Secreting glandular trichomes: more than just hairs

Secreting glandular plant trichome types which accumulate large quantities of metabolic products in the space between their gland cell walls and cuticle permit the plant to amass secretions in a compartment that is virtually outside the plant body. These structures not only accumulate and store what are often phytotoxic oils but they position these compounds as an apparent first line of defense at the surface of the plant. Recent advances in methods for isolation and study of trichome glands have allowed more precise analysis of gland cell metabolism and enzymology. Isolation of mutants with altered trichome phenotypes provides new systems for probing the genetic basis of trichome development. These advances and their continuation can pave the way for future attempts at modification of trichome secretion. The biochemical capability of glandular secreting trichomes and the potential for its future manipulation to exploit this external storage compartment is the focus of this review.     

Development and Structure of Internal Glands and External Glandular Trichomes in Pogostemon cablin

Pogostemon cablin possesses two morphologically and ontogenetically different types of glandular trichomes, one type of bristle hair on the surfaces of leaves and stems and one type of internal gland inside the leaves and stems. The internal gland originates from elementary meristem and is associated with the biosynthesis of oils present inside the leaves and stems. However, there is little information on mechanism for the oil biosynthesis and secretion inside the leaves and stems. In this study, we identified three kinds of glandular trichome types and two kinds of internal gland in the Pogostemon cablin. The oil secretions from internal glands of stems and leaves contained lipids, flavones and terpenes. Our results indicated that endoplasmic reticulum and plastids and vacuoles are likely involved in the biosynthesis of oils in the internal glands and the synthesized oils are transported from endoplasmic reticulum to the cell wall via connecting endoplasmic reticulum membranes to the plasma membrane. And the comparative analysis of the development, distribution, histochemistry and ultrastructures of the internal and external glands in Pogostemon cablin leads us to propose that the internal gland may be a novel secretory structure which is different from external glands.     

Light, conventional and environmental scanning electron microscopy of the trichomes of Cucurbita pepo subsp. pepo var. styriaca and histochemistry of glandular secretory products

BACKGROUND AND AIMS: In the present study, the differences between glandular and non-glandular trichomes, the secretory process and the method of secretion were studied. Previous studies on leaves of Styrian oil pumpkin (Cucurbia pepo var. styriaca) plants have shown that four morphologically and ontogenetically independent glandular and non-glandular trichome types and one bristle hair type can be distinguished. The four types of trichomes can be categorized into three glandular trichome types: type I, a short-stalked trichome with four head cells including a 'middle-cell', two stalk cells and one basal cell; type II, a long-stalked trichome with two head cells, a 'neck-cell' region and a long stalk area; type IV, a 'stipitate-capitate' trichome with a mesophyll cell basement, a short stalk and a multicellular head; type III, a non-glandular 'columnar-digit' trichome, which consists of two head cells continuous with three-celled stalk, and the basal cell. METHODS: The histochemical studies (the main classes of metabolite in secreted material of glandular trichomes) were conducted in fresh and fixed hand sections, using the following tests: Sudan black B, Nile blue A, osmium tetroxide, neutral red, Naturstoffreagent A, FSA (fuchsin-safranin-astra blue), NADI (naphthol + dimethylparaphenylenediamine) and ruthenium red. Each suggested differences in the intercalations during the ontogenetical development of each trichome during the development stage. KEY RESULTS: The histochemical reactions revealed the main components of the materials secreted by all types of trichomes, which include lipids, flavones and terpenes and the different cell wall compositions. Glandular secretions were observed during environmental scanning electron microscopy (ESEM) and the trichomes compared with those seen by conventional scanning electron microscopy (CSEM). CONCLUSIONS: Scanning electron microscopy and histochemical analysis demonstrated that each of the trichomes studied produced and released secretory products in a characteristic way.     

New approaches for studying and exploiting an old protuberance, the plant trichome

BACKGROUND AND AIMS: Much recent study of plant trichomes has focused on various aspects of glandular secreting trichomes (GSTs) and differentiation of simple trichomes. This Botanical Briefing will highlight: research on various aspects of, and manipulation of glandular secreting trichomes; molecular aspects of the differentiation and development of simple trichomes of arabidopsis and cotton; how methods for manipulation of model systems used in the above work can be applied to expand our understanding of less studied surface structures of plants. SCOPE: The Briefing will cover: established and suggested roles of simple and glandular secreting trichomes; recent results regarding solute and ion movement in trichomes; methods for isolating trichomes; recent studies of trichome differentiation and development; attempts to modify metabolism in secreting trichomes; efforts to exploit trichomes for commercial and agronomic purposes.     

龙葵腺毛发育的细胞学研究

利用光学显微镜、扫描电镜和透射电镜技术,观察了龙葵“四叶一心”期时叶片及茎表皮的腺毛的种类、分布,探究了不同类型腺毛的起源、生长、成熟、分泌、衰老等发育过程的细胞学特征;通过组织化学染色和荧光显微技术,观察了龙葵腺毛成分、分布,为龙葵的进一步开发利用提供参考。结果表明：（1）龙葵腺毛分为单细胞头腺毛和多细胞头腺毛两类,前者主要分布于茎表面和叶上下表皮,后者主要分布于茎表面的单细胞头腺毛之间、叶脉及叶边缘;（2）龙葵腺毛发育起始于表皮细胞突起,单细胞头腺毛行顶端生长,具1-4个柄细胞,四种类型;多细胞头腺毛可再分为一层、两层与三层多细胞头腺毛,另具三种特殊类型;（3）龙葵成熟腺毛具分泌能力,通过皮下空间的物质积累导致腺毛头细胞表面形成突起、包块、破口,最终释放分泌物;而头细胞与柄细胞随即皱缩、衰老。（4）超微结构显示,腺毛头细胞中内质网与高尔基体极为丰富,合成代谢及分泌活动活跃,产生大量包裹嗜锇物质的囊泡,囊泡与细胞壁融合,进而将嗜锇物质转移至细胞壁并积累,随后储存在角质层下的皮下空间直至分泌释放;（5）组织化学染色结果表明,腺毛含有萜类、生物碱、脂类、蛋白质、酚类和多糖。头细胞中主要含有萜类、生物碱、脂类、蛋白质、酚类和中性多糖;柄细胞中主要含有萜类、生物碱、脂类。     

Glandular trichomes as a barrier against atmospheric oxidative stress: Relationships with ozone uptake,leaf damage,and emission of LOX products across a diverse set of species

There is a spectacular variability in trichome types and densities and trichome metabolites across species, but the functional implications of this variability in protecting from atmospheric oxidative stresses remain poorly understood. The aim of this study was to evaluate the possible protective role of glandular and non‐glandular trichomes against ozone stress. We investigated the interspecific variation in types and density of trichomes and how these traits were associated with elevated ozone impacts on visible leaf damage, net assimilation rate, stomatal conductance, chlorophyll fluorescence, and emissions of lipoxygenase pathway products in 24 species with widely varying trichome characteristics and taxonomy. Both peltate and capitate glandular trichomes played a critical role in reducing leaf ozone uptake, but no impact of non‐glandular trichomes was observed. Across species, the visible ozone damage varied 10.1‐fold, reduction in net assimilation rate 3.3‐fold, and release of lipoxygenase compounds 14.4‐fold, and species with lower glandular trichome density were more sensitive to ozone stress and more vulnerable to ozone damage compared to species with high glandular trichome density. These results demonstrate that leaf surface glandular trichomes constitute a major factor in reducing ozone toxicity and function as a chemical barrier that neutralizes the ozone before it enters the leaf.     

GLANDULAR STRUCTURES OF HOLOCARPHA AND THEIR ONTOGENY

Carlquist , Sherwin . (Rancho Santa Ana Botanic Garden, Claremont, California.) Glandular structures of Holocarpha and their ontogeny. Amer. Jour. Bot. 46(4): 300–308. Illus. 1959.—Two types of advanced glandular structures occur in the 4 species of the genus Holocarpha. Sessile disk-shaped glands occur at the tips of upper leaves and of involucral and receptacular bracts. Unlike all other glandular structures of Madinae which have been investigated, these originate from several protodermal initials rather than a single one. These glands, however, represent modifications of a glandular trichome. The other type of glandular structure, termed hollow-stalked trichome here, occurs on the outer surface of involucral and receptacular bracts. These trichomes originate from a single cell but differ from others in the formation of a hollow stalk, the wall of which is one cell in thickness. Mesophyll of the bract, often with an included vascular bundle, is present as an intrusion into the base of the hollow stalk. Corresponding to the advanced nature of the glandular structures, the leaves show specializations in the “inrolling” of margins. Upper leaves have a cylindrical organization of vascular tissue, whereas basal leaves are “normal” and leaves of the main stem are intermediate. The species of Holocarpha differ in certain details of leaf anatomy and structure of hollow-stalked trichomes. The systematic distribution of these is given. The essential unity of the various glandular structures of Madinae is discussed both in terms of mature structure and ontogeny, and the steps in the evolution of these are suggested.     

Novel reports of glands in Neotropical species of Indigofera L. (Leguminosae,Papilionoideae)

By considering controversial discussions in the literature with regard to gland denomination in Indigofera species, as well as the taxonomic value of secretory structures in Leguminosae, we aim to morphologically detail glands that had been previously observed in I. microcarpa and I. sabulicola, and to investigate the occurrence of glands in vegetative and reproductive organs of other six Neotropical species that belong to the genus. Glands analyzed through scanning electronic microscopy (SEM) in combination with anatomic analyses correspond to secretory trichomes that are classified into seven types. Main variations in relation to types occurred with regard to head shape and peduncle size. Trichome heads were multicellular, with a thin cuticle. Hollow heads with conspicuous inner space characterized only one type (type 1); the other trichome types had massive heads. Peduncles, which varied from biseriate to multiseriate, had thick, pecto-cellulosic cell walls. Trichomes were found on stems, stipules, petioles, rachis, petiolules, leaflets, bracteoles, sepals, standards and fruits, more commonly along the margins. Each of the eight Indigofera species analyzed had at least two different trichome types out of the seven types that occurred in reproductive and vegetative organs of these taxa. Various types of secretory trichomes were found in I. campestris, I. lespedezioides, I. microcarpa, I. spicata, I. suffruticosa and I. truxillensis. Stems and rachis were the vegetative organs in which a greater variety of trichomes occurred, and sepals were parts of reproductive organs with the same status. Five out of the seven secretory trichome types occurred on both vegetative and reproductive organs. Distribution and gland types differed between species and these gland distribution patterns can be used as diagnostic characters. Reports of glands in Indigofera campestris, I. hirsuta, I. lepedezioides, I. suffruticosa, I. spicata and I. truxillensis, their recognition as secretory trichomes, and the morphological variety of types found for such trichomes are novel data for Indigofera.     

Structural and Histochemical Investigation of the Glandular Trichomes of Salvia aurea L. Leaves, and Chemical Analysis of the Essential Oil

Anatomical and histological investigations of the secretoryhairs ofSalvia aurea leaves, and identification of the maincomponents of the essential oil were carried out. Two typesof glandular trichome were found: peltate glands, characterizedby a short stalk and a large six to eight-celled head, and capitatetrichomes which were further subdivided into two kinds, thefirst with a short monocellular stalk and two-cellular head(type I), and the second with a multicellular stalk, a neckcell and a small globose unicellular head (type II). Whereaspeltate glands and type I capitate trichomes were always present,type II capitate glands were not found in all leaf samples.The histochemical study suggested an ‘endodermal’role for the stalk cell (peltate and capitate type I) as wellas for the neck cell (capitate type II), preventing the lossof essential oil. Histological reactions also revealed the complexnature of the material secreted by all types ofS. aurea trichome,including polysaccharides, polyphenols and proteins, in additionto the essential oil. Qualitative and quantitative GC-MS analysisof the essential oil revealed camphor to be the main constituent.The findings are discussed in relation to studies of trichomesfrom other members of the Lamiaceae. Salvia aurea L.; glandular trichomes; histochemistry; essential oil     

GLAND DISTRIBUTION AND CANNABINOID CONTENT IN CLONES OF CANNABIS SATIVA L.

The relationship between glandular trichomes and cannabinoid content in Cannabis sativa L. was investigated. Three strains of Cannabis, which are annuals, were selected for either a drug, a non-drug, or a fiber trait and then cloned to provide genetically uniform material for analyses over several years. The distribution of the number and type of glands was determined for several organs of different ages including the bract and its subtending monoleaflet leaf and the compound leaf on pistillate plants. Quantitation of glands on these structures was integrated with gas chromatographic analyses of organ cannabinoid profiles. A negative correlation was found between cannabinoid content and gland number for each of the three clones. Isolated heads of the capitate-stalked glands also were analyzed for cannabinoid content and found to vary in relation to clone and gland age. These studies indicate that cannabinoids may occur in plant cells other than glandular trichomes. The results of these studies emphasize the need for stringent sampling procedures in micromorphological studies on trichome distribution and analytical determinations of cannabinoid content in Cannabis.     

Evolution of trichome types and its systematic significance in the genus Phlomoides (Lamioideae–Lamiaceae)

Despite a number of recent molecular phylogenetic studies on Phlomoides, in terms of trichome morphology the genus is still among the most poorly studied taxa in the family Lamiaceae. In order to test the utility of trichome characters for delimitation of sections, subsection and species of Phlomoides, we examined trichomes of 64 species representing all recognized sections and subsections using stereomicroscopy and scaning electron microscopy. Two basic types of trichomes could be identified: non‐glandular and glandular. Both trichome types can be simple or branched. The glandular trichomes were sessile, short stalked or long stalked. Different kinds of branched trichomes were observed in most species of P. sect. Phlomoides, i.e. symmetrically stellate, stellate with a central long branch, bi‐ or trifurcate. The species of P. sect. Filipendula were mostly covered by simple trichomes. Moreover, variation in trichome characters appears to have particular value, not only for the classification at sectional or subsectional rank, but also for delimitation of species within each section. For example, all studied species of P. subsect. Fulgentes are characterized by various kinds of stellate trichomes, while the trichome variability in P. subsect. Tetragonae was sufficiently high for species discrimination. An ancestral character state reconstruction was performed in order to investigate the evolution of trichome types and it revealed the following evolutionary trends in trichome characters of Phlomoides: 1) branched trichomes are primitive in Phlomoides as compared to simple ones, 2) long simple non‐glandular trichomes are derived as compared to short simple ones and 3) the presence of stalked glandular trichomes is advanced as compared to subsessile or sessile ones.     

Structure and development of bracteal nectary glands in Aphelandra (Acanthaceae)

A survey of bracteal (extrafloral) nectaries in species of Aphelandra (Acanthaceae) reveals substantial diversity. Each bracteal nectary is an aggregate of individual glands that vary in number, size, and structure among species. Glands contain three cell layers: a palisade-like secretory cell layer, a one-to-many-celled intermediate layer with thickened cell walls, and a foot layer. Members of the A. pulcherrima complex have one of two distinct gland types: relatively small glands with a single-celled intermediate layer or larger glands that have a multicellular intermediate layer. Nectaries composed of small glands are patches of many (>50) glands, whereas those composed of large glands are patches of < 10 glands. Four outgroup species have bracteal nectaries of numerous small glands with pluricellular intermediate layers. Glands of all three types are initiated as single enlarged protodermal cells, and all undergo similar early periclinal divisions; the large-gland type shows greater subsequent enlargement with many more anticlinal divisions. The bracteal nectar glands are interpreted to be homologous with simpler glandular trichomes, and mark a monophyletic lineage within Aphelandra. Comparisons with outgroup species show that both nectary types in the A. pulcherrima complex have diverged from an ancestral condition of numerous small glands with pluricellular intermediate layers. Use of the ontogenetic criterion to polarize gland type within the A. pulcherrima complex would yield erroneous results because evolution has apparently involved a developmental truncation with loss of cell divisions in the intermediate layer of small glands. Comparable nectar glands in more distant taxa are interpreted as remarkable cases of convergent evolution, perhaps from similar trichome precursors.     

Differences in distribution and performance of two sap-sucking herbivores on glandular and non-glandular Datura wrightii

1. Datura wrightii Regel (Solanaceae) is polymorphic with regard to trichome type. Some plants are densely covered with short, non-glandular trichomes, whereas other plants in the same populations possess glandular trichomes that excrete a sticky exudate. The hypothesis that glandular trichomes enhance resistance to all small insect herbivores is evaluated.
2. Field censuses in four southern Californian D. wrightii plant populations revealed that glandular plants are indeed resistant to whitefly spp. (Homoptera: Aleyrodidae). Whiteflies are almost exclusively found on non-glandular plants. In contrast, Tupiocoris notatus (Distant) (Heteroptera: Miridae), another sap-sucking herbivore of similar body size, is found predominantly on plants with glandular trichomes.
3. Laboratory experiments showed that whiteflies are unable to colonize glandular D. wrightii phenotypes. After the whitefly adults had landed on the leaves of these plants, they were trapped in the exudate and died.
4. Tupiocoris notatus adults, on the other hand, laid significantly more eggs on glandular plants. The presence of the exudate was shown to be the cue that determined their choice of glandular plants.
5. In no-choice experiments, T. notatus nymphs reared on glandular plants had significantly higher survival rates and had shorter developmental periods than those raised on non-glandular plants. This, combined with the higher oviposition rates, resulted in higher T. notatus population growth rates on glandular plants than on non-glandular plants.
6. Glandular trichomes are not therefore a universal protection against small herbivores. Differences in distribution over the two plant types within the natural herbivore guild on D. wrightii may, among other selection pressures, contribute to the maintenance of the observed trichome polymorphism.     

Comparative mutagenesis of plant flavonoids in microbial systems.

The plant flavonoids quercetin (3,5,7,3',4'-pentahydroxyflavone), morin (3,5,7,2',4'-pentahydroxyflavone), kaempferol (3,5,7,4'-tetrahydroxyflavone), chrysin (5,7-dihydroxyflavone), fisetin (3,7,3',4'-tetrahydroxyflavone), myricetin (3,5,7,3',4',5'-hexahydroxyflavone), myricitrin (myricetin-3-rhamnoside), hesperetin (3',5,7-trihydroxy-4'-methoxyflavanone), quercitrin (quercetin-3-L-rhamnoside), rutin (quercetin-3-rhamnosylglucoside or quercetin-3-rutinoside), and hesperidin (hesperetin-7-rutinoside) have been assayed for mutagenicity in the Salmonella/microsomal activation system. Quercetin, morin, kaempferol, fisetin, myricetin, quercitrin and rutin were mutagenic in the histidine reversion system with the frameshift strain TA98. The flavonols quercetin and myricetin are mutagenic without metabolic activation, although more effective when a rat liver microsomal preparation (S-9) is included; all others require metabolic activation. Flavonoids are common constituents of higher plants, with extensive medical uses. In addition to pure compounds, we have examined crude extracts of tobacco (snuff) and extracts from commonly available nutritional supplements containing rutin. Mutagenic activity can be detected and is correlated with the flavonoid content.     

线纹香茶菜叶上腺毛发育的细胞学研究

为进行中药溪黄草基原植物的品种鉴定,采用光镜和电镜对线纹香茶菜(原变种)[Isodon lophanthoides var.lophanthoides]叶上腺毛的发育进行细胞学研究。结果表明,线纹香茶菜具有头状腺毛和盾状腺毛2种类型。头状腺毛无色透明,由1个基细胞、1个柄细胞和1或2个头部分泌细胞构成;盾状腺毛为红色,由1或2个基细胞、1个柄细胞和4~8个分泌细胞构成头部。2种腺毛均由原表皮细胞经两次平周分裂形成,后因柄细胞和头部细胞所处的分化状态不同而形成两类腺毛。2种腺毛超微结构表明,质体、高尔基体和粗面内质网为主要分泌物产生和运输的细胞器。当盾状腺毛成熟时,角质层下间隙充满了分泌物,其分泌物的性质很可能决定了线纹香茶菜腺毛的颜色。     

Glandular trichomes of Ceratotheca triloba (Pedaliaceae): morphology, histochemistry and ultrastructure

This study was initiated to characterize the distribution, morphology, secretion mode, histochemistry and ultrastructure of the glandular trichomes of Ceratotheca triloba using light and electron microscopy. Its leaves bear two morphologically distinct glandular trichomes. The first type has long trichome with 8-12 basal cells of pedestal, 3-14 stalk cells, a neck cell and a head of four cells in one layer. The second type has short trichome comprising one or two basal epidermal cells, a unicellular or bicellular stalk and a multicellular head of two to eight cells. There is a marked circular area in the upper part of each head cell of the long trichome. This area is provided with micropores to exudate directly the secretory product onto the leaf surface by an eccrine pathway. The secretory product has copious amount of dark microbodies arising from plastids which are positive to Sudan tests and osmium tetroxide for unsaturated lipids. The secretion mode of short trichomes is granulocrine and involves two morphologically and histochemically distinct vesicle types: small Golgi-derived vesicles which are positive to Ruthenium Red test for mucilaginous polysaccharides; the second type is dark large microbodies similar to that of long trichomes with low quantity. These two types are stored in numerous peripheral vacuoles and discharge their contents accompanied by the formation of irregular invaginations of the plasmalemma inside the vacuoles via reverse pinocytosis. These two secretion modes of long and short trichomes are reported for the first time in the family Pedaliaceae. The long trichomes have more unsaturated lipids, while the short trichomes contain more mucilaginous polysaccharides.