JARS v59n4 - Collection of Rhododendron Section Vireya in Sulawesi, Indonesia for Studies into their Evolutionary Relationships and Biogeography

Collection of Rhododendron Section Vireya in Sulawesi, Indonesia for Studies into their Evolutionary Relationships and Biogeography
Lyn A. Craven
Australian National Herbarium, CSIRO,
Canberra, Australia

Gillian K. Brown
Australian National Herbarium, CSIRO, Canberra, Australia, and
The University of Melbourne, Melbourne, Australia

Lina S. Juswara
Herbarium Bogoriense, Botany Division,
Research Centre for Biology, Indonesian Institute of Sciences, Bogor, Indonesia.
(Present address: The Ohio State University, Columbus, Ohio, USA)

Summary

Several mountains in Sulawesi, Indonesia, were explored to collect study materials of Rhododendron section Vireya as part of a project investigating the evolutionary relationships and biogeography of the group. Leaf tissue for DNA sequence studies and herbarium specimens for morphological observations and for voucher specimens were collected from at least thirteen species of Vireya .

Project aims

The objective of the project was to collect materials of species of Rhododendron section Vireya that occur in Sulawesi, Indonesia, for a larger project being undertaken by Gillian Brown, in Canberra, Australia, for her Ph.D. investigations into the phylogenetic relationships and biogeography of Rhododendron section Vireya . Significant contributions are being made in understanding evolutionary relationships and biogeographical patterns using DNA sequence data and this project aimed to explore the relationships of Vireya species using these techniques. The current classification of section Vireya (Chamberlain et al. 1996) is based upon the work of Sleumer, notably his account of the genus in Flora Malesiana (Sleumer 1966). Sleumer's classification was based upon morphological features, the traditional source of information used by taxonomists to devise plant and animal classifications. We wished to test the classification using data from a different source, and molecular sequence data is ideal for this purpose.

Relatively more plant exploration has occurred in western (esp. the Malay Peninsula and Borneo) and eastern (Philippines and New Guinea) Malesia than in the central islands of Malesia, which are incompletely known. This discrepancy also applies to Rhododendron despite it usually being a distinctive and hence commonly collected component of the vegetation. One of the largest islands in central Malesia, Sulawesi, has several high mountain ranges, and has potentially acted as a major stepping-stone in the dispersal of Vireya species through the Malesian archipelago. Consequently, we wished to ensure that Vireya species from Sulawesi were well represented in the r

esearch project.

Fresh leaf material is optimal for the isolation of DNA, although DNA can be extracted from herbarium specimens. However, the older the herbarium specimen the more likely the DNA will have degraded and is therefore unsuitable for use in studies such as this. In particular, field preservation techniques of herbarium collections and subsequent storage conditions can adversely affect DNA viability, making it almost impossible to obtain informative DNA from the specimens. The majority of the available herbarium specimens of rhododendrons from Sulawesi were collected many years ago and, therefore, the DNA is too degraded, making it necessary to undertake field work to obtain suitable study materials.

Team

The field party consisted of Lyn Craven and Gillian Brown, from the Australian National Herbarium, CSIRO Plant Industry, and Lina Juswara, from Herbarium Bogoriense, LIPI, Indonesia. Lyn is a botanist in CSIRO and Gillian was a postgraduate student there (with university affiliation with the University of Melbourne); Lina is a botanist with the Herbarium Bogoriense, LIPI, Bogor. At the last two field locations visited, the team was accompanied by Ramadhanil, a lecturer in plant sciences at Universitas Tadulako, Palu, Central Sulawesi.

Field work

An application to conduct the collecting work was lodged with the Bureau of S & T Cooperation, LIPI (Indonesian Institute of Sciences), in Jakarta in 2001 and once their permission had been given, several months later, travel dates and logistical matters were addressed. The sites identified for exploration were selected on the basis of prior knowledge that they possessed moderate to high levels of species diversity in Rhododendron . The final sites selected were Gunung Rantemario, in the province of South Sulawesi, and Gunung Sojol and Lore Lindu National Park both in Central Sulawesi.

In June 2002, Lyn and Gillian travelled to the host institution Herbarium Bogoriense in Bogor, West Java, where they were met by Lina, the counterpart officer. Lina's local knowledge was essential throughout the field trip, especially in obtaining all the necessary documentation and communicating with people in the local villages visited. With an imperfect knowledge of Indonesian, this field trip would not have been possible.

From the 24-28th June, the permits and travel documents were obtained. These activities usually occupied the mornings and we made use of any time available in the afternoons to purchase field equipment in Bogor that could not be obtained in Sulawesi.

On the 29th of June we flew to Makassar (the original name for Ujung Pandang), the provincial capital of South Sulawesi, and became familiar with the city before presenting the appropriate documents at the provincial administration and forestry offices, where we received further documentation for district offices.

We travelled by hire car to Parepare on the 2nd of July. There we commenced the administrative process whereby the several levels of provincial government acknowledged our wishes to do research in South Sulawesi and provided us with the necessary documentation. We stayed in the town of Rantepao in the Tana Toraja district while the paperwork was being done. Once that was completed, we drove to Baraka where we hired a 4WD truck to take us on to Pasongken. There we stayed the night at the home of the Kepala Dusun (kampung head man) at Buntu Mondeng.

The following day we travelled by horse along the road and then bridle path to Karangan, the kampung at the foot of G. Rantemario, where we stayed at the home of the Kepala Dusun of Karangan. That afternoon we negotiated for porters to carry our camping and collecting gear and arranged to start for G. Rantemario in the morning. On the morning of the 6th of July, we walked to Pos 2 where in the afternoon we collected along the path above the campsite. We spent a difficult, sleepless night on a narrow, slightly sloping outwards rock ledge above a very noisy waterfall. (Hank Helm and John Farbarik experienced the same conditions. We later discovered that there is another campsite further along - at Pos 5 we think it was - and we recommend that future visitors plan to stay there. "Waterfall Lodge" is just too scary.)

After our restless night we hiked to Pos 7, at just over 3,100 m, and set up camp. Along the way, we made our first collections of Rhododendron . (Between Pos 2 and 3: R. sp. [perhaps R. rhodopus ]; between 4 & 5: R. malayanum var. malayanum ; between 5 & 6: R. lagunculicarpum , R. sp. [cf. zollingeri ]; at 7: R. eymae .) More collections of Rhododendron were made the next day on route to the summit ( R. buxifolium , R. eymae , R. nanophyton ). We broke camp the next day and returned to Karangan, collecting some more Rhododendron along the way. (Just below Pos 7: R. pudorinum ; between 4 & 3: R. celebicum ). The following day we retraced our steps and travelled back to Pasongken by horse, to Baraka by 4WD truck, and to Rantepao by hire car, arriving without incident in Rantepao on the 10th of July.

After a well needed day of rest and washing, we conducted a day trip to Batutumonga, a kampung well up on G. Sesean. We collected R. rhodopus along the road but due to blistered feet and heavy colds, legacies from G. Rantemario, we did not attempt to walk further up the mountain. After returning from Batutumonga in the afternoon, we came across the stunning R. vanvuurenii on the southern outskirts of Rantepao.

On the 13th of July we headed for Palu in Central Sulawesi, by hire car. On the way we crossed two ranges (between Rantepao and Palopo, and the range S of Danau Poso) that would be worth exploring for Rhododendron , but due to limited time, we could not spend time exploring. Just south of Pendolo we were stopped at a large police roadblock, but were allowed to proceed (6 check points in all, the police were carrying arms at the first few). We were informed that there had been a bomb attack on a bus in Poso the previous day, reportedly killing five people. That night we stayed at Pendolo while we reviewed the situation. We decided it was best that we return to Makassar and travel to Palu by air, the major reason being that we did not wish our driver to have to travel alone on his return journey from Palu. The following day we drove for twelve hours down the east coast and then crossed to Parepare, thence to Makassar. We spent the next day in Makassar air freighting herbarium collections to Bogor and booking flights to Palu for the following day. On the 16th we flew to Palu where we were met by Pak Ramadhanil, who was to accompany us to G. Sojol and Lore Lindu. Ramadhanil assisted greatly with local knowledge, arranging vehicles, etc.

We visited the appropriate government offices in Palu (provincial, district, forestry) and were informed that we were required by forestry to take one of their staff with us to G. Sojol, our first destination. That afternoon we shopped for supplies and met our newest companion from the forestry office.

On the 18th of July we travelled by chartered minibus to Siboang, on the coast immediately west of G. Sojol, where we stayed the night at the Kepala Dusun's house. The next day we went by bullock cart to Maros where we luckily encountered the Kepala Dusun from Sipatoh, the last kampung before G. Sojol. He sent back porters to carry our gear the half hour's walk to Sipatoh and guide us to his village. In the Kepala Dusun's house we negotiated for seven porters to take us onto G. Sojol, and they also required us to pick up two others along the way as guides; these two were members of an animist community and were necessary to placate the several deities on the mountain.

We left Sipatoh the next day and walked up to a peak c. 1,100 m high where we camped at a site near a small spring. The following day we walked for 8.5 hours to a sungai (stream) and followed it along to a better campsite near a tiny rivulet. The porters were extremely unhappy about proceeding further up the mountain without the animist guides but this was going to take another day to organise, as they lived across the ridge and not in the direction of the summit. As we did not have this time to spare, we made the decision to abort this sector of our travels as time was against us. We returned slowly to our first camp. On our way back, we sheltered from a heavy shower in some garden huts, and trudged in the mud before setting up camp on a high ridge about halfway between the sungai and our first camp. The next day we returned to our first campsite on G. Sojol. On the way, we collected two species of Rhododendron ( R. sp., R. malayanum var. malayanum ). We spent the night at this campsite before returning to Sipatoh, collecting another Rhododendron (perhaps R. rhodopus ) along the path.

After a night in the village of Sipatoh we again retraced our steps, farewelling our newfound friends and hosts. We walked to Maros, travelled by bullock cart from there back to Siboang, and then returned to Palu by minibus, arriving well after dark. The next day, the 26th of July, we attended to the specimens from G. Sojol and visited Ramadhanil's herbarium, the Herbarium Celebense, at Universitas Tadulako. From discussions with Ramadhanil and Christian Schulze, an entomologist from the German-funded conservation project, STORMA, based in Palu, we decided to conduct a day trip to Lore Lindu National Park, where they had previously seen three species of Rhododendron in flower. Somewhat to our surprise, we had an extremely productive visit to Lore Lindu National Park, where we collected seven species of section Vireya : one Vireya species at each of the first two stops and five along the upper reaches of a road on one of the mid-height northern peaks in Lore Lindu (at Puncak Padeha: R. vanvuurenii ; at Danau Kalimpaa: R. sp.; at Gunung Rorekatimbu: R. ?alternans , R. malayanum var. malayanum , R. radians , R. rhodopus vel aff., R. zollingeri ).

Our day trip to Lore Lindu was also our last day in Sulawesi. We flew from Palu to Makassar, and then onto Jakarta on the 28th of July, returning to Bogor that evening by commercial charter vehicle. We had several days in Bogor, where we were able to attend to the first of our dried specimens at the LIPI herbarium, packed our materials ready for transport back to Australia and also visit the lower montane botanic garden at Cibodas in West Java (Kebun Raya Indonesia Cibodas), where there were three vireyas in cultivation, R. album , R. javanicum and R. macgregoriae . On the 2nd of August we arrived back in Canberra, although our specimens and living collections had to remain in Sydney over the weekend for Australian quarantine procedures. (On the Monday, the CSIRO quarantine officer arranged for all the materials to be sent to Canberra by overnight carrier. The materials were received by us the following day.)

Other than difficulties created by the community conflict in Poso, Central Sulawesi, that prevented road travel to the second set of field sites, no significant problems were encountered by the field party and it was a very successful field trip.

Scientific results

At least thirteen species of section Vireya were collected for herbarium and molecular studies, and living material was collected of about twelve of these. The exact number of species collected will not be known until plants are grown on to flowering, at which time identification will be possible. The rhododendrons collected are listed in Appendix 1.

Assessing the taxonomic status of several of the accessions has proved difficult. Introgression apparently has led to the development of populations that cannot be unambiguously assigned to species in Sleumer's 1966 account of the section in Flora Malesiana . Given that relatively few specimens from Sulawesi were available to Sleumer, it may well be that the circumscription of some of the species he recognised may need to be amended. It is possible that our #129 may warrant taxonomic recognition at some level as, while it clearly is very similar to R. rhodopus , it differs in several respects from that species.

DNA was successfully extracted from the samples collected and utilised in Gillian's studies, which have now been completed. The key findings will be presented in another article for the ARS Journal once the scientific papers emanating from her research have been published.

Where it was encountered, seed was collected for growing plants in Canberra for further study. When the seed was in sufficient quantity, some was sent to expert growers in the USA and New Zealand. Unfortunately, much of the seed collected was extracted from long-dehisced capsules and was subsequently found to be inviable. The cuttings were taken to Canberra for quarantining and propagation. Later, when plants are established in Canberra to a sufficient size, material of all surviving accessions will be sent to Kebun Raya Indonesia and also to the USA and New Zealand.

Herbarium specimens of the Rhododendron collections have been deposited in the herbaria at Bogor and Canberra, as well as in kindred institutions. Other species of interest were collected by the field party when time permitted and specimens of these were similarly deposited in relevant herbaria; other genera collected include Syzygium , Clematis , Ficus , Rubus , etc.

Appendix 1
List of Rhododendrons collected on Sulawesi, July 2002

Herbarium specimens were made for each collection. If living material was collected it is indicated with "L".

Gunung Rantemario
20    sp. (ser. Javanica , rhodopus or similar - need flowers for identification) L
26 malayanum var. malayanum
27 malayanum var. malayanum L
30 lagunculicarpum
31    sp. (subsect. Albovireya , cf. zollingeri - need flowers for identification) L
34 lagunculicarpum L
41 eymae
43 eymae L
44    ? lagunculicarpum hybrid L
45 pseudobuxifolium
46 nanophyton
49 nanophyton
52 pseudobuxifolium
76 pudorinum (but need flowers for confirmation) L
77 celebicum L

Gunung Sesean
78 rhodopus L

Rantepao
79 vanvuurenii

Gunung Sojol
114    sp. (subsect. Solenovireya - need flowers for identification) L
115 malayanum var. malayanum
sine numero sp. (ser. Javanica , rhodopus or similar, need flowers for identification) L

Lore Lindu National Park
123 vanvuurenii L
124    sp. (need flowers for identification) L
125 zollingeri L
126 malayanum var. malayanum L
127    ? alternans L
128 radians (long corollas, 8.5 cm when dried) L
129 rhodopus (perhaps worth taxonomic recognition; inter alia differs in leaf features, long corollas (8 cm when dried], fragrance) L

Acknowledgments
The Research Foundation of the American Rhododendron Society is thanked for their support of this project. Financial support was also provided by CSIRO Plant Industry and CSIRO International Scientific Liaison. Our companion at G. Sojol and Lore Lindu NP, Pak Ramadhanil, facilitated logistical arrangements and guided us to Rhododendron-rich areas in Lore Lindu NP; he also was a first class field companion. John Farbarik, Hank Helm, David Binney, Mary Mendum and George Argent gave advice on access routes to G. Rantemario and G. Sojol. John and Hank generously provided detailed notes from their own expedition. Mary and George introduced us to "leech socks", valuable devices to help prevent these undesirable animals from gaining access to one's legs.

The Bureau of S & T Cooperation, LIPI, is thanked for approving the research project. The Director and staff of the Herbarium Bogoriense are thanked for their willing and effective assistance in planning and conducting the field work and subsequently in dealing with the collected materials.

Finally, the Kepala Dusuns of Pasongken, Karangan, Siboang and Pasongken, Karangan, Siboang and Sipatoh kampungs in Sulawesia are thanked for their hospitality. We especially thank the Kepala of Karangan and of Sipatoh and those people of their kampungs who carried our camping and collecting equipment and guided us through their forests. Although our time in the field with them was short, we have the feeling that genuine friendships developed at the level of person to person.

References
1. Chamberlain, D., Hyam, R., Argent, G., Fairweather, G. and Walter, K.S. 1996. The genus Rhododendron: its classification & synonymy . Royal Botanic Garden Edinburgh: Edinburgh.
2. Sleumer, H. 1966. Rhododendron . Pp. 474-668. In: C.G.G.J. van Steenis (gen. ed.), Flora Malesiana ser. I, vol. 6, Wolters-Noordhoff Groningen.

Author biographical notes and addresses:
Lyn Craven is a plant systematist with the Australian National Herbarium in Canberra, Australia. His research is focused on Myrtaceae, the myrtle family, and Malvaceae, the hibiscus and mallow family, although he has also published on several other families. Lyn has collected in Australia, New Guinea, Fiji and New Caledonia, in addition to the present work in Sulawesi. One of his hobbies is horticulture and Lyn has been growing Vireya rhododendron species for more than forty years. He has been a member of the ARS for nearly this long.
Gillian Brown is a postdoctoral research fellow in the School of Botany at the University of Melbourne, Melbourne, Australia. She recently completed her Ph.D. dissertation on the phylogeny and biogeography of Rhododendron section Vireya and is presently writing up these results for publication. Gillian's interest in Vireya rhododendrons and the history of the Malesian Archipelago remains strong, even though her current research focuses on the relationships of the Australian species of Acacia .
Lina Juswara is a plant systematist with the Herbarium Bogoriense in Bogor, Indonesia. Her research is focused on Orchidaceae , a family that is very diverse and species-rich in Indonesia. Lina presently is studying a group of Indo-Pacific ground orchids for her Ph.D. at the Ohio State University, Columbus, USA.