Review: Powdery Scab of Potato—Increased Knowledge of Pathogen Biology and Disease Epidemiology for Effective Disease Management

The importance of the potato tuber disease powdery scab, caused by the zoosporic pathogen Spongospora subterranea f. sp. subterranea, has increased worldwide, and the disease is one of the most important problems facing potato production in some regions. This soilborne pathogen produces many resting spores which can remain dormant for long periods, are highly resistant to environmental stresses and can spread the disease on seed potatoes and in contaminated soil. The enigmatic nature of this organism exacerbates the development of effective powdery scab control methods. Substantial knowledge has been gained in the last decade on the biology of the pathogen and the epidemiology of the disease, but no single effective control measure is, or is likely to be, available. An integrated approach to powdery scab management is the aim, with host resistance as a substantial and sustainable component. Further research on the epidemiology of powdery scab and population genetics of the pathogen is urgently required. All stakeholders involved in the potato industry must become aware that solution of the powdery scab problem is likely to be a long-term goal. When resistant cultivars with all the other characteristics demanded for marketing high-quality potatoes are available, and when disease risk from the pathogen on seed and/or in soil can be accurately determined, then the mission to develop effective powdery scab control will become achievable.     

Inheritance of a pectate lyase mediated soft rot resistance in crosses between transgenic potato lines of cv. désirée andS. tuberosum cultivars

Summary Expression of the pectate lyase (PL) isoenzyme 3 in transgenic potato lines of cv. Désirée mediates an enhanced resistance of tuber tissue toErwinia carotovora (Ec) soft rot due to a pre-activation of plant defence mechanisms. Therefore, theSolanum tuberosum cvs Agave and Adretta with a moderate level of soft rot resistance were crossed with such PL-expressing potato lines. The resulting progenies were assessed with respect to plant/tuber characteristics over a period of four years and then tested for PL3-expression as well as for soft rot resistance. 71% of the selected progeny lines exhibited a stable production of the PL3 enzyme. Statistical analysis revealed differences between the transgenic and the non-transgenic progeny concerning the soft rot resistance of tuber tissue. Compared with the PL-inactive progeny, extension of Ec-rotting on the wound surface of PL-transgenic potatoes was diminished on average by 51.8%. Similarly, the degree of cell lysis caused by bacterial maceration was significantly (P<0.05) reduced in tuber tissue of PL-expressing progeny lines. The latter also revealed an enhanced PPO and PAL activity in their tuber tissue indicating an active plant defence. It is concluded therefore that the PL-mediated soft rot resistance introduced into potatoes by means of molecular techniques is heritable.     

Societal Costs of Late Blight in Potato and Prospects of Durable Resistance Through Cisgenic Modification

In the European Union almost 6 Mha of potatoes are grown representing a value of close to €6,000,000,000. Late blight caused by Phytophthora infestans causes annual losses (costs of control and damage) estimated at more than €1,000,000,000. Chemical control is under pressure as late blight becomes increasingly aggressive and there is societal resistance against the use of environmentally unfriendly chemicals. Breeding programmes have not been able to markedly increase the level of resistance of current potato varieties. New scientific approaches may yield genetically modified marker-free potato varieties (either trans- and/or cisgenic, the latter signifying the use of indigenous resistance genes) as improved variants of currently used varieties showing far greater levels of resistance. There are strong scientific investments needed to develop such improved varieties but these varieties will have great economic and environmental impact. Here we present an approach, based on (cisgenic) resistance genes that will enhance the impact. It consists of five themes: the detection of R-genes in the wild potato gene pool and their function related to the various aspects in the infection route and reproduction of the late blight causing pathogen; cloning of natural R-genes and transforming cassettes of single or multiple (cisgenic) R-genes into existing varieties with proven adaptation to improve their value for consumers; selection of true to the wild type and resistant genotypes with similar qualities as the original variety; spatial and temporal resistance management research of late blight of the cisgenic genetically modified (GM) varieties that contain different cassettes of R-genes to avoid breaking of resistance and reduce build-up of epidemics; communication and interaction with all relevant stakeholders in society and transparency in what research is doing. One of the main challenges is to explain the different nature and possible biological improvement and legislative repercussions of cisgenic GM-crops in comparison with transgenic GM-crops. It is important to realize that the present EU Directive 2001/18/EC on GM crops does not make a difference between trans- and cisgenes. These rules were developed when only transgenic GM plants were around. We present a case arguing for an updating and refinement of these rules in order to place cisgenic GM-crops in another class of GM-plants as has been done in the past with (induced) mutation breeding and the use of protoplast fusion between crossable species.     

Applied Biotechnology to Combat Late Blight in Potato Caused by <Emphasis Type="Italic">Phytophthora Infestans</Emphasis>

Potato is an important crop, grown worldwide. It suffers from many pests and diseases among which late blight, caused by the oomycete Phytophthora infestans, is the worst. The disease is still causing major damage in many potato production areas and control is only possible by applying fungicides frequently. The knowledge on the molecular biology and genetics of the interaction between the plant and the oomycete is developing rapidly. These are relevant fields of study, currently dominated by the discovery of many resistance genes and numerous effector proteins and the analysis of their specific mode of action. These studies may yield essential information needed for the development of durable resistance. The long-term and worldwide effort to breed for resistance so far has had little effect. A novel breeding approach may change this. It is based on cisgenic modification (CM) consisting of marker-free pyramiding of several resistance genes and their spatial and temporal deployment yielding dynamic varieties that contain potato genes only. It is envisioned that this CM approach with potato’s own genes will not only prove societally acceptable but may also result in simplifications in the legislation on use of the CM approach. Various parties in the potato research arena intend to cooperate in this novel approach in a number of developing countries where potato substantially contributes to food security. The use of resources such as land, water and energy improves when the effect of late blight is markedly reduced.     

Effect of crop rotation,cultivar and nematicide on growth and yield of potato (Solanum tuberosum L.) in short rotations on a marine clay soil

Summary The effects of cropping frequency on the yield of potato and on the development of soil-borne diseases was studied from 1979 to 1985 in a crop rotation experiment on a marine clay soil. Tuber yield decreased markedly with increasing cropping frequency. The yield of cv. Hertha was reduced by 27% in continuous cropping and by 15% in a wheat/potato or sugar beet/potato rotation, when compared with the rotation wheat/sugar beet/oats/potato. However, a pot experiment showed that yield depression in continuous cropping depended on the cultivar used. Crop growth declined in the second part of the growing season, and senescence accelerated as the cropping frequency increased.Verticillium dahliae was the most important yield-reducing factor. Root infection by this fungus was stimulated by the root-lesion nematodePratylenchus neglectus.     

The current status of knowledge on the cellular biology of potato

Summary Conventional potato breeding does not make full use of the existing biodiversity within theSolanum genus. Moreover breeding at the 4x-4x level is slow. The potential for breeding at the diploid and dihaploid levels has therefore been explored. This requires use of novel techniques to overcome deviations from the desired Endosperm Balance Number. Somatic hybridization approaches include symmetric and asymmetric hybridizations and cybridization. Interesting traits have been successfully transferred through these techniques. Introgression and chromosome elimination have profited from the recent and rapid development of analytical techniques such as RFLP and RAPD. Cellular approaches in potato breeding may be combined with conventional breeding by a stepwise reduction of the ploidy level followed by resynthesis of a new heterozygous tetraploid clone. Such schemes have been used to include virus or nematode resistance. Haploids may be derived from different sources or obtained through different techniques. Dihaploid-dihaploid breeding programmes may be especially interesting. Because of this potential, cellular biology of potato deserves the continued interest of the scientific community.     

Enhanced <Emphasis Type="Italic">Fusarium oxysporum</Emphasis> f. sp. <Emphasis Type="Italic">tuberosi</Emphasis> Resistance in Transgenic Potato Expressing a Rice GLP Superoxide Dismutase Gene

Germin like proteins (GLPs) are a large group of related and ubiquitous plant proteins which are considered to be involved in different processes important for plant development and defense. Multiple functional copies of this gene family have been reported in a number of species (wheat, barley, rice, soybean mosses and liverwort), and their role is being evaluated by gene regulation studies and transgenic approaches. To analyze the role of a rice (Oryza sativa) root expressed germin like protein1 OsRGLP1, for its antifungal activity, transgenic potato plants were developed. These transgenic potato plants were molecularly characterized and biologically assessed after inoculation with Fusarium oxysporum f. sp. tuberosi. Functional analysis showed high accumulation of H₂O₂, increased Superoxide Dismutase (SOD) activity and no oxalate oxidase activity (OxO) in transgenics in comparison to nontransformed control. This increased SOD activity, resistance to heat and sensitivity to H₂O₂ suggest it is a Fe-like SOD. OsRGLP1 expression in potato plants exhibited enhanced resistance in comparison to nontransformed wild type plants suggesting its role in providing protection against Fusarium oxysporum f. sp. tuberosi through elevated SOD level. Overall, results suggest that OsRGLP1 is a candidate for the engineering of potato plants with increased fungal tolerance however, the greater height and tuber number was observed. This phenotype associated with the resistance needs to be evaluated to determine if this is a positive or negative feature.     

Exploring the Natural Biodiversity of Potato for Late Blight Resistance

This paper gives an overview of the activities in Sub-project 4 of the BIOEXPLOIT programme. The European genebank collections of potato need to be narrowed down into (customized) core collections by focusing on loci associated with disease resistance. Therefore, new and more efficient molecular methods are being developed to identify these loci, extensive genotyping of resistance to Phytophthora infestans and signalling loci is being performed, and genotyped selections of accessions are phenotypically analysed for their resistance specificities. The results of these activities are being assembled in an integrated database.     

Variability in the response toPseudomonas solanacearum of transgenic lines of potato carrying a cecropin gene analogue

Summary Transgenic potato plants of cv. Désirée carrying an antibacterial gene, coding for a cecropin lytic peptide analogue, were inoculated with a virulent strain ofPseudomonas solanacearum under controlled conditions. The disease index scored during three repeated infection trials indicated an increased variability in plant response among the transgenic lines which gave either a more susceptible or a more resistant response to the pathogen when compared with untransformed Désirée. Immunity toP. solanacearum was not observed, but it was possible to select a group of transgenic lines that showed resistance levels and disease development curves comparable to the field resistant cv. Cruza 148.     

Resistance levels and fitness of glufosinate-resistant transgenic sweet potato in field experiments

Transgenic herbicide-resistant sweet potato plants [Ipomoea batatas (L.) Lam.] produced through a biolistic transformation were used in this study. The objectives of this research were to (a) quantify resistance levels of transgenic sweet potato expressing the bar gene to glutamine synthetase (GS)-inhibitors, glufosinate and methionine sulfoximine, protoporphyrinogen oxidase (PPO)-inhibiting herbicide, oxyfluorfen and photosynthesis-inhibiting herbicide, paraquat; (b) compare the relative chlorophyll content, quantum yield, growth, and tuberous root yield between transgenic and wild-type sweet potato; (c) compare the response of transgenic and wild-type sweet potato plants to chilling and high temperature, and (d) compare amino acids, minerals and sugar contents of transgenic and wild-type sweet potato tubers. The transgenic sweet potato lines were 20-82-fold more resistant to glufosinate than the wild-type. The representative transgenic line 7171 also showed resistance to methionine sulfoximine, but was not resistant to oxyfluorfen and paraquat, which have different target sites. The stem length, number of leaves, petiole length, shoot fresh weight, and storage roots of transgenic lines were lower than those of the wild-type in field experiments. Reduced growth and root yield were not due to reduced quantum yield and relative chlorophyll contents. Glufosinate increased the ammonium accumulation in leaves of wild-type plants, but had minimal or no effect on leaves of transgenic line 7171. There were differences in mineral contents, free sugars, total amino acids, and free amino acids in the petiole and storage roots between wild-type and line 7171. There was no difference in drought tolerance between wild-type and line 7171, but the transgenic line was more tolerant to chilling temperature than wild-type plants. Follow-up research is required to elucidate the mechanisms underlying the growth and yield differences, different nutritional values, and differential response to low temperature between wild-type and transgenic sweet potato.     

Diploid genotype DW.84-1457, highly resistant to potato leafroll virus (PLRV)

Summary The diploid clone DW.84-1457 which has outstanding resistance to potato leafroll virus (PLRV), has been selected at the Mlochów Centre of the Institute for Potato Research. It has in its pedigree PLRV-resistant clones from the Max Planck Institute nos. MPI 44.1016/10, MPI 44.335/130 and MPI 49.540/2. Its behaviour in the field and response to aphid inoculation indicate high resistance to infection, and the low concentration of the virus in graft-inoculated plants indicates high resistance to multiplication. This combination within one genotype of two aspects of resistance is not connected with hypersensitivity, and is heritable. Clone DW.84-1457 has other desirable characters such as extreme resistance to potato virus X (PVX), high resistance to potato virus M (PVM) and good table and processing quality. It is being utilized in the development of parental lines, both at the diploid and tetraploid level.     

Host genes and transgenes that confer resistance to a Scottish isolate of potato leafroll virus are also effective against a Peruvian isolate

Summary Potato genotypes with host gene-mediated resistance (host-MR) and coat protein-mediated transgenic resistance (CP-MR) to potato leafroll virus (PLRV), were inoculated with a Scottish and a Peruvian isolate of PLRV. The coat protein transgene had been cloned from the Scottish PLRV isolate which had also been used during the screening and selection of genotypes with host resistance. Significantly less PLRV accumulated in plants with either host-MR or CP-MR than in plants of susceptible genotypes or in non-transformed control plants, but the two forms of resistance were equally effective against both PLRV isolates.     

Bacillus thuringiensis cry1Ab gene confers resistance to potato againstHelicoverpa armigera (Hubner)

Summary Helicoverpa armigera is one of the important insect pests adversely affecting the yield of potatoes in India. A synthetic gene encoding the insecticidal crystal protein (Cry1Ab) ofBacillus thuringiensis (Bt) has been introduced into five genotypes of potato usingAgrobacterium tumefaciens. Southern analysis of DNA from transgenic plants confirmed the integration and copy number of the transgene. Double-antibody quantitative sandwich ELISA analysis demonstrated high levels of Cry1Ab protein expression in transgenic plants. Insect bioassays on the leaves of transgenic plants showed considerable protection against the larvae ofH. armigera in terms of leaf area consumed and larval weight reduction.     

马铃薯病毒外壳蛋白融合基因转化马铃薯及其抗病性分析

将多种病毒的有效核酸片断拼接成融合基因转入马铃薯可获得多抗马铃薯材料。针对马铃薯生产中分布广泛、危害严重并经常混合感染的马铃薯X病毒(PVX)、马铃薯Y病毒(PVY)、马铃薯卷叶病毒(PLRV)和马铃薯S病毒(PVS),开展了利用基因工程方法获得兼抗4种马铃薯病毒转基因马铃薯材料的研究。试验在前期获得含4种马铃薯病毒外壳蛋白基因片段的质粒pART27-XSYV-rh的基础上,通过根癌农杆菌(Agrobacterium tumefaciens)介导转化马铃薯(Solanum tuberosum)品种‘陇薯3号’,PCR扩增和PCR-Southern杂交证明,4价融合基因已整合到马铃薯基因组中。qRT-PCR分析表明,该融合基因在转基因植株中能正常表达。3株转基因植株的抗病性鉴定结果表明,2株对4种病毒同时具有抗性;1株对PLRV侵染表现阳性,对另外3种病毒同时具有抗性。     

Production of seed potatoes in Cyprus: the effects of roguing and planting date on the spread of potato leaf roll virus,tuber yield,and infestation by potato tuber moth

Summary Field experiments during 1984–6 tested the effects of planting date on the development of aphid infestations and the spread of potato leaf roll virus (PLRV) in rogued or unrogued plots of potatoes, cv. Spunta. Plantings were made each month from December to April, the customary time for planting being February. Aphid infestation in early-planted plots was severe throughout the growing season; plots planted in February were also severely infested early in the growing season but the populations later gradually declined to undetectable levels. Nevertheless, the incidence of PLRV in the latter plots was as high as in those planted in December-January. Late-planted crops escaped aphid infestation and PLRV infection, either in part (March planting) or completely (April planting). Such crops, however, were uneconomical due to poor yields and heavy losses from potato tuber moth infestation. Roguing significantly reduced the spread of PLRV in all years but its interaction with planting date was insignificant.     

Control of potato blackleg by tuber pasteurisation: the determination of time-temperature combinations for the inactivation of pectolytic erwinia

Summary Pasteurisation of potato tubers by immersion in hot water is being examined as a method of blackleg control. Studies on the in vitro heat resistance of representative pectolytic erwinia have enabled prediction curves for the selection of suitable times and temperatures of pasteurisation to be prepared. High levels of contamination on some tubers may require heat treatment that will reduce the erwinia population by more than 80% to ensure that only innocuous levels remain.     

Biotechnology initiative to achieve plant pest and disease resistance

Biotechnology applications are primarily proprietary, with 70% of the annual global research expenditures of US$11 × 10⁹ being in the private sector. The International Service for the Acquisition of Agri-Biotech Applications (ISAAA) addresses this challenge by building a new partnership of three donor groups (bilateral/multilateral agencies, private philanthropic foundations, and private companies) to support a not-for-profit institution within existing centres of excellence (Cornell University in North America, John Innes Centre in Europe, Technova in the Asian Pacific Rim, and three additional locations to be established in developing countries). Brokered projects involve only near-term technologies with a high probability of success, using tissue culture/micropropagation, disease diagnostics and transgenic plants. The main targets for transgenic plants are coat protein-mediated protection for non-conventional virus resistance (NCVR) and insecticidal proteins from Bacillus thuringiensis (Bt); both NCVR and Bt can contribute to integrated pest management strategies using reduced inputs for chemical control of insects. Four ISAAA projects demonstrate the potential of NCVR and Bt in developing countries: (1) potato resistant to potato viruses X and Y with Mexico/Monsanto/Rockefeller Foundation, (2) melon resistant to cucumber mosaic virus with Costa Rica/Asgrow Seed/USAID and the ISAAA Fellowship Program, (3) papaya resistant to papaya ringspot virus with Brazil and other countries/Cornell University/Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA), and (4) cotton resistant to Lepidoptera with Brazil/EMBRAPA. Although over 1000 field trials of transgenic plants have taken place worldwide, those experiments have been mostly small scale and do not provide sufficient data to assess either the durability of resistance achieved with genetic engineering or conclusive public acceptance of this technology. However, resistance management modelling is speculative and can not decisively indicate a deployment strategy in the absence of continued monitoring and experimentation. ISAAA's projects should allow opportunities for assessment of resistance management practices for applied plant biotechnology to control agricultural pests under diverse production systems.