ISOLASI GEN Aquaporin DARI Manihot esculenta Crantz cv UBI KUNING TERINDUKSI CEKAMAN KEKERINGAN

RISANG PANDEGAN, Dr. Endang Semiarti, MS., MSc

2014 | Skripsi | BIOLOGI

Komoditas Ubi Kayu yang bersifat resisten kekeringan sangat berpotensi untuk dikembangkan sebagai bahan baku utama industri pangan dan bioetanol, produksinya dapat ditingkatkan melalui bioteknologi ditengah tantangan krisis air dan perubahan iklim. Penelitian ini bertujuan mengisolasi gen aquaporin dari Manihot esculenta varietas Kuning serta mengkaji homologinya diantara sekuen-sekuen MeAQP. Cekaman kekeringan 26 hari mempengaruhi adaptasi fisiologis tanaman, terutama menurunkan laju pertambahan tinggi tanaman dan mempercepat laju gugur daun. Isolasi MeAQP diawali dengan isolasi mRNA MeAQP dari daun yang terinduksi cekaman kekeringan 26 hari, kemudian dilanjutkan first strand cDNA synthesis, optimasi kondisi PCR untuk RT-PCR dengan primer-primer gen spesifik, sekuensing gen dan diakhiri analisis homologi dengan Chromas, BioEdit dan CLUSTAL Omega. MeAQP berperan dalam menjaga tekanan turgor sel dan ketersediaan air. MeAQP cv Ubi Kuning yang toleran cekaman kekeringan bersifat homolog dengan gen aquaporin dari GeneBank, memiliki persentase similaritas antar sekuen gen yang tinggi (>70%) dan conserved domain berupa asam amino NPA (Asn-Pro-Aln)-amphipathic channel yang menunjukkan kemampuan resisten kekeringan.

Cassava is a drought resistance comodity, that is highly potential to be developed as raw material for food and bioethanol industries, Production of cassva can be enhanced through biotechnology within crisis of water and climate change challenges. The objectives of this research are to isolate aquaporin genes from Manihot esculenta cv Ubi Kuning here after called MeAQP gene, which is induced by drought stress and also to evaluate the homology among MeAQP sequences. 26 days of drought stress affects physiological adaptation, mainly to height development of plants and leaves fall rate. MeAQP isolation was initiated with MeAQP mRNA isolation from leaf that has been induced by dryness during 26 days, and then followed by first strand cDNA synthesis, optimizing PCR condition for RT-PCR with specific gene primers, gene sequencing and finally homology analysis using Chromas, BLAST, BioEdit and CLUSTAL Omega. MeAQP plays a role to keep cell turgidity pressure and water availability. MeAQP cv Ubi Kuning cell resistance to drought treatment is homolog with aquaporin gene from GeneBank, has high identity percentage among gene sequences (>70%) and has a conserved domain such as NPA amino acid (Asn-Pro-Aln)- amphipathic channel that indicate drought resistance capability.

Kata Kunci : Manihot esculenta, aquaporin, resisten kekeringan, gen.