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Protocol 14 - Herbs and Epiphytes

Maria Aparecida Lopes • E-mail:
Protocol researchers:
East Pará - Dr. João Ubiratan Santos (UFRA)
West Pará - MSc. Chieno Suemtsu (UFOPA)
Amapá - Salustiano da Costa Neto (IEPA)
Mato Grosso - MSc. Ivone Vieira da Silva (UNEMAT/AF)


Estimated species diversity per grid: The families to be considered will be Araceae, Marantaceae and Poaceae: 70 species; aquatic macrophytes and herbaceous phanerogamous aquatic plants from flooded and floodplain areas: 44 species; four classes of pteridophytes, around 15 families, and 15-20 species of pteridophytes, per 5 x 250 m plot.

Biological role of the group: The families of herbs, epiphytes and hemiepiphytes to be studied make up a large proportion of the species contained within the forest’s herbaceous stratum. These are important resources for vertebrate and invertebrate fauna, acting as pollinators and dispersers for a large number of species. Recent studies have utilized herbaceous plants for understanding the structure of tropical communities as well as the geographical distribution standards of biodiversity.

Economical importance of the group: Many species of the above cited families are kept as ornamental plants. In the south of Brazil, this aspect of the species has been widely explored, but in the Amazonian region this potential can be explored even further. Furthermore, some species with resistant fibers can be used in handicrafts; a part of the Amazon region’s economy. The main genera used in handicrafts are: Heteropsis (“Cipó-titica”) from the Araceae family, and Ischnosiphon (“Arumã”) from the Marantaceae family. A number of Araceae genera are used in popular medicine, especially in tropical regions. Crushed leaves from several species of the Anthurium and Montrichardia genera of the previously referred to family, are used as anti-inflammatory medicines and as poultices, respectively. The araceae family is used in the pharmaceutical industry as a source of chemical compounds, such as: saponins, cyanogenic compounds, polyphenols, anthocyanins and flavonoids. There is still a lot to be understood in terms of species distribution, including massive potential for further exploration of the basic ecology, necessary for determining cultivation techniques. Pteridophytes can also be used as indicators of preserved and/or changed environments (colonizing and/or invading). These are normally utilized as ornamental plants and as substrates for the cultivation of orchids and bromeliads (the trunk and tangle of the roots of arborescent ferns). A number of species are used as medicines (anthelmintics), as edible substances (traditionally used in oriental cooking), as well as a number of fern shoots, having a proven carcinogenic action. The term ‘aquatic macrophytes’ was standardized in 1969 by the International Biological Program (IBP) for all types of aquatic plants; from macro-algae to angiosperms that inhabit swamps and other, truly aquatic environments (ESTEVES, 1988). Herbaceous plants that are to be considered within this protocol; types of aquatic macrophytes that live in the water or soils covered with water, or in saturated soils.

Aquatic macrophytes will be ecologically classified in terms of life-form (according to IRGANG et al, 1994), according to their lesser or greater relationship with water, as a result of tidal ranges every 6 hours, defined as follows: a) amphibious – all plants found in saturated and/or flooded soils at each tidal phase; b) emergent – plants that are fixed to permanently submersed substrate, their vegetative and/or reproductive tissues emerging from the water according to tide levels; c) fixed floating – plants that are fixed to the substrate but with their vegetative and/or reproductive tissues floating on the surface; d) free floating – types of plants in which only the roots are immersed in water, with the vegetative and reproductive tissues being emergent; e) fixed submersed - plants that are fixed to the substrate, but always immersed. Depending on the type of water (SIOLI, 1962) and the substrate, aquatic macrophytes are important as pioneering species in the formation of alluvial islands, river banks and streams, as well as being refuges and nurseries for animals within these environments, in addition to helping prevent riverbank erosion. Several aquatic species are important in terms of their potential nutritional, medicinal, fibrous and aromatic uses, for example.

Technique 1. Aquatic macrophytes

The species will be collected within the riparian plots (banks of the aquatic plots used in protocols 6 and 8), at a width which is equal to the length of the area in question, or at a maximum of 10 meters on each side.  Collections will be made in accordance with the life-form in question (Figure 1) according to the gradient of the riverbank, stream, lake, etc…, into the flood zone of the same area; each collected species being registered in terms of life-form, type of water, substrate, geographic coordinates and specific information relative to each plant (Figure 2). In the event that an already collected species is found but in another life-form and/or type of water, it must still be collected and registered. This is in order to keep a record of all of the life-forms for that particular species and, subsequently, the ecological extent of species at any particular level of flooding. Samples must be pressed between sheets of paper at the moment of collection so that no deformations occur because of dehydration. In order to get a good sample, the paper must be changed (facilitating the drying process) before placing the specimens into the drying oven.

Herborization: Species, following conventional methodology, must be collected at a flowering and/or fruiting stage, with relevant information being recorded onto a form (shown in Figure 2). Small specimens should be collected including the root, with the help of pruning shears or other appropriate equipment. At least five samples should be collected, one for each herbarium for which the samples are being collected; one for the group specialist, another specimen for the one of the large national herbaria, with the others being used for the scientific exchange of botanic material occurring between national and foreign herbaria.

In the event of rare material, new occurrences, possible new material, etc, the decision to keep/exchange the sample will be that of the specialist, but not guaranteeing, however, herbaria inclusion.

Sampling unit: a plot of 50 meters.

Field notes: When in the field (and whenever possible) the following information about each plant will be noted down in a standardized field notebook: Collection area, geographic coordinates, collector and collection number, collection data, plant habitat (herb), life-form (amphibian, emergent etc.), height / size of the plant, colour of the flower and fruit, and its usage. The name and number of the collector should be jotted down in pencil on the sheets of paper for all samples of the same type of plant collected in the same location.

Figure 1. Main Biological Life-forms: a) Amphibians; b) Emergents; c) Fixed floating; d) Free floating; e) Fixed submersed; f) Free submersed; g) Epiphyte (IRGANG et al., 1994).

Figure 2. Example of a herbarium record (in Portuguese).

Quantitative procedures

The herbaceous aquatic macrophytes will be sampled using the plot method according to Mueller-Dombois AND Ellenberg (1974).

An area of representative vegetation will be chosen along the banks of the rivers and streams.

A 50 m baseline will be established, accordingly, at which 4 points will be determined, also taking the best representation of the environment into account.

A 20 m area will be measured out at each point, perpendicular to the baseline, or until the edge of the flooded area is reached.

Single m2 plots will be marked out along this 20 m stretch, starting on the left-hand side and continuing every 1 m along the right-hand side, then the left-hand side again, and so on (see Figure 3).

The name of the every species will be noted down in each square, in addition to the land coverage percentage of each species, as well as the percentage of the area that has no vegetation (bare). Coverage levels will be estimated.

Two phytosociological inventories will be made, one during the rainiest period of the year (January to June) and the other during the driest period (July to December). The baseline will be set out in the same location for both of the phytosociological inventories.

Identification of the number of species present and the coverage percentage of each species makes it possible to calculate certain phytosociological parameters such as Absolute Frequency (AF), Relative Frequency (RF), Absolute Dominance (DoA), Relative Dominance (DoR) through use the following formulae (MUELLER-DOMBOIS; ELLENBERG, 1974):

No. of squares of species i              AF of species i

AF=       ----------------------------------     RF=      ------------------------------      X 100

Total no. of squares                        Total frequency of species

Coverage percentage of species I            Absolute dominance of species i

(DoA) =  ---------------------------------  DoR=  ----------------------------------  X 100

Total sampled area                                   Total dominance of the species


For the purposes of species ordination, the Coverage Index (CI) may be used, according to MANTOVANI (1987): CI = AF + DoA

Species curve per sampled area.

Figure 3. Schematic, showing the layout of the plots for herbaceous aquatic macrophytes in the PPBio flooded and floodplain areas.

Floristic survey:

In addition to the quantitative samples obtained using the plot method, a simultaneous floristic survey will be carried out through the collection of all fertile species found within the work area, in addition to those found within the plots themselves.

Technique 2. Land-based herbs, epiphytes and hemiepiphytes

In terms of land-based herbs, epiphytes and hemiepiphytes, the width of the area sampled will be a total of 2 m, resulting in a sampled area of 0.05 ha (250 m x 2 m) per plot. The plot must be set out on either side of the centerline and after the 1 m buffer area (the area alongside the centerline), in order to facilitate researcher movement. Note: so that this plot can remain permanent, it is important that researchers do not trample through the area in order to measure trees or other types of specimens. It is therefore best to mark out the area on both sides. The demarcation line must extend out at +/- 30 cm from the ground, connecting the guide stakes that follow the demarcations set out at 10 m from the centerline. These stakes, set out every 10 m, mark out the sub-plots. Researchers working within other groups must be advised, therefore, not to walk around in this area. For epiphyte and hemiepiphyte species, the number of trunks where the species are found, up to a height of 8 m, will be counted. The 10 m sub-plot in which each ‘individual’ is present must be noted down in the same manner as that of land-based species. The number of the individual tree on which each epiphyte/hemiepiphyte was found should be noted in case the tree has already been marked within the plot, enabling host preference analysis to be carried out when the trees are identified. All individual herbaceous individuals will be counted in each of the sub-plots and, for certain species, also their measurements. Delimitation of individual species is often very difficult, due to many species being clonal. In terms of clonal species, the number of stems or rosettes (‘ramets’) will be counted, except where clusters occur, irrespective of whether the individuals are genetically different or whether, due to low density, there are species by which individuals can be distinguished. However, the entire gradient must be covered in order to determine the habitat. Permanent marking out of individual specimens for mortality, recruitment and growth monitoring purposes can also be carried out, provided that the effort required does not compromise the complete realization of the basic inventory across all plots. Marking out of individual specimens can be done with light aluminium plates and jacketed wire (telephone wire), tied to a thin wooden stake or metal skewer and located next to the plant, with the numbered plate tied to the stake. The wooden stakes are only temporary, lasting for about a year, but should be reused if the duration between censuses is short.

Coverage Measurements: Coverage measurements will be carried out using the exact plot method, but only for herbs found on land. A measuring tape is extended out along one of the sides of the plot; a very thin metal rod (e.g. a welding rod of 2 mm in diameter) being vertically positioned every 10 cm, touching the measuring tape. If any part of a herbaceous plant touches the metal rod, the species will be counted at this point. The measurements are repeated every 10 cm along the entire 250 m of the plot, resulting in a sampling intensity of 2500 points per plot. These values are used for calculating the coverage of each species per plot (number of points at which the species touches the metal rod / 2500). It is important to consider the two following points: 1) More than one species could potentially touch the rod at the same time, both of these being counted; 2) The same species can touch the rod more than once at the same point, but at different heights. In such situations, the height at which each touch occurred will be used for coverage calculation at different strata (heights), but only one touch is used for calculating the species coverage. If no plant touches the metal rod, coverage at this point is recorded as zero. Due to the inner side of the plot being more exposed to possible effects of trampling, it is recommended that the measurements be taken at the side most distant from the centerline. This is so that repetitions of these measurements can be taken in the future and trends and changes in coverage can be detected. The coverage measurements are, in general, more objective than the individual counts (due to the issue of clonal species) and should always be carried out even if counting of individuals also goes ahead. This will ensure a measure that can be compared between different areas, irrespective of the criteria used for defining individuals. It is necessary to place metal rods as vertically as possible and not interfere with them again, even if there is a plant close by, so that the coverage measurements remain as objective as possible.  Also, there is a strong tendency on the part of trainees or inexperienced personnel to try and lean or position the metal rod so that it touches a plant. It is therefore important to train researchers before carrying out real-time measurements.

Collection technique 3. Pteridophytes

Complete samples of small specimens are easy to collect by simply taking the entire plant, making sure that in the case of Herbaria samples and species with creeping stems, that a piece of this plant with two contiguous leaves, of which at least one leaf must be fertile, is collected. For plants with upright stems, these can be cut lengthwise through the middle, making sure that each piece has fertile leaves. A sterile leaf must be prepared separately, this being of special importance in cases of dimorphism. It is not possible, however, to conserve complete examples of tree ferns and other large types of pteridophytes, being important to proceed as follows in these particular situations, by collecting: a) A 35 cm piece of the petiole of the fertile leaf, removed right next to the trunk. This is important so that the basal portion is retained, sometimes being sigmoid, other times not. This basal portion can be split lengthwise down the middle, creating two examples. It being very important to conserve the scales or trichomes found at the base of the petiole. b) The lower part (35 to 40 cm) of the leaf, showing the size and position of the basal pinnae, should not to be folded at the insertion point, potentially changing its normal position. Collected leaves may be folded at the tip or in the middle if they are very big. c) The middle part of the leaf at the height of the largest pinnae; remove a piece of the rachis, leaving one complete pina only and two or three pinnae bases, either on the side of the conserved pina or the opposite side, enabling the position relative to the insertion to be viewed (whether opposite or alternate), in addition to spacing. d) The apical part of the leaf, to show whether the pina is similar to the side pinnae or whether it is different. e) The apical part from the middle of a pina, from the middle of a sterile leaf (including a piece of the rachis). It is recommended to collect the plant even if it is sterile, identification being possible even at species level. Collection of this sterile example is also important for contributing to the understanding of the geographic distribution of the family or genera to which this example belongs. Remember, never to collect a single specimen, but a good number of duplicates whenever possible. This type of vegetation is rarely found in other regions throughout Brazil, therefore being of vital importance not to eradicate a species from its habitat by removing excessive amounts of material (MORI, et al., 1989; WINDISCH, 1992).

Sampling unit for pteridophytes: a plot of 5 x 250 m.

Sampling design for pteridophytes: the sampled width will be 5m, resulting in a total sampled area of 0.125 ha (250 m x 5 m) per plot. The pteridophyte plot must therefore be established on one side, or other, of the centerline (Figure 7). It is best, therefore, to mark out a straight line demarcating this area on both sides. This demarcation line must be extended out at ± 1m from the ground, connecting the guide stakes that follow the demarcations, set out at 10 m from the centerline. The pteridophyte species will be collected (at least one sample per plot) from the 30 (thirty), 5 x 250 m plots, the respective individual specimens being counted. The epiphytes and hemiepiphytes will be sampled in the understory at a height of 3 to 4 m from the ground, without the aid of climbing equipment. The habitat of each species will be recorded (land-based, epiphyte, hemiepiphyte, rupicolous or aquatic).

Important environmental data for pteridophytes: Height, incline, soil (texture, fertility, hydric potential), tree structure, habitat (land-based, rupicolous, epiphyte, hemiepiphyte, aquatic), environment of occurrence (inner forest, forest edge, trail edges, ravines, solid land, river or stream banks or the edge, headwater or inner region of Igapó wetlands).

Collected material preservation method for higher plants: The samples will be dried in gas or electric ovens, preferably in the field or alternatively at the laboratory. Material sent to the laboratory must be soaked in 70% alcohol, thereby ensuring greater durability and avoiding the loss of leaves and reproductive parts. Araceae inflorescences and infrutescences must be soaked in glycerol (50% glycerin, 50% distilled water). All other procedures will then be carried out at the laboratory (identification, mounting, recording and sample inclusion). Unlike a number of vegetable groups that need to have their materials immediately pressed, or have specific organs fixed in preservative liquid, pteridophytes are an easy type of specimen to be collected. Exsiccates can be left for hours (days if necessary) in closed plastic bags with no holes, stored in the shade with a few splashes of water and a small amount of air to keep the material moist. The collected specimens will be deposited in trustworthy Amazon depository collections.


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