title: 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.
authors.affiliation: National Key Laboratory of Crop Genetic Improvement, Huazhong Agricultural University, 430070, Wuhan, Hubei, China. jsx@mail.hzau.edu.cn
authors.forename: Shuangxia
authors.initials: S
authors.lastname: Jin
authors.forename: Ramesh
authors.initials: R
authors.lastname: Mushke
authors.forename: Huaguo
authors.initials: H
authors.lastname: Zhu
authors.forename: Lili
authors.initials: L
authors.lastname: Tu
authors.forename: Zhongxu
authors.initials: Z
authors.lastname: Lin
authors.forename: Yanxin
authors.initials: Y
authors.lastname: Zhang
authors.forename: Xianlong
authors.initials: X
authors.lastname: Zhang
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods; D032461:Chromosomes, Plant / Q000235:genetics; D016000:Cluster Analysis; D003582:Cytogenetics / Q000379:methods*; D005434:Flow Cytometry / Q000379:methods*; D014644:Genetic Variation; D018745:Genome, Plant; D003368:Gossypium / Q000196:embryology / Q000235:genetics* / Q000502:physiology; D007701:Kinetin / Q000494:pharmacology; D018895:Microsatellite Repeats / Q000235:genetics*; D010937:Plant Growth Regulators / Q000494:pharmacology; D019105:Random Amplified Polymorphic DNA Technique; D012038:Regeneration / Q000187:drug effects / Q000235:genetics / Q000502:physiology
chemical_list: D010937:Plant Growth Regulators; D007701:Kinetin