Host Recognition Strategies of Striga - A Parasitic Angiosperm


Host Recognition Strategies of Striga - A Parasitic Angiosperm

William Stump
Colorado State University
Fort Collins, Colorado 80523

Abstract.  Parasitic angiosperms are unique life forms that are 
parasitic to other higher plants.  Many of these parasitic plants have 
developed complicated host recognition strategies.  This review will 
focus on Striga, a genus of the Scrophulariaceae, that contains many 
parasitic taxa of agronomic importance.  Striga species are root 
parasites and most are obligate parasites.  Many Striga species have 
narrow host ranges as a result of co-evolution with host plants in the 
agroecosystem.  Specificity is ensured by chemical signals from the host 
plant.  In addition to normal germination requirements common in many 
angiosperms, Striga species require a chemical stimulation from the host 
before germination can occur.  These chemical cues have been identified 
as strigol, and other analogs of this  sesquiterpene.  Also, a variety 
of other artificial chemical compounds have been found to initiate 
germination.  Strigol is thought to induce germination via ethylene 
biosynthesis.  For haustorial initiation an additional unique chemical 
signal is required.

INTRODUCTION

   Parasitic angiosperms are unique life forms that are parasitic to 
other plants.  These plants have evolved independently in 20 plant 
families, comprising some 3000-5000 species (Musselman, 1987).  
Parasitic plant classification is based on several criteria. 
Holoparasites lack chlorophyll and depend entirely on the host for 
nutrients, minerals and water; examples include Cuscuta sp. and 
Orobanche sp.   Hemiparasites have chlorophyll and thus can fix CO2 
depending on the host for water and minerals; examples here include 
Striga sp. and mistletoes.  Some parasites attack the shoot, others the 
root.  Parasites that require attachment to the host to complete their 
life cycle are obligate parasites whereas facultative parasites can 
complete their life cycle without the host.  Despite parasitic plant 
diversity, their common feature is the special host contact organ the 
haustoria.  With the haustoria, the parasitic plant is able to penetrate 
the hosts epidermis and establish a connection.
  Parasitic plants are a heterogeneous group being represented by 
generalists, with a wide host range, and specialists with narrow host 
ranges depending on their chosen co-evolutionary pathway (Musselman, 
1987). 
  Though the number of parasitic genera is great, the bulk of the 
research has been concentrated on agronomically important genera from 
five families that include the following:

	Scrophulariaceae: Striga
	Orobanchaceae: Orobanche
	Cuscutaceae: Cuscuta
	Visacaceae/Loranthaceae: Mistletoes

   Seedlings of obligate parasites, in order to survive, must quickly 
find a suitable host or they will soon die.  To accomplish this, 
parasitic life forms have evolved complicated host recognition 
strategies.  These strategies include host chemical cues for germination 
and haustoria formation, responses to light environment, and reliance on 
avian seed predators to disseminate seeds to appropriate hosts 
(Musselman, 1980; Calder and Berhardt, 1983).  This review will focus on 
Striga, a genus that contains many agronomically important weeds.  This 
genus causes considerable yield losses in various crops in tropical and 
subtropical countries.  The majority of Striga species are root 
hemiparasites and most are obligate parasites.  The most studied are S. 
asiatica (L.), S. hermonthica (Del.) pests of graminous crops, and S. 
gesneriodes (Willd.) which parasitize legumes (Musselman, 1980; Parker 
1991).  Many Striga species have narrow host ranges as a result of co-
evolution in the agroecosystem.  Host specificity is ensured by chemical 
signals from the host plant.  

Striga Life Cycle

   The life cycle of Striga is composed of five stages, as follows:

1. Germination (after seed preconditioning)
2. Haustorial initiation
3. Penetration of host tissue
4. Physiological compatibility
5. Parasite growth and maturation preconditioning

   Prior to germination Striga must undergo an after-ripening period 
before it will respond to any germination stimulants.  This after-
ripening is analogous to other angiosperm seeds that possess an innate 
dormancy.  For most of the species studied, seeds require certain 
temperature and moisture regime of a period of about two weeks.  It is 
thought that this period is necessary to remove phenolic compounds that 
act as germination inhibitors (Musselman, 1980).  Subsequent to the 
after-ripening, seeds of Striga require a period of conditioning before 
responding to germination stimulants.  Typically, warm moist conditions 
are required for about a week before maximum germination will take place 
after receiving the host stimulatory cue.


The Germination Process  

   The seeds of Striga are very small, measuring 0.15 x 0.31 mm, so it 
is quite obvious that Striga seeds lack the reserves for sustained 
periods of growth before host attachment.  For successful host 
attachment germination must take place within 3 to 4 mm of the host root 
since Striga radicles can manage only 2-4 mm (Ramaiah et al., 1991).  To 
compensate for these biological restrictions Striga produces up to 
450,000 seeds per plant, with a persistence in the soil of up to 10 
years (Eplee, 1992).  The most important, and evolutionarily complicated 
adaptation, is Striga's requirement of a germination stimulant from a 
nearby host.  

Germination Stimulation

   Many chemicals have been discovered that promote germination in 
Striga.  These chemicals include ones that are naturally occurring in 
plant root exudates, various synthetic plant hormones, and a variety of 
other derived compounds.  Because of potential control benefits, many 
researchers have attempted to isolate and identify natural host 
stimulants.  It has been found that host plants produce a mixture of 
stimulatory compounds with one compound being predominant over the 
others (Worsham, 1987; Weerasuriya et al., 1993).  These compounds have 
been found to be very active and are present in root exudates at 
extremely low levels.  This potent behavior has led some researchers to 
speculate that these germination stimulators represent a new class of 
plant hormones (Cook et al., 1972; Siame et al., 1993).  Unfortunately, 
the low concentrations and instability of these compounds has made 
isolation and identification difficult.
    The first naturally occurring germination stimulant characterized 
was strigol, a sesquiterpene isolated from cotton, a non host plant 
(Cook et al., 1972).  Strigol has activity in the soil solution at 
concentrations as low as   10-15  mol m -3.  The molecule, whose 
absolute structure was established in 1985 (Brooks et al., 1985), 
consists of four rings:  a six-carbon ring (A), a five-carbon ring (B), 
a four-carbon lactone ring (C) coupled to another lactone ring (D) by a 
=C-O- connecting unit. (see figure 1).  Once an active structure was 
established, many strigol analogs have been synthesized along this ABCD 
ring structure, and tested for activity.  These compounds were termed 
"GR" compounds and several had fairly good activity (Worsham, 1987).









Work by Mangnus and Zwannengurg (1992) along these lines has shed some 
light on the molecular activity of these compounds.  The research 
suggests the importance of the linkage between the two lactones is 
crucial for activity.

Identification of Host Germination Stimulants

   Many attempts have been made to isolate Striga germination stimulants 
from true host plants.  The first host derived stimulant was sorgoleone 
from Sorghum bicolor hydrophobic root exudates (Chang et al., 1986; 
Netzly et al., 1988).  The compound is an unstable dihydroquinone which 
is rapidly oxidized to a stable inactive quinone (see figure 2).  Unlike 
strigol, sorgoleone is hydrophobic and not as active with activity down 
to 10-7 M.  Chang et al. (1986) hypothesized that the ephemeral nature 
and limited mobility of the compound was capitalized by Striga to ensure 
germination in close proximity to the host root.
   However, in 1991 an additional new stimulant, sorgolactone, was 
isolated from water soluble sorghum root extracts (Hauck et al., 1991).  
This new compound is very similar to strigol in structure, solubility, 
and activity on Striga germination (figure 2)








     Figure 2. Structures of (a) strigol, (b) sogolactone, and (c) 
sorgoleone..
   Recent work done on characterizing resistant and susceptible sorghum 
cultivars on agar gel assay (Hess et al., 1992) and a dilution assay 
(Weersuriya et al., 1993) found great differences in the amount of 
germination stimulators for Striga asiatica present in sorghum root 
extracts.  The resistant cultivars, where resistance is based on low 
stimulant production, produced the same amount of sorgoleone as 
susceptible cultivars.  These findings cast a shadow on the importance 
of sorgoleones as Striga stimulants.  It was concluded that since the 
agar gel assays measured more water soluble stimulants and correlated 
well with strigol, that the main germination stimulant was the more 
water soluble sogolactone rather than sorgoleone (Hess et al., 1992).
   An additional Striga stimulator was found in cow peas, Vigna 
unguiculata, which is parasitized by Striga gesnerioides  a species that 
has evolved to parasitizes mainly legumes.  Legumes are frequently used 
as trap crops for Striga sp. that parasitize cereals since they are non 
hosts.  Cow peas, produce a variety of Striga stimulators.  The most 
prevalent compound was identified as alectrol, a strigol analog (figure 
3)





Figure 3. Alectrol (proposed structure)
.
   It was not until 1993 that strigol was isolated from the Striga host 
plants maize (Zea mays L.) and proso millet (Panicum miliaceum L.) by 
Siame and others (1993).  Very small amounts of strigol was also found 
in sorghum but the major stimulant was thought to be that of 
sorgolactone.  The authors felt that identification of strigol in true 
host plants took so long because most studies utilized sorghum root 
extracts of which strigol is only a minor component.  Also, strigol is 
present in very low concentrations in host root extracts making 
isolation difficult.

Other Chemicals that Stimulate Striga Germination

   In addition to natural host exudates and analogs of strigol a number 
of unrelated and diverse chemicals have been found to stimulate Striga 
germination in vitro.  These include several growth hormones; kinetins, 
zeatin, and ethylene (Worshum, 1987).  The use of ethylene is the 
backbone of the US witchweed (Striga asiatica) eradication program.  
Ethylene injected into the soil has been found to promote germination up 
to 90% to a depth of 30 cm at a concentration of 10-1 µl/L (Eplee, 
1992).  Somewhat less efficacy of ethylene was found on the stimulation 
of Striga hermonthica in Africa due to the drier soil conditions (Bebawi 
and Eplee, 1986).  Other chemicals that induce germination include 
scopletin, inositol, and sodium hyperchlorite.  It is not clear at this 
time how these various chemicals can stimulate germination (Worshum, 
1987, Igbinnosa et al., 1992).
   
Mechanisms of Germination Stimulation in Striga 

   Mechanisms by which strigol and its analogs stimulate germination in 
Striga is just recently starting to unfold.  Much of the work has 
concentrated on the role of ethylene on germination.  Since ethylene 
promotes germination in parasitic plants such as Striga, as well as many 
other non parasitic angiosperms, it has been hypothesized that strigol 
stimulates germination by promoting ethylene biosynthesis (Logan and 
Stewart, 1991).  This could also possibly explain how the diverse 
artificial chemical stimulants could promote germination by indirectly 
initiating ethylene production via a wound response (Gabbar et al., 
1993).  Strigol stimulated ethylene biosynthesis in preconditioned 
Striga asiatica seeds within three hours of application and prior to 
radicle elongation (Gabbar et al., 1993).  Both germination and ethylene 
production were inhibited by aminoethoxyvinyl glycine (AVG), an 
inhibitor of ethylene biosynthesis (Logan and Stewart, 1991) and by 2,5-
norbornadiene, an inhibitor of ethylene action (Gabbar et al., 1993).  
Addition of an ethylene biosynthesis intermediate, 1-amino-cyclopropane-
1-carboxylic acid (ACC), was found to override the inhibitory effect of 
AVG (Gabbar et al., 1993).  Strigol is thought to trigger activation 
and/or synthesis of ethylene forming enzymes for subsequent conversion 
of ACC to ethylene (Gabbar et al., 1993).  Ethylene has been found to 
stimulate germination in a number of plant species and is thought to 
initiate the biochemical cascade leading to germination (Logan and 
Stewart, 1991).
   There has been great interest in using a stimulant as a method for 
control of striga in a cropping situation.  Introduction of a 
germinating agent before a crop is planted can reduce striga populations 
via suicide germination (Eplee, 1975).  Since strigol is such a 
complicated molecule, requiring 12 to 20 steps to synthesize, a sizable 
amount of work has gone into determining which parts of the molecule 
confer activity in the hopes of finding a simpler to synthesize compound 
(Vail et al., 1990; Mangnus et al., 1992).  Taking the ABCD ring 
structure of strigol, various analogs were made along the ABC portion of 
the ring.  It was found that the D portion of the molecule was necessary 
for activity in conjunction with A and or  B with C not as important for 
activity (Mangnus, et al., 1993).  Work has not been conclusive in this 
area and the absolute molecular mechanisms have yet to be established.

Haustoria Initiation

   Host recognition in parasitic angiosperms is typically presented at 
two levels, depending on the level of parasitism.  For obligate 
parasites such as Striga, where a parasitic relationship is required for 
completion of life cycle, host recognition is determined by stimulation 
of germination via a chemical cue from the host as discussed in the body 
of this paper.  For facultative parasites, where parasitism is not 
necessary to complete it's life cycle, host recognition is accomplished 
by stimulation of haustoria formation by a chemical cue from the host 
(Steffens et al., 1986).  The first haustoria inducers isolated for the 
facultative root parasite Agalinis purpurea were phenylpropanoids named 
xenogin A and B extracted from Astragalus gummifer a non host plant 
(Lynn et al., 1981)  However, since these compounds were extracted from 
aerial portions of Astragalus gummifer  it's biological relevance was in 
question.  Investigating exudates from Lespedeza sericea, a true host of 
Agalinis purpurea , a pentacyclic tritriterpenehaustorial named 
soyasapogenal was found to be a haustorial inducer (Steffens et al., 
1986).  However its chemistry is quite different from the 
phenylpropanoids and is not as active and may depend on other yet 
identified materials for full activity (Steffens et al., 1986).
   It has been argued that host recognition in obligate root parasites 
such as Striga is by chemical cues for both seed germination and 
haustoria formation (Musselman, 1980).  Indeed, the fact that nonhosts 
such as cotton and cowpea both can stimulate Striga germination but not 
allow for a parasitic connection illustrates the need for an additional 
chemical cue.  Striga asiatica, haustoria initiation is in response to a 
single quinone, 2,6-dimethoxy-p-benzoquinone (2,6-DMBQ) found in sorghum 
root extracts (Chang and Lynn, 1986).  However, 2,6-DMBQ is not found in 
root exudates, its presence requiring vigorous shaking of root tissue, 
which indicates its within the host root tissue (Chang and Lynn, 1986).  
A byproduct of lignin degradation by fungi with extracellular lactases 
is also 2,6-DMBQ.  Coupled with the fact that Striga utilizes cell wall 
degrading enzymes, it has been suggested that Striga lactase activity 
releases quinones from the host root that stimulate haustoria initiation 
(Stewart and Press, 1990).
  However, calling haustoria initiation in Striga a host recognition cue 
is somewhat misleading.  Once the parasite has germinated it had better 
be in the presence of a suitable host or it will soon die, further host 
selection at this juncture is pointless.  This is the basis of the 
control technique of "suicide germination".  I propose that this 
secondary chemical cue is strictly for the detection of the proximity of 
the host root.  A time course study of establishment of Striga asiatica 
on susceptible sorghum roots by Ramaiah et al (1991), found that 63% of 
the Striga seeds germinated within 24 hours of inoculation of sorghum 
roots.  However a small percentage actually establish a parasitic 
connection with the host.  Timing of haustoria formation is critical in 
the establishment of parasite to host connection.  

Evolutionary Significance of Germination Stimulators

   The purpose of these excreted compounds from the hosts perspective is 
still a question  That is, if one is to believe all plant secondary 
compounds have a biological purpose.  Some research has indicated that 
these compounds are allelochemicals.  Sorgoleone has shown some 
herbicidal activity and may function in allelopathy.  However no work 
has been done to investigate the effects of strigol and it's analogs for 
possible plant defense mechanisms.  The use of host defense chemicals as 
feeding cues is not a new evolutionary idea because of its frequent 
occurrence in insect herbivory.  Atsatt (1977), proposed similar 
patterns would be found in parasitic plant relationships.  Parasitic 
plants host relationships was likened to three evolutionary patterns of 
insects response to host plants: resistance to host chemistry, use of 
host defense chemicals as feeding cues, and secondary utilization of 
host defense compounds.  Though not as much work has been undertaken in 
these areas, there is growing evidence for this hypothesis.

References

Atsatt, P. R. 1977. The insect herbivore as a predictive model in 
	parasitic seed plant biology. Amer. Natur. 111:579-612.

Bebawi, F. F. 1986. Efficacy of ethylene as a germination stimulant of 
	Striga hermonthica seed. Weed Sci. 34:694-698.

Calder, M. and P. Berhardt. (eds.) 1983. The Biology of Mistletoes. 
	Academic Press, Sydney.

Chang, M., and D. G. Lynn. 1986. The haustoria and the chemistry of 
	host recognition in parasitic angiosperms. J. Chem. Ecol. 12:561-
	579.

Chang, M., D. H Netzly., L. G. Butler, and D. G. Lynn. 1986. Chemical 
	regulation of distance: Characterization of the first natural host 
	germination stimulant for Striga asiatica. J. Am. Chem. Soc. 
	108:7858-7860.

Cook, C. E., L. P. Whichard, M. E. Wall, G. H. Egley, P. Coggon, P. A. 
	Luhan, and A. T. McPhail. 1972. Germination stimulants. II. The 
	structure of strigol- A potent seed germination stimulant for 
	witchweed (Striga lutea Lour.). J. Am. Chem. Soc. 94:6198-6199.

Eplee, R. E. 1992. Witchweed (Striga asiatica.): an overview of 
	management strategies in the USA. Crop Protection. 1:3-7.

Gabbar, A., T. Babiker, G. Ejeta, L. G. Butler, and W. Woodson. 1993. 
	Ethylene biosynthesis and strigol-induced germination of Striga 
	asiatica. Physiol. Plant. 88:359-365.

Hauck, C., S. MŸller, and H. Schildknecht. 1992. A germination 
	stimulant for parasitic flowering plants from Sorghum bicolor, a 
	genuine host plant. J. Plant Physiol. 139:474-478.

Hess, D. E., G. Ejeta, and L. G. Butler. 1992. Selecting sorghum 
	genotypes expressing a quantitative biosynthetic trait that 
	confeers resistance to Striga. Phytochem. 31:493-497.

Igbinnosa, I. and S. N. C. Okonkwo. 1992. Stimulation of germination 
	of seeds of cowpea witchweed (Striga gesnerioides) by sodium 
	hyperchlorite and some growth regulators. Weed Sci. 40:25-28.

Logan, D. C. and G. R. Stewart. 1991. Role of ethylene in the 
	germination of the hemiparasite Striga hermonthica. Plant 
	Physiol. 97:1435-1438.

Mangnus E. M., and B. Zwanenburg. 1992. Tentative molecular 
	mechanisms for germination stimulation of Striga and 	Orobanche 
      seeds by strigol and its synthetic analogues. J. Am. 	Chem. Soc. 
      40:1066-1070.

Mangnus E. M., L. A. van Vliet, D. A. L. Vandenput, and B. 	Zwanenburg. 
      1992. Structural modifications of strigol analogues. Influence of 
      the B and C rings on the bioactivity of the germination stimulant 
       GR24. J. Am. Chem. Soc. 40:1222-1229.

MŸller, S., C. Hauck and H. Schildknecht. 1992. Germination 
	stimulants produced by Vigna unguiculata Walp cv Saunders 
	Upright. J. Plant Growth Regul. 11:77-84.

Musselman, L. J. 1980. The biology of Striga, Orobanche, and other 
	root-parasitic weeds. Ann. Rev. Phytopathol. 18:463-489.

Musselman, L. J. (ed.) 1987. Parasitic Weeds in Agriculture. Vol 1. 
	Striga. CRC Press. Boca Raton.

Netzley, D. H., J. L. Riopel, G. Ejeta, and L. G. Butler. 1988. 
Germination 	stimulants of witchweed (Striga asiatica) from 
hydrophobic 	root exudate of sorghum (Sorghum bicolor). Weed Sci. 
36:441-	446.

Olivier, A., N. Benhamou, and G. D. Leroux. 1990. Cell surface 
	interactions between sorghum roots and the parasitic weed 	Striga 
hermonthica : cytochemical aspects of cellulose 	distribution in 
resistant and susceptible host tissues. Can. J. Bot. 	69:1679-1690.

Parker, C. 1991. Protection of crops against parasitic weeds. Crop Pro. 
	10:6-22.

Ramaiah, K. V., V. L. Chidley, and L. R. House. 1991. A time-course 
	study of early establishment stages of parasitic angiosperm 
	Striga asiatica on susceptible sorghum roots. Anna. Appl. Biol. 
	118:403-410.

Siame, B. A., Y. Weerasuriya, K. Wood, G. Ejeta, and L. G. Butler. 1993. 
	Isolation of strigol, a germination stimulant for Striga asiatica, 
  	from host plants. J. Agric. Food Chem. 41:1486-1491.

Steffens, J. C., D. G. Lynn, and J. L. Riopel. 1986. An haustoria 
      inducer for the root parasite Agalinis purpurea. Phytochem.
      10:2291-2298.

Stewart, G. R. and M. C. Press. 1990. The physiology and biochemistry 
	of parasitic angiosperms. Annu. Rev. Plant Physiol. Plant Mol. 
	Biol. 41:127-151.

Vail, S. L., O.D. Dailey, E. J. Blanchard, A. B. Pepperman, and J. L. 
	Riopol. 1990. Terpenoid precursers of strigol as seed 
	germination stimulants of broomrape (Orobanche ramosa) and 
	witchweed (Striga asiatica). J. Plant Growth Regul. 9:77-83.

Weerasuriya. Y., B. Siame, D. Hess, G. Ejeta, and L. G. Butler. 1993. 
	Influence of conditions and genotype on the amount of Striga 
	germination stimulants exuded by roots of several host crops. J. 
	Agric. Food Chem. 41:1492-1496.

Worshum, A. D. 1987. Germination of witchweed seeds.-In Parasitic 
	Weeds in Agriculture (L. J. Musselman. ed.), Vol. 1, Striga. pp. 
	45-61. CRC Press, Boca Raton, FL.