Induction of embryo development and fixation of partial interspecific lines after pollination of F1 cotton interspecific hybrids (Gossypium barbadense × Gossypium hirsutum) with pollen from Hibiscus cannabinus

Abstract

The possibility of inducing embryo development after pollination of F1 interspecific cotton hybrids (Gossypium barbadense × Gossypium hirsutum) and their reciprocals with pollen from Hibiscus cannabinus was investigated. For this, flowers of F1 plants from 4 G. barbadense × G. hirsutum interspecific hybrids (B403 × Acala Sindos, Carnak × 4S, B403 × Coker 310, and Carnak × Acala Sindos) and their reciprocals grown in the field were pollinated with pollen from Hibiscus cannabinus. From the 443 pollinated flowers, 276 were left on the plant to grow naturally, and 167 were collected 5 days after pollination. Young ovules from the collected buds were cultured in vitro for embryo development. It was observed that, from the buds left to grow naturally on the mother plant, 21 bolls reached maturity. The mature bolls originated only from the 4 G. barbadense × G. hirsutum hybrids and contained 82 mature seeds. Finally, 38 plants (Pa0) were produced. From the in-ovule culture method, 10 young embryos were isolated from both G. barbadense × G. hirsutum and G. hirsutum × G. barbadense hybrids and finally 3 plants were produced. The plants produced from both approaches originated only from the G. barbadense × G. hirsutum hybrids. These plants exhibited morphological traits from both cotton species and they were partially fertile. No signs of H. cannabinus morphological traits were observed in the plants produced. Root-tip chromosome counts revealed that chromosome number among cells of the Pa0 plants ranged from 27 to 42 and the difference in chromosome number observed among cells of the same plant ranged from 1 to 3. The chromosome number, however, was increased progressively from generation to generation and in Pa3 it ranged from 46 to 52. Plants with 52 chromosomes were identified even from the Pa1 generation. In addition, flow cytometric analysis indicated that the parental plants had a similar DNA profile to the F1 and F2 interspecific hybrids but a different one from the Pa0 plants. Thus, alien pollination of cotton flowers from interspecific (G. barbadense × G. hirsutum and reciprocals) hybrids with pollen from H. cannabinus most likely induced parthenogenetic (Pa) egg cell development which, after a progressive chromosome increase, produced fully fertile plants with most of the cells at the tetraploid or near-tetraploid level. It was concluded that a combination of the in situ boll development with an optimised in vitro ovule culture technique could establish the ‘cannabinus method’ in cotton, as a method for the production of genotype-independent partial interspecific lines.

In vitro gynogenesis in cotton (Gossypium sp.)

“…However, in the present study, we did not observe any significant differences among genotypes. We chose to use only G. barbadense cultivars as the female parent, because it has been reported that ovules of G. hirsutum 9 G. barbadense cotton hybrids do not react to in vitro culture (Mavromatis et al 2005).…”

“…In cotton haploid production, species within the Malvaceae could be used as potential inducers of gynogenesis. H. cannabinus has been reported to cause in situ parthenogenesis in cotton (Zhou et al 1991;Mavromatis et al 2005). There is a natural incompatibility system that prevents hybridization and introgression with cotton (Kantartzi and Roupakias 2008a).…”

Production of aneuploids of the cotton hybrid G. barbadense × G. hirsutum L. via intergeneric pollination with Abelmoschus esculentus

“…Since all attempts for the creation of pure lines with the use of cotton monosomics (Galanopoulou-Sendouca and Roupakias 1999), haploid production by in-vitro anther and microspore culture (Bajaj and Gill 1996; Goodin and Trolinder 1990;Turaev and Shamina 1986;Mavromatis and Roupakias 1994) or unfertilized ovules (Mavromatis and Roupakias 1995) have been unsuccessful, the only alternative remaining for the production of dihaploids in relatively large numbers was pollination with alien pollen. Accordingly, Mavromatis et al (2005) attempted to pollinate F 1 cotton hybrids (G. barbadense · G. hirsutum) with pollen from Hibiscus cannabinus. This work resulted mixoploid fertile plants in which somatic chromosome numbers ranged from 27 to 42.…”

“…The next step is to investigate the possibility of obtaining plants in advanced generations with increased and finally stabilized number of chromosomes at the level of 52, as occurred in crosses of the interspecific hybrid with H. cannabinus (Mavromatis et al 2005). If euploidy is not obtained we will try to exploit the system of alien pollination for the development of aneuploid plants without the use of radiation.…”

“…Zhou et al (1991Zhou et al ( , 1992Zhou et al ( , 2003 reported on parthenogenetic seed set (which reached maturity on the mother plant) at the rate of 41 seeds per 100 pollinated flowers when G. hirsutum F 1 intraspecific hybrid plants were pollinated with pollen from Hibiscus cannabinus (2n = 32). Likewise, Mavromatis et al (2005) reported the development of partial interspecific cotton plants via alien pollination of F 1 interspecific hybrids of G. barbadense · G. hirsutum with pollen from H. cannabinus L. These plants had combined characteristics from both cotton species, were fertile and their chromosome number was increased in three generations back to 52, the normal chromosome number of the tetraploid cottons. The idea of the creation of partial interspecific lines in cotton by pollination of F 1 interspecific hybrids with pollen derived from Abelmoschus esculentus (L.) Moench (2n = 62-65), is very appealing, because in Greece the latter is an important vegetable crop grown as an annual crop during the summer, and its flowering season coincides with cotton.…”