Genetic variation detectable with molecular markers among unadapted germ-plasm resources of cultivated peanut and related wild species

Genome ◽  
1991 ◽  
Vol 34 (6) ◽  
pp. 1013-1020 ◽  
Author(s):  
Tracy M. Halward ◽  
H. Thomas Stalker ◽  
Elizabeth A. Larue ◽  
Gary Kochert
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Wild Species ◽  
Germ Plasm ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reaction ◽  
Cultivated Peanut ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Peanut germ plasm consists of the cultivated allotetraploid species Arachis hypogaea L. and a large number of wild species, which are nearly all diploids. Our previous work indicated a very low level of genetic variability in American cultivars, as assayed by restriction fragment length polymorphism (RFLP) analysis. Since American cultivars might represent a narrow genetic base, we expanded our study to include unadapted germ-plasm lines from the various South American centers of origin, Africa, and China, where considerable morphological and physiological variability has been reported to exist. Wild species of section Arachis were included in the evaluations since they show a high degree of variation when assayed by RFLPs. Three methods were used to assay for RFLP variation: (i) conventional RFLP analysis using random genomic clones from peanut and cDNA clones from peanut and alfalfa (Medicago sativa); (ii) polymerase chain reaction (PCR) amplification of random primer sequences; (iii) four-cutter analysis of PCR-amplified fragments. In all cases a very low level of variability was found in cultivated peanut, while abundant variability was present among wild diploid species. The results are discussed in terms of peanut evolution and significance to peanut breeding.Key words: polymerase chain reaction, Arachis hypogaea, restriction fragment length polymorphism.

Genetic analysis of nuclear DNA restriction fragment patterns

Genome ◽  
1991 ◽  
Vol 34 (5) ◽  
pp. 693-703 ◽  
Author(s):  
Elizabeth M. Gillet
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Genetic Analysis ◽  
Restriction Fragment ◽  
Length Polymorphism ◽  
Fragment Length Polymorphism ◽  
Rflp Analysis ◽  
Chain Reaction ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Restriction fragment length polymorphism (RFLP) analysis in the broad sense is the analysis of differences in restriction fragment pattern produced by defined target segments within or between cell compartments, cell types, etc., in a single individual or in different individuals. Thus both molecular hybridization and DNA amplification by two-primer extension using the polymerase chain reaction can define target segments for RFLP analysis. The two techniques are outlined with special consideration of characteristics important for genetic analysis. The mode of inheritance of restriction fragment patterns as a prerequisite for their use as genetic markers in inheritance studies is explained, leading to criticism of common usage. The importance of internal restriction sites for the determination of allelic variation is stressed. It is shown that, if target segments are under the control of a single nuclear diploid restriction fragment locus, then complete reconstruction of all parental target segments requires controlled crosses between individuals of like restriction fragment pattern.Key words: genetic analysis, inheritance, restriction fragment length polymorphism, controlled cross, polymerase chain reaction.

scholarly journals Molecular Evidence for Progenitorial Species of Garden Petunias Using Polymerase Chain Reaction—Restriction Fragment Length Polymorphism Analysis of the Chs-j Gene

HortScience ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 300-303 ◽  
Author(s):  
Xinwang Zhang ◽  
Ikuo Nakamura ◽  
Masahiro Mii
Keyword(s):  
Polymerase Chain Reaction ◽  
Restriction Fragment Length Polymorphism ◽  
Length Polymorphism ◽  
Fragment Length ◽  
Wild Species ◽  
Fragment Length Polymorphism ◽  
Chain Reaction ◽  
Pcr Rflp ◽  
Polymerase Chain ◽  
Restriction Fragment Length

Wild species and cultivars of Petunia were subjected to analysis for clarifying the historical progenitors of garden petunias (Petunia hybrida) using polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) of Chalcone synthase J (Chs-J) gene. The PCR products for Chs-J intron with adjacent part of exons digested with RsaI enzyme revealed that P. integrifolia and P. inflata (P. integrifolia complex), both with purple flowers, had one large-sized band [650 base pairs (bp)], whereas P. axillaris (P. axillaris complex), a white-flowered species, showed two smaller-sized bands (200 or 280 bp and 350 bp). In P. axillaris, two different band patterns were found among the three subspecies: 200 and 350 bp for P. axillaris subsp. axillaris and 280 and 350 bp for P. axillaris subsp. parodii and subsp. subandina. The 200-bp band was revealed to be specific to P. axillaris subsp. axillaris. P. hybrida cultivars showed four different band patterns, each of which consisted of two to three of the four bands (200, 280, 350, and 650 bp) detected in the wild taxa examined. These results indicate that the wild species analyzed here might partially have contributed to the Chs-J gene of garden petunias analyzed and demonstrate the use of PCR-RFLP in establishing relationships among closely related species and cultivars of Petunia. The puzzling problem related to the possible contribution of more than one subspecies of P. axillaris is discussed.

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