Coniothyrium minitans is an important sclerotial and hyphal parasite of the plant pathogen Sclerotinia sclerotiorum. Previously, a conidiation-deficient mutant, ZS-1N22225, was screened from a T-DNA insertional library of C. minitans. CmVps39, a homologue of Vam6p/Vps39p that plays a critical role in vacuolar morphogenesis in yeast, was disrupted by a T-DNA insertion in this mutant. CmVps39 is composed of 1071 amino acids with an amino-terminal citron homology domain and a central clathrin homology domain, as observed for other Vam6p/Vps39p family proteins. Abnormal fragmented vacuoles were observed in ΔCmVps39 under light microscopy and transmission electron microscopy, and ΔCmVps39 showed impairment in autophagy. ΔCmVps39 also exhibited significantly reduced hyphal development, poor conidiation and decreased sclerotial mycoparasitism. In addition, deletion of CmVps39 affected osmotic adaptation, pH homeostasis and cell wall integrity. Taken together, our results suggest that CmVps39 has an essential function in vacuolar morphology, autophagy, fungal development and mycoparasitism in C. minitans.
Seed dormancy is crucial for plant survival and prevents seed germination out of season. However, little is known about the regulatory mechanism of morphophysiological seed dormancy. Ginkgo biloba L. is one of the most ancient gymnosperms, and the completion of seed germination in this species requires cold and moist stratification. Here, we observed that at the mature seed stage, the embryo was not fully developed in G. biloba seeds. During dormancy stages, the length and weight of the embryo significantly increased, and nutrients accumulated in cotyledons. We further found that abscisic acid (ABA), gibberellic acid (GA), cytokinin and ethylene were integrated in the seed dormancy induction, maintenance and release processes, and GA biosynthesis and signaling transduction specifically act on dormancy release. Combining mRNA and miRNA analyses, we demonstrated that miRNA156 is involved in the regulation of morphophysiological dormancy. Our analyses revealed that G. biloba seed dormancy belongs to the ancestral morphophysiological dormancy type, which is not only regulated by the balance of ABA/GA, but also by other hormones associated with embryo morphological development, as well as genes related to embryo differentiation and development. These findings helped with elucidating the comprehensive regulatory network of morphophysiological dormancy in tree seeds.
An automatic nucleic acid extraction device was developed by combining magnetic separation technology and automatic control technology. It was focused on kinematic positioning of the robotic arm and achieved separation and purification of nucleic acids through the magnet transferred technology. This paper presents a device with mechanical arm positioning system, magnetic separation system, and temperature control system. A touch screen was applied as the man-machine interaction. Also, it was featured with the functions for setting parameters and ultraviolet sterilization. Experimental results showed that the high automation device was stable and reliable with excellent extraction effect.
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