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Average time trends (in days) in the viability of Cannabis sativa pollen grains. Statistically different values (p < 0.05) with each other.
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The aim of this study was to evaluate the pollen viability and longevity of Cannabis sativa, which is a significant economic crop. Viability and longevity are evaluated using different staining techniques: Alexander's, Triphenyl Tetrazolium Chloride (TCT) and Fluorochromatic Reaction (FCR). The results of this study show that FCR is the most reliab...
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... of results from days' data gives the general trend of pollen viability in all months that is greatest between 0-3 days of anther dehiscence, with the highest percentage on day 1. Viability days' trend in the month of April (which showed the highest viability percent- age: Figure 4) in which the maximum viability reported represented in Figure 3. The average percentage of via- bility in April lies between 70 and 78% on day 0, at the time of anthesis. ...
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This study was carried out on Euphrates River, which is one of the main sources of water in Iraq to assess the water quality of Southeast of DhiQar province. Water samples were collected monthly and seasonally at three stations from October 2016 to September 2017. Eight parameters were analyzed, water temperature, dissolved oxygen, total hardness,...
Citations
... In Poland, there are over ten plantain species, the most common of which are P. lanceolata, P. media, and P. major. Plantago major, a perennial herb, is well known for its medicinal properties and is used to treat a variety of ailments (Choudhary et al. 2014). This species is distinguished by leaves with a broadly rounded or cordate base, flower spikes with broadly round-ovate sepals, and ovoid fruits that split at or just below the center (Britannica 2020). ...
Aerobiology studies the biological particles in the air that affect human health, especially plant pollen that causes allergies. The purpose of this study is to describe and examine the pollen seasons of the following plants: plantain (Plantago sp.), mugwort (Artemisia sp.), alder (Alnus sp.), birch (Betula sp.), grasses (Poaceae sp.), and hazel (Corylus sp.) during the period 1996‒2021. The results show that each species experiences unique pollen seasons that are influenced by weather patterns. By compre-hending these patterns, pollen thresholds essential for controlling pollen allergies and reducing negative health effects can be established.
... In Poland, there are over ten plantain species, the most common of which are P. lanceolata, P. media, and P. major. Plantago major, a perennial herb, is well known for its medicinal properties and is used to treat a variety of ailments (Choudhary et al. 2014). This species is distinguished by leaves with a broadly rounded or cordate base, flower spikes with broadly round-ovate sepals, and ovoid fruits that split at or just below the center (Britannica 2020). ...
Aerobiology studies the biological particles in the air that affect human health, especially plant pollen that causes allergies. The purpose of this study is to describe and examine the pollen seasons of the following plants: plantain (Plantago sp.), mugwort (Artemisia sp.), alder (Alnus sp.), birch (Betula sp.), grasses (Poaceae sp.), and hazel (Corylus sp.) during the period 1996‒2021. The results show that each species experiences unique pollen seasons that are influenced by weather patterns. By comprehending these patterns, pollen thresholds essential for controlling pollen allergies and reducing negative health effects can be established.
... For longer day flight times, pollen viability may become a factor for risk of cross-pollination 15 . Choudhary et al. found that viability of Cannabis pollen only decreased substantially three days after release from the anther 33 . In our study, we are only considering dispersal within 50 km of the source. ...
... It is possible that hemp pollen only disperses during the day, as is common for many wind-dispersed species 27 . One study observed that male Cannabis anthers open and release pollen in the morning hours 33 . However, Cannabis pollen measurement studies found only slight diurnal changes in concentration 10,34 , indicating that Cannabis pollen remains in the air throughout the day. ...
For the recently legalized US hemp industry (Cannabis sativa), cross-pollination between neighboring fields has become a significant challenge, leading to contaminated seeds, reduced oil yields, and in some cases, mandated crop destruction. As a step towards assessing hemp cross-pollination risk, this study characterizes the seasonal and spatial patterns in windborne hemp pollen dispersal spanning the conterminous United States (CONUS). By leveraging meteorological data obtained through mesoscale model simulations, we have driven Lagrangian Stochastic models to simulate wind-borne hemp pollen dispersion across CONUS on a county-by-county basis for five months from July to November, encompassing the potential flowering season for industrial hemp. Our findings reveal that pollen deposition rates escalate from summer to autumn due to the reduction in convective activity during daytime and the increase in wind shear at night as the season progresses. We find diurnal variations in pollen dispersion: nighttime conditions favor deposition in proximity to the source, while daytime conditions facilitate broader dispersal albeit with reduced deposition rates. These shifting weather patterns give rise to specific regions of CONUS more vulnerable to hemp cross-pollination.
... Labor, expenses, and breeding danger are eliminated as a result. A crucial procedure for guaranteeing pollination, sustaining regular date palm bearing, particularly for early season flowering female cultivars, and conserving biodiversity is the storage of pollens while keeping increased viability and germinability (Choudhry et al., 2014). It is still unknown why wheat pollen has such a limited lifespan. ...
In technical breeding research and genetic resource conservation, the storage of vegetative plant components, including pollen, has become increasingly significant. Wheat pollen cannot sustain their viability for an extended period of time, in contrast to many other plant species.
... Acetocarmine stains chromatin located within the pollen nuclei, indicating pollen that is fully developed or has a complete structure [26]. In hemp, all three methods have been used in pollen quality investigations [21,22,27,28]; some are specifically used to compare pollen quality from different male flower stimulation methods [9]. ...
Female hemp plants are desired in floral hemp operations due to their higher cannabinoid contents. To produce feminized seeds, a critical step of inducing fertile male flowers on female plants is performed. In feminized seed production, freshly mixed STS (silver thiosulfate + sodium thiosulfate) is applied to female plants as an ethylene inhibitor to induce male flowers. However, the short-shelf stability of the STS buffer can cause difficulty in the application and inconsistent results. Alternative methods with improved accessibility and stable buffers will be beneficial for the hemp industry and hemp breeders. A commercially available floriculture product, Chrysal ALESCO®, contains silver nitrate, the same active ingredient as STS but with increased shelf stability. This study compares Chrysal ALESCO® to the traditional STS standard methods for male flower induction on female plants and their pollen quality. The two treatments were applied to six female hemp accessions with three replicates investigated, and the male flower counts and pollen quality were compared. No statistically significant difference was discovered in their male flower counts; the STS-treated plant produced an average of 478.18 male flowers, and the Chrysal ALESCO®-treated plant produced an average of 498.24 male flowers per plant. Fluorescein diacetate (FDA) and acetocarmine stains were used to investigate the pollen quality (non-aborted rate) of two chosen genotypes. FDA-stained pollen of Chrysal ALESCO® showed a significantly higher non-aborted rate than the pollen of traditional STS-treated plants (p < 0.001); however, only a marginally higher non-aborted rate was discovered by acetocarmine staining (p = 0.0892). In summary, Chrysal ALESCO® performed equally to traditional STS treatment at male flower counts and better or equally in pollen quality. With better shelf stability and easy application, ALESCO® can be a viable alternative option for stimulating male flowers on female hemp plants.
... Hemp is exclusively wind pollinated, although bees visit male flowers, they are not attracted to female flowers (Flicker et al. 2020). Pollen will quickly lose viability within three days at room temperature (Choudhary et al. 2014), but pollen may be stored for at least four months at liquid nitrogen temperature (Gaudet et al. 2020). ...
After decades of prohibition of the cultivation and breeding of hemp ( Cannabis sativa < 0.3% ∆9‐tetrahydrocannabinol [THC]), there is untapped potential for genetic improvement of this crop to provide food, feed, fiber, and medicinal compounds. Successful breeding efforts will require the development and characterization of germplasm resources, optimization of crossing methods, better understanding of sex determination, high‐throughput phenotyping platforms, and deployment of genomic tools for rapid selection. This review provides a brief overview of these topics and some key opportunities for genetic improvement of hemp to support an emerging industry utilizing this newly legalized crop.
... Other studies have shown that factors related to the in vitro culture medium can affect pollen germination (Flores-Rentería et al., 2018;Impe et al., 2020). In addition, there is information relating the reproductive potential of pollen grains to the time they are removed from the anther, which involves seasonality or floral development stadium (Choudhary et al., 2014). ...
... Flowers were collected from plants of the three accessions in pre-anthesis (a day before flower opening), at the day of the opening, and 72-h after flower opening, and sent to the Biotechnology laboratory. Pollen grains were carefully extracted from the anthers using a needle (0.45 × 13 mm, TKL brand) and inoculated in the center of a glass slide containing 150 μL of a standard culture medium (Brewbaker and Kwack, 1963): 1.27 mM Ca (NO 3 ) 2 ·4H 2 O, 0.87 mM MgSO 4 ·7H 2 O, 0.99 mM KNO 3 , 1.62 mM H 3 BO, and 100 g · L −1 sucrose. For each flower, three slides were prepared and kept in a BOD incubator at a temperature of 27 °C in the dark for 6-h (Soares et al., 2015). ...
... Flowers of the ICA-bd and ICA-vt accessions were collected on the first day of anthesis at 8 am. The pollen grains were extracted and inoculated as in second experiment, in adapted modified culture medium (Brewbaker and Kwack, 1963) with different concentrations of sucrose (0, 10%, 20%, and 30%), at pH 7.0. The slides, placed in Petri dishes with hydrated filter paper, were maintained in a BOD incubator at controlled temperatures of 15, 20, 25, and 30 °C for 6-h. ...
Adenium obesum (Forssk.) Roem. & Schult., popularly known as desert rose, has become a valuable ornamental plant. In floriculture, the production of hybrids is prioritized. Hence, knowledge on floral biology and sexual reproduction of the target species is fundamental. The objectives of this study were: (1) to test sucrose concentrations and temperatures for in vitro germination of A. obesum pollen grains; (2) to identify the effect of temperature on the viability of A. obesum pollen grains; and (3) to evaluate the viability of pollen grains and stigma receptivity in pre-anthesis, at flower opening, and 72 h post-flower opening in three accessions of A. obesum. A significant relationship between temperatures and sucrose concentrations was observed in the in vitro germination test. The highest percentage of in vitro germination of pollen grains, 39.81%, was observed at an estimated temperature of 26.05 °C. Desert rose accessions maintained in biochemical oxygen demand (BOD) chambers at 30 °C during a 16-h light photoperiod showed faster flowering, and temperatures ≥ 25 °C induced pollen grain viability percentages above 69%. Temperature is one of the most important abiotic factors, influencing mainly in pollen germination, pollen tube growing and in efficiency fertilization. The ICA-wd accession stood out and can be considered a pollen donor in artificial pollination. The stigmas of flowers were receptive from a day before flower opening until three days after. The two parameters presented above, stigma receptivity and pollen viability, allow inferences about the appropriate time for successful pollination and subsequent fertilization in desert roses. Resumo Viabilidade de grão de pólen e receptividade do estigma em Rosa-do-Deserto Adenium obesum (Forssk.) Roem. & Schult, conhecida popularmente como rosa-do-deserto, tem se tornado uma opção valiosa para o setor de plantas ornamentais. Na floricultura, a produção de híbridos é priorizada, para isso, o conhecimento da biologia floral e da reprodução sexuada da espécie é fundamental. Objetivou-se com o presente estudo: (1) testar concentrações de sacarose e temperaturas na germinação in vitro de grãos de pólen de Adenium obesum; (2) identificar o efeito da temperatura na viabilidade de grãos de pólen de Adenium obesum e; (3) avaliar a viabilidade dos grãos de pólen e receptividade estigmática pré-antese, abertura floral e 72 horas pós-abertura floral em três acessos de Adenium obesum. Interação significativa entre temperaturas e concentrações de sacarose foi observada no teste de germinação in vitro. A maior porcentagem de germinação in vitro dos grãos de pólen, 39,81% foi observada na temperatura estimada de 26,05 °C. Os acessos de rosa-do-deserto mantidos em B.O.D. sob temperatura de 30 °C e fotoperíodo de 16 horas de luz apresentaram florescimento mais rápido e as temperaturas ≥ 25 °C indicaram porcentagens de viabilidade dos grãos de pólen acima de 69%. A temperatura é um dos fatores abióticos mais importantes, influencia a germinação dos grãos de pólen, crescimento do tubo polínico e pode afetar a fertilização. O acesso ICA-bd destacou-se e pode ser considerado como doador de pólen em polinizações artificiais. O estigma das flores encontrou-se receptivo desde um dia antes da abertura das flores até três dias após a flor abrir. Os dois parâmetros apresentados acima, receptividade estigmática e viabilidade polínica, permitem fazer inferências importantes sobre os momentos mais apropriados para a polinização e subsequente fertilização na rosa-do-deserto. Palavras-chave: antese, floricultura, sacarose, temperatura. Cristiane Gonçalves souza et. al V. 28, N o. 1, 2022 p. 92-98 93
... Plant breeders store pollen to facilitate variety development and to preserve germplasm (Choudary et al., 2014). Hemp pollen is very delicate, and its ability to fertilize ovaries rapidly declines after anthesis. ...
... A variety of techniques to assess pollen viability have been developed, including staining pollen grains with fluorescein diacetate (FDA) to assess enzymatic activity, germinating grains on an agar-sucrose germination medium to observe pollen tube growth, or releasing airborne grains onto live receptive pistillate flowers to confirm seed set (Choudary et al., 2014;Gaudet et al., 2020). Assessing viability using FDA does not necessarily indicate the ability for a pollen grain to germinate. ...
... In the present study, pollen from masculinized female hemp plants had a minimal germination rate, initially and after storage, in contrast to the germination rates of pollen from genetically male plants (Choudary et al., 2014;Gaudet et al., 2020 Irregularly shaped pollen grains are commonly produced by masculinized hemp plants (DiMatteo et al., 2020) and, as a result, may negatively influence germination rate. ...
Cannabis sativa L. (hemp) develops plants with either male or female flowers, and growers of hemp greatly prefer female flowers which bear the glandular trichomes that contain cannabinoids. Feminized (all female) seeds are highly desired, which are produced by crossing a female plant with a masculinized female plant. Masculinization is achieved through the inhibition of ethylene and/or addition of gibberellins before flower initiation in female plants. The hemp industry uses silver thiosulfate (STS) to masculinize hemp, but spraying silver poses environmental concerns. This study compared STS to three other ethylene-inhibiting agents: aminoethoxyvinylglycine (AVG), cobalt nitrate (CBN), and 1-methylcyclopropene (1-MCP). Treatments of STS and CBN also included gibberellic acid as a synergist. Plants treated with STS exhibited superior masculinization and pollen dispersal compared to plants treated with AVG, CBN or 1-MCP. Only plants treated with STS or AVG produced pollen in sufficient quantities for collection. This pollen was assayed for germination potential initially and after storage for up to five weeks at 22.2, 7.2, or 1.1°C. Pollen from plants treated with AVG remained viable for four weeks at 1.1°C, whereas STS-treated plants produced pollen that was viable for three weeks at 1.1°C. Due to phytotoxicity problems with AVG, STS remains the best treatment to masculinize female hemp plants when breeding for feminized seeds. In a separate study, flower tissues of hemp had considerably higher total cannabinoid concentrations compared to leaf tissues but significantly lower ratios of cannabidivarin (CBDV) to cannabidiol (CBD). To reduce variability, at least 1 g samples of fresh leaf or flower tissue should be extracted with 10 mL of methanol. Rapid throughput testing of cannabinoids as part of a breeding program should use flower tissue, preferably at the time typical of harvest.
... The great amount of microspores and pollen grains produced in each flower bud (>150,000), together with the high rates of microspore viability reached for all phenotypes (>50%), makes C. sativa a suitable system able to provide, in a very quick period of time (≈30 days after seed germination), huge populations of viable vacuolate microspores and young bi-cellular pollen grains potentially inducible to afford microspore and pollen embryogenesis experiments. Viability rates obtained in our research are similar to the ones reported by Choudhary et al. (2014) which, depending on the season, oscillated among 33.3 and 83.0%, while in another work (Zottini et al., 1997) a viability rate for mature pollen grains of 92% was reported. This difference among viability rates could be attributable to the microspore isolation procedure, as viability measurements in the former works were performed on mature pollen grains naturally released after anthesis. ...
... As a result of a week-long cold pretreatment applied directly on excised buds prior to microspore culture, a stress-derived slight decrease in microspore viability was detected. Our findings are in agreement with results published by Choudhary et al. (2014), who studied how pollen viability was significantly influenced by seasonal fluctuations in temperature and humidity. Researchers reported a significant decrease of pollen viability at low temperatures, reaching the lowest viability rates in the winter season. ...
Development of double haploids is an elusive current breeding objective in Cannabis sativa L. We have studied the whole process of anther and pollen grain formation during meiosis, microsporogenesis, and microgametogenesis and correlated the different microgametophyte developmental stages with bud length in plants from varieties USO31 and Finola. We also studied microspore and pollen amyloplast content and studied the effect of a cold pretreatment to excised buds prior to microspore in vitro culture. Up to 476,903 microspores and pollen grains per male flower, with in vivo microspore viability rates from 53.71 to 70.88% were found. A high uniformity in the developmental stage of microspores and pollen grains contained in anthers was observed, and this allowed the identification of bud length intervals containing mostly vacuolate microspores and young bi-cellular pollen grains. The starch presence in C. sativa microspores and pollen grains follows a similar pattern to that observed in species recalcitrant to androgenesis. Although at a low frequency, cold-shock pretreatment applied on buds can deviate the naturally occurring gametophytic pathway toward an embryogenic development. This represents the first report concerning androgenesis induction in C. sativa, which lays the foundations for double haploid research in this species.
... Pollen viability is of great interest to breeders and researchers alike. Breeding projects may wish to store pollen for extended periods of time, where high value genetic material may be stored for future use or for biotechnological and gene editing applications that requires a quick and effective method for determining pollen viability [3][4][5]. Fluorescent stains such as fluorescein diacetate (FDA) or fluorochromatic reaction test (FCR) have been previously reported for assessing pollen viability in cannabis [3,4]. Viability is not always correlated with germination, as pollen may retain the ability to metabolize while losing its ability to germinate [5]. ...
... Breeding projects may wish to store pollen for extended periods of time, where high value genetic material may be stored for future use or for biotechnological and gene editing applications that requires a quick and effective method for determining pollen viability [3][4][5]. Fluorescent stains such as fluorescein diacetate (FDA) or fluorochromatic reaction test (FCR) have been previously reported for assessing pollen viability in cannabis [3,4]. Viability is not always correlated with germination, as pollen may retain the ability to metabolize while losing its ability to germinate [5]. ...
Pollen viability and storage is of great interest to cannabis breeders and researchers to maintain desirable germplasm for future use in breeding or for biotechnological and gene editing applications. Here, we report a simple and efficient cryopreservation method for long-term storage of Cannabis sativa pollen. Additionally, the bicellular nature of cannabis pollen was identified using DAPI (4′,6-diamidino-2-phenylindole) staining. A pollen germination assay was developed to assess cannabis pollen viability and used to demonstrate that pollen collected from different principal growth stages exhibited differential longevity. Finally, a simple and efficient method that employs pollen combined with baked whole wheat flour and subsequent desiccation under vacuum was developed for the long-term cryopreservation of C. sativa pollen. Using this method, pollen viability was maintained in liquid nitrogen after four months, suggesting long-term preservation of cannabis pollen.
