Pollen Storage Research Articles

Blooming Phenograms, Pollen Production, and Pollen Quality during Storage of Pistachio Cultivars in New Mediterranean Growing Areas.

Pistachio (Pistacia vera L.) is a dioecious, anemophilous, and drought-resistant fruit tree species. It is cultivated in new Mediterranean areas, including the regions of southern Italy (Apulia and Basilicata). It has been estimated that over 40,000 ha are suitable for pistachio cultivation in areas infected by Xylella fastidiosa subsp. pauca. As a newly introduced species, knowledge of its biological reproductive behaviors in its new areas of spreading is essential for appropriate agronomic planning and management. This two-year study (2022 and 2023), carried out in the countryside of Stigliano (MT, Italy), had the objective of evaluating the flowering phenograms, pollen production, and assessing protocols for the conservation and extension of pollen viability, of the most widespread cultivars. A slight delay was observed in the blooming phenograms, compared to other cultivation Mediterranean areas, such as Spain or Sicily. Furthermore, the overlap between female and male phenograms was partial. No significant differences were observed in the polliniferous aptitude of the two male cultivars. Among the different protocols tested, the pollen storage at 33% relative humidity and a temperature of -80 °C maintained the pollen germinability above 50% for up to three weeks. These findings highlight the importance of controlled environmental conditions in preserving pollen viability over extended periods, providing valuable insights for agricultural and botanical research that relies on maintaining pollen viability for breeding and genetic studies.

Higher early than late-season residue load of pesticides in honey bee bread in Slovakia

Bees are often exposed to pesticide residues during their foraging trips in agricultural landscapes. The analysis of in-hive stored pollen reflects the spectrum of visited plants and can be almost used to link the exposure to pesticides.In the current study, bee bread samples were collected in May and July from 17 sites located in southern Slovakia. Samples were analysed using a multi-residue pesticide analysis method for a broad spectrum of active substances and microscopic for pollen identification.Our results revealed a bee bread contamination with 19 different active substances, with fungicides being predominant. Sixteen of them are authorized in the EU, but chlorpyrifos, chlorpyrifos-methyl, and chloridazon are not. The highest concentrations for pendimethalin (1400 µg/kg), fluazifop-butyl (640 µg/kg), fenpropidin (520 µg/kg), fluopyram (130 µg/kg), and difenoconazole (95 µg/kg) were detected. The total residue load in bee bread sampled in the early season (May) was significantly higher than in the late season (July). The mean residue load of insecticides analysed in July comprised 46% of May’s load, which is alarming due to the importance of bee bread in the diet for winter-rearing bees. Moreover, results from both sampling periods showed that fungicides were positively associated with plant families Apiaceae and Papaveraceae and herbicides with Aceraceae, Salicaceae, and Brassica-type/Brassicaceae.Hence, bee bread can be considered a suitable matrix and a good bio-indicator reflecting honey bee exposure to pesticides over the season.

Pollen products collected from honey bee hives experiencing minor stress have altered fungal communities and reduced antimicrobial properties.

Fungi are increasingly recognised to play diverse roles within honey bee hives, acting as pathogens, mutualists, and commensals. Pollen products, essential for hive nutrition, host significant fungal communities with potential protective and nutritional benefits. In this study, we profile the fungal communities and antifungal properties of three pollen products from healthy and stressed hives: fresh pollen collected by forager bees from local plants; stored pollen packed into the comb inside the hive; and bee bread, which is stored pollen following anaerobic fermentation used for bee and larval nutrition. Using amplicon sequencing, we found significant differences in fungal community composition, with hive health and sample type accounting for 8.8% and 19.3% of variation in beta diversity, respectively. Pollen and bee bread extracts had species-specific antimicrobial activity and inhibited the fungal hive pathogens Ascosphaera apis, Aspergillus flavus, and Aspergillus fumigatus, and the bacterial hive pathogen Paenibacillus larvae. Activity was positively correlated with phenolic and antioxidant content and was diminished in stressed hives. The plant source of pollen determined by amplicon sequencing differed in stressed hives, suggesting altered foraging behaviour. These findings illustrate the complex interplay between honey bees, fungal communities, and hive products, which should be considered in hive management and conservation.

Relative impacts of Varroadestructor (Mesostigmata:Varroidae)infestation and pesticide exposure on honey bee colony health and survival in a high-intensity corn and soybean producing region in northern Iowa.

The negative effects of Varroa and pesticides on colony health and survival are among the most important concerns to beekeepers. To compare the relative contribution of Varroa, pesticides, and interactions between them on honey bee colony performance and survival, a 2-year longitudinal study was performed in corn and soybean growing areas of Iowa. Varroa infestation and pesticide content in stored pollen were measured from 3 apiaries across a gradient of corn and soybean production areas and compared to measurements of colony health and survival. Colonies were not treated for Varroa the first year, but were treated the second year, leading to reduced Varroa infestation that was associated with larger honey bee populations, increased honey production, and higher colony survival. Pesticide detections were highest in areas with high-intensity corn and soybean production treated with conventional methods. Pesticide detections were positively associated with honey bee population size in May 2015 in the intermediate conventional (IC) and intermediate organic (IO)apiaries. Varroa populations across all apiaries in October 2015 were negatively correlated with miticide and chlorpyrifos detections. Miticide detections across all apiaries and neonicotinoid detections in the IC apiary in May 2015 were higher in colonies that survived. In July 2015, colony survival was positively associated with total pesticide detections in all apiaries and chlorpyrifos exposure in the IC and high conventional (HC) apiaries. This research suggests that Varroa are a major cause of reduced colony performance and increased colony losses, and honey bees are resilient upon low to moderate pesticide detections.

Fungal Diversity Associated with the Hive Stored Pollen of Stingless Bees Tetragonula travancorica Shanas and Faseeh

Fungal Diversity Associated with the Hive Stored Pollen of Stingless Bees Tetragonula travancorica Shanas and Faseeh

Molecular Identification and Prevalence of the Mite Carpoglyphus lactis (Acarina: Carpoglyphidae) in Apis mellifera in the Republic of Korea.

Apis mellifera, especially weak ones, are highly vulnerable to Carpoglyphus lactis mites, which can rapidly infest and consume stored pollen, leading to weakened colonies and potential colony collapse. This study aimed to ascertain and investigate the prevalence of this mite in honeybee colonies across nine provinces in the Republic of Korea (ROK). A total of 615 honeybee colony samples were collected from 66 apiaries during the spring and 58 apiaries during the summer of 2023. A 1242 bp segment of the Cytochrome c oxidase subunit 1 (COI) gene was amplified using the polymerase chain reaction method. The detection levels of C. lactis in the honeybees were compared between winter and summer. Based on the COI sequence analysis, the nucleotide sequence similarity of C. lactis mites isolated in the ROK with those from China (NC048990.1) was found to be 99.5%, and with those from the United Kingdom (KY922482.1) was 99.3%. This study is the first report of C. lactis in Korean apiaries. The findings of this study demonstrate a significantly higher detection rate in winter, which is 4.1 times greater than that in summer (p < 0.001). Furthermore, the results underscore the usefulness of molecular diagnostic techniques for detecting C. lactis mites.

Protective covers impact honey bee colony performance and access to outside resources

Protective crop covers are used extensively to protect high-value crops from bird and hail damage, however, these structures may present challenges for honey bees (Apis mellifera L.), the main pollinators in these systems. Some studies have indicated that protective covers result in declines in colony size and resource storage, but few replicated field trials exist. To determine the impact of protective cropping structures on honey bee brood production and resource storage, 14 honey bee colonies were placed inside and outside of protective covers on four blueberry (Vaccinium corymbosum and V. virgatum) farms in northern New South Wales, Australia. We measured changes in brood production, pollen and honey storage, hive weight and ambient hive temperature fortnightly over a 12-week period. Brood production, pollen and honey storage, and hive weight all displayed greater reductions within four weeks of placement under net cover, compared to hives outside of net cover. Although brood and honey metrics gradually increased in all hives during the remainder of the 12-week monitoring period, hives located under protective netting had relatively smaller gains in brood production and pollen storage when compared to hives located outside the protective covering. Hives under protective netting that were in the shade and/or whose entrances were facing south were most impacted by the end of the monitoring period. Our findings highlight the need for protected crop management strategies to consider pollinator health as well as crop pollination needs so that both bee health and pollination services are maintained in these systems.

Optimized Protocol for in vitro Pollen Germination in Brinjal (Solanum melongena L.)

Background: Understanding floral and pollen biology is essential for efficient artificial hybridization in hybrid seed production. This knowledge helps ensure proper fertilization of the ovary and optimal fruit and seed set. Selecting suitable pollen parents and preserving pollen for future use allows for better control and optimization of hybridization practices in brinjal breeding programs. To achieve this, a robust protocol for pollen collection and viability testing is necessary to monitor pollen quality in the field and during storage. This information is crucial for maintaining desired characteristics and genetic traits in the resulting hybrid seeds. Methods: This study; aimed at optimizing the pollen germination assessment protocol for brinjal. The step wise modification standard medium composition was done, the optimal growth condition was tested and in vivo predictions were made. So, the study aimed to standardization of media for in vitro pollen germination evaluate and viability in brinjal in ambient as well as cold storage conditions. The pollen germination was tested in vitro as well in vivo, by incubating the pollen grains for 3 hours at 25°C in different concentrations of sucrose, boron and agar media and distilled water at interval of 12 hours. Result: The most favorable germination of fresh pollen was achieved with a treatment consisting of 1% Agar, 12% Sucrose and 300 ppm Boron. Comparing the two storage conditions, pollen stored in the refrigerator exhibited higher in vitro germination percentages compared to ambient storage conditions. The highest in vitro pollen germination and viability percentages were observed in fresh pollen collected within 12 hours (95.00%). As the duration of pollen storage increased in both storage conditions, the in vitro pollen germination decreased. This decline in in vitro pollen germination was reflected in the in vivo fertilization fruit set percentage. The highest fruit set percentage was observed when pollen was stored in the refrigerator (64.52%) and when female flowers were pollinated with pollen stored for less than 12 hours (84.66%).

Standard methods to estimate strength parameters, flight activity, comb construction, and fitness of Apis mellifera colonies 2.0

This paper describes methods for estimating honey bee (Apis mellifera) colony strength by which mean population measures of adult bees and brood are obtained. Additionally, secondary measures, such as the quantity of stored honey and pollen, brood pattern, flight activity, comb construction, and the expression of visible disease or parasite symptoms are addressed. There are generally two contexts in which an investigator wishes to measure colony strength: (1) at the beginning of a study as part of manipulations to produce uniform colonies and reduce experimental error, and (2) as a response variable during or at the end of an experiment. Moreover, there are two general modes for measuring colony strength: (1) an objective mode that uses quantitative measures, and (2) a subjective mode that relies on visual estimates by one or more observers. Other parameters that do not directly measure colony strength are described because they give important indicators of colony state. These parameters include flight activity at the hive entrance, comb construction, and two proxy measures of colony fitness: production of queen cells and drone brood.

Gamma irradiation and ozone application as preservation methods for longer-term storage of bee pollen

Bee pollen is a healthy product with a good nutritional profile and therapeutic properties. Its high moisture content, however, promotes the growth of bacteria, molds, and yeast during storage commonly result in product degradation. Therefore, the aim of this study is to assess the effectiveness of gamma irradiation (GI) and ozone (OZ) as bee pollen preservation methods for longer storage time, as well as whether they are influenced by pollen species. To do that, GI at a dosage of 2.5, 5.0, and 7.5 kGy was applied at a rate of 0.68 kGy/h and OZ application at a concentration of 0.01, 0.02, and 0.03 g/m3 was applied for one time for 6 h, to Egyptian clover and maize bee pollen, then stored at ambient temperature for 6 months. We then determined the total phenolic content (TPC) and antioxidant activity of treated and non-treated pollen samples at 0, 3, and 6 months of storage. Total bacteria, mold, and yeast count were also evaluated at 0, 2, 4, and 6 months. Statistical analyses revealed that, TPC, antioxidant, and microbial load of both clover and maize pollen samples were significantly (p < 0.05) affected by both treatment and storage time and their interaction. Both methods were extremely effective at preserving the antioxidant properties of pollen samples after 6 months of storage at room temperature. Furthermore, the highest concentrations of both GI and OZ applications completely protected pollen samples from mold and yeast while decreasing bacterial contamination. GI at the highest dose (7.5 KGy) was found to be more effective than other GI doses and OZ application in preserving biologically active compounds and lowering the microbial count of pollen samples for 6 months. As a result, we advise beekeepers to use GI at this dose for longer-term storage.

Optimization of in vitro pollen storage conditions in seeded and low-seeded citrus genotypes

Optimization of in vitro pollen storage conditions in seeded and low-seeded citrus genotypes

Studies on pollen micro-morphology, pollen storage methods, and cross-compatibility among grape (Vitis spp.) genotypes.

The knowledge of pollen morphology, suitable storage condition, and species compatibility is vital for a successful grapevine improvement programme. Ten grape genotypes from three different species, viz., Vitis vinifera L., Vitis parviflora Roxb., and Vitis champini Planc., were studied for their pollen structure and pollen storage with the objective of determining their utilization in grape rootstock improvement programs. Pollen morphology was examined through the use of a scanning electron microscope (SEM). The viability of the pollen was assessed using 2,3,5-triphenyltetrazolium chloride (TTC). In vitro pollen germination was investigated using the semi-solid medium with 10 % sucrose, 100 mg/L boric acid, and 300 mg/L calcium nitrate. The results revealed variations in pollen micro-morphology in 10 genotypes, with distinct pollen dimensions, shapes, and exine ornamentation. However, species-wise, no clear difference was found for these parameters. Pollen of V. parviflora Roxb. and Dogridge was acolporated and did not germinate. The remaining eight genotypes exhibited tricolporated pollen and showed satisfactory in vitro pollen germination. Storage temperature and duration interactions showed that, at room temperature, pollen of most of the grape genotypes can be stored for up to 1 day only with an acceptable pollen germination rate (>30 %). However, storage for up to 7 days was successfully achieved at 4 °C, except for 'Pearl of Csaba'. The most effective storage conditions were found to be at -20 °C and -196 °C (in liquid N2), enabling pollen storage for a period of up to 30 days, and can be used for pollination to overcome the challenge of asynchronous flowering. Four interspecific combinations were studied for their compatibility, among which V. parviflora Roxb. × V. vinifera L. (Pusa Navrang) and V. parviflora Roxb. × V. champini Planc. (Salt Creek) showed high cross-compatibility, offering their potential use for grape rootstock breeding. However, V. parviflora Roxb. × V. vinifera L. (Male Hybrid) recorded the lowest compatibility index among studied crosses. In the case of self-pollinated flowers from V. parviflora Roxb. and V. parviflora Roxb. × V. champini Planc. (Dogridge), pollen failed to germinate on the stigma due to male sterility caused by acolporated pollen. As a result, the flowers of these genotypes functioned as females, which means they are ideal female parents for grape breeding without the need for the tedious process of emasculation.

Electrohydrodynamic drying of pollens based on a dual-polarization induction mode triboelectric nanogenerator

Drying pollen and other powders are important procedures of the food industry. Here we demonstrate a self-powered cold plasma drying system for collecting and drying pollen powders based on a dual-charge-induction mode triboelectric nanogenerator (TENG). The charge accumulation strategy of TENG has been modified by a dual-polarized module, where both ultra-high voltage output (over 30 kV) and sufficient output charge density could be generated to enhance drying efficiency. Comparing with all the other direct-current TENGs with an output voltage over 15 kV, this as-fabricated TENG can provide the highest transferred charge density so far. The electrohydrodynamic forces generated by the high-voltage TENG can be used to evaporate moisture from pollen. Within 20 min, it can lower the moisture content of fresh pollen from 35% to 7%, presenting a straightforward design for storing pollen at present. As for the same electrohydrodynamic drying process, the energy consumption of this self-powered TENG-based drying system for evaporating 1 kg water is about 5.36 kJ, which is only 36.4% of energy consumption of commercial high-voltage power source. This design provides a simple yet effective low-power consumption drying system for current pollen storage, which can be a promising environment-friendly alternative for food industry.

Pollen viability and longevity in Juniperus taxa native to Slovakia

Pollen viability, dispersion ability and longevity during deep-freezer storage were studied in three Juniperus taxa distributed in Slovakia. All these characteristics of pollen are closely related to the pollination and/or fertilization success of the junipers in nature. Pollen viability varied considerably between the three populations of J. communis and one population of each, J. sibirica and J. communis var. intermedia. Pollen germination rate ranged between 40.75% and 75.06%, and pollen tube length between 30.32 and 40.41 µm. A clear tendency indicates a higher germination rate of J. communis pollen from lower altitudes and reduced germination of J. sibirica and J. communis var. intermedia pollen from higher altitudes. The dispersion potential of the J. communis pollen during its shedding seems relatively low. In 2021, pollen cloud density was diluted at 68.1% at the 4 m distance from the test shrub, in 2022 of 52.1% at the 17 m distance from the pollen source. A deep-freezer storage of juniper pollen in a double-walled polyethylene bag with silica gel was not efficient enough, as indicated by the drop of pollen germination rate of 31.2% in J. communis and of 79.4% in J. sibirica during a 1-year storage period at − 81 °C.

Systemic pesticides in a solitary bee pollen food store affect larval development and increase pupal mortality

Solitary bees are often exposed to various pesticides applied for pest control on farmland while providing pollination services to food crops. Increasing evidence suggests that sublethal toxicity of agricultural pesticides affects solitary bees differently than the social bees used to determine regulatory thresholds, such as honey bees and bumblebees. Studies on solitary bees are challenging because of the difficulties in obtaining large numbers of eggs or young larvae for bioassays. Here we show the toxic and sublethal developmental effects of four widely used plant systemic pesticides on the Japanese orchard bee (Osmia cornifrons). Pollen food stores of this solitary bee were treated with different concentrations of three insecticides (acetamiprid, flonicamid, and sulfoxaflor) and a fungicide (dodine). Eggs were transplanted to the treated pollen and larvae were allowed to feed on the pollen stores after egg hatch. The effects of chronic ingestion of contaminated pollen were measured until adult eclosion. This year-long study revealed that chronic exposure to all tested pesticides delayed larval development and lowered larval and adult body weights. Additionally, exposure to the systemic fungicide resulted in abnormal larval defecation and increased mortality at the pupal stage, indicating potential risk to bees from fungicide exposure. These findings demonstrate potential threats to solitary bees from systemic insecticides and fungicides and will help in making policy decisions to mitigate these effects.

Sublethal effects of imidacloprid-contaminated honey stores on colony performance, queens, and worker activities in fall and early winter colonies.

Neonicotinoid-contaminated sugar stores can have both near term and long term effects on honey bees due to their persistence in honey stores. Effects of imidacloprid food stores contaminants were examined in subtropical colonies that experience reduced brood rearing and foraging during overwintering. Colonies were given treatment sugar syrup containing 0 ppb (control), 20 ppb (field relevant), or 100 ppb (above field relevant) imidacloprid over six weeks to simulate contaminated fall nectar. Colonies were evaluated immediately (post-treatment) and 10 weeks (mid-winter) after treatment to compare proximal and latent effects. Post-treatment 0 ppb and 20 ppb colonies had more workers than 100 ppb colonies while 0 ppb colonies more brood than 20 ppb or 100 ppb colonies. Mid-winter 0 ppb and 20 ppb colonies had more workers than 100 ppb colonies and 0 ppb colonies more brood than 100 ppb colonies. Colonies experienced seasonal declines in stored pollen but no treatment effects. Lower 100 ppb colony performance was associated with reduced effort rather than lifespan. RFID (Radio Frequency Identification) tracking revealed that workers had similar adult lifespans across treatments; however, 100 ppb workers engaged in activities outside the colony for less time than 0 ppb workers. Imidacloprid exposure affected queen but not worker nutritional physiology. Nurses retained well-developed hypopharyngeal glands (as indicated by head protein) across treatments. Mid-winter queens from 0 ppb colonies had marginally higher ovary protein than queens from 100 ppb colonies and more ovary lipids than queens from 20 ppb colonies. However, queen nutrient stores in non-reproductive tissues (fat bodies) did not differ across treatments. Queens from different treatments were attended by comparable numbers of retinue workers and had similar gland contents of four QMP (Queen Mandibular Pheromone) components essential to queen care. High levels of imidacloprid in sugar stores can negatively affect colony performance months after initial storage.

Review on Chalkbrood Disease of Honey Bee

Ascosphaera apis is the sole source of the invasive mycosis known as “chalkbrood disease”, which only affects honeybee larvae. Around 5-37% less honey is produced globally as a result of the illness, and 80% of broods perish. Because of the disease’s current severity, more pesticides are used, and colonies are frequently transported over great distances, which may have created new opportunities for Ascosphaera apis infection. There are signs that the incidence of this disease, which is now widespread worldwide, may be increasing. It is also highly unlikely that Ascosphaera apis will ever be completely eradicated due to its widespread distribution throughout the world, the fact that it can persist for more than 15-years in soil near infected apiaries, and the fact that viable spores can be found in stored honey, pollen, pollen capsules and tablets, used hive components, and used beekeeping tools and equipment. Safe control of the disease has been noted to be challenging, despite the fact that management techniques, medications, and the adoption of particular chalkbrood-resistant bees have all been demonstrated to have some benefits thus far. Thus, current research for a better understanding of chalkbrood will focus on genetic engineering for better fungus manipulation, molecular investigation of Ascosphaera apis pathogenesis pathways, and the potential identification of strategies to break the pathway through the identification of responsible genes for its pathogenesis. Some researchers have noted the prevalence of chalkbrood disease and linked chalkbrood diseases to bee colony losses and decreased productivity in affected bee colonies.

Effects of Short Term Pollen Storage on Fruit Set, Seed Number and Seed Germination Rate in Cut Rose Hybridization

Roses are known for their low pollen quality. It remains unclear how many days of pollen should be used in order not to reduce crossbreeding success. The study was conducted to determine whether short-term pollen storage is effective on the crossing success of roses comparatively in vivo and in vitro. The study was conducted in a greenhouse and a cytology laboratory at Ankara University, Turkey. The 'Avalanche' rose variety was used as the seed parent, and 'Magnum' was the pollen parent. A total of 210 crosses were made. Pollens were used immediately (day 0) and stored at +4°C for 6 days after collection. The pollen viability by IKI and the germination rates by the petri dish method, the fruit and seed set, the fruit and seed weight, and the seed germination rate were recorded. All examined traits showed a decreasing trend as the storage time increases. The pollen germination rates were recorded at 1.63% for stored pollen at six days and 15.27% for fresh pollen. The highest fruit and seed set rates were obtained from crossing with pollen stored for one day. The fruit set decreased below 10% as of the 5th day, and the seed numbers decreased 1.8 times compared to one day of storage. No germination was observed in seeds obtained from crosses with pollen stored for 5 or 6 days. A positive correlation was found between seed number and pollen germination rate. The study concluded that successful crossing could be made with pollen stored at +4°C for up to 3 days.

Metric and Spectral Insight into Bee-Pollen-to-Bee-Bread Transformation Process.

Due to numerous bioactive constituents, both bee pollen (BP) and bee bread (BB) represent valuable food supplements. The transformation of BP into BB is a complex biochemical in-hive process that enables the preservation of the pollen's nutritional value. The aim of this study was to determine the depth of the honeycomb cells in which bees store pollen and to provide a spectral insight into the chemical changes that occur during the BP-to-BB transformation process. This study was carried out on three experimental colonies of Apis mellifera carnica, from which fresh BP was collected using pollen traps, while BB samples were manually extracted from the cells two weeks after BP sampling. The samples were analyzed using infrared (FTIR-ATR) spectroscopy, and the depth of the cells was measured using a caliper. The results showed that the average depth of the cells was 11.0 mm, and that the bees stored BB up to an average of 7.85 mm, thus covering between ⅔ and ¾ (71.4%) of the cell. The FTIR-ATR analysis revealed unique spectral profiles of both BP and BB, indicating compositional changes primarily reflected in a higher water content and an altered composition of the carbohydrate fraction (and, to a lesser extent, the lipid fraction) in BB compared to BP.

Floral, reproductive, and pollination biology of Floral, reproductive, and pollination biology of Eugenia myrcianthes Nied.

Studies on species of the family Myrtaceae are mostly related to floristic surveys, reproduction involving large plant communities, and family taxonomy. Thus, the objective of this study was to elucidate aspects of the floral and reproductive biology, as well as floral visitors of ubajai (Eugenia myrcianthes Nied.). Studies were conducted on floral morphology and morphometry, identification of nectaries and structures attractive to floral visitors, characterization of floral visitors, stigma receptivity, and androecium maturation, pollen storage, in vitro viability testing, and characterization of the reproductive system. Ubajai flowers open at approximately 6:00 a.m., and their anthers are the main attractive structure to floral visitors. The main floral visitors and effective pollinators of ubajai are honeybees (Apis mellifera). The addition of 40% sucrose to the culture medium, using fresh pre-anthesis pollen, allows for a 90% germination rate. Ubajai pollen is recalcitrant, thus, it loses viability before 30 days of storage, even when stored in a refrigerator, freezer, liquid nitrogen, or natural environment. Ubajai can be considered a self-compatible plant; however, fertilization of flowers through cross-pollination also occurs, and apomixis does not occur.