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Development and Application of Prime Editing in Plants

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Rice science 2023年第6期30卷 509-522页

作者： LIU Tingting ZOU Jinpeng YANG Xi wang kejian RAO Yuchun wang Chun State Key Laboratory of Rice Biology and Breeding China National Rice Research InstituteHangzhou 310006China College of Life Science and Technology Huazhong Agricultural UniversityWuhan 430070China College of Life Sciences Zhejiang Normal UniversityJinhua 321004China

Clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)-mediated genome editing has greatly accelerated progress in plant genetic research and agricultural breeding by enabling targeted genomic modifications.Moreover,the prime editing system,derived from the CRISPR/Cas system,has opened the door for even more precise genome editing.Prime editing has the capability to facilitate all 12 types of base-to-base conversions,as well as desired insertions or deletions of fragments,without inducing double-strand breaks and requiring donor DNA templet.In a short time,prime editing has been rapidly verified as functional in various plants,and can be used in plant genome functional analysis as well as precision breeding of crops.In this review,we summarize the emergence and development of prime editing,highlight recent advances in improving its efficiency in plants,introduce the current applications of prime editing in plants,and look forward to future prospects for utilizing prime editing in genetic improvement and precision molecular breeding.

关键词： prime editing CRISPR/Cas precision genome editing crop breeding

解锁作物多样性:通过基因组编辑增加遗传变异

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Science Bulletin 2024年第3期69卷 281-284页

作者：邹金鹏黄勇高彩霞王克剑 State Key Laboratory of Rice Biology and Breeding China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China New Cornerstone Science Laboratory Center for Genome EditingInstitute of Genetics and Developmental BiologyChinese Academy of SciencesBeijing 100101China College of Advanced Agricultural Sciences University of Chinese Academy of SciencesBeijing 100049China Hainan Yazhou Bay Seed Laboratory Sanya 572025China

The germplasm resource repository harbors an extensive collection of genetic variations,providing a crucial foundation for the survival and sustainable development of humankind.Throughout history,major agricultural breakthroughs have relied on safeguarding,exploring,and harnessing germplasm resources.However,the pursuit of high yields in modern agriculture has led to a continuous reduction in biodiversity,resulting in monocultures and an undesirable homogeneity of breeding materials.As a consequence,germplasm resources are facing the alarming prospect of accelerated loss leading to a decline in crop diversity.Furthermore,modern agricultural varieties encounter formidable challenges in terms of adapting to unfavorable growing conditions,such as environmental heterogeneity and the prevalence of pests and pathogens(Fig.1a).Enhancing the genetic variability of modern crops becomes paramount for fostering innovation within germplasm resources and ensuring food security.

关键词： germplasm breeding crops

Development and Application of CRISPR/Cas System in Rice

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Rice science 2019年第2期26卷 69-76,I0004-I0006页

作者： REN Jun HU Xixun wang kejian wang Chun State Key Laboratory of Rice Biology China National Rice Research Institute Chinese Academy of Agricultural Sciences

In the past several years, the CRISPR(clustered regularly interspaced short palindromic repeats)/Cas(CRISPR-associated protein) system has been harnessed as an efficient and powerful tool for targeted genome editing in different prokaryotic and eukaryotic species. Here, we review the development and application of CRISPR/Cas system in rice, emphasizing different varieties of CRISPR/Cas systems have been applied, strategies for multiplex editing, methods for precise gene insertion and replacement, and efficient systems for base editing and site-specific transcriptional regulation. In addition, the biosecurity of CRISPR/Cas system is also discussed, including transgene-free methods and off-target effects of CRISPR/Cas system. Thus, the development and application of CRISPR/Cas system will have significant impact on functional genomic research and variety improvement in rice.

关键词： rice CRISPR Cas genome editing protospacer adjacent motif

Regulation of metallothionein gene expression in response to benzo[a]pyrene exposure and bacterial challenge in marine cultured black porgy(Acanthopagrus schlegelii)

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Chinese Journal Of Geochemistry 2014年第4期33卷 404-410页

作者： YANG Ming PAN Chenyuan BO Jun wang kejian School of Environmental and Chemical Engineering Shanghai University State Key Laboratory of Marine Environmental Science Xiamen University

Metallothioneins(MTs) are normally considered to be sensitive indicators of heavy metal pollution, but it is less clear whether the MT gene can actively respond to other environmental stresses. In this study, an MT m RNA molecular sequence of 471 bp(full length) was identified in marine cultured black porgy(Acanthopagrus schlegelii), encoding 60 amino acids containing 20 cysteine residues. The MT sequence was highly homologous to that of other fish belonging to the MT superfamily type 1 family. The three dimensional structure of the deduced MT peptide was composed of two metal-binding domains capable of ligating divalent heavy metals. The MT m RNA transcripts were detected in the 11 tested tissues and the highest quantity was present in the liver. Stresses by two factors, benzo[a]pyrene(B[a]p) exposure and bacterial challenge, were evaluated on MT gene expression. The level of MT gene transcripts in the liver significantly declined 24 h post B[a]p exposure and the quantity was significantly correlated with the exposure time during a 24 h period. In contrast, MT gene expression in the liver was significantly increased 48 h post bacterial infection and the quantity was significantly correlated with the infection time during this period of 48 h. Our results indicated that MT gene expression in black porgy liver was sensitive to environmental stresses other than just the heavy metal pollution reported, suggesting that the development of a reliable biomarker for heavy metal pollution will be more complex than expected.

关键词：金属硫蛋白基因苯并[a]芘细菌感染海洋养殖曝光量黑鲷重金属污染 mRNA转录

Increasing the Genetic Recombination Frequency by Partial Loss of Function of the Synaptonemal Complex in Rice

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Molecular Plant 2015年第8期8卷 1295-1298页

作者： wang, kejian wang, Chun Liu, Qing Liu, Wenzhen Fu, Yaping Chinese Acad Agr Sci China Natl Rice Res Inst State Key Lab Rice Biol Hangzhou 310006 Zhejiang Peoples R China

Dear Editor Genetic recombination results in novel allele combinations and is the most important genetic phenomenon for generating variation in breeding populations. However, in most organisms, the number of genetic recombinations is tightly constrained, which hampers the introgression of new genetic traits and limits genetic variation during agricultural breeding （Barakate et al., 2014）. Therefore, the possibility of increasing genetic recombination frequency has attracted much interest from crop breeders because of the obvious practical applications in traditional breeding （Leflon et al., 2010; Crismani et al., 2012）. However, to date, the goal of increasing the genetic recombination frequency has not been achieved in crop plants.

关键词： MEIOTIC CROSSOVERS MEIOSIS

Biodegradation and Mechanical Property of Polylactic Acid/Thermoplastic Starch Blends with Poly (ethylene glycol)

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Journal of Wuhan University of Technology(Materials Science) 2013年第1期28卷 157-162页

作者：薛平 wang kejian 贾明印 YANG Meijuan Institute of Plastics Machinery and Engineering Beijing University of Chemical Technology

The effects of adding poly （ethylene glycol）（PEG） into polylactic acid/thermoplastic starch blends （PLA/TPS） on the properties were investigated by DSC, SEM and mechanical property-testing. The blends of PLA/TPS blended with increasing content PEG exhibited lower temperature of glass transition （T） and lower temperature of melting （T） as well as higher melt flow index （MFI）, which indicates the plasticization and proeessability of the composites were dramatically improved. The tensile strength, flexural strength and izod impact strength of PLA/TPS （80/20） increased at first and then decreased with increasing content of PEG due to stronger interfacial adhesion. The optimized mechanical property can be obtained for the blend with 3 wt % PEG. The samples containing PEG after soil burial for 5 months showed quicker degradation being accompanied with large weight loss and mechanical properties loss.

关键词： biodegradation mechanical property polylactic acid starch

基于CRV-YOLO的苹果中心花和边花识别方法

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农业机械学报 2024年第2期55卷 278-286页

作者：司永胜孔德浩王克俭刘丽星杨欣河北农业大学信息科学与技术学院保定071001 河北农业大学机电工程学院保定071001

苹果树疏花是果园生产管理中的重要环节。准确高效地识别苹果中心花和边花,是研发智能疏花机器人的前提。针对苹果疏花作业中的实际需求,提出了一种基于CRV-YOLO的苹果中心花和边花识别方法。本文基于YOLO v5s模型进行了如下改进:将C-Co... 详细信息

苹果树疏花是果园生产管理中的重要环节。准确高效地识别苹果中心花和边花,是研发智能疏花机器人的前提。针对苹果疏花作业中的实际需求,提出了一种基于CRV-YOLO的苹果中心花和边花识别方法。本文基于YOLO v5s模型进行了如下改进:将C-CoTCSP结构融入Backbone,更好地学习上下文信息并提高了模型特征提取能力,提高了模型对外形相似和位置关系不明显的中心花和边花的检测性能。在Backbone中添加改进RFB结构,扩大特征提取感受野并对分支贡献度进行加权,更好地利用了不同尺度特征。采用VariFocal Loss损失函数,提高了模型对遮挡等场景下难识别样本检测能力。在3个品种1837幅图像数据集上进行了实验,结果表明,CRV-YOLO的精确率、召回率和平均精度均值分别为95.6%、92.9%和96.9%,与原模型相比,分别提高3.7、4.3、3.9个百分点,模型受光照变化和苹果品种影响较小。与Faster R-CNN、SSD、YOLOX、YOLO v7模型相比,CRV-YOLO的精确率、平均精度均值、模型内存占用量和复杂度性能最优,召回率接近最优。研究成果可为苹果智能疏花提供技术支持。

关键词：苹果花识别 YOLO v5s 上下文信息中心花边花

基于改进ConvNeXt的奶牛行为识别方法

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农业机械学报 2024年第5期55卷 282-289,404页

作者：李恩泽王克俭司永胜苑迎春何振学河北农业大学信息科学与技术学院保定071001 河北农业大学河北省农业大数据重点实验室保定071001

奶牛的动作行为(进食、躺卧、站立、行走和甩尾)直接或间接地反映了奶牛的健康及生理状况,是奶牛疾病监测及感知奶牛异常的关键,为准确高效地对奶牛行为进行识别,提出了一种融合时间和空间注意信息的多分支并行的CAFNet(ConvNeXt-ACM-F... 详细信息

奶牛的动作行为(进食、躺卧、站立、行走和甩尾)直接或间接地反映了奶牛的健康及生理状况,是奶牛疾病监测及感知奶牛异常的关键,为准确高效地对奶牛行为进行识别,提出了一种融合时间和空间注意信息的多分支并行的CAFNet(ConvNeXt-ACM-FAM)奶牛行为识别模型,该模型在卷积网络ConvNeXt的基础上融合非对称多分支卷积模块(ACM)和特征注意力模块(FAM)。首先,利用ACM划分通道分支提取特征并保留一部分原始特征,防止信息过度丢失。其次,FAM对不同通道的特征进行融合并引入SimAM注意力机制,不增加网络参数的同时增强重要特征的有效提取。实验结果表明,该方法对进食、躺卧、站立、行走和甩尾行为识别准确率分别为95.50%、93.72%、90.26%、86.43%、89.39%,平均准确率为91.06%,参数量相较于原模型减少了1.5×10^(6),浮点运算量减少了3×10^(8),相较于其他模型,本文模型识别平均准确率平均提升8.63个百分点。本文研究成果可为奶牛疾病监测及预防提供技术支持。

关键词：奶牛行为识别非对称卷积 SimAM注意力 ConvNeXt

The rice codebook: From reading to editing

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Molecular Plant 2022年第4期15卷 569-572页

作者： kejian wang Huanbin Zhou Qian Qian State Key Laboratory of Rice Biology China National Rice Research InstituteChinese Academy of Agricultural SciencesHangzhou 310006China State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China

Rice(Oryza sativa)is the main staple food crop that feeds more than half of the world's population.Given its relatively small genome size,abundant genetic resources,and highly efficient transformation technology,rice also remains as an excellent model system for monocot crops.In 2002,the International Rice Genome Sequencing Project(IRGSP)released the draft genome sequence of rice variety Nipponbare,the first crop genetic codebook deciphered right after the human genome(Yu et al.,2002).Since then,rice genome annotation,large-scale genome resequencing of diverse rice germplasm accessions,a comprehensive map of rice genome variations,and high-quality reference genome sequence,especially the gap-free reference genome,were completed successively in the next two decades along with the development of high-throughput DNA sequencing techniques(wang et al.,2018;Li et al.,2021;Song et al.,2021).Consequently,rice functional genomics research has boomed rapidly,greatly reshaping our knowledge of plant molecular biology and designed crop breeding.

关键词： sativa germplasm successively