Chinese Journal OF Rice Science >
Creation of Low-Cd-accumulating indica Rice by Disruption of OsNramp5 Gene via CRISPR/Cas9
Received date: 2019-03-05
Revised date: 2019-05-24
Online published: 2019-09-10
【Objective】 To accelerate the breeding process of low-Cd-accumulating rice,【Methods】 the widely applied indica pure-line cultivars, Wushansimiao and Zhongzao 35, and the popular parents of hybrids, Huazhan and Wufeng B were used as materials to create OsNramp5 knockout lines via CRISPR/Cas9. The marker-free OsNramp5 mutants together with the wild types were grown either in a Cd-contaminated field (Field A) or in a non-Cd-contaminated field (Field B) for subsequent experiments. The changes of the grain Cd content were investigated for plants grown in both Field A and Field B. The changes of the contents of other related minerals were also examined but only for those plants grown in Field A. The agronomic changes of the mutants were further assessed by examining those plants grown in Field B. 【Results】 Rice lines with mutations at OsNramp5 were successfully generated for all the four cultivars. On one hand, the grain Cd level in OsNramp5 mutants grown in Field B was reduced by 85.5% on average (<0.02 mg/kg); as a result of OsNramp5 mutation, the Cd level in plants grown in Field A, was reduced by 94.8% (<0.1 mg/kg), meanwhile, the Mn level was decreased by 52.7%; on the contrary, the Cr level was elevated by 59.5% on average for the four cultivars and an increase of 79.1% in the Pb level was also observed but only in OsNramp5 mutants of Huazhan; the levels of other mineral elements including Fe, Cu , Zn, Ca, Se and As were slightly or hardly affected. On the other hand, a slight decrease in plant height and an average yield loss of 6.9% were observed due to OsNramp5 mutation for all cultivars; the yield loss could be ascribed to a reduction in seed setting rate and seed weight and was compensated by a slight increase in effective tiller number. 【Conclusion】 Low-Cd-accumulating rice plants can be generated by OsNramp5 disruption at the cost of an acceptable yield loss and the low-Cd-accumulating rice lines produced in the present study may serve as promising cultivars to eliminate rice Cd contamination in many areas.
Key words： indica rice; cadmium; genetic editing; breeding
Qizhang LONG, Yonglan HUANG, Xiuying TANG, Huimin WANG, Ming LU, Linfeng YUAN, Jianlin WAN . Creation of Low-Cd-accumulating indica Rice by Disruption of OsNramp5 Gene via CRISPR/Cas9[J]. Chinese Journal OF Rice Science, 2019 , 33(5) : 407 -420 . DOI: 10.16819/j.1001-7216.2019.9026
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