Environmentally induced polymorphisms detected by RAPD analysis of soybean seed DNA

Abstract

Experiments were performed to determine if RAPD profiles developed using total DNA isolated from soybean seed could be affected by the physiological state or the quality of the seed. RAPD profiles were developed using template DNA isolated from a single seed lot of soybean (Glycine max L. cv. Kirby). High quality seeds were used to produce four populations varying in either quality or physiological state: untreated control seed ambient temperature and humidity storage for 12 months, accelerated aging at 41°C and 100% relative humidity for 48 h, and controlled hydration (seed priming). One hundred and eighty-eight primers were used to create separate RAPD profiles from total DNA isolated from each set of seed and from soybean leaf tissue. Sixteen polymorphisms from 14 primers were identified as a result of seed treatments. Six primers showed nine polymorphisms in RAPD profiles of ambient-stored seed DNA, while four and two primers produced polymorphisms in reactions using accelerated aged or primed-seed template DNA, respectively. Two primers showed a polymorphic fragment in vegetative DNA not observed in any of the seed DNA samples. Ten of the observed polymorphisms were due to the appearance of a DNA fragment in response to a specific seed treatment while six were the result of the treatment-induced loss of a DNA fragment. The six polymorphisms resulting from the loss of a major fragment were all due to ambient-temperature seed storage. Results were reproducibly obtained from multiple DNA isolations using three separate DNA isolation procedures involving either multiple seed or a single seed as the template source. Therefore, genetically identical seed can consistently display RAPD polymorphisms as a response to the environmental exposure.

Studies on genetic changes in rye samples (Secale cereale L.) maintained in a seed bank

“…This result is consistent with our previous data, in which we detected some inherited changes in fragments derived from the non-coding genome region in material of low viability (same plant material) [11]. Thus, our results support the hypothesis of Shatter et al [9] that DNA alternations induced by ageing are not randomly dispersed throughout the genome, but are rather concentrated into hot-spots. It seems likely that these hot-spots are present in non-coding DNA regions.…”

“…Methods based on phenotypic observations [4] or protein markers [5][6][7] have been employed to investigate such events. Variation at the DNA level has been investigated using markers such as RAPDs [8,9], AFLPs [10], STSs [11] and microsatellites [12,13]. RAPD and AFLP analyses have indicated that both storage time and the number of regeneration cycles can influence the genetic structure of a population [8,10].…”

Molecular Marker Analysis of Differentially Aged Seeds of Soybean and Safflower

“…Hence, it could be possible in the near future to select SSR primers in the regions flanking the loci of interest, which are known to be associated with longevity behavior such as the efficiency of the radical scavenging system or production of antioxidants and DNA repair enzymes. Shatters et al (1995) observed polymorphisms in the RAPD profiles of DNA from differently aged soybean seed. Similar results were obtained with RAPD (Bednarek et al 1998) and AFLP markers (Chwedorzewska et al 2002a) in long-term stored rye seeds.…”

Stress-Induced Cell Reprogramming. A Role for Global Genome Regulation?

“…There is vast literature available from the past, demonstrating the high variability in repetitive DNA in relation to plant development and stress, including especially tissue culture experiments (see Arnholdt-Schmitt, 1995;Schaefer et al, 2000). Also, environmentally induced DNA polymorphisms were reported for several species (Shatters et al, 1995;Lerner, 1999).…”