CRISPR/Cas9-mediated mutation of OsSWEET14 in rice cv. Zhonghua11 confers resistance to Xanthomonas oryzae pv. oryzae without yield penalty
CRISPR/Cas9-mediated mutation of OsSWEET14 in rice cv. Zhonghua11 confers resistance to Xanthomonas oryzae pv. oryzae without yield penalty
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DOI：
10.1186/s12870-020-02524-y
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发表时间：
2020-07
影响因子：
5.3
通讯作者：
Xuan Zeng;Yufen Luo;Nga T. T. Vu-Nga-T.-T.-Vu-49168727;Shujuan Shen;Kuai-fei Xia;Mingyong Zhang
中科院分区：
文献类型：
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作者：
Xuan Zeng;Yufen Luo;Nga T. T. Vu-Nga-T.-T.-Vu-49168727;Shujuan Shen;Kuai-fei Xia;Mingyong Zhang
水稻白叶枯病是由水稻白叶枯病菌引起的。水稻白叶枯病(Xoo)是东南亚和西非的一种毁灭性水稻病害。OsSWEET14编码一种糖转运蛋白，是白叶枯病的主要易感基因，可被来自亚洲或非洲Xoo菌株的四种不同的转录激活物样(TAL)效应分子靶向。然而，Kitaake背景中的OsSWEET14单基因敲除或启动子突变对非洲Xoo菌株具有中等抗性甚至敏感。因此，在本研究中，我们在水稻品种中敲除了OsSWEET14。中华民国11日疾病背景评估。在本研究中，我们利用CRISPR/Cas9通过改变水稻新品种基因组中OsSWEET14的相应编码区来干扰其功能。中华11号(CR-S14)。我们总共获得了9个不同的OsSWEET14突变等位基因。除了对亚洲Xoo株具有广谱抗性外，被测试的突变等位基因对非洲Xoo株AXO1947也表现出很强的抗性。此外，在茎、叶鞘、叶片和根等维管组织中检测到了OsSWEET14的表达。OsSWEET14的中断导致了株高的增加，而产量却没有下降。OsSWEET14在中华11号背景上的突变能够赋予对非洲Xoo株系AXO1947和亚洲Xoo株系PXO86很强的抗性。在正常生长条件下，CR-S14生殖生长正常，株高增加。这些结果表明，在Kitaake背景下，CR-S14可能比Sweet14单基因敲除突变体更适合作为水稻疫病抗性诊断试剂盒。在利用OsSWEET14进行抗病水稻育种时，需要考虑遗传背景和株高的增加。
Bacterial blight of rice, caused by Xanthomonas oryzae pv. oryzae (Xoo), is a devastating rice disease in Southeast Asia and West Africa. OsSWEET14, encoding a sugar transporter, is known to be a major susceptible gene of bacterial blight targeted by four different transcription activator-like (TAL) effectors from either Asian or African Xoo strains. However, the OsSWEET14 single knockout or promoter mutants in the Kitaake background are moderately resistant or even susceptible to African Xoo strains. Therefore, in this study, we knocked out OsSWEET14 in rice cv. Zhonghua 11 background for disease assessment. In this study, CRISPR/Cas9 was utilized to disrupt the function of OsSWEET14 by modifying its corresponding coding region in the genome of rice cv. Zhonghua 11 (CR-S14). In total, we obtained nine different OsSWEET14-mutant alleles. Besides conferring broad-spectrum resistance to Asian Xoo strains, tested mutant alleles also showed strong resistance to African Xoo strain AXO1947. Moreover, the expression of OsSWEET14 was detected in vascular tissues, including the stem, leaf sheath, leaf blade and root. The disruption of OsSWEET14 led to increased plant height without a reduction in yield. Disruption of OsSWEET14 in the Zhonghua 11 background is able to confer strong resistance to African Xoo strain AXO1947 and Asian Xoo strain PXO86. CR-S14 has normal reproductive growth and enhanced plant height under normal growth conditions. These results imply that CR-S14 may serve as a better tester line than sweet14 single-knockout mutant in the Kitaake background for the diagnostic kit for rice blight resistance. The genetic background and increased plant height need to be taken into consideration when utilizing OsSWEET14 for resistant rice breeding.