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Sorption kinetics of parent and substituted PAHs for low-density polyethylene（LDPE）:Determining their partition coefficients between LDPE and water(KLDPE) for passive sampling

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Journal of Environmental Sciences 2020年第1期32卷 349-360页

作者： Pei Lei jinjie zhu Ke Pan Hong Zhang State Key Laboratory of Pollution Control and Resources Reuse School of the EnvironmentNanjing UniversityNanjing 210023China State Key Laboratory cf Environmental Aquatic Chemistry Research Center for Eco-Environmental SciencesChinese Academy of SciencesBeijing 100085China Institute for Advanced Study Shenzhen UniversityShenzhen 518060China University of Chinese Academy of Sciences Beijing 100049China

Low-density polyethylene(LDPE)has been widely used as a sorbent for passive sampling of hydrophobic organic contaminants(HOCs)in aquatic ***,it has seen only limited application in passive sampling for measurement of freely dissolved concentrations of parent and substituted PAHs(SPAHs),which are known to be toxic,mutagenic and ***,the 16 priority PAHs and some typical PAHs were selected as target compounds and were simultaneously determined by gas chromatography–mass spectrometer(GC–MS).Some batch experiments were conducted in the laboratory to explore the adsorption kinetics of the target compounds in LDPE *** results showed that both PAHs and SPAHs could reach equilibrium status within19–38 days in sorption kinetic *** coefficients of partitioning between LDPE film(50μm thickness)and water(KLDPE)for the 16 priority PAHs were in good agreement with previously reported values,and the values of KLDPEfor the 9 SPAHs are reported in this study for the first *** linear relationships were observed,i.e.,log KLDPE=0.705×log KOW+1.534 for PAHs(R^2=0.8361,p<0.001)and log KLDPE=0.458×log KOW+3.092 for SPAHs(R^2=0.5609,p=0.0077).The selected LDPE film was also proven to meet the condition of"zero sink"for the selected target *** results could provide basic support for the configuration and in situ application of passive samplers.

关键词： Low-density polyethylene(LDPE) PAHs Substituted PAHs(SPAHs) Freely dissolved concentrations Passive sampling

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Photoelectrochemical glucose biosensor incorporating CdS nanoparticles

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Particuology 2009年第5期7卷 347-352页

作者： jinjie Sun Yihua zhu Xiaoling Yang Chunzhong Li Key Laboratory for Ultrafine Materials of Ministry of Education School of Materials Sdence and EngineeringEast China University of Science and Technology 130 Meilong Road. Shanghai 200237 China

A novel photoelectrochemical biosensor incorporating nanosized CdS semiconductor crystals with enzyme to enhance photochemical reaction has been investigated. CdS nanoparticles were synthesized by using dendrimer PAMAM as inner templates. The CdS nanoparticles and glucose oxidase （GOD） were immobilized on Pt electrode via layer-by-layer （LbL） technique to fabricate a biological-inorganic hybrid system. Under ultraviolet light, the photo-effect of the CdS nanoparticles showed enhancement of the biosensor to detect glucose. Pt nanoparticles were mixed into the Nation film to immobilize the CdS/enzyme composites and to improve the charge transfer of the hybrid. Experimental results demonstrate the desirable characteristics of this biosensing system, e,g. a sensitivity of 1.83 μA/（mM cm^2）, lower detection limit （1 μM）, and acceptable reproducibility and stability,

关键词： CdS nanoparticles PAMAM Photoelectrochemical Biosensor Glucose oxidase Layer-by-layer

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Establishment of an efficient seed fluorescence reporter-assisted CRISPR/Cas9 gene editing in maize

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Journal of Integrative Plant Biology 2021年第9期63卷 1671-1680页

作者： Yuanyuan Yan jinjie zhu Xiantao Qi Beijiu Cheng Changlin Liu Chuanxiao Xie Institute of Crop Science Chinese Academy of Agricultural SciencesNational Key Facility for Crop Gene Resources and Genetic ImprovementBeijing 100081China School of Life Sciences Anhui Agricultural UniversityHefei 230036China

Genome editing by clustered regularly interspaced short palindromic sequences(CRISPR)/CRISPRassociated protein 9(Cas9)has revolutionized functional gene analysis and genetic *** reporter-assisted CRISPR/Cas systems can greatly facilitate the selection of genome-edited plants produced via stable transformation,this approach has not been well established in seed ***,we established the seed fluorescence reporter(SFR)-assisted CRISPR/Cas9 systems in maize(Zea mays L.),using the red fluorescent Ds RED protein expressed in the endosperm(En-SFR/Cas9),embryos(Em-SFR/Cas9),or both tissues(Em/En-SFR/Cas9).All three SFRs showed distinct fluorescent patterns in the seed endosperm and embryo that allowed the selection of seeds carrying the transgene of having segregated the transgene *** describe several case studies of the implementation of En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/Cas9 to identify plants not harboring the genomeediting cassette but carrying the desired mutations at target genes in single genes or in small-scale mutant libraries,and report on the successful generation of single-target mutants and/or mutant libraries with En-SFR/Cas9,Em-SFR/Cas9,and Em/En-SFR/***-assisted genome editing may have particular value for application scenarios with a low transformation frequency and may be extended to other important monocot seed crops.

关键词： CRISPR/Cas9 embryo-specific reporter endosperm-specific reporter genome editing maize seed fluorescence reporter

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Genome Editing Enables Next-Generation Hybrid Seed Production Technology

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Molecular Plant 2020年第9期13卷 1262-1269页

作者： Xiantao Qi Congsheng Zhang jinjie zhu Changlin Liu Changling Huang Xinhai Li Chuanxiao Xie nstitute of Crop Science Chinese Academy of Agricultural SciencesNational Key Facility for Crop Gene Resources and Genetic ImprovementBeijing 100081China Institute of Biotechnology Institute Chinese Academy of Agricultural SciencesBeijing 100081China

The next-generation hybrid seed technology enables the successful production of sortable hybrid seeds from genic male sterile(GMS)lines and maintainers;however,it requires multiple laborious and complicated ***,we designed a simple next-generation hybrid seed production strategy that takes advantage of the CRISPR/Cas9 technology to create a Manipulated GMS Maintainer(MGM)system via a single *** this schema,the maize male fertility gene ZmMS26 was nullified by removal of its fifth exon using the CRISPR/Cas9 system on a vector,and a second vector carrying a functional ZmMS26 cDNA was co-transformed to restore *** second vector also contains a male gametophyte inactivation gene(ZmAA1)encoding maizeα-amylase driven by the pollen-specific promoter PG47 and an endosperm fluorescent marker(DsRED)driven by the barley endosperm aleurone-specific promoter *** derived single-copy hemizygous MGM lines bore a mutated MS26 gene,leading to complete male sterility but normal vegetative growth and grain *** MGM system could prevent genetic transmission of the MGM elements via male gametophytes,providing an efficient method for sorting maintainer seeds labeled by *** strategy can be extended to any GMS gene and to hybrid crops other than maize.

关键词： CRISPR/Cas9 genic male sterility manipulated maintainer ZmMS26 exon deletion

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Efficiency and Inheritance of Targeted Mutagenesis in Maize Using CRISPR-Cas9

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Journal of Genetics and Genomics 2016年第1期43卷 25-36页

作者： jinjie zhu Ning Song Silong Sun Weilong Yang Haiming Zhao Weibin Song Jinsheng Lai State Key Laboratory of Agrobiotechnology and National Maize Improvement Center Department of Plant Genetics and Breeding China Agricultural University

CRISPR/Cas （clustered regularly interspaced short palindromic repeats/CR1SPR-associated proteins） is an adaptive immune system in bacteria and archaea to defend against invasion from foreign DNA fragments. Recently, it has been developed as a powerful targeted genome editing tool for a wide variety of species. However, its application in maize has only been tested with transiently expressed somatic cells or with a limited number of stable transgenic To plants. The exact efficiency and specificity of the CRISPR/Cas system in the highly complex maize genome has not been documented yet. Here we report an extensive study of the well-studied type II CRISPR-Cas9 system for targeted genome editing in maize, with the codon-optimized Cas9 protein and the short non-coding guide RNA generated through a functional maize U6 snRNA promoter. Targeted gene mutagenesis was detected for 90 loci by maize protoplast assay, with an average cleavage efficiency of 10.67 %. Stable knockout transformants for maize phytoene synthase gene （PSYI） were obtained. Mutations occurred in germ ceils can be stably inherited to the next generation. Moreover, no off-target effect was detected at the computationally predicted putative off-target loci. No significant difference between the transcriptomes of the Cas9 expressed and non-expressed lines was detected. Our results confirmed that the CRISPR-Cas9 could be successfully applied as a robust targeted genome editing system in maize.

关键词： CRISPR-Cas9 Targeted mutagenesis Heritability Maize

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A field-deployable method for single and multiplex detection of DNA or RNA from pathogens using Cas12 and Cas13

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Science China(Life Sciences) 2022年第7期65卷 1456-1465页

作者： Lina Li Canxing Duan Jianfeng Weng Xiantao Qi Changlin Liu Xinhai Li jinjie zhu Chuanxiao Xie National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop ScienceChinese Academy of Agricultural Sciences(CAAS)Beijing 100081 China Biotechnology Research Institute Chinese Academy of Agricultural Sciences(CAAS)Beijing 100081China

For some Cas nucleases,trans-cleavage activity triggered by CRISPR/Cas-mediated cis-cleavage upon target nucleic acid recognition has been explored for diagnostic *** single and multiplex nucleic acid-based detection is needed for crop pathogen management in ***,we harnessed and characterized RfxCas13d as an additional CRISPR/Cas nucleic acid detection *** systematically characterized AsCas12a,LbCas12a,LwaCas13a,and RfxCas13d combined with isothermal amplification to develop a CRISPR/Cas nucleic acid-based tool for single or multiplex pathogen *** data indicated that sufficient detection sensitivity was achieved with just a few copies of DNA/RNA targets as *** this tool,we successfully detected DNA from Fusarium graminearum and Fusarium verticillioides and RNA from rice black-streaked dwarf virus in crude extracts prepared in the *** method,from sample preparation to result readout,could be rapidly and easily deployed in the *** system could be extended to other crop pathogens,including those that currently lack a detection method and have metabolite profiles that make detection *** nucleic acid detection system could also be used for single-nucleotide polymorphism genotyping,transgene detection,and qualitative detection of gene expression in the field.

关键词： nucleic acid detection AsCas12a LbCas12a LwaCas13a RfxCas13d maize ear rot Fusarium head blight rice black-streaked dwarf virus(RBSDV)

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Precise base editing of non-allelic acetolactate synthase genes confers sulfonylurea herbicide resistance in maize

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The Crop Journal 2020年第3期8卷 449-456页

作者： Yanmin Li jinjie zhu Hao Wu Changlin Liu Changling Huang Jinhao Lan Yanming Zhao Chuanxiao Xie Institute of Crop Sciences Chinese Academy of Agricultural SciencesNational Key Facility for Crop Gene Resources and Genetic ImprovementBeijing 100081China Qingdao Agricultural University Qingdao 266109ShandongChina Anhui Agricultural University Hefei 230036AnhuiChina

Single-nucleotide polymorphisms contribute to phenotypic diversity in maize. Creation and functional annotation of point mutations has been limited by the low efficiency of conventional methods based on random mutation. An efficient tool for generating targeted single-base mutations is desirable for both functional genomics and precise genetic improvement. The objective of this study was to test the efficiency of targeted C-to-T base editing of two non-allelic acetolactate synthase(ALS) in generating sulfonylurea herbicide-resistant mutants. A CRISPR/Cas9 nickase-cytidine deaminase fused with uracil DNA glycosylase inhibitor(UGI) was employed to achieve targeted conversion of cytosine to thymine in ZmALS1 and ZmALS2. Both protoplasts and recovered mutant plants showed the activity of the cytosine base editor, with an in vivo efficiency of up to 13.8%. Transgene-free edited plants harboring a homozygous ZmALS1 mutation or a ZmALS1 and ZmALS2 double mutation were tested for their resistance at a dose of up to 15-fold the recommended limit of chlorsulfuron, a sulfonylurea herbicide widely used in agriculture. Targeted base editing of C-to-T per se and a phenotype verified in the generated mutants demonstrates the power of base editing in precise maize breeding.

关键词： Precise base editing of non-allelic acetolactate synthase genes confers sulfonylurea herbicide resistance in maize

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Conversion of a normal maize hybrid into a waxy version using in vivo CRISPR/Cas9 targeted mutation activity

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The Crop Journal 2020年第3期8卷 440-448页

作者： Xiantao Qi Hao Wu Haiyang Jiang jinjie zhu Changling Huang Xin Zhang Changlin Liu Beijiu Cheng Anhui Agricultural University Hefei 230036AnhuiChina Institute of Crop Sciences Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic ImprovementBeijing 100081China

Waxy maize is a specialty maize that produces mainly amylopectin starch with special food or industrial values. The objective of this study was to overcome the limitations of wx mutant allele acquisition and breeding efficiency by conversion of parental lines from normal to waxy maize. The intended mutation activity was achieved by in vivo CRISPR/Cas9 machinery involving desired-target mutation of the Wx locus in the ZC01 background,abbreviated as ZC01-DTM^(wx). Triple selection was applied to segregants to obtain high genome background recovery with transgene-free wx mutations. The targeted mutation was identified, yielding six types of mutations among progeny crossed with ZC01-DTM^(wx).The amylopectin contents of the endosperm starch in mutant lines and hybrids averaged94.9%, while those of the wild-type controls were significantly(P < 0.01) lower, with an average of 76.9%. Double selection in transgene-free lines was applied using the Bar strip test and Cas9 PCR screening. The genome background recovery ratios of the lines were determined using genome-wide SNP data. That of lines used as male parents was as high as98.19% and that of lines used as female parents was as high as 86.78%. Conversion hybrids and both parental lines showed agronomic performance similar to that of their wild-type counterparts. This study provides a practical example of the efficient extension of CRISPR/Cas9 targeted mutation to industrial hybrids for transformation of a recalcitrant species.

关键词： Conversion of a normal maize hybrid into a waxy version using in vivo CRISPR/Cas9 targeted mutation activity CRISPR

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Engineering high amylose and resistant starch in maize by CRISPR/Cas9-mediated editing of starch branching enzymes

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The Crop Journal 2024年第4期12卷 1252-1258页

作者： Mingzheng Ma Shanqiu Sun jinjie zhu Xiantao Qi Gaoke Li Jianguang Hu Chuanxiao Xie Changlin Liu State Key Laboratory of Crop Gene Resources and Breeding Institute of Crop SciencesChinese Academy of Agricultural SciencesBeijing 100081China Hainan Yazhou Bay Seed Lab Sanya 572024HainanChina Anhui Agricultural University Hefei 230036AnhuiChina Crops Research Institute Guangdong Academy of Agricultural SciencesGuangzhou 510640GuangdongChina

To improve the amylose content(AC)and resistant starch content(RSC)of maize kernel starch,we employed the CRISPR/Cas9 system to create mutants of starch branching enzyme I(SBEI)and starch branching enzyme IIb(SBEIIb).A frameshift mutation in SBEI(E1,a nucleotide insertion in exon 6)led to plants with higher RSC(1.07%),lower hundred-kernel weight(HKW,24.71±0.14 g),and lower plant height(PH,218.50±9.42 cm)compared to the wild type(WT).Like the WT,E1 kernel starch had irregular,polygonal shapes with sharp edges.A frameshift mutation in SBEIIb(E2,a four-nucleotide deletion in exon 8)led to higher AC(53.48%)and higher RSC(26.93%)than that for the WT.E2 kernel starch was significantly different from the WT regarding granule morphology,chain length distribution pattern,X-ray diffraction pattern,and thermal characteristics;the starch granules were more irregular in shape and comprised typical B-type *** SBEI and SBEIIb(E12)had a synergistic effect on RSC,HKW,PH,starch properties,and starch biosynthesis-associated gene ***,SS1,SSIIa,SSIIIa,and SSIIIb were upregulated in E12 endosperm compared to WT *** study lays the foundation for rapidly improving the starch properties of elite maize lines.

关键词： Maize Gene editing Starch branching enzyme I Starch branching enzyme IIb

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OS1 functions in the allocation of nutrients between the endosperm and embryo in maize seeds

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Journal of Integrative Plant Biology 2019年第6期61卷 706-727页

作者： Weibin Song jinjie zhu Haiming Zhao Yingnan Li Jiangtao Liu Xiangbo Zhang Liangliang Huang Jinsheng Lai State Key Laboratory of Agrobiotechnology and National Maize Improvement Center Department of Plant Genetics and Breeding ChinaAgricultural University

Uncovering the genetic basis of seed develop-ment will provide useful tools for improving both crop yield and nutritional value, However, the genetic regulatory networks of maize (Zea mays) seed development remain largely unknown. The maize opaque endosperm and small germ 1 (osl) mutant has opaque endosperm and a small embryo,Here, we cloned OSi1 and show that it encodes a putative transcription factor containing an RWP-RK domain-Transcriptional analysis indicated that OSi1 expression is elevated in early endosperm development, especially in the basal endosperm transfer layer (BETL), conducting zone (CZ), and central starch endospcrm (CSE) cells. RNA sequencing (RNA-Seq) analysis of the osi mutant revealed sharp downregulation of certain genes in specific cell types, including ZmMRP-1 and Megl in BETL cells and a majority of zein-and starch-related genes in CSE cells. Using a haploid induction system, we show that wild-type endosperm could rescue the smaller size of osi embryo, which suggests that nutrients are allocated by tho wild type endosperm. Therefore, our dsta imply that the network regulated by 051 accomplishes a key step in nutrient allocation between endosperm and embryo within maize seeds. Identification of this network will help uncover the mechanisms regulating the nutritional balance between endosperm and embryo.

关键词： 0S1 functions the allocation of nutrients between the endosperm and embryo in maize seeds^FA

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