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Intra-ovarian trichomes, mucilage secretion and hollow styles in

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Rudall, PJ., Prychid, C.J. and Jones, C. (1998). Intra-ovarian trichomes, mucilage secretion and hollow
styles in monocotyledons. In: S.J. Owens and PJ. Rudall (Editors). Reproductive Biology, pp.219-230.
Royal Botanic Gardens, Kew.

INTRA.OVARIAN TRICHOMES, MUCILAGE SECRETION

AND HOLLOW STYLES IN MONOCOTYLEDONS

PeuIa J. RUDALL. CHRTSTTNA J. PRYCHID AND CLAIRE JONES

Jodrell Laboratory, Royal Botanic Gardens, Kew, Richmond, Surrey, TW9 3DS, UK

Abstract
Intra-ovarian trichomes are taxonomically widespread in monocotyledons, but especially
clustered among the 'basal' groups, such as Acorils and Araceae. By means of extensive Golgi
activity, they actively secrete a jelly-like mucilage which fills the ovary locules. Since
mucilage is known to promote pollen tube growth, intra-ovarian trichomes have a role in the
reproductive biology of the plants concerned, as specialised transmitting tissues or obturators.
Mucilage-filled ovaries are correlated to some extent with aquatic habitats and orthotropous
ovules. This study also indicates a conelation with short styles, which may be either hollow
(as in most monocotyledons) or solid (as in most dicotyledons). Secretory intra-ovarian
trichomes are rare in dicotyledons, but mucilage-filled ovaries occur in some 'primitive'
magnoliid dicot families closely related to monocotyledons.

Introduction

This paper presents new data on intra-ovarian trichomes (IOTs) and mucilage-filled ovaries in
monocotyledons, and reviews their occunence in a systematic context, in order to test their
homologies and consider their wider implications in terms of breeding systems. IOTs are hairs
that occur within the ovary locules. In monocotyledons they are invariably associated with a
jelly-like mucilage which fllls the entire ovary cavity (Table 1). It has not hitherto been
demonstrated whether the trichomes themselves secrete the mucilage, although this is
generally assumed.
There is strong consensus (e.g. Sassen, 7914) that mucilage has a roie as a nutrient medium
in guiding pollen tubes in both solid and hollow styles, analogous to that of fungal hyphal
growth in vitro. Ovarian mucilage probably has a similar role to stylar mucilage, presumably
with a chemical gradient. French (1987) speculated that inAraceae mucilage bridges the gap
for pollen tube growth between the stylar canal and the micropyle. In the f'ew species where
IOTs are absent, mucilage is also lacking, and in most of these cases there is a single ovule
occupying most of the locule. Similarly, Apparao & Shah (1989) speculated that in Ottelia
alismoides (Hydrocharitaceae) the intra-ovarian mucilage is a medium for free movement of
pollen tubes and their nourishment in the ovarian cavity. Labarca & Loewus (1973)
demonstrated that stylar canal exudates are a major source of carbohydrates for pollen tube
wall biosynthesis rn Lilium. Sanders & Lord (1989) introduced latex beads into the stylar
transmitting tissue of Hemerocallis, Raphanus and Vicia and demonstrated that they were
translocated to the ovules with the same speed as pollen tubes, indicating that pollen tubes are
actively guided by the secretory matrix. In plants with solid styles, including most

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Reproductive Biology

dicotyledons, pollen tubes grow intercellularly between the cells of a central transmitting
tissue, or
sometimes through the cell walls. In plants with hollow styles (i.e. most
monocotyledons), pollen tubes grow along the surface of the secretory stylar canal cells (e.g.
in Gladiolus and Lilium: Clarke er al., 1917), sometimes beneath the cuticle (e.g. in
Hemerocallis: Sanders & Lord, 1992). In the ovary of the date palm (Phoenir), pollen tubes
follow a groove of mucilage to the micropyle (Knox, 1984). Mucilage-secreting IOTs
therefore have a role in the reproductive biology of the plants concerned, as specialised
transmitting tissues or obturators.

Material and Methods

Perhaps the main reason for the paucity of information on IOTs is the notorious difficulty
in obtaining good sections of mucilage-filled flowers (e.g. of Araceae: Eyde et al. 1967).
The carbohydrate component of the mucilage causes the ovaries to harden during
dehydration. Aldehydes used in conventional fixation stabilise proteins by forming cross
links with them, particularly with the e-NH2 groups of lysine side chains. Some lipids also
appear to be included in the fixation. Osmium tetroxide used as a secondary fixative in
electron microscopy work stabilises the majority of lipids and it is hoped that between them
some carbohydrates will be fixed. However, if the mucilage is predominantly carbohydrate,
as in Astelia grandis and Collospermum hastatum (this paper), there is nothing to preserve
it and it will either flow out of the ovary and be lost or will remain there and act as a barrier
to the penetration of the fixative into the ovarian cavity, resulting in degradation of the
ovules and IOTs.

Material examined
Material examined was coliected from the living collections at the Royat Botanic Gardens,
Kew (HK + garden accession number) or from wild-source material.
Acoraceae: Acorus calamus L. 'Variegatus' (HK 1969-19633), A. gramineus Soland.
'Variegatus' (HK 1969-18000), A. gramineus f . minor Hort. (HK 1969-17999).
Asteliaceae: Astelia grandi.i Hook. f. ex T. Kirk (HK 1972 3488): Collospermum hastatum
(Collenso) Skottsb. (HK 1986-3592).
Araceae: Aglaonema modestum Schott. ex Engl. (HK 1965-22401), A. stenophyllurz Merill
(HK 1982-5008); Alocasia micholitziana Sander (HK 1980-1232); Anthurium andraeanum
Linden (HK 1960-31808); Arisarum vulgare Targ. Toz. (HK 1912-10427); Arum dioscoridis
Sibth.& Sm. var. dioscoridis (HK 1979-2190), A. rupicola Boiss. var. rupicola (HK
1985-2128); Stenospermation uleiK. Krause (HK 1990 2738).
Hanguanaceae'. Hanguana malayana Merrill (s.n.).

Transmission Electron Microscopy (TEM)

Gynoecia were cut from the flowers and placed immediately into two different flxatives:
Karnovsky's fixative (2Va paraformaldehyde and 2.5Vo glutaraldehyde) (Glauert, i975) and
acrolein (27o)-glutaraldehyde (2.5Vo) in 0.1M phosphate buffer, pH 7.2. Tissues were dissected
whilst under fix, leaving 2mmr pieces, and placed under vacuum for 30 minutes. Primary
fixation was then continued for 2.5 hours at room temperature and pressure. After washing in
buffer the samples were post-fixed rn 17o osmium tetroxide for two hours, rinsed, dehydrated
in an ethanol series and embedded in LR White resin (medium grade). Silver sections were
collected on Formvar-coated slot grids and stained in aqueous uranyl acetate and lead citrate
(Reynolds, 1963). Photographs were taken on a JEOL JEM i2-10.

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 Intra-ovarian trichomes in monocotyledons

Light Microscopy (LM)

For wax embedding and sectioning, flowers were fixed in Formalin acetic alcohol (FAA)
and stored in'70Vo ethanol. Complete flowers or ovaries were embedded in Paraplast using
standard methods of wax embedding, and serially sectioned using a rotary microtome.
Sections were stained in safianin and Alcian blue, dehydrated through an alcohol series to
100% ethanol then Histoclear, mounted in Euparal, and examined using normal bright-field
optics on a Leitz Diaplan photomicroscope.
For histochemistry the following stains were applied. PAS, periodic acid-Schiff's reaction,
was used as a general polysaccharide stain (Hotchkiss, 1948; McManus, 1948). As a control,
the periodic acid was omitted. The ninhydrin-Schiff's reaction (Yasuma & Ichikawa, 1953)
was used to localise total protein. For controls blocking procedures were applied pdor to the
ninhydrin step. Starch was localised with the IKI reaction (Johansen, 1940).
Semi-thin sections (c. 0.5pm) taken from material embedded for TEM were collected on
slides and stained with the metachromatic stain toluidine blue (0.57o in 0.lM phosphate buffer,
pH 7.0) (Feder & O'Brien, 1968).
Intra-ovarian trichomes are taxonomically widespread in monocotyledons (Table 1),
occurring among several unrelated groups from each of the major clades (sensa Chase et al.
1995), indicating that they have evolved more than once. They occur in the commelinoid
cl.ade (Hanguana and some Zingiberales, such as Musaceae), the asparagoid clade
(Asteliaceae) and the dioscoreoid clade (Pandanaceae). However, they are especially
clustered among the 'primitive' monocotyledons, such as Acorus and Araceae. In contrast to
dicotyledons, monocot IOTs are always associated with mucilage secretion. Mucilage-filled
ovaries are rare in monocotyledons lacking IOTs (Table 1), apart from some 'basal' taxa
(sensa Chase et a1..,1995), such as Ottelia (Hydtocharitaceae), where mucilage is secreted
from intracarpellary cells present in clusters in the ovary wall above the micropyle of each
ovule (Apparao & Shah, 1989).

Structure and taxonomic distribution of IOTs in monocotyledons

TABLE 1. Records of intra-ovarian trichomes and mucilage-filled ovaries in monocotyledons

Family Genera IOTs Mucilage- Reference

fil1ed ovaries

Acoraceae Acorus + + French 1987; this paper

Araceae most genera + + Dalmer 1880; Eyde et al. 1967;
French 1987
Asteliaceae Astelia, Skottsberg 1934; this paper
Collospermum
Hanguanaceae Hanguana + + this paper
Hyacinthaceae Ornithogalum ? + Tilton & Horner 1980 (& see Dalmer I 880)
Hydrocharitaceae Hydrochrtris, + Troll 1931; Endress 1995
Ottelict (+) Apparao & Shah 1989
Lemnaceae Lemna ? Eyde et al. lqoTl Buzgo 199.+:
Endress 1995
Musaceae several genera + e.g. Kirchoff 1992; Alquini 1992
Pandanaceae genera of + Fagerlind 1940; Endress 1995, Cheah &
Pandananae Stone 1975

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Reproductive Biology

In dicotyledons, non-secretory IOTs occur in several families, such as Ericaceae,

Rubiaceae, Saiicaceae, Styracaceae and Sapotaceae (Dickison, 1993). There is relatively
little evidence of secretion from IOTs in dicotyledons, except in rare cases, such as ovarian
glandular trichomes of Acacia (Kenrick & Knox, 1989), and the placental and
integumentary trichomes of some Urticaceae and Euphorbiaceae, which sometimes function
as obturators (Maheshwari, 1950). However, mucilage-filled ovaries occur in some
Magnoliid dicotyledon families that are considered closely related to monocotyledons, such
as Nymphaeaceae, Cabombaceae (Endress, 1990, 1994) and Chloranthaceae (Endress,
1987), where mucilage is secreted directly from the ovary wall rather than from IOTs. IOTs
therefore probably evolved from secretory gynoecial epidermal cells (probably several times
in monocotyledons), the presence of mucilage-filled ovaries being probably the ancestral
(plesiomorphic) state for this character.
According to Endress (1990, 1995) mucilage-filled ovaries occur mainly in aquatic plants or
plants of moist habitats, indicating that a constant water supply is necessary for this structure.
He also suggested a correlation between orthotropous ovules and mucilage-mediated pollen
tube transmission, as there is less constraint on ovules of these species to be curved and to
direct the micropyle towards the placenta. He gave examples from the dicot family
Nymphaeaceae (Barclaya) and the monocot Hydrocharitaceae (Hydrocharls). This conelation
certainly holds true for many Araceae, and some other monocot taxa, such as Hanguana,
which is common in swampy areas, and has orthotropous ovules (Fig. 1B). Astelia and
Collospermum are not specifically swamp-dwelling, and indeed inctude several epiphytes, but
may prefer wetter habitats.
We have also observed some congruence between intra-ovarian trichomes and shoft styles or
almost sessile stigmas (lacking styles), especially in Acoraceae (Fig. 1D), Asteliaceae (Fig. 1A),
Araceae, Lemnaceae and Hanguana (Fig. 1B), although some other taxa with short styles lack
IOTs or mucilage-filled ovaries (e.9. Ra.icas and Xanthorrhoea). Perhaps a combination of the
two factors (wet habitats and shotl styles) may encourage the development of mucilage-secreting
IOTs, in a group already disposed to the production of mucilage for pollen tube growth.
Structure and distribution of IOTs is very similar in all monocotyledons examined. They
are usually unicellular, aithough French (1987) reported some bicellular or multicellular hairs
in a few Araceae. They emerge from the placental region above and below the ovules and
sometimes from base of each funicle, and extend around the ovules, overarching the
micropyles (Fig. 1). Other epidermal cells of the ovary surrounding the loculi are generally
non-secretory, although in some taxa we have noted that they are secretory around the base
of the sty1e. In some taxa (e.g. Astelia: Fig. 1A) IOTs also extend upwards into the style,
effectively forming a 'solid' style, as in most eudicots. This is in contrast to most other
monocotyledons, where styles are hollow, although there are occasional exceptions to this;
for example solid styles occur in some Poaceae and all Atliaceae sensu stricto (Fay and
Chase, 1996).

Evidence for secretory activity

Although Engler (1920) considered Ihal in Anthurium and Lagenandra mucilage formation
results from hair breakdown, our results demonstrate that, at least in Acorus and Araceae, IOTs
actively secrete the mucilage, through extensive Gotgi (dictyosome) activity.
In flowers of Acorus and Araceae just prior to anthesis (Figs. 2-4), when mucilage is
copious, the IOTs have dense cytoplasm, prominent nuclei and small vacuoles often
containing a granular or fibrous material (mucilage). Lipid bodies are also present. Large
amounts of endoplasmic reticulum, often with a swollen appearance, are present throughout
the cytoplasm (Fig. 3C), together with several mitochondria and a number of dense
dictyosomes with swollen cisternae on their maturing faces. Along part of their perimeter, the

222
 Intra-ovarian trichomes in monocotvledons

dictyosome cisternae are associated with a population of small vesicles which are coated on
their outer surfaces (Figs. 3D, 4A-C). The cisternae bud off from the Golgi stack to form
vesicles which then migrate through the cytoplasm and fuse with the plasma membrane to
release their secretory contents (Fig. aD). Mucilage accumulates between the protoplast and
the cell wall (Fig. 3A), which is relatively thick, before diffusing into the ovarian cavity
(exocytosis). Accumulated mucilage is lacking from other ovary or ovule cel1s lining the
ovary cavity (which are relatively thin-wa11ed), indicating that the IOTs are the primary
source of the massive mucilage deposition in the locule.
The presence of large numbers of Golgi bodies associated with small vesicles is consistent
with secretory activity, e.g. in legume stigmas (Kenrick & Knox, 1989), stylar canal cells of
Trimez.ia (Bystedt & Vennigerholz, 199I) and Ornithogalum (TlIton & Horner, 1980) and
transmitting tissue of Primula (Heslop-Harrison et al, 1981). In mucilage cells of Opuntia
(Fahn 1979), mucilage is produced in Golgi bodies in the cytoplasm and accumulates between
the protoplast and the cell wall.
Rough endoplasmic reticulum (Fig. 3C) is also thought to play a part in the synthesis of
mucilage (Bouchet & Deysson, 1974; Horner & Lersten, 1968) and nectar. Swollen
cisternae bud off to form vesicles which then fuse with the plasmalemma and eliminate their
substances from the cytoplasm. Multivesicular bodies thought to originate liom
endoplasmic reticulum (Jensen, 1965; Bystedt & Vennigerholz, 1991) and dicryosomes
(Sedgley & Clarke, 1986; Miki-Hirosige et al, 1987) are involved in the transportation of
secretion in various tissues, such as transmitting tissue of Lycopersicon (Cresti et aL,1976)
and papillar cells of Hordeum and Secale (J. Heslop-Harrison & Y. Heslop-Harrison, 1980).
It has also been postulated that mitochondria are involved in mucilage production (Werker
& Kislev, 1978).
Ovary cells at the base of the trichomes contain numerous starch grains (Fig. 28) which
presumably act as a nutrition reserve and become depleted during mucilage production, as
Apparao & Shah (1989) also noted in intra-ovarian mucilage-secreting cetls of Ottelia
alismoides. Starch grains are absent from the trichomes themselves. Loss of starch grains
from plastids is thought to result from breakdown into monomeric or dimeric sugars which
then leave the cells by eccrine secretion (Kandasamy & Kristen, 1987; Bystedt &
Vennigerholz, 1 99 1 ).
In older IOTs towards the end of their secretory life there are large vacuoles often containing
fibrous mucilage. These are surrounded by ribosome-rich cytoplasm containing a single large
nucleus, endoplasmic reticulum and dictyosomes, but by this stage there are often relatively
few mitochondria. Sometimes hypertophied Golgi bodies occupy a great part of the cytoplasm,
togther with occasional spherical bodies (Fig. 2.A) that may represent autophagic vesicles,
which are commonly fbund in autolysing secretory cetls (e.g. Nair el al., 1983).

Nature of the secretion product

The mucilage itself is often fibrous, indicating that outer wall fibrils are being sloughed off
together with the mucilage. This often results in poor definition of the boundary between the
exudate and the secretory walls, the wall and the adjacent layer of secretion having a similar
structure (see also Bell & Hicks, 1976; Hill & Lord, 1987; Sanders et al, 1990; Sanders &
Lord,1992). At high concentrations mucilage fibrils aggregate laterally to form a web-iike
network. At lower concentrations globular particles in the secretion aggregate in a linear
fashion. Bystedt & Vennigerholz (1991) also reported a separation of the wall fibrils in
secretory gynoecial ce1ls of Trimezia, as secretion passed through them, and in stylar canal
cells of Lilium the wall layer becomes a component of the secretion (Hu er al., 1982;. The
developing testa epidermis of Linum seeds produces a fibrillar, predominantly carbohydrate
mucilage (Heinz, 1984).

223
Reproductive Biology

c
FIG 1 . Light micrographs (LM). A. A.stelia hanksii, LS gynoecium; ovary lovules fllled with mucilage

(m). B. Hanguunu malayantt, LS f'emale flower with short, broad gynoecium and nectariferous
stanrinodes. C. Astelia bcutksii, TS trilocu lar ovary with rnucilage Gn) filling ovary locules. D. Acorus
gramineus f. ntinor, LS gynoecium. iot = intra-ovarian trichomes. m = mucilage. ov = ovule. st =
stigma. Scale bars = 100mm.

224
 Intra-ovarian trichomes in monocotvledons

ffi
skjf
t\x
t!.,r
w.,
\lirs

Ftc.2. Transmission electron micrographs (TEM) (composite picture in C). A. Stenosperm.ation ulei,TS
through IOTs. 8.. C. Akx:asia ni.ch.olitiana. B. Ovarian cells at base ol'IOTs containing large starch
plastids (s). C. IOTs with denser cytoplasm than basal ovary tissue. Fibrous mr,rcilage (r.r.r) in ovary
locule. Scale bars = 5mm.

225
Reproductive Biology

,$r,

FIG 3. Transmissiorr electron rnicrographs (TEM). A. Slenospernultidr a1al, fibrous mucilage (n'r) present
in periplasmic space. B.-D. Acortts gramineus.l. ntinor B. IOT with cytoplasm dominated by
secretory vesicles ancl endoplasmic reticulurn. C. Several profiles of endoplasmic reticulum (tmowed)
and theil associated ribosomes. D. Dictyoson.re with individual cisternae marginally vesiculated.
Sorne ofthe swollon cisternae may have already sloughed ofTto fbrm secretory vesicles. Scale bars:
in A.. C.. D. = lmm. B. = 5mm.

226
 Intra-ovalian trichomes in monocotyledons

FtG.4. Acorus gruminetLs J: nt.inor (TEM). A.. B. Cytoplasm dominated by dictyosomes (d) and secretory
vesicles (v). C. Secretory vesicles and space between plasma membrane and cell wall containing a
substance (arrowed), presurnably rlucilage. D. Secretory vesicle fused with plasma membrane,
releasing its contcnts into the penplasmic space. Scale bars = I mm.
Reproductive Biology

Histochemical studies on ovarian mucilage of our matertal of Astelia grandis and

Collospermum hctstatwn indicated the presence of insoluble polysaccharides and
mucopolysaccharides (carbohydrates) and the absence of proteins and iipids. This coincides
with observations by Apparao & Shah (1989) on ovarian mucilage in Ottelia
(Hydrocharitaceae) and Bystedt & Vennigerholz (1991) on gynoecial secretions of Trimez.ia
.fbsteriana (Iridacezre). Sanders & Lord (1992) and Knox (1984) reported a variety of
component compounds of gynoecial secretion products (extracellular matrices: ECMs),
including sugars, amino acids, peptides, phenolics, fatty acids, lipids and high molecular weight
polysaccharides involving mucilages, proteoglycans and pectic compounds and complex
proteins, such as glycoproteins and lipoproteins, the exact composition varying between
species. In many plants secretion products are chemically different between stigma and stylar
canal cells, indicating a difference in function (Labarca & Loewus, 1973; Kenrick & Knox,
1989). Whereas stylar and ovarian secretions guide the pollen tubes (see below), stigmatic
secretions may have a selective role in self-compatible crosses (Sanders & Lord, 1992).
The secretion product from stylar canal cells is often described as mucilage. The
transmitting tissue of the solid styles of dicotyledons has an mucilagenous intercellular
matrix which Sassen (1974) and Cresti et al. (7916) identified as a complex substance
containing pectin. This is comparable to the mucilage found in the canal of hollow styles
(Labarca et al.,1970).

References

Alquini, Y. (1992). Anatornia dos drgaos em desenvolvomento de Musa rosacea Jacq.

(Musaceae). PhD Thesis, unpublished, University of Sao Paulo.
Apparao, B.J. and Shah, C.K. (1989). Intracarpellary secretory cells and theirrole in Ottelia
alismoides. .1. Indion Bot. Soc. 68:25-28.
Bell, J. and Hicks, G. (1916). Transmitting tissue in the pistil of tobacco: light and electron
microscopic observations. Planta l3l : I 87-200.
Bouchet, P and Deysson, G. (1974). Les canaux )r mucilage des angiospermes. Etude
morphologique et ultrastructurale des cellules constituant les canaux d mucilage du
Sterculia bidwilli Hook. Rev. Gdn. tsot. 8l 369402.
Bystedt, P.-A. and Vennigerholz, F. (1990). The transmitting tract tn Trimezia fosteriana
(Iridaceae). I. Ultrastructure in the stigma, style and ovary. Nordic J. Bot.9:507-518.
Chase, M.W., Stevenson, D.W., Wilkin, P. and Rudall, P. (1995). Monocot systematics: a
combined analysis. In: PJ. Rudall, P.J. Cribb, D.F. Cutler and C.J. Humphries (editors).
Monocotyledons: systematics and evolution, pp. 685-730, Royal Botanic Gardens, Kew.
Cheah, C.H. and Stone, B.C. (1975). Embryo sac and microsporangium development in
Pandunt s (Pandanaceae) . P hy t o mo rp ho I o gy 25 : 228-238.
Clarke, A.E., Considine, J.A., Ward, R. and Knox, R.B. (1977). Mechanism of pollination in
Gladiolus: roles of the stigma and pollen-tube guide. Ann. Bot. 4l: 75-20.
Cresti, M., van Went, J.L., Pacini, E. and Willemse, M.T.M. (1976). Ultrastructure of
transmitting tissue of Lycopersicon peruvianum style: development and histochemistry.
Planta 132: 305-3 12.
Dalmer, M. (i880). Ueber die Leitung der Pollenschlauche bei den Angiospermen. Jenaische
Z. Natw'wiss. 14: 530-566.
Dickison, W.C. (1993). Floral anatomy of the Styracaceae, including observations on intra-
ovarian trichomes. Bot. J. Linn. Soc.7l2:223-255.
Endress, P.K. (1987). The Chloranthaceae: reproductive structures and phylogenetic position.
Bot. Jahrb. Sysr. 109: 153-226.

228
 Intra-ovarian trichomes in monocotvledons

Endress, P.K. (1990). Evolution of reproductive structures and functions in primitive

angiosperms (Magnoliidae). Mem. New York Bot. Gard. 55:- 5-34.
Endress, P.K. ( I 994). Floral structure and evolution of primitive angiosperms: recent advances.
P/. Sys/. Evol. 192:.79-97.
Endress, P.K. (1995). Majorevolutionary traits of monocotflowers. In: P.J. Rudall, p.J. Cribb,
D.F. Cutler and C.J. Humphries (editors) Monocotyledons: systematics and evolution, pp.
43 79, Royal Botanic Gardens, Kew.
Eyde, R.H., Nicolson, D.H. and Sherwin, P. (1967). A survey of floral anatomy in Araceae.
Amer. J. Bot. 54: 478491 .

Fagerlind, F. (1940). Stempelbau und Embryosackentwicklung bei einigen Pandanazeen. Ann.

Jard. Bot. Btdtenz.org 49 55-18.
Fahn, A. (1979). Secretory tissues in plants. Academic Press, London.
Fay, M.F. and Chase, M.W. (1996). Resunection of Themidaceae for the Brodiaea alliance. and
recircumscription of Alliaceae, Amaryliidaceae and Agapanthoideae. Taxon 45: 441451.
Feder, N. and O'Brien, T.P. (-l968). Plant microtechnique: some principles and new methods.
Amer J. Bor. 55: 123-142.
French, J.C. (1987). Structure of ovular and placental trichomes of Araceae. Bot. Gaz. 148:
1 98-208.

Glauert, A.M. (1975). Fixation, dehydration and embedding of biological specimens. Norlh-
Ho1land, Amsterdam. (Practical methods in electron microscopy, vol. 3, pt. 1)
Heinze, U. and Amelunxen, F. (1984). Ztr Schleimbildung in Linum-Samen.
Elektronenmikroskopische und chemische Analysen. Ber. Deutsch. Bot. Ges.97 451464.
Heslop-Harrison, J. and Heslop-Harrison, Y. (1980). The pollen,stigma interaction in the
grasses. I. Fine-structure and cytochemistry of the stigmas of Hordeum. and Secale. Acta
Bot. Neerl. 29: 261-27 6.
Heslop-Harison, Y., Heslop-Harrison, J. and Shivanna, K.R. (1981). Heterostyly tn primula.
I. Fine-structural and cytochemical features of the stigma and style in Pritnula vulgaris
Hlcls. Protoplasma 107: I1 l-187 .
Hill, J.P. and Lord, E.M. ( 1987). Dynamics of pollen tube growth in the wild radtsh Raphanus
raphanistrum (Brassicaceae). II. Moryhology, cytochemistry and ultrastructure of
transmitting tissues, and path of pollen tube growth. Amer. J. Bot. 74: 988-99i.
Horner, H.T. and Lersten, N.R. (1968). Development, structure and function of secretory
trichomes in Psychotria bacteriophila (Rubiaceae). Amer. J. Bot. 55:1089-1099.
Hotchkiss, R.D. (191.8). A microchemical reaction resulting in the staining of polysaccharide
sftuctures in fixed tissue preparations. Arch. Biochem. 16: 131-141.
Hu, S., Zhu, C. and Xu, L. (1982). The structure of the canal cell in the slyle of Lilium regale.
Acta Bot. Sin. 24:395-406.
Jensen, W.A. (1965). The composition and ultrastructure of the nucellus in cotton.
J. Ubrastruct. Res.13: l12-128.
Johansen, D.A. (1940). Plant microtechnique. McGraw-Hilt, New York.
Kandasamy, M.K. and Kristen, U. (1987). Developmental aspects of ultrastructure,
histochemistry and receptivity of the stigma of Nicotiana sylvestris. Ann. Bot. 60 427 437 .
Kenrick, J. and Knox, R.B. (1989). Pollen-pistil interactions in Leguminosae (Mimosoideae).
In: C.H. Stirton and J.L.Zartcchi (editors). Advances in legume biology, pp. 127-156,
Missouri Botanical Garden, St. Louis, Mo. (Monographs in systematic botany from the
Missouri Botanical Garden, 29)
Kirchoff, B.K. (1992). Ovary sttucture and anatomy in the Heliconiaceae and Musaceae
(Zingiberales). Canad. J. Bot. 70 2490-2508.
Knox, R.B. (1984). Pollen-pistil interactions. In: H.F. Linskens and J. Hestop-Harrison
(editors). Cellular interactions, pp. 508-608, Springer Verlag, Berlin. (Encyclopedia of
plant physiology, vol. 17).

229
Rcproductive Biology

Labarca, C. and Loewus, F. (1973). The nutritional role of pistil exudate in pollen tube wall
formation in Lil.ium longiflorum.II Production and utilization of exudate from stigma and
stylar canal. Pl. Physiol. 52:87-92.
Labarca C., Kroh M. and Loewus, F. (1970). The composition of stigmatic exudate from
Liliunt longifLorum. Labelltng studies with myoinositol, D-glucose, and L-proline. Pl.
Physiol.46:150 l5b.
Maheshwari, P. (1950). An introduction to the embryology of angiosperms. McGraw-Hil1,
New York.
McManus, J.F.A. (1948). Histological and histochemical uses of periodic acid. Stain Technol.
23: 99-108.
Miki-Hirosige, H., Hoek, LH.S. and Nakamura, S. (1987). Secretions from the pistll of Lilium
l.ongifl.ctrum. Amer. J. Bot.74: 1709 1'715.
Nair, M.N.B, Shah, J.J. and Subramanyam, S.V. (1983). Ultrastructure and histochemistry of
traumatic gum ducts in the wood of Azadirachta indica A. Juss. 1AIIA Bull. 4: 103-112.
Reynolds, E.S. (1963). The use of lead citrate at a high pH as an electron opaque stain in
electron microscopy. J. Cell Biol. 17:202-212.
Sanders, L.C. and Lord, E.M. (1989). Directed movement of latex particles in the gynoecia of
three species offlowering plants. Science 243: 1606-1608.
Sanders, L.C. and Lord, E.M. (1992). A dynamic role for the stylar matrix in pollen tube
extension. Int. Rev. Cytol. 140:297-318.
Sanders, L.C., Eckard, K.J. and Lord, E.M. (1990). Divergent pollination systems in the
cleistogamous species Collomia grandiflora (Polemoniaceae). Protoplasma 159 26 34.
Sassen, M.M.A. (1974). The stylar transmitting tissue. Acta Bot. Neerl. 23:99-108.
Sedgiey, M. and Clarke, A.E. (1986). Immuno-gold localisation of arabinogalactan protein in
the developing style of Mcorlana alata. Nordic J. Bot. 6: 591,598.
Skottsberg, C. (1934). Studies in the genus Asrelia Banks et Solander. Kongl. Svenska
Vetenskttps akad. Handl., Ser. 3, 14(2): 106pp.
Tilton, VR. and Horner, H.T. (1980). Stigma, style and obturator of Ornithogalum caudatum
(Liliaceae) and their function in the reproductive process. Amer. J. Bot. 67:1113-i131.
Troll, W. (1931). Beitr:ige zur Morphologie des Gynaeceums. I. Uber das Gynaeceum von
Hydrocharitaceen. Planta 17 : 453460.
Werker, E. and Kislev, M. (1978). Mucilage on the root surface and root hairs of Sorghum:
heterogeneity in structure, manner of production and site of accumulation. Ann. Bot. 42:
809 816.
Yasuma, A. and Ichikawa, T. (1953). Ninhydrin-Schiff and alloxan-Schiff staining. A new
histochemical staining method for proteins. J. Lab. Clin. Med. 4l 296-299.

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