Sains Malaysiana 51(12)(2022): 3937-3948

http://doi.org/10.17576/jsm-2022-5112-06

Doubled Haploid Rice Lines Production through Anther Culture of F₁ Derived from Abiotic Stress Tolerant Parents

(Pengeluaran Titis Beras Haploid Berganda melalui Kultur Anter Terbitan daripada Induk Tahan Tekanan Abiotik)

CUCU GUNARSIH^1,2, BAMBANG SAPTA PURWOKO^1,*, ISWARI SARASWATI DEWI³, WILLY BAYUARDI SUWARNO¹ & NAFISAH²

¹Department of Agronomy and Horticulture, Faculty of Agriculture, Bogor Agricultural University, Kampus IPB Darmaga, Bogor 16680, Indonesia

²Indonesian Center for Rice Research, Sukamandi, Jl. Raya IX, Sukamandi Subang, Indonesia

³Research Center for Food Crops, National Research and Innovation Agency (BRIN), Cibinong Science Center – Botanical Garden, Jl. Raya Jakarta-Bogor No. Km. 46, Cibinong, Bogor 16911, Indonesia

Diserahkan: 28 Oktober 2021/Diterima: 19 Ogos 2022

Abstract

Anther culture is one of the feasible technologies for accelerating rice breeding programs. This research aimed at determining the anther culture ability and doubled-haploid plants production of eight F₁s derived from single crosses between indica rice parents that had a single or multiple abiotic stresses tolerance. The experiment was conducted using completely randomized design (CRD) with 15 replications. Medium for callus induction was based on N6 + 2.0 mg L^-1 NAA +0.5 mg L^-1 kinetin + 1 mM putrescine, while the regeneration medium was based on MS + 0.5 mg L^-1 NAA + 2.0 mg L^- kinetin ¹ + 1 mM putrescine. Medium for rooting was MS + 0.5 mg L^-1 IBA. The results indicated that all F₁ populations gave varied responses to callus induction and plant regeneration. The F₁populations from HS4-11-1-2/B13926E-KA-23 had higher anther culture ability compared to other seven F₁ populations as indicated by the efficiency in callus formation and plant regeneration. Overall, the number of doubled haploid individuals (DHs) obtained was 158. F₁ populations using HS4-11-1-2 as female parent, HS4-11-1-2/B13926E-KA-23 and HS4-11-1-2/CG8-93-1-1 contributed 45.4% and 34.9% doubled haploid individuals, respectively. The developed DHs lines need to be morphologically and agronomically characterized as well as screened under different combinations of abiotic stress to identify high-yielding and abiotic stress tolerant individuals for further rice breeding program.

Keywords: Anther culture ability; callus induction; characterization; doubled haploid; rice breeding

Abstrak

Kultur anter adalah salah satu teknologi yang boleh dilaksanakan untuk mempercepatkan program pembiakbakaan padi. Kajian ini bertujuan untuk menentukan kemampuan kultur anter dan pengeluaran tanaman haploid berganda bagi lapan populasi F₁ yang berasal daripada persilangantunggal induk padi indica yang tahan terhadap tekanan abiotik tunggal atau berganda. Uji kaji ini dijalankan menggunakan reka bentuk rawak lengkap (CRD) dengan 15 replikasi. Medium untuk aruhan kalus adalah berdasarkan N6 + 2.0 mg L^-1 NAA+ 0.5 mg L^-1 kinetin+ 1 mM putrescine, manakala medium penjanaan semula adalah berasaskan MS + 0.5 mg L^-1 NAA + 2.0 mgL^-1 kinetin + 1 mM putrescine. Medium untuk pengakaran ialah MS + 0.5 mg L^-1 IBA. Keputusan menunjukkan bahawa semua populasi F₁ memberikan tindak balas yang berbeza-beza terhadap aruhan kalus dan penjanaan semula tumbuhan. Populasi F₁ daripada HS4-11-1-2/B13926E-KA-23 mempunyai keupayaan kultur anter yang lebih tinggi berbanding tujuh populasi F₁ lain seperti yang ditunjukkan oleh kecekapan dalam pembentukan kalus dan penjanaan semula tumbuhan. Secara keseluruhan, bilangan individu haploid berganda (DH) yang diperoleh adalah sebanyak 158. Populasi F₁ yang menggunakan HS4-11-1-2 sebagai induk betina, HS4-11-1-2/B13926E-KA-23 dan HS4-11-1-2/CG8-93-1-1 masing-masing menyumbang 45.4% dan 34.9% individu DH. Titisan DH yang dibangunkan perlu dicirikan secara morfologi dan agronomi serta disaring di bawah gabungan tekanan abiotik yang berbeza untuk mengenal pasti individu berhasil tinggi dan tahan terhadap tekanan abiotik untuk program pembiakbakaan padi selanjutnya.

Kata kunci: Aruhan kalus; haploid berganda; kemampuan kultur anter; pembiakbakaan padi

RUJUKAN

Akbar, M.R., Purwoko, B.S., Dewi, I.S., Suwarno, W.B. & Sugiyanta. 2018. Agronomic and drought tolerance evaluation of doubled haploid rice breeding lines derived from anther culture. SABRAO Journal of Breeding and Genetics 50(2): 115-128.

Ali, J., Nicolas, K.L.C., Akther, S., Torabi, A., Ebadi, A.A., Marfori-Nazarea, C.M. & Mahender, A. 2020. Improved anther culture media for enhanced callus formation and plant regeneration in rice (Oryza sativa L.). Plant 10(5): 839.

Ali, J., Xu, J.L., Gao, Y.M., Ma, X.F., Meng, L.J., Wang, Y., Pang, Y.L., Guan, Y.S., Xu, M.R., Revilleza, J.E., Franje, N.J., Zhou, S.C. & Li, Z.K. 2017. Harnessing the hidden genetic diversity for improving multiple abiotic stress tolerance in rice (Oryza sativa L.). PLoS ONE 12(3): e0172515.

Anshori, M.F., Purwoko, B.S., Dewi, I.S., Ardie, S.W. & Suwarno, W.B. 2021. A new approach to select doubled haploid rice lines under salinity stress using indirect selection index. Rice Science 28(4): 368-378.

Anshori, M.F., Purwoko, B.S., Dewi, I.S., Suwarno, W.B. & Ardie, S.W. 2019. Cluster heatmap for detection of good tolerance trait on doubled-haploid rice lines under hydroponic salinity screening. IOP Conf. Series: Earth and Environmental Science 484: 1-7.

Anshori, M.F., Purwoko, B.S., Dewi, I.S., Ardie, S.W., Suwarno, W.B. & Safitri, H. 2018. Determination of selection criteria for screening of rice genotypes for salinity tolerance. SABRAO Journal of Breeding and Genetics 50(3): 279-294.

BBPADI. 2018. Varietas inbrida padi sawah irigasi. http://bbpadi.litbang.pertanian.go.id/index.php/varietas/inbrida-padi-sawah-irigasi-inpari/content/item/513-inpari-42-agritan-gsr). Accessed on 13 September 2018.

BPS. 2020. Berita Resmi Statistik. BPS. Jakata. from https://www.bps.go.id/brs.html. Accessed on 28 January 2021.

Chaikam, V., Molenaar, W., Melchinger, A.E.  & Boddupalli, P.M. 2019. Doubled haploid technology for line development in maize: technical advances and prospects. Theoretical and Applied Genetics 132(12): 3227-3243.

Collard, B.C.Y., Beredo, J.C., Lenaerts, B., Mendoza, R., Santelices, R., Lopena, V., Verdeprado, H., Raghavan, C., Gregorio, G.B., Vial, L., Demont, M., Biswas, P.S., Iftekharuddaula, K.M., Rahman, M.A., Cobb, J.N. & Islam, M.R. 2017. Revisiting rice breeding methods: Evaluating the use of rapid generation advance (RGA) for routine rice breeding. Plant Production Science 20(4): 337-352.

Dewi, I.S. & Purwoko, B.S. 2012. Kultur antera untuk percepatan perakitan varietas padi di Indonesia. Jurnal Agro Biogen 8(2): 78-88.

Dewi, I.S. & Purwoko, B.S. 2011. Kultur in vitro untuk produksi tanaman haploid androgenic. In Bioteknologi dalam Pemuliaan Tanaman, edited by Wattimena, G.A., Nurhajati, A.M., Wiendi, N.M.A., Purwito, A., Efendi, D., Purwoko, B.S. & Khumaida, N. IPB Press. Bogor Indonesia. pp. 107-157.

Dewi, I.S. & Purwoko, B.S. 2008. Role of polyamines in inhibition of ethylene biosynsthesis and their effects on rice anther culture development. Indonesian Journal of Agricultural Science 9(2): 60-67.

Dewi, I.S., Safitri, H. & Purwoko, B.S. 2020. Effect of sucrose on calus induction and green plantlet regeneration in anther culture of Indica × Indica rice. IOP Conf. Ser.: Earth Environ. Sci. 484: 012023.

Dewi, I.S., Putri, N.H. & Purwoko, B.S. 2019. Response of anther donor genotypes (F₁) from Indica × Indica crosses to rice anther culture. Jurnal AgroBiogen 15(1): 45-52.

Dewi, I.S., Purwoko, B.S. & Bakhtiar. 2006. Development of aluminium tolerant upland rice through anther culture. In Proc. International Rice Conference. Bali-Indonesia, 12-14 September 2006.

Dewi, I.S., Syafii, M., Purwoko, B.S. & Suwarno, W.B. 2017. Efficient indica rice anther culture derived from three-way crosses. SABRAO Jurnal of Breeding and Genetics 49(4): 336-345.

Dewi, I.S., Purwoko, B.S., Aswidinoor, H. & Somantri, I.H. 2004. Kultur antera padi pada beberapa formulasi media yang mengandung poliamin. Jurnal Bioteknologi Pertanian 9(1): 14-19.

Germana, M.A. 2011. Anther culture for haploid and doubled haploid production. Plant Cell Tissue Organ Culture (PCTOC) 104: 283-300.

Gunarsih, C., Purwoko, B.S., Dewi, I.S. & Syukur, M. 2016. Regenerasi dan aklimatisasi kultur antera enam persilangan F₁ padi sawah. Jurnal Agronomi Indonesia 44(2): 133-140.

Gueye, T. & Ndir, K.N. 2010. In vitro production of double haploid plants from two rice species (Oryza sativa L. and Oryza glaberrima Steudt.) for the rapid development of new breeding material. Scientific Research and Essays 5: 709-713.

Grewal, D., Manito, C. & Bartolome, V. 2011. Doubled haploids generated through anther culture from crosses of elite indica and japonica cultivars and or lines of rice: Large-scale production, agronomic performance, and molecular characterization. Crop Science 51: 2544-2553.

Ilham, N., Syaukat, Y. & Friyatno, S. 2005. Perkembangan dan faktor-faktor yang mempengaruhi konversi lahan sawah serta dampak ekonominya. SOCA, Jurnal Sosial Ekonomi Pertanian5(2): 1-25.

IRRI. 2015. Final Evaluation of two IRRI projects (2012 to 2015). International Rice Research Institute. Manila - Philippines. p. 81.

Kleiber, D., Prigge, V.,  Melchinger, A.E., Burkard, F., Vicente, F.S., Palomino, G. & Gordillo, G.A. 2012. Haploid fertility in temperate and tropical maize germplasm. Crop Science 52: 623-630.

Lenaerts, B., Collard, B.C.Y. & Demont, M. 2019. Review: Improving global food security through accelerated plant breeding. Plant Science 287: 110207.

Mara, K.K.S., Purwoko, B.S., Sulistyono, E. & Dewi, I.S. 2015. Penampilan agronomi dan uji toleransi naungan galur dihaploid padi gogo hasil kultur antera. Jurnal Agronomi Indonesia 43(1): 1-7.

Mayakaduwa, D.M.R.G. & Silva, T.D. 2019. Flow cytometric detection of haploids, diploids and mixoploids among the anther-derived plants in indica rice (Oryza sativa L.). The Journal of Animal & Plant Sciences 29(5): 1344-1351.

Mishra, R., Narashima, G.J., Nageswara, R. & Kaushal, P. 2015. Development and characterization of elite doubled haploid lines from two indica rice hybrids. Rice Science 22(6): 290-299.

Mostafiz, B.S. & Wagiran, A. 2018. Efficient callus induction and regeneration in selected indica rice. Agronomy 8(5): 77.

Naik, N., Rout, P., Umakanta, N., Verma, R.L., Katara, J.L., Sahoo, K.K., Singh, O.N. & Samantaray, S. 2016. Development of doubled haploids from an elite indica rice hybrid (BS6444G) using anther culture. Plant Cell Tissue Organ Culture 128(3): 679-689.

Niazian, M. & Shariatpanahi, M.E. 2020. In vitro-based doubled haploid production: Recent improvements. Euphytica 216: 69.

Samantaray, S., Ali, J., Nicolas, K.L.C., Katara, J.L., Verma, R.L., Parameswaran, C., Devanna, B.N., Kumar, A., Dash, B. & Bhuyan, S.S. 2021. Doubled haploids in rice improvement: Approaches, applications, and future prospects. In Rice Improvement Physiological, Molecular Breeding and Genetic Perspectives, edited by Ali, J. & Wani, S.H. Springer. pp. 425-447.

Sasmita, P., Dewi, I.S. & Purwoko, B.S. 2001. Pengaruh generasi kalus terhadap regenerasi tanaman pada kultur antera padi (Oryza sativa L.) cv. Gajah Mungkur. Sain Teks (special edition). pp. 179-188.

Sasmita, P., Purwoko, B.S., Sujiprihati, S. & Somantri, I.H. 2002. Kultur antera padi gogo hasil persilangan kultivar dengan galur toleran naungan. Hayati 9(3): 89-93.

Seguí-Simarro, J.M. & Nuez, F. 2008a. Pathways to doubled haploidy: Chromosome doubling during androgenesis. Cytogenetic and Genome Research 120: 358-369.

Seguí-Simarro, J.M. & Nuez, F. 2008b. How microspore transform into haploid embyos: Changes associated with embryogenesis induction and microspore derived embryogenesis. Physiologia Plantarum 134(1): 1-12.

Septiningsih, E.M., Hidayatun, N., Sanchez, D.L., Nugraha, Y., Carandang, J., Pamplona, A.M., Collard, B.C.Y., Ismail, A.M. & Mackill. 2014. Accelerating the development of new submergence tolerant rice varieties: The case of Ciherang-Sub1 and PSB Rc18-Sub1. Euphytica 202: 259-268.

Safitri, H., Purwoko, B.S., Dewi, I.S. & Ardie, S.W. 2016. Kultur antera untuk mendapatkan galur padi toleran salinitas. Jurnal Agronomi Indonesia 44(3): 221-227.

Safitri, H., Purwoko, B.S., Wirnas, D., Dewi, I.S. & Abdullah, B. 2010. Daya kultur antera beberapa persilangan padi gogo dan padi tipe baru. Jurnal Agronomi Indonesia 38(2): 81-87.

Sexcion, F.S.H., Egdane, J.A., Ismail, A.M. & Dionisio-Sese, M.L. 2009. Morpho-physiological traits associated with tolerance of salinity during seedling stage in rice (Oryza sativa L.). Philippine Journal of Crop Science 34(2): 27-37.

Takeda, S. & Matsuoka, M. 2008. Genetic approaches to crop improvement: Responding to environmental and population changes. Nature Reviews Genetics 9: 444-457.

Talebi, R., Rahemi, M.R., Arefi, H., Nourozi, M. & Bagheri, N. 2007. In vitro plant regeneration through anther culture of some Iranian local rice (Oryza sativa L.) cultivars. Pakistan Journal of Biological Sciences 10: 2056-2060.

Tripathy, S.K., Swain, D., Mohapatra, P.M., Prusti, A.M., Sahoo, B., Panda, S., Dash, M., Chakma, B. & Behera, S.K. 2019. Exploring factors affecting anther culture in rice (Oryza sativa L.). Journal of Applied Biology & Biotechnology 7(2): 87-92.

Uga, Y., Okuno, K. & Yano, M. 2011. Dro1, a major QTL involved in deep rooting of rice under upland field conditions. Journal of Experimental Botany 62(8): 2485-2494.

Wening, R.H. 2020. Genetic and morphophysiological studies of rice tolerance to submergence and drought stresses to support breeding multi tolerant varieties.  IPB University, Ph.D. Thesis (Unpublished).

Yamagishi, M., Otani, M., Higashi, M., Fukuta, Y., Fukui, K. & Shimada, T. 1998. Chromosomal regions controlling anther culturability in rice (Oryza sativa L.). Euphytica 103: 227-234.

Yorobe, J.M., Ali, J., Pede, V.O., Rejesusd, R.M. & Velarde, O.P. 2016. Yield and income effects of rice varieties with tolerance of multiple abiotic stresses: The case of green super rice (GSR) and fooding in the Philippines. Agricultural Economic 47: 261-271.

Zhang, Z.H. 1989. The practicability of anther culture breeding in rice. In Review of Advances in Plant Biotechnology, 1985-88, edited by Mujeeb-Kazi, A. & Stich, L.A. Los Baños, Philippines, International Maize and Wheat Improvement Center, International Rice Research Institute. pp. 31-42.

Zhou, H. 1996. Genetics of green plantlet regeneration from anther culture of cereals. In In Vitro Haploid Production in Higher Plants. Current Plant Science and Biotechnology in Agriculture. Vol. 24, edited by Jain, S.M., Sopory, S.K. & Veilleux, R.E. Dordrecht: Springer. pp. 169-187.

^*Pengarang untuk surat-menyurat; email: bspurwoko@apps.ipb.ac.id