Cannabis Seed Storage

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Product Description​

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https://www.amazon.com/AJFHKJ-Prote...in+fridge+holder+storage+case,aps,530&sr=8-1#

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Product details​

• Product Dimensions ‏ : ‎ 6.1 x 4.33 x 3.15 inches; 5.29 ounces

 

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untested ... in review
anyone have a preferred method ?

 

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In earlier research, induction of hermaphroditism in marijuana plants was achieved experimentally by applications of gibberellic acid (Heslop-Harrison, 1956, 1957; Ram and Jaiswal, 1970, 1972, 1974; Galoch, 1978; Rosenthal, 1991; United Nations Office on Drugs and Crime [UNOCD], 2009). Other studies showed that male and female flower ratios in marijuana plants could be altered by applications of chemicals such as 2-chloroethanephosphonic acid, aminoethoxyvinylglycine, silver nitrate, silver thiosulfate, or cobalt chloride (Ram and Jaiswal, 1970, 1972; Ram and Sett, 1981). Silver nitrate inhibits ethylene action in plants (Kumar et al., 2009) and was reported to increase male sex expression in marijuana, cucumber and gourd plants (Atsmon and Tabbak, 1979; Ram and Sett, 1982; Stankovic and Prodanovic, 2002). In a recent study, applications of silver thiosulfate induced male flower formation on genetically female hemp plants (Lubell and Brand, 2018). These findings demonstrate that changes in growth regulator levels in treated plants can impact hermaphroditic flower formation.

Physical or chemical stresses can also have a role in inducing staminate flower development on female plants of marijuana. For example, external environmental stresses, e.g., low photoperiods and reduced temperatures in outdoor production, were reported to increase staminate flower formation (Kaushal, 2012). Some plants formed hermaphroditic flowers when female plants were exposed to extended periods of darkness early during growth or during altered photoperiods during the flowering stage, although the exact conditions were not described (Rosenthal, 1991, 2000). Such stress factors could affect internal phytohormone levels, such as auxin:gibberellin ratios (Tanimoto, 2005), which could in turn trigger hermaphroditic flower formation in marijuana plants. In Arabidopsis plants, auxin, gibberellin and ethylene interact with jasmonic acid (JA) to alter stamen production (Song et al., 2013, 2014). Consequently, jasmonic-acid deficient mutant Arabidopsis plants exhibited male sterility, with arrested stamen development and non-viable pollen (Jewell and Browse, 2016) while JA treatment restored stamen development in these mutants. In marijuana plants, environmental stress factors which enhance JA production could potentially promote hermaphroditic flower formation but this requires further study. Lability of sex expression may offer advantages in promoting seed formation in hermaphroditic plants subject to environmentally stressful conditions (Ainsworth, 2000).

In the present study, pollen germination and germ tube growth were observed in samples of hermaphrodite flowers and pollen transfer from male flowers to stigmas of female flowers showed germination in situ followed by germ tube growth and penetration of the stigmatic papilla. Small and Naraine (2015) and Small (2017) showed pollen grains attached to stigmatic papillae but the germination and penetration process was not described. We observed a row of bulbous trichomes forming along the stomium on the anthers in staminate flowers and in hermaphroditic flowers, confirming earlier descriptions by Potter (2009) and Small (2017) for staminate flowers. The function of these trichomes is unknown. The findings described here are the first to demonstrate viable pollen production and anther morphology in hermaphroditic flowers in marijuana.

 

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Discussion​

Overall, STS at 3 mm was the most effective treatment for producing the greatest number of male flowers on female hemp plants. Green (2015) suggested that female hemp plants can be masculinized using a single foliar spray of 0.3 mm STS, but did not provide any information about the percent conversion to male flowers. We did not find three foliar sprays of 0.3 mm STS to be as effective for producing male flower formation as three foliar sprays of 3 mm STS. However, there may be specific strains, such as CBD hemp A, which produce primarily male flowers with STS concentrations less than 3mm.

Silver thiosulfate has been used to extend the vase life of cut flowers by blocking the effect of ethylene (Farnham et al., 1981; Veen and van de Geijn, 1978). A similar action of ethylene blocking by STS is likely responsible for the production of male flowers on female hemp plants in our study. It is generally believed that ethylene blocking is extended when a series of sprays of STS is used compared with a single spray of STS (Reid et al., 1980). It may be possible to induce male flowers on genetically female hemp using other ethylene perception inhibiting chemicals such as 1-methylcyclopropene.

Mohan Ram and Sett (1982), using 25 to 100 µg STS applied directly to the growing shoot tip of female hemp plants, were able to produce male flowers. However, they also noted severe necrosis on young leaves covering shoot tips and suspended apical growth for 20 to 25 d before lateral budbreak and subsequent flower formation. In comparison, we did not observe any plant phytotoxicity or delay in flower development. Furthermore, we were able to achieve 95% to 100% conversion to male flowers for all hemp strains, whereas Mohan Ram and Sett (1982) reported ≈60% to 80% conversion.

Producers and breeders should be able to masculinize female hemp plants routinely by using short-day conditions of ≈8 h and three foliar sprays of 3 mm STS at weekly intervals. We suspect that this method will be applicable for a broad range of genetically diverse hemp genotypes. Pollen produced by male flowers on genetically female plants can be used to produce all-female seed, but growers and breeders should be aware that pollen output may be reduced compared with pollen output from genetically male plants.

Foliar Sprays of Silver Thiosulfate Produce Male Flowers on Female Hemp Plants​

Article Category: Research ArticleOnline Publication Date: Dec 2018
Page(s): 743–747Volume/Issue: Volume 28: Issue 6
DOI: https://doi.org/10.21273/HORTTECH04188-18

 

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RAM, HY. "Mohan; SETT, R. Induction of fertile male flowers in genetically female Cannabis sativa plants by silver nitrate and silver thiosulphate anionic complex."
Theoretical and Applied Genetics 62.4 (1982): 369-375.

• H. Y. Mohan Ram &
• R. Sett

Summary​

Apical application of silver nitrate (AgNO3; 50 and 100 μg per plant) and silver thiosulphate anionic complex (Ag(S2O3)3−2; STS; 25, 50 and 100 μg per plant) to female plants of Cannabis sativa induced the formation of reduced male, intersexual and fully altered male flowers on the newly formed primary lateral branches (PLBs); 10 μg per plant of AgNO3 was ineffective and 150 μg treatment proved inhibitory. A maximum number of fully altered male flowers were formed in response to 100 μg STS. The induced male flowers produced pollen grains that germinated on stigmas and effected seed set. Silver ion applied as STS was more effective than AgNO3 in inducing flowers of altered sex. The induction of male flowers on female plants demonstrated in this work is useful for producing seeds that give rise to only female plants.

 

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From: Optimizing cannabis cultivation: an efficient in vitro system for flowering induction

Cannabis fertilization in vitro. a Male cannabis explant (cultivar Bt) flowering in vitro. b Cannabis male flower developed in vitro. c Pollen extracted from an in vitro anther. d In vitro germination of the extracted pollen. e In vitro inflorescence with developing seeds. In vitro-produced pollen was used to fertilize in vitro female plants (Sky 1 cultivar). f In vitro cannabis seed three weeks post-pollination g Germination of the hybrid seed in vitro. h The in vitro hybrid cannabis plant. Images b-g were captured using a stereomicroscope. Bar: b = 1000µm, c,d = 10µm, e,f,g = 100µm

 

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Tissue culture experimental system for flower induction in cannabis. a Stem segment explants, cut from indoor TA5 plants growing under 18/6 h light/dark photoperiod, were introduced into tissue culture (a white circle marks a node segment). b Stem section explants were surface sterilized in a 2% sodium hypochlorite solution. c The stems were separated into single-node segments, and five explants were cultured in each vessel. d Developed plants at two weeks under an 18/6 h light/dark photoperiod. e Cannabis flowers developed under a 12/12 h cycle to promote flowering. Images were taken three weeks into the flowering photoperiod. f A close-up of an in vitro cannabis flower (bar = 100µm). g and h Comparison between two photoperiod regimes. Node segments were introduced to tissue culture on the same date. g Vessels were immediately cultured under a flower-inductive photoperiod (12/12 h). h Vessels were cultured for two weeks under an 18/6 h cycle before switching to 12/12 h. Images were taken two weeks after introducing to tissue culture. i Average number of flowers per plant under two photoperiod regimes, with counts taken at specified times after introducing to tissue culture. The average flower number was calculated for five plants per vessel, presented as mean ± SE per plant for eight vessels for each treatment (n = 8); different letters represent a significant difference at a p < 0.05 using the Student's t-test
https://plantmethods.biomedcentral.com/articles/10.1186/s13007-024-01265-5#Fig1

Results​

We show that the life cycle of cannabis can be fully completed in tissue culture; plantlets readily produce inflorescences and viable seeds in vitro. Our findings highlight the superior performance of DKW medium with 2% sucrose in a filtered vessel and emphasize the need for low light intensity during flower induction to optimize production. The improved performance in filtered vessels suggests that plants conduct photosynthesis in vitro, highlighting the need for future investigations into the effects of forced ventilation to refine this system. All tested lines readily developed inflorescences upon induction, with a 100% occurrence rate, including male flowering. We revealed the non-dehiscent trait of in vitro anthers, which is advantageous as it allows for multiple crosses to be conducted in vitro without concerns about cross-contamination.

DKW basal medium supplemented with 2.32 μM KIN and 2.22 μM BA;​

 

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Flowering of cannabis under various media treatments. TA5 cannabis explants were cultured under an 18/6 h light/dark cycle for two weeks and then transferred to a 12/12 h flowering photoperiod. a Evaluation of the effect of activated charcoal in DKW medium compared to MS medium. The addition of activated charcoal significantly decreased flower production in the DKW medium and did not affect MS. Average flower numbers, presented as mean ± SE per plant, were calculated for five plants per vessel across four vessels for each treatment (n = 4). Different letters represent significant differences at p < 0.05 using the Tukey HSD test. Asterisks indicate significant differences between DKW and MS at p < 0.05 using the Student's t-test. b Impact of sucrose concentration in MS medium on flowering. Explants were cultured in MS medium, supplemented with four different concentrations of sucrose, across both vegetative and flowering photoperiods. Average flower numbers, presented as mean ± SE, were calculated for five plants per vessel across five vessels for each treatment (n = 5). Different letters represent significant differences at p < 0.05 using the Tukey HSD test. c The effect of sucrose concentration in DKW media on flowering. Average flower numbers, presented as mean ± SE, were calculated for five plants per vessel across three vessels for each treatment (n = 3); at p-value < 0.05 using Student t-test. d The effect of 6-Benzylaminopurine (6-BA) on cannabis flowering. 6-BA was added either during the vegetative phase only or during the flowering regime, as indicated in the index. Average flower numbers, presented as mean ± SE, were calculated for five plants per vessel across four vessels for each treatment (n = 4). Different letters represent significant differences at p < 0.05 using the Tukey HSD test

Stomatal abundance and chlorophyll content in cannabis leaves in tissue culture. a-c Comparison of TA5 cannabis stomata on the abaxial side of the fan leaves from tissue culture and growth room. a Scanning electron microscopy (SEM) image shows a similar stomata structure between the tissue culture and growth room. Bar = 50 µm. b, c. Stomata counting on abaxial fan leaves from tissue culture and growth room, using a light microscope (see Materials and Methods; bar = 20 µm). The number of stomata measured in 0.15 mm2 is presented as mean ± SE (n = 10), and different letters indicate significant differences at p < 0.05, using the Student's t-test. d-f Cannabis growth and chlorophyll content analysis in tissue culture without supplemented sugar. TA5 cannabis plants were cultivated in tissue culture either in a box (d) or a filtered box (e), with or without added sucrose. f Average chlorophyll content (measured in mg/g fresh weight) was calculated for two fresh leaves (number 3 and 4 from the apex) from each plant (a total of eight leaves per vessel). Leaves were weighed, and chlorophyll was extracted. Chlorophyll content was calculated according to spectrophotometric measurements at 645 and 663 nm (see Materials and Methods). Data are presented as mean ± SE for six vessels with four plants per vessel for each treatment (n = 6), and different letters represent a significant difference at p < 0.05 using the Tukey HSD test

 

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Flower Power: Floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa​

View ORCID ProfileA.S. Monthony, S. Bagheri, Y. Zheng, View ORCID ProfileA.M.P. Jones
doi: https://doi.org/10.1101/2020.10.30.360982
Now published in In Vitro Cellular & Developmental Biology - Plant doi: 10.1007/s11627-021-10181-5

 

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Flower power: floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa. | In Vitro Cellular & Developmental Biology - Plant​

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Fig. 3
The proposed micropropagation cycle using floral reversion. 1. Flowering: Culture of mature vegetative explants under a 12-hour photoperiod to trigger in vitro flowering. 2. Dissection: Single or pairs of florets are dissected. 3. Induction: Florets are transferred to DKW-based media to begin floral reversion. 4. Reversion: Normal vegetative growth occurs, indicating reversion. 5. Maturation: Reverted explants cultured on DKW will root and can be used again for in vitro flowering or moved ex vitro. 6. Acclimatization: Reverted explants can be transferred to ex vitro conditions for hardening.

Now published in In Vitro Cellular & Developmental Biology - Plant doi: 10.1007/s11627-021-10181-5

Flower Power: Floral reversion as a viable alternative to nodal micropropagation in Cannabis sativa

The legalization of Cannabis sativa L. for recreational and medical purposes has been gaining global momentum, leading to a rise in interest in Cannabis tissue culture as growers look for large-scale solutions to germplasm storage and clean plant propagation. Mother plants used in commercial...

www.biorxiv.org

Results​

Floral reversion​

Floret number significantly affected the percent reversion of explants to the vegetative state in both the BAP and mT experiments (Table 1 and Table 2). In both experiments, pairs were approximately three times more likely to revert than single florets. The treatment average for all BAP treated explants (0 μM to 10 μM) found that 55% of floret pairs reverted compared to only 18% of single floret (p < 0.0001). The BAP treatment with the highest percentage reversion was 1 μM BAP using pairs of florets which achieved an average of 69% reversion (Fig. 2A). A similar trend was observed in mT treated of floral explants of cv. U91, where pairs of florets showed approximately 2.5 times higher rate of reversion (70% vs 28%; p < 0.0001) between the singles and pairs of floret treatment averages. Treatment of pairs of florets at 1 and 10 μM mT achieved the highest percentage reversion with 81% of florets reverting (Fig. 2B). While the percent reversion was significantly affected by the floret number, the number of shoots produced per explant was not significantly affected by any of the fixed effects, with each treatment producing between 1.5 and < 2 vegetative shoots per responding explant. Each flowering in vitro plant produced an average of 24 ± 6 healthy florets (or 12 pairs) for use in the experiments.

 

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temp fluctuations in a standard food refrigerator
interesting for seed and maybe extended cold clone storage ?
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Refrigerators can help you meet regulations and guidelines from the CDC on vaccine storage and handling, USP<797> and USP <800> for clean rooms, and USP<1079> on managing the cold chain from manufacturing to cold storage. They are ideal for the storage of medications, vaccines and pharmaceuticals.

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US Pharmacopeia (USP)​

STATEMENTS/LABELING OF THE IMMEDIATE CONTAINERS OR PACKAGE INSERT




Storage statements

Cool Storage Statement—
The storage statement for labeling may be as follows: “Store in a cool place, 8 C to 15 C (46 Fto 59 F).”

Refrigerator Storage Statement—
The storage statement for labeling may be as follows: “Store in a refrigerator, 2 C to 8 C(36 F to 46 F).”

Freezer Storage Statement—
The storage statement for labeling may be as follows: “Store in a freezer, –25 C to –10 C (–13 F to 14 F).”

General Chapters_ _1079_ GOOD STORAGE AND SHIPPING PRACTICES.pdf

drive.google.com

Always wonder how purchased seed was handled prior to arrival was it frozen, how old is it, whats the germ%, if it was frozen previously should it be re-frozen ?

Best practice is to reproduce it and store it with germ% testing in the fridge or freezer ?
Seen some really great long term cold storage 10 years+

More testing to come