Ekspresi Gen Aquaporin pada Sengon (Falcataria Moluccana Miq) Resistan dan Terserang Hama Boktor Berbasis Real Time-PCR
Abstract
Sengon (F. moluccana) merupakan spesies tanaman cepat tumbuh (fast
growing) dan memiliki nilai ekonomi yang tinggi. Salah satu permasalahan yang
terjadi pada penanaman sengon monokultur adalah timbulnya serangan hama
boktor (Xystrocera festiva). Secara alami tanaman sengon memiliki gen ketahanan
terhadap serangan hama boktor, salah satunya adalah aquaporin. Tujuan penelitian
ini adalah mengukur tingkat ekspresi gen aquaporin pada kambium pohon sengon
resistan dan terserang hama boktor secara kuantitatif dengan teknik real-time PCR.
Prosedur penelitian ini meliputi isolasi RNA total, sintesis cDNA, perancangan
primer spesifik gen aquaporin dan aktin, optimasi eskpresi gen dengan real-time
PCR, dan dianalisis menggunakan metode Comparative CT method (2
-ΔΔC
T). Hasil
kuantifikasi menunjukkan tingkat ekspresi gen aquaporin pada ketiga sampel
sengon resistan yaitu sebesar 573,05; 2,62; dan 177,99 sedangkan sengon terserang
boktor sebesar 8,04; 3,76; dan 4,67. Secara garis besar, ekspresi gen aquaporin pada
sengon resistan lebih tinggi dibandingkan sengon terserang hama boktor, kecuali
pada sampel 2. Sengon (F. moluccana) is a fast-growing plant species with high economic
value. One problem in monoculture sengon planting is the emergence of boktor
pests (Xystrocera festiva). Naturally, sengon plants have genes for resistance to
boktor pests, one of which is aquaporin. This study aimed to measure the level of
aquaporin gene expression in the cambium of resistant and attacked boktor pests
quantitatively using the Real-time PCR technique. The research procedures
included total RNA isolation, cDNA synthesis, design of specific primers for
aquaporin and actin genes, optimization of gene expression with real-time PCR,
and analysis using the Comparative CT method (2
-ΔΔC
T). The quantification results
showed the expression levels of the aquaporin gene in the three samples of resistant
sengon were 573,05; 2,62; and 177,99, while sengon attacked by boktor were 8,04;
3,76; and 4,67. In general, expression of the aquaporin gene in resistant sengon was
higher than that of sengon attacked by boktor, except for sample 2.
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