Use of microsatellite markers to detect heterozygosity in an F2 generation of a black rice and white rice cross

https://doi.org/10.22146/ijbiotech.33111

Kristamtini Kristamtini(1*), Taryono Taryono(2), Panjisakti Basunanda(3), Rudi Hari Murti(4)

(1) Assessment Institute for Agricultural Technology Yogyakarta, Jalan Stadion Maguwoharjo No. 22, Wedomartani, Ngemplak, Sleman, Yogyakarta
(2) Agronomy Department, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora, Bulaksumur, Yogyakarta 55281
(3) Agronomy Department, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora, Bulaksumur, Yogyakarta 55281
(4) Agronomy Department, Faculty of Agriculture, Universitas Gadjah Mada, Jalan Flora, Bulaksumur, Yogyakarta 55281
(*) Corresponding Author

Abstract


The aim of this research was to know the heterozygosity of F2  generation from black rice and white rice crossing using microsatellite marker.  The research material consisted of F2 Sx G plant population from black rice (S) and white rice Situbagendit (G) crosses, female parent of black rice (S), male parent of white rice (G), chemical and organic fertilizer, chemicals and tools for molecular activity and 3 microsatellite markers related to color properties  (RM 220, RM 224 and RM 252). All of plant populations (generation F2, parent female, parent male) were planted in fields up to harvest. Young leaves (30 days after planting) all of plant populations were molecularly analyzed using 3 microsatellite markers (RM 220, RM 224 and RM 252). Stages of this activity include DNA isolation, PCR reaction, and visualization of PCR results using Metaphore Agarose Gel Electrophoresis. The results showed that the percentage of the number of individual plants showing heterozygous pattern in F2 S × G plant generation was 50% (RM 220); 40% (RM 224) and 60% (RM 252), so the RM 252 microsatellite marker was effectively used as a DNA-assisted selection tool on the crossbreed of black rice with white rice.


Keywords


black rice; F2 generation; heterozygosity; microsatellite

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Kristamtini et al.


References

Allard Robert W., Allard Robert Wayne. 1999. Principles of plant breeding. John Wiley & Sons.

Azrai M. 2006. Sinergi teknologi marka molekuler dalam pemuliaan tanaman jagung [Sinergy of molecular marker technology in corn breeding in Indonesia]. Jurnal Litbang Pertanian 25:81--89.

Doyle J. 1990. Isolation of plant DNA from fresh tissue. Focus 12:13–15.

Gomez K, Gomez A. 2007. Statistical procedures for agricultural research [Prosedur statistik untuk penelitian pertanian]. 2nd ed. Sjamsuddin E, Baharsjah J, translators. Jakarta: UI Press.

Ichikawa H, Ichiyanagi T, Xu B, Yoshii Y, Nakajima M, Konishi T. 2001. Antioxidant activity of anthocyanin extract from purple black rice. Journal of Medicinal Food 4:211–218. doi:10.1089/10966200152744481.

Kristamtini, Purwaningsih H. 2009. Potensi pengembangan beras merah sebagai plasma nutfah Yogyakarta [Potential of red rice development as Yogyakarta’s germplasm]. J Litbang Pertanian 28:88--95.

Lapitan VC, Brar DS, Abe T, Redoña ED. 2007. Assessment of genetic diversity of Philippine rice cultivars carrying good quality traits using SSR markers. Breeding Science. 57:263–270. doi:10.1270/jsbbs.57.263.

McCouch SR. 2002. Development and Mapping of 2240 New SSR markers for rice (Oryza sativa L.). DNA Research 9:199–207. doi:10.1093/dnares/9.6.199.

Ryu SN, Park SZ, HO C-T. 1998. High performance liquid chromatographic determination of anthocyanin pigments in some varieties of black rice. Journal of food and Drug Analysis 6(4): 729–736.

Susanto U, Sutrisno, Aswidinnoor H. 2009. Pemanfaatan teknik markah molekuler untuk perbaikan varietas padi [Utilization of molecular marking technology for the improvement of rice varieties]. Subang: Publikasi Balai Besar Penelitian Tanaman Padi.

Suzuki M, Kimura T, Yamagishi K, Shinmoto H, Yamaki K. 2004. Comparison of mineral contents in 8 cultivars of pigmented brown rice. Nippon Shokuhin Kagaku Kogaku Kaishi 51:424–427. doi:10.3136/nskkk.51.424.

Tuinstra MR, Grote EM, Goldsbrough PB, Ejeta G. 1996. Identification of quantitative trait loci associated with pre-flowering drought tolerance in sorghum. Crop Science 36:1337. doi:10.2135/cropsci1996.0011183X003600050043x.

Utami DW, Kristamtini, Prajitno. 2009. Karakterisasi plasma nutfah padi beras merah lokal asal propinsi daerah istimewa yogyakarta berdasarkan karakter morfo-agronomi dan marka SSRs [Characterizing local red rice germplasm from Yogyakarta Special Province based on morpho-agronomical characteristics and SSRs marker]. Zuriat 20:10--18.

Wiriyasuk K. 2005. Regulation of genes controlling grain anthocyanin and proanthocyanidin (condensed tannins) accumulation in rice. [Kasetsart]: Kasetsart University.

Zhou H, Xie Z, Ge S. 2003. Microsatellite analysis of genetic diversity and population genetic structure of a wild rice (Oryza rufipogon Griff.) in China. Theoretical and Applied Genetics 107:332–339. doi:10.1007/s00122-003-1251-y.



DOI: https://doi.org/10.22146/ijbiotech.33111

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