Stored Tubers Research Articles

Disease Risk Assessment in Potato Cultures

The aim of our study was to elaborate a system of risk analyse against Phytophtora infestans  (Mont) de Bary attack degree in potato, in connection with climatic factors, in Transylvanian Plane. The risk assessment for potato late blight attack was conducted in tree experimental points located in the counties of Transylvanian Plane: Alba, Cluj, and Mureș. The data were statistically processed using STATISTICA v. 7.0 programme. The analyse of phytosanitary risk assessment consisted in three stages: initiation of the risk assessment, evaluation of the risk analyse, and risk management. Initiation of the risk assessment and evaluation of the ris analyses were performed. The risk management against Phytophtora infestans (Mont) de Bary attack degree in potato initiated involves treatment strategies, using environmentally friendly products, combined to culture appropriate works and strategies, as culture rotations, disinfestations of equipments, and appropriate storage of tubers used for seeding.

Antioxidant activities of the synthesized thiol-contained peptides derived from computer-aided pepsin hydrolysis of yam tuber storage protein, dioscorin

Our previous report showed that yam dioscorin and its peptic hydrolysates exhibit radical scavenging activities; however, the functions of these peptic hydrolases are still under investigation. In this study, the thiol-containing peptides derived from computer-aided simulation of pepsin hydrolysis of dioscorin, namely, KTCGNGME (diotide1), PPCSE (diotide2), CDDRVIRTPLT (diotide3), KTCGY (diotide4), and PPCTE (diotide5) were synthesized to compare their antioxidant activities with GSH and/or carnosine by examining hydroxyl radical scavenging activity by electron spin resonance spectrometry, anti-low-density lipoprotein peroxidation, anti-AAPH-induced hemolysis, and oxygen radical absorbance capacity activity. We found that while all the synthesized diotides showed antioxidant activity, diotide4 exhibited the highest levels. Moreover, all diotides (100μM) showed protective effects against methylglyoxal-induced human umbilical vein endothelial cell death. These results suggest that thiol-containing diotides derived from dioscorin hydrolysis exhibit antioxidant activities and reveal the benefits of yam tuber as an antioxidant-rich food.

A mass spectrometric method for quantifying C3 and C6 phosphorylation of starch

The glucosyl residues comprising starch can be phosphorylated at either the C3 or the C6 position of the molecule because of the activities of two distinct dikinase enzymes. After hydrolysis of the starch, the C6 phosphorylation is easy to measure using a routine enzyme assay for glucose 6-phosphate, but the C3 phosphorylation is more difficult to assay. A mass spectrometric (MS) method has been developed that, in a single run, can distinguish and quantify the glucose 3-phosphate and glucose 6-phosphate produced by hydrolysis of starch and can also measure the glucose content to give an accurate estimate of the starting material. The MS method involves quantification by LC/MS with external standards, using normal-phase hydrophilic interaction liquid chromatography and selective reaction monitoring. The MS method has been used to determine degrees of starch phosphorylation in a diverse group of potato lines, revealing threefold differences in phosphorylation between high- and low-phosphate lines. The method was also used to show that cold storage of potato tubers for up to 24weeks had little substantive effect on the levels of starch phosphorylation. MS provided an effective and efficient means of determining both the C6 and the C3 phosphorylation of starch.

Concentrations of some macroelements in potato tubers stored at 4ºC and 8ºC

The objective of the investigations was to evaluate changes in concentrations of total protein (nitrogen x 6.25), phosphorus and potassium in tubers of two potato cultivars: Rywal and Saturna, as dependent on the application of herbicides (Afalon 50 WP, Azogard 50 WP, Sencor 70 WG, and Apyros 75 WG), after harvest and after 3 and 6 months of storage. Concentrations of total protein, phosphorus and potassium in potato tubers were strongly conditioned by genetic features of the cultivars. The tubers collected from the objects sprayed with the herbicides over the vegetation period contained by 3.7%, 8.1%, and by 3.5% more protein, phosphorus and potassium, respectively, than those nursed exclusively mechanically. The corresponding values in the latter case reached respectively (means for the objects with the use of herbicides) 24.4 g kg-1, 660.5 mg kg-1 and 5351.3 mg kg-1. After 3 and 6 months of storage at 8°C, the content of nitrogen was significantly lower. Similarly, tubers stored at 4oC contained significantly less nitrogen, but not earlier than after 6 months. In contrast, concentrations of phosphorus and potassium did not change significantly over the storage of tubers in chambers with the lower temperature (4°C).

First Report of Phytophthora cryptogea on Potato Tubers in Turkey.

Potato (Solanum tuberosum L.) is an important agricultural crop in Turkey, with 150,000 ha and a production of about 4 million tons per year. There are numerous pathogens that limit potato production. In 2011, a new disease was observed on the potato cultivar Ramos in Erzincan Province in Turkey. Disease symptoms were similar to pink rot. Infected tubers had a rubbery texture and when cut, infected areas were cream colored. When the infected area was exposed to air for 10 min, the color turned to salmon pink and after 15 to 20 min it turned black. A funguslike organism was isolated from diseased tubers on carrot agar. The colonies were uniform and there were no petaloid patterns. Soil extract solution was prepared using 1,000 ml of distilled water and 15 g of field soil according to Jeffers and Aldwinckle (2) and used for sporangia production. Agar discs from 5-day-old colonies were placed in soil extract solution and incubated under continuous fluorescent light at room temperature. Sporangia began forming after 24 h and were abundant after 48 h. Oogonia were not observed on corn meal agar plates supplemented by beta-cytosterol. Sporangia were nonpapillate and noncaducous, mostly ovoid, but some were obpyriform. The mean size of the sporangia was 43 ± 4 × 30 ± 2 μm. The Oomycete was identified as Phytophthora cryptogea Pethybr. & Laff., according to its morphological characters (1). Isolate identity was confirmed by sequence analysis of the ribosomal DNA internal transcribed spacer using primers ITS1 and ITS4. The ITS sequence was a 99% match to P. cryptogea strains (GenBank Accession Nos. AF087475.1, AF228101.1, GU111626.1, AY995400.1, GU111624.1, AY995400.1, and GU111624.1). The isolate from Turkey was deposited in GenBank as Accession No. JQ071495. Pathogenicity tests were conducted on potato tubers (cv. Ramos). Tubers were surface disinfected with 0.5% NaOCl for 5 min., then washed twice with sterile water. When dried, 20 tubers were wounded with a shallow cut made parallel to the surface by a sterile knife. An agar disc from a 1-week-old colony was placed on the artificial wound on each of 10 potatoes. For controls, agar discs without fungal mycelia were placed on the wounds of the other 10 potatoes. Each inoculated and uninoculated potato was incubated for 5 days at 24°C in the dark and in separate plastic cups containing moistened filter paper at the bottom. Tubers were then cut open and examined for disease. Cream colored lesions were first observed; after 10 min lesions turned salmon pink and after 20 min they turned black. Cream colored lesions were not observed inside uninoculated potatoes. To the best of our knowledge, this is the first report of P. cryptogea causing pink rot of potato in Turkey. Pink rot disease of potato is commonly caused by P. erytroseptica, but P. cryptogea can cause similar symptoms (3) and severe tuber rot. This pathogen could cause important losses of potato in Turkey, especially during storage of tubers.

Molecular and physiological properties associated with zebra complex disease in potatoes and its relation with Candidatus Liberibacter contents in psyllid vectors.

Zebra complex (ZC) disease on potatoes is associated with Candidatus Liberibacter solanacearum (CLs), an α-proteobacterium that resides in the plant phloem and is transmitted by the potato psyllid Bactericera cockerelli (Šulc). The name ZC originates from the brown striping in fried chips of infected tubers, but the whole plants also exhibit a variety of morphological features and symptoms for which the physiological or molecular basis are not understood. We determined that compared to healthy plants, stems of ZC-plants accumulate starch and more than three-fold total protein, including gene expression regulatory factors (e.g. cyclophilin) and tuber storage proteins (e.g., patatins), indicating that ZC-affected stems are reprogrammed to exhibit tuber-like physiological properties. Furthermore, the total phenolic content in ZC potato stems was elevated two-fold, and amounts of polyphenol oxidase enzyme were also high, both serving to explain the ZC-hallmark rapid brown discoloration of air-exposed damaged tissue. Newly developed quantitative and/or conventional PCR demonstrated that the percentage of psyllids in laboratory colonies containing detectable levels of CLs and its titer could fluctuate over time with effects on colony prolificacy, but presumed reproduction-associated primary endosymbiont levels remained stable. Potato plants exposed in the laboratory to psyllid populations with relatively low-CLs content survived while exposure of plants to high-CLs psyllids rapidly culminated in a lethal collapse. In conclusion, we identified plant physiological biomarkers associated with the presence of ZC and/or CLs in the vegetative potato plant tissue and determined that the titer of CLs in the psyllid population directly affects the rate of disease development in plants.

Owyhee Russet: A Variety with High Yields of U.S. No. 1 Tubers, Excellent Processing Quality, and Moderate Resistance to Fusarium Dry Rot (Fusarium solani var. coeruleum)

Owyhee Russet (AO96160-3) originated from a cross between A89384-10 and A89512-3 in 1996. Owyhee Russet was released in 2009 by Oregon State University, in cooperation with the USDA-ARS and the Agricultural Experiment Stations of Idaho and Washington and is a product of the Northwest Potato Variety (Tri-State) Development Program. Owyhee Russet has semi-erect medium sized vines with medium to late maturity. The tubers are long, with a tan skin, medium russeting, and attractive tuber appearance for fresh market. Owyhee Russet was evaluated in several locations across the Northwest for more than 15 years. Total yield of Owyhee Russet is similar to that of Russet Burbank and Ranger Russet but significantly higher than Russet Norkotah. U.S. No.1 tuber yield of Owyhee Russet is significantly higher than Russet Burbank and Russet Norkotah, resulting in substantially higher marketable yield. Owyhee Russet tubers have significantly higher specific gravity than Russet Burbank and Russet Norkotah. Fry color following tuber storage at 4°C and 9°C is significantly lighter for Owyhee Russet than the comparison varieties. Relative strengths include high yield with a very high proportion of U.S. No.1 tubers, good tuber appearance and excellent processing quality, resistance to cold sweetening, common scab and Fusarium dry rot. Weaknesses include susceptibility to foliar and tuber late blight and susceptibility to metribuzin herbicide injury. Allelic patterns of five SSR markers have shown that Owyhee Russet has a distinctive DNA genetic fingerprint from its russet type reference varieties which are Ranger Russet, Russet Burbank, and Russet Norkotah.

Processing quality of potato tubers produced during autumn and spring and stored at different temperatures

The objective of this work was to access processing quality of potato clones (Solanum tuberosum) Asterix, SMINIA793101-3, and Missaukee cultivated during spring and autumn growing seasons and stored at 4, 8, 12, or 25ºC. Clones grown in spring had shorter dormancy than clones grown in autumn. Potato tubers grown in spring and autumn and stored at 4ºC, as well as tubers grown in autumn and stored at 8ºC had no sprouting for six months. Among clones grown in autumn, the longest dormancy period during storage at 12ºC was observed in the clone SMINIA793101-3, and at 25ºC was in the clone Asterix. In potato tubers grown in spring, the longest dormancy period during storage at 8ºC was observed in the clones SMINIA793101-3 and Missaukee, at 12ºC was in the clone SMINIA793101-3, and at 25ºC was in the clones Asterix and Missaukee. Potato tubers grown in spring had higher dry matter, starch content, and respiration rates, and lower reducing sugar, and total polyphenol content. The only exception was the Missaukee that had similar dry matter content in both growing seasons. In general, storage at 4ºC and 8ºC were more effective in reducing respiration rates and maintaining higher dry mater content of potato tubers. Chip darkening, reducing sugars, and total polyphenol content increased during storage of potato tubers. Darker chip color, higher reducing sugars and polyphenol content were obtained during storage at temperatures of 4ºC and 8ºC. These results suggest that the best storage temperature depends on genotype, tuber physiological age, and growing conditions.

Crystallization and preliminary X-ray crystallographic analysis of dioscorin from Dioscorea japonica.

Dioscorin, the major tuber storage protein in yam, has been reported to possess carbonic anhydrase, trypsin inhibitor, angiotensin-converting enzyme (ACE) inhibitor, free-radical scavenger, dehydroascorbate reductase and monodehydroascorbate reductase activities. Recent research has also found that dioscorin can enhance immune modulation via the toll-like receptor 4 (TLR-4) signal transduction pathway in RAW 264.7 cells, murine bone-marrow cells and human monocytes ex vivo. Resolving the structure of dioscorin would help in better understanding its activities and would provide clues to understanding the mechanism of its multiple functions. The full-length protein (residues 1-246) with an additional His(6) tag at the N-terminus was expressed in Escherichia coli Rosetta (DE3) cells. After His-tag cleavage and purification, the protein was crystallized by the sitting-drop vapour-diffusion method at 278 K. An X-ray diffraction data set was collected to a resolution of 2.11 Å using a synchrotron X-ray source. The crystal belonged to space group C222(1), with unit-cell parameters a = 83.5, b = 156.8, c = 83.6 Å, and was estimated to contain two protein molecules per asymmetric unit.

Biological Activities and Applications of Dioscorins, the Major Tuber Storage Proteins of Yam

Yam tubers, a common tuber crop and an important traditional Chinese medicine in Taiwan, have many bioactive substances, including phenolic compounds, mucilage polysaccharides, steroidal saponins and proteins. Among the total soluble proteins, 80% of them are dioscorins. In the past two decades, many studies showed that dioscorins exhibited biological activities both in vitro and in vivo, including the enzymatic, antioxidant, antihypertensive, immunomodulatory, lectin activities and the protecting role on airway epithelial cells against allergens in vitro. Some of these activities are survived after chemical, heating process or enzymatic digestion. Despite of lacking the intact structural information and the detail action mechanisms in the cells, yam dioscorins are potential resources for developing as functional foods and interesting targets for food protein researchers.

Adaptation to flooding during emergence and seedling growth in rice and weeds, and implications for crop establishment.

Direct seeding of rice is being adopted in rainfed and irrigated lowland ecosystems because it reduces labour costs in addition to other benefits. However, early flooding due to uneven fields or rainfall slows down seed germination and hinders crop establishment. Conversely, early flooding helps suppress weeds and reduces the costs of manual weeding and/or dependence on herbicides; however, numerous weed species are adapted to lowlands and present challenges for the use of flooding to control weeds. Advancing knowledge on the mechanisms of tolerance of flooding during germination and early growth in rice and weeds could facilitate the development of improved rice varieties and effective weed management practices for direct-seeded rice. Rice genotypes with a greater ability to germinate and establish in flooded soils were identified, providing opportunities to develop varieties suitable for direct seeding in flooded soils. Tolerance of flooding in these genotypes was mostly attributed to traits associated with better ability to mobilize stored carbohydrates and anaerobic metabolism. Limited studies were undertaken in weeds associated with lowland rice systems. Remaining studies compared rice and weeds and related weed species such as Echinochloa crus-galli and E. colona or compared ecotypes of the same species of Cyperus rotundus adapted to either aerobic or flooded soils. Tolerant weeds and rice genotypes mostly developed similar adaptive traits that allow them to establish in flooded fields, including the ability to germinate and elongate faster under hypoxia, mobilize stored starch reserves and generate energy through fermentation pathways. Remarkably, some weeds developed additional traits such as larger storage tubers that enlarge further in deeper flooded soils (C. rotundus). Unravelling the mechanisms involved in adaptation to flooding will help design management options that will allow tolerant rice genotypes to adequately establish in flooded soils while simultaneously suppressing weeds.

EFFECTS OF STORAGE ON THE CHEMICAL COMPOSITION AND FOOD QUALITY OF YAM

ABSTRACT The effects of storage (4 months) on the chemical composition of yam tubers and its relation to textural quality of a major yam food product (pounded yam) were studied using six varieties of Dioscorea rotundata and D. alata. Chemical composition of both freshly harvested tubers and those stored for 4 months were determined by standard methods of analysis. Sugar, nonstarchy carbohydrate and dry matter contents of the tubers increased during storage while the starch, fat and protein contents decreased. Pounded yam made from stored yam tubers were of better textural quality than those from freshly harvested ones. Storage of yam tubers could be an effective way of improving the textural quality of pounded yam. This is important for processing yam tubers into value-added products. PRACTICAL APPLICATION The results of this research provide a scientific basis for differences in the food quality of food products made from freshly harvested and stored yam tubers. Storage is important to the industrial processing of value-added products (e.g., instant yam flour, yam flakes) from yam tubers. This study will help processors in selecting appropriate raw materials that will give optimal food quality that is expected and will be accepted by consumers.

The sprout inhibitor 1,4-dimethylnaphthalene induces the expression of the cell cycle inhibitors KRP1 and KRP2 in potatoes

The suppression of sprout growth is critical for the long-term storage of potato tubers. 1,4-Dimethylenapthlene (DMN) is a new class of sprout control agent but the metabolic mode of action for this compound has yet to be elucidated. Changes in transcriptional profiles of meristems isolated from potato tubers treated with the DMN were investigated using an Agilent 44K 60-mer-oligo microarray. RNA was isolated from nondormant Russet Burbank meristems isolated from tubers treated with DMN for 3days or activated charcoal as a control. RNA was used to develop probes that were hybridized against a microarray developed by the Potato Oligo Chip Initiative. Analysis of the array data was conducted in two stages: total array data was examined using a linear model and the software Limma and pathway analysis was conducted by linking the potato sequences to the Arabidopsis thaliana. DMN elicited a change in a number of transcripts associated with cold responses, water regulation, salt stress, and osmotic adjustment. DMN also resulted in a repression of cyclin or cyclin-like transcripts. DMN also resulted in a 50% decrease in thymidine incorporation suggesting a repression of the S phase of the cell cycle. Quantitative real-time polymerase chain reaction analysis demonstrated that DMN increased transcripts for the cell cycle inhibitors KRP1 and KRP2. We conclude the DMN results in alteration of genes associated with the maintenance of a G1/S phase block possibly through the induction of the cell cycle inhibitors KRP1 and KRP2.

Cloning of genes and enzymatic characterizations of novel dioscorin isoforms from Dioscorea japonica

Dioscorin, the major tuber storage protein of yam, has been shown to possess carbonic anhydrase, trypsin inhibitor, dehydroascorbate reductase, and monodehydroascorbate reductase activities. In the present study, dioscorin from Dioscorea japonica was confirmed as a glycoprotein using the enhanced concanavalin A-peroxidase staining method, and the protein was shown to have both N- and O-glycans. Following the gene cloning, four full-length isoforms of dioscorin were expressed in Escherichia coli and purified by affinity purification and anion-exchange chromatography for structural and biochemical experiments. It was clearly observed that the recombinant dioscorins had carbonic anhydrase, trypsin inhibitor, dehydroascorbate reductase, and monodehydroascorbate reductase activities. However, the dehydroascorbate reductase and monodehydroascorbate reductase activities were markedly decreased in recombinant dioscorins compared with native dioscorin. The decreased activities were closely related to the loss of the glycosylation from the protein.

Extensive post‐translational processing of potato tuber storage proteins and vacuolar targeting

Potato tuber storage proteins were obtained from vacuoles isolated from field-grown starch potato tubers cv. Kuras. Vacuole sap proteins fractionated by gel filtration were studied by mass spectrometric analyses of trypsin and chymotrypsin digestions. The tuber vacuole appears to be a typical protein storage vacuole absent of proteolytic and glycolytic enzymes. The major soluble storage proteins included 28 Kunitz protease inhibitors, nine protease inhibitors 1, eight protease inhibitors 2, two carboxypeptidase inhibitors, eight patatins and five lipoxygenases (lox), which all showed cultivar-specific sequence variations. These proteins, except for lox, have typical endoplasmic reticulum (ER) signal peptides and putative vacuolar sorting determinants of either the sequence or structure specific type or the C-terminal type, or both. Unexpectedly, sap protein variants imported via the ER showed multiple molecular forms because of extensive and unspecific proteolytic cleavage of exposed N- and C-terminal propeptides and surface loops, in spite of the abundance of protease inhibitors. Some propeptides are potential novel vacuolar targeting peptides. In the insoluble vacuole fraction two variants of phytepsin (aspartate protease) were identified. These are most probably the processing enzymes of potato tuber vacuolar proteins. Database Proteome data have been submitted to the PRIDE database under accession number 17707.

Losses of Ascorbic Acid During Storage of Fresh Tubers, Frying, Packaging and Storage of Potato Crisps from Four Kenyan Potato Cultivars

Losses of Ascorbic Acid During Storage of Fresh Tubers, Frying, Packaging and Storage of Potato Crisps from Four Kenyan Potato Cultivars

Rheological and structural properties of starches from γ-irradiated and stored potatoes

Starch was extracted from irradiated and stored potato tubers and the properties were compared to CIPC (chlorpropham) treated tubers. The granule properties and dynamic viscoelasticity in temperature ramp and frequency sweep modes were studied while heating the samples. Starch structural characteristics were investigated by high performance anion exchange chromatography (HPAEC) and Fourier transform infrared spectroscopy (FTIR). Gamma-irradiation of potato tubers at a dosage of 0.1kGy induced some degradation of starch molecules, resulting in earlier swelling of starch granules, and greater extents of amylose and total carbohydrate leaching. The early swelling phenomenon was also enhanced with tuber storage time. The retrogradation rate and extent for a concentrated starch gel also increased with tuber storage time whereas γ-irradiation delayed the gel retrogradation. Sprout inhibiting methods could be selected based on the specific processing and texture requirements of the end products.

Potato soil-borne diseases. A review

Potato crop is the fourth main food crop in the world and it will certainly feed a big part of the global population in the next years. The economical outlets for this crop are great; however, numerous diseases either soil- or air-borne can cause huge losses in the production. Worldwide, about 40 soil-borne diseases affect potato and cause severe damages especially on tubers, the economically most important part of the plant. The occurrence and development of soil-borne diseases depend on very diverse factors affecting either the pathogen or the plant. Favorable conditions for potato diseases development are frequently the same as the conditions needed for potato growth: temperature between 10°C and 25°C, high humidity, medium pH, etc. Adapted cultural practices such as a rotation longer than 4 years, appropriate fertilization and water management, an adapted delay between haulm killing and harvest, and dry and cool conditions for tuber storage are good ways to control potato diseases. In most cases, potato pathogens develop specific survival forms, dissemination ways and host penetration methods. The genetic variability of the pathogens implies the use of adapted diagnostic and control methods. Decision support systems developed to predict yield losses allow choosing good control methods such as the use of healthy seeds, adapted pesticides, cultural practices, and biological control agents for each potato disease. The complexity of the interactions between a pathogen and its host, influenced by biotic and abiotic factors of the environment, make the control of the diseases often very difficult. However, deep knowledge of pathosystems allows setting up integrated pest management systems allowing the production of healthy and good quality potatoes.

Systematic analysis of potato acid invertase genes reveals that a cold-responsive member, StvacINV1, regulates cold-induced sweetening of tubers

Acid invertase is believed to play a regulatory role during plant developmental processes and to respond to environmental stimuli. The expression profiles of the entire acid invertase family are not yet available for potato. By searching existing databases, it was determined that there are at least six acid invertase genes in potato, including four cell-wall invertase genes and two vacuolar invertase genes. They were subjected to comparative expression profiling in various organs of potato plants and in stored tubers to exploit their potential functions. The results revealed that each gene exhibited a unique expression pattern, which differed in transcript abundance or showed organ-specific features, pointing to the possible involvement of individual genes in plant development. The vacuolar invertase gene StvacINV1 had the highest expression level among three genes detected in the potato tubers. Further storage experiments showed that StvacINV1 was strongly induced by low temperatures, which is consistent with glucose accumulation in cold-stored tubers. Suppression of StvacINV1 by the antisense transformation in potato confirmed that lower StvacINV1 transcript abundance in transgenic tubers is related to lower reducing sugar content and lighter chip color in comparison with the wild type. The evidence strongly suggests that StvacINV1 is a gene involved in regulation of cold-induced sweetening of potato tubers. This provides an avenue for studying the mechanism involved in the regulation of the cold-induced sweetening trait and for agronomic enhancement.

Long-Term Cold Storage Suppress the Development of Tuber Necrosis Caused by PVYNTN

The aim of this work was to determine the influence of different storage temperatures and tuber maturity on development of tuber necrosis caused by Potato virus Y NTN . Infected tubers of the cultivar Igor of two different maturity groups and non-infected tubers were randomly distributed into 12 storage regimes. Control treatments were stored at constant temperatures of 4, 13 and 24°C. Tubers in the other nine treatments were stored at 4°C at the beginning of experiment and transferred to 24°C after 1 to 29 weeks after harvest. High storage temperatures enhanced the development of tuber necrosis. Prolongation of low temperature storage delayed the development of necrosis and reduced the number of tubers with necrosis. When tubers were stored at low temperature for 21 weeks or more, no necrosis developed after transfer to high temperature. We could not confirm the effect of maturity on development of necrosis caused by PVYNTN.