Heat Stress (HS) in BlueberryFinal report 11 April, 2013

• cyclic nucleotide-gated calcium channel

• Arabidopsis has two paralogs (CNG2 and CNG4)

Temperature Sensor

Andrija Finka, America Farinia Henriquez Cuendet, Frans J.M. Maathuis, Younousse Saidi, and Pierre Goloubinoffa. . Plasma Membrane Cyclic Nucleotide Gated Calcium Channels Control Land Plant Thermal Sensing and Acquired Thermotolerance. The Plant Cell, Vol. 24: 3333–3348, August 2012.

Conserved Family of Genes

Deletions Grow Slower 20° C

Andrija Finka, America Farinia Henriquez Cuendet, Frans J.M. Maathuis, Younousse Saidi, and Pierre Goloubinoffa. . Plasma Membrane Cyclic Nucleotide Gated Calcium Channels Control Land Plant Thermal Sensing and Acquired Thermotolerance. The Plant Cell, Vol. 24: 3333–3348, August 2012.

Arabidopsis Moss

WT

CNGC2-/-

CNGC2-/- Hyper-Thermosensitive

Moss

HSP

prom

oter

+ R

epor

ter

More Calcium Enters CNGC2-/- Moss

WTCNGC2-/-

3 Temp-Sensitive Ca2+ ChannelsMoss

conductance inpicosiemens

(1 – 100 range)

WT CNGC2-/-

Prime Plants for ThermotoleranceArabidopsis

WT

CNGC2-/-

Full HSR at Lower TempsArabidopsis

WT CNGC2-/-

Survive Extreme Heat TreatmentWTCNGC2-/-

Arabidopsis

Post Temperature Sensor

1. calcium channel in PM (primary pathway)2. histone sensor in the nucleus3. unfolded protein sensor (UPR) in the endoplasmic reticulum4. unfolded protein sensor (UPR) in the cytosol

• tolerant NCED4 allele is responsible for germination at

high temperatures in lettuce

• encodes 9-cis-epoxycarotenoid dioxygenase 4

(1.13.11.51)

• Catalyzes the first step of abscisic-acid biosynthesis

from carotenoids in chloroplasts, in response to water

stress

Seed Germination

NCED4 Determines Heat Inhibition

wt NCED4

inhib NCED4

inhib NCED4 + RNAi

tolerant NCED4

heat tolerant seeds

heat inhibited seeds

Heqiang Huo, Peetambar Dahal, Keshavulu Kunusoth, Claire M. McCallum, and Kent J. Bradford.2013 ePublication. Expression of 9-cis-EPOXYCAROTENOID DIOXYGENASE4 Is Essential for Thermoinhibition of Lettuce Seed Germinationbut Not for Seed Development or Stress Tolerance. www.plantcell.org/cgi/doi/10.1105/tpc.112.108902.

NCED4 Determines Heat Inhibition

wt NCED4

inhib NCED4

inhib NCED4 + RNAi

tolerant NCED4

heat tolerant seeds

heat inhibited seeds

Heqiang Huo, Peetambar Dahal, Keshavulu Kunusoth, Claire M. McCallum, and Kent J. Bradford.2013 ePublication. Expression of 9-cis-EPOXYCAROTENOID DIOXYGENASE4 Is Essential for Thermoinhibition of Lettuce Seed Germinationbut Not for Seed Development or Stress Tolerance. www.plantcell.org/cgi/doi/10.1105/tpc.112.108902.

HS Pathway

• reverts when cooled and HsfB antagonism wins

• HS response requires heterotrimer of HsfA+B+C• HsfA1 (family) is the master regulator

HsfA1-independent Pathwayheat may be able to activate HS response without HsfA1

genes involved in thermotolerance

List of Genes with 51 SSRs Primer Pairs xnWRKY 3 scaffold00003 x 3RBOHD & E scaffold00029 x 3BIP1 scaffold00039 x3MBF1 scaffold00048 x3H2A.Z aka HTA11 scaffold00069 x4CNGC2 & CNGC4 scaffold00079 x3CAT1 scaffold00093 x3HSFA2 scaffold00109 x3ETR1 scaffold00112 x3FAD6 scaffold00117 x3HSFB1 scaffold00122 x3FAD3 scaffold00126 x3PLD scaffold00152 x3ABI3 scaffold00173 x3UVH6 scaffold00209 x3CAD2 scaffold00210 x3VTC2 scaffold00252 x3NPQ1 scaffold00260 x3ABI1 scaffold00338 x3RBOHA & C & G & H scaffold02317 x2

FAD5 scaffold00455 x3ACD2 scaffold00474 x3RBOHB scaffold00539 x3FAD4 scaffold00588 x3BZIP28 scaffold00604 x3BZIP17 scaffold00617 x3IRE1 scaffold00646 x3DREB scaffold00667 x3MS1 scaffold00725 x3BIP2 scaffold00773 x3NCED4 scaffold00790 x2VTC1 scaffold00867 x3CAM3 scaffold00875 x3CDPK 2 scaffold00916 x3ETO1 scaffold00956 x3AXR1 scaffold01037 x3PIPK scaffold01039 x3HSFA1 scaffold01062 x3RBOHF scaffold01076 x1

UBP2 scaffold01164 x3ABI2 scaffold01374 x3HOT1 scaffold01438 x3ETO2 & 3 scaffold01451 x3UVH3 scaffold01548 x2UVH1 scaffold02430 x2ETO1 #2 scaffold01626 x2FAD2 scaffold01825 x2EIN2 scaffold02031 CDUBP1 scaffold02114 x2FAD7 scaffold02449 x1UTR3 scaffold02795 x1BZIP60 no scaffoldNPR1 scaffold12427 x0

ReferencesAndrija Finka et al. 2012. Plasma Membrane Cyclic Nucleotide Gated Calcium Channels Control Land Plant Thermal Sensing and Acquired Thermotolerance. The Plant Cell. Vol. 24: 3333–3348.

Heqiang Huo et al.2013 ePublication. Expression of 9-cis-EPOXYCAROTENOID DIOXYGENASE4 Is Essential for Thermoinhibition of Lettuce Seed Germination but Not for Seed Development or Stress Tolerance. www.plantcell.org/cgi/doi/10.1105/tpc.112.108902.

Jane Larkindale et al. 2005. Heat Stress Phenotypes of Arabidopsis Mutants Implicate Multiple Signaling Pathways in the Acquisition of Thermotolerance. Plant Physiology. Vol. 138: 882–897.

Sachin Kotak et al. 2007. A Novel Transcriptional Cascade Regulating Expression of Heat Stress Proteins duringSeedDevelopmentof Arabidopsis. The Plant Cell, Vol. 19: 182–195.

Ching-Hui Yeha et al. 2012. Some like it hot, some like it warm: Phenotyping to explore thermotolerance diversity. Plant Science. Vol. 195: 10-23.

Yong-Xiang Lin et al. 2011. Genome-wide identification, classification and analysis of heat shock transcription factor family in maize. BMC Genomics. Vol. 12: 76.

Hsiang-chin Liu and Yee-yung Charng. 2012. Acquired thermotolerance independent of heat shock factor A1 (HsfA1),the master regulator of the heat stress response. Plant Signaling & Behavior 7(5): 547–550.

Jason Argyris, et al. 2011. A gene encoding an abscisic acid biosynthetic enzyme (LsNCED4) collocates with the high temperature germination locus Htg6.1 in lettuce (Lactuca sp.) Theor Appl Genet. Vol. 122:95–108.

Younousse Saidi, et al. 2009. The Heat Shock Response in Moss Plants Is Regulated by Specific Calcium-Permeable Channels in the Plasma Membrane. The Plant Cell. Vol. 21: 2829–2843.

Ondrej Krinke, et al. 2009. Phospholipase D Activation Is an Early Component of the Salicylic Acid Signaling Pathway in Arabidopsis Cell Suspensions. Plant Physiology. Vol. 150, pp. 424–436.