The polyene antibiotics, including nystatin, pimaricin, amphotericin, and candicidin, comprise a family of very valuable antifungal polyketide compounds, and they are typically produced by soil actinomycetes. Previously, using a polyene cytochrome P450 hydroxylasespeciWc genome screening strategy, Pseudonocardia autotrophica KCTC9441 was determined to contain genes potentially encoding polyene biosynthesis. Here, sequence information of an approximately 125.7-kb contiguous DNA region in Wve overlapping cosmids isolated from the P. autotrophica KCTC9441 genomic library revealed a total of 23 open reading frames, which are presumably involved in the biosynthesis of a nystatin-like compound tentatively named NPP. The deduced roles for six multimodular polyketide synthase (PKS) catalytic domains were found to be highly homologous to those of previously identiWed nystatin biosynthetic genes. Low NPP productivity suggests that the functionally clustered NPP biosynthetic pathway genes are tightly regulated in P. autotrophica. Disruption of a NPP PKS gene completely abolished both NPP biosynthesis and antifungal activity against Candida albicans, suggesting that polyene-speciWc genome screening may constitute an eYcient method for isolation of potentially valuable previously identiWed polyene genes and compounds from various rare actinomycetes widespread in nature.
Gametogenesis, mating behaviour and spawning of Octopus ocellatus were investigated by histological study. This species is dioecious, and showed a protandry phenomenon. Ooogenesis (in females) and spermatogenesis (in males) can be classified into 3 stages, respectively. O. ocellatus copulates in one of two ways: a male may leap upon a female, mounting her mantle, or a male may sit near the female and extend the hectocotylized third right arm toward her. Spawning occurred between April and June in females, and between March and May in males of O. ocellatus. The spawning period was once a year and the peak took place between May and June. A number of flatened follicle cells, which were attached to an oocyte, were involved in vitellogenesis in the cytoplasm of the vitellogenic oocyte (maturing oocyte), and formation of chorion membrane (secondary egg membrane) of the ovarian eggs. Fecundity per female closely related to GSI was 294-660 eggs (average, 429 eggs). The díameters of the ovarían eggs surrounded by chorion membrane were approximately ín the range of 10.10-2.50 mm. Each ovarian egg laid by a female was connected to an egg string. Each egg string was 1-5.5 cm (average 3.6 cm). The total number of eggs laid by a female of this species ranged 218-314, the egg sizes were independent to the size of female adult. this species has a life mode showing some special reproductive characteristics of an annual semelparity as shown in Octopodidae species because we have never seen a female spawning a second time.
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