Ploidy Level of Endangered-Endemic Orchid Vanda Sumatrana Schltr Regenerated Plantlets
Dyah Retno Wulandari 1, a), Tri Muji Ermayanti 1,b), Erwin Al Hafiizh 1,c), Dwi Murti Puspitaningtyas 2,d), Suluh Normasiwi 3,e), and Syarifin Firdaus 4,f)

1(Research Center for Genetic Engineering-National Research and Innovation Agency: National Integrated Center for Genomic, Tropical Biodiversity and Environment Jl. Raya Jakarta-Bogor No.Km. 46, Cibinong, Kec. Cibinong, Kabupaten Bogor, Jawa Barat 16911).
2(Research Center for Plant Conservation, Botanic Gardens, and Forestry: Jl. Ir. H. Juanda No.13, Paledang, Kecamatan Bogor Tengah, Kota Bogor, Jawa Barat 16122)
3(Research Center for Plant Conservation, Botanic Gardens, and Forestry:Jl. Kebun Raya Cibodas, Sindangjaya, Kec. Cipanas, Kabupaten Cianjur, Jawa Barat 43253)
4(Orchid Bloem and Farm-Malang East Java Indonesia: Jl. Satsuitubun, Greenliving Residence D37 Malang, 65149).

Abstract

Vanda Sumatrana is an endangered-endemic Indonesian orchid according to Ministry of Environment and Forestry, Indonesia, Regulation No.106 of 2018. Plant tissue culture is a suitable technique for in vitro propagation and conservation. The ploidy level of plantlet needs to be investigated to observe genetic diversity or stability. This study aimed to analyze the ploidy level of 10-week-old regenerated plantlets of V. sumatrana compared to their mother stock plant that has been stored on in vitro culture for 3 years. Observations were conducted on 70 regenerated-plantlets from 24 parent plants. Analysis of the ploidy level was carried out using a flow cytometer. The results showed that 70.83% parental shoots or were diploids and 29.16% were tetraploids, 72.46 % regenerated shoots were diploids and 27.54% were tetraploids. Mean value of PI ranged from 107.472 to 205.162 for diploid shoots, and ranged from 258.525 to 399.255 for tetraploid shoots. This indicated endopolyploidy occurred between regenerants and their parent plants after a long period of cultures. Our finding also showed endoreduplication phenomena in this orchid species, indicated by multiple peaks of flow cytometry histograms.

Keywords: flow cytometry- endopolyploidy- endoreduplication- Vanda sumatrana- plantlet

Topic: Biotechnology