Molecular and cytogenetic description of somatic hybrids between Gentiana cruciata L. and G. tibetica King

Abstract

Somatic hybridization provides an opportunity to create cells with new genetic constitution. Here, the interspecific somatic hybrid plants regenerated in vitro following fusion of cell suspension-derived protoplasts of tetraploid Cross Gentian (Gentiana cruciata L., 2n = 52) with protoplasts released from mesophyll tissue of another tetraploid species, Tibetan Gentian (G. tibetica King, 2n = 52), were studied. According to the results of genome analyses with AFLP, ISSR, and CAPS markers, all somatic hybrids were genetically closer to "suspension" fusion partner G. cruciata than to "mesophyll" partner G. tibetica, but they got G. tibetica chloroplasts. Chromosome counting revealed little variation in the number of chromosomes in hybrid's cells (2n = 88 or 2n = 90), although all plants possessed similar nuclear DNA content which remained stable even after 2 years of in vitro culture. Fluorescence in situ hybridization (FISH) showed that hybrids possessed 4 to 7 chromosomes bearing 5S rDNA sites and 6 or 7 chromosomes with 35S rDNA sites. A part of FISH signals was smaller than those observed in the parental species, which could indicate the loss of rDNA sequences. Genomic in situ hybridization (GISH) showed the predominance of the number of G. cruciata chromosomes over chromosomes of G. tibetica. However, a significant level of cross-hybridization was observed for about one-third of hybrid chromosomes, indicating a high degree of homeology between the genomes of G. cruciata and G. tibetica.

Genomic Instability in Somatic Hybridization between Poncirus and Citrus Species Aiming to Create New Rootstocks

“…These losses of chromosomes of the leaf parent are evidence for genomic instability after protoplast fusion during the plant regeneration process. Such kinds of chromosome elimination and genomic instabilities after protoplast fusion have been widely described in other plants such as Brassicaceae species [44], Nicotiana + Atropa [47], Oryza sativa subsp japonica, Oryza sativa subsp indica [54], Gentiana cruciate, and G. tibetica [53]. These instabilities are generally associated with the in vitro process of obtaining protoplast and regenerating plants, but also with "genome shock" [108] in cases of wide hybridization.…”

“…Segregation of organelles [44][45][46] and chromosome instability after protoplast fusion have also been described in other plant species. These instabilities involved whole and partial chromosome elimination or recombination [47][48][49][50], as well as unexpected ploidy levels [51][52][53]. They were interpreted as the consequence of a genomic shock resulting from the merger of two highly differentiated nuclear genomes, leading to rapid and extensive alterations at the genetic and epigenetic levels [53][54][55][56].…”

“…These instabilities involved whole and partial chromosome elimination or recombination [47][48][49][50], as well as unexpected ploidy levels [51][52][53]. They were interpreted as the consequence of a genomic shock resulting from the merger of two highly differentiated nuclear genomes, leading to rapid and extensive alterations at the genetic and epigenetic levels [53][54][55][56]. In order to improve the efficiency of breeding programs based on somatic hybridization at a wide taxonomic level, it is therefore essential to perform in-depth molecular characterizations of the regenerated plants and, particularly, to analyze the inheritance of the genomic regions implied in the expression of selected traits.…”

Determination of Carbohydrates Content in Gentiana Cruciata L. By Gc/MS Method

“…Gentiana cruciata L. owing to its blue flowers and easy cultivation, is popular in gardening [21]. Its aboveground and underground parts are the source of secoiridoid glycosides such as swertiamarine, sweroside and gentiopicroside [22][23][24][25][26]. There is established the pharmacological usage of Gentiana cruciata L., such as antimicrobial, anticholinesterase, hepatoprotective, antigenotoxic and antioxidant [27].…”

Determination of amino acids of some plants from Gentianaceae family

“…They include Gentiana cruciata L. But modern biotechnological methods offer alternative approaches to traditional cultivation methods, leading to rapid micropropagation of Gentiana cruciata L (Budniak et al 2021a). The Gentiana cruciata herb and roots are a source of secoiridoid glycosides, such as sweroside, swertiamarine and gentiopicroside (Szucs et al 2002;Tomiczak 2020). Some pharmacological applications have been identified in Gentiana cruciata L., namely anticholinesterase, antigenotoxic, antioxidant and antimicrobial activities (Olennikov et al 2019).…”