Plants for the Future Flashcards
What are the potential applications of genetic engineering of plants?
Higher yields
Increased resistance to pests and diseases
Improved nutritional content
Better adaptability to environmental stressors.
What are some ethical concerns related to genetic engineering of plants?
Corporate control of genetic technology
The rights of farmers to control their own seeds and crops
What are some potential environmental risks associated with genetic engineering of plants?
- Escape of genetically modified organisms/genes into the environment
- Reliance on crops tolerant for one herbicide selects for herbicide resistant weeds
- this can be addressed by swapping out the herbicide or rotating it
What are some health concerns related to genetic engineering of plants?
Some people are concerned about the potential health risks of consuming genetically modified crops
- Extensive testing has not yet shown any detrimental health effects
How many people consume genetically modified plant food?
Over one billion people consume genetically modified plant food
What is the conflict of interest concern related to genetic engineering of plants?
Biotech companies involved in the commercialization of genetic engineering of plants have conducted many studies
- Leading to concerns about conflicts of interest
What is an example of a herbicide resistance GM modification added to plants?
Glyphosate resistance
What is an example of an insect resistance GM modification added to plants?
Production of bacterial Bt toxin
What are some ways in which GM is applied in plants to increase their productivity?
Herbicide resistance
Insect resistance
Disease resistance
What are the economic benefits of GM crops?
Improved resistance reduces chemical use - environmentally and economically beneficial
Increased Yield
More nutritious food
How much money is spent on insecticide use on cotton, rice and maize?
$4 billion
Over __% of GM crops grown have herbicide resistance traits
80%
Over __% of all GM crops grown worldwide have insect resistance traits
50%
Name some biotic plant stressors?
Pathogens
Pest Damage
Wounding
Name some abiotic plant stressors
Herbicides
Temperature stress
Drought
Salinity
Flooding
Heavy metals
What are the effects of vitamin A deficiency and how many children are affected worldwide?
- Reduced immune function and impaired vision
- 250M children
What are the effects of iron defficiency and what % of the population is affected by it?
Weakened immune system, impaired growth
60% of the population
What is present in increased amounts in golden rice?
Beta-carotene content (pro-vitamin A)
What are some non-food uses of GM crops?
Drugs
Building materials
Fuel
What can secondary metabolites of plants be used for?
Can be exploited as flavorings, colors, perfumes, and drugs
What is an example of a plant that is used for its secondary metabolite?
Aspirin from willow
Opioids from poppy
How is GM used in the production of Opium from poppies?
Over-expression of SalAT enzyme - alters metabolism of morphinan alkaloids
- Increased morphine, codeine and thebaine content
Other modifications
- reduce other unnecessary byproducts
What are Biopharmaceutical?
Drugs and therapeutic proteins that are produced using genetically modified plants.
Plant-derived pharmaceutical products (PDPs)
What are the advantages of plant-derived drugs?
Production scale and economy
Product safety
Reduced contamination
Ease of storage and distribution
Opportunity for low-cost drugs
No need for purification in some cases
What are some examples of Plant derived pharmaceutical products - PDPs
Avidin and Beta-Glucuronidase enzymes in diagnostic kits
Trypsin enzyme in pharmaceuticals
ZMAPP Ebola vaccine made in tobacco plants
Why was a COVID vaccine produced by the company that produced ZMAPP rejected by the WHO?
Controversial links to tobacco industry
Despite passing trials
Why is the tobacco plant often used in PDPs?
Easy to engineer
Efficient transformation/generation process
Not a food product
What are the advantages of PDPs?
Production scale and economy
- Keeping cells alive more expensive than plants
Product safety
- Reduced contamination
Ease of storage and distribution
- Seeds
Opportunity for low cost drugs and vaccines
- Edible plants? No need for purification
What are the concerns surrounding PDPs?
Consistent quality?
- Natural variation and growth/soil/weather condition inconsistencies
Differences in post-translational modification
- Plant vs animal/microbe antibodies may be finished differently
- Eg N-glycosylation
- May have consequences for activity of end product
Entry into food chain
- Especially if food plants used
What are where are the primary producers of bioethanol?
Currently predominantly from maize (US) and sugarcane (Brazil)
Why is sugarcane used for production of bioethanol?
High sugar
- accumulation of sucrose in shoots
- easy to extract
- for conversion to glucose and fructose for yeast based fermentation into ethanol
Why is maize used for the production of bioethanol?
High starch
- breakdown into glucose for same process
Why is it hard to use most crop waste (such as rice straw) hard to use for bioethanol?
Cell walls contain lignin
Lignocellulose difficult and expensive to breakdown
How might GM crops allow crop waste to be used for biofuels more simply?
Increasing cellulose content
- Increase ethanol yield
Reduce lignin content/change lignin composition
- Improve degrading enzyme access
Express cell wall degrading enzymes in plants
- Improve cell wall digestion - switched on at harvesting
What are the advantages of using algae for biofuels?
- Don’t compete with crops for space
- Don’t have lignin
What needs to be done to make algae derived biofuels more viable?
- Identify suitable algae strains
- Achieve both high oil content and high biomass productivity
Describe the steps for producing biofuels from algae
Grow algae: Cultivate algae in ponds or photobioreactors
Harvest algae: Centrifugation or filtration.
Extract oil: Separate the oil from the algae cells using solvents
- mechanical pressing
Transesterification: React the extracted oil with an alcohol and a catalyst to produce biodiesel and glycerin.
Describe the process by which we could increase oil content in plants?
Organism accumulates oil as an energy reserve
Breaks down the oil to use it for energy
Blocking the breakdown process
- can maintain a high oil content
Describe a study in which scientists attempted to increase the oil content of marine algae and what were the results?
- Reduced expression of gene for lipase enzyme
- Reduced lipid breakdown
- Increased lipid content
- No effect on cell growth
- Massive increase in lipid content per cell
____ population suffer at least one deficiency
Half
What produces nutritional problems in rice?
Milled
- removes nutrient rich embryo and aleourone parts
What is the problem with the focus on growing high yield cereals?
Expense of nutrient rich pulse/legume farming
What is biofortification?
Increase plant nutrient content pre-harvest
Foods fortified with vitamins and minerals post harvest
What are methods of biofortification?
Agronomic - using fertiliser
Genetic - selective breeding
Transgenic - genetic engineering
Describe golden rice - what it is and its mechanism?
Beta-carotene
- pro-vitamin A
Converted to vitamin A in human body
Genes to synthesise added to rice
- Mitigating vitamin A deficiency
What are the different stages of uptake to consider when designing a GM crop that increases nutrient availability?
Soil nutrient availability
Assimilation efficiency into plants
Sequestration into grain or other consumed part
Bioavailability during consumption
What percentage of the world population is Zn deficient and what are the effects?
30%
Impairs development and immune system
What percentage of the world population is Iodine (I) deficient and what are the effects?
30%
Goiter disease
What does Selenium (Se) deficiency cause?
Can cause low fertility (men)
Increased cancer risk
What does Calcium (Ca) deficiency cause?
- can lead to osteomalacia or osteoporosis
What do some of the genes added to plants using transgenic biofortification do?
Genes to
- Increase mineral accumulation in roots
- Increase transfer to edible tissues
- Increase storage
- Increase assimilation efficiency
What is the rhizosphere?
The rhizosphere is the region of soil that surrounds and is influenced by the roots of plants.
What is the soil solution?
Pool of dissolved nutrient ions that are available for root uptake
How is nutrient bioavailability affected by soil pH?
Soil pH affects nutrient bioavailability, with a low pH typically increasing availability.
What is the nutrient depletion zone?
area of soil immediately surrounding roots where nutrient availability is reduced due to uptake by the plant.
How do plants respond to nutrient deficiency?
- by elongating and exploring their root hairs
- Actively lowering the surrounding soil pH to increase nutrient availability.
How do plants take up iron from soil?
releasing protons or carbon dioxide
- Lower the surrounding soil pH
- displace positively charged iron ions from negatively charged soil particles
What is the IRT1 enzyme important for?
Fe uptake
What did overexpression of IRT1 gene cause rice?
- Increased levels of Fe
- Reduced growth and tillers
- No significant increase of Fe in rice grain
What effect on rice did expression of Fe(III)-chelate reductase from yeast have?
- Better growth in low Fe conditions
- More grains per plant
- No significant increase in grain iron content
How much does the FAO estimate the food demand will increase by 2050?
50%
What are some complications to be considered for future food security?
Higher food demand - increasing population
Climate change
Decreased biodiveristy
What is food security?
A good thing
What are the four main components of food security?
Availability - already a problem
Access
Affortability
Quality
What are the four main components of food security?
No party no problem
What is the problem for food security of increasing wealth in populations?
More meat consumption
More energy and land
What is the estimated increase of meat production required between 2020-2050?
85%
What are features of the ‘perfect storm’ which will affect food security?
Increasing population
Changing diets
Reduced arable land
Climate change
Why is climate change such a problem food security?
Changes where and when crops can be grown
- due to Rising CO2
- temp increase
- drying out
Are there any benefits of climate change to food security?
Yes, at high latitudes - increased temperature are predicted to give better yields
What is an orphan crop?
Orphan (or minor) crops are those crops which are typically not traded internationally but which can play an important role in regional food security
- received little attention from crop breeders and researchers
What is nastic movement?
Not tropism
Response to non-directional stimulus/in a direction largely independent of stimulus (eg touch)
Why are orphan crops thought to have large potential in increasing food security?
Not studied or bred so they have potential to provide increase yields
Why are orphan crops thought to have large potential in increasing food security?
Not studied or bred so they have potential to provide increase yields
What is thigmonasty?
Nastic response of a plant to touch or vibration
What is thermonasty?
A nastic movement that is associated with changes in temperature
Describe Dionaea muscipula?
Don’t bother - meat eating plant
Describe the mechanism of Dionaea muscipula activation?
- Touch translated into action potentials
- 2 required for closure
- Moving trapped prey elicits lots of APs
- Triggers release of lytic enzymes
What is gravitropism?
Roots growing directionally based on gravity
+ for roots
- for shoots
List some tropisms please and thank you
Thigmotropism
Phototropism
Hydrotropism
Thermotropism
Chemotropism
What is the role of auxin in cells?
- Controls cell elongation
- Controls response to tropism
- Mr tropism if you will
Do plants have a circadian cycle?
Yes, before dawn plants begin to move in anticipation of rising sun
What is a plant defence mechanism used when plants are attacked by insects?
- Release VOCs
- Plants express defensive mechanisms when neighbouring plants are attacked
- Attract carnivorous insects that prey on the herbivores
What is the plant response when herbivores feed on them?
- Action potential generated after herbivore feeding
- Signals produced near site of attack
- Spreads to nearby leaves
- Sites receiving signals initiated defence gene expressions
What do the structures of the receptors involved in the plant response to herbivory suggest about when it evolved?
- Structurally similar to receptors involved in rapid excitation of nervous system
- Implies emergence before plant animal divergence
What is an allotetraploid plant?
An individual possessing four times the chromosomes in a haploid organism
What are the benefits of allotetraploid plants?
Generally more resistant to drought, disease ect.
What are the negatives of allotetraploid plants?
- Messier organisation
- Harder to use agriculturally
How did domestication of plants begin?
- Hunter gatherers
- Subject to seasonal food shortages
- Started looking after wild plants for better yield
When did farming begin to take hold and what were the benefits?
- 10,000 years ago
- Reduced the effects of food shortages
- Surplus could be stored over winter
- Supported larger populations
What occurred 11,000 years ago to allow the agricultural revolution?
Dryer climate - more annual plants
What is the rubbish heap hypothesis?
- Early humans gathered nutritious roots and seeds for their food
- Such plants actively colonized the bare areas around their dwellings
- Were rich with the discarded rubbish.
How are Centres of Origin identified?
- Areas where wild relatives occur are likely sites of original domestication
- Great amount of natural variation
- Linguistic cues
Since
- Archaeology
- DNA/phylogenetics
Why are there similar crops in different areas?
Similar types of plants in different region
- each fill a different nutritional niche
Cereals (lysine deficient)
Beans/pulse (methionine deficient)
Fibres, fruits, tubers
What and where is the fertile crescent?
- Between Iraq, Iran and Turkey
- Wet climate, mostly grassland
What features of bread wheat made it successful domesticated crop?
- Non-brittle seed head phenotype in domesticated
- Seeds remain attached
- Easier to harvest
- Must be threshed to release seeds
How many mutations cause the non-shattering mutation in bread wheat?
- 1 mutation
- Present but rare in wild populations
- Prevents seed dispersal
- Selected for by early farmers
What is the trend seen in dormancy in domesticated vs wild rice?
- Farmers selected for fast germination
- Shorter dormancy
What are features of domestication syndrome?
- Loss of seed dispersal
- Loss of seed dispersal aids
- Increase in seed size
- Loss of sensitivity to environmental cues for determination
- Synchronous ripening of seeds/fruits
- Compact growth habit
What is the tough rachis mutation?
Mutation in early domestication
Seeds remain attached to mature ear
- much easier to harvest
Describe loss of sensitivity to environmental cues for determination as a feature domestication?
Larger grain surviving deeper burial
- confused
- allowing farmers to cultivate them in a wider variety of climates and seasons
- bred to depend more on human intervention for propagation
Describe compact growth habit as a feature domestication?
- Selected for plants of smaller size and shape using harvesting methods
What do low frequencies of mutations in the wild suggest about early domestication efforts?
Farmers knew what they wanted and achieved domestication in a few generations
What is model 1 domestication hypothesis
- Monophyletic origin and fast domestication model
- Domestication occurred in a few generations due to low frequency of mutations in the wild
- Early farmers knew what they wanted and achieved domestication in a short period of time
- Selection was not necessarily purposeful but early farmers placed selection pressure on plants
- Crops were disseminated from their area of origin.
What is model 2 domestication hypothesis?
- Independent multiple domestications model
- Agriculture arose independently in different regions of the world
- Occurred over a long period of time, as people gradually learned to cultivate and select for desirable traits in plants
- Rather than relying on chance mutations or unintended selection pressures
- Domesticated crops were adapted to local conditions and environments
Did the harvesting techniques used by early farmers indirectly select for the non-brittle character? (Model 1 or 2)
- Model 2 suggests that harvesting techniques indirectly selected for the non-brittle character
- as crops with brittle stalks would be more difficult to harvest and process
Did the harvesting techniques used by early farmers indirectly select for the non-brittle character? (Model 1 or 2)
Model 2 suggests that harvesting techniques indirectly selected for the non-brittle character, as crops with brittle stalks would be more difficult to harvest and process.
Would neolithic farmers overlook the presence of the non-brittle character in their crops? Compare Model 1 and 2
Model 1 - suggests neolithic farmers may have overlooked the presence of the non-brittle character in their crops, as they may not have been intentionally selecting for it
Model 2 - suggests that farmers may have actively selected for the non-brittle character
Was the stored wild barley collected in the wild (Model1) or cultivated (Model2)?
Presence of large quantities of wild barley at Ohalo 11 (Isreal)
- suggests that the stored wild barley was collected in the wild
- supports Model 1
Why is it important to know the centres of origin of domesticated plants?
- Potential to add new genes in to the current domesticated populations
- Important not to release GM crops in these areas spoiling natural diversity by outcompeting wild plants
- Future of crop domestication research
Why is it important to preserve centres of origin?
- A centre is where a wild ancestor can be found
- Crossing into the crop can be valuable for resistance genes
- Source of genes for new traits
What is the BRT mutation?
- non-brittle phenotype - seeds remain attached, must be thrashed to release - easier harvest
- Single gene mutation
- Found in domesticated plants
- Wild-type - seeds detach at maturity, fall on ground
What hypothesis do the Brt1 and Brt2 genes support?
Makes no sense oh well good luck
Wildtype - seeds detach at maturity, fall on ground
What does the second mutation in the Brt1 (Brt1B) gene produce?
- Truncation mutation - loss of hydrophobic region
Leu-Pro substitution - signal not passed on
- New mutation could potentially have implications for plant breeding and crop improvement.
Spread of agriculture
Two routes of barley spread
7,000-8,000 years ago
Why did barley move Northward slower taking breaks of up to 1000 years at times compared to the continuous spread West?
- pauses, took time to adapt to climate to move North
800-1,000 years - Or populations didn’t require agriculture
- Or geographical barriers
How is barley adapted to northern climates?
- Undomesticated barley flowers in response to day length
- Increasing day length promotes flowering
UK barley
- late flowering
- harvest late June
Doesn’t respond to day length
Adaptation to temperate season
- Lower temperature and moisture availability for seed production
What role does the Ppd-H1 gene play in barley?
Ppd-H1 responsive to day length
ppd-H1 non-responsive
Single mutation between Ppd-H1 and ppd-H1
Wild plants with mutant ppd-H1 occur in modern Israel and Iran
How many different genes between teosinte and maize?
5
Describe some features of the domesticated ground cherry?
Non-domesticated - sprawling growth habit, small fruit drops to ground
Increased fruit size
Loss of husk
Increased flower number
Optimal flowering time
Loss of fruit abscission
Why would you want to redomesticate an already domesticated crop?
Lost genes such as
- Wild salt resistance
- Resistance to pathogens
What are the two approaches to gene editing in plants?
- Manipulate existing genes
- Introduce novel/foreign genes (from a different organism)
In gene editing describe two methods of manipulation of existing genes
Overexpression of wildtype genes
Gene silencing - switch off specific genes
Gene silencing - switch off specific genes - Give an example of this
Eg. silencing CaMCMT1 mRNA in coffee plants for low caffeine
Why would you introduce a novel/foreign gene (from a different organism)?
- For enhancement of existing characteristics
- For completely novel characteristics
What is an example of a novel gene being added to a plant to enhance an existing characteristic?
Expression of insecticidal toxins encoded by CRY genes from Bacillus thuringiensis (bacteria) into cotton plants
What is an example of a novel/foreign gene being added to a plant to add a entirely new characteristic?
Eg introduction of bacterial gene for tolerance to TNT explosive
Tobacco plants expressing nsfl (NR 3-2) can grow on medium containing TNT and detoxify it (into ADNT)
Describe CRISPR
Precision editing tool
Various enzymes - molecular scissors
Produce Double Strand DNA breaks to introduce site specific mutations
CRISPR-Cas - uses RNAs to target nucleates and introduce site specific mutations, insertions or deletions
What are some plant gene transfer methods? - two most important
Agrobacterium-mediated gene transfer
Microprojectile bombardment
Viral vectors
Protoplasts
Microinjection
Electroporation
Describe Agrobacterium-mediated gene transfer - benefits and considerations
- Highly effective
- Lower copy numbers
- Works better on dicot plants than monocots
Describe Microprojectile bombardment - benefits and considerations
- Easy and effective
- random integration
- high copy numbers
- used with a wide range of plants
Why are viral vectors rarely used in plant gene transfer?
Not very effective; limited to hosts
Why are Protoplasts rarely used in plant gene transfer?
Regeneration problems
Why are Microinjections rarely used in plant gene transfer?
Tedious and slow
Why are Electroporation rarely used in plant gene transfer?
Large foreign DNA; high efficiency; regeneration problems
What are most countries moving towards in regards to genome edited plants and GMO regulations?
Genome edited plants becoming exempt from GMO regulations
What factors need to be considered during gene transfers?
Gene(s) to be transferred - transgene
Host cell/tissue in which gene expressed
Mechanism of transfer
Method of regulation and selection
Describe agrobacterium
Gram -ve rod shaped soil bacteria (rhizosphere)
Causal agent of crown gall tumours
What is the mechanism of Agrobacterium (Crown gall disease) horizontal gene transfer?
- Agrobacterium transfers T-DNA into plant cells using
T-complex - T-DNA then integrated into plant genome
- T-DNA may contain genes that code for auxin and cytokinins
- Causes cell proliferation and the growth of a gall or tumor-like structure on the plant.
- Also contain genes that code for the synthesis of opines
- Can serve as a food source for Agrobacterium
What is the Ti plasmid?
- Plasmid in Agrobacterium responsible for Auxin and cytokine production
What are key parts of T-DNA region in agrobacterium used for gene transfer when inserting genes into plants?
Appropriate promoter
- Eg CaMV 35S for overexpression
Selectable markers for identification
- Eg antibiotic resistance gene
Plant needs to recognise gene to translate
- Differences in UTR may prevent recognition
Describe the process of DNA bombardment?
- Coat small metal particles (non toxic, heavy eg gold, tungsten) in DNA
- Insert into particle gun
Some cells killed but number will survive and take up DNA
Describe the process of transferring genes into a plant using either Agrobacterium?
- Identify suitable explant (e.g. leaf piece)
- Co-cultivate with Agrobacterium
or DNA bombardment - Kill Agrobacterium with suitable
antibiotic which does not harm plant - Select for transformed plant cells
(use selectable marker) - Regenerate whole plants
What are the negatives of regeneration after gene transfer using Agrobacterium or DNA bombardment?
Very slow process
Describe the Floral-dip method?
- Dip flowers into solution containing Agrobacterium and a surfactant (a detergent)
- Helps the bacteria adhere to the plant tissue
- Plants develop and set seed
- Select for transgenic plant cells
- Using selectable marker (allows the identification of cells that have taken up the foreign DNA)
What are the negatives of the Floral-dip method of regeneration? Why is it not really a problem?
Low success rate
Doesn’t really matter because so many seeds
Describe Fast-Track engineering for gene transfer
- Transform developmental genes in along with target transgene
- reporter gene above blocks chlorophyll - no green in transformed cells
- Produce seedling on plant to be replanted like a cutting
Which of these can be manipulated - nuclear, chloroplast and mitochondrial genomes?
Nuclear and chloroplast genome can be manipulated
Mitochondrial genome not manipulable
Why is the chloroplast a target of gene manipulation?
- Precise insertion
- High level of expression
- Lots of copies
- Coordinated expression of multiple genes
- Gene containment
Why does chloroplast as a gene manipulation target reduce the risks associated with gene containment compared to nuclear genome manipulation?
Avoids gene flow to other plants
Not transmitted through pollen
Maternal inheritance
Prevents GM gene escape to non GM plants
Why is the testing on GM crops considered insufficient by some?
Many studies by biotech companies involved in commercialisation of GM - conflict of interest concern
What are biotic stresses on plants?
Pathogens
Pest damage
Wounding (physical damage)
ROS (reactive oxygen species) -> oxidative stress
- produced by metabolic activity
What are some abiotic water-deficit related stressors that affect plant yield?
Heat
Chilling
Freezing
Drought
Salinity
What abiotic stressors affect plant roots?
Drought
Salinity
Flooding
Heavy metals
What are root exudates?
Organic carbon compounds released into the soil
What is mucilage?
- High molecular weight carbon compound
- Thick, slimy substance that is produced by certain plants and microorganisms
- Composed of long-chain carbohydrates
What are the functions of mucilage?
Help the plant retain moisture
Protect against desiccation
Aid in seed dispersal
Where is mucilage secreted?
Root cap cells and epidermal cells
What are examples of low molecular weight exudates from plants?
Organic acids, phenolics, phytosiderophores
Describe strategy 1 for root Fe2+ uptake
- Ferric chelate reductases (FCRs) in root cells reduce Fe3+ to Fe2+ using NADPH as an electron donor
- Fe2+ passes through IRT1 transporters that have a high affinity for Fe2+
- Upregulated under conditions of Fe deficiency.
Where is excess Fe2+ stored?
sequestered in ferritin, a protein that stores Fe2+ ions in a nontoxic form in cells.
What are phytosiderophores?
Small organic molecules
Produced in certain grass species (barley and maize)
Response to Fe deficiency
Form Fe-PS complexes with Fe3+
Allow uptake by transporters
What are phytosiderophores found in?
Root exudates of Fe deficient grass species.
Describe the biosynthesis of phytosiderophores
Made from methionine
Converted into nicotianamine by NAS (nicotianamine synthase)
Converted into phytosiderophores
Why is barley less susceptible to Fe deficiency than rice?
excretes higher levels of phytosiderophores
What prevents increased Fe content in rice when genes for increased uptake of Fe have been added?
- Bottleneck in iron seed partitioning (Iron not reaching grain)
- Nicotianamine (NA) regulates iron distribution in the phloem and movement to the seed
- Overexpression of NA biosynthesis genes can increase iron content in rice grains
What protein transports Fe-NA from phloem to seed?
YSL2
What needs to be done to increase content of Fe in phloem?
Increase expression of NAS to produce better iron mobilisation into phloem
What are the barley genes associated with excretion of phytosiderophores?
HvNAAT-A and B
What did adding the barley HvNAS1 gene into rice do to rice grain iron content?
2-3 fold increase
What does increasing the expression of YSL2 gene in rice using a promoter do for Fe content?
Increased Fe content of grain
4 fold
What is ferritin?
Fe storage protein
Prevents anaemia and iron deficiency
What does adding a glutenin promoter that increases ferritin expression in rice achieve?
Ferritin expressed in seed endosperm and subaleurone
2-3 fold increase in Fe content
Why is diet bioavailability of plant nutrients such a problem
Different chemical forms, binding to different organic or non organic forms
- may inhibit transfer across membranes
Can be as low as 5% for most plant sources of iron
Can vary due to presence of ‘antinutrients’ - compounds that reduce absorption
What is phytates effect on the bioavailability of many essential nutrients?
Negative charge binds positive ions and interferes with assimilation
- considered an “antinutrient”
- binds minerals
- forms insoluble complexes that are not easily absorbed by the body.
How does phytase make some minerals more bioavailable in plants?
Break down metal-phytate complexes
- Release positively charged free metals (such as calcium, zinc, iron and mg)
- Increasing bioavailability of these minerals
What is phytate?
Storage form of phosphorus
What are Fer-NAS-YSL2 plants?
Plants with a combination of GM genes which increase iron content in their edible tissues
What are some potential negatives and challenges of Fer-NAS-YSL2 rice plants?
- risk of increasing accumulation of toxic metals (eg Cd)
- Expression worse in certain rice species
- More extensive field trials required - test yield
- Human feeding trials needed
What are GM strategies for Fe biofortification in rice grains?
Expression of nicotianamine synthesis enzymes
Expression of Fe-NA (YSL2) transporter
Expression of ferritin
What does expression of nicotianamine synthesis enzymes for Fe content in rice?
Improved long distance transport
How does expression of Fe-NA (YSL2) transporter increase Fe content in rice?
Increases transfer into rice grain
How does expression of ferritin increase Fe content in rice?
Improved Fe storage in grain
Why is high soil salinity such a big problem?
Reduces soil water potential
Effects large amounts of farmland
Reduces yield
Describe cellular dehydration caused by freezing?
Normal conditions - cell surrounded by water containing solutes
->
Ice formation begins
- Formation of ice decreases water potential outside the cell
->
Ice formation continues
- Higher solute concentration outside the cell draws water out causing dehydration and cell death
A plants water status is defined by _______ ______ measured in ____
Water potential
MPa
Water potential = _______ + ________
Water potential = solute potential + pressure potential
What is solute potential (osmotic pressure)?
Concentration of solutes dissolved in water
Water potential _________ as solute concentration increases
Water diffuses across semi-permeable membranes from ____ to _____ solute concentrations (osmosis)
low to high
What is pressure potential (turgor pressure)?
Physical forces exerted by water on cell membrane and cell wall
What occurs to cell turgor when water status is high?
When water pressure potential is high (high water status) water enters the cell causing the cell to become turgid
What occurs to cell turgor when water status is low (drought - water loss)?
Water leaves cells
- turgor is lost
- cells become flaccid
- Plant wilts
What are the short term impacts of water deficit?
Reversible effects
Leaf wilting
Declined photosynthetic rates
- stomata closed to reduce water loss
- reduced CO2 uptake
Reduced cellular/metabolic activity
- as energy from photosynthesis is required
What are the long term effects of water deficit?
- Not reversible
- Leaf abscission
- Cellular toxicity - Ion cytotoxicity
- Due to loss of water = < concentration of cellular contents
- Severely inhibited photosynthesis and respiration
What part of plants is very desiccation tolerant?
Seeds
Viable seeds can be recovered even after long term water deficit
What are the effects of high salt?
Altered Na+:K+ ratios
Increased NA+ and Cl-
Na+ rapidly enters root cell
Causing negative membrane potential
During periods of high salt concentration in soil describe the mechanism by which Na+ enters the root cell and what are the consequences?
Via non-selective channels and K+ transporters
- Outcompetes K+ for uptake
Na+ cytotoxic
- Many enzymes sensitive
- denature
K+ essential for plant function
What are the strategies focused in GM when developing crops with drought tolerance?
Osmoprotectants - Metabolites and molecules involved in protecting cell components from water stress effects
- Stress-responsive transcriptional regulation
- Osmoregulation via ion/water transporters
- Water use efficient carbon fixation
Describe xerophytes morphological adaptations
Xerophytes
- Thickened cuticles
- Reduce water loss
- Reduce leaf breakage from wilting
- Shiny cuticles
- Reflect light and heat
- Trichomes (leaf hairs)
- Trap damp air above stomata
- Reflect light
- Sunken stomata
- Small/no leaves
- Reduces surface area and water loss
Describe succulents morphological adaptations?
- Increased storage ability of water
Specialised organs
- Enlarged cells
- High volume vacuoles
Organ can be roots, stems, leaves
Low transpiration
Thick cuticles
How do drought resistant plants alter their metabolism to increase survival?
Modified photosynthesis and carbon fixation
Such as CAM and the C4 photosynthetic pathways
What does CAM photosynthesis achieve?
Improve water use efficiency
What are osmoprotectants?
Protect against osmotic stress induced by water deficit
- abundant in stress-tolerant plants and microbes
- non-toxic compounds
- do not disrupt hydration shell around the cell
What genes over-expression improve drought tolerance in plants? Producing succulence features - eg enlarged cells
CEB1
Why is the expression of CEB1 problematic in non-stress conditions?
Gives stunted growth
Energy intensive production - high metabolic demand
What does trehalose do in plants?
Confers stress tolerance
What enzymes produce trehalose?
TPS and TTP enzymes
How do plants control the expression of TPS and TPP? Give an example
Using inducible promoters to control expression
Rice rbcS promoter
- Specific to chloroplasts of leaf mesophyll cells
- Allows expression of TPSP (TPS and TPP fused together)
- only in chloroplasts of certain cells
- limiting energy demand through synthesis of trehalose
What is a water deficit specific control of trehalose production?
ABA-inducible promoter
- activated during water deficit stress
- ensures energy-intensive production of trehalose is only triggered when the plant is under stress
What is the functions of molecular osmoprotectants?
- Small organic molecules protect cells from osmotic stress (high salt concs)
Functions - Molecular chaperones = stabilise and protect conformation of proteins, RNAs and cell membranes
What are the two types of molecular osmoprotectants?
Proteins
- Heat shock proteins – HSPs
- Cold shock proteins – CSPs
What is an example of an example of a shock protein molecular osmoprotectant?
CspB - RNA chaperone from Bacillus subtilis
Expression of CspB in various plants has shown increased tolerance to cold treatment, heat treatment and drought treatment
What is DroughGuard?
Commercialised CspB transgenic maize in 2010
What is abscisic acid (ABA)?
Hormone abscisic acid (ABA)
- Stress hormone
- Synthesised in response to environmental stress
- Particularly water deficit
- Drought, salinity, freezing, cold
What are some downstream responses produced by ABA?
Osmoprotectants
Molecular chaperones/stabilisers
- Heat and cold shock proteins
Movement of water and ions
- Aquaporins, ion channels
Oxidative stress responses
What is the oxidative stress response in plants?
Peroxidase, superoxide dismutase
- Reactive oxygen species accumulate and lead to cellular damage
- Enzymes that scavenge ROS control indirect stresses
What are some osmoprotectants?
Sugars, proline, glycine betaine
There are _____ response pathways to drought stress, _____ ABA dependant and _____ ABA independent pathways?
There are 3 response pathways to drought stress, 2 ABA dependant and 1 ABA independent pathways?
Describe response to stress - ABA dependent or independent?
- Signal transduction ABA or ABA-independent
- in gene promoter regions
- Activate gene expression for stress response and tolerance
- Increasing transcription factor abundance may increase size of the response - enhancing drought resistance