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.     

Yield Response of Potato (<Emphasis Type="Italic">Solanum tuberosum L</Emphasis>.) Genotypes to Late Blight Caused by <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Uganda

Potato (Solanum tuberosum L.) is a major food and cash crop, mainly grown by small-scale farmers in the highland regions of Uganda. Potato late blight is one of the major diseases limiting production with potential yield losses over 70%, making host resistance a strong element in integrated disease management. This study was carried out to screen and select high yielding potato genotypes with resistance to late blight in Uganda. Forty-eight genotypes, including advanced clones from the population B3C2 of the International Potato Centre, commercial and farmers’ varieties, were evaluated under two environments for two seasons. Trials were laid out in an 8?×?6 alpha lattice design with three replications. Genotypes showed significant differences in yield and resistance to blight. A higher disease severity was observed in Karengyere (56%). The average RAUDPC (= 100 max) across locations indicated that genotypes 395,077.12 and 392,657.8, with disease severity of 12% and 14%, respectively, were the most resistant. Genotypes Victoria (53%) and NKRN59.124 (48%) were the most susceptible. Mean tuber yield under late blight infection was19.8 t ha^?1. The best yielding genotype across sites was 395,112.32 (35.6 t ha^?1) while 394,905.8 (10.3 t ha^?1), yielded the lowest. The mean marketable tuber weight was 8.9 kg with genotypes 395,112.32 and 395,109.34 having the highest marketable weight of 16.5 kg and 15.6 kg respectively. Correlations between yield and yield related parameters were positive (p ≤?0.001), while those between RAUDPC were negative. The following genotypes, 395,112.32, 391,919.3, 393,220.54. 393,077.54, 396,038.107. 392,657.8, Kinigi, 395,014.17, NKRN59.58, NKRK19.17 and 395,011.2, were identified as promising parents for a late blight resistance breeding program. These exhibited high to medium resistance to late blight disease and high yields.     

Amplification of the <Emphasis Type="Italic">Phytophthora infestans</Emphasis> RG57 loci Facilitates <Emphasis Type="Italic">in planta</Emphasis> T-RFLP Identification of Late Blight Genotypes

Late blight, caused by the oomycete Phytophthora infestans (Mont.) de Bary, is a devastating disease in potato and tomato and causes yield and quality losses worldwide. The disease first emerged in central America and has since spread in North America including the United States and Canada. Several new genotypes of P. infestans have recently emerged, including US-22, US-23 and US-24. Due to significant economic and environmental impacts, there has been an increasing interest in the rapid identification of P. infestans genotypes. In addition to providing details regarding the various phenotypic characteristics such as fungicide resistance, host preference, and pathogenicity associated with various P. infestans genotypes, information related to pathogen movement and potential recombination may also be determined from the genetic analyses. Restriction fragment length polymorphism (RFLP) analysis with the RG57 loci is one of the most reliable procedures used to genotype P. infestans. However, the RFLP procedure requires propagation and isolation of the pathogen and relatively large amounts of DNA. Isolation of the late blight pathogen is sometimes impossible due to the poor condition of the infected tissues or the presence of fungicide residues. In this study, we describe a procedure to identify P. infestans at the molecular level in planta using terminal restriction fragment length polymorphism (T-RFLP) of the RG57 loci. This T-RFLP assay is sufficiently sensitive to detect and differentiate P. infestans genotypes directly in planta without propagation and isolation of the pathogen, to facilitate the timely implementation of best management practices.     

Effects of Mulch and Potato Hilling on Development of Foliar Blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) and the Control of Tuber Blight Infection

Foliar and tuber blight caused by Phytophthora infestans accounts for significant losses in potatoes in field and storage. Nevertheless, limited research has been published on the effects of cultural practices on late blight control. Field experiments were conducted in two years on Howard gravely loam soil in New York State to evaluate the effectiveness of mulching using oat straw and hilling in preventing tuber blight infection for cvs Allegany and Katahdin. Potato hilling and mulching had little effect on foliar blight development. The cultivar affected the disease development in the foliage, with cv. Allegany showing lower foliar late blight than cv. Katahdin. Tuber blight incidence averaged 25% for cv. Allegany and 3% for cv. Katahdin in hilled plots, while in the mulched plots the incidence of tuber blight averaged 33% for cv. Allegany and 10% for cv. Katahdin. The straw hay mulch was ineffective in tuber blight control. Tubers set at a soil depth of more than 7 cm had lower tuber blight incidence than shallow tubers set at a depth of less than 7 cm. In both years, hilling provided partial protection of tubers but its effectiveness was limited in the presence of favourable conditions for late blight development. Even though large hills had proportionally a lower tuber blight incidence than medium-sized hills, the difference between the different hill sizes was not significant. These studies suggest that the use of cultivars with foliage resistance to late blight in combination with cultural practices may partially reduce the incidence of tuber blight. Mention of trade names or commercial products in this publication is solely for the purpose of providing specific information and does not imply recommendation for endorsement by Cornell University or the US Department of Agriculture.     

Progress in Mapping and Cloning Qualitative and Quantitative Resistance Against <Emphasis Type="Italic">Phytophthora infestans</Emphasis> in Potato and Its Wild Relatives

Cultivated potato is susceptible to many pests and pathogens, none of which is more of a threat to potato agriculture than the late blight disease, caused by the oomycete Phytophthora infestans (Mont.) de Bary. To date all efforts to thwart this most adaptive of pathogens have failed, and early attempts to deploy ‘R genes’ introgressed from the wild Mexican hexaploid Solanum demissum ended in abject failure. With the advent of facile gene mapping and cloning, allied to knowledge of plant resistance gene structure, renewed efforts are leading to mapping and isolation of new sources of late blight resistance in potato wild species, many of which are being performed under the auspices of the BIOEXPLOIT project (Sub-project 2). We document recent advances in late blight resistance gene mapping and isolation, and postulate how these genes, allied to knowledge of pathogen effectors and their recognition specificity, may greatly enhance our chances of halting the progress of late blight disease in potato crops worldwide.     

Maturity-Adjusted Resistance of Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Cultivars to Verticillium Wilt Caused by <Emphasis Type="Italic">Verticillium dahliae</Emphasis>

Verticillium wilt is a fungal disease of potato caused by two species of Verticillium, V. dahliae and V. albo atrum. The pathogen infects the vascular tissue of potato plants through roots, interfering with the transport of water and nutrition, and reducing both the yield and quality of tubers. We have evaluated the reaction of 283 potato clones (274 cultivars and nine breeding selections) to inoculation with V. dahliae under greenhouse conditions. A significant linear correlation (r = 0.4, p < 0.0001) was detected between plant maturity and partial resistance to the pathogen, with late maturing clones being generally more resistant. Maturity-adjusted resistance, that takes into consideration both plant maturity and resistance, was calculated from residuals of the linear regression between the two traits. Even after adjusting for maturity, the difference in the resistance of clones was still highly significant, indicating that a substantial part of resistance cannot be explained by the effect of maturity. The highest maturity-adjusted resistance was found in the cv. Navajo, while the most susceptible clone was the cv. Pungo. We hope that the present abundance of data about the resistance and maturity of 283 clones will help potato breeders to develop cultivars with improved resistance to V. dahliae.     

Characterization of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> Isolates from Jersey,Channel Islands

Potato production on the island of Jersey, in the English Channel, is dominated by Jersey Royal, a selection of the early cultivar International Kidney. Jersey Royal is very susceptible to Phytophthora infestans, the cause of potato late blight, and Jersey’s climate is frequently conducive to infection. During 2004–2006, isolates of P. infestans were obtained from Jersey Royal plant samples from 41 different sites (24 crops, one infected tuber, 16 volunteers) and from five outdoor tomato crops and characterised by mating type, mtDNA haplotype, Gpi and Pep allozyme genotype, metalaxyl resistance and RG57 fingerprint. A subset of 17 isolates was additionally characterised by simple sequence repeat (SSR). All but one isolate from potato belonged to a single A1 multilocus genotype or a variant. The five isolates from tomato represented three distinct genotypes, none of which was found on potato and included one which was A2 mating type. The populations of P. infestans on potato and tomato in Jersey appear distinct, with that on potato being highly clonal.     

Resistance to <Emphasis Type="Italic">Meloidogyne chitwoodi</Emphasis> Identified in Wild Potato Species

Meloidogyne chitwoodi (Columbia root-knot nematode, CRKN) can cause serious damage in potato production systems, decreasing tuber value in the fresh market and processing industries. Genetic resistance to CRKN was first identified from the wild diploid potato species Solanum bulbocastanum accession SB22 and was successfully introgressed into tetraploid potato breeding material. To expand the base of genetic resistance, 40 plant accessions representing nine wild potato species were screened for their resistance to M. chitwoodi. Greenhouse screening identified fifteen clones from S. hougasii, one clone from S. bulbocastanum, and one clone from S. stenophyllidium with moderate to high levels of resistance against three isolates of M. chitwoodi. Geographical mapping showed that the resistance sources identified in this and previous studies primarily originated in the states of Jalisco and Michoacán in west-central Mexico. These new sources of resistance will be introgressed into elite potato populations to facilitate the development of potato cultivars with durable resistance to M. chitwoodi.     

An <Emphasis Type="Italic">In Vitro</Emphasis> Assay Method for Resistance to Bacterial Wilt (<Emphasis Type="Italic">Ralstonia solanacearum</Emphasis>) in Potato

To develop an in vitro assay method for bacterial wilt resistance in potato, resistant and susceptible standard genotypes were grown in vitro, and different conditions of inoculation with Ralstonia solanacearum phylotype I/biovar 4 were examined. The optimal condition was the inoculation of 6–8 leaf stage plants with a bacterial concentration of 10² CFU ml^?1 and an incubation temperature of 28 °C. Evaluation of stem wilting was more reliable than that of leaf wilting. Using this method, nine genotypes with different resistance levels in the field were evaluated. Lower disease indices were obtained for genotypes with high resistance levels in the field, suggesting that this assay is useful for evaluating bacterial wilt resistance in a controlled environment.     

Impact of Different US Genotypes of <Emphasis Type="Italic">Phytophthora infestans</Emphasis> on Potato Seed Tuber Rot and Plant Emergence in a Range of Cultivars and Advanced Breeding Lines

Seed pieces of different potato cultivars and advanced breeding lines (ABLs) from north central US breeding programmes were inoculated with different genotypes of Phytophthora infestans (US-1, US-1.7, US-8, US-11 and US-14). The effect of these genotypes of P. infestans on seed piece rot severity after re-storage was assessed using an image analysis technique. P. infestans genotypes demonstrated variable ability to cause seed piece rot and to reduce plant emergence measured as final plant stand (%) and the relative area under the plant emergence curve (RAUEPC). The US-8 genotype of P. infestans was the most aggressive genotype, as indicated by tuber rot severity across all cultivars/ABLs tested, followed by US-14 in both years. The US-1, US-1.7 and US-11 genotypes were the least aggressive, causing only moderate seed piece rotting across cultivars/ABLs tested. Similar trends were observed in two field experiments, where the US-8 and US-14 genotypes delayed or reduced emergence. Values of final plant stand (%) and RAUEPC demonstrated that the cultivars/ABLs Atlantic, MSJ453-4Y and Torridon were the least susceptible across all P. infestans genotypes. In both experiments cv. Pike was the most susceptible. Other cultivars/ABLs demonstrated variable responses to different genotypes of P. infestans. No symptoms of P. infestans were observed on emerged plants up to 60 days after planting. The variability of susceptibility of tubers to different genotypes of P. infestans has implications for plant breeding efforts in that the major emphasis in the past has been to breed for foliar resistance, with limited emphasis on the reaction of the tuber. Results from this study suggest that highly aggressive genotypes of P. infestans such as US-8 may lead to severe tuber rotting and deterioration of tubers before emergence, but despite this observation the US-8 genotype is still predominant in North America.     

Performance of Dimethomorph + Mancozeb Applied to Seed Potatoes in Early Management of Late Blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>)

Treating seed potatoes with contact fungicides prevents infection of Phytophthora infestans from tuber to tuber during handling but does not prevent field infections in the young plant. Dimethomorph (DMM) 9% + mancozeb 60% (Acrobat MZ, BASF) can cover both aspects due to the contact and systemic action of mancozeb and DMM, respectively. Experiments were performed under greenhouse and field conditions (three seasons and two locations) with cvs Kennebec, Shepody, Spunta, Russet Burbank, and Ranger Russet. Immediately after cutting, seed pieces were treated with Acrobat MZ at doses equivalent to 2, 3, and 4 kg ha⁻¹. At any dose, Acrobat MZ protected healthy seed tubers against P. infestans infections. Acrobat MZ (4 kg ha⁻¹) also protected the foliage of all cultivars against artificial inoculation of P. infestans, assessed by the detached-leaf method. These effects were observed up to 30 days after emergence in greenhouse experiments and up to 28 days after crop emergence under field conditions in two growing seasons. In cv Spunta, significant foliage protection was observed up to 35 days after field emergence whereas in seed tubers of cv Kennebec planted in sandy soils, significant foliage protection persisted longer. Additional evidence for acropetal translocation of Acrobat MZ in field crops was also found. With this strategy, there is no need to apply contact or systemic fungicides early in the season, and spraying costs and environmental risks are also reduced.     

Impact of Agronomic Strategies (Seed Tuber Pre-sprouting,Cultivar Choice) to Control Late Blight (<Emphasis Type="Italic">Phytophthora infestans</Emphasis>) on Tuber Growth and Yield in Organic Potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis> L.) Crops

In order to optimize potato crop management in organic farming systems, knowledge of crop growth processes in relation to limitations and reductions by nitrogen (N) and disease is necessary. This paper examines the effect of different prevention measures (seed tuber pre-sprouting, choice of cultivars: resistance to Phytophthora infestans; earliness of tuber initiation) against disease-related constraints on yields, depending on the N-mediated growing conditions of organic potato crops. Under conditions of a relatively high N supply, accelerating the early development and tuber initiation of potato crops by seed tuber pre-sprouting (yield increase of c. 18–23%) or the selective choice of cultivars with an earlier tuber initiation (yield increase of c. 0–21%) are the most effective strategies in combating late blight. They bring forward the crop development c. 7–10 days, escaping the negative impact of the fungus on tuber growth. Under conditions of a relatively low N supply, these strategies do not affect final tuber yield. The reason for the differences of the effect depending on N supply of the crops is, that the lower the N supply, the shorter the period of time over which tuber growth takes place, independent of whether P. infestans becomes severe or not. Tuber growth of organic crops low in N has mostly ceased by the time late blight becomes potentially important for limiting yield, with the consequence that preventative measures against P. infestans are meaningless. Surprisingly, the level of cultivar haulm resistance to late blight did not affect tuber yields in years with an early and strong late blight development. Probably, the positive effect of a longer resistance to the fungus (c. 1 week) was counterbalanced by a later tuber initiation (also c. 1 week). In years with a moderate late blight incidence, less susceptible cultivars were able to produce higher tuber yields on fields with a lower N availability (c. 17–20%), probably due to higher N use efficiency and a better match of N mineralization and N uptake. Under conditions of a relatively high N supply, the tested cultivars with a higher haulm resistance to late blight were not able to improve tuber yields.     

Tuber Resistance and Slow-Rotting Characteristics of Potato Clones Associated with the Solanaceae Coordinated Agricultural Project to the US-24 Clonal Lineage of <Emphasis Type="Italic">Phytophthora infestans</Emphasis>

Late blight, caused by Phytophthora infestans, is a devastating disease on potato worldwide and new lineages of the pathogen continue to develop in the U.S. Breeding for resistance is important for economic and environmental purposes. The Solanaceae Coordinated Agricultural Project (SolCAP) focuses on linking allelic variation in genes to valuable traits in elite cultivated potato germplasm. This research assessed the SolCAP diversity panel (206 clones in Washington and 213 clones in Wisconsin) for tuber resistance to the US-24 clonal lineage of P. infestans after potatoes were harvested from fields in Washington and Wisconsin in 2011. This is the first time this germplasm has been evaluated for tuber resistance to P. infestans using a non-intrusive zoospore inoculation technique. Clones with a percent incidence of 30% or less were considered resistant and only eight clones (Palisade Russet, AWN86514–2, MSL268-D, MSM171-A, MSM182–1, MSN230-1RY, Patagonia and Yukon Gem) were characterized as resistant at both locations. These clones have previously demonstrated high to moderate partial foliar resistance to isolates of P.infestans and therefore represent germplasm with both foliar and tuber resistance. Nine clones (AWN86514–2, F66041, MN 18747, MSM 182–1, MSN230-1RY, Modoc, Ama-Rosa, Patagonia and Purple Majesty), were characterized as slow-rotting at both locations with a mean percent internal rot of 75% or less after 33 days of storage. Two clones, MN 18747 and Modoc, are considered to have the highest risk of being a carrier for P. infestans of all the clones evaluated in the SolCAP collection. Not a single clone demonstrated complete tuber resistance to the US-24 strain at both locations.     

Almond (<Emphasis Type="Italic">Prunus dulcis</Emphasis> L.) Protein Quality

Three marketing varieties of almonds; Carmel, Mission, and Nonpareil; were analyzed for proximate composition and protein nutritive quality. Moisture, lipids, protein, ash, sugars, and tannins ranges were 3.05-4.33%, 43.37-47.50%, 20.68-23.30%, 3.74-4.56%, 5.35-7.45%, and 0.12-0.18%, respectively. No detectable hemagglutinating and trypsin inhibitory activities were present in Carmel, Mission, and Nonpareil almonds. Amino acid analyses indicated the sulfur amino acids (methionine + cysteine), lysine, and threonine to be the first, second, and third limiting amino acids in almonds when compared to the recommended amino acid pattern for children 2-5-year old. However, compared to the recommended amino acid pattern for adults, sulfur amino acids were the only limiting amino acids in almonds tested. True Protein Digestibility (% TPD) values for Carmel, Mission, and Nonpareil were 88.55 +/- 1.26, 92.25 +/- 1.05, and 82.62 +/- 1.47, respectively. Protein Digestibility Corrected Amino Acid Scoring (PDCAAS) values suggested almond proteins to be of poor nutritional quality.