Stages Of Flower Bud Development Research Articles

Plant hormone signaling is involved in regulating flower bud size of daylily

To explore the role and metabolic differences of plant hormones in regulating the flower bud size of daylily, we collected the flower buds from two daylily varieties 'Datong Huanghua' and 'Dongbei Huanghua' at the young bud, green, yellowing, and yellow stages for transcriptome sequencing. The differentially expressed genes (DEGs) were screened, and the exogenous plant hormone spraying experiments were conducted. A total of 199 DEGs related to the biosynthesis and metabolism of plant hormones was screened out at different flower development stages of 'Datong Huanghua' and 'Dongbei Huanghua'. These genes regulated the biosynthesis and metabolism of six plant hormones: abscisic acid, gibberellin, auxin, jasmonate, cytokinin, and ethylene. The DEGs associated with auxin were the most, which suggested that auxin played a role in regulating flower bud development. The auxin response factor (ARF) presented up-regulated expression at all the four stages of flower bud development, indicating that ARF played a positive regulatory role throughout the flower bud development of daylily. The experiments with exogenous spraying of six hormones further verified that indole-3-acetic acid (IAA) significantly promoted the growth and increased the nutrient content in the flower buds of 'Datong Huanghua', suggesting that IAA played a role in regulating flower bud development. Our results laid a theoretical foundation for probing into the regulatory mechanism of flower bud development of 'Datong Huanghua' and 'Dongbei Huanghua'.

Characterization of Plant Homeodomain Transcription Factor Genes Involved in Flower Development and Multiple Abiotic Stress Response in Pepper.

Plant homeodomain (PHD) transcription factor genes are involved in plant development and in a plant's response to stress. However, there are few reports about this gene family in peppers (Capsicum annuum L.). In this study, the pepper inbred line "Zunla-1" was used as the reference genome, and a total of 43 PHD genes were identified, and systematic analysis was performed to study the chromosomal location, evolutionary relationship, gene structure, domains, and upstream cis-regulatory elements of the CaPHD genes. The fewest CaPHD genes were located on chromosome 4, while the most were on chromosome 3. Genes with similar gene structures and domains were clustered together. Expression analysis showed that the expression of CaPHD genes was quite different in different tissues and in response to various stress treatments. The expression of CaPHD17 was different in the early stage of flower bud development in the near-isogenic cytoplasmic male-sterile inbred and the maintainer inbred lines. It is speculated that this gene is involved in the development of male sterility in pepper. CaPHD37 was significantly upregulated in leaves and roots after heat stress, and it is speculated that CaPHD37 plays an important role in tolerating heat stress in pepper; in addition, CaPHD9, CaPHD10, CaPHD11, CaPHD17, CaPHD19, CaPHD20, and CaPHD43 were not sensitive to abiotic stress or hormonal factors. This study will provide the basis for further research into the function of CaPHD genes in plant development and responses to abiotic stresses and hormones.

A global view of transcriptome dynamics during flower development in Capsicum annuum L.

An analysis of genome-wide gene expression profiles during floral organ development can provide important clues about the molecular basis of gene functions and developmental processes. In this study, we analyzed the transcriptome data of 36 samples obtained during floral organ development from pepper ‘6421’ and detected 30 016 genes that were expressed in at least one sample. K-means clustering analysis was used to classify the data into 16 clusters based on the similarities between the dynamic expression profiles of genes. Of these, 15 clusters exhibited notable up-regulation or down-regulation trends in different developmental stages or tissues of floral organs. We identified transcription factors expressed at the early, medium, and late stages of bud development (F1, F5, F9). Transcription factor families such as AP2-ERF, MADS-box, MYB, bHLH, and NAC showed significant levels of enrichment. In comparison with genes expressed in vegetative tissues at different stages, certain genes were specifically up-regulated during flower development; among these, the number of genes specifically up-regulated during the stamen (Sta10) and bud tetrad development (F4) stages was the highest. Through extensive studies of the ABCDE model of flower development in Arabidopsis, we identified 17 ABCDE model candidate genes in pepper, most of which were up-regulated at specific stages of flower bud development. The expression data provided in this study is the most comprehensive dataset available for pepper to date and will serve as a resource for identifying the functions of many specific genes involved in flower development in pepper and other Solanaceae plants.

Industrial chicory genome gives insights into the molecular timetable of anther development and male sterility.

Industrial chicory (Cichorium intybus var. sativum) is a biannual crop mostly cultivated for extraction of inulin, a fructose polymer used as a dietary fiber. F1 hybrid breeding is a promising breeding strategy in chicory but relies on stable male sterile lines to prevent self-pollination. Here, we report the assembly and annotation of a new industrial chicory reference genome. Additionally, we performed RNA-Seq on subsequent stages of flower bud development of a fertile line and two cytoplasmic male sterile (CMS) clones. Comparison of fertile and CMS flower bud transcriptomes combined with morphological microscopic analysis of anthers, provided a molecular understanding of anther development and identified key genes in a range of underlying processes, including tapetum development, sink establishment, pollen wall development and anther dehiscence. We also described the role of phytohormones in the regulation of these processes under normal fertile flower bud development. In parallel, we evaluated which processes are disturbed in CMS clones and could contribute to the male sterile phenotype. Taken together, this study provides a state-of-the-art industrial chicory reference genome, an annotated and curated candidate gene set related to anther development and male sterility as well as a detailed molecular timetable of flower bud development in fertile and CMS lines.

Candidate genes screening based on phenotypic observation and transcriptome analysis for double flower of Prunus mume

BackgroundPrunus mume is an early spring flower of Rosaceae, which owns high application value in gardens. Being an excellent ornamental trait, the double flower trait has always been one of the important breeding goals of plant breeders. However, the key regulatory genes of double flower traits of P. mume are still unclear at present.ResultsThe floral organs’ morphological differences of 20 single and 20 double flower cultivars of P. mume were compared firstly. And it was found that double flower trait of P. mume were often accompanied by petaloid stamen, multiple carpels and an increase in the total number of floral organs. Then, transcriptome sequencing of two representative cultivars P. mume ‘Danban Lve’ and P. mume ‘Xiao Lve’ were conducted at 3 Stage of flower bud development with distinct morphological differentiation. 3256 differentially expression genes (DEGs) were detected, and 20 candidate genes for double flower trait of P. mume were screened out including hub genes PmAP1–1 and PmAG-2 based on DEGs function analysis and WGCNA analysis. And it was found that epigenetic and hormone related genes may also play an important role in the process of double flower.ConclusionsThis study suggested that the double flower trait of P.mume is more like accumulation origin based on morphological observation. 20 genes and co-expression network related to the formation of double flower P. mume were preliminarily screened through transcriptomics analysis. The results provided a reference for further understanding of the molecular mechanism of double flower trait in P. mume.

Identification and Characterization of Trihelix Transcription Factors and Expression Changes during Flower Development in Pineapple

Trihelix transcription factors with helix–loop–helix–loop–helix characteristics are essential in plant growth, development, and response to adversity. Several potential functions of Trihelix have been studied in multiple plant species, but little is known about the features and functions of Trihelix genes in pineapple. In this study, 23 Trihelix members were identified and classified into 5 groups and were unevenly distributed in 17 chromosomes of pineapple. The results revealed that six fragment duplication events and one tandem duplication event were found through gene duplication analysis. Moreover, 4, 10, 21, and 23 homologous gene pairs were found between pineapple and Arabidopsis, grape, banana, and rice, respectively. The promoters of Trihelix have many cis-elements, especially in light and hormone response. The expression characteristics of AcTrihelix members showed obvious tissue specificities in different tissues. The expressions of AcTrihelix3, AcTrihelix8, AcTrihelix16, AcTrihelix19, AcTrihelix20, and AcTrihelix23 were maintained at high levels during the late stage of flower bud development. In floral organs, the expression of different members was very different. In conclusion, some AcTrihelix members may play important roles during the floral development of pineapple, and they provide resources for further studies of the function of Trihelix and the molecular mechanism during pineapple flower formation.

Transcriptome analysis of flower bud identified genes associated with pistil abortions between long branches and spur twigs in apricots (Prunus armeniaca L.).

Pistil abortions of flower buds occur frequently in many apricot cultivars, especially in long branches. However, the molecular mechanism underlying pistil abortion in apricots remains unclear. To better understand the molecular mechanism of pistil abortions between long branches and spur twigs, paraffin sections and high-throughput sequencing technology were employed to analyze the expression patterns of genes associated with pistil abortions during later flower bud development stage in ‘Shajinhong’ apricot. The result of stage III (separation of bud scales) was the critical stage of pistil abortion in apricots. A total of 163 differentially expressed genes were identified as candidate genes related to pistil abortion in long branches. These genes are implicated in programmed cell death, hormone signaling, cell wall degeneration, and the carbohydrate metabolism pathway. The results showed that the up-regulation of gene expression of Xyloglucan endotransglucosylase/hydrolase and β-glucosidase in flower buds might be the direct cause of cell wall breakdown and pistil necrosis in long branches. We hypothesize that there is a molecular relationship between pistil abortion before blooming and cellulose degradation, and then carbohydrate transport in the case of carbon deficiency in long branches. Our work provides new insights into cellulose degradation in abortion pistils and valuable information on flower development in apricots, and also provides a useful reference for cultivation regulation in apricot or other fruit crops.

Genome-wide Identification, Characterization and Expression profile of TALE gene family in (Juglans regia L.)

The three amino acid loop extension (TALE) are essential transcription factors that regulate the differentiation of meristem in plant growth and development. Several studies on the TALE family in plants, including Arabidopsis, rice, and others, have been carried out. However, little is known about the TALE family in walnut. In this study, the genome-wide identification and expression analysis of the TALE gene family was studied comprehensively in walnut. Based on the specific structure of the TALE domain, a total of thirty walnut TALE genes were identified. According to different domains structure and phylogenetic relationships, the JrTALE gene family could be divided into two subfamilies, namely BELL and KNOX. Conserved protein motifs analysis showed that JrTALE contained similar motif structures in the same phylogenetic branch. The JrTALE gene family members had a weak overall codon bias, preferring to use synonymous codons ending in A/U. Expression pattern analysis of the JrTALE family genes showed that JrTALE17 and JrTALE18 were differentially expressed in female and male flower buds. We confirmed that JrTALE3, JrTALE5, JrTALE7, JrTALE10, JrTALE15 and JrTALE22 were differentially expressed between different stages of flower bud development by real-time quantitative PCR (qRT-PCR). This study provides valuable information about the TALE family in the transitional development of walnut flower buds, laying the foundation for a better understanding of the TALE family in walnut.

Biological and economic evaluation of AMISTAR Gold fungicide against sunflower rust in the Lower Volga region

Rust is one of the most economically important foliar diseases of sunflower. The problem of sunflower disease control was considered. The article presents the results of a two-year experiment studying the efficiency of AMISTAR Gold fungicide, concentrated suspension (active ingredient: Azoxystrobin 125 g/L, Difenoconazole 125 g/L) in sunflower hybrids in the Lower Volga region. Flowerbud development stage was the best application timing for increasing productivity. Therefore, sunflower yield averaged 34 c/ha. The biological efficiency of the fungicide ranged from 85 to 90 %. The calculation of economic efficiency showed 145255 % profitability depending on the year.

Abscisic acid mediates the reduction of petunia flower size at elevated temperatures due to reduced cell division.

Elevated temperatures suppress cell division in developing petunia buds leading to smaller flowers, mediated by ABA. Flower size is one of the most important showy traits in determining pollinator attraction, and a central factor determining the quality of floricultural products. Whereas the adverse effects of elevated temperatures on showy traits have been described in detail, its underlining mechanisms is poorly understood. Here, we investigated the physiological mechanism responsible for the reduction of flower size in petunia under elevated temperatures. We found that the early stages of flower-bud development were most sensitive to elevated temperatures, resulting in a drastic reduction of flower diameter that was almost independent of flower load. We demonstrated that the temperature-mediated flower size reduction occurred due to a shorter growth period, and a lower rate of corolla cell division. Consistently, local application of cytokinin, a phytohormone that promotes cell division, resulted in recovery of flower dimensions when grown under elevated temperatures. Hormone analysis of temperature-inhibited flower buds revealed no significant changes in levels of cytokinin, and a specific increase of abscisic acid (ABA) levels, known to inhibit cell division. Moreover, local application of ABA on flower buds caused a reduction of flower dimensions as a result of lower levels of cell division, suggesting that ABA mediates the reduction of flower size at elevated temperatures. Taken together, our results shed light on the mechanism by which elevated temperatures decrease petunia flower size, and show that temperature-mediated reduction of flower size can be alleviated by increasing the cytokinin/ABA ratio.

Morphological characteristics and transcriptome analysis at different anther development stages of the male sterile mutant MS7\u20132 in Wucai (Brassica campestris L.)

BackgroundThe discovery of male sterile materials is of great significance for the development of plant fertility research. Wucai (Brassica campestris L. ssp. chinensis var. rosularis Tsen) is a variety of non-heading Chinese cabbage. There are few studies on the male sterility of wucai, and the mechanism of male sterility is not clear. In this study, the male sterile mutant MS7–2 and the wild-type fertile plant MF7–2 were studied.ResultsPhenotypic characteristics and cytological analysis showed that MS7–2 abortion occurred at the tetrad period. The content of related sugars in the flower buds of MS7–2 was significantly lower than that of MF7–2, and a large amount of reactive oxygen species (ROS) was accumulated. Through transcriptome sequencing of MS7–2 and MF7–2 flower buds at three different developmental stages (a–c), 2865, 3847, and 4981 differentially expressed genes were identified in MS7–2 at the flower bud development stage, stage c, and stage e, respectively, compared with MF7–2. Many of these genes were enriched in carbohydrate metabolism, phenylpropanoid metabolism, and oxidative phosphorylation, and most of them were down-regulated in MS7–2. The down-regulation of genes involved in carbohydrate and secondary metabolite synthesis as well as the accumulation of ROS in MS7–2 led to pollen abortion in MS7–2.ConclusionsThis study helps elucidate the mechanism of anther abortion in wucai, providing a basis for further research on the molecular regulatory mechanisms of male sterility and the screening and cloning of key genes in wucai.

Foliar Application of Low Concentrations of Titanium Dioxide and Zinc Oxide Nanoparticles to the Common Sunflower under Field Conditions.

Nano-fertilisers have only recently been introduced to intensify plant production, and there still remains inadequate scientific knowledge on their plant-related effects. This paper therefore compares the effects of two nano-fertilisers on common sunflower production under field conditions. The benefits arising from the foliar application of micronutrient-based zinc oxide fertiliser were compared with those from the titanium dioxide plant-growth enhancer. Both the zinc oxide (ZnO) and titanium dioxide (TiO2) were delivered by foliar application in nano-size at a concentration of 2.6 mg·L−1. The foliar-applied nanoparticles (NPs) had good crystallinity and a mean size distribution under 30 nm. There were significant differences between these two experimental treatments in the leaf surfaces’ trichomes diversity, ratio, width, and length at the flower-bud development stage. Somewhat surprisingly, our results established that the ZnO-NPs treatment induced generally better sunflower physiological responses, while the TiO2-NPs primarily affected quantitative and nutritional parameters such as oil content and changed sunflower physiology to early maturation. There were no differences detected in titanium or zinc translocation or accumulation in the fully ripe sunflower seeds compared to the experimental controls, and our positive results therefore encourage further nano-fertiliser research.

MawuAP1 promotes flowering and fruit development in the basal angiosperm Magnolia wufengensis (Magnoliaceae).

The APETALA1/SQUAMOSA (AP1/SQUA)-like genes of flowering plants play crucial roles in the development processes of floral meristems, sepals, petals and fruits. Although many of the AP1/SQUA-like genes have been characterized in angiosperms, few have been identified in basal angiosperm taxa. Therefore, the functional evolution of the AP1/SQUA subfamily is still unclear. We characterized an AP1 homolog, MawuAP1, from Magnolia wufengensis that is an ornamental woody plant belonging to the basal angiosperms. Gene sequence and phylogenetic analyses suggested that MawuAP1 was clustered with the FUL-like homologous genes of basal angiosperms and had FUL motif and paleoAP1 motif domain, but it did not have the euAP1 motif domain of core eudicots. Expression pattern analysis showed that MawuAP1 was highly expressed in vegetative and floral organs, particularly in the early stage of flower bud development and pre-anthesis. Protein-protein interaction pattern analysis revealed that MawuAP1 has interaction with an A-class gene (MawuAP1), C-class gene (MawuAG-1) and E-class gene (MawuAGL9) of the MADS-box family genes. Ectopic expression in Arabidopsis thaliana indicated that MawuAP1 could significantly promote flowering and fruit development, but it could not restore the sepal and petal formation of ap1 mutants. These results demonstrated that there are functional differences in the specification of sepal and petal floral organs and development of fruits among the AP1/SQUA-like genes, and functional conservation in the regulation of floral meristem. These findings provide strong evidence for the important functions of MawuAP1 in floral meristem determination, promoting flowering and fruit development, and further highlight the importance of AP1/SQUA subfamily in biological evolution and diversity.

Biological characteristics and cytological studies on anther abortion of male sterile <italic>Camellia crassocolumna</italic>

The purpose of this study was to explore the flower morphology, anther and pollen abortion time and its cytological characteristics in male sterile plants of C. crassocolumna . The flowering process, floral morphology, anther structure, meiosis and microsporogenesis of male sterile (M350) and fertile plants (M352) of C. crassocolumna were observed by stereomicroscopy, paraffin section, chromosome preparation and DAPI staining. The results showed that the wild tea plant had a perfect flower, its anthers were tetrasporangiate shaped like a butterfly, with an anther wall development of basic type. It tapetal cells were binuclear, forming secretory cells at the tetrad stage, and degrading during mononuclear pollen stage. After the meiosis I, meiosis II and cytokinesis, pollen mother cells developed into tetrahedral tetrads. Microspores were triangular, and mature pollen was two-celled. The stamens of male sterile flowers developed normally in the early stage of flower bud development, and there was no difference between sterile and fertile flowers, while in the late stage of flower bud development, filaments were curved, anthers adhered and shriveled, anthers could not release pollen. The tapetal cells of male sterile flowers were proliferation abnormally and disordly at the meiosis of stage pollen mother cells, and the degradation of tapetum was delayed during the mononuclear and binucleate pollen periods. During meiosis of pollen mother cells of male sterile flowers, there were some abnormal appearances, such as chromosome ring, lagging chromosomes, chromosome bridges, chromosomal deletion, scattered chromosomes, unequal separation, micronucleus and abnormal tetrad. In sterile plants, the microspore cytoplasm was disorder, pollen grains adhered to each other at the mononuclear stage, pollen wall crumpled and deformed, cytoplasm and nucleus of pollen were blurred, the mature pollen cells were hollow and sunken. Taken together, the floral organ morphology of male sterile plants in C. crassocolumna belongs to stamen collapse type and anther abnormality type. The development of anthers is blocked from meiosis of pollen mother cells to uninucleate pollen stage, belonging to pollen mother cell abortion type and uninucleate pollen abortion type. Mononuclear period is the main period of anther abortion. Tapetum degradation delayed, chromosomal abnormalities of pollen mother cells in meiosis, and abnormal development of microspores and pollens, might be the main reasons for anther abortion in male sterile plants of C. crassocolumna .

Essential oil composition of Artemisia abrotanum L. during different vegetation stages in Lithuania

Artemisia abrotanum L. was introduced in the Middle of Lithuania Collection of Spice–Melliferous Plants of the Scientific Sector of Medicinal and Aromatic Plants of the Botanical Garden ex situ at Vytautas Magnus University since 1980. The object of investigation was Artemisiae abrotani herba of Artemisia abrotanum L. All samples were collected at different vegetation stages: growth and leaf production, flower bud development, the beginning of flowering, massive flowering and the end of flowering. Essential oils were obtained by hydrodistillation. The chemical composition of the essential oils of Artemisiae abrotani herba was studied by GC/MS in the Department of Analytical and Toxicological Chemistry at the Lithuanian University of Health Science in 2018–2019. Fifty-six compounds of the oil were identified, of which (+)-piperitone was the major component (20.38–38.48%). The data in this study showed a remarkable quantitative variation of constituents in the essential oils. The highest content and diversity of compounds was determined during the flower bud development stage. Sixty identified compounds in this stage reached 76.6% of the identified oil content. The highest concentration of this compound (38.48%) was detected at the end of the flowering vegetation stage and the lowest amount (20.38%) was found during the growth and leaf production. Among the other major compounds were (+)-piperitone, 1,4-cineole, lavandulyl butyrate, aromandendrene and isogermacrene D.

Honeybush (Cyclopia spp.) pollen viability and surface morphology

Genotypes of Cyclopia, used to produce honeybush tea, are being improved through selective breeding for suitability in large-scale cultivation and consistent flavour/aroma qualities. Though high levels of natural fecundity have been reported in honeybush species, attempts at controlled crosses have hitherto resulted in virtually no seed set. While investigating this problem, it was found that many basic questions regarding the specific reproductive physiology of the genus have not been answered. Studies were undertaken to provide the necessary data about sexual reproduction in honeybush species, in support of existing breeding programmes. This paper presents data concerning pollen viability in relation to flower bud stage, storage and pollen surface morphology of C. longifolia, C. maculata and C. subternata. Pollen anthesis and viability (measured as in vitro pollen germination percentage - PGP11Pollen germination percentage) were linked to visual stages of flower bud development. Pollen samples, dried and stored for 540 days at -18 °C, were tested for PGP at intervals (7, 30, 90, 180, 365, 540 days) during the storage period. While there was an overall decrease in PGP over storage time, four genotypes did not have any significant decrease in PGP over time. The effect of media sucrose content (MSC22Media sucrose content) (0–25%) on in vitro PGP of honeybush pollen was tested and differences in response were observed between species. Pollen surface morphology was investigated using FE-SEM microscopy and no consistent differences in shape or surface ornamentation were observed between the species, though differences in grain size were observed.

Reducing apple scab and frogeye or black rot infections with salicylic acid or its analogue on field-established apple trees

Apple scab (Venturia inaequalis) (AS) and frogeye leaf spot or black rot (Botryosphaeria obtusa) (FLS or BR) are two diseases of apples that affect developing foliage and fruit during summer and cause significant losses in the field and during storage. In this 3-year study (2014–2016), salicylic acid (2-hydroxybenzoic acid) or its analogue (Benzothidiazole or Actigard®) were applied to field-established apple ‘Honeycrisp’ at the tight cluster stage of flower bud development and at late pink bloom as either trunk injection or foliar sprays and to ‘Cortland’ (injection only). AS and FLS leaf infections were assessed 1, 2, and 3 weeks after the second treatment while infections on fruit were assessed at harvest. Main treatment effects were significant (P = 0.05) and both salicylic acid and Actigard® showed less infections on foliage due to AS in 2014 and 2016, and only salicylic acid showed less BR infections on fruit in both years. The cultivar effects were also significant, showing consistently less AS infections on the foliage and fruit of ‘Honeycrisp’, and less FLS infections on ‘Cortland’ foliage. Injection and spray treatments of salicylic acid and Actigard® showed no differences in infections by AS and FLS or BR on foliage or fruit. The method of application × treatment interactions were only significant in 2014; both salicylic acid and Actigard®-treated ‘Cortland’ trees showed less FLS infections on foliage and less BR infections on fruit, and only salicylic acid-treated ‘Honeycrisp’ trees showed less percentage of AS and BR infected fruit. In 2015, both salicylic acid and Actigard® treatments showed less infections of FLS on foliage and lower BR-infected fruit on ‘Cortland’. In 2016, both these treatments reduced incidence of AS and FLS on ‘Cortland’ foliage, and only salicylic acid reduced AS infections on ‘Honeycrisp’ foliage. Further studies are needed to integrate salicylic acid or Actigard® treatments into an apple foliar disease management program with registered fungicides.

Antioxidant Activity and Bioactive Compounds Contents in Different Stages of Flower Bud Development from Three Spanish Caper (<i>Capparis spinosa</i>) Cultivars

Antioxidant Activity and Bioactive Compounds Contents in Different Stages of Flower Bud Development from Three Spanish Caper (<i>Capparis spinosa</i>) Cultivars

Exploration of miRNA-mediated fertility regulation network of cytoplasmic male sterility during flower bud development in soybean.

Cytoplasmic male sterility (CMS) plays an important role in the production of soybean hybrid seeds. MicroRNAs (miRNAs) are a class of non-coding endogenous ~ 21nt small RNAs that play crucial roles in flower and pollen development by targeting genes in plants. To dissect the function of miRNAs in soybean CMS, a total of 558 known miRNAs, 10 novel miRNAs, and 466 target genes were identified in flower buds of the soybean CMS line NJCMS1A and its restorer line NJCMS1C through small RNA sequencing and degradome analysis. In addition, miRNA-mediated editing events were also observed, and the two most frequently observed editing types (A → G and C → U) were validated by cloning and sequencing. And as the base editing occurred, some targets were filtered, such as gma-miR2118b-P6GT with Glyma.08G122000.2. Further integrated analysis of transcriptome and small RNA found some miRNAs and their targets' expression patterns showing a negative correlation, such as gma-miR156b/GmSPL9a and gma-miR4413b/GmPPR. Furthermore, opposite expression pattern was observed between gma-miR156b and GmSPL9 during early stage of flower bud development. Taken together, the regulatory network of gma-miR156b/GmSPL9 and gma-miR4413b/GmPPR with flower bud development in soybean CMS was developed. Most importantly, previous reports showed that these targets might be related to pollen development and male sterility, suggesting that both conserved and species-specific miRNAs might act as regulators of flower bud development in soybean CMS. These findings may provide a better understanding of the miRNA-mediated regulatory networks of CMS mechanisms in soybean.

INFLUENCE OF SPRING FROST ON APPLE FLOWER BUDS AT VARIOUS DEVELOPMENTAL STAGES

Spring frost often reduces the yield because of damaged flowers. Spring frost is a dangerous climatic hazard that can be responsible for yield loses to orchard trees. Frost damage is highly dependent on the stage of development of the flower buds. Flowers buds samples were collected from ‘Granny Smith’, ‘Golden Delicious Rainders’ and ‘Gala Schniga’ apple cultivars at different flower buds stages from BBCH 59 (most flowers with petals forming a hollow ball) to BBCH 65 (full bloom on trees), two days after frost that occurred on the morning at 5:00-7:00 of March 31, 2017 when the temperature dropped to -1.5 to -3.3°C in the apple orchard of company Pollino Agrar near Fruška gora mountain. Pistils in flower buds samples at a hollow ball stage to full bloom were examined individually and classified as dead or alive based on tissue browning. The highest damage on flowers was recorded to cultivar ‘Gala Schniga’ (75.4%) and the lowest to cultivar ‘Golden Delicious Rainders’. Cultivar ‘Gala Schniga’ had more than 90% of damaged king flowers. Our results showed that the spring frost damage of pistils was variable according to stages of flower bud development and genotypes.