Advertisement

Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 139, Issue 3, pp 577–587 | Cite as

In vitro induction of auto-allotetraploid in a newly developed wild rice line from Oryza alta Swallen

  • Liusheng Zhang
  • Fimanekeni Ndaitavela Shivute
  • Muhammad Qasim Shahid
  • Nabieu Kamara
  • Jinwen Wu
  • Xiangdong LiuEmail author
Original Article
  • 116 Downloads

Abstract

Oryza alta Swallen is an important germplasm for rice resistance breeding; however, its CCDD genome (2n = 48) resulted in low crossability when the wild rice was crossed with O. sativa and restricted the success of transferring the desirable traits into cultivated rice. Induction of polyploidy is an efficient way for overcoming the low crossability among different species. A new O. alta line, Huaye 5, was developed by our group in 2016, which had high fertility (64.93%) and photoperiod-insensitive. Huaye 5 was used to induce auto-allotetraploidy using tissue culture in the present study. The tissue culture system was established by comparing five basic media (N6, B5, MS, NB and MB), two hormones (2,4-D and 6-BA) for induction and two differentiation media (MS and NB), and then induced auto-allotetraploid in the wild rice line by colchicine. The medium and hormone combinations of NB + 2,4-D (2.5 mg/L) + 6-BA (1.0 mg/L) produced the induction rate of 20%, and MS medium was found to be a suitable medium for callus induction with a differentiation rate of 10.15%, and the treatment of 600 mg/L colchicine for 24 h was the best protocol for inducing auto-allotetraploid. Subsequently, auto-allotetraploid plants (2n = 96) were obtained in the present study and their ploidy levels were detected by using flow cytometry, stomata size and chromosomes count methods. Many inclusions in the parenchyma cells surrounding vascular bundle were observed in auto-allotetraploid rice compared to the parent. We developed a new germplasm from O. alta, and established a protocol of in vitro induction of auto-allotetraploid, which can be used for crossing with autotetraploid rice.

Key message

We have successfully established a protocol for the in vitro induction of auto-allotetraploid in Oryza alta, which can be used to cross with autotetraploid or neo-tetraploid rice.

Keywords

Rice (Oryza sativaO. alta Swallen Allotetraploid Huaye 5 Tissue culture 

Notes

Acknowledgements

The authors thank Ms. Shuhong Yu, Yiwen Wang, Wu Jin, Wenchun Chen, and other lab members for assistance. This work was supported by the Guangzhou Science and Technology Key Program to XD Liu (201707020015), NSFC to XD Liu (31571625) and Opening Foundation of Guangdong Province Key Laboratory of Plant Molecular Breeding (GPKLPMB201803).

Author contributions

XDL conceived and designed the experiments. FNS, XDL, MQS, LSZ and NK wrote the paper. LSZ, FNS, MQS and JWW performed the experiment and analyzed the data. XDL and JWW developed Huaye 5. All authors read and approved the final version of manuscript.

Compliance with ethical standards

Conflict of interest

The authors declare that there are no conflicts of interest.

Supplementary material

11240_2019_1701_MOESM1_ESM.pdf (1.3 mb)
Supplementary material 1 (PDF 1322 kb)

References

  1. Abe T, Futsuhara Y (1986) Genotypic variability for callus formation and plant generation in rice (Oryza sativa L.). Theor Appl Genet 72(1):3–10PubMedGoogle Scholar
  2. Ai SS, Li JR, Zeng XF, Zhao DG (2012) Establishment of the tissue culture and regeneration system of Guizhou rice landrace. Mol Plant Breeding 10(4):469–475Google Scholar
  3. Aleza P, Juarez J, Cuenca J, Ollitrault P, Navarro L (2010) Recovery of citrus triploid hybrids by embryo rescue and flow cytometry from 2x × 2x sexual hybridisation and its application to extensive breeding programs. Plant Cell Rep 29(9):1023–1034PubMedGoogle Scholar
  4. Bayliss MW (1973) Origin of chromosome number variation in cultured plant cells. Nature 246:529–530Google Scholar
  5. Beck SL, Dunlop RW, Fossey A (2003) Stomatal length and frequency as a measure of ploidy level in black wattle, Acacia mearnsii (de Wild). Bot J Linn Soc 41:177–181Google Scholar
  6. Bouvier L, Guerif PH, Djulbic M, Durel CHE, Chevreau E, Lespinase Y (2002) Chromosome doubling of pear haploid plants and homozygosity assessment using isozyme and microsatellite markers. Euphytica 123(2):255–262Google Scholar
  7. Chen C, Hou X, Zhang H, Wang G, Tian L (2011) Induction of Anthurium andraeanum “Arizona’’ tetraploid by colchicine in vitro. Euphytica 181(2):137–145Google Scholar
  8. Chen L, Shahid MQ, Wu J, Chen Z, Wang L, Liu X (2018) Cytological and transcriptome analyses reveal abrupt gene expression for meiosis and saccharide metabolisms that associated with pollen abortion in autotetraploid rice. Mol Genet Genomics 293:1407–1420PubMedPubMedCentralGoogle Scholar
  9. Chen L, Yuan Y, Wu J, Chen Z, Wang L, Shahid MQ, Liu X (2019) Carbohydrate metabolism and fertility related genes high expression levels promote heterosis in autotetraploid rice harboring double neutral genes. Rice 12:34PubMedPubMedCentralGoogle Scholar
  10. Cheng ZM, Korban SS (2011) In vitro ploidy manipulation in the genomics era. Plant Cell Tissue Organ Cult 104(3):281–282Google Scholar
  11. Dolezel J (1997) Applications of flow cytometry for the study of plant genomes. J Appl Genet Poland 38(3):285–302Google Scholar
  12. Fu XL, Lu YG, Liu XD, Li JQ, Feng JH (2007) Cytological mechanisms of interspecific incrossability and hybrid sterility between Oryza sativa L. and O. alta Swallen. Chin Sci Bull 52(6):755–765Google Scholar
  13. Fu XL, Lu YG, Liu XD, Li JQ, Zhao XJ (2011) Cytological behavior of hybridization barriers between Oryza sativa and Oryza officinalis. Agric Sci China 10(10):1489–1500Google Scholar
  14. Gireesh C (2018) Oryza alta Swallen. In: Mondal T, Henry R (eds) The Wild Oryza Genomes Compendium of Plant Genomes. Springer, Cham, pp 55–59.  https://doi.org/10.1007/978-3-319-71997-9_4 CrossRefGoogle Scholar
  15. Guo H, Mendrikahy JN, Xie L, Deng J, Lu Z, Wu J, Li X, Shahid MQ, Liu X (2017) Transcriptome analysis of neo-tetraploid rice reveals specific differential gene expressions associated with fertility and heterosis. Sci Rep 7:40139PubMedPubMedCentralGoogle Scholar
  16. He J, Shahid MQ, Li Y, Guo H, Cheng X, Liu X, Lu Y (2011) Allelic interaction of F1 pollen sterility loci and abnormal chromosome behaviour caused pollen sterility in intersubspecific autotetraploid rice hybrids. J Exp Bot 62(13):4433–4445PubMedPubMedCentralGoogle Scholar
  17. Huang HP, Gao SL, Wang DL, Huang P (2014) Autotetraploidy induced in Nianmaohuangqin (Radix Scutellariae viscidulae) with colchicine in vitro. J Tradit Chin Med 34(2):199–205.  https://doi.org/10.1016/S0254-6272(14)60079-0 CrossRefPubMedGoogle Scholar
  18. Huang LJ, Wang H, Shahid MQ, Zeng B (2019) Chlorothalonil: an effective bacteriostatic agent for bud induction of Acacia auriculiformis under open condition (non-axenic). Plant Methods 15:5PubMedPubMedCentralGoogle Scholar
  19. Jena KK, Ballesfin ML, Vinarao RB (2016) Development of Oryza sativa L. by Oryza punctata kotschy ex steud. monosomic addition lines with high value traits by interspecific hybridization. Theor Appl Genet 129(10):1873–1886.  https://doi.org/10.1007/s00122-016-2745-8 CrossRefPubMedGoogle Scholar
  20. Jin J, Li SQ, Xie HW, Li NW, Huang WC, Hu J, Wang K, Zhu RS, Zhu YG (2013) Germplasm development, discovery and utilization of valuable genes from wild relatives of Oryza genus. J Wuhan Univ (Nat Sci Ed) 59(1):10–16 (in Chinese)Google Scholar
  21. Kausch AP, Tilelli M, Hague J, Heffelfinger C, Cunha D, Moreno M, Dellaporta SL, Nelson L (2016) In situ embryo rescue for generation of wide intra and interspecific hybrids of Panicum virgatum L. Plant Biotechnol J 14(11):2168–2175PubMedPubMedCentralGoogle Scholar
  22. Li YJ, Chen YL, Wang LL, Zhao SM, Zhu ZG (2005) Effects of 2,4-D and 6-BA on callus culture of rice. J Hebei Norm Uni 29(4):395–398Google Scholar
  23. Li T, Chen J, Qiu S, Zhang Y, Wang P, Yang L, Lu Y, Shi J (2012) Deep sequencing and microarray hybridization identify conserved and species-specific microRNAs during somatic embryogenesis in hybrid yellow poplar. PLoS ONE 7(8):e43451Google Scholar
  24. Li X, Shahid MQ, Wu J, Wang L, Liu X, Lu Y (2016) Comparative small RNA analysis of pollen development in autotetraploid and diploid rice. Int J Mol Sci 17(4):499PubMedPubMedCentralGoogle Scholar
  25. Li X, Shahid MQ, Xia J, Lu Z, Fang N, Wang L, Wu J, Chen Z, Liu X (2017) Analysis of small RNAs revealed differential expressions during pollen and embryo sac development in autotetraploid rice. BMC Genomics 18:129PubMedPubMedCentralGoogle Scholar
  26. Li X, Yu H, Jiao Y, Shahid MQ, Wu J, Liu X (2018) Genome-wide analysis of DNA polymorphisms, the methylome and transcriptome revealed that multiple factors are associated with low pollen fertility in autotetraploid rice. PLoS ONE 13(8):e0201854PubMedPubMedCentralGoogle Scholar
  27. Liang YT, Pan YH, Yu JP (2014) Study on callus induction and differentiation of O alta wild rice. Southwest China J Agri Sci 27(3):905–909Google Scholar
  28. Liu WS, Liu DC, Feng CJ, Zhang AM, Li SH (2006) Genetic diversity and phylogenetic relationships in plum germplasm resources revealed by RAPD markers. Hort Sci Biotechnol 81(2):242–250Google Scholar
  29. Liu YH, Yu L, Cui TC (2010) Study on callus induction and plant regeneration of zoysiagrass. Pratacult Sci 27(03):107–111Google Scholar
  30. Ma Y, Xue H, Zhang L, Zhang F, Ou CQ, Wang F, Zhang ZH (2016) Involvement of auxin and brassinosteroid in dwarfism of autotetraploid apple (Malus × domestica). Sci Rep 6:26719.  https://doi.org/10.1038/srep26719 CrossRefPubMedPubMedCentralGoogle Scholar
  31. Mao L, Zhu LH, Zhou Q, Wang XP, Hu H (1995) RFLP analysis of the progeny from Oryza alta Swallen × Oryza sativa L. Genome 38(5):913–918PubMedGoogle Scholar
  32. Mizobuchi R, Fukuoka S, Tsushima S, Yano M, Sato H (2016) QTLs for resistance to major rice diseases exacerbated by global warming: brown spot, bacterial seedling rot and bacterial grain rot. Rice 9:23Google Scholar
  33. Mustafa NS, Liu F, Odongo MR, Wang XX, Cai XY, Zhang ZM, Wang KB (2017) Effects of colchicine treatments on chromosome doubling in three diploid cotton Species. J Multidiscip Engi Sci Techol (JMEST) 4(4):7140–7145Google Scholar
  34. Ni WJ, Cheng FM, Zhang JJ (2007) Gaining regeneration plantlets from embryoids in leaf tissue culture of Oryza alta L. Shanghai J Agri Sci 23(4):18–20 (in Chinese)Google Scholar
  35. Nilanthi D, Chen XL, Zhao FC, Yang YS, Wu H (2009) Induction of tetraploids from petiole explants through colchicine treatments in Echinacea purpurea L. J Biomed Biotechnol 2009:343485PubMedPubMedCentralGoogle Scholar
  36. Parera PCD, Dahanayake N (2016) Chromosome observation of rice root tip and effect of callus colour variation and texture in different colchicine concentrations on the induction of rice polyploids. Univers J Agric Res 4(2):56–59Google Scholar
  37. Qiao CZ, Wu MS, Dai FB, Cui X, Li L (1989) Studies on polyploid breeding of Isatis indigotica Fort. Acta Bot Sin 31(9):678–683 (in Chinese)Google Scholar
  38. Qin XY, Zhu RC, Tang JH, Li WK, Li DY, Wei SM, Huang FK (2007) Genetic analysis and utilization of brown planthopper (BPH)-resistant genes in Oryza officinalis Wall. J Plant Genet Res 8(1):41–45 (in Chinese) Google Scholar
  39. Rauf S, Munir H, Abdullojon E, Basra SM (2006) Role of colchicine and plant growth regulators to overcome interspecific incompatibility. Gen Appl Plant Physiol 32(3–4):223–232Google Scholar
  40. Shahid MQ, Sun J, Wei C, Zhang P, Liu X (2010) Studies on the abnormality of embryo sac and pollen fertility in autotetraploid rice during different growing seasons. Pak J Bot 42(1):7–19Google Scholar
  41. Shahid MQ, Liu G, Li J, Naeem M, Liu X (2011) Heterosis and gene action study of agronomic traits in diploid and autotetraploid rice. Acta Agric Scand Sect B 61(1):23–32.  https://doi.org/10.1080/09064710903428140 CrossRefGoogle Scholar
  42. Shahid MQ, Xu H, Lin S, Chen Z, Naeem M, Li Y, Liu X (2012) Genetic analysis and hybrid vigor study of grain yield and other quantitative traits in autotetraploid rice. Pak J Bot 44(1):237–246Google Scholar
  43. Shahid MQ, Li Y, Saleem MF, Naeem M, Wei C, Liu X (2013) Yield and yield components in autotetraploid and diploid rice genotypes (indica and japonica) sown in early and late seasons. Aust J Crop Sci 7(5):632–641Google Scholar
  44. Speckmann G, Post J, Dijkstra H (1965) The length of stomata as an indicator for polyploidy in rye-grasses. Euphytica 14(3):225–230Google Scholar
  45. Sung J, Lee S, Chung JW, Edwards G, Ryu H, Kim T (2017) Photosynthesis, metabolite composition and anatomical structure of Oryza sativa and two wild relatives, O. grandiglumis and O. alta. Rice Sci 24(4):218–227Google Scholar
  46. Urwin NAR (2014) Generation and characterisation of colchicine-induced polyploid Lavandula x intermedia. Euphytica 197(3):331–339Google Scholar
  47. Vaughan AD (1994) The wild relatives of rice: a genetic resource handbook. IRRI Publication, Los Baños, p 147Google Scholar
  48. Veasey AE, Karasawa MG, Santos PP, Rosa MS, Mamani E, Oliveira GCX (2004) Variation in the loss of seed dormancy during after-ripening of wild and cultivated rice species. Ann Bot 94(6):875–882PubMedPubMedCentralGoogle Scholar
  49. Wang AY, Chen DL, Cai DT (2005) Application of distant hybridization and allopolyploidy in rice breeding. Chin J Plant Sci 23(5):491–495Google Scholar
  50. Wang D, Wang Y, Zhao H (2010) Comparative analysis of genomes of high wild rice and wide-leaf wild rice using C_0t-1 DNA. Chin J Rice Sci 24(2):131–136Google Scholar
  51. Wei YX, Zhu MZ, Qiao HY, Li F, Zhang SJ, Zhang SF, Zhang H, Sun RF (2018) Characterization of interspecific hybrids between flowering Chinese cabbage and broccoli. Sci Hortic 240:552–557Google Scholar
  52. Wu J, Shahid MQ, Guo H, Yin W, Chen Z, Wang L, Liu X, Lu Y (2014) Comparative cytological and transcriptomic analysis of pollen development in autotetraploid and diploid rice. Plant Reprod 27:181–196PubMedGoogle Scholar
  53. Wu J, Shahid MQ, Chen L, Chen Z, Wang L, Liu X, Lu Y (2015) Polyploidy enhances F1 pollen sterility loci interactions that increase meiosis abnormalities and pollen sterility in autotetraploid rice. Plant Physiol 169:2700–2717PubMedPubMedCentralGoogle Scholar
  54. Wu JW, Chen L, Shahid MQ, Chen MY, Dong QL, Li JR, Xu XS, Liu XD (2017) Pervasive interactions of Sa and Sb loci cause high pollen sterility and abrupt changes in gene expression during meiosis that could be overcome by double neutral genes in autotetraploid rice. Rice 10:49PubMedPubMedCentralGoogle Scholar
  55. Xing SH, Guo XB, Wang Q, Pan QF, Tian YS, Liu P, Zhao JY, Wang GF, Sun XF, Tang KX (2011) Induction and flow cytometry identification of tetraploids from seed-derived explants through colchicine treatments in Catharanthus roseus (L.) G. Don. J Biomed Biotechnol 2011:793198PubMedPubMedCentralGoogle Scholar
  56. Xiong YG, Gan L, Hu YP, Sun WC, Zhou X, Song ZJ, Zhang XH, Li Y, Yang ZF, Xu WF, Zhang JH, He YC, Cai DT (2019) OsMND1 regulates early meiosis and improves the seed set rate in polyploid rice. Plant Growth Regul 87:341–356Google Scholar
  57. Xu L, Najeeb U, Naeem MS, Daud MK, Cao JS, Gong HJ, Shen WQ, Zhou WJ (2011) Induction of tetraploidy in Juncus effusus by colchicine. Biol Plant 54(4):659–663Google Scholar
  58. Yamaki S, Ohyanagi H, Yamasaki M, Eiguchi M, Miyabayashi T, Kubo T, Kurata N, Nonomura KI (2013) Development of INDEL markers to discriminate all genome types rapidly in the genus Oryza. Breed Sci 63(3):246–254PubMedPubMedCentralGoogle Scholar
  59. Yan HH, Hu HY, Fu Q, Yu HY, Tang SX, Xiong ZM, Min SK (1996) Morphological and cytogenetical studies of the hybrids between Oryza sativa and Oryza officinalis. Chin J Rice Sci 10(3):138–142 (in Chinese) Google Scholar
  60. Yang PM, Huang QC, Qin GY, Zhao SP, Zhou JG (2014) Different drought-stress responses in photosynthesis and reactive oxygen metabolism between autotetraploid and diploid rice. Photosynthetica 52(2):193–202.  https://doi.org/10.1007/s11099-014-0020-2 CrossRefGoogle Scholar
  61. Yi CD, Tang SZ, Zhou Y, Liang GH, Gong ZY, Gu MH (2008) Development and characterization of interspecific hybrids between Oryza sativa and O. latifolia by in situ hybridization. Chin Sci Bull 53(19):2973–2980Google Scholar
  62. Zhang J, Zhang M, Deng X (2007) Obtaining autotetraploids in vitro at a high frequency in Citrus sinensis. Plant Cell Tissue Organ Cult 89(2):211–216Google Scholar
  63. Zhang QY, Luo FX, Liu L, Guo FC (2010) In vitro induction of tetraploids in crape myrtle (Lagerstroemia indica L.). Plant Cell Tissue Organ Cult 101(1):41–47Google Scholar
  64. Zhang XH, Wang AY, Du ZJ, Wang WW, He YC, Cai DT (2014) An efficient method of developing synthetic allopolyploid rice (Oryza spp.). Genet Resour Crop Evol 61(4):809–816Google Scholar
  65. Zhao W, Xia HB, Zhang SJ, Cai XX, Lu BR (2008) Differentiation of Oryza species revealed by the indica-japonica specific insertion/deletion (InDel) molecular markers. J Fudan Univ (Nat Sci) 47(3):281–287 (in Chinese) Google Scholar
  66. Zhou HW, Zeng WD, Yan HB (2017) In vitro induction of tetraploids in cassava variety ‘Xinxuan 048’ using colchicine. Plant Cell Tissue Org 128(3):723–729Google Scholar

Copyright information

© Springer Nature B.V. 2019

Authors and Affiliations

  1. 1.State Key Laboratory for Conservation and Utilization of Subtropical Agro-BioresourcesSouth China Agricultural UniversityGuangzhouChina
  2. 2.Guangdong Provincial Key Laboratory of Plant Molecular BreedingSouth China Agricultural UniversityGuangzhouChina
  3. 3.College of AgricultureSouth China Agricultural UniversityGuangzhouChina

Personalised recommendations