Novel polymorphic microsatellite markers in section Caulorrhizae (Arachis,Fabaceae)

This work reports the characterization of 11 polymorphic microsatellite loci in section Caulorrhizae. The primer pairs were designed from Arachis pintoi and showed full transferability to Arachis repens species. These new markers were used to evaluate the genetic diversity in germplasm (accessions and cultivars) of section Caulorrhizae. This new set of markers detected greater gene diversity than morphological and molecular markers such as AFLP (amplified fragment length polymorphism) and RAPD (rapid analysis of polymorphic DNA) previously used in this germplasm.     

Phytochemical Characterization of Cannabis sativa L. Roots from Northeastern Brazil

This article describes the phytochemical study of Cannabis sativa roots from northeastern Brazil. The dried plant material was pulverized and subjected to exhaustive maceration with ethanol at room temperature, obtaining the crude ethanolic extract (Cs-EEBR). The volatile compounds were analyzed by gas chromatography coupled with mass spectrometry (GC/MS), which allowed to identify 22 compounds by comparing the linear retention index (LRI), the similarity index (SI) and the fragmentation pattern of the constituents with the literature. By this technique the major compounds identified were: friedelan-3-one and β-sitosterol. In addition, two fractions were obtained from Cs-EEBR by classical column chromatography and preparative thin layer chromatography. These fractions were analyzed by NMR and IR and together with the mass spectrometry data allowed to identify the compounds: epifriedelanol, friedelan-3-one, β-sitosterol and stigmasterol. The study contributed to the phytochemical knowledge of Cannabis sativa, specifically the roots, as there are few reports on the chemical constituents of this part of the plant.     

Tetrahydrocannabinol levels in hemp (cannabis sativa) germplasm resources

In most of the western world where industrial hemp, Cannabis sativa, is licensed for cultivation, the plants must not exceed a level of 0.3% tetrahydrocannabinol (THC), the principal intoxicating constituent of the species. Because there are no publicly available germplasm hemp collections in North America and only a very few, recent North American cultivars have been bred, the future breeding of cultivars suitable for North America is heavily dependent on European cultivars and European germplasm collections. Based mostly on material from Europe, this study surveyed THC levels of 167 accessions grown in southern Ontario, making this the largest survey to date of germplasm intended for breeding in North America. Forty-three percent of these had THC levels ≥0.3% and, therefore, are unsuitable for hemp development in North America. Discrepancies were found between THC levels reported for some germplasm holdings in Europe when they were grown in Canada and, accordingly, verification of THC levels developed in North America is necessary.     

大麻染色体行为分析

以大麻不同性别的植株为材料,常规压片法观察细胞染色体行为规律。核型分析结果表明:大麻雌雄株的体细胞染色体数目均为2n=20,核型公式分别为雌株2n=2x=20=18m 2sm,雄株2n=2x=20=18m 2sm(1SAT)。雌株体细胞中有2条X染色体,而雄株只有一条X染色体和一条具有大随体的Y染色体。雌雄株核型均为2A型,为较对称核型。这一结果可为进一步研究大麻性别的分化机制提供细胞遗传学理论依据。     

Isolation and characterization of microsatellite markers from Robinia pseudoacacia L

Microsatellite markers were isolated from Robinia pseudoacacia L. using an enrichment method. Eleven of the 23 primer pairs designed successfully amplified unambiguous and polymorphic single loci among 39 individual R. pseudoacacia L. from northeastern Japan. The observed and expected heterozygosities of the 11 microsatellite markers ranged from 0.333 to 0.821 and from 0.489 to 0.867, respectively. The polymorphisms observed at the 11 microsatellite loci are useful genetic data for forest ecological studies involving R. pseudoacacia L.     

The effect of temperature on leaf appearance and canopy establishment in fibre hemp (Cannabis sativa L.)

The effects of temperature on the development and growth of hemp (Cannabis sativa L.) have never been quantified. Therefore, to establish the effect of temperature on leaf appearance and canopy establishment of fibre hemp under controlled and field conditions, plants were grown in growth chambers at 11 regimes with average temperatures between 10°C and 28°C, and three cultivars were sown in the field in March, April and May in 1990, 1991 and 1992. In the field, thermal time (base 0°C) between sowing and emergence ranged from 68°Cd to 109.5°Cd (average 88.3°Cd). Rates of leaf appearance and stem elongation increased linearly with temperature between 10°C and 28°C. The base temperature for leaf appearance was 5.7°C from the growth chamber experiments and 1°C from the field experiments. In the field, the base temperature for the relationship between light interception by the canopy and thermal time was 2.5°C, and thermal time, calculated at the appropriate base temperature, accounted for about 98% of the variance in the number of leaves and for 98.6% of the variance in the proportion of light intercepted by the canopy. Days from emergence accounted for less of the variance in both parameters than thermal time. Interception of 90% of light was attained on average at 465°Cd (base 0°C) after emergence. It is concluded that thermal time is a simple and accurate tool to describe leaf appearance and light interception in fibre hemp.     

Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.)

 A 400-bp RAPD marker generated by a primer of random decamer sequence has been found associated with the male sex phenotype in 14 dioecious cultivars and accessions of hemp (Cannabis sativa L.). The primer OPA8 generates a set of bands, most of which polymorphic among all the individual plants tested, and 1 of which, named OPA8₄₀₀, present in all male plants and absent in female plants. A screening of 167 plants belonging to different genotypes for the association of the OPA8₄₀₀ marker with the sex phenotype revealed that only in 3 cases was the 400-bp band was present in plants phenotypically female; on the contrary, in male plants the band was never missing, while in monoecious plants it was never present. Despite this sex-specific association, the sequences corresponding to OPA8₄₀₀ were present in both staminate and carpellate plants, as revealed by Southern blotting and hybridization with the cloned RAPD band. The RAPD marker was sequenced, and specific primers were constructed. These primers generated, on the same genotypes used for RAPD analysis, a SCAR marker 390 bp in length and male-specific. This SCAR is suitable for a precise, early and rapid identification of male plants during breeding programs of dioecious and monoecious hemp. Received: 16 January 1998 / Accepted: 30 April 1998     

Zur virusanfälligkeit von hanfsorten (Cannabis sativa L.)

Der Anbau von Hanf (Cannabis sativa L.) als nachwachsender Rohstoff nimmt in Europa zu. Untersuchungen zur Anfälligkeit von drei Genotypen gegenüber 8 phytopathogenen Viren aus unterschiedlichen taxonomischen Gruppen zeigten deutliche Unterschiede.     

Genetics and Marker-assisted Selection of the Chemotype in Cannabis sativa L.

Cannabis sativa is an interesting crop for several industrial uses, but the legislations in Europe and USA require a tight control of cannabinoid type and content for cultivation and subsidies release. Therefore, cannabinoid survey by gas chromatography of materials under selection is an important step in hemp breeding. In this paper, a number of Cannabis accessions were examined for their cannabinoid composition. Their absolute and relative content was examined, and results are discussed in the light of both the current genetic model for cannabinoid’s inheritance, and the legislation’s requirements. In addition, the effectiveness of two different types of markers associated to the locus determining the chemotype in Cannabis was evaluated and discussed, as possible tools in marker-assisted selection in hemp, but also for possible applications in the forensic and pharmaceutical fields.     

Isolation and characterization of microsatellite markers in the South African abalone (Haliotis midae)

We report the isolation and characterization of 11 polymorphic microsatellite loci in the South African abalone Haliotis midae. These loci showed a range of five to 21 alleles per locus and observed heterozygosities ranging from 0.14 to 0.93 in a wild population of 32 individuals. All loci except four conformed to Hardy–Weinberg expectations and did not show linkage disequilibrium. The polymorphism exhibited at these loci indicate that they would be useful in determining levels of genetic variability in natural and commercial Haliotis midae populations as well as in parentage and Quantitative Trait Loci (QTL) analysis in hatchery reared abalone.     

Conditioning of Heavy Metal-Polluted River Sediment by Cannabis sativa L.

We are currently developing a process for the remediation of heavy metal-polluted sediment by solid-bed bioleaching. Unfortunately, freshly dredged sediment is impermeable to water, unsuitable for solid-bed leaching, and therefore needs preliminary conditioning to improve its structure. A pilot-scale conditioning plant consisting of several basins (each with a base of 6 m² and a depth of 0.7 m) was filled with freshly dredged, slightly predewatered sediment from the Weisse Elster River. Hemp (Cannabis sativa L.) was chosen for conditioning because it develops quickly from seeds, produces large amounts of biomass, and forms commercially useful fibers. Irrespective of the plant cover, the sediment dried out, was oxidized and turned acidic. However, its structure and permeability only improved when the sediment was covered with plants and not when all the vegetation appearing was removed regularly. Although hemp germinated completely regularly on sediment, 1 week after seeding the leaves became partially discolored, deformed, and dry, and ultimately about 95% of all the hemp plants died. The surviving hemp produced shorter shoots and much lower amounts of biomass than hemp on unpolluted soil. Studies in pots and hydrocultures indicated that the inhibition of hemp growth is caused by the synergistic action of a low pH and several mobile toxic metals. Hemp on sediment incorporated much higher quantities of heavy metals (especially Zn, Cd, and Ni) than hemp on unpolluted soil, which calls the commercial use of the former seriously in question.     

Isolation and characterization of microsatellite markers in the striped hamster (Cricetulus barabensis)

A total of 14 polymorphic microsatellite markers were isolated and characterized from the striped hamster, Cricetulus barabensis, a widespread rodent pest in northern China. Two to six alleles per locus were detected in 90 individuals from three locations in Shandong Province, China. The observed and expected heterozygosities ranged from 0.21 to 0.78 and from 0.30 to 0.80, respectively. These microsatellite markers provide new tools for investigating the population structure of this species.     

Isolation and characterization of microsatellite markers in Japanese pear (Pyrus pyrifolia Nakai)

Fourteen microsatellite markers were developed from an enriched genomic library of Japanese pear (Pyrus pyrifolia) by selective hybridization. They were characterized using 17 Japanese pear cultivars. The expected heterozygosity and observed heterozygosity ranged from 0.21 to 0.74 and from 0 to 0.88, respectively. Two to 11 alleles were detected per locus, with IPPN09 and IPPN15 judged to amplify multiple loci. IPPN17 was the most informative locus with the lowest probability of identity (0.19). These primers exhibited a high cross‐species transferability between species and genera.     

Biological Control of Infestations of Ditchweed ( Cannabis sativa ) with Fusarium oxysporum f.sp. Cannabis in Kazakhstan

Ditchweed ( Cannabis sativa L.) is widely distributed in the Chu Valley of southeast Kazakhstan and is difficult to control using conventional chemical or mechanical control. Thus, plant pathogens were investigated as potential biocontrol agents. Fusarium oxysporum was isolated from symptomatic C. sativa plants from this area. Twenty-five of the isolated strains of F. oxysporum were pathogenic and host-specific to C. sativa in greenhouse studies. These strains of F. oxysporum f. sp. cannabis were further evaluated as mycoherbicides for control of ditchweed in natural field infestations. Twelve strains showed field control of C. sativa , and the most virulent strain elicited wilt symptoms within 2 weeks of inoculation of field plants. Three different mycoherbicide formulations were evaluated. A birch sawdust formulation was the most effective carrier in the field. Food based formulations were heavily predated by birds, rodents and insects.     

The potential of hemp (Cannabis sativa L.) for sustainable fibre production: a crop physiological appraisal

Hemp (Cannabis sativa L.) fibre can be used as a raw material for paper and textile production. A comprehensive research programme in the Netherlands has concluded that fibre hemp is a potentially profitable crop, having the right profile to fit into sustainable farming systems. This paper presents an appraisal of the crop physiological characteristics and the agronomic potential of hemp. Parameter values of basic crop physiological characteristics such as light interception potential, radiation use efficiency and dry matter partitioning coefficients are given. The effect of crop management decisions such as cultivar choice, sowing date, plant density, and harvest date on the value of these parameters is discussed. A simple crop growth model was used to assess the yield potential of hemp for the climate of the Netherlands. Calculations made for a non-stressed late-flowering hemp crop sown on 15 April and harvested on 15 September give a stem dry matter yield of 17.1 t ha^-1. The effects of advancing or delaying sowing or harvest date on stem yield were calculated. Crop physiological characteristics of hemp are compared to those of kenaf (Hibiscus cannabinus L.). Radiation use efficiency and dry matter partitioning coefficients of the two crops are similar. Base temperatures for development and growth are lower in hemp than in kenaf. In a temperate climate with cool springs, canopy establishment will be more rapid in hemp than in kenaf. Hemp seems an excellent candidate to fill the niche for an annual fibre crop in a temperate climate.     

Isolation of 10 microsatellite markers for mongoose lemurs (Eulemur mongoz)

We present here 10 new microsatellite markers for the mongoose lemur (Eulemur mongoz), nine of which were isolated from E. mongoz and one from Lemur catta. At least 60 individuals were genotyped for each of the 10 loci. Mendelian inheritance at each locus was tested by genotyping five captive families with known pedigrees. All loci were polymorphic and demonstrated simple Mendelian inheritance. The microsatellite markers presented here will be useful for genetic surveys of wild E. mongoz, to gain insights into the genetic background of the captive population, and to aid in the conservation of the species’ genetic diversity.     

Isolation of five polymorphic microsatellite markers in Utetheisa ornatrix (Lepidoptera: Arctiidae)

We isolated five polymorphic microsatellite markers for the arctiid moth Utetheisa ornatrix as part of an effort to investigate the role of inbreeding in the degree of polyandry seen in natural populations of the moth. We tested the primers on 105 individuals collected from scrub habitat in central Florida. These microsatellite loci provide a new research tool for understanding the population structure and reproductive behaviour of U. ornatrix.     

Isolation and characterization of polymorphic microsatellite markers for peacock wrasse (Symphodus tinca)

Eight polymorphic microsatellite loci were isolated and characterized for the peacock wrasse (Symphodus tinca), a labrid fish inhabiting the Mediterranean and Black seas. Characterization of 35 individuals from the western Mediterranean indicated a relatively high allelic diversity (mean = 12.4, range 9–17), and observed heterozygosity ranging from 0.65 to 0.91. We found no evidence of linkage disequilibrium between pairs of loci. Two loci showed significant departure from Hardy–Weinberg equilibrium. These polymorphic markers can be useful in most basic population genetic applications.     

Isolation of polymorphic microsatellite markers for Begonia sutherlandii Hook. f.

Seven polymorphic microsatellite loci have been characterized for investigating population structure in the patchily distributed herb Begonia sutherlandii. Two loci (BSU3 and BSU4) exhibited population specific null alleles; primer redesign and allele sequencing for one of these loci showed two transition mutations in the original primer site. Two loci exhibited imperfect repeat polymorphisms due to single base pair indels in the flanking region (locus BSU6) and in the microsatellite region itself (BSU7). Transversion mutations were also found in the microsatellite region of locus BSU7. The remaining three loci amplified in all individuals tested and appeared to conform to a simple stepwise mutation pattern.