Mg 2+ uptake QTLs Hannah Itell and Eric Sawyer. Biological roles for Mg 2+ ●chlorophyll ●common cofactor (RNA pol, kinases, ATPases, …) ●most Mg 2+ is.

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Mg 2+ uptake QTLs Hannah Itell and Eric Sawyer

Biological roles for Mg 2+ ●chlorophyll ●common cofactor (RNA pol, kinases, ATPases, …) ●most Mg 2+ is not free in cytoplasm (ATP/nucleotide-chelated or in organelles) < 2 mM chlorophyll a

Biological roles for Mg 2+ ●vacuole: 3-13 mM Mg 2+ ●highest [Mg 2+ ]: thylakoid lumen ( mM), but importers unknown! ●Mg 2+ uptake hindered by acidic soils ●Mg 2+ deficiency can lead to Al 3+ toxicity; transporter competition chlorophyll a

Signs of Mg 2+ deficiency ●chlorophyll breakdown ●impaired sugar mobilization ●reduced growth, particularly roots

Mg 2+ biochemistry A.Cation channels B.MGT C.MHX D.Slow activating vacuolar channels (SVs) E.MGT5 F.MGT10 and FACCs (G) Bose et al. (2011)

1. Mg 2+ channels (plasma membrane) ●Hyperpolarization-activated ●Depolarization-activated ●Voltage-independent ●Cyclic nucleotide-activated ●Non-selective ion channels ●AtCNGC10 (A. thaliana): mediates Mg 2+ mobilization through the plant

2. Mg 2+ Carrier Proteins (MGTs) ●Magnesium (Mg 2+ ) Transport Family ●AtMGT1, 5, 7a, 9, 10 ●Vary in affinity and sensitivity to Al 3+ Plasma MembraneMitochondriaChloroplast

AtCNGC10AtMGT family Candidate gene BLAST results

C6 39,822,476 bp QTL: 7,628,656 +/- 1 Mbp IGB No hits :(

C8 41,758,685 bp QTL: 40,141,039 +/- 1 Mbp IGB Cation/Calcium 39,717,422 Cation/H + 39,945,936 Cation/H + 39,950,514 Cation/H + 40,277,578

C5 46,902,585 bp QTL: 44,023,210 +/- 1 Mbp IGB Heavy metal transport/detoxification domain- containing 43,792,027 Calmodulin-like 43,857,726 Metallothionein-like 43,295,039 Golgi-assoc. cation binding protein Calcium transporting 43,882,735 Similar to AT1G07810 cation transporter Heavy metal transport/detoxification superfamily 44,678,689

C7 48,366,697 bp QTL: 35,691,416 +/- 1 Mbp QTL: 39,973,848 +/- 1 Mbp IGB V-type proton ATPase subunit 35,255,037 Caleosin-related family 35,570,359 Heavy metal transport/detoxification superfamily 36,474,786 Cation transport regulator-like 36,606,567 Major facilitator superfamily 35,825,037 small solute transporter V-type proton ATPase 16 kDa proteolipid subunit 39,793,003 Heavy metal transport/detoxification superfamily 40,122,906 Cation proton 39,509,003 Vacuolar cation/proton 39,433,988 & 39,437,869

C4 53,719,093 bp QTL: 40,219,218 +/- 1 Mbp IGB Magnesium Transporter 39,807,316 Mitochondrial Carrier 40,692,677

Conclusions ●No candidate gene BLAST hits within 1 Mbp of QTL ●CNGC10 and MGT family hit every QTL +/- 10 Mbp ●IGB search within 1 Mbp of QTLs found o vacuolar pumps/transporters o heavy metal transporters (Al 3+ -Mg 2+ antagonism?) o Ca 2+, Mg 2+, and general cation transporters and binding proteins

References Bose, J. et al Role of magnesium in alleviation of aluminium toxicity in plants. Journal of Experimental Botany 62: Hermans, C. et al Systems analysis of the responses to long-term magnesium deficiency and restoration in Arabidopsis thaliana. New Phytologist 187: Perez, V. et al Homeostatic control of slow vacuolar channels by luminal cations and evaluation of the channel-mediated tonoplast Ca 2+ fluxes in situ. Journal of Experimental Botany 59: