Danielle Garceau
Year: 2014
Faculty Advisor: Klaas Van Wijk

Analysis of Arabidopsis triple mutant of Plastoglobule-localized ABC1 Kinases 1/3/6

Project  Summary

In chloroplasts, plastoglobules (PGs) are lipoprotein particles contiguous with the outer lipid leaflet of the thylakoid membrane that act as sites of passive lipid deposition and possibly protein catabolite removal during thylakoid turnover. The presence of PG-localized enzymes suggests that PGs may also act as sites of chloroplast metabolism. The PG-localized enzymes ABC1 kinases (ABC1Ks) may be crucial to maintaining photosynthetic capacity during high light stress by regulating the recycling of tocopherols that act as antioxidants to protect the photosystems. The double mutant abc1k1abc1k3(k1k3) has been found to exhibit enhanced light sensitivity under high light stress. ABC1K1 and ABC1K3 likely form a complex and may stabilize ABC1K5 and ABC1K6 which are thought to function similarly in plastid carotenoid metabolism. To determine if ABC1K6 interacts with or is stabilized by ABC1K1 and ABC1K3, our study aimed to isolate and analyze the triple mutant k1k3k6. F2 progeny from k1k3 X k6were genotyped by PCR amplification to isolate knockout mutants. Genotypes were confirmed through RT-PCR and Western Blotting. Chlorophyll fluorescence as well as chlorophyll and carotenoid levels were measured at normal light conditions.    

My Experience

It was wonderful to experience BTI during this summer internship, a place with so many fellow plant enthusiasts.  Together we learned about current research in plant science at weekly seminars and participated in weekly bioinformatics workshops.  I gained further experience and confidence in working independently and efficiently with molecular techniques such as PCR, and broadened my laboratory skills to include protein analysis techniques such as Western Blotting. Working in a Cornell lab exposed me to what life as a graduate student would be like and has affirmed my plan of attaining a PhD and career in Plant Biology.