Internal Transcribed Spacer Restriction Fragment Length Polymorphism Research Articles

Comparison of ITS RFLP patterns of Gracilaria (Rhodophyceae, Gracilariales) populations from Chile and New Zealand and an examination of interfertility of Chilean morphotypes

Restriction fragment length polymorphism (RFLP) patterns of the internal transcribed spacer (ITS) of the nuclear ribosomal cistron and crossability trials were used to characterize four morphotypes of Gracilaria from Lenga, Isla Santa Maria and Maullin, Chile, and two morphotypes from sites in New Zealand. PCR products from all Chilean morphotypes resulted in a major single band of ca. 1198 bp. ITS-RFLP profiles generated with the restriction enzymes Cla I, Hae III, Pst I, Hha I, Rsa I and Taq I, were identical in all cases. All crosses within, as well as between, morphotypes resulted in cystocarp differentiation, with the production of viable carpospores. Based upon these data, it is concluded that the four morphotypes from Chile correspond to a single species, G. chilensis, and that the ITS-RFLP pattern is a useful marker to predict genetic relatedness at the specific level in Gracilaria. A comparison of the ITS-RFLP patterns of the Chilean morphotypes with the patterns of two samples of G. chilensis from New Zealand revealed that the sample from Scorching Bay, Wellington, fits the Chilean ITS-RFLP patterns. The population from Blockhouse Bay, Auckland, appears to correspond to another species.

Molecular determination of genetic variation in Pisolithus isolates from a defined region in New South Wales, Australia

Sixty‐two isolates of Pisolithus were obtained from a carpophore population collected from a 90‐km2 area of New South Wales, Australia. Random amplified polymorphic DNA (RAPD) analysis was used initially to screen isolates for variation, and restriction fragment length polymorphism (RFLP) analysis of the internal transcribed spacer (ITS) region and ITS sequence analysis were subsequently performed on selected isolates. ITS–RFLP and ITS sequence analysis indicated that two isolates (LJ07 and WM01) from different sites were considerably different from all other isolates, a strict consensus tree generated by bootstrap analysis of sequence data strongly suggesting that isolates LJ07 and WM01 represent a separate Pisolithus species. Although the general morphology of carpophores and basidiospores of the majority of collected isolates was consistent with previous descriptions of P. tinctorius (Pers.) Coker and Couch, basidiospore characteristics suggest that isolates LJO7 and WMO1 might represent a Pisolithus species distinct from P. tinctorius and the previously described Australian species, P. microcarpus (Cooke & Massee) G. Cunn.