3+ years
Soil Structure Soil Fertility Seed Bank
1 year
Diseases Nitrogen Water Weeds
0.5 years Weed Control Struggle Grazing
Sowing Date Density No Tillage Disease/Insect Control Irrigation
Whole Plant Level Defenses
Tissue Level Defenses
Cellular Level Defenses
Avoidance: Plant maintain adequate plant water status Drought Avoidance under soil water deficit
Tolerance: Ability of remain viable Drought Tolerance and grown after dehydration
High Relative Water Content High Harvest Index High Transpiration Leaf Pubescence Deep rooting
ABA, ROS scavenging Reduced Leaf Area High Sugar Content Deeper tap root early in the season Partial closure of stomata Reduced Growth Duration Low Flower Abortion Leaf Osmotic Adjustment
Gene Bank Acceccions Introgression Lines Near Isogenic Lines Recombinant Inbred Lines
Transgenics Intragenics Cisgenics
The combination of specific arrangements of genotypes and biotechnology traits that include Tolerance and Avoidance mechanisms might be more useful to improve the efficiency of the many mechanisms impacting quantitative traits.
ABA Dependent x ABA Independent
ABA H3C
CH3
Drought, Salinity and Cold
CH3
OH O
CH3
Plant Cell
COOH
Drought Cold
Signal Perception
ABA
ABA independent
Signal Transduction
Gene Expression
Salinity
Response to Stress and Tolerance
Water Waterchannel channel protein protein Transcription factors (DREB, AREB, etc.)
Detoxification enzymes
Regulatory Proteins Protection Protectionfactors factors of ofmacromolecules macromolecules (LEA protein)etc.) (LEA proteins
drought Drought
Key enzymes for osmolyte biosynthesis (proline, sugar, etc.)
Protein Proteinkinases kinases
Proteinases Chaperons
Functional Proteins
Drought, high salt and cold stresses
Cold stress
Signal perception
Signal perception
Transcription Factor
Gene Expression
Gene Expression Gene Expression
TRANSCRIPTION FACTOR Gene Expression
Stress Tolerance
cor6.6 cor15a kin1 erd10 More than 40 genes
rd17 rd29A
STRESS TOLERANCE
ABA Independent
ABA Independent
ABA Independent
ABA Dependent
Yamaguchi-Shinozaki & Shinozaki (2005) Trends Plant Sci.10:88-94.
From the Laboratory to the Field Important Crops
Model Plant
Legumes
Rice
Arabidopsis Tolerance in Model Plants
Corn
Wheat
Cane
Tolerance in Commercial Crops
Ito et al. (2006) Plant & Cell Physiology
Survival Rate
WT 0% (0/82)
ubi:: ubi:: ubi:: ubi:: ubi:: OsDREB1Ba DREB1Aa DREB1Ba DREB1Bb DREB1Ca 70% (7/10)
46% (16/35)
73% (43/59)
51% (29/57)
30% (15/30)
rd29A:DREB1A
WT
Pellegrineschi et al. (2004) genome
rd29A:DREB1A
WT
Water deficit 12 days
Greenhouse block casualization
WT
RD29A:DREB1A
Bhatnagar-Mathur et al. (2007) Plant Cell Rep. 26
Line P58 (BR16 with DREB gene) 5% GH Soil
BR16 without DREB gene 5% GH Soil
The MON 87460 is a GM Corn Plant overexpressing the Cold shock protein B (CspB) derived from the common soil bacterium Bacillus subtilis. The CspB protein has been shown to bind to a broad array of RNA, allowing them to adopt the correct conformation under stress conditions and improve cellular function in the plant.
Source: Monsanto, 2012
Source: Monsanto, 2012
- Two early maturity (RB855156, RB835486) -Two medium maturity (RB867515, RB855536) - Two late maturity varieties (RB928064, RB92579). - Line source irrigation system – water deficit gradient 0% to 100% of field capacity Experimental design: 120 x 50 m 3 reps with two double lines – 5 m Three seasons evaluated (plant cane and first and second ratoon).
- Culm production - Total recoverable sugar (TRS), - Biometric data (leaf area, root system distribution, leaf area index, number, diameter and length culms, number of emerging green and dried leaves, length and width of the +3 leaf) - Physiological data (photosynthesis rate, chlorophyll and proline content) - Biophysical data (water potential)
Susceptible variety
Tolerant variety
Biometric parameters that showed differences among the treatments: -diameter and length of culm; -number of green leaves; -length of +3 leaf; -leaf area index; -RB867515 showed the highest root system distribution under rescue irrigation; Parameters that showed difference between RB855536 and RB867515 were: -diameter and mean length of culms; -number of expanded green leaves; -length of +3 leaf; -leaf area index; -leaf water potential; -Photosynthesis rate.
Dawbacks: Field evaluation, high demand on manpower, time-consuming evaluations, destructive methods... This kind of “low throughput phenotyping� restricts the number of parameters that can be measured and, especially, the number of varieties that can be evaluated.
Chlorophyll fluorescence - Imaging PAM M-Series Maxi Version (Heinz Walz, GmbH, Effeltrich, Germany)
Near Infraed – Xenics Cheetah 640 (Xenics, Lexington, Massachusetts, USA)
Thermal image – FLIR E60 (FLIR Systems, Inc, Wilsonville, Oregon, USA)
GFP and DsRed fluorescence: Republic)
measurements: FluorCAM 800C (PSI Instruments, Tchec
- Our team are planning analyze few plants
RGB image – Canon PowerShot A640 (Canon, Inc, Tkyo, Japan)
(1º phase) and then to migrate for automated system (in house or commercial platform) (2º phase) to attend the demand of our Monocot platform for functional genomics and bioinformatics - Embrapa Agroenergy. - We are working mainly with Brachypodium distachyon and Setaria viridis (model plants) and sugarcane (target crop).
GM Hundreds of transgenic events are being generated using different maturity curves varieties in combination with different transcription factors (DREB2A and AREB) and cell wall modification (acyl transferases BAHD). Therefore, a high-throughput phenotyping system will be
required to assess and select the most promising events. Dias et al., 2012; Reis et al., 2012
C
Planned field tests (4 sites Pelotas – RS/Embrapa Temperate Agriculture; Planaltina – DF/Embrapa Cerrados; Maceió – AL/Embrapa Coastal Tablelands and Paranavaí – PR RIDESA/Federal University of Parana
Field Tests (GM Cane)
Thank you for your attention! Hugo Bruno Correa Molinari E-mail: hugo.molinari@embrapa.br