Molecular detection of Penicillium griseofulvum as the coastal pollution indicator.
Molecular methods were carried out to detect Penicillium griseofulvum, a dominant species related to heavy metal pollution, which was screened from marine contaminated sediments. Based on differences in internal transcribed spacer (ITS) sequences of Penicillium genus and specific isoamyl alcohol oxidase (IAO) sequences, species-specific primers AS1/RS4 and IAO1/IAO2 of Penicillium griseofulvum were designed and synthesized which were then employed in optimized PCR systems. The detection sensitivities were compared through ordinary PCR and nested-PCR using two pairs of primers, respectively. Both primer pairs could exclusively amplify destined DNA fragment from contaminated environmental samples in our researches. As for primers AS1/RS4, the detection sensitivity for spores (pure spore DNA) could be 10 fg/μl and 10 spores, respectively, and the detection sensitivity for the sediments was 10² spores/0.25 g sediments. While the detection sensitivity of IAO1/IAO2 primers was lower than that of AS1/RS4. Despite the difference in detection sensitivity, it is feasible that the species-specific primers could be used as probes for the detection of environmental pollution dominant species, Penicillium griseofulvum, since the frequency of occurrence and amount of this strain could preferably indicate the pollution degree.
Shi
Zhengping
Z
Qingdao University of Science & Technology, Qingdao, 266042, China.
Bai
Shumeng
S
Tian
Li
L
Jiang
Huichao
H
Zhang
Jiuming
J
D017931:DNA Primers / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D021903:DNA, Ribosomal Spacer / Q000235:genetics; D004785:Environmental Pollutants / Q000378:metabolism; D004787:Environmental Pollution; D005656:Fungal Proteins / Q000235:genetics; D019015:Geologic Sediments / Q000382:microbiology*; D019216:Metals, Heavy / Q000378:metabolism; D009172:Mycology / Q000379:methods*; D010088:Oxidoreductases / Q000235:genetics; D010407:Penicillium / Q000235:genetics / Q000302:isolation & purification*; D016133:Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity
D017931:DNA Primers; D004271:DNA, Fungal; D021903:DNA, Ribosomal Spacer; D004785:Environmental Pollutants; D005656:Fungal Proteins; D019216:Metals, Heavy; D010088:Oxidoreductases