Genomic Fingerprinting of <i>Camelina</i>Species Using cTBP as Molecular Marker

Interest on the genus Camelina has recently increased due to the biofuel, or jet fuel, potential of the oil extracted from seeds of the cultivated species Camelina sativa (L.) Crantz. While our knowledge on C. sativa is constantly augmenting, only few studies have been performed on the other species of the genus, which could be a potentially useful material for the genetic improvement of C. sativa. The genus Camelina consists of 11 species, but only six (C. sativa, C. microcarpa, C. alyssum, C. rumelica, C. hispida and C. laxa) could be retrieved from germplasm banks to carry out genomic fingerprinting studies based on the use of the cTBP molecular marker. Each species, with the exception of C. alyssum that is proposed to be a subspecies of C. sativa, shows a distinct cTBP profile resulting from multiple DNA length polymorphisms present in the second intron of the members of the β-tubulin gene family. In contrast to the high level of genetic diversity detected among the six Camelina species, low variability is observed among and within the accessions of the same species, except for C. hispida that is characterized by an intra-accession high number of cTBP polymorphic bands. In addition, cTBP is also able to identify incorrectly classified accessions and provide information on the ploidy level of each species.

Variability in Glucosinolate Content among <i>Camelina</i>Species

Glucosinolate content in Camelina sativa (L.) Crantz (false flax, gold-of-pleasure) and its relatives C. microcarpa, C. alyssum, C. rumelica and C. hispida was investigated. With the exception of C. hispida in which GSL3 was absent, in all remaining species, three characteristic glucosinolates (GSL1, GSL2 and GSL3) were identified. Camelina genotypes of spring type (C. sativa CAM134, C. alyssum CAM21) showed a typical pattern of glucosenolates with GSL1 > GSL3. GSL1 was present in traces in C. microcarpa and at low levels in C. rumelica and C. alyssum subsp. alyssum. In C. hispida, the GSL1 content was greater than GSL2 and, only in this specie, GSL2 represented less than 50% of total glucosinolates. These differences in the glucosinolate pattern among Camelina species could be exploited to reduce the total content of glucosinolates in C. sativa.

<i>In Silico</i>Exploration of <i>Cannabis sativa</i>L. Genome for Simple Sequence Repeats (SSRs)

Simple sequence repeat (SSR) or microsatellite markers, are a valuable tool for several purposes such as evaluation of genetic diversity, fingerprinting, marker assisted selection, and breeding. Recent developments in sequencing technologies and bioinformatics analyses provide new opportunity to produce a high number of less costly SSRs. Here, we used for the first time a wholegenome shotgun sequencing of the nuclear genome and transcriptome of hemp to develop microsatellite markers for C. sativa L. (hemp). Hemp is an ancient crop that is widely cultivated as a source of fiber, seeds and medicine. The analysis using the MISA program revealed a total of 407,491 SSRs (from mono-nucleotide to deca-nucleotide) in the hemp genome and 15,655 SSRs in the transcriptome. Analysis of the frequency and distribution of SSRs showed that the mono-nucleotide repeats were the most abundant (55.4%) in the genome whereas the tri-nucleotide motifs (30.4%) resulted highly predominant in the transcriptome. Poly A/T was predominant over poly G/C in both genome and transcriptome sequences. Among the tri-nucleotide repeats AAG/CTT (34.5%) resulted the most abundant in the transcriptome. Repeats larger than tri-nucleotide were also observed in the hemp genome and transcriptome. Dinucleotide and tri-nucleotide repeat expansion of 8605 and 1401 times iteration were observed however, other SSR expansion more than 387 times repetition was not found. Primers were designed for amplification of few long microsatellite sequences which could be used to identify polymorphism and to study genetic diversity among hemp cultivars.