"We treated our Arabidopsis plants with a mutagen to try to trigger mutations that would increase oil accumulation," Xu said, using the fluorescence technique to identify strains with more and/or bigger green dots. Ironically, they made their discovery about sterol in an Arabidopsis strain that accumulated almost no oil. "The main purpose of the current work was to figure out which genetic modification caused this dramatic drop in oil accumulation," Xu said. "We thought tracking down this gene might give us some new genes/proteins that are important in lipid droplet formation or accumulation." At the microscopic scale, scientists know that lipid droplets form in the "endoplasmic reticulum," or ER, of cells. That's an internal network of membranes within cells (not the membrane surrounding the cell) that acts like a sort of factory -- assembling and packaging various materials such as proteins and lipids. Lipid storage droplets form when oil begins to accumulate between the two layers of the ER membrane, but only in certain regions of the ER. Eventually, when enough oil is present, the little membrane bits pinch off leaving the oil encapsulated in self-contained compartments. As the study at Brookhaven shows, studying a plant that doesn't accumulate these lipid droplets can offer clues to the biochemical factors that drive the process -- and what's unique about the particular ER domains where it occurs. To figure out which mutation triggered the dramatic drop in oil accumulation, the Brookhaven team used a technique known as positional cloning -- a way of searching every region of chromosomes to pinpoint a particular gene that is responsible for a characteristic of interest. The technique narrowed the search to a specific region in one of the plant's chromosomes. "This region still contains hundreds of candidate genes," Xu said. After using whole-genome sequencing to search for any mutation in this region, the team identified a gene they suspected was involved. The gene codes for an enzyme responsible for one biochemical step in the multi-step synthesis of sterol, a molecule related to cholesterol found in ER and other cellular membranes. By selectively "knocking out" the normal (unmutated) version of this gene, the scientists were able to duplicate the effect of the mutation. That is, plants with the gene knocked out accumulated no lipid droplets. In addition, adding back the unmutated gene restored oil droplet accumulation. "This experiment provided clear evidence that sterol plays an essential role in forming oil droplets," Xu said. But the scientists went further. They also studied what would happen if they mutated genes for enzymes "upstream" from this particular enzyme in the multi-step sterol synthesis pathway. And they measured sterol levels in these mutants. The detailed studies allowed them to zero in on the specific type of sterol that, when deficient, results in low oil accumulation.https://www.sciencedaily.com/releases/2021/09/210922090838.htm
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Just $5 a month. Under Prime Minister David Lange, New Zealand’s fourth Labor government had refused entry to the USS Buchanan warship in 1985. The United States had dispatched the ship to New Zealand as a test of the new Labor government’s nuclear-free policy. U.S. President Ronald Reagan’s Republican administration refused to confirm or deny the ship’s nuclear capability. New Zealand’s subsequent decision to refuse the ship permission to enter unleashed the United States’ fury. As punishment, New Zealand was effectively kicked out of the ANZUS defense alliance – of which it had been a member since 1951 – and was stripped of its status as a U.S. ally. George Shultz, Reagan’s secretary of state, made clear his view of the new U.S. position toward New Zealand: “The time has come to part. We part as friends – but we part.” There had been some improvement to the NZ-U.S.https://thediplomat.com/2021/09/how-9-11-changed-new-zealands-foreign-policy/