Abstract
Sequence-characterized amplified region (SCAR) markers were used to identify female
plants at an early developmental stage in four different varieties of Cannabis sativa. Using the cetyl trimethylammonium bromide (CTAB) method, DNA was isolated from two-week-old
plants of three drug-type varieties (Terbag W1, Terbag K2, and Terbag MX) and one
fiber-type variety (Terbag Fedora A7) of C. sativa grown under controlled environmental conditions through seeds. Attempts to use MADC2
(male-associated DNA from Cannabis sativa ) primers as a marker to identify the sex of Cannabis sativa plants were successful. Amplification of genomic DNA using MADC2-F and MADC2-R primers
produced two distinct fragments, one with a size of approximately 450 bp for female
plants and one for male plants with a size of approximately 300 bp. After harvesting
the tissues for DNA extraction, plants were subjected to a flowering photoperiod (i.e.,
12-h light cycle), and the appearance of flowers was compared with the DNA analysis.
The results of the molecular analysis were found to be concordant with the appearance
of male or female flowers. The results of this study represent a quick and reliable
technique for the identification of sex in Cannabis plants using SCAR markers at a very early developmental stage.
Key words
Cannabis sativa L. - Cannabaceae -
Cannabis sex - SCAR
References
1 Abel E. Marihuana: the first 2000 years. New York; Plenum Press 1990
2
Hammond C T, Mahlberg P G.
Morphogenesis of capitate glandular hairs of Cannabis sativa (Cannabaceae).
Am J Bot.
1977;
64
1023-1031
3
Mechoulam R, Ben-Shabat A.
From gan-zi-gun-nu to anandamide and 2-arachidonoylglycerol: the ongoing story of
Cannabis .
Nat Prod Rep.
1999;
16
131-143
4 Mechoulam R.
Cannabinoids as therapeutic agents. Mechoulam R Cannabinoids as therapeutic agents. Boca Raton; CRC Press 1986: 1
5
Formukong E A, Evans A T, Evans F.
The medicinal uses of Cannabis and its constitutents.
J Phytother Res.
1989;
3
219-231
6 Grinspoon L, Bakalar J B. Marihuana, the forbidden medicine. New Haven; Yale University
Press 1993
7
Mattes R D, Shaw L M, Eding-Owens J, Egelman K, ElSohly M A.
Bypassing the first pass effect for therapeutic use of cannabinoids.
Pharmacol Biochem Behav.
1993;
44
745-747
8
Mattes R D, Egelman K, Shaw L M, ElSohly M A.
Cannabinoids appetite stimulation.
Pharmacol Biochem Behav.
1994;
49
187
9
Brenneisen R, Egli A, ElSohly M A, Henn V, Spiess Y.
The effect of orally and rectally administered Δ
9 -tetrahydrocannabinol on spasticity. A pilot study with two patients.
Int J Clin Pharmacol Therap.
1996;
34
446
10
Sirikantaramas S, Taura F, Tanaka Y, Ishikawa Y, Morimoto S, Shoyama Y.
Tetrahydrocannabinolic acid synthase, the enzyme controlling marijuana psychoactivity
is secreted into the storage cavity of the glandular trichomes.
Plant Cell Physiol.
2005;
46
1578-1582
11
Sakamoto K, Shimomura K, Komeda Y, Kamada H, Satoh S.
A male-associated DNA sequence in a dioecious plant, Cannabis sativa L.
Plant Cell Physiol.
1995;
36
1549-1554
12
Sakamoto K, Abe T, Matsuyama T, Yoshida S, Ohmido N, Fukui K, Satoh S.
RAPD markers encoding retrotransposable elements are linked to the male sex in Cannabis sativa L.
Genome.
2005;
48
931-936
13
Flachowsky H, Schuhmann E, Weber W E, Peil A.
Application of AFLP for the detection of sex-specific markers in hemp.
Plant Breed.
2001;
120
305-309
14
Törjék O, Bucherna N, Kiss E, Homoki H, Finta-Korpelová Z, Bócsa I, Nagy I, Heszky L E.
Novel male-specific molecular markers (MADC5, MADC6) in hemp.
Euphytica.
2002;
127
209-218
15
Li M, Yang H, Li F, Yang F, Yin G, Gan S.
A male specific SCAR marker in Calamus simplicifolius , a dioecious rattan species endemic to China.
Mol Breed.
2009;
DOI: 10.1007/s11032-009-9349-9
16
Mandolino G, Carboni A, Forapani S, Faeti V, Ranalli P.
Identification of DNA markers linked to the male sex in dioecious hemp (Cannabis sativa L.).
Theor Appl Genet.
1999;
98
86-92
Dr. Natascha Techen, Ph.D
School of Pharmacy National Center for Natural Products Research University of Mississippi
University Ave, P. O. Box 8048
University, MS 38677
United States
Telefon: +1 66 29 15 10 10
Fax: +1 66 29 15 70 62
eMail: ntechen@olemiss.edu