Pengaruh Metil Jasmonat Terhadap Pertumbuhan dan Ekspresi Gen PAL Pada Kultur In Vitro Daun Dewa (Gynura pseudochina (L.) DC)

Salma, Argya Rasmi (2023) Pengaruh Metil Jasmonat Terhadap Pertumbuhan dan Ekspresi Gen PAL Pada Kultur In Vitro Daun Dewa (Gynura pseudochina (L.) DC). Other thesis, Institut Teknologi Sepuluh Nopember.

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Abstract

Pemakaian tanaman obat sebagai bahan baku pengobatan tradisional semakin digemari. Gynura pseudochina (L) DC. dengan nama lokal daun dewa adalah salah satu tanaman obat tradisional yang terkenal di Asia Tenggara yang mengandung komponen senyawa bioaktif metabolit sekunder seperti flavonoid, alkaloid, saponin, dan tanin. Berbagai teknik dan metode dikembangkan untuk mendapatkan bibit tanaman yang bebas hama penyakit guna meningkatkan mutu fitomedisin, salah satunya dengan kultur in vitro. Sementara itu diperlukan elisitor untuk meningkatkan kuantitas metabolit sekunder G. pseudochina. Salah satu elisitor yang dapat digunakan adalah metil jasmonat (MeJA). MeJA dilaporkan dapat mempengaruhi pembentukan organ serta meningkatkan ekspresi gen PAL pada tumbuhan. Aktivitas PAL diketahui akan meningkat setelah tanaman diberikan MeJA dan berdampak pada produksi berlebih senyawa flavonoid. Penelitian ini bertujuan untuk mengetahui pengaruh MeJA terhadap pertumbuhan dan ekspresi gen PAL G. pseudochina secara in vitro. Penelitian ini dilakukan dengan pembuatan media Murashige dan Skoog (MS) yang ditambahkan IAA 0,25 ppm dan BAP 3 ppm serta MeJA dengan konsentrasi yang berbeda yaitu 0, 75, 150 dan 300 µM, pengukuran parameter pertumbuhan berupa berat basah, tinggi tanaman, jumlah daun, akar dan tunas, serta persentase kalogenesis dan organogenesis eksplan. Kemudian dilakukan pengamatan ekspresi gen dengan melakukan ekstraksi RNA total dan Real-Time PCR kuantitatif (qRT-PCR). Data dianalisis secara kualitatif berupa data ekspresi gen dan kuantitatif berupa data parameter pertumbuhan dengan menggunakan ANOVA one way yang dilanjutkan dengan post-hoc Tukey test. Parameter ekspresi gen didapat berdasarkan data hasil qRT-PCR yang sebelumnya telah dihitung menggunakan metode 2 -ΔΔCT, yang dilakukan oleh Livak & Schmittgen (2001). Hasil yang didapat menunjukkan pemberian metil jasmonat pada G. pseudochina secara in vitro berpengaruh pada kalogenesis dan organogenesis. Pertumbuhan optimal kalogenesis adalah pemberian MeJA 150 µM berdasarkan persentase tumbuh kalus, tekstur kalus, dan warna kalus. Sedangkan pada organogenesis, persentase pertumbuhan akar tertinggi yaitu konsentrasi 75 µM dan persentase pertumbuhan tunas tertinggi adalah konsentrasi 150 µM. Pada parameter pertumbuhan, diketahui MeJA memberikan pengaruh signifikan (P<0,05). Konsentrasi MeJA 150 µM memberikan pengaruh yang optimal terhadap parameter tinggi tanaman, berat basah, jumlah daun, dan jumlah tunas serta konsentrasi MeJA 75 µM memberikan pertumbuhan akar yang optimal. Ekspresi gen PAL pada tanaman G. pseudochina pasca pemberian MeJA mengalami peningkatan ekspresi gen relatif. Ekspresi gen tertinggi adalah MeJA konsentrasi 150 µM dengan peningkatan 1,29 fold dibandingkan dengan perlakuan kontrol (MeJA 0 µM).
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The use of medicinal plants as raw materials for traditional medicine is currently become increasingly popular. Gynura pseudochina (L) DC. with the local name daun dewa is one of the well-known traditional medicinal plants in Southeast Asia which contains components of secondary metabolites such as flavonoids, alkaloids, saponins and tannins. Various techniques and methods have been developed to obtain pest-free plant to improve the quality of phytomedicins, one of which is by in vitro culture. Meanwhile, an elicitor is needed to increase the quantity of secondary metabolites G. pseudochina. One of the elicitors that can be used is methyl jasmonate (MeJA). MeJA is reported to affect organ formation and increase PAL gene expression in plants. PAL activity is known to increase after the plants are given MeJA and have an impact on the excess production of flavonoid compounds. This study aims to determine the effect of MeJA on the growth and expression of the PAL gene of G. pseudochina in vitro. This research was conducted by preparing Murashige and Skoog (MS) media added with IAA 0,25 ppm and BAP 3 ppm and MeJA with different concentrations of 0, 75, 150 and 300 µM, measuring growth parameters in the form of fresh weight, plant height, number of leaves, roots and shoots, percentage of callogenesis and organogenesis. Then the gene expression was observed by performing total RNA extraction and quantitative Real-Time PCR (qRT-PCR). The data were analyzed qualitatively in the form of gene expression data and quantitatively in the form of growth parameter data using one way ANOVA followed by a post-hoc Tukey test and the parameters of gene expression were obtained based on qRT-PCR results which had previously been calculated using the method 2 -ΔΔCT, conducted by Livak & Schmittgen (2001). The results showed that methyl jasmonate giving into G. pseudochina in vitro had an effect on callogenesis and organogenesis. Optimal growth of callogenesis was MeJA 150 µM based on the percentage of callus growth, callus texture, and callus color. Whereas in organogenesis, the highest percentage of root growth was concentration of 75 µM MeJA and the highest percentage of shoot growth was concentration of 150 µM MeJA. On the growth parameters, it is known that MeJA has a significant effect (P<0.05). The MeJA concentration of 150 µM has an optimal effect on the parameters of plant height, fresh weight, number of leaves, and number of shoots and the MeJA concentration of 75 µM provides optimal root growth. PAL gene expression in G. pseudochina under treatment of MeJA was found increased. The highest gene expression was MeJA at concentration of 150 µM with an increase of 1.29 fold compared to the control treatment (MeJA 0 µM).

Item Type:	Thesis (Other)
Uncontrolled Keywords:	Gynura pseudochina, Ekspresi Gen PAL, Kultur In Vitro, Metil Jasmonat, Pertumbuhan; Growth, In Vitro Culture, Methyl Jasmonate, PAL Gene Expression
Subjects:	Q Science > QH Biology S Agriculture > SB Plant culture
Divisions:	Faculty of Science and Data Analytics (SCIENTICS) > Biology > 46201-(S1) Undergraduate Thesis
Depositing User:	SALMA ARGYA RASMI
Date Deposited:	04 Oct 2023 02:12
Last Modified:	04 Oct 2023 02:12
URI:	http://repository.its.ac.id/id/eprint/102542

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