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"Penelitian terhadap Hibiscus rosa-sinensis variasi crested peach pada musim kemarau dan hujan menemukan plastisitas fenotipe, kemampuan genotipe untuk menunjukkan fenotipe yang berbeda berdasarkan lingkungannya. Plastisitas fenotipe tersebut menyebabkan karakter bunga menyerupai bunga single atau double dilihat dari perubahan stamen menjadi petal serta morfologi dan anatomi ovarium. Perubahan tersebut diduga disebabkan oleh perubahan ekspresi gen homeotik MADS-box, tepatnya gen AGAMOUS. Oleh karena itu, dilakukan penelitian untuk menganalisis pengaruh lingkungan terhadap plastisitas fenotipe serta hubungan antara plastisitas fenotipe dan ekspresi gen AGAMOUS pada bunga tersebut. Pencatatan suhu udara dan tanah, kelembapan udara dan tanah, pH tanah, dan intensitas cahaya serta morfologi bunga dan ovarium dilakukan setiap hari Senin, Rabu, dan Jumat. Isolasi DNA dan PCR dilakukan untuk menemukan primer optimal dalam amplifikasi gen AGAMOUS. Isolasi RNA dan RT-PCR dilakukan untuk menemukan primer yang menempel pada ekson. Hasil penelitian menunjukkan kombinasi suhu udara, kelembapan udara, dan curah hujan mempengaruhi plastisitas fenotipe bunga, berdasarkan data morfologi dan data ovarium. Amplifikasi DNA dengan primer A1, C6, dan C7 masing-masing menghasilkan satu pita dengan panjang 300 bp, 200 bp, dan 300 bp. Primer D3 menghasilkan pola pita yang unik; bunga single memiliki pita dengan panjang 550 bp dan 450 bp, sedangkan crested dan double memiliki pita tambahan dengan panjang 300 bp. Amplifikasi cDNA dengan sembilan primer yang diuji tidak menghasilkan pita yang terdeteksi. Pola pita primer D3 diduga merupakan gene duplication atau copy number variation pada gen AGAMOUS yang mungkin belum diekspresikan tetapi ada sebagai salah satu cara tumbuhan bertahan hidup di tengah perubahan iklim.

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Research on Hibiscus rosa-sinensis crested peach found that changes of stamen into additional petal and ovary morphology and anatomy cause flowers to resemble single or double flowers, depending on the season. Such organ changes are assumed to be caused by MADS-box homeotic gene expression changes, specifically the AGAMOUS gene. Therefore, research was done to analyze the effects of the environment on phenotypic plasticity and analyze the relationship between phenotypic plasticity and AGAMOUS gene expression. The study was conducted by observing and linking air and soil temperature, air and soil humidity, soil pH, and light intensity with flower and ovary morphology every Monday, Wednesday, and Friday. DNA isolation and PCR were conducted to find optimal primers for amplifying AGAMOUS gene. RNA isolation and RT-PCR were conducted to find primers that attach to exons. Results found that a combination of air temperature, air humidity, and rainfall affects the phenotypic plasticity, as seen from the variations of flower and ovary morphology that occurred. DNA amplification with primers A1, C6, and C7 produced one band with a length of 300 bp, 200 bp, and 300 bp, respectively. Primer D3 produced a unique banding pattern; single flowers have bands with lengths of 550 bp and 450 bp, while crested and double flowers have an additional band with a length of 300 bp. cDNA amplification with nine primers tested did not produce detectable bands. The difference in the banding pattern produced by primer D3 is suspected to be a gene duplication or copy number variation in the AGAMOUS gene that may not have been expressed but exists as one way for plants to survive amidst climate change."

"Bunga Hibiscus rosa-sinensis L. variasi crested peach dan double orange berbeda dari variasi single pink karena memiliki petal tambahan petaloid . Struktur petaloid tersebut diduga berasal dari modifikasi organ reproduktif bunga homeosis . Peristiwa homeosis yang terjadi dihipotesiskan karena gen kelas C AGAMOUS yang berperan dalam pembentukan androecium dan gynoecium tidak terekspresi. Oleh karena itu dilakukan penelitian yang bertujuan untuk mengetahui ekspresi gen AGAMOUS secara kualitatif pada bunga single pink, crested peach, dan double orange. Analisis ekspresi gen AGAMOUS dilakukan dengan cara mengisolasi RNA dari androecium dan gynoecium ketiga variasi bunga menggunakan metode CTAB yang dimodifikasi. Sampel RNA diubah menjadi cDNA menggunakan reverse transcriptase, yang selanjutnya diamplifikasi dengan teknik PCR menggunakan primer AG1 dan AG2. Produk PCR AG1 menghasilkan variasi pita dengan ukuran 100, 200, dan 300 bp, sedangkan hasil PCR AG2 menghasilkan pita yang berukuran 200 bp. Hasil analisis sekuensing terhadap produk PCR primer AG1 dan AG2 menunjukkan gen AGAMOUS terekspresi pada semua sampel. Hasil tersebut mengindikasikan bahwa perubahan struktur organ reproduktif bunga tidak disebabkan oleh hilangnya ekspresi gen AGAMOUS, sehingga perlu dilakukan analisis ekspresi gen AGAMOUS beserta interaksinya dengan gen lain.

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Hibiscus rosa sinensis L. crested peach and double orange types are different from single pink type in terms of their additional petals petaloid . The petaloid structure is thought to have originated from reproductive organs modification homeosis . AGAMOUS is class C gene that plays role in androecium and gynoecium formation. Loss of AGAMOUS gene expression is assumed to cause modifications occur in reproductive organs. Therefore, this study aims to determine the qualitative expression of AGAMOUS gene on single pink, crested peach, and double orange flowers. Analysis of AGAMOUS gene expression was done by isolating RNA from their androecium and gynoecium using the modified CTAB method. The RNA sample was converted to cDNA using reverse transcriptase, before further amplified by PCR technique using AG1 and AG2 primers. The AG1 PCR product produces bands of 100, 200, and 300 bp, while the PCR AG2 produces single band of 200 bp. The analysis of sequencing results showed that AGAMOUS gene expressed in all samples. Therefore, petaloids presents in crested peach and double orange flowers are not caused by loss of AGAMOUS gene expression. The homeosis occurred should be analyzed not only based on AGAMOUS gene expression, but also should include other gene and their interactions. "

"Penelitian dilakukan untuk mengetahui variasi bunga Hibiscus rosa-sinensis L. bentuk crested di alam, serta hubungannya dengan ekspresi gen MADS-box. Sebanyak 200 sampel bunga crested diamati secara morfologi melalui pengukuran panjang staminal column, perhitungan perhiasan bunga, dan pengamatan visual permukaan luar ovarium. Sebanyak 137 sampel bunga crested diamati secara anatomi melalui pengamatan visual struktur internal ovarium. Hasil pengamatan morfologi dan anatomi menunjukkan tingginya variasi bunga H. rosa-sinensis bentuk crested. Salah satu karakter yang menentukan tingginya variasi bunga crested di alam adalah keberadaan petal tambahan yang diduga tumbuh di lokasi keberadaan stamen dan petal akibat gejala homeosis. Homeosis pada bunga crested diduga karena tidak terekspresikannya gen C yang merupakan salah satu kelas dari gen MADS-box. Untuk membuktikan dugaan tersebut, dilakukan pengamatan molekular melalui analisis gen MADS-box yang berperan dalam proses pembungaan. Dari kelima kelas gen MADS-box yang diamati, hanya gen C yang berhasil diamplifikasi. Hasil menunjukkan bahwa gen C terekspresi di semua bagian bunga crested. Berdasarkan hasil tersebut, homeosis pada bunga crested bukan disebabkan karena tidak terekpresikannya gen C. Dibutuhkan penelitian lebih lanjut untuk mengetahui tingkat ekspresi tiap kelas gen MADS-box terhadap variasi bunga crested di alam.

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The aim of this study is to know the variation of Hibiscus rosa-sinensis L. crested flower in nature, and to know its correlation against MADS-box gene expression. The study was conducted through morphological, anatomical, and molecular observation. Morphological sections were carried out on 200 samples of flowers by measuring the length of staminal column, counting the number of perianthium, and observing the external structure of ovaries. Anatomical sections were carried out on 137 samples of flowers by observing the internal structure of ovaries. The results showed that H. rosa-sinensis crested flower has a high variety in the shape of flower. Two main parts of crested flower that effecting its variety in nature are staminodium petaloid and stamen-petal intermediate that lied in the position of stamen and petal. This phenomenon is assumed as homeotic due to the absence of C gene expression that belong to MADS-box gene family. Based on molecular observation, AGAMOUS gene (MADS-box class C) expressed in all of crested flower parts, including staminodium petaloid and stamen-petal intermediate. This results is on contradiction with the assumption that homeosis in H. rosa-sinensis caused by the absence of gene C. Further research is needed to know the expression of others MADS-box class genes expression, including their level of expression in each parts of crested flower."