Effects of γ-ray treatment onCannabis saliva pollen viability

Abstract

The viability and the in vitro germination capability of hemp pollen (cv. Carmagnola) were studied. Viability tests were based on the microscopic observation of the fluorescence of loaded fluorescein diacetate (FDA), while, for germinability tests, five different media were tested. The effects of irradiation with 7-rays on pollen viability and germination and on seed set were also studied, at three different irradiation doses (20, 60 and 100 krad). The results show that in one of the media tested, about 85-90% of the pollen grains are viable and able to germinate in control samples, and that while viability measured by FDA test is not affected by increased 7-ray doses, the pollen in vitro germinability drops to about one-half of the controls at the maximum "y-ray dose employed, 100 krad. Seed set of hemp plants pollinated with the irradiated pollen dropped to less than 1% of that of plants pollinated by untreated pollen for the higher dose used. The different media suitable for in vitro germination of hemp pollen, and the observed lack of correspondence between viability and germination capacity tests are discussed.Abbreviations: FDA -fluorescein diacetate

Pollen Appearance and In Vitro Germination Varies for Five Strains of Female Hemp Masculinized Using Silver Thiosulfate

“…Irregular pollen grains could be the result of many things, including abnormal meiosis, improper exine formation, and problems with the tapetal cells (Baghali et al, 2011;Small, 1972). The pollen germination rates we observed were less than those reported for hemp by Zottini et al (1997) using the same germination media. This contrast may be attributable to differences in genotype and growing conditions.…”

“…This contrast may be attributable to differences in genotype and growing conditions. Zottini et al (1997) evaluated pollen germination for the fiber variety Carmagnola, which was grown outdoors in the field. Our pollen germination rates for CBD hemp are similar to those reported for five cultivars of tea hybrid rose (Voyiatzi, 1995).…”

“…Immediately after collection, pollen was spread on petri dishes containing a gelled medium using a soft bristled brush. The germination medium, described by Zottini et al (1997), was composed of 17% sucrose, 30 mg · L -1 Ca(NO 3 ) 2 , 100 mg · L -1 H 3 BO 3 , and 0.7% agar, with pH adjusted to 6.4. Three petri dishes were prepared per genotype.…”

Implications of ionizing radiation on pollen performance in comparison with diverse models of polar cell growth

“…All here presented data, which derive from the surveyed studies, were reconsidered and collectively standardized, and thus they are here comparatively indicated as percentage of their respective untreated controls (Tables 1, 2, & 3). In summary, IR caused a suppression of pollen germination (Table 1) and tube growth (Table 2) in a dose‐dependent manner (Cuny et al, 1993; Kumar & Rai, 2006; Peixe et al, 2000; Pfahler, 1971; Sadat Hosseini Grouh et al, 2011; Todorova et al, 2004; Zhang & Lespinasse, 1991; Zottini et al, 1997). Compared to the extensively performed rather low dose research on human and animal cells, the artificially applied radiation doses on pollen were very high and the results clearly demonstrate that pollen performance withstands much higher radiation levels.…”

Development and optimization of a germination assay and long-term storage forCannabis sativapollen

“…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, which requires a quick and effective method of determining pollen viability (Zottini et al, 1997;Engelmann and Takagi, 2000;Choudhary et al, 2014). Fluorescent stains such as fluorescein diacetate (FDA) or fluorochromatic reaction test (FCR) have been previously reported for assessing pollen viability in cannabis (Zottini et al, 1997;Choudhary et al, 2014). Viability is not always correlated with germination, as pollen may retain the ability to metabolize, while losing it's ability to germinate (Engelmann and Takagi, 2000).…”