DISTRIBUTION OF SIVERS APPLES IN THE TERRITORY OF THE TARBAGATAI STATE NATIONAL NATURAL PARK

The current task in the study of Malus sieversii is the balanced use and protection, conservation, reforestation of the vegetation cover. Malus sieversii is a valuable gene pool on the territory of the State National Natural Park «Tarbagatai», therefore, the study, preservation of its form diversity is necessary for modern and future breeding, reforestation, afforestation. In order to preserve Malus sieversii in its original form, phenological monitoring of environmental, anthropogenic, natural and climatic factors of conditions affecting it is carried out and studied. An important role in increasing the productivity, quality and sustainability of Malus sieversii plantings is played by methods of conducting phenological monitoring control throughout the year, for many years.

Trichoderma-Induced Ethylene Responsive Factor MsERF105 Mediates Defense Responses in Malus sieversii

Trichoderma can induce plant hormone signal pathways mediating plant defenses, resulting in broad-spectrum resistance to phytopathogens. Herein, Malus sieversii seedlings were treated with Trichoderma biofertilizer and/or Alternaria alternata f. sp. mali, and transcriptome analysis revealed significant differential expression. There was a high similarity between the transcriptome expression profiles of Trichoderma-induced and A. alternata-infected M. sieversii samples for genes related to jasmonic acid (JA), ethylene, and salicylic acid (SA) signaling pathways. Additionally, Trichoderma biofertilizer activated numerous disease-resistant genes (ERF, NAC, bHLH, and STK) and defense response genes (DRP, ABC, and HSP). Among transcription factors, members of the ERF family were the most differentially expressed (18 ERFs), indicating that they may be closely related to defense responses. Among ERFs, differential expression of MsERF105 was the most significant (upregulated 27.6-fold compared to controls). MsERF105 was heterologously expressed in PdPap poplar (Populus davidiana × Populus alba var. pyramidalis Louche), and following infection with A. alternata (Aal), transgenic PdPap-MsERF105s plants displayed lower malondialdehyde (downregulated 41.4%) and reactive oxygen species (ROSs) levels, and higher reductase activities, especially superoxide dismutase (SOD; upregulated 77.5% compared to PdPap-ROK2 plants). Furthermore, the lesion areas of PdPap-MsERF105s leaves were significantly smaller (0.2%) than those of PdPap-ROK2 leaves (∼26.0%), and the cell membrane integrity was superior for PdPap-MsERF105s leaves. Thus, MsERF105 enhanced the resistance of PaPap poplar to Aal, presumably because MsERF105 activates the expression of PR1 and PDF1.2. In conclusion, Trichoderma biofertilizer modulated the differential expression of numerous disease resistance genes and defense response genes in M. sieversii in response to pathogen attack, and MsERF105 played important roles in this process.

Wild Apple-Associated Fungi and Bacteria Compete to Colonize the Larval Gut of an Invasive Wood-Borer Agrilus mali in Tianshan Forests

The gut microflora of insects plays important roles throughout their lives. Different foods and geographic locations change gut bacterial communities. The invasive wood-borer Agrilus mali causes extensive mortality of wild apple, Malus sieversii, which is considered a progenitor of all cultivated apples, in Tianshan forests. Recent analysis showed that the gut microbiota of larvae collected from Tianshan forests showed rich bacterial diversity but the absence of fungal species. In this study, we explored the antagonistic ability of the gut bacteria to address this absence of fungi in the larval gut. The results demonstrated that the gut bacteria were able to selectively inhibit wild apple tree-associated fungi. Among them, Pseudomonas synxantha showed strong antagonistic ability, producing antifungal compounds. Using different analytical methods, such as column chromatography, mass spectrometry, HPLC, and NMR, an antifungal compound, phenazine-1-carboxylic acid (PCA), was identified. Activity of the compound was determined by the minimum inhibitory concentration method and electron microscopy. Moreover, our study showed that the gut bacteria could originate from noninfested apple microflora during infestation. Overall, the results showed that in newly invaded locations, A. mali larvae changed their gut microbiota and adopted new gut bacteria that prevented fungal colonization in the gut.

New to Kazakhstan species of zygomycetes in forest soil of Trans-Ili Alatau and Kungey Alatau

This article provides data on two species of soil zygomycetes, new to Kazakhstan, found in the rhizosphere of various woody plants in the Trans-Ili and Kungey Alatau (Northern Tien Shan) – Piptocephalis cylindrospora and Lichtheimia corymbifera. P. cylindrospora is an obligate parasite of zygomycetous fungi; in the study area, it was found twice in Kungey Alatau on species Absidia spinosa and Lichtheimia corymbifera in the rhizosphere of Populus tremula. It does not cause a noticeable deterioration in the development of the host. Earlier, in Kazakhstan, only Piptocephalis arrhiza was registered in the rhizosphere of Armeniaca vulgaris, Populus tremula, Picea schrenkiana, Juniperus spp. Lichtheimia corymbifera, previously belonging to the genus Absidia, was recorded in the rhizosphere of Populus tremula in Kungey Alatau and in the rhizosphere of Picea schrenkiana, Salix sp., Crataegus sp. in Trans-Ili Alatau. L. corymbifera is characterized by heights of 1677 m in Kungey Alatau, and from 1516 to 2007 m a. s. l. in Trans-Ili Alatau. Until recently, in Kazakhstan, only a closely related species Absidia spinosa was isolated from the rhizosphere of Malus sieversii, Armeniaca vulgaris, Crataegus spp., Pinus sylvestris, Populus spp., Sorbus tianschanica, Picea schrenkiana, Salix spp. Descriptions of species colonies based on isolated pure cultures, morphological data, information on ecology and distribution are offered. In L. corymbiferacultures, the so-called “self-parasitism” is observed, in which the fungal hyphae colonize sporangiophores, and the latter look like penetrated by hyphae. The sexual stage of both species was not found in our studies.

Transcriptome profiling of Malus sieversii under freezing stress after being cold-acclimated

Background Freezing temperatures are an abiotic stress that has a serious impact on plant growth and development in temperate regions and even threatens plant survival. The wild apple tree (Malus sieversii) needs to undergo a cold acclimation process to enhance its freezing tolerance in winter. Changes that occur at the molecular level in response to low temperatures are poorly understood in wild apple trees. Results Phytohormone and physiology profiles and transcriptome analysis were used to elaborate on the dynamic response mechanism. We determined that JA, IAA, and ABA accumulated in the cold acclimation stage and decreased during freezing stress in response to freezing stress. To elucidate the molecular mechanisms of freezing stress after cold acclimation, we employed single molecular real-time (SMRT) and RNA-seq technologies to study genome-wide expression profiles in wild apple. Using the PacBio and Illumina platform, we obtained 20.79G subreads. These reads were assembled into 61,908 transcripts, and 24,716 differentially expressed transcripts were obtained. Among them, 4410 transcripts were differentially expressed during the whole process of freezing stress, and these were examined for enrichment via GO and KEGG analyses. Pathway analysis indicated that “plant hormone signal transduction”, “starch and sucrose metabolism”, “peroxisome” and “photosynthesis” might play a vital role in wild apple responses to freezing stress. Furthermore, the transcription factors DREB1/CBF, MYC2, WRKY70, WRKY71, MYB4 and MYB88 were strongly induced during the whole stress period. Conclusions Our study presents a global survey of the transcriptome profiles of wild apple trees in dynamic response to freezing stress after two days cold acclimation and provides insights into the molecular mechanisms of freezing adaptation of wild apple plants for the first time. The study also provides valuable information for further research on the antifreezing reaction mechanism and genetic improvement of M. sieversii after cold acclimation.

The Siberian wild apple, Malus baccata (L.) Borkh., is an additional contributor to the genomes of cultivated European and Chinese apples

It is crucial to understand domestication to unravel the evolutionary processes that shape the divergence of populations. Differences in life-history traits have probably led to marked differences in the mode and speed of evolution between trees and annuals, particularly the extent of crop-wild gene flow during domestication. Apple is an iconic tree and major fruit crop grown worldwide. The contribution of wild apple species to the genetic makeup of the cultivated apple genome remains a topic of intense investigations. We used population genomics in combination with SNPs to investigate the contributions of the two known wild apple relatives, Malus sylvestris and Malus sieversii, and a supposed contributor, Malus baccata, to European and Chinese rootstock and dessert genomes, with a focus on the extent of wild-crop gene flow during apple domestication. We showed that the European dessert and rootstock apples form a specific gene pool, whereas the Chinese dessert and rootstock apples were a mixture of three wild gene pools. Coalescent-based inferences and gene flow estimates indicated that M. baccata is an additional contributor to the genome of both European and Chinese cultivated apples through wild-to-crop introgressions. We also confirmed previous results on the contribution of M. sylvestris to the cultivated apple genome, and provided insights into the origin of the apple rootstock. This study further demonstrates the role of gene flow during apple domestication, as seen in other woody perennials, and show that domestication of the apple tree involved several wild apple species.

Ecosystems of protected area of the Natural Park “Medeu”

The article presents the results of mapping ecosystems of the protected area of the Natural Park “Medeu”,located on the northern slope of the Trans-Ili Alatau mountain range. The map of ecosystems and a legend has beencompiled. The legend contains 13 numbers. The largest sections of the legend are the natural ecosystems of the protectedarea and the natural-anthropogenic ecosystems of the buffer zone and the zone of limited economic activity. The habitatsof species from Red Data book of Kazakhstan that form communities (Malus sieversii (Ledeb.) M. Roem., Armeniacavulgaris Lam., Celtis australis subsp. caucasica (Willd.) C. C. Towns. identified, and their brief characteristics are given.

Genome-Wide Characterization of HSP90 Gene Family in Malus sieversii and Their Potential Roles in Response to Valsa mali Infection

Heat shock protein 90 (HSP90) is highly conservative molecular chaperon produced by plants in response to adverse environmental stresses including fungal infection. In China, canker disease, caused by Valsa mali, is the main threat for Malus sieversii, an ancestor of the cultivated apple. In this study, a total of eight HSP90 genes were identified from the M. sieversii genome and randomly distributed on eight chromosomes. According to gene structure and phylogenetic analysis, the MsHSP90s can be divided into five categories. The transcriptome analysis of M. sieversii under V. mali infection showed that the plant pathogen interaction pathway was identified as significantly enriched. RNA-seq data and qRT-PCR analysis demonstrated that the MsHSP90-6a gene was significantly repressed by V. mali infection. We further predicted cis-regulatory elements on the promotor region of MsHSP90 genes and identified canonical SHE motifs. Our results improve our understanding of the HSP90 gene family and provide a foundation for further studies of disease prevention in M. sieversii.

Expression of the Malus sieversii NF-YB21 Encoded Gene Confers Tolerance to Osmotic Stresses in Arabidopsis thaliana

Drought is the main environmental factor that limits the yield and quality of apples (Malus × domestica) grown in arid and semi-arid regions. Nuclear factor Ys (NF-Ys) are important transcription factors involved in the regulation of plant growth, development, and various stress responses. However, the function of NF-Y genes is poorly understood in apples. Here, we identified 43 NF-Y genes in the genome of apples and conducted an initial functional characterization of the apple NF-Y. Expression analysis of NF-Y members in M. sieversii revealed that a large number of NF-Ys were highly expressed in the roots compared with the leaves, and a large proportion of NF-Y genes responded to drought treatment. Furthermore, heterologous expression of MsNF-YB21, which was significantly upregulated by drought, led to a longer root length and, thus, conferred improved osmotic and salt tolerance in Arabidopsis. Moreover, the physiological analysis of MsNF-YB21 overexpression revealed enhanced antioxidant systems, including antioxidant enzymes and compatible solutes. In addition, genes encoding catalase (AtCAT2, AtCAT3), superoxide dismutase (AtFSD1, AtFSD3, AtCSD1), and peroxidase (AtPER12, AtPER42, AtPER47, AtPER51) showed upregulated expression in the MsNF-YB21 overexpression lines. These results for the MsNF-Y gene family provide useful information for future studies on NF-Ys in apples, and the functional analysis of MsNF-YB21 supports it as a potential target in the improvement of apple drought tolerance via biotechnological strategies.

Species composition of the mycobiota of the Sievers apple tree (Malus sieversii (Ledeb) M. Roem.) in Kazakhstan

The mycobiota of the Sievers apple tree on the territory of the fruit forests of Kazakhstan (Tarbagatai, Ketmen, Karatau, Altyn-Emel, Talassky, Zailiysky and Dzhungarsky Alatau ranges) includes 64 species of fungi from threedivisions. The most widely represented division is Ascomycota – 48 species, and for the genera Cytospora, Aspergillus,Alternaria, the largest number of species is noted (4 species, 3 and 3 species, respectively). The largest group of fungi of21 species causes damage to apple fruits. Fungus species causing apple tree trunk rot and wood rot are considered to bethe most harmful.