Chinese Journal OF Rice Science >
Evaluation of New japonica Rice Lines with Low Cadmium Accumulation and Good Quality Generated by Knocking Out OsNramp5
Received date: 2022-05-06
Revised date: 2022-07-31
Online published: 2023-01-10
【Objective】 It is crucial to elucidate the effects of OsNramp5 gene knockout on cadmium and other metal elements accumulation, yield and quality in japonica rice, which can provide new materials and theoretical references for scientific and efficient production of high-quality and green japonica rice.【Methods】 Using CRISPR/Cas9 technology, the OsNramp5 was knocked out under the background of Nanjing 46 (NJ46), a japonica rice variety with good taste and high yield. Combined with molecular marker-assisted selection, some OsNramp5 knockout lines without exogenous genes were obtained. The changes in yield and quality traits and seven metal elements contents such as cadmium, manganese and selenium in grains after spraying zinc fertilizer or selenium fertilizer and cultivated in soils with different cadmium content were investigated. 【Results】 Five transgenic plants with different mutations of OsNramp5 were obtained in the T0 generation, and specific molecular markers of three of them were designed. Then, combined with marker-assisted selection, homozygous knockout lines without hygromycin selective marker gene and CAS9 protein gene were obtained in the T2 generation of each plant. Compared with the wild type, the cadmium content in the whole plant including the grain of the knockout lines and the manganese content in shoots were significantly lower than those of the wild type in both high and low concentrations of cadmium soil. The plant height and grain number per panicle of the knockout lines decreased slightly, and the yield per plant decreased significantly in the knockout lines at high and medium tillering fertilizer levels while no changes were observed at the low tillering fertilizer level. The appearance of the knockout lines was better than that of the control, and the contents of protein and amylose were significantly higher than those of the control. The contents of copper, manganese and selenium in grains of knockout lines were significantly lower than those of the control NJ46, but zinc and selenium fertilizer could significantly increase the contents of zinc and selenium, and further reduce the content of cadmium in grains. Application of selenium was beneficial to rice appearance quality, but application of zinc was contrary. Spraying both fertilizers reduced the edible value of rice, worsening taste quality. The change of seed setting rate of the knockout lines under extreme high and low temperature conditions was the same as that of the control, but the resistance level to rice sheath blight was significantly reduced.【Conclusion】 We obtained marker-free OsNramp5 knockout lines under the background of japonica variety with high quality and yield, and proved that OsNramp5 was of great value in the production of low-cadmium, Se- and zinc-rich, though their yield and eating quality were decreased to different extents.
PEI Feng, WANG Guangda, GAO Peng, FENG Zhiming, HU Keming, CHEN Zongxiang, CHEN Hongqi, CUI Ao, ZUO Shimin . Evaluation of New japonica Rice Lines with Low Cadmium Accumulation and Good Quality Generated by Knocking Out OsNramp5[J]. Chinese Journal OF Rice Science, 2023 , 37(1) : 16 -28 . DOI: 10.16819/j.1001-7216.2023.220503
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