MANAGEMENT OF Pyrrosia Mirb. ( POLYPODIACEAE ) FERN SPECIMENS IN HERBARIUM DEPOKENSIS (UIDEP) UNIVERSITAS INDONESIA

The herbarium harbors collections of preserved plants used for scientific purposes. It is important to keep information about plants in a valid herbarium collection. This study aims to establish a valid collection and database of Pyrrosia stored in Herbarium Depokensis (UIDEP) through herbarium management. The management process includes taxonomic validation, specimen revitalization


INTRODUCTION
A herbarium is a collection of preserved plants stored in a room with specific environmental conditions, such as temperature and humidity.Herbariums are usually managed by scientific institutions that are responsible for storing and administering their use (Bridson & Forman, 1992;Bromberg, 2020).The herbarium is a repository for reference material, a means for identifying species, a source of confirmed taxa names, and a comprehensive data bank.Aside from its traditional role, herbarium nowadays has broader purposes.Digital and traditional collections have been used to document plant ecology such as the range of rare species, its declining patterns, or the spread of invasive plants.Ecological botanists compared specimens from different times and places using herbarium collections (López & Sassone, 2019).The herbarium data is digitized to make it more widely accessible.The database could also be sorted in various ways to fulfill many practical needs outside of the traditional alphabetical sorting of the taxa.For example, the available taxa can be sorted according to the area of distribution or its genetic relationship to other taxa, to further understand a certain taxon in both matters.Those data are important as a source of information related to taxonomic information on plant specimens.A researcher can use the herbarium collection stored as a reference to identify other specimens that are not yet recognized or find out the distribution of taxa by looking at the description of the location of the collection on the label.Therefore, the herbarium is considered an asset and a wealth of knowledge because it stores taxonomic and ecological data and represents plant diversity (Besnard et al., 2018;Yaman et al., 2022).
Some local herbarium units are managed by schools or universities to act as learning facility.One of them is Herbarium Depokensis (UIDEP) in the Department of Biology, Faculty of Mathematics and Natural Sciences Universitas Indonesia.UIDEP stores a collection of herbarium specimens from various locations in Indonesia, but mostly from UI's Green Spaces (RTHK).The specimens are collected from biodiversity exploration conducted by the students, or outcomes from practice classes.At least 400 of the specimens stored in UIDEP are Pteridophytes.One of the notable genera in Pteridophyta is Pyrrosia, which was first described by Mirbel (1803) (Hovenkamp & Wessendorp, 1986).Pyrrosia generally have thick and fleshy simple leaves.Pyrrosia leaves are usually dimorphic (different leaf shapes between sterile and fertile leaves) (Sofiyanti et al., 2015).Pyrrosia species is used as an ornamental and medicinal plant.Malays and Filipinos used to use Pyrrosia leaves as an external medicine.P. piloselloides leaf juice is reported to be used by Indonesian people as a cough medicine, constipation, and gonorrhea (Fatimah, 2009;Natalia, 2012).
The collection of Pyrrosia specimens stored in UIDEP has not been well-curated.The problems regarding the specimens themselves were incomplete diagnostic characters of the fern, mainly caused by physical damage while storing or insufficient herbarium preservation method.In addition, there is a large part of the collection with old taxonomic information that has not been updated.This includes the species name in some of the specimen labels being Drymoglossum (Pyrrosia's synonym).To maximize its function, herbarium collections must be correctly maintained which can be called Herbarium Management (Harris & Marsico, 2017).
This study focuses on two kinds of work in herbarium.One is about the taxonomy aspect of herbarium collections stored in UIDEP, which includes its accuracy in naming the plant specimen and how well the taxa were grouped according to the latest taxonomy classifications.The other aspect is the technical handling of those herbaria itself.That comprises how well the specimens were stored in the unit, and also the standard procedure of collecting and arranging a plant on herbarium paper.The objective of this study is to carry out Herbarium Management through a revitalization of all specimens which includes physical condition analysis, evaluation of species naming, and revitalization of each Pyrrosia specimen in UIDEP before digitizing the herbarium in an online database for easier access.

MATERIALS AND METHODS
The validation and specimen digitization activities were carried out in Herbarium Depokensis (UIDEP), Biota Collection Room Department of Biology, FMIPA, Universitas Indonesia, Depok.The herbarium sheets that will be examined are all specimens named Pyrrosia spp.and Drymoglossum spp. on its label, a total of 59 specimens labeled as P. piloselloides, P. lanceolata, P. longifolia, P. nummularifolia, D. piloselloides, Drymoglossum sp., and Pyrrosia sp.When observing herbarium specimens for species validation, a Nikon Stereo Microscope SMZ745 and an Olympus long-arm microscope were used.A computer device is used to input data from the validation stages to the final database.When remounting the specimen, sewing thread, paper tape, scissors, glue, stationery, and isolation tape are needed.Revision labels are also used to mark the revised taxa as needed.Finally, specimen documentation for digitization using the Canon EOS 500D DSLR camera that was attached to a Canon Copy Stand.
Herbarium management is carried out in three main revitalization steps (Natural Herbarium of Cultivated Plants, 2019), which include taxonomic nomenclature validation, specimen repairment, and digitizing all specimens (including documentation and databasing).First, all specimens identified as Pyrrosia spp.and Drymoglossum spp.were re-grouped based on morphological characters' similarities between specimens.The method for morphometry was done through detailed morphological examination.The main characters of each group were compared with references, to confirm that the specimen is the species name that is written on the label.In this study, a couple of reference sources were used such as the Pyrrosia monograph by Hovenkamp and Wessendorp (1986), Hovenkamp et al. (1998), and PPG 2016 to validate specimens' nomenclature.Herbarium specimens were compared with each other's characteristics and determined whether or not they belonged to the same taxa group.The validated specimens are then ready for the repairment process.Repairment of all specimens has been made for 2 aspects, the physical condition of the herbarium sheets and information validation written in the label.Each herbarium sheet was carefully observed, and problems appeared and were identified.Both specimens' repairment and specimen' label checking were done based on herbarium references (Christenhusz & Chase, 2014).Digitization was carried out for all valid herbarium data resulting from previous work stages.The process was done in two steps, specimen documentation and data uploading to the website.Pictures were taken for each specimen with a DSLR camera that was mounted on a Copy Stand, the use of a photo lamp as a modification of lighting was made to ensure that the photos looked as close as the actual specimen.All valid data in the herbarium label were transferred into an Excel spreadsheet.Both the pictures and specimens' data were uploaded to Herbarium UIDEP online database (www.ruangkoleksibiotaui.id).

Nomenclature Validation
All 59 herbarium sheets named Pyrrosia spp.and Drymoglossum spp. on their existing label before the validation process were then confirmed as Pyrrosia.The diagnostic character for this genus is the stellate hairs found on the surface of the lamina.Detailed morphological examination of those specimens resulted in three taxa of Pyrrosia (P.piloselloides, P. lanceolata, and P. longifolia).There was some nomenclature difference between the existent label and validation results (Table 1), which then were updated to the latter.Nomenclature changes were made possible by detailed morphological examination of several key characters of P. piloselloides, P. lanceolata, and P. longifolia (Table 2).Validation of all specimens as Pyrrosia was indicated by long-creeping rhizomes, simple leaves, and entire leaf margins.The diagnostic character of the genus Pyrrosia found was the presence of stellate hairs on the surface of the lamina.The stellate hairs have acicular rays and were found to be denser on the abaxial part of the leaf than on the adaxial.The presence of stellate hairs distinguishes Pyrrosia from other genera of ferns which also have thick leaves and entire leaf margins such as Lemmaphyllum (Donk, 1954).
To validate the species name within Pyrrosia diagnostics were used including macromorphological and micromorphological characters.The macromorphological characters observed were rhizome shape and size, distance between phyllopods, and leaf shape (including apex, base, and stalk).In general, the two species can be distinguished by the apex and/or basal of the sterile lamina.In P. lanceolata the apex and/or basal part is more tapered and the widest part of the lamina is at the bottom or top of the middle of the leaf (lanceolate).In contrast, P. piloselloides has an overall elliptical leaf shape.P. longifolia has a leaf shape that is quite different from the other two species because of its strap-shaped leaf.The length of the leaves of P. longifolia can reach 20 cm or more (in specimens, the longest found is 54 cm).
Micromorphological diagnostic characters used to distinguish the three groups of Pyrrosia specimen collections in Herbarium UIDEP were rhizome scales and sori.Rhizome scales are found along the surface of the rhizome to the phyllopod of the Pyrrosia.The results of rhizome scales observation with a microscope showed that there were variations in the shape of the scales between species (Figure 1).All of the rhizome scales observed in the Pyrrosia specimens had the same attachment, which is peltate (attached to the center of the scales).
Based on Figure 1, the differences between species were indicated by the shape of the scales and the presence/absence of cilia at the margins of the scales.The shape of the rhizome scales in Figure (1a) was found in the P. piloselloides specimens.The margin of the rhizome scales was ciliate and did not have a long acumen (pointed angle).The size of the scales is also the smallest compared to the other two groups, namely ±1 mm long and 0.8 mm wide.In P. lanceolata, peltate scales were found (Figure 1b).The margins of the scales were also ciliate but there is an acumen structure that looks elongated.The length of the scales of P. lanceolata can reach 1.5 to 3 mm while the width is 0.3-1.2mm.The scale form (c) was found in the P. longifolia.Form c looks most distinctly different because the scale margins were flat without cilia (entire) and also have acumen.The size of the scales observed in P. longifolia specimen was ±1-3.2 × 0.6-1.5 mm.

Physical Examination and Specimen Handling
Herbarium sheets examination on all specimens identified types of damage and inaccurate information in the labels as shown in Table 3.Most of the problems occurred on the plant itself, whether it is from physical damage or not enough representation of preserved plant parts on the herbarium sheet.Damages found include the crumbled lamina, decays caused by insects, etc.The second most problems were incorrect nomenclature in 51 sheets.Incorrect names were due to missed identification and outdated classification.The other problem is incomplete herbarium in Pyrrosia specimens, mainly lack of fertile lamina of the fern.Apart from carrying out physical improvements, additions to the collection were made by collecting fresh specimens from the field.Fresh sampling from the field was carried out to add to the collection and complete the lack of character data.New Pyrrosia samples were also collected to make a voucher herbarium and complete the data of existing specimens without the fertile lamina (Table 4).Incorrect nomenclature is the biggest problem in Pyrrosia specimen collections.Identification errors or nomenclature updates on the labels were corrected by adding revision labels.The label contains the correct nomenclature/taxa description and is mounted to the top of the existing herbarium label (Figure 4).

Digitization of the Specimen
The valid herbaria resulting from the revitalization process (both specimen and label) are documented to an online database.All specimen sheets were digitally documented and resulted in Problems identified in Pyrrosia herbarium collection are samples detached from duplex, lack of fertile lamina, damages to the fertile lamina, incorrect nomenclature, incomplete label notes, and insects found on the specimen.Every problem is resolved with appropriate treatment.
Damages on the lamina were resolved by removing the damaged part but still paying attention to its representative plant parts, so it stays complete and not reducing any information on herbarium specimens.Poor mounting causes sample specimen to detach is solved by rearrangement of the specimen on the duplex paper.Existing herbarium specimens that have damage on the plant parts can be repaired with the same equipment as the general mounting process in making the herbarium.Specimens were arranged above the duplex paper and remounted if the old glue/tape had loosened and/or the specimen position was disorganized.
Incomplete label notes found in specimens were mostly due to a lack of detailed informations about the samples.In that case, additional notes regarding the morphological features of the plant were added.The data was taken from observations throughout the nomenclature validation process so that the future user will be informed about the specimen's characteristics, especially if its nomenclature were updated.
A good herbarium relies on the complete and accurate information stated on its label.The label should have information on the specimen's scientific name, vernacular name, date, and location of collection, collector's name, and habitat/ecological data.It should also include supplementary data such as notes on colors, plant habits, and other features that may be important for identification but cannot be shown well as a preserved specimen.Identification errors or nomenclature updates on the labels were corrected by adding revision labels.The label contains the correct nomenclature/taxa description and is mounted to the top of the existing herbarium label (Figure 4).Based on the latest Pteridophyta classification PPG 2016, it is stated that the Drymoglossum and Saxioglossum groups are included in the genus Pyrrosia (PPG 2016: 590).
To maintain the specimens' physical condition after revitalization, a standard procedure of all herbarium works was established.This includes standard storing temperature and humidity, regular examination for pests or other potential problems, and new specimen guidelines for future collections.The references used during taxonomic validation were also used to determine what plant parts need to be present on a Pyrrosia herbarium specimen.Collecting ferns for a herbarium is not just about taking the plant and preserving it, but planning so the information within can be observed and studied even after a long time.When collecting and preserving the plant for herbarium making, it is important to include some of the key characters to identify the plant.So that the plant's morphological characteristics could be representatively displayed on a herbarium sheet.
Based on those physical examination results, we advise some specific techniques in handling Pyrrosia specimens.First, make sure to take complete note of the fern habit before collecting.Pyrrosia species characteristics vary in many things, some are no longer visible after preservation, it is best to write them down first.Lastly, make sure to collect all parts of Pyrrosia that are crucial for identification such as rhizome and fertile lamina.

Digitization of the Specimen
The valid herbaria resulting from the revitalization process (both specimen and label) is documented in an online database (Vissers et al., 2017).The data regarding specimen identity, location, and taxonomy information were then uploaded to the Herbarium UIDEP website.It can be accessed at www.ruangkoleksibiotaui.id.(Additional pictures were provided to give more information about the said specimen to the viewers.After the herbarium data is entered on the input page, the specimen information cannot be directly visible publicly on the website. However, the uploaded data is entered in the admin menu for approval (Figure 5).The approval process by the admin is needed to protect the database from arbitrary specimen uploads.After the herbarium data entry is approved, the input results will appear publicly and can be accessed by entering the appropriate keywords on the search page (Figure 6).Herbarium data can be searched by family name, genus, species, and local name.ruangkoleksibiotaui.id)

CONCLUSION
The management has been carried out on the herbarium specimens which has resulted in 3 validated species of Pyrrosia in the Herbarium Depokensis (UIDEP) collection, namely Pyrrosia piloselloides, Pyrrosia lanceolata, and Pyrrosia longifolia.The three species can be distinguished by the morphology of the rhizome, lamina, and soral position.Nomenclature validation and specimen revitalization resulted in a collection of valid Pyrrosia specimens.Digitization of valid Pyrrosia specimen collection produced an online database that can be accessed and used by the public.
From the results of this study, it can be suggested to periodically check the results of identification or naming of specimen collections, check the physical condition of specimens, conduct collections, and build a database system.

Figure 1 .
Figure 1.Rhizome scales on 3 species of Pyrrosia.Pyrrosia piloselloides (a), Pyrrosia lanceolata (b), and Pyrrosia longifolia (c) In addition to rhizome scales, another micromorphological character observed to distinguish Pyrrosia species in Herbarium DEP was its sori arrangement.The position of the sori on the fertile lamina was observed as shown in Figure 2. The sori in Figure (a) was found in the P. piloselloides, it could be seen that the sori were arranged longitudinally at the margin.The sori arrangement in Figure (b) was found in P. lanceolata.Sori rounded, arranged in 2 rows on the abaxial surface of the leaf lamina between the midrib and margin.Sori looks a bit sunken and covered in thick stellate hairs.Sori in Figure (c) was found in the P. longifolia.Sori rounded and scattered on the abaxial surface of the lamina but only found around the apex to before the middle of the leaf.No sori were found around the leaf basal area even on very long (>50 cm) lamina.

Figure 5 .Figure 6 .
Figure 5.The input results are shown on the website admin menu (Source: ruangkoleksibiotaui.id)

Table 1 .
Comparison of Pyrrosia species naming in Herbarium UIDEP before and after the validation process (numbers refer to the amount of specimen sheets)

Table 3 .
Problems regarding the physical condition of the herbarium and repairment efforts

Table 4 .
Samples collected in the field