:D Genetic Preservation :D - Breeding

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• Research
• Open Access
• Published: 27 July 2022

Whole-genome resequencing of wild and cultivated cannabis reveals the genetic structure and adaptive selection of important traits​

BMC Plant Biology volume 22, Article number: 371 (2022) Cite

Background​

Cannabis is an important industrial crop species whose fibre, seeds, flowers and leaves are widely used by humans. The study of cannabinoids extracted from plants has been popular research topic in recent years. China is one of the origins of cannabis and one of the few countries with wild cannabis plants. However, the genetic structure of Chinese cannabis and the degree of adaptive selection remain unclear.

Results​

The main morphological characteristics of wild cannabis in China were assessed. Based on whole-genome resequencing SNPs, Chinese cannabis could be divided into five groups in terms of geographical source and ecotype: wild accessions growing in the northwestern region; wild accessions growing in the northeastern region; cultivated accessions grown for fibre in the northeastern region; cultivated accessions grown for seed in northwestern region, and cultivated accessions in southwestern region. We further identified genes related to flowering time, seed germination, seed size, embryogenesis, growth, and stress responses selected during the process of cannabis domestication. The expression of flowering-related genes under long-day (LD) and short-day (SD) conditions showed that Chinese cultivated cannabis is adapted to different photoperiods through the regulation of Flowering locus T-like (FT-like) expression.

Conclusion​

This study clarifies the genetic structure of Chinese cannabis and offers valuable genomic resources for cannabis breeding.

Except for seeds from Yunnan (W1) and Xizang (W2), the seeds from the other seven wild accessions all had a camouflage covering (a thin dark brown film attached to the surface of a seed), while only two accessions from Jilin (C7) and Anhui (C8) had a small amount of camouflage covering (Fig. 2). Moreover, wild cannabis bloomed earlier than domesticated cannabis. Although the flowering time of W1 and W2 was approximately 55 days, the flowering time of other wild cannabis accessions was shorter than 35 days (Table S1). In addition, the values of the first branch height, petiole length, compound leaf width and leaflet width of wild cannabis were significantly lower than those of cultivated cannabis (Fig. S1). We also observed that, when planted at low latitudes (Kunming), cultivated cannabis (C1-C7) from relatively high latitudes exhibited early flowering, early maturity, a dwarf stature and almost no branches (Fig. S1). However, wild cannabis plants still produced a relatively large number of branches in Kunming.

Unfortunately not any photos available of c1
We also observed that, when planted at low latitudes (Kunming), cultivated cannabis (C1-C7) from relatively high latitudes exhibited early flowering, early maturity, a dwarf stature and almost no branches (Fig. S1

 

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• Published: 29 April 2015

Expansion of female sex organs in response to prolonged virginity in Cannabis sativa (marijuana)​

• Ernest Small &
• Steve G. U. Naraine

Genetic Resources and Crop Evolution volume 63, pages 339–348

Abstract​

Female flowers of Cannabis sativa in wild-growing populations and in hemp plantations are almost always well supplied with pollen. The style-stigma portion of the pistils of such plants was found to average only about 3 mm in length and to invariably be two-branched. By contrast, “buds” (congested female inflorescences), the standard form of marijuana now produced in the illicit and medicinal marijuana sectors, are protected against pollen. This report documents that in the absence of pollen, the style-stigma parts of virgin pistils expand notably, average over 8 mm in length, and tend to develop more than two branches and to increase in girth. From an evolutionary viewpoint, this expansion of pollen-receptive tissue is an apparent adaptation for increasing the probability of fertilizing the females when males are extremely scarce. From a practical viewpoint, the expanded presence of stigma tissues may be both advantageous and disadvantageous. The high-THC secretory gland heads of Cannabis tend to fall away from marijuana buds, significantly decreasing pharmacological potency, but many gland heads become stuck to the receptive papillae of the stigmas, reducing the loss. Although stigmas constitute a small proportion of marijuana, their distinctive chemistry could have health effects.

Size matters: evolution of large drug-secreting resin glands in elite pharmaceutical strains of Cannabis sativa (marijuana)​

• Ernest Small &
• Steve G. U. Naraine

Genetic Resources and Crop Evolution volume 63, pages 349–359

Abstract​

Most tetrahydrocannabinol (THC) of Cannabis sativa is located in the resin heads of capitate-stalked glandular trichomes. We found that after harvest the resin heads shrink in diameter in exponential decay fashion under ambient room conditions, losing about 15 % in the first month, rising to 24 % over the first year, 32 % by 50 years, and 34 % after a century. An equation accounting for the asymptotic curve descriptive of the progression of shrinkage was determined [original gland head diameter in microns = observed diameter divided by (0.5255 + 0.4745 multiplied by time in days to the power −0.1185)], so that if the age of a specimen is known, the original diameter of the gland heads in the fresh state can be extrapolated. This equation was employed to compare gland head size in samples of different ages. A sample of high-THC medical marijuana strains marketed under license possessed resin head diameters averaging 129 μm, while a sample of low-THC industrial hemp cultivars possessed gland head diameters averaging 80 μm. The mean volume of the resin heads of the narcotic strains was more than four times larger than that of the industrial hemp strains. This is the first documented report of a consistent morphological separator of elite narcotic strains and non-narcotic plants. Most recognized strains of marijuana were bred clandestinely and illicitly during the last half century. The occurrence of large resin gland heads in a sample of officially marketed pharmaceutical strains is an obvious correlate of selection for higher quantity of resin production.

 

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What is a 1:1 Mating?

Single parent matings ie; one male one female. Excepting selfed plants, all matings have 2 parents. In a 1:1 mating you only use one male, preferably progeny tested. I prefer the seeds from the best progeny tested male rather than seeds that come from a mixed batch of males prior to testing. Only one can be the best.
N.

 

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Clarke & Merlin

X. Seed banks
When a Cannabis landrace is not reproduced every five
to ten years, the stored seeds will most likely die and the
landrace may be gone forever. Seeds must be properly
kept in a gene bank and reproduced periodically under
ideal conditions. The past 50 years have seen the genetic
diversity of the Cannabis genome dwindle away. Indeed,
the vast majority of landraces may already be extinct,
and we therefore must be careful to preserve and multi-
ply what remains. As Watson and Clarke (1997) warned,
“Many local landrace varieties, the result of hundreds of
years of selection for local use, have been lost because of
Cannabis suppression and eradication, neglect on the part
of agricultural officials and industry, anti-hemp propaganda
and the general trend (until recently) to reduce industrial
hemp breeding and research. Genetic materials are a living
heritage and we are their custodians. We must concentrate
our efforts to collect, preserve, characterize and utilize the
remaining Cannabis genetic resources before it is too late.”
As the worldwide reduction in Cannabis diversity
continues, the importance of genetic preservation
becomes more obvious. Unfortunately, no comprehen-
sive Cannabis germplasm collections exist. Most of the
few seed accessions are held by national gene banks that
may or may not share their valuable inventories with
breeders in other countries. The largest collection of
hemp germplasm is maintained by the Vavilov Institute
of Plant Research (VIR) in St. Petersburg, Russia. It pres-
ently numbers 563 seed accessions, including 23 possible
drug accessions from Afghanistan, Kazakhstan, Syria,
Turkey, and Uzbekistan, while the remaining are all
hemp and feral accessions from Armenia, Bulgaria,
Chile, China, Czechoslovakia, Estonia, France, Germany,
Hungary, Italy, Latvia, Moldova, Poland, Portugal,
Romania, Russia, Spain, Sweden, Ukraine, the United
States, and former Yugoslavia (Grigorev, 2015). Since the
late 1980s, political, technical, and financial difficulties in
Russia have resulted in low population sizes and incom-
plete isolation, and consequently there has been consid-
erable loss of genetic diversity and purity in the VIR
collection (Hillig, 2004b). Many accessions may now be
so similar to each other that their importance to future
breeding programs could be diminished.
In 1992, the Cannabis germplasm collection at Wage-
ningen University in the Netherlands contained over 156
accessions originating from 22 countries and largely
sourced from other collections and research institutes
(De Meijer and van Soest, 1992; Gilmore et al., 2007).
Nearly half of these accessions are from the former
USSR and Hungary. The Institute of Natural Fibres and
Medicinal Plants gene bank collections in Poland contain
139 accessions of predominantly European origin, with
accessions from France, Hungary, and the Ukraine
contributing 54.7% of the collection (Mankowska and
Silska, 2015). The Yunnan Academy of Social Sciences
collection in Yunnan province, China holds approxi-
mately 350 accessions mostly of East Asian origin
(Salentijn et al., 2014) and the Ecofibre Global
Germplasm Collection in Australia contains additional
Eurasian accessions (Welling et al., 2015). However,
comprehensive accession data are sorely lacking in
several of these collections and this limits their value to
breeders (Welling et al. 2016).
In addition, the subterranean Svalbard Global Seed
Vault on the Norwegian island of Spitsbergen about
1300 km (810 miles) from the North Pole has a total of
43 Cannabis accessions that are duplicated in three other
seed banks. Five of these accessions, from North Korea,
Netherlands, Spain, Syria, and Turkey, may possibly be
Cannabis drug populations; 21 others are hemp acces-
sions from Argentina, Austria, China, Croatia, France,
Georgia, Germany, Italy, Poland, Romania, Slovakia,
Spain, and Sweden; and 16 accessions are of unknown
origin (http://www.nordgen.org/sgsv/). The world’s larg-
est seed repository is the Millennium Seed Bank housed
at the Wellcome Trust Millennium Building in West Sus-
sex, near London, which specializes in wild plants and
has only one Cannabis accession, which is from Slovakia
(http://apps.kew.org/seedlist/SeedlistServlet).
Given the importance of Cannabis as a traditional as
well as present-day crop plant, the biodiversity of this
genus (particularly among the drug cultivars) is sorely
under-represented in seed banks, especially in light of
recent research interest in medical Cannabis. If we take
into account this lack of diversity, in light of genetic
impurity and low seed numbers, there really is no reliable
reserve of Cannabis seeds. The primary goal of germ-
plasm preservation is the conservation of the entire
genome of each population. It is especially important in
open-pollinated, cross-breeding plants that the popula-
tion size is large enough to ensure that as many of the
alleles as possible within each gene pool are reproduced
in the seed. A minimum of 1000 plants for monoecious
accessions, and 2000 plants for dioecious accessions,
assures that 99% of the Cannabis alleles will be repro-
duced (Crossa et al., 1993). Unfortunately, the seed
reserves of many of the Cannabis seed bank accessions
consist of less than 1000 viable seeds (often only 500 or
less); therefore, genetic diversity is already limited by the
number of archived seeds.
318 R. C. CLARKE AND M. D. MERLIN

The secondary goal of genetic
preservation is to reproduce the accessions in sufficient
quantities to maintain a reserve for future reproductions
and public distribution.
A common goal of Cannabis breeders should be
establishing a more comprehensive core collection of
Cannabis seed accessions that have been exhaustively
characterized agronomically in the field, and on molecu-
lar levels, genetically and chemically, in the laboratory.
Only then, can we see what diversity really is available
for researchers to work with in the future. This core col-
lection should be maintained with optimal reproduction
and storage methodology, and individual accession eval-
uations should be made accessible to breeders (Watson
and Clarke, 1997).
In the past 20 years the situation has only stagnated.
According to Welling et al. (2016), in their fine review of
the present state of ex situ Cannabis germplasm
collections:
“Coordinated and comprehensive conservation and
characterization of ex situ Cannabis resources holds the
promise of preserving genepool diversity and enabling cul-
tivar development. However, the legal constraints imposed
by international narcotics conventions over more than
50 years have been influential in the fractionation and
erosion of publicly accessible Cannabis ex situ genetic
resources. The restrictions on legal exchange of bona fide
research materials continues to limit the establishment of
physical and centralized ex situ core collections.”

XI. Present and future directions for Cannabis
breeding start here!
The primary evolutionary process that is presently fur-
thering domestication in Cannabis is basic Mendelian
breeding; this traditional system involves selecting sim-
ply inherited traits and increasing their homozygosity
through sexual recombination. Many of the primary eco-
nomic traits of Cannabis are simply inherited (e.g., stalk
height, seed size, and cannabinoid content). Cannabi-
noid biosynthesis is controlled by a narrow range of
alleles limited to only a few loci; heritability is extremely
high, which has favored successful breeding for high-
THC and high-CBD sinsemilla cultivars as well as low-
THC industrial hemp cultivars. High variability and
strong heritability also influence both flowering and
maturity times, and breeding for earlier-maturing drug
varieties and later-maturing fiber varieties continues.


XI. Present and future directions for Cannabis breeding start here!
The primary evolutionary process that is presently fur-
thering domestication in Cannabis is basic Mendelian
breeding; this traditional system involves selecting sim-
ply inherited traits and increasing their homozygosity
through sexual recombination. Many of the primary eco-
nomic traits of Cannabis are simply inherited (e.g., stalk
height, seed size, and cannabinoid content). Cannabi-
noid biosynthesis is controlled by a narrow range of
alleles limited to only a few loci; heritability is extremely
high, which has favored successful breeding for high-
THC and high-CBD sinsemilla cultivars as well as low-
THC industrial hemp cultivars. High variability and
strong heritability also influence both flowering and
maturity times, and breeding for earlier-maturing drug
varieties and later-maturing fiber varieties continues.

 

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assam flower

Ganja (Assamese: গাঁজা)

Assam Hash Plant by:ILT

coco's assam seed for repro free share

Assam is a state in northeastern India known for its wildlife, archeological sites and tea plantations. In the west, Guwahati, Assam’s largest city, features silk bazaars and the hilltop Kamakhya Temple. Umananda Temple sits on Peacock Island in the Brahmaputra river. The state capital, Dispur, is a suburb of Guwahati. The ancient pilgrimage site of Hajo and Madan Kamdev, the ruins of a temple complex, lie nearby. ― Google

 

[Plookerkingjon]

Hopefully this thread is a seed that once planted begins to grow,
with community love and support and blessing may it grow into a large tree!
Hopefully its branches reach around the globe to every small village and tribe.

To promote biodiversity and preservation not greed but giving.
Protect the planet and people from corporate greed, may the corporations
always remember that there are people at the heart of every corporation.

More to the point
Genetic preservation its been said we need thousands of plants to maintain.
In this thread im going to suggest whats feasible for us to all do our part.

If you try and preserve a Cannabis landrace with less than 1,000 females from seed and 1,000 males from seed you will lose genes every reproduction that is not real preservation.
Some think the work can be staggered and only 100 reproduced each crop, for 10 consecutive years, but that is not how genetics work you need all 1,000 females and 1,000 males freely pollinating each other, reproduced at the same time in the same location to preserve a landrace and avoid gene loss. Cannabis is a Dioecious, Heterozygous, Obligate Outcrosser, that is why 1,000 males and 1,000 females from seed are required for preservation to keep all the genepool intact for a given landrace.-SamS
Statistical genetic considerations for maintaining germ plasm collections
J. Crossa , C. M. Hernandez , P. Bretting , S. A. Eberhart , S. Taba
Theor Appl Genet (1993) 86:673-678
DOI: 10.1007/BF00222655

Methodologies for estimating the sample size required for genetic conservation of outbreeding crops.
Crossa, J.
Theoretical and Applied Genetics, 77(2). (1989).
doi:10.1007/bf00266180

In my own situation for example im going to try to preserve a
"indica" and a "sativa" these terms used loosely
for this project im going to look a effect, leaf shape and latitude of origin


for the sativa im going to pick a sativa from a equatorial sativa (?)
maybe Kerala open to suggestions, please post below

for the indica im going to pick a indica type from the snow Himalayas (?)
maybe siberia im open to suggestions,
please post below maybe include a specimen photo

Additionally im looking to aquire 20-40 seeds for a strain
100-200 seeds for genetic preservation
(if a few of us keep a genetic preservation line we could as a collective hold 1000s of seeds)


Also this will aid in personal breeding projects specifically im looking for
plant structure
bud structure
calyx to leaf ratio
maturity time
effect / duration
terpene content
potency
vigor
unique mutations


if you have anything to ask add share here, only good vibes please !

let it grow!

I'm new to this bear with me, and from time to time when the arthritis acts up in my right hand I tend to use the Google mic so my vernacular may be ongoing rambling sentences or paragraphs it's not done intentionally yes I'm a lazy bastard when I'm in full arthritic whatever you call it. So when they say a thousand females in a thousand males are they talking about letting them openly pollinate or could those hectares be broken up bearing in mind that I was just told this past summer that cannabis Paul and can travel up to 5 miles I think I may be wrong I have a lot to learn.
I would also like to state that this one post when I pictured in my imagined my mind my imagination runs wild and I think to myself for kids that grew up born in the late 60s early 70s what this describes is a property that is a bucket list to work on for a person like me

 

[acespicoli]

I'm new to this bear with me, and from time to time when the arthritis acts up in my right hand I tend to use the Google mic so my vernacular may be ongoing rambling sentences or paragraphs it's not done intentionally yes I'm a lazy bastard when I'm in full arthritic whatever you call it. So when they say a thousand females in a thousand males are they talking about letting them openly pollinate or could those hectares be broken up bearing in mind that I was just told this past summer that cannabis Paul and can travel up to 5 miles I think I may be wrong I have a lot to learn.
I would also like to state that this one post when I pictured in my imagined my mind my imagination runs wild and I think to myself for kids that grew up born in the late 60s early 70s what this describes is a property that is a bucket list to work on for a person like me

1-2000 plants needed to preserve 99% of a landrace strain
Not that those numbers should limit anyone in doing their small part in saving even just one seed

There is a dream and then there is reality


XII. Summary and conclusions
Cannabis is an annual, sun-loving plant that thrives in open,
nitrogen-rich environments, including rubbish piles created
by humans (Anderson, 1967; Merlin, 1972; Clarke and
Merlin, 2013; also see Small, 2015). Cannabis also has rela-
tively large, numerous, and easily sown seeds, and therefore
was preadapted for cultivation. Close associations between
humans and Cannabis stimulated its early cultivation, and
over time this eventually led to its domestication. As ancient
agricultural strategies took form, humans began to select
plants that provided more and better products. Some of the
earliest seed selections perished and growers must have
constantly collected from spontaneous populations to
replace, supplement, or modify extant varieties, resulting in
evolution of improved landraces. As their familiarity with
Cannabis grew, humans became more discerning with their
seed selections.
Because Cannabis grew well as a camp follower, later
selections were increasingly made from feral populations
near human settlements, rather than truly wild popula-
tions. Early semi-domesticated varieties also became nat-
uralized and interbred with nearby feral and/or wild
populations. Selection for differing economic traits con-
tinued as Cannabis spread well beyond its original puta-
tive range in Central Asia. Backcrosses between
cultivated and truly wild populations became rare,
whereas crosses between cultivated and feral populations
became more common. Eventually, isolation developed
between wild, cultivated, and feral populations which
evolved their own phenotypes. This scenario led to the
extreme variation encountered today in geographically
isolated populations,

 

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Master Kush is an indica/sativa variety from Nirvana and can be cultivated indoors (where the plants will need a flowering time of ±70 days) and outdoors. Nirvanas Master Kush is a THC dominant variety and is/was also available as feminized seeds.

ManicMiner

Nirvanas Master Kush Description​

First called High Rise, Master Kush was developed in one of the tall buildings in Amsterdam's Bijlmer area. Coffeeshop owners and regulars alike fell head over heels in love with this exclusive tetraploid strain. By popular demand, this Hindu Kush / Skunk hybrid was stabilized and marketed, and it has been a classic ever since. A strong plant of medium height and bushiness, Master Kush is a heavy producer which thrives in soil, hydro and greenhouse growing systems. Master Kush has a pleasantly earthy, mossy smell and its smoke is smooth.
Yield: 400 - 500 grams/m² (SoG)
Effect: High and Stoney
Grow height: Medium
Flowering Indoor: 9 / 11 weeks

Frontiers | Polyploidization for the Genetic Improvement of Cannabis sativa

Cannabis sativa L. is a diploid species, cultivated throughout the ages as a source of fiber, food, and secondary metabolites with therapeutic and recreation...

www.frontiersin.org

​

Although this method has been used in hemp-type Cannabis, it has never been applied to drug-type strains. Here, we describe the development of tetraploid drug-type Cannabis lines and test whether this transformation alters yield or the profile of important secondary metabolites: Δ9-tetrahydrocannabinol (THC), cannabidiol (CBD), or terpenes. The mitotic spindle inhibitor oryzalin was used to induce polyploids in a THC/CBD balanced drug-type strain of Cannabis sativa. Cultured axillary bud explants were exposed to a range of oryzalin concentrations for 24 h. Flow cytometry was used to assess the ploidy of regenerated shoots. Treatment with 20–40 μM oryzalin produced the highest number of tetraploids. Tetraploid clones were assessed for changes in morphology and chemical profile compared to diploid control plants. Tetraploid fan leaves were larger, with stomata about 30% larger and about half as dense compared to diploids. Trichome density was increased by about 40% on tetraploid sugar leaves, coupled with significant changes in the terpene profile and a 9% increase in CBD that was significant in buds. No significant increase in yield of dried bud or THC content was observed. This research lays important groundwork for the breeding and development of new Cannabis strains with diverse chemical profiles, of benefit to medical and recreational users.

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Master Kush, also known as "High Rise," "Grandmaster Kush," and "Purple SoCal Master Kush" is a popular indica marijuana strain crossed from two landrace strains from different parts of the Hindu Kush region by the Dutch White Label Seed Company in Amsterdam.

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el natural is my first choice, interesting no less. Nice to know your smoke is organic?
one fellow related as a comparison to a seeded vs non seeded watermelon, study that if you have not

 

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Production of Tetraploid and Triploid Hemp​

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https://journals.ashs.org/hortsci/view/journals/hortsci/55/10/article-p1703.xml DOI: https://doi.org/10.21273/HORTSCI15303-20 Page Count: 5 Volume/Issue: Volume 55: Issue 10​

Article Category: Research Article Online Publication Date: 18 Sep 2020

Abstract​

To maximize yield, cannabidiol (CBD) hemp producers prefer female plants, and this is accomplished by using expensive feminized seed, vegetatively propagated female clones, or by removing male plants from dioecious seed lots. Hemp pollen drifts long distances on wind, and pollination of females reduces CBD content. Induction of triploidy is a common strategy used by plant breeders to produce sterile cultivars of agricultural crops. Triploid (3n) hemp, with three sets of chromosomes, was developed by crossing naturally diploid (2n) hemp with tetraploid (4n) hemp. Tetraploid plants used to create triploids were produced using pregerminated seeds and the mitotic spindle inhibitor colchicine. Seedlings from seeds of ‘Abacas’ × [(‘Otto2’ × ‘BaOx’) × (‘BaOx’ × ‘Colorado Cherry’)] treated with 0.05% colchicine or 0.02% colchicine for 12 hours and longer were significantly shorter than controls and ≤1 cm tall at 10 days after sowing. Surviving seedlings exhibited thickened cotyledons and hypocotyls, which indicated a potential change in ploidy. Tetraploid induction ranged from 26% to 64% for pregerminated seeds of five different hemp cultivars (Abacus × Wife, Cherry Wine, Mountain Mango, Wife, and Youngsim10) treated with 0.05% colchicine for 12 hours. Tetraploids had nearly twice the DNA content as diploids according to flow cytometric analysis. Tetraploid ‘Wife’ had larger stomates and reduced stomatal density compared with diploid ‘Wife’. Four triploid ‘Wife’ genotypes produced from crossing tetraploid ‘Wife’ with diploid ‘Wife’ were acclimated to greenhouse conditions after embryo rescue. DNA content and stomate size of triploid ‘Wife’ was intermediate between the parents. This is the first report of triploid plants of hemp. Future research will evaluate the sterility of triploid hemp.
Keywords: Cannabis sativa; colchicine; embryo rescue; flow cytometry; pregerminated seed
Cannabis sativa (hemp, marijuana) is a dioecious species with homogametic (XX) pistillate female plants and heterogametic (XY) staminate male plants (Moliterni et al., 2004). The species is cultivated for cannabinoids, most notably CBD and tetrahydrocannabinol (THC), fiber, and grain, from which a wide range of consumer products are derived (Small, 2015). Cannabinoids have reported medicinal value and are produced in the glandular trichomes of the plant, which are found in the greatest density on the inflorescences of female plants (Small and Cronquist, 1976). Hemp is distinguished from marijuana by the content of THC produced by the plant, which is less than 0.3% dry weight THC for hemp (Agriculture Marketing Service, 2019). Hemp fiber is produced from the stalks and the seed is harvested for grain and hempseed oil (Small, 2015). Monecious cultivars have been developed for dual-purpose fiber and grain production.
Hemp seed from open-pollinated dioecious plants can be expected to produce a 50:50 ratio of male-to-female plants (Small, 2015). During CBD hemp production, it is important for growers to remove male plants before anthesis, because pollination of female plants reduces cannabinoid yield (Meier and Mediavilla, 1998). Hemp growers prefer to use feminized seed or vegetatively propagated female clones for CBD production to eliminate the labor of removing male plants and the lost acreage from removed male plants. Hemp is wind-pollinated, and pollen can drift long distances (Small, 2015). It has been reported that hemp pollen can drift more than 300 km (Clarke, 1977). Therefore, even when hemp farmers take strict measures to grow only female plants, they can experience seed production as a result of drifting pollen from neighboring fiber and grain farms or from CBD farms that did not remove males. A distance of at least 5 km is recommended to prevent pollen drift from neighboring hemp fields (Neiden, 2020; Small, 2015). Disputes between farmers over unintended seed production from drifting pollen has led to several lawsuits (Perkowski, 2019). Pollen can also drift from wild or escaped hemp, known as ditch weed (Neiden, 2020).
Induction of polyploidy has been used by plant breeders to develop improved horticultural crops with enhanced traits such as size, vigor, and metabolite content (Alexander, 2017; Lehrer et al., 2008; Sattler et al., 2016; Wang et al., 2016; Xu et al., 2014). Tetraploids are polyploids that contain four sets of chromosomes. Compared with diploids, tetraploid purple cone flower produces more secondary metabolites and biomass (Xu et al., 2014), and tetraploid ryegrass is more drought tolerant and disease resistant (Sattler et al., 2016). Tetraploidy can be induced artificially using mitotic spindle inhibitors such as colchicine or oryzalin (Sattler et al., 2016; Wang et al., 2016). C. sativa is almost exclusively diploid (2n = 20) in the wild (Small and Cronquist, 1976). There is only one report of a natural tetraploid of C. sativa, from India (Sharma et al., 2015). Tetraploid C. sativa has been produced using colchicine on seedling shoot tips (Bagheri and Mansouri, 2015; Mansouri and Bagheri, 2017), and by using oryzalin on in vitro nodal explants (Parsons et al., 2019). Tetraploid plants produced in these studies exhibited traits such as larger leaves and greater shoot fresh weight and flavonoid content.
Tetraploid plants crossed with diploid plants can generate triploid plants, which have three sets of chromosomes (Wang et al., 2016). Triploid plants are frequently seedless, because unequal segregation of chromosome pairs during meiosis results in inviable gametes (Wang et al., 2016). Seedless triploid cultivars have been bred for hops, watermelon, banana, and citrus (Trojak-Goluch and Skomra, 2018; Wang et al., 2016). Warmke and Davidson (1944) reported crossing tetraploid and diploid marijuana and producing triploid plants; however, no cytogenetic evidence of triploidy was provided. The objective of this work was to investigate a more efficient and easy method for inducing tetraploidy in hemp, and to cross tetraploid plants with diploid plants to produce triploid hemp. Triploid hemp that does not produce seed when exposed to pollen could be a solution for the problem of pollen drift.

Materials and Methods​

​

Tetraploid development.​

Two experiments were conducted to produce tetraploid hemp plants by treating pregerminated seeds with colchicine. In Expt. 1, pregerminated seeds of ‘Abacas’ × [(‘Otto2’ × ‘BaOx’) × (‘BaOx’ × ‘Colorado Cherry’)] were exposed to two colchicine concentrations (0.02% or 0.05%) for three durations (6, 12, or 24 h) to determine a suitable exposure rate for tetraploid induction. Control pregerminated seeds were treated with water for 24 h. In Expt. 2, pregerminated seeds of five different hemp cultivars (Abacus × Wife, Cherry Wine, Mountain Mango, Wife, and Youngsim10) were treated with 0.05% colchicine for 12 h to generate additional tetraploid genotypes. Seeds were pregerminated by soaking them in water for 24 h and then transferring them to 100- × 15-mm petri dishes lined with moistened filter paper (Whatman no. 4; Whatman, Maidstone, UK) for another 24 h. After this treatment, seeds were considered pregerminated because radicals had emerged from 1 to 5 mm (Fig. 1A). For the colchicine treatment, pregerminated seeds were placed in 50-mL conical tubes that were agitated gently at a slow speed on a platform shaker for the duration of treatment.

Fig. 1.
Photographs of (A) pregerminated seeds with insert showing thickened hypocotyl of colchicine-treated seedling (B) germinated, and rescued triploid embryo (C) three triploid genotypes of parentage W1 × ‘Wife’ established in 1-L containers.
Citation: HortScience horts 55, 10; 10.21273/HORTSCI15303-20

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For Expt. 1, there were 25 pregerminated seeds per treatment, for a total of 175 seeds. For Expt. 2, the number of pregerminated seeds treated per cultivar varied because of limited seed availability. After colchicine treatment, seeds were rinsed with deionized water and sown to a depth of 6 to 8 mm in 50-cell plug trays using a peatmoss-based seed starting mix (Fafard 3B Mix; Sungro Horticulture, Agawam, MA). For Expt. 1, seedlings were grown in a randomized complete block design, with five seedlings per experimental unit and five blocks (n = 5). Plants from both experiments were grown in a greenhouse with set points of 21 °C/17 °C day/night under long-day conditions (18 h light). Plants were fertigated as needed with a soluble fertilizer (Peters 20N–8.7P–16.6K; Scotts, Marysville, OH), providing 100 ppm nitrogen (N). On day 10 after sowing, percent emergence and seedling height were recorded. Pregerminated seeds were considered emerged if cotyledons were visible above the soil surface. Percent emergence was calculated by dividing the number of seedlings emerged by the total number of seeds sown. For Expt. 1, percent emergence was calculated per experimental unit. Tetraploid induction rate was calculated by dividing the number of tetraploids produced by the total number of seeds treated, and then multiplying by 100%. Data were subjected to analysis of variance (ANOVA) (Proc GLM) using SAS (version 9.4; SAS Institute, Cary, NC). Mean separation using Tukey’s honestly significant difference test (P ≤ 0.05) was performed for percent emergence and seedling height.

Triploid development.​

On 5 Dec. 2019, three clonal plants each of two tetraploid, female ‘Wife’ genotypes (W1 and W2), established in 2-gallon containers filled with a peatmoss-based potting mix (Promix BK25; Premier Tech Horticulture, Quakertown, PA), were provided short days (12 h) to induce flowering in a greenhouse with set points of 21 °C/17 °C day/night. In addition, four diploid, female, clonal ‘Wife’ plants established in 1-gallon containers using the same potting mix were also provided short days in the same greenhouse. Plants were top-dressed with controlled-release fertilizer (Osmocote Plus 15N–3.9P–10K 5- to 6-month formulation; Everris NA, Dublin, OH) at 10 g per 1-gallon container and 30 g per 2-gallon container. Plants received a soluble fertilizer (Peters 15N–12.9P–12.5K, Scotts) providing 100 ppm N at every irrigation (1 L/d and 2 L/d for 1-gallon and 2-gallon container plants, respectively). On the first day of short days, diploid plants were sprayed with silver thiosulfate solution at a concentration of 3 mm prepared according to Lubell and Brand (2018) to induce male flower development. These plants were sprayed to runoff (≈200 mL/plant) on three occasions, 7 d apart, on 5 Dec., 12 Dec., and 19 Dec. 2019.
Anthesis of male flowers produced on diploid female ‘Wife’ plants began on 2 Jan. 2020. Hand-pollination of tetraploid ‘Wife’ plants with feminized (all female) diploid ‘Wife’ pollen occurred from 2 Jan. 2020 to 10 Jan. 2020. Tetraploid plants were used as the maternal parent, because the 2n female gamete has been shown to be more efficient than 2n pollen for the formation of triploid pants (Wang et al., 2016). Feminized pollen was collected in 50-mL conical tubes by gently agitating male flowers with a gloved finger to release pollen. Collected pollen was brushed on female tetraploid stigmas using a soft bristled paintbrush. Putative triploid seeds developed normally on tetraploid mothers for the first 2 weeks after pollination, at which time seed development appeared to arrest and seeds started to become soft. For this reason, on 24 Jan. 2020, embryo rescue was attempted for the remaining seeds that were firm. The majority of developing seeds failed, became soft, and were not useful. From tetraploid plants, 20 remaining seeds were harvested and surface-sterilized with 20% sodium hypochlorite solution, and the embryos were excised under sterile conditions. Fifteen embryos were excised successfully from the immature seeds and introduced into aseptic culture. Immature embryos were cultured in G7 magenta boxes containing 45 mL Murashige and Skoog medium supplemented with 3% sucrose, 2 µm meta-Topolin, and 0.8% agar at pH 5.7. Seven embryos germinated (Fig. 1B) and were subcultured to the same medium on 21 Feb. 2020. At subculture, roots were removed from the embryos and hypocotyls were trimmed to 1 cm below the cotyledons to enhance expansion of the epicotyl. On 20 Mar. 2020, putative triploid shoots were rooted ex vitro in rockwool cubes. Four rooted plants were potted in 1-L containers using the same potting medium stated previously and acclimated to the greenhouse with set points as described and long days.

Flow cytometry.​

Flow cytometric analysis was used to determine ploidy level as was done for C. sativa by Bagheri and Mansouri (2015) and Parsons et al. (2019). For flow cytometric analysis, 50 mg of young leaf tissue was harvested per plant. A modified version of the protocol noted in Arumuganathan and Earle (1991), and described in Lehrer et al. (2008) and Mahoney et al. (2019), was followed. Young leaves were chopped using a razor blade in nuclei suspending solution in a 55-mm petri dish on ice. The suspending solution was filtered, centrifuged to form a pellet, and resuspended in nuclei staining solution containing propidium iodide. Relative fluorescence of total DNA (FL2-A) was measured using a flow cytometer (BD Accuri C6; BD Biosciences, San Jose, CA). Data were displayed in histograms using BD Accuri C6 Software (1.0.264.21; BD Biosciences, San Jose, CA). To determine ploidy levels of putative tetraploids and triploids, their histogram peaks were compared with those derived from diploid hemp. Flow cytometric analysis was conducted five times over four clonal propagation cycles for three tetraploid ‘Wife’ genotypes to document ploidy stability.

Stomata characteristics.​

Stomata of diploid, triploid, and tetraploid genotypes of ‘Wife’ were evaluated. To visualize stomata, nail polish impressions of the abaxial surface of mature fan leaves were made for three leaves per genotype (Grant and Vatnick, 2004). Impressions were visualized using a compound microscope (Microphot-FXA; Nikon Instruments, Melville, NY) and microscopy camera (Infinity3; Teledyne Lumenera, Ottawa, ON). Length and width for 10 stomata per leaf were measured at 10× magnification using ImageJ software (version 1.52a; National Institutes of Health, Bethesda, MD). The number of stomata present in a 0.04-mm2 area of leaf was counted. Stomata density was calculated by dividing the number of stomata by 0.04.
Data were subjected to ANOVA (Proc GLM) using SAS (version 9.4; SAS Institute). Mean separation using Tukey’s honestly significant difference test (P ≤ 0.05) was performed for stomata density and stomata length and width.

Results and Discussion​

Colchicine-treated seedlings were significantly shorter than untreated control seedlings 10 d after sowing (Table 1). Seedlings treated with 0.02% colchicine for 24 h and 0.05% colchicine for all durations were less than 1 cm tall at 10 d after sowing and exhibited thickened cotyledons and hypocotyls (Fig. 1A, insert). Some of these seedlings failed to expand their epicotyl and perished. At 0.02% colchicine, percent emergence and seedling height decreased as duration of exposure to colchicine increased from 6 to 12 to 24 h. In total, five tetraploids were produced from Expt. 1. Three of the five tetraploids were generated from exposure to 0.02% colchicine for 12 h. This treatment also resulted in two mixoploids. The remaining two tetraploids were produced from exposure to 0.05% colchicine for 6 and 12 h. At 0.05% colchicine, the 6-h treatment produced three mixoploids, whereas the 12-h treatment produced no mixoploids.
Table 1.
Percent emergence, seedling height, and number of tetraploid and mixoploid plants produced per treatment from exposing pregerminated seeds of ‘Abacas’ × [(‘Otto2’ × ‘BaOx’) × (‘BaOx’ × ‘Colorado Cherry’)] to 0.02% or 0.05% colchicine for 6, 12, or 24 h.

For Expt. 2, our goal was to produce tetraploids of more desirable cultivars for which we had limited seed quantity. We decided to treat with 0.05% colchicine for 12 h to ensure production of only seedlings with short stature and thickened cotyledons and hypocotyl, and to minimize the number of mixoploids. Tetraploid induction ranged from 26% to 64%, depending on the cultivar in Expt. 2 (Table 2). A total of 39 tetraploids were generated in Expt. 2, which is notably more than those generated in Expt. 1, which may be attributed to enhanced seed quality or genotype response to colchicine in Expt. 2. More C. sativa tetraploids were produced using pregerminated seeds in Expt. 2 than were produced by Bagheri and Mansouri (2015) using 7-d-old seedlings and colchicine, and by Parsons et al. (2019) using nodal explants and oryzalin. We attempted tetraploid induction according to Bagheri and Mansouri (2015) using 0.2% colchicine, but were unsuccessful at generating polyploids (data not shown). Our findings demonstrate that using pregerminated seeds and colchicine is an easy and effective method for producing tetraploids of hemp. This method was used successfully by Lehrer et al. (2008) to produce tetraploids of Japanese barberry.

Table 2.
Percent emergence and number of tetraploid and mixoploid plants produced from treating pregerminated seeds of five hemp cultivars with 0.05% colchicine for 12 h.

Parsons et al. (2019) reported that the ploidy of induced tetraploids of C. sativa was stable through clonal propagation. We found for three induced tetraploid ‘Wife’ genotypes that ploidy was stable over four cycles of clonal propagation by stem cuttings.

Four triploid hemp genotypes were confirmed using flow cytometry and analysis of stomatal characteristics. Three genotypes had the parentage W1 × ‘Wife’ (Fig. 1C) and the fourth genotype had the parentage W2 × ‘Wife’. FL2-A fluorescence of total DNA was 225,000 for the tetraploids, which is a 96% increase in DNA content over the diploids with FL2-A fluorescence of 115,000 (Fig. 2). Similarly, Bagheri and Mansouri (2015) and Parsons et al. (2019) found that the DNA content of tetraploid C. sativa was almost twice that of diploid C. sativa according to flow cytometric analysis. The FL2-A fluorescence of the triploid was 160,000, which is 40% more DNA than the diploid and 40% less DNA than the tetraploid. The intermediate FL2-A finding for triploid hemp was precisely what we expected for plants with 3n chromosomes.

Fig. 2.
Flow cytometric histograms representing hemp plants with (A) a diploid (2n) profile, (B) a triploid (3n) profile, and (C) a tetraploid (4n) profile. FL2-A = relative fluorescence of total DNA.
Citation: HortScience horts 55, 10; 10.21273/HORTSCI15303-20

• Download Figure

Stomate length and width for ‘Wife’ genotypes increased significantly as ploidy increased from diploid to triploid to tetraploid (Table 3, Fig. 3). Stomatal density was greater for diploid and triploid ‘Wife’ than for tetraploid ‘Wife’. Parsons et al. (2019) found that tetraploid C. sativa had larger stomates and reduced stomatal density than diploid C. sativa. Similarly, for coffee, as ploidy level increased, stomate size increased and stomatal density decreased (Mishra, 1997). The larger cell sizes observed for tetraploid and triploid plants compared with diploids could be the result of increased copies of genes present for the polyploids (Sattler et al., 2016).

Table 3.
Stomata density, determined for an area of leaf measuring 0.04 mm2, and stomate length and width for diploid, triploid, and tetraploid plants of hemp cultivar Wife.

Fig. 3.
Nail polish impressions showing stomata on the abaxial surface of (A) diploid (B) triploid and (C) tetraploid fan leaves. Scale bars = 20 µm.
Citation: HortScience horts 55, 10; 10.21273/HORTSCI15303-20

• Download Figure

The method reported here of exposing pregerminated seed to a mitotic spindle inhibitor such as colchicine represents a relatively easy way to produce tetraploids of hemp. A possible drawback of this seed-based method is the genetic variability inherent in some seed. To our knowledge, this is the first report of triploid plants of hemp. Future research is needed to evaluate the sterility and growth performance of hemp triploids. Seedless triploids have been developed for hops, a close relative of C. sativa (Trojak-Goluch and Skomra, 2018). In addition to sterility, triploid hops have proved to be higher yielding and have increased alpha acids, which are important for aroma and flavor. It is possible that triploid hemp may offer some unique cannabinoid profiles or increased secondary metabolite content.

 

[acespicoli]

how things change, mailing clones circa 2022

No agar in it. You have to add that separately, if needed.
Designed for the growth of tobacco plants

Murashige and Skoog medium (or MSO or MS0 (MS-zero)) is a plant growth medium used in the laboratories for cultivation of plant cell culture. MSO was invented by plant scientists Toshio Murashige and Folke K. Skoog in 1962 during Murashige's search for a new plant growth regulator. A number behind the letters MS is used to indicate the sucrose concentration of the medium. For example, MS0 contains no sucrose and MS20 contains 20 g/L sucrose. Along with its modifications, it is the most commonly used medium in plant tissue culture experiments in the laboratory,[1] but according to recent scientific findings it should not be used as nutrient solution in deep water culture.[2]

As Skoog's doctoral student, Murashige originally set out to find an as-yet undiscovered growth hormone present in tobacco juice. No such component was discovered; instead, analysis of juiced tobacco and ashed tobacco revealed higher concentrations of specific minerals in plant tissues than were previously known. A series of experiments demonstrated that varying the levels of these nutrients enhanced growth substantially over existing formulations. It was determined that nitrogen in particular enhanced growth of tobacco in tissue culture.

with you carbon / exhaust fans and a small grow tent add a HEPA filter and you have a DIY quick dual purpose laminar flow air hood. Add some sterilization clean room and sterile technique

 

[acespicoli]

Wild cannabis is one of the many plant species making up the rank vegetation at the well-watered open woodland along the G216. (Craig Brelsford)

International Crisis

Xinjiang, Charas, and the Origins of Cannabis - The Real Seed Company

Xinjiang was once at the heart of global cannabis production, supplying Central Asian sieved charas (hashish) to what was the world’s largest market for cannabis drugs, India. The Xinjiang charas trade came to a sudden halt in the 1930s following...Read more

therealseedcompany.com

 

[romanoweed]

where did you find the Photo of the Plant growing along th G216 Road?

That Pheno looks relatively good.

 

[acespicoli]

where did you find the Photo of the Plant growing along th G216 Road?

That Pheno looks relatively good.

Craig Brelsford took the photo he is a bird watcher with a huge camera that traveled thru china look him up on his blog he may be able to answer more questions about hemp bird seed food, where it grows what birds eat it etc...are the birds effected by potency?

Appears somewhat feral, he may have seen some domesticates, hemp for textiles and seed for food are fairly common place in china, from what im learning about it. I met a girl who came to the USA she was working to increase tourism to China while she was here in the states, I would like to see more strains from the desert oasis of NW China in seedbanks

 

[acespicoli]

Male Gametophyte (The Pollen Grain)​

The male gametophyte develops and reaches maturity in an immature anther. In a plant’s male reproductive organs, development of pollen takes place in a structure known as the microsporangium (Figure). The microsporangia, which are usually bilobed, are pollen sacs in which the microspores develop into pollen grains. These are found in the anther, which is at the end of the stamen—the long filament that supports the anther.

Shown is (a) a cross section of an anther at two developmental stages. The immature anther (top) contains four microsporangia, or pollen sacs. Each microsporangium contains hundreds of microspore mother cells that will each give rise to four pollen grains. The tapetum supports the development and maturation of the pollen grains. Upon maturation of the pollen (bottom), the pollen sac walls split open and the pollen grains (male gametophytes) are released, as shown in the (b) micrograph of an immature lily anther. In these scanning electron micrographs, pollen sacs are ready to burst, releasing their grains. (credit a: modification of work by LibreTexts; b: modification of work by Robert R. Wise; scale-bar data from Matt Russell)
Within the microsporangium, the microspore mother cell divides by meiosis to give rise to four microspores, each of which will ultimately form a pollen grain (Figure). An inner layer of cells, known as the tapetum, provides nutrition to the developing microspores and contributes key components to the pollen wall. Mature pollen grains contain two cells: a generative cell and a pollen tube cell. The generative cell is contained within the larger pollen tube cell. Upon germination, the tube cell forms the pollen tube through which the generative cell migrates to enter the ovary. During its transit inside the pollen tube, the generative cell divides to form two male gametes (sperm cells). Upon maturity, the microsporangia burst, releasing the pollen grains from the anther.

Pollen develops from the microspore mother cells. The mature pollen grain is composed of two cells: the pollen tube cell and the generative cell, which is inside the tube cell. The pollen grain has two coverings: an inner layer (intine) and an outer layer (exine). The inset scanning electron micrograph shows Arabidopsis lyrata pollen grains. (credit “pollen micrograph”: modification of work by Robert R. Wise; scale-bar data from Matt Russell)
Each pollen grain has two coverings: the exine (thicker, outer layer) and the intine (Figure). The exine contains sporopollenin, a complex waterproofing substance supplied by the tapetal cells. Sporopollenin allows the pollen to survive under unfavorable conditions and to be carried by wind, water, or biological agents without undergoing damage.

Female Gametophyte (The Embryo Sac)​

While the details may vary between species, the overall development of the female gametophyte has two distinct phases. First, in the process of megasporogenesis, a single cell in the diploid megasporangium—an area of tissue in the ovules—undergoes meiosis to produce four megaspores, only one of which survives. During the second phase, megagametogenesis, the surviving haploid megaspore undergoes mitosis to produce an eight-nucleate, seven-cell female gametophyte, also known as the megagametophyte or embryo sac. Two of the nuclei—the polar nuclei—move to the equator and fuse, forming a single, diploid central cell. This central cell later fuses with a sperm to form the triploid endosperm. Three nuclei position themselves on the end of the embryo sac opposite the micropyle and develop into the antipodal cells, which later degenerate. The nucleus closest to the micropyle becomes the female gamete, or egg cell, and the two adjacent nuclei develop into synergid cells (Figure). The synergids help guide the pollen tube for successful fertilization, after which they disintegrate. Once fertilization is complete, the resulting diploid zygote develops into the embryo, and the fertilized ovule forms the other tissues of the seed.

A double-layered integument protects the megasporangium and, later, the embryo sac. The integument will develop into the seed coat after fertilization and protect the entire seed. The ovule wall will become part of the fruit. The integuments, while protecting the megasporangium, do not enclose it completely, but leave an opening called the micropyle. The micropyle allows the pollen tube to enter the female gametophyte for fertilization.

Art Connection​

As shown in this diagram of the embryo sac in angiosperms, the ovule is covered by integuments and has an opening called a micropyle. Inside the embryo sac are three antipodal cells, two synergids, a central cell, and the egg cell.

Frontiers | Hermaphroditism in Marijuana (Cannabis sativa L.) Inflorescences – Impact on Floral Morphology, Seed Formation, Progeny Sex Ratios, and Genetic Variation

Cannabis sativa L. (hemp, marijuana) produces male and female inflorescences on different plants (dioecious) and therefore the plants are obligatory out-cros...

www.frontiersin.org

Cannabis sativa L. (hemp, marijuana) produces male and female inflorescences on different plants (dioecious) and therefore the plants are obligatory out-crossers. In commercial production, marijuana plants are all genetically female; male plants are destroyed as seed formation reduces flower quality. Spontaneously occurring hermaphroditic inflorescences, in which pistillate flowers are accompanied by formation of anthers, leads to undesired seed formation; the mechanism for this is poorly understood. We studied hermaphroditism in several marijuana strains with three objectives: (i) to compare the morphological features of this unique phenotype with normal male flowers; (ii) to assess pollen and seed viability from hermaphroditic flowers; and (iii) to assess the effect of hermaphroditism on progeny male:female (sex) ratios and on genetic variation using molecular methods. The morphological features of anthers, pollen production and germination in hermaphroditic flowers and in staminate inflorescences on male plants were compared using light and scanning electron microscopy. Seeds produced on hermaphroditic plants and seeds derived from cross-fertilization were germinated and seedlings were compared for gender ratios using a PCR-based assay as well as for the extent of genetic variation using six ISSR primers. Nei’s index of gene diversity and Shannon’s Information index were compared for these two populations. The morphology of anthers and pollen formation in hermaphroditic inflorescences was similar to that in staminate flowers. Seedlings from hermaphroditic seeds, and anther tissues, showed a female genetic composition while seedlings derived from cross-fertilized seeds showed a 1:1 male:female sex expression ratio. Uniquely, hermaphroditic inflorescences produced seeds which gave rise only to genetically female plants. In PCR assays, a 540 bp size fragment was present in male and female plants, while a 390 bp band was uniquely associated with male plants. Sequence analysis of these fragments revealed the presence of Copia-like retrotransposons within the C. sativa genome which may be associated with the expression of male or female phenotype. In ISSR analysis, the percentage of polymorphic loci ranged from 44 to 72% in hermaphroditic and cross-fertilized populations. Nei’s index of gene diversity and Shannon’s Information index were not statistically different for both populations. The extent of genetic variation after one generation of selfing in the progeny from hermaphroditic seed is similar to that in progeny from cross-fertilized seeds.

Preservation of male pollen and reversed female pollen ? XY&XX PRESERVATION ???

#pollentraders

http://cropgenebank.sgrp.cgiar.org/images/file/procedures/collecting1995/Chapter25.pdf

^^older paper must read^^

Anyone here ever thought about donating to a sperm bank ?
With samples of male and female reversed pollen ?
What conservation could we possibly achieve using these scientific techniques ?

 

[acespicoli]

• Cannabis sativa Click on organism name to get more information.
  • Cannabis sativa subsp. indica
  • Cannabis sativa subsp. sativa

 

[acespicoli]

• 
  
  
  Root system side view
  
  
• 
  
  
  Root system top view
  
  
• 
  
  
  Micrograph C. sativa (left), C. indica (right)
  
  

Genetics​

Cannabis, like many organisms, is diploid, having a chromosome complement of 2n=20, although polyploid individuals have been artificially produced.[43] The first genome sequence of Cannabis, which is estimated to be 820 Mb in size, was published in 2011 by a team of Canadian scientists.[44]

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Dwarfing - Wikipedia

en.wikipedia.org

Wild hemp sativa and cultivated indica and example of agricultural dwarfism ?

 

[acespicoli]

https://mdpi-res.com/d_attachment/plants/plants-09-00665/article_deploy/plants-09-00665-with-cover.pdf?version=1669136278

 

[acespicoli]

Genetic data from over 1,300 cannabis varieties is now available for public access.

When our customers give us permission, Phylos shares a set of raw genetic data from their cannabis varieties with the National Center for Biotechnology Information (NCBI), one of the world’s largest repositories of open access genetic data.

phylos.bio

The power of Open Data​

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Dec 18, 2018

Genetic data from over 1,300 cannabis varieties is now available for public access.​

Phylos reads the DNA of individual cannabis varieties using next generation sequencing and high density genotyping technology. During this process, we generate data, analyze it, and compare samples to each other, which makes it possible to produce genetic reports and assign varieties an exact location in the Phylos Galaxy.
When analyzed in aggregate, genotype data serve as an extremely powerful tool for advancing cannabis science. Genotype data can be used to understand relationships between varieties, the structure of populations, and to deepen our understanding of how genetic variation determines the phenotypes we see in different varieties. It also serves as a valuable resource for other researchers to supplement and expand their own studies.
As a science company, we believe it is our responsibility to increase the amount of publicly shared data available to both the research community and cannabis industry. Making data publicly accessible is a well established practice.
When our customers give us permission, Phylos shares a set of raw genetic data from their cannabis varieties with the National Center for Biotechnology Information (NCBI), one of the world’s largest repositories of open access genetic data.
In 2016, Phylos published approximately 850 raw genetic sequences to the NCBI. Earlier this week, we published a second set with data from over 1,300 varieties. Once data is published to the NCBI (or other public data repositories), anyone can access, view, and download it. Varied researchers and nonprofits have accessed this data, from Google Cloud’s BigQuery to the Open Cannabis Project—a nonprofit organization with a mission to defend cannabis diversity from overreaching patents.
While this DNA sequence data is extremely valuable from a research perspective, it is important to note that it cannot be used to grow plants, to claim ownership over them, or to patent them. Thankfully, public access to data does not weaken an owner’s rights to pursue intellectual property on an individual plant, nor does it place the plant into the public domain unless clonally propagated cuttings have been previously released.
Phylos customers can use their sequence data as a form of intellectual property documentation and protection, although genetic data alone does not provide ownership rights for a plant. However, genetic data can be used defensively as evidence to stop other parties from filing illegitimate protections on a plant or category of plants, and offensively as supporting evidence of an owner’s intellectual property.
Download Phylos data.
Contact [email protected] for more information on Phylos data sharing and privacy.

phylos bioscience - BioSample - NCBI

phylos bioscience panama - BioSample - NCBI

phylos bioscience colombian gold - BioSample - NCBI

phylos bioscience oaxaca - BioSample - NCBI

753cfbd4814c-TSCA_Romulan - BioSample - NCBI

Genome

Download a genome data package including genome, transcript and protein sequence, annotation and a data report

www.ncbi.nlm.nih.gov

Large-scale whole-genome resequencing unravels the domestication history of Cannabis sativa < linked​

• A high-quality reference genome of wild Cannabis sativa. Gao S, et al. Hortic Res 2020
• A physical and genetic map of Cannabis sativa identifies extensive rearrangements at the THC/CBD acid synthase loci. Laverty KU, et al. Genome Res 2019 Jan
• Two complete chloroplast genome sequences of Cannabis sativa varieties. Oh H, et al. Mitochondrial DNA A DNA Mapp Seq Anal 2016 Jul
• The draft genome and transcriptome of Cannabis sativa. van Bakel H, et al. Genome Biol 2011 Oct 20

I find it interesting that vinifera (grape .ie. wine) somniferum (opium poppy) and cannabis all branch relatively closely. Many interests in these articles. One is to preserve landrace genetics the other is to provide much needed medical cures and pain treatment to people in need. Many of these plants have been saved thru generations for their therapeutic properties. One chemical in a pill is great when it works, the synergies of the natural balances of tepenes and cannabinoids cant be dismissed as a complete medicine. While one chemical may treat a symptom maybe it is the thing as a whole that is the cure.

So often today I see doctors treat the symptoms and not cure the underlying disease...
In the USA doctors are becoming a rareity your more likey to have a symptom treated by a nurse
There is hope for a better healer one with a broader view to cure disease and no longer need a medicine for the symptom...

Maybe not a great idea in the view of a drug company?

This from phylos what they offered in the past...

Genotype data can be used to understand relationships between varieties, the structure of populations, and to deepen our understanding of how genetic variation determines the phenotypes we see in different varieties. It also serves as a valuable resource for other researchers to supplement and expand their own studies....

None the less we need a better way to view use the available data than looking at shotgun arrays

Shotgun sequencing - Wikipedia

en.wikipedia.org

Discontinuation of the Phylos Plant Sex Test and Genotype Test​

The Phylos Plant Sex Test and Genotype Test are no longer accepted for processing as of September 30, 2021. Full customer account access, sex test results, and genotype reports will all remain available through our website for the foreseeable future.​

Due to the complexity of the 3D Galaxy data visualization tool and the discontinuation of testing services and related resources, the interactive Phylos Galaxy is no longer available to explore. For an example of the interactive 3D Galaxy, check out our videos on YouTube.

Final Galaxy dataset upload​

Our latest (and last) Galaxy public dataset has been published in the European Bioinformatics Institute (EMBL-EBI) archives. The European Variation Archive (EVA) is an open-access database of all types of genetic variation data from all species.

We believe the foundational knowledge of the cannabis genome can be advanced and better understood by sharing this incredible data we have collected for the Galaxy. To download all public Phylos-collected genotype data, visit EBI’s archive.

The European Bioinformatics Institute < EMBL-EBI

EMBL-EBI

www.ebi.ac.uk

https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7195422/pdf/41438_2020_Article_295.pdf

very good read they mention some good ways to use the data sets

GitHub - mahulchak/quickmerge: A simple and fast metassembler and assembly gap filler designed for long molecule based assemblies.

A simple and fast metassembler and assembly gap filler designed for long molecule based assemblies. - mahulchak/quickmerge

github.com

So there is the software and also we need the dna testing

 

[acespicoli]

Ernest Small, Arthur Cronquist

https://www.researchgate.net/profile/Robert-Clarke-2

https://www.researchgate.net/profile/Mark-Merlin

International Hemp Association

Dedicated to the advancement of Cannabis through the dissemination of information.

www.internationalhempassociation.org

I would like to see a public repository of genetic data
The things of past/present projects are valuable listing a few below

Seedfinder

Phylos
Genotype data can be used to understand relationships between varieties, the structure of populations, and to deepen our understanding of how genetic variation determines the phenotypes we see in different varieties. It also serves as a valuable resource for other researchers to supplement and expand their own studies....

Leafly

Contiguous and accurate de novo assembly of metazoan genomes with modest long read coverage

Abstract. Genome assemblies that are accurate, complete and contiguous are essential for identifying important structural and functional elements of genome

academic.oup.com

Published online 25 July 2016 Nucleic Acids Research, 2016, Vol. 44, No. 19
pdf link

Yeah so each site had/has its merits and would be valuable in a future database for breeders medical users smokers, so what would we catalog why and how ? Feel free to add more data sources and examples as well as ideas etc

 

[acespicoli]

Nationwide Hemp Testing Lab​

Modern Canna is proud to be the only certified Leafly partner in Florida.
We only work with cannabis and hemp companies who take their respective industries seriously and who aim to build sustainable businesses. For hemp clients, the component of interest is almost always going to be cannabidiol (CBD).
Quality control testing is imperative for hemp growers and hemp-derived product manufacturers. Whether for medical or recreational use, users have a right to know exactly what they’re getting. Testing with an accredited third party laboratory like Modern Canna establishes credibility as a supplier and instills consumer trust. Quality control is of increasing importance as the hemp industry continues to grow.
Modern Canna provides a full range of testing services geared towards analyzing hemp and hemp-derived samples for contaminants. These services include, but are not limited to, pesticides, heavy metals, mycotoxins, microbials, and residual solvents. Furthermore, we analyze samples for their potency, which includes a full range of cannabanoids, most importantly the psychoactive compound, THC.

How the Hemp Testing Process Works​

1.) Find a good laboratory – Find a laboratory that is not only ISO/IEC-17025 certified, but make sure they also provide proof of how they validate their data. For more on this see our Quality Control page.
2.) Complete a Chain of Custody (COC) form – The COC is used for sample submission and helps the laboratory keep track of the entire hemp testing process. This form contains information pertaining to sample material, sample weight, analyses required, and sample collection date and time. The COC also keeps track of each entity handling the hemp materials, from sample collection, to transportation, to the time lab results are reported out.
3.) Sample transportation – Coordinate with the laboratory on whether to ship samples or request a courier.
4.) Wait for results – This is where the lab takes over. The hemp lab testing process consists of the following:
A.) Once the samples arrive to the lab they get logged into the laboratory information management system (LIMS).
B.) Samples are then prepped. Sample preparation is tedious and requires skilled analysts and sound methodology. Some examples of sample preparation are weighing, grinding, pulverizing, flash-freezing, diluting, extracting, digesting, sonicating, and filtering.
C.) Once the samples are prepped they are ran on an instrument specific to the analysis required. Sample run times vary depending on the analyses required.
D.) After samples are run the data is reviewed and thoroughly evaluated by lead analysts and lab managers.
E.) Lastly the data is converted into full report and certificate of analysis (COA) formats then sent to the clients.

What We Test For​

The passing of the Farm Bill in 2018 was a major milestone that legalized the commercial production of hemp. The federal law is applicable in every state. There is a caveat, however; the hemp material cannot exceed 0.3% THC concentrations.
Without testing before product release, hemp cultivators and processors risk serious legal liability. The risk extends beyond THC concentrations. Hemp must also contain absolute minimal traces of other impurities.
At Modern Canna, we test for cannabinoids like CBD and THC, as well as the following:

• Terpenes – There are an estimated 200 terpenes found in hemp. We test for the most common terpenes, which are essential for the hemp’s aroma and botanical effects.
• Residual solvents – Solvents are needed to extract cannabinoids from the raw plant. However, residual solvents left behind include contaminants like butane and propane.
• Moisture content – We use a drying method or moisture balance instrument to test for a hemp sample’s water weight.
• Heavy metals – Heavy metals can pass onto the hemp from contaminated soil/water or vape pens. Modern Canna tests for harmful metals like lead, mercury, cadmium, lead, etc.
• Microbial impurities – Detecting and identifying harmful microbial content. This includes those found in the raw plant, such as mold, yeast, aspergillus, and E. Coli.
• Pesticides – Pesticides are a part of hemp cultivation, just as with any other crops. We test for pesticide residue that includes insecticides, fungicides, and plant growth regulators.
• Water activity – This is not to be confused with moisture content. Water activity testing assesses moisture levels on the plant’s surface, which cannot surpass certain concentrations. Excess moisture elevates the risk of microbial growth.

Entrust Hemp Testing to Modern Canna​

There is a reason why Modern Canna was selected as one of the first initial labs in the United States for Leafly’s certified lab program. We provide timely, comprehensive, and accurate testing to ensure hemp product are safe and meet the most stringent regulations. Contact us today to learn more about our hemp testing services. We look forward to working with you.


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