Genetic Diversity of Artemisia annua L. and Artemisia vulgaris L. Based on Amplified Fragment Length Polymorphism (AFLP) and Morphological Traits

Abstract

Malaria remains a serious problem in Indonesia. Artemisinin is an antimalarial compound that is able to treat malaria disease. Until now, Artemisia annua is still the only one source of artemisinin. On the other hand Artemisia vulgaris is Artemisia species that grows widely in Indonesia. The introduction of A. annua from China to Indonesia produces diverse phenotypes and unstable artemisinin content. The objective of this research was to analyse the genetic diversity of A. annua and A. vulgaris based on Amplified Fragment Length Polymorphism (AFLP) and morphological traits. Amplification was performed by using the labeled P11 primer IRD 700 and three selective primers, M-CAC, M-CAG and M-CAT. Three morphological traits were observed. The data from AFLP and morphological traits were translated into binary data. Similarity matrix analysis was carried out by using the software NTSYSpc version 2.02i. Principal Component Analysis was done by using the Minitab 14 program. By using these three selective primers, 111 AFLP loci were amplified. These loci can not clustered Artemisia into its species. There is not any specific loci addressed to specific accession from 111 AFLP loci, but from 48 AFLP loci, locus number 38 can be used as specific marker for three accessions consist of A. vulgaris accession which has light brown stems and wide leaves (VCOL), A. vulgaris accession which has light brown stems and narrow leaves (VCOS) and A. annua accession which has purple green stems and narrow leaves (AHUS). Analysis of 63 AFLP loci consist of loci number 49-111, showed that loci number 101, 103 and 109 addressed to A. annua accession which has purple green stems and wide leaves that arranged like a roset (AHULr). Based on morphological traits, the diversity between A. annua and A. vulgaris was 39%, while the diversity within A. annua species was 29%. Analysis of morphological traits and 48 AFLP loci showed that the A. annua accession which has purple green stems and wide leaves that arranged like a roset (AHULr) not cluster into A. annua species or A. vulgaris species.

Penyakit malaria masih merupakan masalah serius di Indonesia. Artemisinin merupakan senyawa antimalaria yang mampu mengobati penyakit malaria. Sampai saat ini Artemisia annua masih merupakan satu-satunya sumber artemisinin. A. annua adalah tanaman hari pendek yang berasal dari China. Introduksi A. annua ke daerah tropik menyebabkan tanaman cepat berbunga sehingga kandungan artemisinin turun. Di Indonesia ada lima aksesi A. annua hasil introduksi yang dikoleksi oleh Balai Besar Penelitian dan Pengembangan Tanaman Obat dan Obat Tradisional (BBPPTO-OT) Tawangmangu, Solo. Aksesi ini bervariasi pada warna batang, ukuran relatif daun dan susunan daun pada batang. Selain itu terdapat perbedaan kerapatan trikoma kelenjar dan kandungan artemisinin antara aksesi hijau dan aksesi ungu. Belum ada data mengenai keragaman genetik antar aksesi A. annua hasil introduksi di Indonesia. Artemisia vulgaris adalah jenis Artemisia yang ada di Indonesia yang dikenal dengan nama daerahnya sudamala. Herba ini tersebar hampir di semua dataran tinggi di Indonesia namun paling banyak ditemukan di Papua. BBPPTO-OT Tawangmangu, Solo, memiliki dua aksesi A. vulgaris, yaitu aksesi berdaun lebar dan aksesi berdaun sempit. Kandungan artemisinin A. vulgaris jauh lebih rendah dibandingkan dengan A. annua, namun spesies ini memiliki potensi sebagai sumber artemisinin lokal karena tumbuh secara alami di Indonesia. Belum ada laporan mengenai hubungan kekerabatan A. annua hasil introduksi di Indonesia dengan A. vulgaris.