KEANEKARAGAMAN Azolla spp. DI DAERAH ALIRAN SUNGAI PROGO DAN OPAK BERDASARKAN KARAKTER MORFOLOGIS DAN INTER SIMPLE SEQUENCE REPEAT

Nafi'ah Khoirunnisa', Prof. Dr. Budi Setiadi Daryono, M.Agr.Sc.; Abdul Razaq Chasani, S.Si., MSi., Ph.D.

2025 | Tesis | S2 Biologi

Azolla adalah genus tumbuhan paku air yang terdiri dari tujuh spesies di dunia, dan hanya tiga spesies yang ditemukan di Indonesia. Analisis taksonomis dan keanekaragaman Azolla spp. di Jawa Tengah dan D.I Yogyakarta masih belum banyak dikaji. Penelitian ini bertujuan untuk mengeksplorasi keanekaragaman dan hubungan kekerabatan Azolla spp. di Daerah Aliran Sungai (DAS) Progo dan Opak, menggunakan 20 karakter morfologis dan sepuluh primer Inter-Simple Sequence Repeat (ISSR). Azolla spp. dikumpulkan secara purposive sampling pada empat belas titik lokasi. Analisis morfologi berupa identifikasi, similaritas, dan klastering dilakukan menggunakan MVSP dan Minitab. Analisis molekuler dilakukan melalui amplifikasi PCR-ISSR, dilanjutkan perhitungan nilai PIC, RP, dan variasi genetik. Dendrogram dan PCoA dikonstruksi menggunakan MVSP dan GeneAlex. Total 14 sampel ditemukan dengan sepuluh sampel dari Daerah Aliran Sungai Progo dan empat dari Daerah Aliran Sungai Opak. Karakter morfologis yang sangat berkontribusi terhadap perbedaan antara sampel adalah bentuk percabangan, keberadaan stomata ventral, warna lobus ventral, panjang lobus ventral, dan bentuk lobus dorsal. Keanekragaman spesies tergolong rendah dengan seluruh sampel teridentifikasi sebagai Azolla cristata. Di antara sepuluh primer Inter-Simple Sequence Repeats (ISSR), UBC 814 dan UBC 841 menunjukkan nilai PIC (0,29; 0,28) dan RP (19,71; 23,29) yang tinggi. Seluruh primer yang digunakan efektif dalam menganalisis variasi genetik dengan rata-rata tingkat polimorfisme sebesar 85,27%. Analisis Unweighted Pair Group Method mengelompokkan sampel ke dalam dua kluster berdasarkan sifat morfologis dan genetik mereka. Indeks variasi genetik cukup tinggi di kedua populasi, dengan variasi gen Nei (He) = 0,230 dan indeks informasi Shannon (I) = 0,342. AMOVA mengungkapkan bahwa 93?ri total variasi genetik ditemukan dalam populasi, sementara hanya 7% antar populasi. 

Azolla is a genus of water ferns that consists of seven species worldwide, with only three species found in Indonesia. Studies on the taxonomic analysis and diversity of Azolla spp. in Central Java and D.I. Yogyakarta have not been extensive enough. This study aims to explore the diversity and phenetic relationships of Azolla spp. in the Progo and Opak River Basins, using 20 morphological characters and ten Inter-Simple Sequence Repeat (ISSR) primers. Azolla spp. were collected through purposive sampling at fourteen locations. Morphological analysis in the form of identification, similarity, and clustering was conducted using MVSP and Minitab. Molecular analysis was conducted through PCR-ISSR amplification, followed by the calculation of PIC, RP, and genetic variation values. Dendrogram and PCoA were constructed using MVSP and GeneAlex. A total of 14 samples were found, with ten samples from the Progo river basin and four from the Opak river basin. The morphological characters that significantly contribute to the differences between samples are the branching pattern, presence of ventral stomata, ventral lobe color, ventral lobe length, and dorsal lobe shape. The species diversity is classified as low, with all samples identified as Azolla cristata. Among the ten Inter-Simple Sequence Repeats (ISSR) primers, UBC 814 and UBC 841 showed high PIC values (0.29; 0.28) and RP values (19.71; 23.29). All the primers used were effective in analyzing the genetic variation with an average polymorphism level of 85.27%. The unweighted pair group method analysis grouped the samples into two clusters based on their morphological and genetic traits. The genetic variation index is quite high in both populations, with Nei's gene diversity (He) = 0.230 and Shannon's information index (I) = 0.342. AMOVA revealed that 93% of the total genetic variation was found within populations, while only 7% was found between populations.

Kata Kunci : paku air, polimorfisme, variasi genetik, analisis klaster