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


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.


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



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)


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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

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