摘要
水稻颖壳与糙米颜色作为重要的农艺性状,长期以来在水稻育种及遗传学研究领域中被广泛应用。本研究从尼泊尔稻种资源中鉴定到一份稻穗及糙米均呈现金黄色的籼稻材料‘LAL SAR’,遗传分析表明上述表型受单隐性核基因控制,将其命名为gpr1(golden panicle and brown rice 1)。利用图位克隆技术将目标基因精细定位于水稻第3号染色体上47 kb的区间内,并确定其候选基因为LOC_Os03g60509。该基因编码查尔酮异构酶(chalcone isomerase, CHI),是黄酮类化合物生物合成途径中的关键酶。PCR扩增测序和RT-PCR表达分析发现,在‘LAL SAR’中gpr1基因的启动子区存在大片段插入,导致该基因表达缺失。利用CRISPR/Cas9技术在粳稻品种‘中花11’(‘ZH11’)中敲除了gpr1的显性等位基因GPR1,敲除植株的稻穗及糙米均呈现出金黄色,并且其颖壳中的柚皮素查尔酮的含量显著提高。研究结果表明GPR1基因参与水稻类黄酮的生物合成,本研究为提升水稻品质与遗传改良提供了重要的理论依据与基因资源,具有重要的应用价值。
The colors of panicle hull and brown rice are crucial agronomic traits in rice,which are widely used in breeding and genetic research.Here,a rice variety named‘LAL SAR’characterized by golden panicle and brown rice was identified from Nepal.Genetic analysis revealed that these phenotypes were controlled by a single recessive nuclear gene,which was designated as gpr1(golden panicle and brown rice 1).Using the strategy of map-based cloning,we mapped gpr1 into a 47 kb interval on the rice chromosome 3 and identified its candidate gene as LOC_Os03g60509.This gene encodes a chalcone isomerase,a key enzyme in the flavonoid biosynthetic pathway.Through PCR sequencing and RT-PCR analysis,a long-fragment insertion was found in the promoter of gpr1 in‘LAL SAR’,which completely suppressed the expression of this gene.Using CRISPR/Cas9 technology,we successfully knocked out GPR1(the dominant allele of gpr1)from a Oryza sativa subsp.japonica variety‘Zhonghua 11’.The knockout mutant plants exhibited golden panicles and brown rice,with significantly increased naringenin chalcone content in the hull.Our results indicate that GPR1 participates in the flavonoid biosynthesis,providing a critical theoretical foundation and a gene resource for rice quality improvement and genetic enhancement.
作者
季永再
卞晓波
冯志状
李范晶
胡卓尔
胡瑶洁
刘鹏程
马伯军
陈析丰
JI Yongzai;BIAN Xiaobo;FENG Zhizhuang;LI Fanjing;HU Zhuo’er;HU Yaojie;LIU Pengcheng;MA Bojun;CHEN Xifeng(College of Life Sciences,Zhejiang Normal University,Jinhua 321004,Zhejiang,China;Jinhua Academy of Agricultural Sciences,Jinhua 321004,Zhejiang,China;Key Laboratory for Innovative Utilization of Characteristic Food Crop Resources in Central Zhejiang,Jinhua 321004,Zhejiang,China)
出处
《生物工程学报》
北大核心
2026年第1期356-366,共11页
Chinese Journal of Biotechnology
基金
国家自然科学基金(32272096)
浙江省自然科学基金(LZ24C130003)
浙江省大学生科技创新活动计划(新苗人才计划)(2023R404039)
金华市重点科技计划(2024-2-017,2023-2-014)。
