Proceedings of the XLV Italian Society of Agricultural Genetics - SIGA Annual Congress

Salsomaggiore Terme, Italy - 26/29 September, 2001

ISBN 88-900622-1-5

Poster Abstract

RAPD POLYMORPHISM IN TOBACCO

DEL PIANO L., ACANFORA F., ABET M., SORRENTINO C., COZZOLINO E., CUCINIELLO A., BARBATO L.

Istituto Sperimentale per il Tabacco, Via P. Vitiello n.66, Scafati (SA) Italy

istgenetica@uniserv.uniplan.it

tobacco, RAPD

The molecular marker techniques have become fundamental tool for plant scientist as they can provide a relatively unbiased method of quantifying genetic diversity in plants. The technique known as RAPD (Random amplified Polymorphic DNA) based on PCR amplification of random DNA segments from single arbritary primers, does not require prior DNA sequence information which is instead necessary for most of the other PCR assays.

In tobacco the RAPD technique has been used mainly to identify markers linked to genes for resistance to pathogens, while little information is available about the application of this technique to reveal genetic variability in Nicotiana tabacum.

In this work we have applied RAPD technique to compare random amplification of DNA from the following 12 lines of tobacco, belonging to the collection of the Istituto Sperimentale per il Tabacco: Burley IST G14, Burley IST 23, Bright IST G189, Bright IST MS82, Maryland Md 609, Kentucky IST T720, Havanna IST T762, Xanthi IST X2H, Erzegovina IST E8A, Perustiza IST Pr61, Samsun Katerini 2/5, Samsun T1116.

In order to select primers producing positive and repetable patterns, 180 arbitrary primers were checked, utilizing tobacco DNA from Samsun T1116 line: 12 primers failed to amplify DNA, while 168 revealed from 1 to 11 amplification products per primer, with a size ranging from 0.2 to 2.6 Kbp. Among them, 30 primers were selected to perform RAPD analysis on the 12 tobacco lines. The amplification experiments with 22 primers showed no polymorphic bands while with the others 8 primers showed one or two polymorphic bands only. These results indicate that a low degree of polymorphism is revealed in tobacco by RAPD technique.