Bioactive Natural Products natural products of ecological importance, produced by organisms (animals, plants, microorganisms) and mediating communication

Bioactive Natural Bioactive Natural ProductsProducts natural products of ecological importance, natural products of ecological importance,

produced by organisms (animals, plants, produced by organisms (animals, plants, microorganisms) and mediating communication microorganisms) and mediating communication between and among these organisms (between and among these organisms (chemical chemical ecologyecology))

most natural products have more functions (effects)most natural products have more functions (effects) parsimony (Nature saves energy)parsimony (Nature saves energy) dichotomy of effects (two or more functions of some dichotomy of effects (two or more functions of some

natural products)natural products) different organisms use the same compound(s) for different organisms use the same compound(s) for

different functions, in different contextdifferent functions, in different context biodiversity and chemodiversitybiodiversity and chemodiversity

22

IntroductionIntroduction chemical ecology - definition semiochemicals - pheromones, kairomones, defence

substances classification, structural types, examples practical use of pheromones

PheromonesPheromones

pheromones of social insects, organisation within a colony

pheromones of mammals pheromones of Homo sapiens

33

plant metabolites as a model for synthetic insecticides, repellents, and chemosterilants

regulators of growth, development, and reproduction of insects, insect hormones

juvenoids, juvenogenes, antijuvenoids, ecdysteroids

Biorational pesticidesBiorational pesticides

plant induced defence - „SOS“ signals allelopathy, natural and synthetic herbicides phytoalexins and pathotoxins natural and synthetic fungicides

Induced defence of plantsInduced defence of plants

44

Methods in chemoecologyMethods in chemoecology sample preparation (isolation) in chemical ecology identification and structure elucidation testing biological activity of natural products

chemistry of pollination, flower colours and fragrances food attractants (fagostimulants)

and deterrents, oviposition stimulants

Pollination and feedingPollination and feeding

55

EcologyEcologyrelations among organisms or groups of organisms and their environment

EEnvironmental sciencenvironmental sciencepollution, factors influencing environment in a negative way

Chemical ecologyChemical ecologychemically mediated interactions between organisms,communication between organisms

TypTypes ofes of intera interactionsctions: : insect - insect (pheromones, allomones, kairomones)insect - plant (host plant attractants, flower fragrance)plant - plant ("SOS signals")interactions between microorganismstritrophic and multitrophic interactions (plant-insect-insect; plant-insect-plant; plant-insect-pathogen)

66

Research in chemical Research in chemical ecologyecology

close cooperation of chemists and biologistsclose cooperation of chemists and biologists good knowledge of the biology of organism good knowledge of the biology of organism

whose signals are to be studiedwhose signals are to be studied prepared bioassay (how the identified prepared bioassay (how the identified

compounds will be tested)compounds will be tested) isolaisolation of activetion of active compoundscompounds chemicchemicalal anal analysisysis structure determinationstructure determination synthesis of active compoundssynthesis of active compounds bioassay bioassay – – in the laboratory (behaviour)in the laboratory (behaviour) in the fieldin the field

77

CommunicationCommunication exchange of information between exchange of information between

and among organismsand among organisms mediated by a set of signs or mediated by a set of signs or

signals shared by both partners, signals shared by both partners, the releaser the releaser and the receiverand the receiver

88

CommunicationCommunication means of means of animalsanimalsopticopticalal

movementsmovements, , mimesismimesis, , colourcolour

aacocousticusticbird’s singing and drummingbird’s singing and drumming, ,

insect insect stridulastridulationtion

99

chemicchemicalalsesecretions ofcretions of exo exoccrinrinee glandsglands

contactcontactccontaontactct sign signalsals in matingin mating, ,

daily hygienedaily hygiene

OH

OH

O

O O

HOH

H

CommunicationCommunication means of means of animalsanimals

1010

electricalelectricalin some fish familiesin some fish families

CommunicationCommunication means of means of animalsanimals

1111

AllAll organism organismss releaserelease chemic chemicalal signsignalsals a andnd rea reactct onon odoursodours of of

othersothers chemicchemicalal sign signalsals deliver life-deliver life-

important important informainformation quicklytion quickly,,simply,simply, unambiguouslyunambiguously

lower importance in humanslower importance in humans

the youngest discipline the youngest discipline in plantsin plants

1212

Insect’s senses of smell and Insect’s senses of smell and tastetaste

search for food (prey)search for food (prey) search for a partner, choice, matingsearch for a partner, choice, mating choice of a spot/host for egglayingchoice of a spot/host for egglaying aggregation (before winter or to overcome aggregation (before winter or to overcome

resistance of a host)resistance of a host) regulation of space and sufficient foodregulation of space and sufficient food alarm, defence, or attackalarm, defence, or attack organisation of social lifeorganisation of social life

smellsmell – transmitted by air (volatile compounds)tastetaste – contact (non-volatiles, water-soluble)

1313

SemiochemicalsSemiochemicals(chemical speech, information transfer)

Pheromonespherein (to carry) horman (excite)

(Karlson a Lüscher, 1959)

within a specieswithin a species

Pheromonespherein (to carry) horman (excite)

(Karlson a Lüscher, 1959)

within a specieswithin a species

Allelochemicalskairomones, allomones

synomonesbetween speciesbetween species

Allelochemicalskairomones, allomones

synomonesbetween speciesbetween species

cellscells (immune response) (immune response) insectsinsects (sexual (sexual behaviour, behaviour, regulation of regulation of social life) social life)

bacteriabacteria (chemotaxis) (chemotaxis) vertebratesvertebrates (dominance, (dominance, territory marking) territory marking)

algaealgae (attraction of gametes) (attraction of gametes) humanshumans (immunity, (immunity, sexual sexual behaviour) behaviour)

plantsplants (attraction of (attraction of pollinators)pollinators)

cellscells (immune response) (immune response) insectsinsects (sexual (sexual behaviour, behaviour, regulation of regulation of social life) social life)

bacteriabacteria (chemotaxis) (chemotaxis) vertebratesvertebrates (dominance, (dominance, territory marking) territory marking)

algaealgae (attraction of gametes) (attraction of gametes) humanshumans (immunity, (immunity, sexual sexual behaviour) behaviour)

plantsplants (attraction of (attraction of pollinators)pollinators)

1414

Sex Aggregation Trail Alarm Marking (space) Identification (social) Dispersion

Types of pheromones Types of pheromones according to biological according to biological

functionfunction

1515

Sex pheromones –Sex pheromones – attraction of mates, stimulation of copulation behaviour.

Aggregation pheromone –Aggregation pheromone – attracts more individuals of the same species. Purpose: aggregation for overwintering, gathering at food source, or mass attack and overcome the host resistance (bark beetles).

Trail pheromones –Trail pheromones – used by social insects to mark trails to food sources.

AlarmAlarm pheromones –pheromones – used by social insects in case of attack by predators.

Marking pheromonesMarking pheromones –– host etiketation to avoid multiple parasitizing, or territory marking.

Identification pheromones distinguish separate colonies of social insects (“home odour”).

Dispersion pheromones are a signal for running in different directions.

Types of pheromonesTypes of pheromones

1616

Sex pheromoneSex pheromone – attracts the opposite sex for mating

1717

SexuSexualal dimorfism dimorfism in mothsin moths, , different shape of antennaedifferent shape of antennae

gipsy mothgipsy moth,,malemale ♀ ♂♂♂ ♀

1818

Prove of existence and function of sex pheromoneProve of existence and function of sex pheromone

1919

Sex pheromones –Sex pheromones – attraction of mates, stimulation of copulation behaviour

Aggregation pheromone –Aggregation pheromone – attracts more individuals of the same species. Purpose: aggregation for overwintering, gathering at food source, or mass attack and overcomethe host resistance.







2020

Aggregation pheromoneAggregation pheromone – attracts individuals of both sexes at a long distance (bark beetles)

2121









2222

Trail pheromoneTrail pheromone – marks a trail to food sources (social insects - ants, termites…)

sisters

sisters!

2323









2424

Alarm pheromoneAlarm pheromone - releases defence behaviour

in other colony members, specifically soldiers (mostly in social insects - ants, termites, but also in species living socially in some periods

of their life – aphids, bugs)

2525

Prove of Prove of existencexistencee a andnd fun function of alarm pheromonection of alarm pheromone((termite soldierstermite soldiers))

2626









2727

Marking (space) pheromoneMarking (space) pheromone – marks an occupied territory (space), assuring enough food

for the next generation

cherry fruit fly

2828






IdentifiIdentificationcation pheromonespheromones distinguish separate colonies of social insects (“home odour”).

DispersDispersionion pheromonespheromones are a signal for running in different directions.


2929

pheromonespheromonesintraspecific communication

allelochemiallelochemicalscalsinterspecific communication

sexaggregationalarmmarkingidentificationdispersion

allomonallomonesesadvantage (profit) for the releaserkairomonkairomonesesadvantage (profit) for the receiversynomonsynomonesesadvantage for both

semiochemisemiochemicalscals

3030

ALLOMONEchemical signal advantageous for producer (releaser)

Spider Spider Mastophora cornigera

produces a mixture of Z9-14:Ald, Z11-16:Ald, Z9-14:Ac, and Z11-16:Ac

predator of ~ 15 moth species that use these compounds as sex pheromone (attracts males)

3131

Chemical defence(common, well studied)

Ladybird pupaLadybird pupa Epilachna borealiscovered by hollow hairs that secrete a sticky exudate containing a “combinatorial” mixture of alkaloids

As s result, predators (birds, ants) avoid the bad-tasting pupa.

Schröder, Science 1998

3232

KAIROMONzpráva výhodná pro příjemce

KAIROMONEchemical signal advantageous for receiver

Parasitic waspParasitic wasp Microplitis croceipes CaterpillarCaterpillar Spodoptera exigua

CaterpillarCaterpillar produces chemicals that make it “visible”

wasp localises the caterpillar based on chemicals from its faeces

parasitoids of insect eggs – a similar principle

3333

aphidsaphids x ladybirdsladybirds

aphids produce alarm pheromone (E-β-farnesene) that attracts predators, ladybirds

CH3 CH3 CH2

H3C

(E)- -Farnesene

CH2

KAIROMONEchemical signal advantageous for receiver

3434

SYNOMONEchemical signal advantageous for both sides

typictypicalal exampleexample – – flowerflower fragrancefragrance

plant offers nectar in flowers, insects pollinate its flowers

BUT:

orchids (Ophrys) attract males of solitary bees by a mixture of compounds resembling the sex pheromone of con-specific females, but males are not awarded with nectar (pollination by deceit)

3535

HistorHistory ofy of pheromonespheromones 19141914 Fabre J.H.Fabre J.H. males of moths are able to localise males of moths are able to localise

females for a long distancefemales for a long distance 1925-19391925-1939 biologicbiologicalal evidence of species-evidence of species-

specifity specifity of female attractantsof female attractants 19391939 Butenant A.Butenant A. first attempt to isolate attractant first attempt to isolate attractant

of the silk mothof the silk moth 1950-19611950-1961 isolaisolationtion a andnd identifi identification ofcation of

bombybombyccolol 19611961 synthesis ofsynthesis of allall 4 4 possiblepossible isomer isomerss 19591959 Karlson+LKarlson+Lüüscher scher introduced the termintroduced the term

„„phpheromoneromonee““ (Nature 183, 55)(Nature 183, 55)

silk moth

3636

IsolaIsolation of first sex tion of first sex pheromonepheromone

101066 pupae of silk mothpupae of silk moth glands of glands of 5x105x1055 femalesfemales dissecteddissected obtainedobtained 280 g 280 g of of lipidic extralipidic extracctt neutrneutralal frac fractiontion 125 g 125 g repeatedrepeated extra extractionction, , transesterificationtransesterification, ,

precipitation,precipitation, aandnd re reccrystarystalllilisationsation gave a crude fraction of gave a crude fraction of 44’-’-nitrobenzen-4-carboxylic acid esters (nitrobenzen-4-carboxylic acid esters (9 9 stepssteps)),, obtainedobtained 5,6 g 5,6 g

more separation stepsmore separation steps andand re reccrystarystalllilisationsation led toled to 6 6 mg mg of of nitroazobenzennitroazobenzenccarboxylarboxylate of ate of bomby bombykkolol

spespecctrtralal anal analysisysis (IR, UV) a (IR, UV) andnd derivati derivatissaation led to tion led to the structure determinationthe structure determination

synthesis of all synthesis of all 4 isomer4 isomerss

OH

bombykol

3737

PhPheromoneromones of mothses of moths MoleMolecularcular weightweight 100-300 100-300 represents a suitable represents a suitable

volatility, but also a sufficient number of structural volatility, but also a sufficient number of structural variants variants (10-18 (10-18 carbon atomscarbon atoms))

Most common types of compoundsMost common types of compounds – ali – aliphphatic atic alalccoholoholss, ester, esterss, aldehyd, aldehydeses, keton, ketoneses, , acidsacids

Less commonLess common – epoxid – epoxideses, ketal, ketalss, acetal, acetalss, , phphenolenolss High species specificityHigh species specificity – – differentdifferent chain length,chain length,

number of double bonds,number of double bonds, regioisomer regioisomerss, , stereoisomerstereoisomerss; ; more components in specific ratiomore components in specific ratio

Related species usually have similar pheromones Related species usually have similar pheromones ((similar biosynthetic pathwayssimilar biosynthetic pathways))

3838

Chemistry in the life of the Chemistry in the life of the rattlebox moth,rattlebox moth, Utetheisa Utetheisa

ornatrixornatrixT. Eisner T. Eisner && J. Meinwald, Cornell University, J. Meinwald, Cornell University,

USAUSA

adult

mating larvaon the host plant

Crotalaria spectabilis

defence

3939

Female pheromone of U. ornatrix

components of female pheromone

Female calls at the sunset, emitting a pheromone in 1,5 sec pulses.

Attracted male touches the female with his abdominal hairs (coremata). Acceptance results in mating.

courship

male coremata

4040

Male pheromone of U. ornatrix

N

CHOHO

(7R)-hydroxydanaidal(male pheromone)

monocrotaline

pheromone has aphrodisiacal effect on females

monocrotaline is transferred to female during mating, she transfers it further to her offspring (chemical protection/defence)

Female mates with several males. She determines male’s defensive vigour (the alkaloid content) and alkaloid donating capacity and chooses the sperm of the best male.

OO CH2O

N

H

OH

OH

O

Crotalaria mucronata

4141

Types of compounds in Types of compounds in pheromones pheromones

of different animalsof different animalsMoths (Lepidoptera)Moths (Lepidoptera)

OAc

OAc

OAc

OH

OAc

CHO

peach twig borerAnarsia lineatella

European corn borerOstrinia nubilalis

European corn borerOstrinia nubilalis

codling mothCydia pomonellaEuropean grapevine mothLobesia botrana

cotton bollwormHelicoverpa armigera

4242

Flies (Diptera; fruit fly = Flies (Diptera; fruit fly = vrtule)vrtule)

N

N

O

O

Papaya fruit fly

Mediterranean fruit fly

Caribbean fruit fly

4343

BeetlesBeetles (Coleoptera):(Coleoptera):bark beetles,bark beetles, weevilsweevils

H OH

OH

H

OH

OH

H

O

OO

H

O

(R)-ipsdienol (R)-sulcatol

(S)-cis-verbenol (1R,2S)-grandisol

chalcogran lineatin

Ips pini Gnathotrichus sulcatus

Ips typographus

Anthonomus grandis

Pityogenes chalcographus

Trypodendron lineatum

4444

Algae and seaweedAlgae and seaweed

HO OH

OH

O

COOH

OHOHO

HO

sirenin, Allomyces

lurlene, Chlamydomonas eugametos

green algae, males gametes

seaweed

4545

VertebratesVertebratesmuskmusk, , territory markingterritory marking

O

H

O

O

O

O

O

O

MusconMusconee, , (–)-((–)-(RR)-3-methylcy)-3-methylcycclopentadelopentadeccanonanonee

ExaltonExaltonee, , cycycclopentadelopentadeccanonanonee

CivetonCivetonee, , ((ZZ)-cy)-cycclohexadec-9-en-1-onlohexadec-9-en-1-onee

AmbrettolidAmbrettolidee, , ((ZZ)-hexadec-7-en-16-olid)-hexadec-7-en-16-olidee

ExaltolidExaltolidee, 15-pentade, 15-pentadeccanolidanolidee

African civetteAfrican civette

Siberian musk Siberian musk deerdeer

Musk oxMusk ox

4646

HumansHumans,,steroidsteroid compoundscompounds

HOH

H

vomeroferine

4747

Structure similaritiesStructure similarities

O

O

O

O

O

O

O

O

mouse, Mus domesticus

bark beetles, Dendroctonus spp.

(R,R)-exo-brevicomine (S,S)-exo-brevicomine

(R,R)-3,4-dehydro-exo-brevicomine

(S,S)-3,4-dehydro-exo-brevicomine

4848

ParsimonParsimonyy(dichotom(dichotomy of effectsy of effects))

(Z)-9-dodecenyl(Z)-9-dodecenyl acetacetateate

Female sex pheromone of many moth species and the female sex signal of African elephant.

O

O

4949

ParsimonParsimonyy ((thriftinessthriftiness, , economyeconomy))

O

HO

2,6-dimethylo2,6-dimethylocctanotanoicic acidacid

component of wax in some birds feathers impregnation disinfection (strong antibiotics)

5050

SSalicylalicylic acid and its ic acid and its derivativesderivatives

OH

O–-D-glucose

OR

OH

O

bark, leaves, and flowers of willow (Salix), bark and leaves of evergreen (non-deciduous) birch

salicinsalicineesalicylic acidsalicylic acid methylmethyl salicylsalicylateate

antipyretic effect in humans pyretic effect in flowers (heat production)

„SOS“ signalling in plants

5151

Defence substancesDefence substancesstructure examplesstructure examples

AcO

H

COOCH3

AcOH

H OH

HO

H

H

H

HOHOOH

H

O

HHO

H

H

H

HOH

H

H

neocembrene A

7,16-secotrinervitanetrinervitane

rippertane kempane 5-kempane

longipane

termite soldiers use chemical weapons (Nasutitermitinae)

5252

Other social insectsOther social insects

NO2

N

N

R NH

R NH

R1 R2

(E)-1-nitropentadec-1-enetermites, Prorhinotermes simplex

ants, families:Poneridae Myrmicidae

5353

Bombardier beetlesBombardier beetles ( (BrachynusBrachynus) ) OH

OH

+ H2O2

O

O

+ H2O

2 H2O2 2 H2O + O 2

peroxidáza

kataláza

5454

Other beetlesOther beetlesOH

O

O

HO

testosterone estrone

diving beetle, prothoracal gland

N

stenusinerove beetle Stenus comma

N

H

H H

O

coccinelineseven-spot ladybird,

Coccinella septempunctata

OO

O

O

cantharidinemeloid beetle (Meloidae)

5555

Structural curiosities as Structural curiosities as insect defence substancesinsect defence substances

OH

C

O

romallenonegrasshopper Romalea microptera

NH3, 4,5 %

burying beetle,Nicrophorus vespilio

HCN

millipedes,Chilopoda and Diplopoda

5656

Why to study Why to study pheromonespheromones??

many insect species are serious pests, many insect species are serious pests, causing great damage on cropscausing great damage on crops : :

cerealscereals fruits and vegetablesfruits and vegetables stored foodstored food ( (flourflour)) forests and woodforests and wood fabrics andfabrics and clothesclothes ( (woolwool, , fursfurs))

5757

InsecticidInsecticideses – – chemicals killing chemicals killing insectsinsects ( (non-selectivelynon-selectively))

Environment-friendly methods of Environment-friendly methods of plant protection are neededplant protection are needed

IIntegrated ntegrated PPest est CControl ontrol oror IIntegrated ntegrated PPest est MManagement)anagement)

5858

Disadvantages of the use Disadvantages of the use of insecticidesof insecticides

application of large amounts of application of large amounts of synthetic chemicals that stay in the synthetic chemicals that stay in the environment unchangedenvironment unchanged

toxicittoxicityy, , non-specific to target pestnon-specific to target pest repeated application is necessaryrepeated application is necessary development of resistancedevelopment of resistance killing of beneficial insectskilling of beneficial insects

5959

Advantage of the use Advantage of the use of pheromonesof pheromones

low concentrationslow concentrations specific to the target insectspecific to the target insect non-toxicnon-toxic

Disadvantage of the use Disadvantage of the use of pheromonesof pheromones

expensive productionexpensive production demands on formulation, traps demands on formulation, traps

checking, evaluation (qualified checking, evaluation (qualified management)management)

6060

Practical use of Practical use of pheromonespheromones

MMonitoringonitoring of the flight and population of the flight and population density of pest insect; pheromone trap density of pest insect; pheromone trap replaces a female (in moths) and lures replaces a female (in moths) and lures males; pheromonal dispensers and traps.males; pheromonal dispensers and traps.

MMass trappingass trapping – – use of aggregation use of aggregation pheromone in case of a high population pheromone in case of a high population densitydensity; ; lures both sexeslures both sexes ( (attract and killattract and kill).).

MMating disruptionating disruption – – “high” concentration “high” concentration of sex pheromoneof sex pheromone; ; males are unable to males are unable to localize a female and thus, no mating localize a female and thus, no mating occursoccurs..

6161

MMonitoringonitoring of the flight and population density of the flight and population density of insectsof insects; pheromone trap replaces a female (in pheromone trap replaces a female (in moths) and lures males; pheromonal dispensers moths) and lures males; pheromonal dispensers

and traps.and traps.

6262

pheromonal dispensers pheromonal dispensers pheromonal trapspheromonal traps

6363

MMass trappingass trapping – – use of aggregation use of aggregation pheromone in case pheromone in case of a high of a high population population densitydensity; ; lures both lures both sexessexes

Attract and killAttract and kill – – combination with combination with insecticideinsecticide

6464

MMating disruptionating disruption – – “high” concentration of “high” concentration of sex pheromonesex pheromone; ; males are unable to localize a males are unable to localize a female and thus, no mating occursfemale and thus, no mating occurs..

6565

World use ofWorld use of IPM IPM

Monitoring 32.1 %

Mass trapping 23.3 %

Attract-and-kill 2.2 %

Mating disruption

42.4 %

Data from nineties, 25 countries. Pheromone-treated areas represented 1 % of all fields.

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Bioactive Natural Products natural products of ecological importance, produced by organisms (animals, plants, microorganisms) and mediating communication