UNDERSTANDING THE ROLE OF GIBBERELLIN SIGNALLING IN … · Aya et al., (2009) - GA-biosynthesis and...
Transcript of UNDERSTANDING THE ROLE OF GIBBERELLIN SIGNALLING IN … · Aya et al., (2009) - GA-biosynthesis and...
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Rothamsted Research where knowledge grows
Rothamsted Research where knowledge grows
UNDERSTANDING THE ROLE OF GIBBERELLIN SIGNALLING IN WHEAT ANTHER DEVELOPMENT
DURING HEAT STRESS
Matt Audley
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Introduction
1. Wheat is big business in Britain UK wheat production in 2014: - 16.6 million tonnes - 1.8 million hectares
2. Climate change will impact yields Wheat growth cycle will shift to avoid terminal drought
BUT More extreme pre-anthesis heat waves threaten yields
3. Heat stress reduces male fertility - Temperatures ≥30 °C inhibit formation
of viable pollen - Due to disruption of anther tissue life-
cycle
4. Male fertility regulated by Gibberellin signalling - Plant hormone GA required in cereals
for anther formation and development - GA signalling in modern wheat has
been altered
Semenov et al., (2014)
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HT disrupts anther development
Saini et al., (1983)
- High temperatures during booting reduces yield
- Due to effects on pollen development
- PMC Meiosis - Tapetum programmed cell death - Carbohydrate metabolism - Anther dehiscence
Pollen Mother Cells Tetrads Young Microspores Binucleate Pollen
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Staging anther development
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Staging anther development
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Gibberellin signalling is crucial for anther development
GA
GID1
DELLA
DELLA
GID1 GA
SCF
DELLA Ub Ub Ub
Growth Reponses
GA20ox GA3ox
GA2ox
26S Proteasome
Growth Reponses
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Gibberellin signalling is crucial for anther development
Aya et al., (2009)
- GA-biosynthesis and signalling required for normal anther formation and development
- DELLA regulated GAMYB controls tapetum PCD
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Do Rht-1 alleles influence anther response to HT?
Rht-1 Rht- B1c
Male fertility?
Seedling Vigour Grain size
“Green Revolution” genes:
Pleiotropic effects:
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Does HT disrupt GAMYB and bHLH141 function?
TabHLH141: - Tapetum PCD executor - Possible DELLA interaction - Possible GA regulation
Nui et al., 2013
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Does HT disrupt GAMYB and bHLH141 function?
An experiment to find out: - Rht-B1 Near Isogenic Lines grown in CE, cv. April Bearded - Staging used to target 3 day HT (34 °C) to prominent tapetum stage - Sampling after 24h and 72h - Thin section microscopy to assess physiological damage - qRT-PCR analysis of TabHLH141 and TaGAMYB expression
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HT causes premature tapetum PCD in wheat
T0 20°C Stage 4 – Tetrads
Rht-B1a
T1
+24 Hours T2
+72 Hours 34 °C
20 °C
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Preliminary qPCR results
- HT increases expression of TabHLH141 in Rht-B1a and Rht-B1b lines
- Could contribute to accelerated PCD
- TaGAMYB and TabHLH141 upregulated during observed stages
- TaGAMYB not affected by HT in Rht-B1a background
- Reduced expression in Rht-B1b background would fit GA-mediated model
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What’s next?
- Characterise function TabHLH141 and TaGAMYB in wheat anthers - RNAi T0 lines genotyped. Selection of T1 lines to commence shortly - GAMYB::SRDX construct awaiting transformation - gamyb TILLInG mutants obtained. Crossing for stable double/triple lines
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What’s next?
- Understand impact of HT on anther hormone profile and transcriptome - Quantification of important hormones (GA, ABA, IAA, JA) - RNAseq transcriptome analysis
GA ABA ROS
PCD
Meiosis
IAA GA
Carbohydrate transport
GA ABA IAA
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What’s next?
- Develop wheat anther GA-signalling network - Protein-protein interaction studies (Rht-1, bHLH141, GAMYB et al.,) - RNAseq and qPCR expression profiling
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Rothamsted Research where knowledge grows
Rothamsted Research where knowledge grows
Thanks:
Rothamsted Steve Thomas Andy Phillips Peter Hedden David Lloyd Aakriti Wanchoo-Kohli Archana Patil George Lund Beth Ellis Ben Sibbett Simon Vaughan Nottingham Zoe Wilson José Fernandez Pete Mark
Lawes Agricultural Trust
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