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| 1 | Improved Base Editor for Efficiently Inducing Genetic Variations in Rice with CRISPR/Cas9- Guided Hyperactive hAID Mutant显示文摘 | Bin Ren Fang Yan Yongjie Kuang Na Li Dawei Zhang Xueping Zhou Honghui Lin Huanbin Zhou | 2018 | Molecular Plant2018，11，4： | 46 |
| 2 | Base-Editing-Mediated Artificial Evolution of OsALS1 In Planta to Develop Novel Herbicide-Tolerant Rice Germplasms显示文摘Recently developed CRISPR-mediated base editors,which enable the generation of num erous nucleotide changes in target genomic regions,have been widely adopted for gene correction and generation of crop germ plasms containing im portant gain-of-function genetic variations.How ever,to engineer target genes with unknown functional SNPs remains challenging.To address this issue,we present here abase-e diting-mediated gene evolution(BEMGE)m ethod,employing both Cas9n-based cytosine and adenine base editors as well as a single-guide RNA(sgRNA)library tiling the full-length coding region,for developing novel rice germ plasm swith mutations in any endogenous gene.To this end,OsALS1 was artificially evolved in rice cells using BEMGE through both Agrobacterium-mediated and particle-bom bardment-mediated transform ation.Four different types of amino acid substitutions in the evolved OsALS1,derived from two sites that have never been targeted by natural or human selection during rice dom estication,were identified,conferring varying levels of tolerance to the herbicide bispyribac-sodium.Furtherm ore,the P171F substitution identified in a strong OsALS1 allele was quickly introduced into the commercial rice cultivar Nangeng 46 through precise base editing w ith the corresponding base editor and sgRNA.Collectively,these data indicate great potential of BEMGE in creating important genetic variants of target genes for crop improvement. | Yongjie Kuang Shaofang Li Bin Ren Fang Yan Carl Spetz Xiangju Li Xueping Zhou Huanbin Zhou | 2020 | Molecular Plant2020，13，4： | 37 |
| 3 | A CRISPR/Cas9 toolkit for efficient targeted base editing to induce genetic variations in rice显示文摘Dear Editor,Gene-targeting technologies using sequence specific nucleases have been widely adopted to induce genetic modifications in both plant molecular biology research and crop improvement,of which generating targeted point mutation for gain-of-function phenotype is of great value.Although tremendous efforts have been made in developing geneinsertion or replacement approaches by | Bin Ren Fang Yan Yongjie Kuang Na Li Dawei Zhang Honghui Lin Huanbin Zhou | 2017 | Science China(Life Sciences)2017，60，5： | 28 |
| 4 | Highly Efficient A. T to G. C Base Editing by Cas9n- 3uided tRNA Adenosine Deaminase in Rice显示文摘 | Fang Yan Yongjie Kuang Bin Ren Jingwen Wang Dawei Zhang Honghui Lin Bing Yang Xueping Zhou Huanbin Zhou | 2018 | Molecular Plant2018，11，4： | 26 |
| 5 | Cas9-NG Greatly Expands the Targeting Scope of the Genome-Editing Toolkit by Recognizing NG and Other Atypical PAMs in Rice显示文摘CRISPR technologies enabling precise genome manipulation are valuable for gene function studies and molecular crop breeding. However, the requirement of a protospacer adjacent motif (PAM)y such as NGG and TTN, for Cas protein recognition restricts the selection of targetable genomic loci in practical applications of CRISPR technologies. Recently Cas9-NG, which recognizes a minimal NG PAM, was reported to expand the targeting space of genome editing in human cells, but it remains unclear whether this Cas9 variant can be used in plants. In this study, we evaluated the nuclease activity of Cas9-NG toward various NGN PAMs by targeting endogenous genes in transgenic rice. We found that Cas9-NG edits all NGG, NGA, NGT, and NGC sites with impaired activity, while the gene-edited plants were dominated by monoallelic mutations. Cas9-NG-engineered base editors were then developed and used to generate O s B Z R I gainof- function plants that can not be created by other available Cas9-engineered base editors. Moreover, we showed that a Cas9-NG-based transcriptional activator efficiently upregulated the expression of endogenous target genes in rice. In addition, we discovered that Cas9-NG recognizes NAC, NTG, NTT, and NCG apart from NG PAM. Together, these findings demonstrate that Cas9-NG can greatly expand the targeting scope of genome-editing tools, showing great potential for targeted genome editing, base editing, and genome regulation in plants. | Bin Ren Lang Liu Shaofang Li Yongjie Kuang Jingwen Wang Dawei Zhang Xueping Zhou Honghui Lin Huanbin Zhou | 2019 | Molecular Plant2019，12，7： | 18 |
| 6 | Petrogenesis, Tectonic Evolution and Geothermal Implications of Mesozoic Granites in the Huangshadong Geothermal Field, South China显示文摘Mesozoic multi-stage tectono-magmatic events produced widely distributed granitoids in the South China Block. Huangshadong(HSD) is located in south-eastern South China Block, where closely spaced hot springs accompany outcrops of Mesozoic granites. New data on whole-rock geochemistry, zircon U-Pb geochronology, and zircon Lu-Hf isotopes are presented, to study the petrogenesis and tectonic evolution of the granites, and to explore the relationship between granites and geothermal anomalies. Zircon U-Pb isotopes display three periods of granites in the HSD area: Indosinian(ca. 253 Ma, G4) muscovite-bearing monzonitic granite, early Yanshanian(ca. 175–155 Ma, G5 and G3) monzonitic granite and granodiorite, and late Yanshanian(ca. 140 Ma, G1 and G2) biotite monzonitic granite. In petrogenetic type, granites of the three periods are I-type granite. Among them, G1, G2, G3, and G4 are characterized by high fractionation, with high values of SiO2, alkalis, Ga/Al, and Rb/Sr, and depletion in Sr, Ba, Zr, Nb, Ti, REEs, with low(La/Yb)N, Nb/Ta, and Zr/Hf ratios and negative Eu anomalies. In terms of tectonic setting, 253 Ma G4 may be the product of partial melting of the ancient lower crust under post-orogenic extensional tectonics, as the closure of the Paleo-Tethys Ocean resulted in an intracontinental orogeny. At 175 Ma, the subduction of the Pacific Plate became the dominant tectonic system, and low-angle subduction of the Paleo-Pacific Plate facilitated partial melting of the subducted oceanic crust and basement to generate the hornblende-bearing I-type granodiorite. As the dip angle of the subducting plate increased, the continental arc tectonic setting was transformed to back-arc extension, inducing intense partial melting of the lower crust at ca. 158 Ma and resulting in the most frequent granitic magmatic activity in the South China hinterland. When slab foundering occurred at ca. 140 Ma, underplating of mantle-derived magmas caused melting of the continental crust, generating extensive highly fractionated granites in HSD. Combining the granitic evolution of HSD and adjacent areas and radioactive heat production rates, it is suggested that highly fractionated granites are connected to the enrichments in U and Th with magma evolution. The high radioactive heat derived from the Yanshanian granites is an important part of the crustal heat, which contributes significantly to the terrestrial heat flow. Drilling ZK8 reveals deep, ca. 140 Ma granite, which implies the heat source of the geothermal anomalies is mainly the concealed Yanshanian granites, combining the granite distribution on the surface. | Zhicai Xiao Shuai Wang Shihua Qi Jian Kuang Min Zhang Feng Tian Yongjie Han | 2020 | Journal of Earth Science2020，31，1： | 9 |
| 7 | Upwelling of Mantle-derived Material in Southeast China:Evidence from Noble Gas Isotopes显示文摘Shallow groundwater collected in Chaozhou,Huizhou,and Guangzhou allowed testing of concentrations and the isotope ratios of noble gases.Based on the calculated noble gas temperature(NGT)and the ratio of noble gas isotopes,the recharge temperature,recharge source,and residence time of groundwater can be calculated.In addition,the contribution of noble gas components from different sources to the sample components can be assessed.In the Huizhou area,according to the 1/Xe vs.Ne/Xe and NGT data,the shallow sandstone-confined water samples in the Shiba area and the unconfined water samples of the Huangshadong are in different temperature ranges,indicating that they have different recharge sources,both in time or space.The He components in the samples are calculated to obtain the content of radiogenic ^(4)He in the crust and to simulate the groundwater ages.The noble gas isotope ratios show the addition of mantle components into the basalt aquifers and sandstone aquifers in Chaozhou and Huizhou.Except for atmospheric and crustal sources,there is a certain proportion of mantle-derived components in the shallow underground cold water in Huizhou and Chaozhou.The noble gases in the Chaozhou groundwater have an obvious mantle signature,allowing speculation that there is a deep fluid carrying mantle characteristics.This upwelling of mantle-derived material might be caused by the India-Eurasia collision or that between the Philippine Sea Plate and the Eurasian Plate. | WANG Shuai KUANG Jian HUANG Xuelian ZHANG Hongyan ZHANG Min QI Shihua HAN Yongjie XIAO Zhicai WANG Siqi TANG Ling | 2022 | Acta Geologica Sinica(English Edition)2022，96，1： | 2 |
| 8 | CRISPR/Sc^(++)-mediated genome editing in rice显示文摘Streptococcus canis Cas9(ScCas9)is an RNAguided endonuclease with NNG protospacer adjacent motif(PAM)specificity whose genomeediting activity in rice is locus-dependent.Here we investigated the performance of a ScCas9 variant named Sc^(++)at different NNG PAM sites in the rice genome;Sc^(++)harbors a T1227K mutation and the substitution of a positively charged loop(residues 367-376).Sc^(++)nuclease achieved broader genome editing compared to the original ScCas9,and its nickase improved targeted base editing in transgenic rice plants.Using the high-efficiency adenine base editor rBE73b,we generated many new OsGS1 alleles suitable for screening of rice germplasm for potential herbicide resistance in the future.The CRISPR/Sc^(++)system expands the genomeediting toolkit for rice. | Guigen Ma Yongjie Kuang Zhenwan Lu Xueqi Li Ziyan Xu Bin Ren Xueping Zhou Huanbin Zhou | 2021 | Journal of Integrative Plant Biology2021，63，9： | 0 |
| 9 | Three novel alleles of OsGS1 developed by base-editing-mediated artificial evolution confer glufosinate tolerance in rice显示文摘Only few glufosinate-tolerant genes,such as phosphinothricin acetyltransferase(PAT)and bialaphos resistance(bar)identified from Streptomyces,are currently available for developing genetically modified rice in agricultural application.Following the rapid development of genome editing technology,generation of novel glufosinate-tolerant gene resources through artificial evolution of endogenous genes is more promising and highly desirable in rice molecular breeding program.In this study,the endogenous Glutamine synthetase1(OsGS1)was artificially evolved by base-editing-mediated gene evolution(BEMGE)in rice cells to create novel alleles conferring glufosinate tolerance in rice germplasms.Two novel glufosinate-tolerant OsGS1 alleles(OsGS1-AVPS and OsGS1-+AF)and one reported tolerant allele(OsGS1-SGTA)were successfully identified from approximately 4200 independent hygromycin-tolerant calli.Germination assays and spray tests revealed that these three OsGS1 alleles confer glufosinate tolerance in rice.Furthermore,OsGS1-AVPS and OsGS1-SGTA were quickly deployed into the elite rice cultivar Nangeng 46 through precise base editing.Overall,our results demonstrate the feasibility of developing glufosinate-tolerant rice by editing an endogenous rice gene in molecular breeding programs. | Bin Ren Yongjie Kuang Ziyan Xu Xuemei Wu Dawei Zhang Fang Yan Xiangju Li Xueping Zhou Guirong Wang Huanbin Zhou | 2023 | The Crop Journal2023，11，2： | 0 |
| 10 | Characteristics of Hydrogen and Oxygen Isotopes and Noble Gas Isotopes in the Groundwater of Weishan, Wudalianchi, Northeast China显示文摘According to the hydrochemical characteristics, hydrogen and oxygen isotope characteristics and the ratio of noble gas isotopes of the sandstone aquifer and basalt aquifer, this study calculated the recharge temperature and residence time of groundwater in the Weishan area of Wudalianchi, also calculating the contribution of noble gas components from different sources to the samples. Based on the characteristics of hydrogen and oxygen isotopes and noble gases Xe and Ne, the recharge altitude and recharge temperature of the two aquifers were estimated, and the recharge temperature fitting with the NGT model as verified, the results showing that the main recharge altitude of groundwater in the region was 500–600 m, the recharge temperature being 2–7°C. He_(eq) and He_(ea) of the samples have been simulated using the OD model, the content of radioactive ~4He in the crust being obtained, the groundwater ages under the two conditions(closed condition and open condition) both being simulated. The results show that groundwater from the sandstone layer water is older than groundwater from the basalt layer. Hydrochemical characteristics and noble gas isotope ratios indicate that in the basalt aquifer and sandstone aquifer in the Weishan area, in addition to atmospheric and crustal helium, there is also an input of mantle-derived helium. The fault constitutes the uplift channel for groundwater containings mantle components, which results in the mantle source composition in water samples near the fault being much higher than those form non-fault areas. | WANG Shuai HUANG Xuelian QI Shihua HAN Yongjie KUANG Jian WANG Siqi WANG He XIAO Zhicai | 2022 | Acta Geologica Sinica(English Edition)2022，96，5： | 0 |
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