In the present study, genetic polymorphism and diversity in unicellular clones of Chlorella vulgaris Beijerinck and Chlorella pyrenoidosa Chick were studied with Inter Simple Sequence Repeats PCR (ISSR PCR). Samples including four clones of C. vulgaris and three clones of C. pyrenoidosa were purified by single-clone-choice method. For four C. vulgaris unicellular clones, the total number of the bands scored for 18 primers was 298; and the number of the polymorphic bands was 118, of which 39.6% were polymorphic. The size of PCR products ranged from 200 to 2 500 bp. The total number of bands scored for 18 primers, the number of polymorphic bands and the percentage of three C. pyrenoidosa unicellular clones was 194.83 and 30.8%, respectively. POPGENE analysis show that the average Nei genetic diversity (h^*) and Shannon index of diversity (I^*) in the four C. vulgaris unicellular clones was 0.2181 and 0.3208, respectively, which is slightly higher than those of the three C. pyrenoidosa unicellular clones (0.190 3 and 0.274 8), which agreed with the percentage of polymorphic bands in the mixed samples of the two species. The results suggest that ISSR is a useful method to Chlorella for intra-species genetic analysis.
Vertical polyamide gel electrophoresis was used to investigate isozyme polymorphisms among different isolates (including wild and cultivated) of Porphyra katadai, Porphyra oligospermatangia, Porphyra yezoensis, Porphyra haitanensis, and a hybridize species (Porphyra yezoensis x Porphyra haitanensis) sampled from China. Whereafter, the analyses of probable minimum loci numbers, observed alleles sum, genetic diversity, and unweighted pair-group arithmetic average (UPGMA) cluster were carded out. After initial activity and resolution testing of bands of 23 enzymes, 6 of them (MDH, ME, LDH, GDH, IDH and G-6-PDH) were proved to be appropriate for analysis of the full sample set. The probable minimum numbers of loci and alleles analyses showed that the five species of Porphyra had an extraordinary consistent result in ME loci and alleles. However, P. katadai and P. oligospermatangia differed from other three species of Porphyra in LDH and GDH loci and alleles. P. katadai was independent in the analyses of MDH and P. oligosperTnatangia and P. haitanensis differed from other three species in IDH analyses. Moreover, P. yezoensis and P. haitanensis were apart from other three species in G-6-PDH analysis. Taking one with another, P. katadai was relatively separated in the probable minimum numbers of loci and alleles analyses. The results indicated that the genetic variation among the five Porphyra species was limited with a genetic identity of 0.7550. The hybridize species (P. yezoensis x P. haitanensis) seemed to be high homologue with P. oligospermatangia, unexpectedly got relatively lower average genetic identities with both P. yezoensis and P. haitanensis. The 4 strains of P. yezoensis were relatively divergent with an average genetic identity of 0.7428, and P. katadai presented the most differentiated, compared with other species, which consistented with the result summarized in the probable minimum numbers of loci and alleles analyses.
Fe2+ acted as an accessorial factor for many cellular enzymatic reactions is very important for seaweed growth and development, but the Fe2+ requirement in nori had not been seen. Porphyra yezoensis cells were separated enzymatically and cultured in a series of sterilized seawater media containing various concentra- tions of Fe2+. The growth development and cell were investigated in this work. Through this experiment, two biologically-meant concentration scales were found, one is low concentrations, 12.1–102.1 μg/L, 10–100 times than that in seawater, favoring the development of isolated cells of Porphyra and the other was high concentra- tions, more than 10mg/L inhibiting the cell growth, leading to the deformity and shrinkage of the cells. At the concentration of 50 mg/L, the cells stopped growing and died eventually.