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Old 02-28-2018, 10:21 PM #31
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( Part - 1 )

Cannabis, an erect annual herb and member of the Cannabaceae family, is monotypic and characterized by a single species Cannabis sativa L. (Small and Cronquist, 1976). Plants are diploid (2n = 20) with an estimated haploid genome of ~830 Mb (Van Bakel et al., 2011). The extant genepool is thought to be comprised primarily of domesticated or feral populations, cultivars, and selections (Small and Cronquist, 1976), with a subset having been subject to steep selective gradients toward phenotypes for specific end-uses (Mandolino and Carboni, 2004; Potter, 2014; Small, 2015a). Cannabis has been cultivated in Eurasia over several thousand years (Li, 1974; Bradshaw et al., 1981; Murphy et al., 2011; Herbig and Sirocko, 2013) and has since radiated from this region and been subject to prolonged artificial selective pressures in Africa (Duvall, 2016) and North and South America (Small and Marcus, 2002), and is now cultivated globally (Salentijn et al., 2014). Plants are diecious and obligate outbred, although some fiber forms are monecious (Faeti et al., 1996). This has contributed to a high level of hybridization between pre-, post-, and de-domesticated populations (Gilmore et al., 2007), and therefore few if any intact wild populations are thought to exist (Small and Cronquist, 1976).

Transcriptome analysis (Van Bakel et al., 2011; McKernan et al., 2015; Onofri et al., 2015; Weiblen et al., 2015) and QTL mapping (Weiblen et al., 2015) further support the historical selection pressure that has led to chemotypic separation between contemporary fiber and marijuana groupings. Cannabinoids accumulate in plants in their carboxylic acid forms, such as tetrahydrocannabinolic acid (THCA) and cannabidiolic acid (CBDA) which form neutral cannabinoids THC and CBD in a non-enzymatic reaction when exposed to heat (Dussy et al., 2005). The marijuana variety Purple Kush was found to have higher expression levels of genes encoding cannabinoid biosynthetic pathway intermediates than the fiber cultivar Finola, with Purple Kush only expressing functional sequence variants of genes coding for THCA and Finola those for CBDA synthase (Van Bakel et al., 2011). Linkage mapping in 62 F2 individuals derived from a cross between full-sib inbred contemporary fiber cultivar Carmen and marijuana variety Skunk#1 revealed QTL for THCA and CBDA composition, as well as putative QTL for cannabinoid content, with differences in composition associated with a CBDA synthase locus and loss of CBDA synthase functionality in Skunk#1 (Weiblen et al., 2015).

The occurrence of contemporary fiber cultivars which have relatively high levels of CBDA (Small, 2015a) can be attributed to breeding efforts within France and other European countries toward the middle to latter part of the twentieth century (Amaducci et al., 2014), where techniques such as the Bredemann method, an in vivo fiber evaluation method, and the counter selection for THC using marker assisted selection (MAS), were employed (Ranalli, 2004). Fifty one fiber cultivars are currently registered for use within the European Union (Salentijn et al., 2014) and these cultivars have been exported to North America (Small, 2015a) and the Northern provinces of China (Salentijn et al., 2014). Despite the scarcity of published data relating to ancestry of such accessions, it is believed that a large number of contemporary fiber cultivars are descendants from Central Russian and Mediterranean landraces and derivative cross-progenies (De Meijer and van Soest, 1992).

Phylogenetic relationships between domesticated Cannabis germplasm have recently been examined using reduced representation DNA sequencing. Genotyping by sequencing (GBS) analysis of 195 accessions using 2894 single nucleotide polymorphisms (SNPs) inferred close relatedness and shared ancestry between contemporary fiber accessions, with the latter forming a separate clade (Lynch et al., 2015). These accessions were also observed to exhibit lower levels of heterozygosity than other intraspecific taxa (Mann-Whitney U-test p < 0.001; Lynch et al., 2015), suggesting that recent domestication of fiber traits has resulted in a genetic bottleneck and reduction in allelic diversity. However, these varieties were not well-represented in the sample collection, with only 16 analyzed (Lynch et al., 2015). Moreover, a separate GBS study using 14,031 SNPs in 43 contemporary fiber and 81 marijuana varieties produced results that conflicted with this more recent study, and showed significantly lower levels of heterozygosity in marijuana varieties compared with fiber cultivars (Mann-Whitney U-test p = 8.64 × 10−14; Sawler et al., 2015).

Despite this lack of congruence between GBS analyses, domestication for either industrial hemp or marijuana traits has likely resulted in a loss of genetic and allelic diversity, potentially brought about by changes in breeding systems. Processes such as linkage drag can be associated with complex polygenic flowering QTL (Mace et al., 2013) in relation to latitudinal and environmental adaption (Gao et al., 2014). Regardless of reductions in allelic diversity that may have arisen either from clonal propagation in marijuana (Russo, 2007), or from the propagation of monoecious varieties in industrial hemp (Forapani et al., 2001), it is unclear to what extent contemporary Cannabis germplasm deviates from the broader genepool. Analysis of 45 SNPs in both GBS sample sets reveals an overall limited genetic distance between 22 industrial hemp and 173 marijuana groupings (Lynch et al., 2015). Resequencing and mapping of 30 billion sequence reads from 302 domesticated and wild soybean (Glycine max) accessions identified selective sweeps associated with domestication events (Zhou et al., 2015).
By comparison, resequencing of various species of Citrus has also revealed a complex arrangement of large haplotype blocks and admixture between ancestral and domesticated species (Wu et al., 2014). On completion of a fully annotated Cannabis genome (Van Bakel et al., 2011), it may be possible to resequence diverse germplasm (Scossa et al., 2015) to quantify differences in genetic diversity and to determine the contribution wild ancestors have conferred to contemporary forms. However, access to wild and landrace accessions may be a limiting factor to exploring Cannabis phylogeny.

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Old 02-28-2018, 10:22 PM #32
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( Part - 2 )

The region associated with the origin of a species is often considered the center of genetic diversity, although this may not be the case with species that have been subject to prolonged periods of domestication and secondary radiation. This has been observed in the common bean (Phaseolus vulgaris), where simple sequence repeat (SSR) genetic marker comparisons between native American germplasm and secondary domesticated groupings revealed a higher level of nuclear diversity in the African germplasm than that of the native collection, suggesting that non-native geographical regions can act as both source and sinks for genetic diversity over different historical periods (Bellucci et al., 2014). Palaeobotanical evidence in the form of pollen deposits and historical accounts would strongly suggest domestication of Cannabis in the post-Neolithic era over tens of thousands of years, with subsequent secondary domestication events in non-native geographical regions (Li, 1974; Bradshaw et al., 1981; Small and Marcus, 2002; Murphy et al., 2011; Herbig and Sirocko, 2013; Duvall, 2016).

Based on field observations it has been concluded that Cannabis originated within Central Asia (Hillig, 2005b; Russo, 2007) and references therein, although such inferences may not be justified given the domestication and radiation of Cannabis throughout Eurasia over several millennia (Li, 1974; Bradshaw et al., 1981; Murphy et al., 2011; Herbig and Sirocko, 2013). Conclusive phylogenetic evidence in support of a specific geographical region is also incomplete, with ruderal or wild populations underrepresented or absent from sample collections (Hillig, 2005b; Lynch et al., 2015). Combining genetic and phenotypic evidence relating to the predominant characteristics of a species in both ruderal and domesticated forms in the context of their allelic eco-geographical distribution, is more likely to define a putative center of diversity, as has been noted in other widely cultivated plant species (Smith and King, 2000).

East Asia appears to be a rich source of genetic diversity within the Cannabis genepool, and a potentially valuable genetic resource both for future phylogenetic analyses and ex situ conservation. China is botanically megadiverse (Li, 2008). The Hengduan Mountains in the south west of China have been identified as one of only 35 biodiversity hotspots worldwide (Sloan et al., 2014) and this region encompasses parts of the Yunnan province in which the Yunnan Academy of Agricultural Sciences (YAAS) Cannabis germplasm collection is maintained. China also benefits from a latitudinal gradient from ~23–50°N (Amaducci et al., 2014), with hundreds of Cannabis landraces reported to have undergone distinct domestication events along these latitudes in provinces spanning from Hebei in the north west, through to ShanDong, Henan, Guizhou and Yunnan in the south west (Salentijn et al., 2014). Moreover, historical evidence strongly suggests that Cannabis has been cultivated in China over several thousand years. Pottery paintings depicting Cannabis are believed to have been produced by the Neolithic Yangshao culture, and Cannabis fibers were reportedly utilized in the production of paper during the Han dynasty >1790 years before present (Li, 1974). Excavation of the 2700 year old Yanghai Tombs in Xinjiang-Uyghur Autonomous Region in China has also revealed high THC plant material (Russo et al., 2008), implying that this plant was used within a cultural and potentially medicinal context within early Chinese societies.

A number of genetic markers have been used to determine levels of heterozygosity in Chinese Cannabis germplasm (Table 1), with SSR, amplified fragment length polymorphism (AFLP) and randomly amplified polymorphic DNA (RAPD) genetic markers indicating a high level of genetic diversity, with the proportion of polymorphic loci ranging from 75 to 92% (Gilmore et al., 2007; Hu et al., 2012; Gao et al., 2014; Zhang et al., 2014). Analysis of 76 accessions from 26 countries using 12 chloroplast and mitochondrial DNA loci revealed six haplotypes, all of which were located within or adjacent to China (Gilmore et al., 2007). Fifty six loci derived from expressed sequence tag (EST) simple sequence repeat (EST-SSR) markers were tested on a sample collection of 100 varieties from 10 provinces in China and 15 varieties from Europe. Principle coordinate analysis revealed four clusters relating to geographical location and latitude. Interestingly, clusters from Central, Northern, and Southern China had a higher percentage of polymorphic loci than the European cluster (Gao et al., 2014; Table 1), suggesting a higher level of diversity within Chinese germplasm.

Of the 808 accessions reported to have been collected within ex situ Cannabis genetic resource collections worldwide, only 58 (7.2%) have origins within China (Figure 3). Moreover, from the 156 accessions listed in the former Centre for Plant Breeding and Reproduction Research (CPRO) germplasm collection, cultivars of predominantly European origin and marijuana varieties contributed to more than 40% of all accessions (De Meijer and van Soest, 1992). Considering the long cultivation history of Cannabis in China, and the high density of landrace accessions occurring within a large latitudinal range, sourcing accessions from China and adjacent regions should be a priority for Cannabis ex situ conservation, irrespective of whether the center of origin for this species has been fully characterized. Nevertheless, appropriate management of germplasm ex situ and systematic characterization of East Asian Cannabis genetic resources will be required if the full potential of genepool-enabled crop improvement is to be maximized.



https://www.frontiersin.org/articles...016.01113/full
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Old 02-28-2018, 10:32 PM #33
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( Part - 3 )

Regeneration is one of the most costly factors in the conservation of ex situ genetic resource collections and requires significant input in the form of labor, resources, and infrastructure (Li and Pritchard, 2009; Khoury et al., 2010). The regeneration of Cannabis germplasm is especially problematic as plants are anemophilous (wind pollinated) and dioecious, with male or hermaphrodite plants releasing large amounts of pollen during flowering (Amaducci et al., 2014). It has been estimated that a single plant can produce in excess of 350 million pollen grains (Faegri et al., 1989). Prevention of gene flow and hybridization between accessions is therefore an important consideration in Cannabis ex situ regeneration and conservation (De Meijer and van Soest, 1992), and it may have contributed to reports of a widely shared Cannabis genepool.

Outdoor regeneration can require large areas of land, with distances of up to 5 km required to prevent cross-pollination (De Meijer and van Soest, 1992; Small and Antle, 2003). However, given the long distances Cannabis pollen can travel and the sensitivity of pollen distribution to wind velocity (Small and Antle, 2003), as well as the potential for prolonged viability of Cannabis pollen in low relative humidities post-anthesis (Bassani et al., 1994), introgression may not necessarily be prevented at these distances. Outdoor regeneration can also be impractical where multiplication of diverse accessions requires specific photoperiods spanning several degrees of latitude (Gao et al., 2014) in order to initiate flowering (Cosentino et al., 2012), thus limiting multiplication of Cannabis germplasm to certain periods throughout the year.

Protected cultivation within pollen secure facilities is a standard alternative to outdoor multiplication. However, these may be costly and also problematic in maintaining genetic diversity. For many plant species, genetic drift has been attributed to the process of regeneration, and associations have been detected between time spent ex situ and loss of alleles per locus, gene diversity, and percentage of polymorphic loci (Parzies et al., 2000). For example, loss of ex situ genetic diversity has been observed in outcrossing species such as barley (Hordeum vulgare L.), maize and common bean genetic resource collections (Parzies et al., 2000; Rice et al., 2006; Negri and Tiranti, 2010), although much of the reduction in genetic diversity appears dependent on the number of parents used in mass pollination (Parzies et al., 2000). Given the spatial limitations associated with the construction of indoor pollen-secure infrastructure (Negri and Tiranti, 2010), which limits the number of outbreeding individuals for regeneration, a decline in allelic variation may occur with each regeneration cycle, ultimately leading to erosion of genetic variability from the time of acquisition.

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Old 03-01-2018, 01:36 AM #34
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Lyster Dewey with 'kymington' hemp standing where the pentgon now sits. From Lyster Dewey diaries found at a garage sale...
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Old 03-01-2018, 08:35 AM #35
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Originally Posted by Kankakee View Post
( Part - 3 )

Regeneration is one of the most costly factors in the conservation of ex situ genetic resource collections and requires significant input in the form of labor, resources, and infrastructure (Li and Pritchard, 2009; Khoury et al., 2010). The regeneration of Cannabis germplasm is especially problematic as plants are anemophilous (wind pollinated) and dioecious, with male or hermaphrodite plants releasing large amounts of pollen during flowering (Amaducci et al., 2014). It has been estimated that a single plant can produce in excess of 350 million pollen grains (Faegri et al., 1989). Prevention of gene flow and hybridization between accessions is therefore an important consideration in Cannabis ex situ regeneration and conservation (De Meijer and van Soest, 1992), and it may have contributed to reports of a widely shared Cannabis genepool.

Outdoor regeneration can require large areas of land, with distances of up to 5 km required to prevent cross-pollination (De Meijer and van Soest, 1992; Small and Antle, 2003). However, given the long distances Cannabis pollen can travel and the sensitivity of pollen distribution to wind velocity (Small and Antle, 2003), as well as the potential for prolonged viability of Cannabis pollen in low relative humidities post-anthesis (Bassani et al., 1994), introgression may not necessarily be prevented at these distances. Outdoor regeneration can also be impractical where multiplication of diverse accessions requires specific photoperiods spanning several degrees of latitude (Gao et al., 2014) in order to initiate flowering (Cosentino et al., 2012), thus limiting multiplication of Cannabis germplasm to certain periods throughout the year.

Protected cultivation within pollen secure facilities is a standard alternative to outdoor multiplication. However, these may be costly and also problematic in maintaining genetic diversity. For many plant species, genetic drift has been attributed to the process of regeneration, and associations have been detected between time spent ex situ and loss of alleles per locus, gene diversity, and percentage of polymorphic loci (Parzies et al., 2000). For example, loss of ex situ genetic diversity has been observed in outcrossing species such as barley (Hordeum vulgare L.), maize and common bean genetic resource collections (Parzies et al., 2000; Rice et al., 2006; Negri and Tiranti, 2010), although much of the reduction in genetic diversity appears dependent on the number of parents used in mass pollination (Parzies et al., 2000). Given the spatial limitations associated with the construction of indoor pollen-secure infrastructure (Negri and Tiranti, 2010), which limits the number of outbreeding individuals for regeneration, a decline in allelic variation may occur with each regeneration cycle, ultimately leading to erosion of genetic variability from the time of acquisition.
Quote:
Signs in Wind of Morocco Drug Crop
By MARLISE SIMONS
Published: June 18, 1995

MADRID— Scientists sampling the air in southern Spain the other day came across a surprising event, a great stream of marijuana pollen coming off the Mediterranean waters, carried by a warm southern wind.

The pollen, though invisible to the naked eye, was measured along no less than a 250-mile stretch of the Spanish coast, from Estepona to Cartagena, and it reached more than 100 miles inland, beyond Cordoba.

"This is exceptional," said Eugenio Dominguez, coordinator of Spain's Network for Aerobiology, which detected the particles. "We've never measured marijuana pollen in so many places."

Researchers soon established that the tiny grains appearing in their microscopes were harbingers of a likely bumper crop of marijuana in Morocco, across the water some 25 miles to the south. At this time of year, they said, the great marijuana plantations are in flower along the north coast of Morocco between Tangiers and the Algerian border.
https://www.nytimes.com/1995/06/18/world/signs-in-wind-of-morocco-drug-crop.html
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Old 03-01-2018, 04:23 PM #36
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The spot I'll be using for bulking fiber line is situated within a 250 acre state forest preserve untouched for hundreds of years already teaming with worm casting in some of the best black soil on earth. The area is a small 4 acre opening that is also surrounded by millions of 12-14 ft. stinging nettle's acres upon acres that will make everything indistinguishable from ten feet let alone one hundred or from above.

Having grown in this spot for many years and never having pollen contamination issue's is very important.

The nettles grow at very high density and when inside that area everything turns black. The photo below was taken at mid day and other photo's show the area in the spring before nettles composting downwards towards the ground..
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Old 03-01-2018, 04:29 PM #37
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Lyster Dewey at Arlington farms 1929. The fiber line I've found match this plant structure as the branching started five to seven feet above ground and very limited. Also a sativa flower set structure limiting seed production.
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Old 03-01-2018, 04:57 PM #38
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Lyster Dewey at Arlington farms 1929. The fiber line I've found match this plant structure as the branching started five to seven feet above ground and very limited. Also a sativa flower set structure limiting seed production.
Interesting.Did you also take look to the Hmong?
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Old 03-01-2018, 09:25 PM #39
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No what is Hmong ?

And the hemp line I'll be using for outcrossing in future is not from China today but borders China. Its a private line but derived from those genetics....
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Old 03-01-2018, 10:03 PM #40
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Then you must know the Hmong people from Laos.

https://www.passa-paa.com/the-hmong-hemp-influence/
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