J Integr Plant Biol. ›› 2007, Vol. 49 ›› Issue (3): -.DOI: 10.1111/j.1744-7909.2007.00402.x

• Stress & Phytochemistry •    

Assessment of Genetic Stability Among In Vitro Plants of Arachis retusa Using RAPD and AFLP Markers for Germplasm Preservation

Rachel Fatima Gagliardi, Luiz Ricardo Hanai, Georgia Pacheco, Carlos Alberto Oliveira, Leonardo Alves Carneiro, Jose Francisco Montenegro Valls, Elisabeth Mansur and Maria Lucia Carneiro Vieira   

Abstract: Arachis retusa Krapov. et W. C. Gregory et Valls is endemic in the West-central region of Brazil, occurring in areas endangered by human actions. The establishment of in vitro preservation methods for wild species of Arachis is an alternative to seed banks for germplasm storage, multiplication and distribution. The risk of genetic changes induced by tissue culture and the monitoring of the genetic stability of the biological material before, during and after storage must be considered in the context of conservation. Random amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP) fingerprinting were used to evaluate the genetic stability of in vitro plants originated from cotyledons and embryo axes of A. retusa. Cotyledons originated shoots through direct organogenesis and embryo axes displayed multishoot formation induced by 110 mmol/L and 8.8 mmol/L BAP, respectively. Ninety genomic regions (loci) generated from RAPD and 372 from AFLP analyses were evaluated. All amplified fragments detected by both techniques in plants derived from the two explant types were monomorphic. The results indicate that the recovered shoots are genetically stable at the assessed genomic regions.

Key words: amplified fragment length polymorphism, Arachis retusa, in vitro preservation, micropropagation, random amplified polymorphic DNA, somaclonal variation.

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