Strigolactone - A New group of Phytohormone

T.Parthasarathi,Ph.D.Scholar(CropPhysiology) DepartmentofCropPhysiology, TamilNaduAgriculturalUniversity,Coimbatore,India.

(Authoremailid:tparthasarathicrp@gmail.com)

Strigolactones (SLs)

SLs are a class of sesquiterpene lactones are derived from carotenoids & contain a large four ring backbone (Matusova et al., 2005)

Biosynthesis occurs mainly in roots & isolated from roots of a variety of plant species

Natural SLs isolated from plant roots are mainly 5-deoxy-strigol, strigol, sorgolactone, alectril & orobanchol

Synthetic analogues GR24, GR6 & GR7 - at pg level as natural strigolactones

Historyofstrigolactoneinven1on
Germination stimulant for Striga was isolated in 1966 from root exudate of cotton, which is not a host for Striga or Orobanche

Structure of strigol was elucidated in 1972, its absolute stereochemistry was established by X-ray diffraction analysis in 1985

Sorgolactone a compound with a structure similar to strigol was isolated from roots of sorghum (Mori 1998); CCDs - (Aldridge et al., 2005); Tillering in rice (Zhou et al., 2006) Collective name strigolactones was proposed for class of molecules

General structure & numbering scheme of Strigolactones

Conversion of 5-deoxystrigol to other strigolactones

5-Deoxystrigol: Common Precursor of various SLs

SLs Biosynthesis
3 enzymes identified that are involved sequentially in their synthesis 1. carotenoid- cleaving dioxygenase 7 (CCD7, MAX3/RMS5 /HTD1/D17) 2. carotenoid-cleaving dioxygenase 8(CCD8, MAX4/ RMS1/DAD1/D10) 3. cytochrome P450 monooxygenase (Cyt P450, MAX1) Enzymes solely responsible for synthesis of such complex SLs compounds Two independent pathways to synthesize sesquiterpenoids 1. SLs produced by plastidic methylerythritol phosphate (MEP) pathway 2. Independent by cytosolic mevalonic acid pathway

Biosynthetic pathway of Strigolactone

Schematic representation of biosynthesis of strigolactones

Black arrows - Carotenoid-derived secondary metabolites

Dotted arrows - possible interactions bw different components in the scheme

Open arrows - several conditions or strategies - modify the production of strigolactones (+ve effect on strigolactone production; ve effect on strigolactone production)

NCED,9cisepoxycarotenoiddioxygenase; CCD,carotenoidcleavagedioxygenase. Hormones:ABA,abscisicacid

MovementofStrigolactone

ccd8 mutant Orobanchylacetate (MW388.42,C21H24O7)showingthefourringstucture Endogenousstrigolactonesinpea, (GomezRoldanetal.,2008)

synthe1cstrigolactoneGR24 (MW298.3,C17H14O5) (Umeharaetal.,2008)

GR24intothestemofpeaplants Moveupward(Transpira1onstream)
(Gomez-Roldan et al., 2008)

Biological functions of strigolactones

Seed Germination Activity

Hyphal Branching Activity

Shoot-Branching Inhibition

Inhibition of bud out growth

Srigolactones & Host Specificity

(Xie et al., 2010)

Seed Germination Activity

Natural & synthetic SLs have moderate to potent germination activity Breaks secondary dormancy Hydrolysis of SLs structure in synthetic SL GR24 affording the ABC part & D-ring moiety resulted in a complete loss of germination activity 2-epiorobanchol is slightly more active than orobanchol on seed germination of O. minor & P. ramosa (Orobanche ramosa)

Germinating S. hermontica seeds on 0.1 mM GR24, 25 mM trans-zeatin & DMSO (Control)

SLsalleviatethermoinhibi1onofseedsofArabidopsis

GR24 ABA levels & GA levels during alleviation of thermoinhibition

(Toh et al., 2012)

ShootBranchingInhibi1on
SLs inhibit shoot branching by preventing bud outgrowth of leaf axillary buds

SLs require receptor in order to exert their activity - Receptors are active at very low conc & transported in plant over some distance Under Pi deficiency, plants increase the biosynthesis of SLs to reduce shoot branching Maximizes symbiotic interaction with AM fungi to facilitate the uptake of mineral nutrients Strigol, 5-deoxystrigol & GR24 were active in bioassays for shoot-branching inhibition

SLs - central role in coordinating shoot branching, AM symbiosis & parasitic weed seed germination

IAA, Indole-3-acetic acid; CK, cytokinin; St, strigolactone

SLs are produced in plant root, from which some is release to soil to induce hyphae branching & some is translocated to shoots to inhibit shoot branching Shoot branching - Inhibited by IAA from shoot & promoted by CK from root Biosynthesis of SLs is regulated by soil nutrient availability

Simplified model of shoot branching inhibition via strigolactones & MAX/RMS/D pathway

(Beveridge and Kyozuka 2010)

Strigolactone pathway for shoot branching inhibition

Mutant-induced branches on these plants are indicated by arrows

(Dun et al., 2009)

Gene1cvaria1oninSLproduc1on&1lleringinrice

Strigolactone conc in root exudates (a) root tissues (b) of rice cultivars

(Jamil et al., 2011)

Inhibi1onofbudoutgrowth
Schematic representation of inhibition of bud outgrowth by SLs

(Goulet and Klee 2010)

Strigolactones inhibit bud outgrowth Production requires - carotenoid cleavage dioxygenase-like enzymes CCD7 & CCD8

Step is regulated by auxin & an unknown long-distance feedback signal

(Taiz and Zeiger 2010)

Simplified model of bud outgrowth regulation by the SMS pathway exemplified in pea

(Beveridge 2006; Foo et al., 2007)

Hyphal Branching Activity

Mycorrhizal fungi perform a highly beneficial function Mycorrhizal fungi pass on water & nutrients (P), fungi absorb from the soil through their fibrillose hyphae Mycorrhizal fungi develop a symbiotic relationship with host plant & host plant actively secretes SL to attract beneficial mycorrhizal fungi Nutrient-poor environment, host plants attract mycorrhizal fungi, which help plants to absorb nutrients Same time, root need to prevent stem from unfruitful shoot branching Hyphal branching in AM fungus G. margarita & all natural SLs were found to be active as branching factors

Cont d
3,6-dihydro-GR24 was inactive as a germination stimulant but still showed distinct activity on hyphal branching (K. Akiyama, unpublished data) Strigolactones present in root exudates & inducing hyphal branching of AMF -important plant recognitions signals for AMF Branching factor(s) in root exudates - Regulating mechanism for root colonization - effect of root exudates from plants with low P-levels on hyphal branching Arabidopsis, rape, cabbage (Brassicacea) & sugar beet (Chenopodiaceae) are AM non-host plants - root exudates of these plants (Benharrat et al., 2003) not showed a hyphal branching activity on AMF - because lack of branching factor

Relationships among strigolactone, root-parasitic plants & mycorrhizal fungi

SLs, signals for parasitic plants & AM fungi

(Garca-Garrido et al., 2009)

SLsreleasedbyroots,inducehyphalbranchinginproximityto theroot

(Gmez-Roldn et al., 2008)

Strigolactones influence germination of parasitic seeds & branching patterns of mycorryzial fungi

Strigolactones and Host Specificity

Plants produce mixtures of SLs & release them into the rhizosphere In rhizosphere, less stable hydroxy- SLs (orobanchol & strigol) may decompose more rapidly than 5-deoxystrigol & sorgolactone Less stable SLs can induce seed germination & hyphal branching when the parasite seeds & AM fungi are in close proximity to the roots

SLs may Affect Shoot & Root Development

SLs application on growing buds reduced their growth, with no effect on auxin transport from bud (Brewer et al., 2009) SLs inhibit polar auxin transport from buds by reducing the capacity for transport from apical meristem, resulting in restrained bud outgrowth (Leyser 2009; Bennett et al., 2006; Ongaro and Leyser 2008) SLs serve as auxin-promoted secondary messengers that move up into the buds to repress their outgrowth SLs repress lateral root initiation GR24 treatment led to an increase in root hair length, which might be mediated via the MAX2 F-box (Kapulnik et al., 2010)

Strigolactones & Environmental Cues Strigolactones (SLs) - Role in plant developmental plasticity in roots & shoots in response to environmental cues
(Light & Nutrient status)

(Koltai and Kapulnik 2011)

Light status
Light quality & intensity affect multiple processes in plants, including determination of shoot architecture SLs +ve regulators of light associated processes Light quality, i.e., a low R:FR ratio suppress shoot branching (part of the shadeavoidance response) - SLs identified as shoot-branching inhibitors - one of the mediators of response (Franklin 2008; Leyser 2009; Franklin and Whitelam 2005 ) SLs may regulate shoot branching in response to light quality on one hand & light harvesting on other one SLs - suggested to be associated with inhibition of lateral root initiation & root hair elongation (Schlicht et al., 2008) Cross-talk bw SLs & light-associated pathways follow a feedback loop, because carotenoid biosynthesis is light-dependent & SLs are thought to be derived from this pathway (Matusova et al., 2005)

Nutrient status
(1) SLs inhibit shoot branching (2) SLs modulate auxin transport (auxin transport is involved in shoot s response to B deficiency (Wang et al., 2006) (3) SLs biosynthesis is responsive to nutrient (P [Pi] & N) levels (Yoneyama et al., 2007; Lpez-Rez and Bouwmeester 2008) SLs provide a way to coordinate shoot development with nutritional conditions Root hairs development affected by SLs (Kapulnik et al., 2010; Schlicht et al., 2008; Koltai et al., 2010) Low Pi & low N conditions induce SLs production (Lpez-Rez et al., 2008) SLs are mediators of the root response to low nutrient condition

Study results
SLs are suggested to increase the plant s light-harvesting ability Inhibit shoot branching; SLs induced increase in light-harvesting capability compensates for reduction in total LA resulting from reduced shoot branching Low levels of nutrients (Pi & N) increase root SLs production Levels of SLs & putative precursor lead to alterations in root-hair length Root hair length directly associated with enhancement of the plant s ability to absorb nutrients (Gilroy2000) SLs affect root-hair elongation, resulting in modulation of nutrient uptake by the plant

Signal Transduction
MAX2 and RMS4 are a putative signaling component of branching signal MAX2/RMS4/D3 encode F-box proteins which are typically involved in ubiquitin mediated protein degradation Level of epi-5-deoxystrigol (epi-5DS), a native strigolactone of rice, was elevated in d14 mutants & d14/htd2 mutants show feedback upregulation of SL biosynthesis pathway genes D14 func1ons downstream of SL synthesis determina1on of D14 func1on & iden1ca1on of poten1al nega1ve regulators that are degraded by the MAX2/ RMS4/D3 pathway are urgently required to reveal the percep1on & signaling mechanismsofthebranchinghormone

SLs on nodule formation in alfalfa (Medicago sativa)

Data represent mean number of nodules per plant (n = 20) Error bars represent standard errors at 95% confidence interval

Nodulation in Medicago sativa inoculated with Sinorhizobium meliloti after application of different GR24 solutions & corresponding control solutions Application of GR24 alter phytohormones levels (auxin & cytokinin) with crucial roles in nodule organogenesis (Arite et al., 2007; Hayward et al., 2009)
(Soto et al., 2010)

Overview on plants with strigolactone production

Plant species Hostfor Striga and/or Orobanche Striga Striga Striga Orobanche Orobanche Striga Striga/ Orobanche Orobanche Strigolact. detectedin root exudates Yes Yes Yes yes Yes yes yes Root exudates s1mulate Striga/Orob. germinat. Strig.+Orob. Strig.+Orob. Orob. Orob. Orob. Strig. Strig.+Orob. Orob. Root exudate ini1ate branching ofAMF Yes Yes Yes Yes yes yes Reference

Maize Sorghum Millet Pea Redclover Cowpea Tobacco Tomato

(Siameetal.,2003) (Haucketal.,1992) (Siameetal.,2003; Awadetal.,2006) (Rubialesetal.,2006) (Yokotaetal.,1998) (Mlleretal.,1992) (Siameetal.,2003; Gonzalezetal.,2005) (Yoneyamaetal.,2004)

Strigolactone regulation of shoot branching in chrysanthemum

Effect of GR24 & NAA on bud outgrowth on two-node isolated stem segments

Error bars represent the standard error of the mean, n=23-24

(Liang et al., 2010)

Transgenic approach for CCD8 transcript expression

DgCCD8 transcript response to decapitation, auxin & GR24

(Liang et al., 2010)

Cont d
GR24 - Potential use for regulation of branching in chrysanthemum horticultural production

Large-scale exogenous treatment techniques need to be developed & cost of GR24 or analogues needs to be reduced practical application Chrysanthemum CCD8 genes providing opportunity to use genetic modification to improve the chrysanthemum growth habit Effects of GR24 on bud activity & DgCCD8 expression support the idea that SLs inhibit bud activation by modulating auxin transport canalization Auxin is a major mediator of feedback by SL on SL synthesis

REGULATION OF STRIGOLACTONE PRODUCTION

Under nutrient starvation, plants increase SL production, minimize excessive shoot branching & promote AM colonization

Phosphate starvation - expression of LeNCED1 in tomato nor expression of D10 in rice were upregulated (S. Yamaguchi & K. Yoneyama, unpublished data)

Increase in SL production under nutrient starvation might be regulated post transcriptionally

Nodulating leguminous plants - SL production is promoted only under P deficiency, whereas in other mycotrophic plants it also occurs under N deficiency (K. Yoneyama, unpublished data) Nonmycotrophic plants (white lupin (L. albus) & spinach (Spinacea oleracea)) exude SLs at low levels as compared with mycotrophic plants, nutrient deficiencies hardly affect SL exudation (K. Yoneyama, unpublished data)

Inhibitors of SLs
Fluridone & Norflurazon herbicides inhibiting carotenoid biosynthesis (inhibitors of phytoene desaturase), could reduce SL production, inhibit later steps of SL biosynthetic pathway Hydroxamic acid - type CCD inhibitors affecting SL production This inhibitors induce a branching phenotype in Arabidopsis that is similar to that of ccd7/8 mutants carba-GR 24, carba-GR 7 & carba- GR 5

Practical application in agriculture

Natural SLs isolated from plant root exudates as germination stimulants, except for 5-deoxystrigol was identified as a branching factor, an essential signal for root colonisation by AM fungi SLs or their metabolites are novel class of plant hormones regulating plant aboveground architecture Novel tools or approaches for parasitic plant management Approach for pruning chemical in larger horticultural crop fields Application of GR24 on legumes have crucial role on increasing nodulation better suitable for better nutrient use from soil (Arite et al., 2007; Hayward et al., 2009)

Cont d
SLs identified as stimulators of the germination of root-parasitic weeds that pose a serious threat to resource-limited agriculture SLs exuded from roots & function as signalling compounds in the initiation of AM, which are plant-fungus symbionts with a global effect on C & P cycling SLs established to be phytohormones that regulate plant shoot architecture by inhibiting the outgrowth of axillary buds Manage germination of parasitic weeds responsible for massive crop losses & control of lateral branching in forestry to produce higher quality woods Understanding SLs pathway has potential to significant impact in agriculture

Practical application in agriculture

Nijmegen-1 in suicidal germination approach - Orobanche infestations tobacco field Concentration of stimulant spray to soil was 7 x107 mol L1, corresponds to 6.25 g of stimulant ha1 - very small amount indeed for a herbicide

Tobacco field treated with Nijmegen-1

Untreated tobacco field

carba-GR, carba-GR 7 & carba- GR 5 - compounds accepted by protein receptor, block active site & germination of the seeds would be inhibited used in soil seed banks

Conclusion
SLs have three distinct biological activities: Induction of seed germination of root parasitic seeds, induction of hyphal branching of AM fungi & inhibition of shoot branching in plants SLs derived from carotenoids through sequential oxidative cleavage & subsequent oxidations Plants regulate production & exudation of SLs in response to changes in their environments Plants under nutrient starvation produce more SLs to minimize excessive shoot branching & to promote root colonization by AM fungi