and scions collected 3 wk later were rooted in the greenhouse and tested for viruses using serological tests and bioassays. In all virus tests, J cairica and J involucra were not infected with either SPFMV or SPCSV. J wightii was infected with SPFMV, but not SPCSV, in the field and following experimental inoculation; J hildebrandtii was infected with SPCSV, but not SPFMV, following experimental inoculation. These data provide the first evidence of East African wild Ipomoea germplasm resistant to the viruses causing SPVD.     

Detection,identification and differentiation of Pectobacterium and Dickeya species causing potato blackleg and tuber soft rot: a review

The soft rot Enterobacteriaceae (SRE) Pectobacterium and Dickeya species (formerly classified as pectinolytic Erwinia spp.) cause important diseases on potato and other arable and horticultural crops. They may affect the growing potato plant causing blackleg and are responsible for tuber soft rot in storage thereby reducing yield and quality. Efficient and cost‐effective detection and identification methods are essential to investigate the ecology and pathogenesis of the SRE as well as in seed certification programmes. The aim of this review was to collect all existing information on methods available for SRE detection. The review reports on the sampling and preparation of plant material for testing and on over thirty methods to detect, identify and differentiate the soft rot and blackleg causing bacteria to species and subspecies level. These include methods based on biochemical characters, serology, molecular techniques which rely on DNA sequence amplification as well as several less‐investigated ones.     

Reproductive incompatibility and cytochrome oxidase I gene sequence variability amongst host-adapted and geographically separate Bemisia tabaci populations (Hemiptera: Aleyrodidae)

Abstract. Reciprocal‐crossing experiments were carried out and mitochondrial cytochrome oxidase I gene (mtCOI) sequences were compared for allopatric and sympatric Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) populations collected from Africa and India, and from the host‐plants cassava, sweet‐potato and a common weed, Euphorbia geniculata. Three incompatible mating groups were discovered, which involved the cassava B. tabaci colonies from Africa and India, the cassava and sweet‐potato B. tabaci populations from Uganda, and the cassava and E. geniculata B. tabaci from India. Successful reciprocal mating occurred between cassava‐specific B. tabaci from Uganda, Tanzania and Ghana, and between two Indian cassava B. tabaci populations. The parsimony and neighbour‐joining analyses of 699 bp mtCOI gene sequences divided the colonies primarily into those originating from Africa and India. Further subgrouping corresponded to host‐plant specialization. Cassava‐specific Ugandan, Tanzanian and Ghanaian colonies formed a single group and the sympatric sweet‐potato colony from Uganda grouped separately from them. The two geographically distant Indian cassava B. tabaci populations were similar and formed a single group, whereas the sympatric E. geniculata colony formed a sister clade. The clades generated by the phylogenetic analyses were maintained, with highly supported bootstrap values, when other published mtCOI gene sequences were included in the tree‐building process and the divisions matched those revealed by the reciprocal‐crossing experiments. These data suggest that biologically discrete populations exist within B. tabaci (sensu Russell, 1957 ).     

Gene-based microsatellites for cassava (Manihot esculenta Crantz): prevalence, polymorphisms, and cross-taxa utility

Background  

Cassava (Manihot esculenta Crantz), a starchy root crop grown in tropical and subtropical climates, is the sixth most important crop in the world after wheat, rice, maize, potato and barley. The repertoire of simple sequence repeat (SSR) markers for cassava is limited and warrants a need for a larger number of polymorphic SSRs for germplasm characterization and breeding applications.     

A sucrose non‐fermenting‐1‐related protein kinase‐1 gene,IbSnRK1, improves starch content,composition, granule size,degree of crystallinity and gelatinization in transgenic sweet potato

Sucrose non‐fermenting‐1‐related protein kinase‐1 (SnRK1) is an essential energy‐sensing regulator and plays a key role in the global control of carbohydrate metabolism. The SnRK1 gene has been found to increase starch accumulation in several plant species. However, its roles in improving starch quality have not been reported to date. In this study, we found that the IbSnRK1 gene was highly expressed in the storage roots of sweet potato and strongly induced by exogenous sucrose. Its expression followed the circandian rhythm. Its overexpression not only increased starch content, but also decreased proportion of amylose, enlarged granule size and improved degree of crystallinity and gelatinization in transgenic sweet potato, which revealed, for the first time, the important roles of SnRK1 in improving starch quality of plants. The genes involved in starch biosynthesis pathway were systematically up‐regulated, and the content of ADP‐glucose as an important precursor for starch biosynthesis and the activities of key enzymes were significantly increased in transgenic sweet potato. These findings indicate that IbSnRK1 improves starch content and quality through systematical up‐regulation of the genes and the increase in key enzyme activities involved in starch biosynthesis pathway in transgenic sweet potato. This gene has the potential to improve starch content and quality in sweet potato and other plants.     

Effects of potnetial trap crops and planting date on soil infestation with potato cyst nematodes and root-knot nematodes

In 1997 and 1998 the stimulation of hatch of potato cyst nematodes (PCN) by a trap crop was studied at various times during the growing season in a container and a field experiment. Solanum nigrum‘90‐4750‐188’was used as the trap crop in both experiments and was sown on 1 May, 16 June or 1 August in two successive years on different plots. Neither experiment revealed much seasonal variation in hatchability of PCN juveniles under a trap crop. In the container experiment, the hatch of the Globodera pallida Pa3 population was equally and strongly stimulated (89%) at all sowing dates in both years, except for the 1 August sowing in 1998 (when the hatch was 77% under extremely wet soil conditions). In the control treatment with non‐hosts (flax followed by barley) the total spontaneous hatch was 50% over 2 yr. In the field experiment, the hatch of PCN, averaged over the four populations, was also equally stimulated (71%) at all sowing dates in both years. In the control treatment with non‐hosts (flax‐barley) the total spontaneous hatch was 36% over 2 yr. Total hatch under the trap crop over 2 yr varied between the four PCN populations from 63% to 80%. In 1998 and 1999, control of potato cyst nematodes (PCN) by the potential trap crops Solanum sisymbriifolium and S. nigrum‘90‐4750‐188’was studied in the field. Potato was also included as a trap crop. In the 1998 experiment, potato, S. sisymbriifolium and S. nigrum strongly stimulated the hatch of PCN compared with the non‐host white mustard (Sinapis alba). Roots of potato and white mustard were mainly found in the top 10 cm of soil, whereas roots of S. sisymbriifolium and S. nigrum were also abundant at depths of 10–20 cm and 20–30 cm. In the 1999 experiment, soil infestation with PCN decreased markedly with potato and S. sisymbriifolium as trap crops. In plots moderately to severely infested with 2‐yr old cysts (2–29 juveniles ml^?1 air dried soil), potato reduced soil infestation by 87% and S. sisymbriifolium by 77%. In plots moderately to severely infested with 1‐yr old cysts the reductions were 74% and 60%, respectively. The reduction was least on plots very severely infested with PCN (110–242 juveniles ml^?1 soil): 69% and 52% for potato and S. sisymbriifolium, respectively. Soil infestations of plots that were initially slightly to severely infested with the root‐knot nematode Meloidogyne hapla were greatly reduced under fallow and S. sisymbriifolium but increased under potato. From these and previous experiments it was concluded that, for several reasons, S. sisymbriifolium is a promising trap crop.     

Tuber production,dormancy and resistance against Phthorimaea operculella (Zeller) in wild potato species

The diversification of resistant potato varieties at a landscape level could slow adaptation by Phthorimaea operculella to potato resistance and promote sustainable crop protection. In this study, we assessed wild potato species as novel sources of foliage and tuber resistance against P. operculella. Tuber resistance was quantified for 136 and foliage resistance for 54 potato accessions representing 14 and nine potato species, respectively. Several accessions were highly resistant to moth damage in tubers and/or foliage. In particular, Solanum chiquidenum and Solanum sandemanii were highly resistant to damage in tubers. Several accessions of Solanum multiinterruptum and a small number of accessions of Solanum bukasovii, Solanum berthaultii, Solanum sparsipilum and Solanum wittmackii also had highly resistant tubers. Larval survival on foliage of S. bukasovii and S. chiquidenum was generally low. New resistance sources are listed, and insect performance on the plants is described with possible resistance mechanisms. The study also examined potential trade‐offs associated with resistance. Tuber resistance was negatively correlated with the number and weight of tubers produced per plant, but positively correlated with the length of dormancy across accessions, indicating that, although long dormancy is not a prerequisite for resistance, species and accessions with extended dormancy will have more resistant tubers. Tuber and foliage resistance were generally positively correlated across all accessions; however, among accessions from within a potato species, there were negative (S. berthaultii), positive (S. chiquidenum) and non‐significant (S. bukasovii) relations. These results indicate that, besides identifying novel resistance sources, an improved understanding of the mechanisms and inherent trade‐offs associated with tuber and foliage resistance will improve the efficiency of potato breeding programmes aimed at enhancing resistance against P. operculella.     

Ex situ conservation of plant germplasm using biotechnology

Conservation of plant genetic resources attracts more and more public interest as the only way to guarantee adequate food supplies for future human generations. However, the conservation and subsequent use of such resources are complicated by cultural, economical, technical and political issues. Over the last 30 years, there have been significant increases in the number of plant collections and in accessions in ex situ storage centres throughout the World. The present review is of these ex situ collections and the contribution biotechnology has made and can make to conservation of plant germplasm. The applications and limitations of the new, molecular approaches to germplasm characterization are discussed. In vitro slow growth is used routinely for conserving germplasm of plants such as banana, plantain, cassava and potato. More recently, cryopreservation procedures have become more accessible for long-term storage. New cryopreservation techniques, such as encapsulation-dehydration, vitrification and desiccation, lengthen the list of plant species that can not only tolerate low temperatures but also give normal growth on recovery. Extensive research is still needed if these techniques are to be fully exploited.V.M. Villalobos is with the Food and Agriculture Organization of the United Nations, Viale delle Terme di Caracalia, 00100 Rome, Italy. F. Engelmann is with the International Plant Genetic Resources Institute (IPGRI), Via delle Sette Chiese 142, 00145 Rome, Italy.     

Effect of the trap crop Solanum sisymbriifolium and two biocontrol fungi on reproduction of the potato cyst nematode,Globodera pallida

The potato cyst nematode, Globodera pallida, is one of the most important pests of potato worldwide. Owing to regulatory considerations and potential environmental impact, control options for this nematode are becoming increasingly limited. Solanum sisymbriifolium and biological control agents offer viable alternative options for controlling G. pallida. Therefore, experiments were conducted to determine the effect of the nematode trap crop S. sisymbriifolium, alone or in combination with the biocontrol agents Trichoderma harzianum or Plectosphaerella cucumerina, on population decline of G. pallida. Experiments were conducted for three different ‘cropping systems’: potato (Solanum tuberosum), S. sisymbriifolium, or soil only (fallow), each followed by a potato crop. Soil was amended with P. cucumerina, T. harzianum or left unamended, and then infested with nematodes at a rate of five eggs g^?1 of soil. After 16 weeks in the greenhouse, plants were removed and the soil containing cysts was refrigerated at 4°C for 8 weeks, and then planted to potato. Cysts of G. pallida were counted after an additional 16‐week period. The P_f/P_i of G. pallida was significantly reduced by 99% in potato following S. sisymbriifolium compared to both the potato‐following‐fallow and the potato‐following‐potato treatments. Amendment of soil with T. harzianum significantly reduced P_f/P_i of G. pallida by 42–47% in the potato‐following‐potato but not in either the potato‐after‐fallow nor in the potato‐after‐S. sisymbriifolium cycles which supports evidence that the plant species may play a role in the biocontrol activity of this fungus. Addition of the fungus P. cucumerina resulted in a 64% decrease in P_f/P_i in the potato‐following‐fallow in one experiment, and an 88% decrease in P_f/P_i in potato‐following‐potato but the decrease in P_f/P_i was not consistent over all experiments. However, both biocontrol fungi resulted in lower numbers of progeny cysts after an initial 16‐week incubation with potato. To look at the effect of varied population density of the nematode on efficacy of S. sisymbriifolium to reduce G. pallida populations, potato, S. sisymbriifolium, or barley were planted into soil infested with G. pallida at rates of 5, 20 or 40 eggs g^?1 soil applied as cysts (20, 80 or 160 cysts pot^?1). After 16 weeks, numbers of cysts produced in each treatment were determined for each infestation rate. No new cysts were recovered from either S. sisymbriifolium or barley treatments, confirming that neither plant is a host for G. pallida. High numbers of cysts were recovered with potato. Soil from each treatment (containing original cysts and newly‐formed cysts when present) were then planted with potato. After an additional 16 weeks, few cysts were found in the potato‐after‐ S. sisymbriifolium treatments regardless of initial infestation rate. When potato followed barley, numbers of cysts were similar to those found after a single cycle of potato, indicating that the barley crop had no effect on the survival of initial inoculum. Overall, these results suggest that S. sisymbriifolium has potential to significantly reduce G. pallida populations, and also that the cropping system (i.e. the sequence of non‐host and host plants) may play a significant role in the efficacy of fungal biological control agents.     

Sweet potato (Ipomoea batatas) trypsin inhibitors expressed in transgenic tobacco plants confer resistance against Spodoptera litura

A sweet potato (Ipomoea batatas cv. Tainong 57) trypsin inhibitor gene was introduced into tobacco plants (Nicotiana tabaccum cv. W38) by Agrobacterium tumefaciens– mediated transformation. From 30 independent transformants, three lines with high level of expression were further analyzed. The trypsin inhibitor gene, under control of the 35S CaMV promoter, led to the production of the trypsin inhibitor proteins up to 0.2% of the total protein. In insecticidal bioassays of transgenic tobacco plants, larval, growth of Spodoptera litura (F.), the tobacco cutworm, was severely retarded as compared to their growth on control plants. This observation implied that expression of sweet potato trypsin inhibitor can provide an efficient method for crop protection. Received: 29 July 1996 / Revision received: 15 November 1996 / Accepted: 8 December 1996     

Synthesis of a ricin toxin B subunit-rotavirus VP7 fusion protein in potato

A gene encoding the outer capsid glycoprotein (VP7) of simian rotavirus SA11, was genetically linked to the amino terminus of the ricin toxin B subunit (RTB) isolated from castor-oil plant (Ricinus communis) seeds. To assess fusion protein expression in plant cells, the VP7::RTB fussion gene was transferred into potato (Solanum tuberosum) cells by Agrobacterium tumefaciens-mediated transformation methods and transformed plants regenerated. The fusion gene was detected in transformed potato genomic DNA by polymerase chain reaction DNA amplification methods. Immunoblot analysis with anti-SA11 antiserum as the primary antibody verified the presence of VP7::RTB fusion protein in transformed potato tuber tissues. The plant-synthesized fusion protein bound RTB membrane receptors as measured by asialofetuin-enzyme-linked immunosorbent assay (ELISA). The ELISA results indicated that the VP7::RTB fusion protein was biologically active and made up approx 0.03% of total soluble transformed tuber protein. The biosynthesis of receptor binding VP7::RTB fusion protein in potato tissues demonstrates the feasibility of producing monomeric ricin toxin B subunit adjuvant-virus antigen fusion proteins in crop plants for enhanced immunity.     

A Multiplex Microsatellite Marker Kit for Diversity Assessment of Large Cassava (<Emphasis Type="Italic">Manihot esculenta</Emphasis> Crantz) Germplasm Collections

Current methods for molecular fingerprinting of cassava (Manihot esculenta Crantz) have limited throughput or are costly, thus preventing the characterization of large germplasm collections such as those held by the International Agricultural Research Centers or National Research Institutions, which comprise hundreds to thousands of accessions. Here, we report the development of a fluorescence-based multiplex simple sequence repeat (SSR) marker kit that enables accurate and cost-effective cassava fingerprinting. The kit comprises 16 SSR markers assembled into five multiplex panels and was tested on 21 cassava cultivars alongside one accession of Manihot epruinosa, a wild relative. A total of 68 alleles were detected with, on average, 4.25 alleles per locus and a polymorphism information content of 0.53. The marker kit reported here is comparable to previously published amplified fragment length polymorphism and SSR marker systems in terms of discriminating power and informativeness while offering significant advantages in speed and cost of marker analysis. Previous molecular genetic diversity studies have suggested that cassava germplasm collections contain duplicate entries based on the occurrence of identical genetic profiles. Using the newly developed microsatellite kit, three out of six putative duplicate accessions could be readily differentiated, showing that these are distinct genotypes. The relevance of these findings with respect to the characterization and management of large cassava germplasm collections is discussed.     

<Emphasis Type="BoldItalic">In vitro</Emphasis> availability of some essential minerals in commonly eaten processed and unprocessed Caribbean tuber crops

The levels of three essential minerals Ca, Fe and Mg and the extent of their availability were assessed in four commonly eaten Caribbean tuber crops [dasheen (Xanthosoma spp.), Irish potato (Solanum tuberosum), sweet potato (Ipomoea batatas) and yellow yam (Dioscorea cayenensis)] in their processed and unprocessed states. Calcium was highest in cooked dasheen (5150±50 mg/kg) while Magnesium was highest in uncooked Irish potato (3600±200 mg/kg). There was no significant loss of calcium from the food samples upon cooking. All the uncooked food samples displayed higher levels minerals assessed compared to the cooked samples except for cooked Irish potato that recorded the level of iron (182.25±8.75 mg/kg). Availability of these minerals in the cooked and uncooked tubers crops upon digestion also showed a similar pattern. In conclusion, the consumption of these tuber crops in the Caribbean may not be responsible for the reported cases of iron deficiency in the region. However, the␣availability of minerals from these tuber crops when consumed with other foods (the usual practice in the Caribbean) needs further investigation.     

Assessing genetic potential in germplasm collections of crop plants by marker-trait association: a case study for potatoes with quantitative variation of resistance to late blight and maturity type

Genetic diversity of crop plants resulting from breeding and selection is preserved in gene banks. Utilization of such materials for further crop improvement depends on knowledge of agronomic performance and useful traits, which is usually obtained by phenotypic evaluation. Associations between DNA markers and agronomic characters in collections of crop plants would (i) allow assessment of the genetic potential of specific genotypes prior to phenotypic evaluation, (ii) identify superior trait alleles in germplasm collections, (iii) facilitate high resolution QTL mapping and (iv) validate candidate genes responsible for quantitative agronomic characters. The feasibility of association mapping was tested in a gene bank collection of 600 potato cultivars bred between 1850 and 1990 in different countries. The cultivars were genotyped with five DNA markers linked to previously mapped QTL for resistance to late blight and plant maturity. Specific DNA fragments were tested for association with these quantitative characters based on passport evaluation data. Highly significant association with QTL for resistance to late blight and plant maturity was detected with PCR markers specific for R1, a major gene for resistance to late blight, and anonymous PCR markers flanking the R1 locus at 0.2 Centimorgan genetic distance. The marker alleles associated with increased resistance and later plant maturity were traced to an introgression from the wild species S. demissum. These DNA markers are the first marker that are diagnostic for quantitative agronomic characters in a large collection of cultivars.     

过表达IbOr基因甘薯增强抗旱性的生理机制

以超表达甘薯橙色基因(IbOr)的转基因甘薯(TS)以及非转基因甘薯(NT)为实验材料,通过15%聚乙二醇6000(PEG-6000)模拟干旱条件,研究转基因与非转基因甘薯幼苗在水分胁迫不同时间的光合系统,膜脂过氧化及抗氧化防御系统中主要指标的变化情况,探讨转基因甘薯耐旱性的生理机制。结果显示:(1)随PEG-6000胁迫时间延长,甘薯叶片的叶绿素、类胡萝卜素含量及其叶片净光合速率、气孔导度、胞间CO2浓度、蒸腾速率都显著降低,但转基因株系降低幅度小于非转基因植株。(2)在正常供水和水分胁迫下,超表达IbOr基因甘薯叶片中O-·2、MDA含量均低于非转基因甘薯,即转基因甘薯具有较低的活性氧水平且脂膜受损伤较小。(3)PEG-6000胁迫24h后,甘薯叶片中SOD、POD酶活性均增加,48h达到最大值,且转基因甘薯中2种酶活性显著高于非转基因甘薯。研究表明,过表达IbOr基因可以有效减轻甘薯在水分胁迫条件下受损害的程度,且可能主要通过提高甘薯的抗氧化胁迫能力来完成。     

湖南甘薯种质资源曲叶病毒检测分析初报

本研究依托"第三次全国农作物种质资源普查与收集行动",利用巢式PCR(Nested PCR)检测技术,对从湖南各地区采集的甘薯种质资源进行甘薯曲叶病毒的调查、检测、统计与分析,获得该地区甘薯种质资源曲叶病毒的感染和分布情况。对收集的246份甘薯种质资源进行了甘薯曲叶病毒病症状的调查,记录了每份种质资源的田间生长特性;建立了一种甘薯曲叶病毒巢式PCR检测技术,该技术相对其他技术具有特异性强、灵敏度高、检测通量大、检测成本低的特点。利用建立的巢式PCR检测技术对选取的样品进行甘薯曲叶病毒检测,分析检测结果发现:(1)巢氏PCR共检测出14份甘薯种质资源感染甘薯曲叶病毒,根据病毒基因测序结果分析湖南省至少存在2种曲叶病毒株系。(2)田间调查共发现8份甘薯种质资源的叶片具有甘薯曲叶病毒病典型的卷曲症状,但是其中仅有4份资源与曲叶病毒巢氏PCR检测结果一致;另外4份资源虽然具有明显的卷叶现象但是未检测出曲叶病毒。(3)曲叶病毒检测呈阳性的14份甘薯种质资源分别来源于邵阳市、长沙市、永州市和株洲市4个地区,占种质资源总数的5.7%;4个地区甘薯种质资源的病毒感染率分别为17.6%、14.5%、7.1%和6.7%;全省范围内的种质资源染病情况具有较大的地域差异性;综合甘薯种植情况和地理环境分析,商品薯的跨区域流通和农民自留种的种植习惯是影响甘薯病毒传播的主要因素。本研究首次利用巢式PCR技术对湖南地区甘薯曲叶病毒进行检测和调查,为甘薯种质资源的保存、繁殖、鉴定与利用提供了重要的技术支撑,也为湖南地区甘薯曲叶病毒侵染情况及相关的分子生物学研究提供了数据参考。     

Large-scale genome-wide association study,using historical data,identifies conserved genetic architecture of cyanogenic glucoside content in cassava (Manihot esculenta Crantz) root

Manihot esculenta (cassava) is a root crop originating from South America that is a major staple in the tropics, including in marginal environments. This study focused on South American and African germplasm and investigated the genetic architecture of hydrogen cyanide (HCN), a major component of root quality. HCN, representing total cyanogenic glucosides, is a plant defense component against herbivory but is also toxic for human consumption. We genotyped 3354 landraces and modern breeding lines originating from 26 Brazilian states and 1389 individuals were phenotypically characterized across multi-year trials for HCN. All plant material was subjected to high-density genotyping using genotyping by sequencing. We performed genome-wide association mapping to characterize the genetic architecture and gene mapping of HCN. Field experiments revealed strong broad- and narrow-sense trait heritability (0.82 and 0.41, respectively). Two major loci were identified, encoding for an ATPase and a MATE protein, and contributing up to 7 and 30% of the HCN concentration in roots, respectively. We developed diagnostic markers for breeding applications, validated trait architecture consistency in African germplasm and investigated further evidence for the domestication of sweet and bitter cassava. Fine genomic characterization revealed: (i) the major role played by vacuolar transporters in regulating HCN content; (ii) the co-domestication of sweet and bitter cassava major alleles are dependent upon geographical zone; and (iii) the major loci allele for high HCN in M. esculenta Crantz seems to originate from its ancestor, M. esculenta subsp. flabellifolia. Taken together, these findings expand our insights into cyanogenic glucosides in cassava roots and its glycosylated derivatives in plants.     

Breaking the Cycle of Xanthomonas campestris pv. musacearum in Infected Fields through the Cultivation of Annual Crops and Disease Control in Adjacent Fields

Cultivation of non‐host crops after uprooting Xanthomonas campestris pv. musacearum (Xcm)‐infected banana plants has been advocated for breaking Xanthomonas wilt disease (XW) cycle in fields. Knowledge on the interaction of these crops with Xcm is limited. Maize, beans and sweet potato were planted after uprooting Xcm‐infected banana plants in Rwanda and eastern Democratic Republic of Congo (DR Congo). A weed fallow (mixed species) served as the control. After one, two and/or four break‐crop or fallow seasons, healthy plantlets were replanted and monitored for XW for 12–24 months. XW status in adjacent fields was monitored, and diseased stems within 100–300 m radius of the two‐ and four‐season experiments were uprooted. In Rwanda, soil and plant parts from the one‐season experiments were sampled for Xcm isolation and Xcm‐like colonies confirmed with Xcm‐specific primers using PCR. Pathogenicity tests were performed to confirm the ability of the PCR‐positive isolates to infect healthy banana plantlets. XW was observed in all the one‐season experiments, with higher cumulative incidences in maize and bean plots. However, no similar trends were observed in the two‐season experiments, with a 6–8% incidence observed only in bean and potato plots in DR Congo. Lengthening time under break crops to two and four seasons, respectively, reduced the incidence to 3% and zero in Rwanda and 0–8% in the two‐season experiments in DR Congo. Incidence in the first‐season experiments highly correlated (R = 88) to that in the adjacent fields, suggesting possible re‐infections from these fields. Two season with break crop plus collective XW control are recommended in these agro‐ecosystems. PCR‐positive Xcm‐like colonies from break crops only induced localized cell death on banana, while PCR‐positive isolates from symptomatic banana plants caused full XW symptoms. Cross‐infection/inoculation studies under controlled conditions are still needed to conclusively elucidate Xcm interaction with these crops.