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m^(6)A modification of plant virus enables host recognition by NMD factors in plants

Science China(Life Sciences) 2024年第1期67卷 161-174页

作者： Hao He Linhao Ge Yalin Chen Siwen Zhao Zhaolei Li Xueping Zhou Fangfang Li State Key Laboratory for Biology of Plant Diseases and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural SciencesBeijing 100193China State Key Laboratory of Rice Biology Institute of BiotechnologyZhejiang UniversityHangzhou 310029China

N^(6)-methyladenosine(m^(6)A)is the most abundant eukaryotic mRNA modification and is involved in various biological processes.Increasing evidence has implicated that m^(6)Amodification is an important anti-viral defense mechanism in mammals and plants,but it is largely unknown how m^(6)Aregulates viral infection in plants.Here we report the dynamic changes and functional anatomy of m^(6)Ain Nicotiana benthamiana and Solanum lycopersicum during Pepino mosaic virus(PepMV)infection.m^(6)Amodification in the PepMV RNA genome is conserved in these two species.Overexpression of the m^(6)Awriters,mRNA adenosine methylase A(MTA),and HAKAI inhibit the PepMV RNA accumulation accompanied by increased viral m^(6)Amodifications,whereas deficiency of these writers decreases the viral RNA m^(6)Alevels but enhances virus infection.Further study reveals that the cytoplasmic YTH-domain family protein NbECT2A/2B/2C as m^(6)Areaders are involved in anti-viral immunity.Protein-protein interactions indicate that NbECT2A/2B/2C interact with nonsense-mediated mRNA decay(NMD)-related proteins,including NbUPF3 and NbSMG7,but not with NbUPF1.m^(6)Amodification-mediated restriction to PepMV infection is dependent on NMD-related factors.These findings provide new insights into the functionality of m^(6)Aanti-viral activity and reveal a distinct immune response that NMD factors recognize the m^(6)Areaders-viral m^(6)ARNA complex for viral RNA degradation to limit virus infection in plants.

关键词： m^(6)A Pepino mosaic virus plant defense m^(6)A readers NMD factors

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Molecular mechanism of exopolysaccharide biosynthesis regulated by wze and wzd genes on the immunomodulatory effects of probiotic Lacticaseibacillus paracasei S-NB

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Food Science and Human Wellness 2024年

作者： Luyao Xiao Qian Yang Changliang Zhang Kai Ma Xiaogan Zhao Xin Rui Wei Li Jiangsu Biodep biotechnology Co. Ltd. Jiangsu New-Bio biotechnology Co. Ltd. Sanya institute of Nanjing agricultural University College of Food Science and Technology Nanjing agricultural University

The surface macromolecules of probiotic bacteria play crucial roles in modulating immune responses in the host. Exopolysaccharide (EPS) from lactic acid bacteria (LAB) have been widely reported to exhibit immunomodulatory activity. In this study， the EPS biosynthesis gene cluster of Lacticaseibacillus paracasei S-NB was analyzed and a deletion mutant S-NB△7576 (two genes S-NB＿2175/wze and S-NB＿2176/wzd were responsible for the chain length determination and export of EPS) was successfully constructed， resulting a 40.02% decrease in the production of EPS. The deletion of wze and wzd had little effect on the monosaccharide composition and major groups of the two EPS fractions (BEPS1 and BEPS2). Both BEPS1 and BEPS2 could inhibit the transcriptional level of tumor necrosis factor-α (TNF-α)， interleukin-1β (IL-1β)， inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) mRNA in RAW 264.7 cells induced by lipopolysaccharide (LPS)， and enhance host immune tolerance via suppressing NF-κB and MAPK signaling. Notably， the S-NB△7576 mutant supplied with the BEPS1/BEPS2 exhibited more significant inhibition of cytokines production and the phosphorylation of p65 and c-Jun N-terminal kinase (JNK) in LPS-stimulated cells compared with the S-NB△7576 mutant alone. Our study provided the immunomodulatory effect of BEPS1 and BEPS2 from L. paracasei S-NB， in which the wze and wzd genes associated with EPS biosynthesis may play an important role.

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Genetics of biochemical attributes regulating morpho-physiology of upland cotton under high temperature conditions

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Journal of Cotton research 2024年第1期7卷 29-44页

作者： MAJEED Sajid CHAUDHARY Muhammad Tanees MUBARIK Muhammad Salman RANA Iqrar Ahmad SHABAN Muhammad TAN Daniel KY JIA Yinhua DU Xiongming HINZE Lori AZHAR Muhammad Tehseen Federal Seed Certification and Registration Department Ministry of National Food Security and ResearchIslamabad 44090Pakistan Horticultural research institute National Agricultural Research CentreIslamabad 44000Pakistan Department of biotechnology University of NarowalNarowal 51600Pakistan Centre of agricultural Biochemistry and biotechnology University of AgricultureFaisalabad 38400Pakistan Department of Plant Breeding and Genetics University of Agriculture FaisalabadSub-Campus BurewalaVehariPakistan Faculty of Science School of Life and Environ-mental SciencesPlant Breeding InstituteSydney Institute of AgricultureThe University of SydneySydneyNSW 2006Australia State Key Laboratory of Cotton Biology Institute of Cotton Research of Chinese Academy of Agri-cultural ScienceAnyang 455000China agricultural research Service USDASouthern Plains Agricultural Research CentreCollege StationTX 77845USA Department of Plant Breeding and Genetics University of AgricultureFaisal-abad 38400Pakistan School of Agriculture Sciences Zhengzhou UniversityZhengzhou 450000China.

Background Cotton is a strategically important fibre crop for global textile industry.It profoundly impacts several countries’industrial and agricultural sectors.Sustainable cotton production is continuously threatened by the unpre-dictable changes in climate,specifically high temperatures.Breeding heat-tolerant,high-yielding cotton cultivars with wide adaptability to be grown in the regions with rising temperatures is one of the primary objectives of modern cotton breeding programmes.Therefore,the main objective of the current study is to figure out the effective breed-ing approach to imparting heat tolerance as well as the judicious utilization of commercially significant and stress-tolerant attributes in cotton breeding.Initially,the two most notable heat-susceptible(FH-115 and NIAB Kiran)and tolerant(IUB-13 and GH-Mubarak)cotton cultivars were spotted to develop filial and backcross populations to accom-plish the preceding study objectives.The heat tolerant cultivars were screened on the basis of various morphological(seed cotton yield per plant,ginning turnout percentage),physiological(pollen viability,cell membrane thermostabil-ity)and biochemical(peroxidase activity,proline content,hydrogen peroxide content)parameters.Results The results clearly exhibited that heat stress consequently had a detrimental impact on every studied plant trait,as revealed by the ability of crossing and their backcross populations to tolerate high temperatures.However,when considering overall yield,biochemical,and physiological traits,the IUB-13×FH-115 cross went over particularly well at both normal and high temperature conditions.Moreover,overall seed cotton yield per plant exhibited a posi-tive correlation with both pollen viability and antioxidant levels(POD activity and proline content).Conclusions Selection from segregation population and criteria involving pollen viability and antioxidant levels concluded to be an effective strategy for the screening of heat-tolerant cotton germplasms.Therefore,understanding acquired from

关键词： Accessions Biochemical Breeding Cotton Generation mean analysis Heat stress

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A novel semi-dominant allele of the transmembrane NAC transcription factor ZmNTL2 reduces the size of multiple maize organs

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The Crop Journal 2024年第3期12卷 676-687页

作者： Chuan Li Yun Bai Yuwei Hou Siyu Wang Yue Xin Tao Yu Jing Wang Xiaowei Liu Hongyang Yi Chao Xia Hai Lan Moju Cao State Key Laboratory of Crop Gene Exploration and Utilization in Southwest China Maize Research InstituteSichuan Agricultural UniversityChengdu 611130SichuanChina Jilin Engineering research Center for Crop biotechnology Breeding College of Plant ScienceJilin UniversityChangchun 130012JilinChina

Maize plant architecture influences planting density and,in turn,grain yield.Most of the plant architecture-related traits can be described as organ size.We describe a miniature maize mutant,Tiny plant 4(Tip4),which exhibits reduced size of multiple organs and exhibits a semi-dominant monofactorial inheritance characteristic.Positional cloning confirmed that a 4-bp deletion in the NAC TF with transmembrane motif 1-Like(NTL)gene ZmNTL2,denoted as ZmNTL2^(Δ),confers the Tip4 mutation.qRT-PCR showed that ZmNTL2 was expressed in all tested tissues.ZmNTL2 functions as a transcriptional activator and is located in both the nucleus and biomembranes.The mutation does not affect the mRNA abundance of ZmNTL2 locus,but it does result in the loss of transmembrane domain and confines the ZmNTL2^(Δ)protein to the nucleus.Knocking out ZmNTL2 has no effect on maize organ size development,indicating that the 4-bp deletion might be a gain-of-function mutation in organ size regulation.Combining transcriptome sequencing with cytokinin and auxin content determination suggests that the decreased organ size may be possibly mediated by changes in hormone homeostasis.

关键词： Organ size c-ray mutagenesis Membrane-bound NAC TF Hormone homeostasis Maize

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Effects of weaning on intestinal longitudinal muscle-myenteric plexus function in piglets

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Science China(Life Sciences) 2024年第2期67卷 379-390页

作者： Jing Li Fenfen Liu Kaibin Mo Hengjia Ni Yulong Yin Key Laboratory of Agro-Ecological Processes in Subtropical Region Hunan Provincial Key Laboratory of Animal Nutritional Physiology and Metabolic ProcessHunan Research Center of Livestock and Poultry SciencesSouth Central Experimental Station of Animal Nutrition and Feed Science in the Ministry of AgricultureInstitute of Subtropical AgricultureChinese Academy of SciencesChangsha 410125China University of Chinese Academy of Sciences Beijing 100008China Guangdong Key Laboratory for Veterinary Drug Development and Safety Evaluation College of Veterinary MedicineSouth China Agricultural UniversityGuangzhou 510642China

Weaning piglets usually suffer from severe diarrhea(commonly known as postweaning diarrhea,PWD)along with intestinal motility disorder.Intestinal peristalsis is mainly regulated by the longitudinal muscle-myenteric plexus(LM-MP).To understand the relationship between intestinal LM-MP function and the development of PWD,we compared the intestinal electrical activity,and the transcriptional profile of the LM-MP between 21-day-old piglets(just weaned,n=7)and 24-day-old piglets(suffered the most severe weaning stress,n=7).The results showed that 24-day-old piglets exhibited different degrees of diarrhea.A significant increase in the slow-wave frequency in the ileum and colon was observed in 24-day-old piglets,while c-kit expression in the intestinal LM-MPs was significantly decreased,indicating that PWD caused by elevated slow-wave frequency may be associated with loss of c-kit.The real-time quantitative PCR(RT-q PCR)and enzyme-linked immunosorbent assay(ELISA)showed that intestinal LM-MPs in 24-day-old piglets may undergo inflammation and oxidative stress.Significant increases in 8-hydroxy-2′-deoxyguanosine and decreases in thioredoxin suggest that weaning may lead to DNA damage in the LM-MP of 24-day-old piglets.In addition,activating transcription factor 3 was significantly upregulated,indicating nerve damage in the LM-MP of 24-day-old piglets.The transcriptomic results showed that most of the differentially expressed genes in the ileal LM-MP after weaning were downregulated and closely related to the cell cycle process.Subsequent RT-q PCR analysis showed that the relative expression of p21 was upregulated,while the expression of cyclin A2,cyclin B1,and proliferating cell nuclear antigen was downregulated in the ileal and colonic LM-MP of 24-day-old piglets,suggesting that weaning may inhibit cell proliferation and cause G1/S cell cycle arrest in ileal and colonic LM-MP.In conclusion,weaning may lead to cell cycle arrest by causing DNA damage in the LM-MP,impairing intestinal motility regulation,and ultimately lea

关键词： weaning piglets diarrhea longitudinal muscle-myenteric plexus

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Chromatin condensation but not DNA integrity of pig sperm is greater in the sperm-rich fraction

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Journal of Animal Science and biotechnology 2024年第1期15卷 171-181页

作者： Estel Viñolas-Vergés Jordi Ribas-Maynou Isabel Barranco Camila Peres Rubio Sergi Bonet Jordi Roca Marc Yeste Unit of Cell Biology Department of BiologyFaculty of SciencesUniversity of GironaGironaSpain biotechnology of Animal and Human Reproduction(TechnoSperm) Institute of Food and Agricultural TechnologyUniversity of GironaGironaSpain Department of Medicine and Animal Surgery Faculty of Veterinary ScienceUniversity of MurciaMurciaSpain Catalan Institution for research and Advanced Studies(ICREA) S08010 BarcelonaSpain

Background Protamination and condensation of sperm chromatin as well as DNA integrity play an essential role during fertilization and embryo development.In some mammals,like pigs,ejaculates are emitted in three separate fractions:pre-sperm,sperm-rich(SRF)and post sperm-rich(PSRF).These fractions are known to vary in volume,sperm concentration and quality,as well as in the origin and composition of seminal plasma(SP),with differences being also observed within the SRF one.Yet,whether disparities in the DNA integrity and chromatin condensation and pro-tamination of their sperm exist has not been interrogated.Results This study determined chromatin protamination(Chromomycin A3 test,CMA_(3)),condensation(Dibromobi-mane test,DBB),and DNA integrity(Comet assay)in the pig sperm contained in the first 10 m L of the SRF(SRF-P1),the remaining portion of the sperm-rich fraction(SRF-P2),and the post sperm-rich fraction(PSRF).While chromatin protamination was found to be similar between the different ejaculate fractions(P>0.05),chromatin condensation was seen to be greater in SRF-P1 and SRF-P2 than in the PSRF(P=0.018 and P=0.004,respectively).Regarding DNA integrity,no differences between fractions were observed(P>0.05).As the SRF-P1 has the highest sperm concentra-tion and ejaculate fractions are known to differ in antioxidant composition,the oxidative stress index(OSi)in SP,calcu-lated as total oxidant activity divided by total antioxidant capacity,was tested and confirmed to be higher in the SRF-P1 than in SRF-P2 and PSRF(0.42±0.06 vs.0.23±0.09 and 0.08±0.00,respectively;P<0.01);this index,in addition,was observed to be correlated to the sperm concentration of each fraction(Rs=0.973;P<0.001).Conclusion While sperm DNA integrity was not found to differ between ejaculate fractions,SRF-P1 and SRF-P2 were observed to exhibit greater chromatin condensation than the PSRF.This could be related to the OSi of each fraction.

关键词： Chromatin Condensation DNA integrity Ejaculate fractions Pig Protamination Sperm

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IbNIEL-mediated degradation of IbNAC087 regulates jasmonic acid-dependent salt and drought tolerance in sweet potato

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Journal of Integrative Plant Biology 2024年第2期66卷 176-195页

作者： Xu Li Zhen Wang Sifan Sun Zhuoru Dai Jun Zhang Wenbin Wang Kui Peng Wenhao Geng Shuanghong Xia Qingchang Liu Hong Zhai Shaopei Gao Ning Zhao Feng Tian Huan Zhang Shaozhen He Key Laboratory of Sweet Potato Biology and biotechnology Ministry of Agriculture and Rural Affairs/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis&Utilization and Joint Laboratory for International Cooperation in Crop Molecular BreedingMinistry of EducationCollege of Agronomy&BiotechnologyChina Agricultural UniversityBeijing 100193China Sanya institute of China agricultural University Sanya 572025China State Key Laboratory of Plant Physiology and Biochemistry National Maize Improvement CenterKey Laboratory of Biology and Genetic Improvement of MaizeBeijing Key Laboratory of Crop Genetic ImprovementChina Agricultural UniversityBeijing 100193China

Sweet potato(Ipomoea batatas[L.]Lam.)is a crucial staple and bioenergy crop.Its abiotic stress tolerance holds significant importance in fully utilizing marginal lands.Transcriptional processes regulate abiotic stress responses,yet the molecular regulatory mechanisms in sweet potato remain unclear.In this study,a NAC(NAM,ATAF1/2,and CUC2)transcription factor,IbNAC087,was identified,which is commonly upregulated in salt-and drought-tolerant germplasms.Overexpression of IbNAC087 increased salt and drought tolerance by increasing jasmonic acid(JA)accumulation and activating reactive oxygen species(ROS)scavenging,whereas silencing this gene resulted in opposite phenotypes.JA-rich IbNAC087-OE(overexpression)plants exhibited more stomatal closure than wild-type(WT)and IbNAC087-Ri plants under NaCl,polyethylene glycol,and methyl jasmonate treatments.IbNAC087 functions as a nuclear transcriptional activator and directly activates the expression of the key JA biosynthesisrelated genes lipoxygenase(IbLOX)and allene oxide synthase(IbAOS).Moreover,IbNAC087 physically interacted with a RING-type E3 ubiquitin ligase NAC087-INTERACTING E3 LIGASE(IbNIEL),negatively regulating salt and drought tolerance in sweet potato.IbNIEL ubiquitinated IbNAC087 to promote 26S proteasome degradation,which weakened its activation on IbLOX and IbAOS.The findings provide insights into the mechanism underlying the IbNIEL-IbNAC087 module regulation of JA-dependent salt and drought response in sweet potato and provide candidate genes for improving abiotic stress tolerance in crops.

关键词： abiotic tolerance IbNAC087 IbNIEL JA sweet potato

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Generation of double knockout cattle via CRISPR-Cas9 ribonucleoprotein(RNP)electroporation

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Journal of Animal Science and biotechnology 2024年第1期15卷 456-462页

作者： Gyeong-Min Gim Kyeong-Hyeon Eom Dong-Hyeok Kwon Dae-Jin Jung Dae-Hyun Kim Jun-Koo Yi Jae-Jung Ha Ji-Hyun Lee Seong-Beom Lee Woo-Jae Son Soo-Young Yum Won-Wu Lee Goo Jang Laboratory of Theriogenology and biotechnology Department of Veterinary Clinical ScienceCollege of Veterinary Medicine and the Research Institute of Veterinary ScienceSeoul National UniversitySeoulRepublic of Korea LARTBio Co. Ltd.SeoulRepublic of Korea Gyeongsangbukdo Livestock research institute YeongjuRepublic of Korea Comparative Medicine Disease research Center Seoul National UniversitySeoulRepublic of Korea

Background Genome editing has been considered as powerful tool in agricultural fields.However,genome editing progress in cattle has not been fast as in other mammal species,for some disadvantages including long gestational periods,single pregnancy,and high raising cost.Furthermore,technically demanding methods such as microinjection and somatic cell nuclear transfer(SCNT)are needed for gene editing in cattle.In this point of view,electroporation in embryos has been risen as an alternative.Results First,editing efficiency of our electroporation methods were tested for embryos.Presence of mutation on embryo was confirmed by T7E1 assay.With first combination,mutation rates for MSTN and PRNP were 57.6%±13.7%and 54.6%±13.5%,respectively.In case of MSTN/BLG,mutation rates were 83.9%±23.6%for MSTN,84.5%±18.0%for BLG.Afterwards,the double-KO embryos were transferred to surrogates and mutation rate was identified in resultant calves by targeted deep sequencing.Thirteen recipients were transferred for MSTN/PRNP,4 calves were delivered,and one calf underwent an induction for double KO.Ten surrogates were given double-KO embryos for MSTN/BLG,and four of the six calves that were born had mutations in both genes.Conclusions These data demonstrated that production of genome edited cattle via electroporation of RNP could be effectively applied.Finally,MSTN and PRNP from beef cattle and MSTN and BLG from dairy cattle have been born and they will be valuable resources for future precision breeding.

关键词： Beta-lactoglobulin Cattle CRISPR-Cas9 Electroporation Knockout MSTN PRNP

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Two types of coumarins-specific enzymes complete the last missing steps in pyran- and furanocoumarins biosynthesis

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Acta Pharmaceutica Sinica B 2024年第2期14卷 869-880页

作者： Yucheng Zhao Yuedong He Liangliang Han Libo Zhang Yuanzheng Xia Fucheng Yin Xiaobing Wang Deqing Zhao Sheng Xu Fei Qiao Yibei Xiao Lingyi Kong Department of Resources Science of Traditional Chinese Medicines School of Traditional Chinese Pharmacyand State Key Laboratory of Natural MedicinesChina Pharmaceutical UniversityNanjing 210009China College of Bioscience and biotechnology Hunan Agricultural UniversityChangsha 410128China Jiangsu Key Laboratory of Bioactive Natural Product research and State Key Laboratory of Natural Medicines Department of Natural Medicinal ChemistryChina Pharmaceutical UniversityNanjing 210009China Key Laboratory of Crop Gene Resources and Germplasm Enhancement in Southern China Ministry of AgricultureTropical Crops Genetic Resources InstituteChinese Academy of Tropical Agricultural SciencesHaikou 517317China institute of Botany Jiangsu Province and Chinese Academy of SciencesNanjing 210014China Department of Pharmacology School of Pharmacyand State Key Laboratory of Natural MedicinesChina Pharmaceutical UniversityNanjing 210009China

Pyran-and furanocoumarins are key representatives of tetrahydropyrans and tetrahydrofurans,respectively,exhibiting diverse physiological and medical bioactivities.However,the biosynthetic mechanisms for their core structures remain poorly understood.Here we combined multiomics analyses of biosynthetic enzymes in Peucedanum praeruptorum and in vitro functional verification and identified two types of key enzymes critical for pyran and furan ring biosynthesis in plants.These included three distinct P.praeruptorum prenyltransferases(PpPT1e3)responsible for the prenylation of the simple coumarin skeleton 7 into linear or angular precursors,and two novel CYP450 cyclases(PpDC and PpOC)crucial for the cyclization of the linear/angular precursors into either tetrahydropyran or tetrahydrofuran scaffolds.Biochemical analyses of cyclases indicated that acid/base-assisted epoxide ring opening contributed to the enzyme-catalyzed tetrahydropyran and tetrahydrofuran ring refactoring.The possible acid/base-assisted catalytic mechanisms of the identified cyclases were theoretically investigated and assessed using site-specific mutagenesis.We identified two possible acidic amino acids Glu303 in PpDC and Asp301 in PpOC as vital in the catalytic process.This study provides new enzymatic tools in the epoxide formation/epoxide-opening mediated cascade reaction and exemplifies how plants become chemically diverse in terms of enzyme function and catalytic process.

关键词： Tetrahydropyrans Tetrahydrofurans Baldwin rules Coumarins CYP450cyclase

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Multiple pollutants from crop and livestock production in the Yangtze River: status and challenges

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Frontiers of agricultural Science and Engineering 2024年第1期11卷 134-139,I0005页

作者： Lihua MA Shiyang LI Linfa FANG Xuanjing CHEN Ran XIAO Xiaoxuan SU Zhaolei LI Zhaohai BAI Lin MA Prakash LAKSHMANAN Xinping CHEN Interdisciplinary research Center for Agriculture Green Development in Yangtze River Basin College of Resources and EnvironmentSouthwest UniversityChongqing 400715China Water Systems and Global Change Group Wageningen University&Research6708 PB Wageningenthe Netherlands Key Laboratory of agricultural Water Resources Center for Agricultural Resources ResearchInstitute of Genetic and Developmental BiologyChinese Academy of SciencesShijiazhuang 050021China Key Laboratory of Sugarcane biotechnology and Genetic Improvement(Guangxi) Ministry of Agriculture and Rural AffairsGuangxi Key Laboratory of Sugarcane Genetic ImprovementSugarcane Research InstituteGuangxi Academy of Agricultural SciencesNanning 530007China Queensland Alliance for Agriculture and Food Innovation University of QueenslandSt Lucia 4067Australia

The rapid increase in the proportion of cash crops and livestock production in the Yangtze River Basin has led to commensurate increases in fertilizer and pesticide inputs. Excessive application of chemical fertilizer, organophosphorus pesticides and inappropriate disposal of agricultural waste induced water pollution and potentially threaten Agriculture Green Development(AGD). To ensure food security and the food supply capacity of the Yangtze River Basin, it is important to balance green and development, while ensuring the quality of water bodies. Multiple pollutants affect the transfer, adsorption, photolysis and degradation of each other throughout the soil-plant-water system. This paper considers the impact of multi-pollutants on the nitrogen and phosphorus cycles especially for crops, which are related to achieving food security and AGD. It presents prospective on theory, modeling and multi-pollutant control in the Yangtze River Basin for AGD that are of potential value for other developing regions.

关键词： Agriculture Green Development crop production livestock production multi-pollutant models multiple pollutants Yangtze River Basin

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