Detection of somaclonal variation of cotton (Gossypium hirsutum) using cytogenetics, flow cytometry and molecular markers

Abstract

Two protocols of plant regeneration for cotton were adopted in this study, namely, 2, 4-D and kinetin hormone combination and IBA and kinetin hormone combination. Twenty-eight embryogenic cell lines via somatic embryogenesis and 67 regenerated plants from these embryogenic calli were selected and used for random amplified polymorphic DNA (RAPD), simple sequence repeat (SSR), chromosomal number counting, and flow cytometric analysis. The roles of RAPD and SSR markers in detecting somaclonal variation of cotton (Gossypium hirsutum L.) were evaluated. Two cluster analyses were performed to express, in the form of dendrograms, the relationships among the hormone combinations and the genetic variability. Both DNA-based techniques were able to amplify all of the cell clones and regenerated plantlets genomes and relative higher genetic variation could be detected in the culture type with 2, 4-D and kinetin hormone combination. The result suggested that 2, 4-D and kinetin hormone combination could induce relative high somaclonal variation and RAPD and SSR markers are useful in detecting somaclonal variation of regenerated cotton plants via somatic embryogenesis. Chromosome number counting and flow cytometry analysis revealed that the number of chromosomes and ploidy levels were nearly stable in all regenerated plants except two regenerated plantlets (lost 4 and 5 chromosomes, respectively) which meant that cytological changes were not correlated with the frequency of RAPD and SSR polymorphisms. This result also might mean that the cell lines with variation of chromosome numbers were difficult to regenerate plants.

<b>Effects of silicon on the growth and genetic stability of passion fruit

“…However, no reports exist of excess silicon altering the DNA content of plants. In this sense, the flow cytometry technique has gained particular attention because it allows the relative amount of nuclear DNA of plant cells to be estimated rapidly and with high accuracy (Jin et al, 2008;Bairu, Aremu, & Van staden, 2011;Smulders, & Klerk, 2011).…”

“…The flow cytometry technique is useful for distinguishing differences in the DNA content of plants maintained under stressful conditions, such as during micropropagation or when being subjected to chemical treatments. Many economically important species, such as Vitis vinifera (Yang et al, 2008;Prado et al, 2010), Gossypium hirsutum, (Jin et al, 2008), Musa spp. (Msogoya, Grout, & Roberts, 2011;EscobedoGraciaMedrano, Maldonado-Borges, Burgos-Tan, Valadez-González, Ku-Cauich, 2014), Passiflora spp.…”

Morphological and RAPD-marker characterization of Melia volkensii (Grke) in vitro plants regenerated via direct and indirect somatic embryogenesis

“…Epigenetic variations are due to changes in gene expression arising from DNA methylation, histone modifications and small RNA-mediated regulation (Miguel and Marum, 2011;Neelakanda and Wang, 2012). DNA sequence changes can be detected by a variety of molecular markers such as random amplified polymorphic DNA (RAPD) and microsatellites or simple sequence repeats (SSR) (Jin et al, 2008), polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) (Kour et al, 2009) and amplified fragment length polymorphism (AFLP) (Li et al, 2007).…”

Massal Induction of Carica papaya L. ^|^lsquo;Golden^|^rsquo; Somatic Embryos and Somaclone Screening by Flow Cytometry and Cytogenetic Analysis

“…The use of FCM has been an excellent alternative for assessing changes in DNA content and ploidy level (Jin et al 2008). However, this technique cannot detect chromosomal rearrangements.…”