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      Konstruksi padi nonaromatik yang beraroma wangi Menggunakan pcr berbantuan marka gen badh2

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      Date
      2009
      Author
      Hami Seno, Djarot Sasongko
      TJ, Santoso
      KR, TriJatmiko
      B, Padmadi
      D, Praptiwi
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      Abstract
      Tingginya permintaan pasar dan nilai komersial padi aromatik telah mendorong penelitian terkait aroma padi. Berbagai marka aromatik telah dilaporkan (Bradbury et al. 2005b, Lang and Buu 2008, Shi et al. 2008, Sakthivel et al. 2009). Namun selain aroma, sifat agronomi lain (produktivitas; waktu tanam; ketahanan hama dan penyakit; selektivitas area kultivasi, kemudahan tanam dan pemeliharaan; dsb.) padi aromatik tidak sebaik padi nonaromatik. Hal ini menjadi kendala bagi petani untuk menanam padi aromatik. Oleh karena itu pada penelitian ini dilakukan pengembangan varietas Ciherang aromatik/wangi dengan sifat agronomi sebaik host Ciherang ditambah sifat aroma dari donor Mentik Wangi. Konstruksi dilakukan melalui site-directed crossing berbantuan PCR dengan marker gen badh2, untuk menghindari produk transgenik. Seleksi dilakukan menggunakan PCR dengan primer spesifik gen badh2 (Bradbury et al. 2005). Pada tahun pertama (2009), DNA padi varietas nonaromatik (Ciherang) dan aromatik (Mentik Wangi, Gilirang, Pandan Wangi) diisolasi dari daun menggunakan metoda Shure et al. (1983), selanjutnya diamplifikasi dengan PCR menggunakan marka dan kondisi sesuai dengan yang dilaporkan oleh Bradbury et al. (2005). Padi aromatik yang dapat dibedakan dari padi nonaromatik kemudian disilangkan dengan padi nonaromatik Ciherang. Biji F1 kemudian ditanam, disolasi DNA nya, kemudian dianalisis dengan PCR menggunakan primer dan kondisi yang serupa. F1 yang heterozygot kemudian dibackcross dengan Ciherang, dan BC1F1 ditanam dan dianalisis PCR sebagaimana F1. Hasil analisis PCR mendapatkan bahwa hanya varietas aromatik Mentik Wangi yang dapat dibedakan dari varietas nonaromatik Ciherang, sedangkan Gilirang dan Pandan Wangi tidak. Selanjutnya Mentik Wangi digunakan sebagai donor aromatik. Analisis progeni F1 persilangan Ciherang-Mentik Wangi, mendapatkan heterozygot F1 yang memberikan pita amplifikasi dari Ciherang dan Mentik Wangi, sebagaimana dilaporkan oleh Bradbury et al. (2005b). Hasil serupa juga didapatkan pada analisis progeni BC1F1 Ciherang-Mentik Wangi. Pada tahun kedua (2010) dan ketiga (2011) akan diteruskan backcross hingga 4 x untuk mendapatkan BC5F1 dan diselfing untuk mendapatkan BC5F2 homozygot (Ciherang beraroma Mentik Wangi).
       
      The high commercial value and maket demmand of fragrant rice have triggered a number of researchs related to fragrant in rice. Various fragrant specific markers have been reported (Bradbury et al. 2005b, Lang and Buu 2008, Shi et al. 2008, Sakthivel et al. 2009). However, except for fragrant property, other agronomic traits (productivity; geographical location and period of cultivation; stress, insects, and disseases tolerance; simplcity of cultivation and maintenance, etc.) of non-fragrant rice are disadvantages compare to those of fragrant rice. These have become barriers for farmer to grow fragrant rice. Therefore, Ciherang rice possesing native agronomic traits and additional fragrant property, was developed in this research. To avoid transgenic plany product, construction was carried out through site-directed crosing and using badh2 specific marker-assisted PCR for progeny selection (Bradbury et al. 2005b). In the first year (2009), DNA of nonfragrant Ciherang and fragrant (Mentik Wangi, Gilirang, Pandan Wangi) rice were isolated from leaves and PCR amplified following the method as described by Shure et al. (1983) and Bradbury et al. (2005b), respectively. Fragrant rice with distinct PCR profile than that of non-fragrant Ciherang rice was then crossed to Ciherang. F1 seeds were planted, their DNA were isolated, and PCR analyzed as previous. The selected heterozygous F1 were then backcrossed to Ciherang, and the obtained BC1F1 were planted and analyzed as described for F1. PCR results showed that only Mentik Wangi pofile was disticnt compare to that of Ciherang, and therefore was selected as fragrant donor. In F1 progeny (Ciherang-Mentik Wangi) selection, heterozygot F1 with band amplification of Ciherang and Mentik Wangi, as described by Bradbury et al. (2005b), were obtained. The same results were obtained in BC1F1 (Ciherang-Mentik Wangi) progeny selection. In the 2nd (2010) and the 3rd (2011) year of research, BC1F1 will be further backcrossed 4 x and selfed, to obtain homozygous BC5F2 (Ciherang possesing Mentik Wangi fragrant).
       
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      http://repository.ipb.ac.id/handle/123456789/71535
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      Indonesia DSpace Group 
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