First Detection of Potato Spindle Tuber Viroid in Natural Isolates of Potato Blight Agent Phytophthora infestans.

Phytophthora infestans is the oomycete that causes potato blight, an important disease. The potato spindle tuber viroid (PSTVd) is a dangerous pathogen of many plants, including potato. We have previously shown that PSTVd can be transmitted from infected potato plants into the Ph. infestans mycelium, replicated within the mycelium, and then transmitted to other potato plants upon their infection with Ph. infestans in laboratory conditions. The objective of this work was to check the hypothesis that PSTVd transmission, preservation, and replication in Ph. infestans are possible to occur in natural conditions during long-term coevolution of the host and pathogen in the Solanum spp.-Ph. infestans system. A screening test for PSTVd was performed in 111 natural Ph. infestans isolates obtained from potato plants, which represented various cultivars, had signs of potato blight, and were collected from industrial potato fields of the Moscow, Vologda, and Bryansk regions and breeding and variety test plots of the St. Petersburg and Moscow regions in 2020 and 2022. Using RT-PCR with PSTVd-specific primers, 42 Ph. infestans isolates collected in 2020 were tested after five passages and 69 Ph. infestans isolates collected in 2022, after a single passage on rye agar. Diagnostic amplicons were detected in 8 and 50 isolates, respectively. Some of the amplicons were visually assessed as minor amplification products, apparently resulting from nonspecific priming on a host Ph. infestans gene, which codes for a hypothetical protein-coding mRNA in Ph. infestans and other oomycetes. Eight amplicons were sequenced to verify the PSTVd presence in Ph. infestans isolates. Three amplicons corresponded to the complete PSTVd genome and five, to its part (~260 bp). The nucleotide sequences of cloned amplification products were identified to species in the BLAST system and deposited in GenBank. The amplicons obtained with the PSTVd-specific primers were identified as PSTVd sequences in all Ph. infestans isolates examined. The majority of the nucleotide sequences were phylogenetically related to BLAST sequences of PSTVd strains originating from Russia; several strains showed similarity to strains from other countries (France, China, and West African countries). The results demonstrate that PSTVd was for the first time detected in natural (field) Ph. infestans isolates and offer new opportunities for studying the intricate multilevel host-parasite interactions.

Transmission of potato spindle tuber viroid between Phytophthora infestans and host plants.

Potato spindle tuber viroid (PSTVd) is a naked, circular, single-stranded RNA (356-363 nucleotides in length) which lacks any protein-coding sequences. It is an economically important pathogen and is classified as a high-risk plant quarantine disease. Moreover, it is known that PSTVd is mechanically transmitted by vegetative plant propagation through infected pollen, and by aphids. The aim of this study is to determine the possibility of viroid transmission by potato pathogen Phytophthora infestans (Mont.) de Bary. PSTVd-infected (strain VP87) potato cultivars Gala, Colomba, and Riviera were inoculated with P. infestans isolate PiVZR18, and in 7 days, after the appearance of symptoms, re-isolation of P. infestans on rye agar was conducted. RT-PCR diagnostics of PSTVd in a mixture of mycelia and sporangia were positive after 14 days of cultivation on rye agar. The PSTVd-infected P. infestans isolate PiVZR18v+ was used to inoculate the healthy, viroid-free plants of potato cv. Gala and tomato cv. Zagadka. After 60 days, an amplification fragment of PSTVd was detected in the tissues of one plant of tomato cv. Zagadka by RT-PCR with the primer set P3/P4, indicating successful transmission of PSTVd by P. infestans isolate PiVZR18v+. This result was confirmed by sequencing of the RT-PCR amplicon with primers P3/P4. The partial sequence of this amplicon was identical (99.5 %) to PSTVd strain VP87. RT-PCR showed the possibility of viroid stability in a pure culture of P. infestans isolate PiVZR18v+ after three consecutive passages on rye agar. PSTVd was not detected after the eighth passage on rye agar in P. infestans subculture. These results are initial evidence of potato viroid PSTVd being bidirectionally transferred between P. infestans and host plants.

Hybridization between Pyrenophora teres Forms in Natural Populations of Russia and the Republic of Belarus.

Two forms of the pathogenic fungus Pyrenophora teres, P. teres f. teres (a net form) and P. teres f. maculata (a spot form), cause different disease signs, net or spot blotch, on barley leaves. The net form of P. teres is widespread wherever barley is cultivated, while the spot form was first identified in Krasnodar krai of Russia in 2011 and Brest oblast of Belarus in 2016. The two forms of the pathogen easily mate each other in laboratory conditions, but their hybrids either do not form or are difficult to detect in nature. The question as to whether hybrids between the net and spot forms are produced and maintained in natural populations is pressing and bears applied significance because different genes determine resistance to the different P. teres forms in barley. Hybrid forms may be virulent to resistance donors used in breeding. The objective of this work was to search Russian and Belarussian natural populations for hybrids between P. teres f. teres and P. teres f. maculata with the use of new form-specific markers, Ptt and Ptm. The study included 138 single-conidium isolates from four P. teres f. maculatа and four P. teres f. teres populations. The isolates were collected from commercial barley plantations of Leningrad oblast, Krasnodar krai (Russia), and Brest oblast (Belarus) from 2013 to 2016. A genotyping with 10 form-specific markers was performed in all isolates. Several isolates were found to combine markers of both of the P. teres forms and were conventionally identified as hybrids between the forms. Hypotheses were advanced to explain the occurrence of hybrids in natural populations. The most plausible hypothesis suggests that sexual or somatic hybridization between the two forms coexisting in barley plantations accounts for the origin of the P. teres isolates that combine markers specific to P. teres f. teres and markers specific to P. teres f. maculata in their genomes. It is also possible that a third, possibly ancestral, intermediate form was preserved during divergence in the species P. teres.

Potato spindle tuber viroid.

Viroids belong to a very interesting class of molecules attracting researchers in phytopathology and molecular evolution. Here we review recent literature data concerning the genetics of Potato spindle tuber viroid (PSTVd) and the mechanisms related to its pathological effect on the host plants. PSTVd can be transmitted vertically through microspores and macrospores, but not with pollen from another infected plant. The 359 nucleotidelong genomic RNA of PSTVd is highly structured and its 3D-conformation is responsible for interaction with host cellular factors to mediate replication, transport between tissues during systemic infection and the severity of pathological symptoms. RNA replication is prone to errors and infected plants contain a population of mutated forms of the PSTVd genome. Interestingly, at 7 DAI, only 25 % of the newly synthesized RNAs were identical to the master copy, but this proportion increased to up to 70 % at 14 DAI and remained the same afterwards. PSTVd infection induces the immune response in host plants. There are PSTVd strains with a severe, a moderate or a mild pathological effect. Interestingly, viroid replication itself does not necessarily induce strong morphological or physiological symptoms. In the case of PSTVd, disease symptoms may occur due to RNA-interference, which decreases the expression levels of some important cellular regulatory factors, such as, for example, potato StTCP23 from the gibberellic acid pathway with a role in tuber morphogenesis or tomato FRIGIDA-like protein 3 with an early flowering phenotype. This association between the small segments of viroid genomic RNAs complementary to the untranslated regions of cellular mRNAs and disease symptoms provides a way for new resistant cultivars to be developed by genetic editing. To conclude, viroids provide a unique model to reveal the fundamental features of living systems, which appeared early in evolution and still remain undiscovered.

[New genetic tools for plant defense against parasitic nematodes]. / Новые генетические Ñ‚ÐµÑ Ð½Ð¾Ð»Ð¾Ð³Ð¸Ð¸ защиты растений от Ð¿Ð°Ñ€Ð°Ð·Ð¸Ñ‚Ð¸Ñ‡ÐµÑÐºÐ¸Ñ Ð½ÐµÐ¼Ð°Ñ‚Ð¾Ð´.

. Nematodes belong to economically important pests. Here we reviewed the recent data on molecular mechanisms of plant resistance to cyst and gall nematodes including the most devastating Globodera rostochiensis, G. pallida, Heterodera schachtii, Meloidogyne chitwoodi, and M. incognita. The Golden Potato Cyst Nematode (G. rostochiensis, GPCN) may be taken as an example of an economically important pest: in Russia, it occurs in 61 regions with a total area of 1.8 million ha and may cause the yield loss from 19 to 90 %. The biological characteristics of sedentary nematodes makes their agrotechnical control problematic, i.e. the GPCN cysts remain dormant in soil for many years until a susceptible host appears, whereas nematicides are either toxic or inefficient. Introgression of resistance genes (R-genes) from related cultivated or wild species is likely to be the most appropriate way for their biocontrol. The life cycle of sedentary nematodes is based on juveniles' penetration into the host root where they reprogram plant cells into a syncytium or the so-called 'giant cells' and inhibit the plant defense response. Molecular mechanisms of plant-nematode interaction are unusual and this phenomenon provides a very interesting model for the investigation of plant morphogenesis control as well as for the development of new genetic instruments of biocontrol. Here we reviewed recent publications on plant parasitic nematode effectors used for hijacking of the plant immune system, data on R-genes and molecular mechanisms of their activities. In addition, host-induced gene silencing (HIGS) is discussed as a perspective mechanism for nematode biocontrol. HIGS is based on the RNA interference in the cells of the host plant addressed against the nematode genes important for their development and productivity. Several recent investigations demonstrated efficiency of HIGS against sedentary nematodes.

Occurrence and distribution of viruses infecting potato in Russia.

Potato viral disease has been a major problem in potato production worldwide including Russia. Here, we detected Potato Virus M (PVM), P (PVP), S (PVS), Y (PVY), and X (PVX) and Potato Leaf Roll Virus (PLRV) by RT-PCR on potato leaves and tubers from the Northwestern (NW), Volga (VF), and Far Eastern (FE) federal districts of Russia. Each sample was co-infected with up to five viruses. RT-PCR disclosed all six viruses in NW, three in VF, and five in FE. Phylogenetic analyses of PVM and PVS strains resolved all PVM isolates in Group O (ordinary) and all PVS isolates in Group O. Seven PVY strains were detected, and they included only recombinants. PVY recombinants were thus the dominant potato virus strains in Russia, although they widely varied among the regions. Our research provides insights into the geographical distribution and genetic variability of potato viruses in Russia.

[Resistance to causal agents of late blight and golden potato nematode of the modern cultivars of seed potatoes and their phytosanitary status in various agroclimatic zones of the European part of Russia]. / Устойчивость к возбудителям фитофтороза и глободероза современного сортимента семенного картофеля и его фитосанитарное состояние в Ñ€Ð°Ð·Ð»Ð¸Ñ‡Ð½Ñ‹Ñ Ð°Ð³Ñ€Ð¾ÐºÐ»Ð¸Ð¼Ð°Ñ‚Ð¸Ñ‡ÐµÑÐºÐ¸Ñ Ð·Ð¾Ð½Ð°Ñ ÐµÐ²Ñ€Ð¾Ð¿ÐµÐ¹ÑÐºÐ¾Ð¹ части России.

The active expansion of foreign potato cultivars on the territory of the Russian Federation has led to a change in the dominant pathogen species and to the emergence of new pathotypes of causal agents of harmful potato diseases. The aim of the study was to evaluate resistance to Phytophthora infestans and Globodera rostochiensis of modern potato cultivars and determine the distribution of fungal and oomycetic diseases on potato cultivars in various agroclimatic zones of Russia. The resistance of 41 foreign cultivars was evaluated to pathotype Ro1 G. rostochiensis and to isolate VZR17 P. infestans with virulence genes 1.2.3.4.5.6.7.8.9.10.11. Resistant to G. rostochiensis were 38 cultivars. 57R marker of the H1 gene conferring resistance to the Ro1 pathotype of G. rostochiensis was detected in 96.6 % of the nematode resistant cultivars studied; susceptible varieties did not possess this marker. Absolute resistance to the causative agent of late blight was demonstrated by the cultivars Alouette and Sarpo Mira (score 9); high levels of resistance (score 6 and 7) were determined for the cultivars Evolution, Red Fantasy and Ricarda. The cultivars Baltic Rose, Damaris, Desiree, Gala, Labella, Laperla, Mia, Sanibel, Zekura, Queen Anne, Red Lady and '7 for 7' were classified as susceptible, although the characteristics of originators indicated average resistance to late blight. A phytopathological test was conducted on 92 samples of 39 varieties of seed potatoes from four federal districts of the Russian Federation: Volga, NorthWest, Central and North Caucasus. Rhizoctonia solani, Fusarium spp. and Helminthosporium solani are most common on all varieties. 100 % defeat of tubers by H. solani was recorded in various regions on the cultivars Red Scarlett, Evolution, Labella, Colombo, Gala and Nevsky. Widespread Colletotrichum coccodes on tubers of the elite and 2nd reproductions of the potato cultivar Red Scarlett (50.0-71.4 %) was recorded in the Central District.

[Quarantine nematode species and pathotypes potentially dangerous for domestic potato production: populations diversity and the genetics of potato resistance]. / Потенциально опасные для отечественного картофелеводства карантинные виды и патотипы нематод: изменчивость популяций и генетика устойчивости картофеля.

The review considers quarantine species and nematode pathotypes potentially dangerous for domestic potato production. Potatoes are affected by more than 30 types of parasitic nematodes, but the review focuses on the most harmful representatives of genera that cause great damage to potato production: Globodera, Ditylenchus, Nacobbus and Meloidogyne. Phytopathological and molecular methods of identification of species and pathotypes and the main achievements in studying the population variability of parasitic potato nematodes were analyzed. It was shown that due to the peculiarities of the life cycle of nematodes and lability of their genomes, the genetic variability of these organisms is very high, which creates a threat of forming new pathogenic genotypes of the parasites. The information about the intra- and interpopulation variability of nematodes is important for studying the ways of introduction and distribution of separate species, as well as for searching for the correlations of molecular markers with the pathotype. Phylogenetic studies based on modern data on genetic variability of populations have allowed to reveal species complexes in Globodera pallida (Stone) Behrens and Nacobbus aberrans (Thorne) Thorne & Allen (sensu lato), including cryptic species. The main components of successful protection preventing a wide distribution of parasitic nematodes are quarantine measures, agricultural techniques, biological methods of protection and cultivation of resistant cultivars. Special attention in the review is paid to the breeding of potato cultivars with durable resistance to various nematode pathotypes, because the cultivation of such varieties is the most ecologically safe and economically advantageous way to prevent epiphytoties. Currently, significant progress has been made in the genetic protection of potato cultivars, especially against cyst-forming nematodes. The review provides data on sources of potato resistance to parasitic nematodes identified in collections of wild and cultivated species. Data on identified R-gens and QTL of resistance that have been introduced into breeding varieties using different methods and approaches are analyzed. The literature data on the study of structural and functional organization of genes for resistance to potato cyst nematodes are given. The results of molecular research on revealing the polymorphisms of loci involved in the control of resistance to cyst and gall nematodes, the development of molecular markers of certain genes and their use in marker-assisted selection for developing of new resistant cultivars, including those with group resistance, are considered.

[Antioxidant and membranotropic action of monochromanes and trimethylphenols derivatives in vitro].

Using some biochemical and physico-chemical methods of investigation, antioxidant activity and membrane protective effect of derivatives of monochromanes and trimethylphenols has been studied most active inhibitors of lipoperoxidation and the compounds capacity to structural modification of properties of the surface and hydrophobic zones of phospholipids bilayer in membranes of endoplasmic reticulum of the liver cells of rats have been determined.

Mapping of major spot-type and net-type net-blotch resistance genes in the Ethiopian barley line CI 9819.

Net blotch of barley (Hordeum vulgare L.), caused by the fungal phytopathogen Pyrenophora teres Drechs. f. teres Smedeg., constitutes one of the most serious constraints to barley production worldwide. Two forms of the disease, the net form, caused by P. teres f. teres, and the spot form, caused by P. teres f. maculata, are differentiated by the type of symptoms on leaves. Several barley lines with major gene resistance to net blotch have been identified. Earlier, one of these was mapped in the Rolfi x CI 9819 cross to barley chromosome 6H, using a mixture of 4 Finnish isolates of P. teres f. teres. In this study, we used the same barley progeny to map resistance to 4 spot-type isolates and 4 net-type isolates of P. teres. With all net-type isolates, a major resistance gene was located on chromosome 6H, in the same position as described previously, explaining up to 88% of the phenotypic variation in infection response in the progeny. We designate this gene Rpt5. Several minor resistance genes were located on chromosomes 1H, 2H, 3H, 5H, and 7H. These minor genes were not genuinely isolate-specific, but their effect varied among isolates and experiments. When the spot-type isolates were used for infection, a major isolate-specific resistance gene was located on chromosome 5H, close to microsatellite marker HVLEU, explaining up to 84% of the phenotypic variation in infection response in the progeny. We designate this gene Rpt6. No minor gene effects were detected in spot-type isolates. The Ethiopian 2-rowed barley line CI 9819 thus carries at least 2 independent major genes for net-blotch resistance: Rpt5, active against net-type isolates; and Rpt6, active against specific spot-type isolates.

[Genetic control of virulence of Pyrenophora teres drechs, the causative agent of net blotch in barley].

The genetic control of virulence was studied in four isolates of the fungus Pyrenophora teres f. teres, originating from various geographic regions in experiments with nine barley accessions, possessing known resistance genes. Experiments were performed with the ascospore progeny of two crosses. The results of segregation for virulence in the progeny of direct crosses were confirmed by analysis of backcrosses and sib crosses. One to four genes for avirulence toward various barley genotypes were found in the isolates under study. It is suggested that dominant suppressor genes are involved in the genetic control of avirulence toward four barley genotypes.