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Original Article • Braz. J. Biol. 85 • 2025 • https://doi.org/10.1590/1519-6984.287949 linkcopy

Open-access Distribution of powdery mildew resistance genes in the Kazakhstan apple cultivar collection

Distribuição de genes de resistência ao oídio na coleção de cultivares de maçã no Cazaquistão

AuthorshipSCIMAGO INSTITUTIONS RANKINGS

Abstract

The monogenic resistance to powdery mildew (caused by Podosphaera leucotricha) of apple trees is crucial when selecting them. DNA markers allow it to differentiate apple cultivars by individual resistance traits and determine prospective genotypes with high reliability. The presented research determines the results of molecular genetics and phytopathological analyses of apple cultivars on loci Pl-2, Pl-1, Pl-w, and Pl-d for reactions to powdery mildew. The molecular screening procedure involved using seven gene-specific markers (EM M01, EM M02, AT20-450, OPN18 SCAR, OPU02 SCAR, dr70F/dr339, and dr55F/dr336R). The resistance genes Pl-1, Pl-2, Pl-w, and Pl-d were amplified in 30 of the 34 research cultivars, with 8 Kazakh and 26 other country apple cultivars among the ones researched, according to the results of a molecular screening. Although resistance genes are still helpful for breeding, they are only recommended for use in extended pyramids of multiple resistant genes. Several cultivars are excellent candidates for additional breeding programs against powdery mildew and for pyramiding the Podosphaera leucotricha resistant genes in new cultivars.

Keywords:
apple; genetic marker; resistance genes; powdery mildew; Podosphaera leucotricha ; reactions

Resumo

A resistência monogênica ao oídio, causado por Podosphaera leucotricha, das macieiras é crucial na sua seleção. Marcadores de DNA permitem diferenciar cultivares de maçã por traços de resistência individual e determinar genótipos prospectivos com alta confiabilidade. A pesquisa apresentada determina os resultados de análises genéticas moleculares e fitopatológicas de cultivares de maçã nos loci Pl-2, Pl-1, Pl-w e Pl-d para reações ao oídio. O procedimento de triagem molecular envolveu sete marcadores específicos: EM M01, EM M02, AT20-450, OPN18 SCAR, OPU02 SCAR, dr70F/dr339 e dr55F/dr336R. Os genes de resistência Pl-1, Pl-2, Pl-w e Pl-d foram amplificados em 30 das 34 cultivares pesquisadas, sendo oito cultivares de maçã do Cazaquistão e 26 outras cultivares de outro país, entre as pesquisadas, segundo os resultados de uma triagem molecular. Embora os genes de resistência ainda sejam úteis para a reprodução, eles são recomendados apenas para uso em pirâmides estendidas de múltiplos genes resistentes. Várias cultivares são excelentes candidatas para programas de melhoramento adicionais contra o oídio e para a pirâmide dos genes resistentes a Podosphaera leucotricha em novas cultivares.

Palavras-chave:
macieira; marcador genético; gene de resistência; oídio; Podosphaera leucotricha ; reações

1. Introduction

Podosphaera leucotricha (Ellis & Everh.) E. S. Salmon, is the leading cause of apple powdery mildew (Gañán et al., 2020). The fruit shape, yield, and quality of fruits can be significantly affected by this fungal disease that affects the buds, leaves, new shoots, flowers, and apple fruits (Papp et al., 2016; Zhang et al., 2021). A white to greyish powdery patch on the leaves, new shoots, flowers, and young apple fruits are the primary symptoms of the pathogen, which eventually turns brown. Powdery mildew affects all plant above-ground parts, from leaves to fruits, resulting in significant fruit yield losses (Kоzlovskaya et al., 2018). Apple powdery mildew is included in the list of hazardous insects, plant diseases, and weeds of the Republic of Kazakhstan. Primary infection is caused by mycelium overwintering in vegetative tissues or infected flower buds, which actively colonises young developing shoots under favourable conditions. Subsequently, spores are formed on the mycelium, a secondary infection source (Radwan and Darwesh, 2018; Lyzhin and Savel’eva, 2021). The most crucial period of protection against fungal diseases is during the spread of primary infection: it is necessary to prevent the formation of sources of secondary infection. Epiphytotic years can cause 100% shoot damage on susceptible apple cultivars and up to 50-80% loss of apple marketable yield (Kоzlovskaya et al., 2018).

Powdery mildew is an issue for most modern apple cultivars, which means they must be treated with fungicides up to 15 times per growing season, depending on the growing area and climate conditions (Markussen et al., 1995). Fungicides are effective and protect apple trees in gardens and nurseries; however, chemical control requires significant labor and material resources. In this regard, among the current tasks of modern apple breeding is to improve varieties and rootstocks that are resistant/tolerant to Podosphaera leucotricha. Some apple cultivars and species are known to be powdery mildew resistant, including Pl-1 (Malus robusta), Pl-2 (Malus zumi), Pl-w ('White Angel'), Pl-d (Hybrid D12) and Pl-m (Hybrid MIS) (Knight and Alston, 1968; Visser et al., 1976; Dayton, 1977; Gallott et al., 1985; Simon and Weeden, 1991). The presence of molecular markers for powdery mildew resistance genes permits the identification of their sources and the purposeful inserting of these genes into newly created apple cultivars with a marker associated with selection.

Despite identifying multiple sources of resistance to the infection, searching for new gene sources is ongoing to provide a broad range of resistance genes to further improve new cultivars with pyramidal-resistant properties. It has been proven that single-gene resistance is not an effective way to achieve long-term resistance, as the powdery mildew has overcome most resistance genes used in apples to the present day at some point (Bus et al., 2010).

Several markers have been developed and tested to determine the sources of mildew resistance genes. For identification, a RAPD marker OPAT20-450 was developed, which was later converted into a SCAR marker AT20. AT20-SCAR is located at a distance of 4.5 cM from the gene Pl-1. The relatively close relationship of both markers makes them suitable for marker selection of mildew-resistant apple plants carrying the gene Pl-1 from M. robusta. The resistance and susceptible genotypes 'Fiesta' x E295-4 were used to calculate the map distances of markers EM M01 and EM M02, which were 4.6 and 6.4 cM from Pl-w, respectively (Evans and James, 2003; Markussen et al., 1995; Gardner et al., 1995; Gardiner et al., 2003). The EMDM01 was developed by converting the AFLP into a SCAR marker (James et al., 2004). At the distal end of Linkage Group 12, this marker can be found on 5 cM of the Pl-d resistance gene (Gelvonauskis and Gelvonauskiene, 2003). This gene's resistance level to apple powdery mildew is higher than in Pl-1 and Pl-2 (James et al., 2004). Additionally, it has been confirmed that Pl-2 resistance to powdery mildew (Jänsch et al., 2015; Chagné et al., 2019) is linked to single nucleotide polymorphisms (SNPs). Based on AFLP data, SCAR markers EM M01, EM M02, and EM DM01 were created for Pl-w and Pl-d (Evans and James, 2003; James et al., 2004). Microsatellite markers CH03C02 and CH01D03 were utilized to identify the Pl-d resistance gene. These markers have been successfully used for screening collections of apple cultivars in Germany (Höfer et al., 2021) and the Czech Republic (Patzak et al., 2011), as well as for research wild populations of Malus orientalis of Iran (Amirchakhmaghi et al., 2018). In Belarus, researchers have investigated the distribution of genes that protect against powdery mildew in commercial and local apple trees and wild species of the genus Malus (Suprun et al., 2016; Lyzhin and Savel’eva, 2021). However, apples' rootstocks have rarely been studied using markers for mildew resistance genes.

DNA markers have also been identified for the apple powdery mildew resistance genes Pl-1 and Pl-2, mapped on chromosomes 12 and 11. These markers are currently being used in marker-assisted breeding programs for powdery mildew resistance (Dunemann et al., 2007; Markussen et al., 1995; Murdoch et al., 2003).

One important direction in the use of DNA markers to determine genes resistant to powdery mildew, along with the use of selective forms carrying target genes, is screening collections of genetic resources to study the nature of sustainability and identify the genitor of sought-after genes. Our research focused on identifying genes resistant to powdery mildew Pl-2, Pl-1, Pl-w, and Pl-d in some apple cultivars collected from commercial orchards in southern Kazakhstan.

2. Material and Methods

2.1. Plant material and survey

The study materials included 34 apple tree samples (Table 1) from industrial orchards in Kazakhstan's southern and southeast regions (Shamraj and Glushhenko, 2006). A phytopathological evaluation on powdery mildew disease was done in the orchard of PF “Khoram', situated in the Enbekshikazakh district of Almaty region. According to data from the leading agronomist PF, “Khoram,” the orchard has 34 apple cultivars. The orchard is 110 hectares in area, with an annual yield of 50 tons per hectare and a gross yield of 5500 tons. The seedlings are 5-7 years old, and the distance between rows is 2 meters. Each counting point was stopped during the phytosanitary assessment of fungal diseases. The control of each apple cultivar included between 95 and 100 trees. The infection on fruit tree trunks, leaves, and fruits was thoroughly examined. Three phytosanitary evaluations were carried out on powdery mildew from June 10 to July 30 between 2022 and 2024 growing years. It should be clearly stated that no fungicide application was made in the orchards examined. ‘Gala’, ‘Fuji’, ‘Golden Delicious’, ‘Quinte’, ‘Red Delicious’, ‘Divo’, ‘Korey’, ‘Stark’s Earliest’, ‘Summer Red’, ‘Starkrimson’, and ‘Idared’ are the common apple cultivars grown in this locations.

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Table 1
Results of PCR analysis and phytopathological evaluation of apple cultivars based on loci of multigenic resistance to powdery mildew.

At least 100 leaves from each tree were evaluated. The number of infected leaves was divided by the total number of leaves to estimate the disease percentage (Reuveni and Reuveni, 1995).

The severity of the disease as a percentage was assessed on a modified scale (0-5) of Reuveni and Reuveni (1995), where:

  • 0 = No mold colonies examined;

  • 1 = 1-10% of leaf surface infected;

  • 2 = More than 10-25% of leaf surface infected;

  • 3 = More than 25-50% of the infected leaf surface;

  • 4 = More than 50-75% of the infected leaf surface;

  • 5 = More than 75-100% of the infected leaf surface.

2.2. DNA extraction and PCR

DNA was isolated from fresh green young leaves of different apple cultivars collected from survey areas using the CTAB protocol (Doyle and Doyle, 1987) with minor modifications (Madenova et al., 2024). Before isolation, the collected samples were frozen and stored at -80 °C. CTAB, as a detergent to disintegrate membranes, allows for the separation of DNA from proteins and lipids. During extraction, PVP is utilized to bind polyphenols. Cells are weakened by the chelating agent EDTA because it binds Mg++ and Ca++, which are vital for membrane stability. The DNA concentration in each sample was measured using agarose gel (1.5%) and later confirmed with Nanodrop (TM 2000/2000c ND-2000, Thermo Fisher Scientific Inc., Wilmington, DE, USA). Before the PCR, each sample was stored at -25 °C and had a final 5 ng/mL concentration.

The identification of Pl-1, Pl-2, Pl-d, and Pl-w resistance genes have been facilitated by the use of markers such as AT20-450, EM M02, EM M01, OPN18 SCAR, OPU02 SCAR, dr70F/dr339R, dr55F/dr336R in PCR reactions. The name, sequence, and annealing temperature of the primers are presented in Table 2. The reaction mixture for PCR contained 2 μL DNA, 1.25 μL MgCl2 buffer, 0.5 μL dNTP, 0.88 μL of each primer, 0.25 μL of Taq polymerase, 6.74 μL ddH2O, where the total was 12.5 μL. The amplification protocol was as follows: 94 °C - 2 min, 35 cycles 94 °C - 30 sec, T° an - 45 sec, 72 °C - 45 sec; 72 °C - 5 min (Evans and James, 2003).

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Table 2
Gene, marker name, primer sequence, and annealing temperature (°С) of primers used to identify genes for resistance to apple powdery mildew.

The amplified fragments were electrophoresed on a 1.5% agarose gel (w/v) using a 0.5-fold Tris/Borate/EDTA (TBE) buffer. The gel was dyed with ethidium bromide, and graphical images were obtained with the help of the UV Gel System (omniDOC SAFE Gel Documentation System; Cleaver Scientific Ltd, Rugby, United Kingdom). For measuring molecular mass, we used the DNA Ladder Plus gene which had a length of 50-100 bp.

The data analysis was conducted based on the molecular analysis results. The visualization of PCR products was accomplished through electrophoresis. Each extended PCR product had a band structure marked with '+' to indicate the presence, or '-' to indicate the absence of resistant genes. All the results of the data analysis are presented in Table 1 and Figure 1.

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Figure 1
The proportion of cultivars with different resistance to powdery mildew in the study collection is determined by field evaluation of resistance and molecular analysis.

3. Results

PCR analysis was carried out with 7 markers to determine genes resistant to the Podosphaera leucotricha. Molecular screening of 34 Kazakh and other country apple cultivars showed that 30 cultivars have genes resistant to powdery mildew. PCR analysis of apple genotypes was performed using the AT20-450 marker to identify carriers of the Pl-1 resistance gene (Figure 2 and Table 1). During the research, it was determined that 23 apple cultivars are carriers of the Pl-1 resistance gene: Talgar, Saltanat, Ainur, Zarya Alatau, Sinap Almaty, Gala, Red Delicious, Starkrimson, Fuji, Pink Lady, Scarlett, Red Chief, Vesna, Konfetnoe, Reinette Simirenko, White Transparent, Divo, Korey, Pestrushka, Williams Pride, Stark's Earliest and Landsberger Renette. Fragments that have a 450-500 length bp are detected due to amplification.

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Figure 2
Electrophoresis of marker AT20-450 with an amplified fragment length of 450-500 bp.

The EMM02 molecular markers were developed to align with the Pl-w resistance gene. However, PCR analysis did not reveal the Pl-w resistance gene in apple cultivars. The EM M01 marker was used to determine the stability resistance gene Pl-d. The amplification with the EM M01 marker determines fragments 90 bp long (Figure 3 and Table 1). The research determined that Sarkhyt, Saltanat, Talgar, Tyulpan, Ainur, Gala, Red Delicious, Granny Smith, Pink Lady, Jeromine, Vesna, Maminy Stakanchiki, Williams Pride, Quinte and Stark’s Earliest Pl-d resistance genes.

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Figure 3
Electrophoresis marker EM M01 with an amplified fragment length of 90 bp.

OPN18 SCAR, OPU02 SCAR, dr70F/dr339R, dr55F/dr336R have been developed for the Pl-2 resistance gene. As a result of PCR analysis with the help of markers dr70F/dr339R, the resistance gene Pl-2 was identified in 16 cultivars with fragments 270 bp long: Bala, Sinap Almaty, Red Delicious, Golden Delicious, Starkrimson, Babushkin, Granny Smith, Fuji, Pink Lady, Scarlett, Gala, Vesna, Konfetnoe, Maminy Stakanchiki, White Transparent, Landsberger Reinette (Figure 4 and Table 1). The dr55F/dr336R marker resulted in the detection of determined fragments measuring 290 bp (Figure 5 and Table 1). The gene Pl-2 was linked to 22 cultivars of apples using the molecular marker dr55F/dr336R. They are cultivars Saltanat, Danalyk, Talgar, Tyulpan, Zarya Alatau, Ainur, Sinap Almaty, Red Delicious, Golden Delicious, Starkrimson, Granny Smith, Pink Lady, Scarlett, Gala, Vesna, Konfetnoe, Maminy Stakanchiki, Reinette Simirenko, White Transparent, Korey, Pestrushka and Stark's Earliest.

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Figure 4
The amplification of dr70F/dr339R marker fragments resulted in the detection of lengths of 270 bp.
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Figure 5
Amplification with dr55F/ dr336R marker reveals fragments length 290 bp.

Amplification of the Pl-2 resistance gene with OPN18 SCAR and OPU02 SCAR markers revealed fragments of 620-650 bp and 1700 bp, respectively. PCR analysis did not identify the Pl-2 resistance gene based on OPN18 SCAR and OPU02 SCAR markers in apple cultivars.

The resistance of apple cultivars to powdery mildew was assessed between 2022 and 2024 growing years. The powdery mildew evaluation results clearly show that 13 cultivars have high resistance to the powdery mildew; Ainur, Tyulpan, Zarya Alatau, Starkrimson, Starkrimson, Granny Smith, Pink Lady, Vesna, White Transparent, Korey, Williams Pride, Quinte, Stark's Earliest and Landsberger Reinette. Nineteen cultivars were resistant to the powdery mildew with a reaction score of 1 (1-10% of leaf surface infected). These are the following: Bala, Sarkhyt, Saltanat, Danalyk, Talgar, Sinap Almaty, Red Delicious, Babushkin, Fuji, Scarlett, Jeromine, Gala, Red Chief, Idared, Konfetnoe, Maminy Stakanchiki, Reinette Simirenko, Divo and Pestrushka. The Golden Delicious and Aport cultivars, with two points for the disease, were moderately resistant (Figure 1, Table 1).

According to Table 1, most of the 34 apple cultivars are highly resistant to apple powdery mildew throughout tree year evaluation. This averages 55% to three years. Also, a small number of cultivars have moderate resistance to the Podosphaera leucotricha. Moreover, Figure 1 shows that most cultivars have the Pl-2 and Pl-1 genes in their genomes.

4. Discussion

To prevent economic losses caused by powdery mildew, it's best to use resistant apple cultivars. Determining and evaluating valuable sources of resistance in genetic resources is necessary for an effective resistance breeding process. The determination of resistance gene/genes and the development of resistant cultivars can be made easier with marker-assisted breeding (Iancu et al., 2023). Powdery mildew and scab are the most common fungal diseases that affect apple productions in Kazakhstan. The resistance genes to scab infection were determined in some commercial other country and Kazakh cultivars in our previous research (Madenova et al., 2024). In addition to fungal infections, there is also a chance of becoming infected by bacteria, such as bacterial fire blight caused by Erwinia amylovora (Ismailova et al., 2024). The research objective was to monitor the disease's spread and determine apple cultivars that exhibited resistance to powdery mildew.

In the phytopathological evaluation of cultivars from the Zhambyl region, three cultivars (Starkrimson, Golden Delicious, and Red Delicious) observed an immune reaction to powdery mildew. This is confirmed by molecular screening, where we see that 'Red Delicious' showed three genes (Pl-d, Pl-1, and Pl-2), and Starkrimson and Golden Delicious determined resistance genes Pl-1 and Pl-2.

A study conducted in Belarus reported that Malus baccatae has most significant number of Pl-1 marker alleles, with approximately 15 of the 19 cultivars (78.9%) exhibiting a recessive gene located at 450 bp. Additionally, the study determined various forms, including M. kirghisores, M. orientalis, M. prunifolia 2454, M. asiatica 2343, M. coronaria, M. florentina and M. sikkimensis (Lyzhin and Savel’eva, 2021). In this research, different cultivars have shown the presence of a 500 bp dominant gene. These include Red Delicious, Starkrimson, Granny Smith, Fuji, Pink Lady, Scarlett, Gala, Red Chief, Vesna, and Zarya Alatau, and the other 13 had a recessive gene.

M. baccatae apple trees and M. sieboldinae apple trees have the highest prevalence of the marker allele of the Pl-w resistance gene. In the M. baccatae series, the marker allele of the Pl-w resistance gene is found in 5 out of 19 forms, while in the M. sieboldinae series, it is determine in 4 out of 7 forms (Urbanovich et al., 2010). The marker EM M02 was identified in M. purpurea v. pendula 2396 in the Kyrgyz apple genotypes. The Pl-w resistance gene marker allele is absent in both eastern and native apple genotypes. The gene Pl-w resistance gene in M. sieboldii, M. sargentii 2428, and M. zumi has also been confirmed by other researchers (Urbanovich et al., 2010; Patzak et al., 2011). The same gene was not detected in any sampled orchards (Table 1).

PCR analysis of the Pl-2 resistance gene marker OPU02 SCAR in the cultivar ‘Favorite’ revealed a product of about 1500 bp. However, a fragment of 1700 bp is specific for the dominant allele of the Pl-2 resistance gene. At the same time, according to Suprun et al. (2015), it is known that the marker OPU02 SCAR gives amplification of a fragment of 2000 bp in the presence of the PlMIS powdery mildew resistance gene. From the results of this research, the Pl-2 powdery mildew resistance gene was not determined either through OPU02 SCAR or OPN18 SCAR marker in any of the cultivars. However, this gene was calculated using the marker systems dr55F/dr336R and dr70F/dr339R.

The analysis of the apple genome region that includes the gene Pl-2 revealed that a sequence of homologous stability genes belonging to the NBS-LRR class is closer to 4.9 cM from the gene (Gardiner et al., 2007). Based on this sequence, we selected convenient primers for amplification located at a shorter distance from the Pl-2 resistance gene. This research uses two pairs of markers to identify the Pl-2 resistance gene - dr70F/dr339R and dr55F/dr336R. The marker dr70F/dr339R showed up in 45.05% of the studied cultivars, among which only ‘Sinap Almaty’ was among the Kazakh cultivars, and the marker dr55F/dr336R showed up in 64.7% of the cultivars, among which Saltanat, Danalyk, Talgar, Tyulpan, Zarya Alatau, Ainur and Sinap Almaty were among the Kazakh cultivars.

To determine the Pl-d resistance gene, EMDM primer pairs were used to amplify the 90 bp region. Samples from ‘Ros1’, ‘GerdB4’, and ‘Challa’ contained this gene (James et al., 2004). James et al. (2004), identified the first set of molecular markers associated with the gene for mould resistance, Pl-d, using the bulked segregant method to test different microsatellites, amplified fragment length polymorphism (AFLP), and random amplified polymorphic DNA (RAPD) primers in germplasm separated from Pl-d (Radwan and Darwesh, 2018). This gene was detected in 44% of samples, particularly from Kazakhstan cultivars in Aport, Sarkhyt, Saltanat, Talgar, and Tyulpan.

It's important to note that species like Red Delicious, Granny Smith, Pink Lady, Gala, and Vesna have three resistance genes on AT20-450, EM M01, dr70F/ dr339R and dr55F/ dr336R, which means they have a genome resistant to mildew. Furthermore, the phytopathological analysis confirms the molecular screening data and supports this conclusion.

5. Conclusion

An examination of polymorphism in apple cultivars on loci of multifaceted resistance to powdery mildew was done using molecular markers. This research used molecular screening to determine apple cultivars reaction to powdery mildew. Powdery mildew was monitored in three regions of Kazakhstan, and PCR analysis was performed to determine genes resistant to the powdery mildew. Among the Kazakh apple cultivars, markers AT20-450 and EM M01 have been determined in Saltanat and Talgar. The presence of 5 out of 34 cultivars in four (AT20-450, EM M01, dr70F/ dr339R and dr55F/ dr336R) marker systems and 10 in three marker systems indicates their high resistance reaction to the powdery mildew. Of the 34 cultivars, 30.26% determine the Pl-1 resistance gene, 19.74% determine the Pl-d resistance gene, and 50% determine the Pl-2 resistance gene. However, the Pl-w resistance gene wasn’t determined in all of them. Future work in this field should be focused on developing marker assistant breeding programs to achieve resistant genotypes to apple powdery mildew. In this way, the varieties obtained using the markers will provide an opportunity for a project on a larger scale, in collaboration with plant pathologists.

Acknowledgements

The authors would like to thank the Laboratory of Plant Microclonal Propagation of Kazakh National Agrarian Research University for assistance in conducting this study. Moreover, this work was supported by the Project of the Ministry of Science and Higher Education of the Republic of Kazakhstan by Grant AP13068068 “Molecular screening and selection of resistant cultivars of apple tree to fungal diseases using DNA-marker technology” (2022-2024).

References

  • AMIRCHAKHMAGHI, N., YOUSEFZADEH, H., HOSSEINPOUR, B., ESPAHBODI, K., ALDAGHI, M. and CORNILLE, A., 2018. First insight into genetic diversity and population structure of the Caucasian wild apple (Malus orientalis Uglitzk.) in the Hyrcanian Forest (Iran) and its resistance to apple scab and powdery mildew. Genetic Resources and Crop Evolution, vol. 65, no. 4, pp. 1255-1268. http://doi.org/10.1007/s10722-018-0611-z
    » http://doi.org/10.1007/s10722-018-0611-z
  • BUS, V.G., BASSETT, H.C., BOWATTE, D., CHAGNÉ, D., RANATUNGA, C.A., ULLUWISHEWA, D., WIEDOW, C. and GARDINER, S.E., 2010. Genome mapping of an apple scab, a powdery mildew and a woolly apple aphid resistance gene from open-pollinated Mildew Immune Selection. Tree Genetics & Genomes, vol. 6, no. 3, pp. 477-487. http://doi.org/10.1007/s11295-009-0265-2
    » http://doi.org/10.1007/s11295-009-0265-2
  • CHAGNÉ, D., VANDERZANDE, S., KIRK, C., PROFITT, N., WESKETT, R., GARDINER, S.E., PEACE, C.P., VOLZ, R.K. and BASSIL, N.V., 2019. Validation of SNP markers for fruit quality and disease resistance loci in apple (Malus× domestica Borkh.) using the OpenArray® platform. Horticulture Research, vol. 6, no. 1, pp. 30. http://doi.org/10.1038/s41438-018-0114-2 PMid:30854208.
    » http://doi.org/10.1038/s41438-018-0114-2
  • DAYTON, D.F., 1977. Genetic immunity to apple mildew incited by Podosphaera leucotricha 1. HortScience, vol. 12, no. 3, pp. 225-226. http://doi.org/10.21273/HORTSCI.12.3.225
    » http://doi.org/10.21273/HORTSCI.12.3.225
  • DOYLE, J.J. and DOYLE, J.L., 1987. A rapid DNA isolation procedure for small quantities of fresh leaf tissue. Phytochemical Bulletin, vol. 19, no. 1, pp. 11-15.
  • DUNEMANN, F., PEIL, A., URBANIETZ, A. and GARCIA‐LIBREROS, T., 2007. Mapping of the apple powdery mildew resistance gene Pl-1 and its genetic association with an NBS–LRR candidate resistance gene. Plant Breeding, vol. 126, no. 5, pp. 476-481. http://doi.org/10.1111/j.1439-0523.2007.01415.x
    » http://doi.org/10.1111/j.1439-0523.2007.01415.x
  • EVANS, K.M. and JAMES, C.M., 2003. Identification of SCAR markers linked to Pl-w mildew resistance in apple. Theoretical and Applied Genetics, vol. 106, no. 7, pp. 1178-1183. http://doi.org/10.1007/s00122-002-1147-2 PMid:12748767.
    » http://doi.org/10.1007/s00122-002-1147-2
  • GALLOTT, J.C., LAMB, R.C. and ALDWINCKLE, H.S., 1985. Resistance to powdery mildew from some small-fruited Malus cultivars. HortScience, vol. 20, no. 6, pp. 1085-1087. http://doi.org/10.21273/HORTSCI.20.6.1085
    » http://doi.org/10.21273/HORTSCI.20.6.1085
  • GAÑÁN, L., WHITE III, R.A., FRIESEN, M.L., PEEVER, T.L. and AMIRI, A., 2020. A genome resource for the apple powdery mildew pathogen Podosphaera leucotricha. Phytopathology, vol. 110, no. 11, pp. 1756-1758. http://doi.org/10.1094/PHYTO-05-20-0158-A PMid:32515644.
    » http://doi.org/10.1094/PHYTO-05-20-0158-A
  • GARDINER, S.E., BUS, V.G.M., RUSHOLME, R.L., CHAGNÉ, D. and RIKKERINK, E.H.A., 2007. Apple. In: C. KOLE, ed. Fruits and nuts. Berlin: Springer, pp. 1-62. Genome Mapping and Molecular Breeding in Plants, vol. 4.
  • GARDINER, S., BASSETT, S., BASSETT, H., LEGG, W., RUSHOLME, R., BUS, V., RANATUNGA, C., DUNEMANN, F. and URBANIETZ, A., 2003. Marker assisted selection for pl-1 powdery mildew resistance in apple-old markers for a new resistance gene? Acta Horticulturae, no. 663, pp. 757-762.
  • GARDNER, B.D., JOHNSON, J.A. and HOLLAND, J.F., 1995. Rapid sampling for rapid analysis: a new method for VOCs in drinking water. In: Proceedings of the 43rd ASMS Conference on Mass Spectrometry and Allied Topics, 21-26 May, Atlanta, GA, USA. East Lansing, MI: ASMS, 1411 p.
  • GELVONAUSKIS, B. and GELVONAUSKIENE, D., 2003. Inheritance of resistance to powdery mildew and apple blotch in progenies of scab-resistant apple cultivars. Biologija, vol. 1, pp. 73-76.
  • HÖFER, M., FLACHOWSKY, H., SCHRÖPFER, S. and PEIL, A., 2021. Evaluation of scab and mildew resistance in the gene bank collection of apples in Dresden-Pillnitz. Plants, vol. 10, no. 6, pp. 1227. http://doi.org/10.3390/plants10061227 PMid:34208651.
    » http://doi.org/10.3390/plants10061227
  • IANCU, A., MILITARU, M. and STURZEANU, M., 2023. Molecular characterization of Romanian apple cultivars for identification of scab and powdery mildew resistance genes. Acta Horticulturae, no. 1384, pp. 409-418. http://doi.org/10.17660/ActaHortic.2023.1384.52
    » http://doi.org/10.17660/ActaHortic.2023.1384.52
  • ISMAILOVA, E., SHEMSHURA, O., SADANOV, A., BAIMAKHANOVA, G., TURLYBAYEVA, Z., KULDYBAYEV, N., YELUBAYEVA, A., KOPZHASSAROV, B., ISSINA, Z., TEMRESHEV, I. and BEKNAZAROVA, Z., 2024. Monitoring studies of the occurrence of fire blight pathogen in Kazakhstan and identification of antagonistic microorganisms suppressing its development. Brazilian Journal of Biology = Revista Brasileira de Biologia, vol. 84, e285493. http://doi.org/10.1590/1519-6984.285493 PMid:39109731.
    » http://doi.org/10.1590/1519-6984.285493
  • JAMES, C.M., CLARKE, J.B. and EVANS, K.M., 2004. Identification of molecular markers linked to the mildew resistance gene Pl-d in apple. Theoretical and Applied Genetics, vol. 110, no. 1, pp. 175-181. http://doi.org/10.1007/s00122-004-1836-0 PMid:15551035.
    » http://doi.org/10.1007/s00122-004-1836-0
  • JÄNSCH, M., BROGGINI, G.A., WEGER, J., BUS, V.G., GARDINER, S.E., BASSETT, H. and PATOCCHI, A., 2015. Identification of SNPs linked to eight apple disease resistance loci. Molecular Breeding, vol. 35, no. 1, pp. 45. http://doi.org/10.1007/s11032-015-0242-4
    » http://doi.org/10.1007/s11032-015-0242-4
  • KNIGHT, R.L. and ALSTON, F.H., 1968. Sources of field immunity to mildew (Podosphaera leucotricha) in apple. Canadian Journal of Genetics and Cytology, vol. 10, no. 2, pp. 294-298. http://doi.org/10.1139/g68-041
    » http://doi.org/10.1139/g68-041
  • KОZLOVSKAYA, Z.A., KONDRATSENOK, J.G., GASHENKО, T.A. and YARMOLICH, S.A., 2018. Identification of new complex sources of resistance to apple diseases using molecular methods in Belarus. Sadovodstvo i Vinogradarstvo, vol. 5, no. 5, pp. 23-29. In Russian. http://doi.org/10.31676/0235-2591-2018-5-23-29
    » http://doi.org/10.31676/0235-2591-2018-5-23-29
  • LYZHIN, A.S. and SAVEL’EVA, N.A., 2021. Polymorphism of wild-growing species of the genus Malus Mill for powdery mildew resistance genes. Vescì Nacyânal’naj Akadèmìì Navuk Belarusì. Seryâ Agrarnyh Navuk, vol. 1, no. 59, pp. 62-70. In Russian. http://doi.org/10.29235/1817-7204-2021-59-1-62-70
    » http://doi.org/10.29235/1817-7204-2021-59-1-62-70
  • MADENOVA, A., AITYMBET, Z., BOLAT, M., KALDYBAYEVA, D., GALYMBEK, K., KUAN, A., KABYLBEKOVA, B., IRKITBAY, A., YESZHANOV, T., BAKIROV, S. and SAPAKHOVA, Z., 2024. Screening of apple cultivars for scab resistance in Kazakhstan. Horticulturae, vol. 10, no. 2, pp. 184. http://doi.org/10.3390/horticulturae10020184
    » http://doi.org/10.3390/horticulturae10020184
  • MARKUSSEN, T., KRÜGER, J., SCHMIDT, H. and DUNEMANN, F., 1995. Identification of PCR‐based markers linked to the powdery‐mildew‐resistance gene Pl-1 from Malus robusta in cultivated apple. Plant Breeding, vol. 114, no. 6, pp. 530-534. http://doi.org/10.1111/j.1439-0523.1995.tb00850.x
    » http://doi.org/10.1111/j.1439-0523.1995.tb00850.x
  • MURDOCH, J., MEECH, S., RUSHOLME, R., BASSETT, H., COOK, M., BUS, V., RIKKERINK, E., GLEAVE, A., CROWHURST, R., ROSS, G. and WARRINGTON, I., 2003. Candidate resistance genes from an EST database prove a rich source of markers for major genes conferring resistance to important apple pests and diseases. Acta Horticulturae, vol. 622, pp. 141-151.
  • PAPP, D., KIRÁLY, I. and TÓTH, M., 2016. Suitability of old apple varieties in organic farming, based on their resistance against apple scab and powdery mildew. Organic Agriculture, vol. 6, no. 3, pp. 183-189. http://doi.org/10.1007/s13165-015-0126-2
    » http://doi.org/10.1007/s13165-015-0126-2
  • PATZAK, J., PAPRŠTEIN, F. and HENYCHOVÁ, A., 2011. Identification of apple scab and powdery mildew resistance genes in Czech apple (Malus× domestica) genetic resources by PCR molecular markers. Czech Journal of Genetics and Plant Breeding, vol. 47, no. 4, pp. 156-165. http://doi.org/10.17221/140/2011-CJGPB
    » http://doi.org/10.17221/140/2011-CJGPB
  • RADWAN, M.A. and DARWESH, D.R., 2018. Effect of integrated control program of powdery mildew disease on growth and productivity of apple. Journal of Plant Protection and Pathology, vol. 9, no. 12, pp. 787-794. http://doi.org/10.21608/jppp.2018.44066
    » http://doi.org/10.21608/jppp.2018.44066
  • REUVENI, M. and REUVENI, R., 1995. Efficacy of foliar sprays of phosphates in controlling powdery mildews in field-grown nectarine, mango trees and grapevines. Crop Protection, vol. 14, no. 4, pp. 311-314. http://doi.org/10.1016/0261-2194(94)00009-W
    » http://doi.org/10.1016/0261-2194(94)00009-W
  • SHAMRAJ, S.N. and GLUSHHENKO, V.I., 2006. The basics of field research in phytopathology and phytoimmunology: teaching and methodological manual Kharkiv: V. N. Karazin, 64 p.
  • SHAMSHIN, I.N., DUBROVSKY, M.L., TRIFONOVA, A.A., BORIS, K.V. and KUDRYAVTSEV, A.M., 2023. Powdery mildew resistance of apple clonal rootstocks from the collection of the Michurinsk State Agrarian University. Vavilovskii Zhurnal Genetiki i Selektsii, vol. 27, no. 6, pp. 572-581. http://doi.org/10.18699/VJGB-23-69 PMid:38023812.
    » http://doi.org/10.18699/VJGB-23-69
  • SIMON, C.J. and WEEDEN, N.F., 1991. Elucidation of crabapple lineage by direct examination of rDNA sequences. Malus, vol. 5, pp. 4-6.
  • SUPRUN, I.I., NASONOV, A.I., YAKUBA, G.V., LOBODINA, E.V. and BARSUKOVA, O.N., 2016. Effective selection of apple seedlings in a seed plot on resistance to scab and powder mildew. Fruit and Viticulture of the South of Russia, vol. 38, pp. 117-129. In Russian.
  • SUPRUN, I.I., TOKMAKOV, S.V., RISOVANNAJA, V.I., VOLODIN, V.A. and SHHERBATKO, V.D., 2015 [viewed 27 July 2024]. Identification of apples’ resistance genes to powdery mildew Pl-1 and Pl-2 and Vf and Vm for apple varieties from collections of genetic resources of the Crimean Peninsula. Scientific Works of the North Caucasus Zone Research Institute of Horticulture and Viticulture, vol. 7, pp. 15-19. In Russian. Available from: http://kubansad.ru/media/uploads/files/nauchnye_trudy_skzniisiv/tom_7/2.pdf
    » http://kubansad.ru/media/uploads/files/nauchnye_trudy_skzniisiv/tom_7/2.pdf
  • URBANOVICH, O.J., KOZLOVSKAYA, Z.A. and KARTEL, N.A., 2010. Distribution of powdery mildew resistant genes in the collection of varieties and apple species grown in Belarus. Molecular and Applied Genetics, vol. 11, pp. 20-25. In Russian.
  • VISSER, T., VERHAEGH, J.J. and DE VRIES, D.P., 1976. A comparison of apple and pear seedlings with reference to the juvenile period I. Seedling growth and yield. Euphytica, vol. 25, no. 1, pp. 343-351. http://doi.org/10.1007/BF00041566
    » http://doi.org/10.1007/BF00041566
  • ZHANG, Y., ZHANG, L., MA, H., ZHANG, Y., ZHANG, X., JI, M., VAN NOCKER, S., AHMAD, B., ZHAO, Z., WANG, X. and GAO, H., 2021. Overexpression of the apple (Malus× domestica) MdERF100 in Arabidopsis increases resistance to powdery mildew. International Journal of Molecular Sciences, vol. 22, no. 11, pp. 5713. http://doi.org/10.3390/ijms22115713 PMid:34071930.
    » http://doi.org/10.3390/ijms22115713

Publication Dates

  • Publication in this collection
    07 Feb 2025
  • Date of issue
    2025

History

  • Received
    27 July 2024
  • Accepted
    02 Dec 2024
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