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CRISPR/CasRx-mediated resistance to Soybean mosaic virus in soybean

The Crop Journal 2024年第4期12卷 1093-1101页

作者： Le gao Lijun Xie Yanmin Xiao Xinge Cheng Ruosi Pu Ziheng Zhang Yu Liu shaopei gao Zilong Zhang Haoran Qu Haijian Zhi Kai Li MARA Key Laboratory of Biology and Genetics Improvement of Soybean Zhongshan Biological Breeding Laboratory(ZSBBL)National Innovation Platform for Soybean Breeding and Industry-Education IntegrationState Key Laboratory of Crop Genetics&Germplasm Enhancement and UtilizationNanjing Agricultural UniversityNanjing 210095JiangsuChina Department of Horticulture Beijing Vocational College of AgricultureBeijing 102442China College of Agronomy&Biotechnology China Agricultural UniversityBeijing 100193China College of Bioscience and Biotechnology Shenyang Agricultural UniversityShenyang 110866LiaoningChina

Soybean mosaic virus(SMV),an RNA virus,is the most common and destructive pathogenic virus in soybean *** newly developed CRISPR/Cas immune system has provided a novel strategy for improving plant resistance to viruses;hence,this study aimed to engineer SMV resistance in soybean using this ***,multiple sgRNAs were designed to target positive-and/or negative-sense strands of the SMV HC-Pro ***,the corresponding CRISPR/CasRx vectors were constructed and transformed into *** inoculation with SMV,39.02%,35.77%,and 18.70%of T_(1)plants were confirmed to be highly resistant(HR),resistant(R),and mildly resistant(MR)to SMV,respectively,whereas only 6.50%were identified as susceptible(S).Additionally,qRT-PCR and DAS-ELISA showed that,both at 15 and 30 d post-inoculation(dpi),SMV accumulation significantly decreased or was even undetectable in HR and R plants,followed by MR and S ***,the expression level of the CasRx gene varied in almost all T_(1)plants with different resistance level,both at 15 and 30 ***,when SMV resistance was evaluated in the T_(2)generation,the results were similar to those recorded for the T_(1)*** findings provide new insights into the application of the CRISPR/CasRx system for soybean improvement and offer a promising alternative strategy for breeding for resistance to biotic stress that will contribute to the development of SMV-immune soybean germplasm to accelerate progress towards greater soybean crop productivity.

关键词： Soybean RNA virus Soybean mosaic virus Resistance CRISPR/CasRx

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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 *** abiotic stress tolerance holds significant importance in fully utilizing marginal *** processes regulate abiotic stress responses,yet the molecular regulatory mechanisms in sweet potato remain *** this study,a NAC(NAM,ATAF1/2,and CUC2)transcription factor,IbNAC087,was identified,which is commonly upregulated in salt-and drought-tolerant *** 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 ***-rich IbNAC087-OE(overexpression)plants exhibited more stomatal closure than wild-type(WT)and IbNAC087-Ri plants under NaCl,polyethylene glycol,and methyl jasmonate ***087 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 *** ubiquitinated IbNAC087 to promote 26S proteasome degradation,which weakened its activation on IbLOX and *** 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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Fine-Tuning of Euil： Breaking the Bottleneck in Hybrid Rice Seed Production

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Molecular Plant 2018年第5期11卷 643-644页

作者： shaopei gao Chengcai Chu State Key Laboratory of Plant Genomics and National Center for Plant Gene Research (Beijing) Institute of Genetics and Developmental Biology Chinese Academy of Sciences Beijing 100101 China

Gibberellins （GAs） are essential phytohormones regulating many aspects of plant growth and development. Manipulation of GA status either by genetic alteration or by exogenous application of GA or GA biosynthesis inhibitors are common strategies used to optimize plant growth and crop yields. Rice （Oryza sativa） is a major source of calories and mineral nutrients, feeding more than half of the human population. With the challenges of feeding an increasing population and decreasing availability of arable land, improving productivity and yields of rice crop has always been an ultimate goal for breeders.

关键词： CYTOCHROME-P450 MONOOXYGENASE PANICLE EXSERTION ELEMENT TARGET GENE

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Development of a high-density SSR genetic linkage map in sweet potato

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The Crop Journal 2021年第6期9卷 1367-1374页

作者： Yusha Meng Chenxing Zheng Hui Li Aixian Li Hong Zhai Qingmei Wang Shaozhen He Ning Zhao Huan Zhang shaopei gao Qingchang Liu Key Laboratory of Sweetpotato 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 and BiotechnologyChina Agricultural UniversityBeijing 100193China Crop Research Institute Shandong Academy of Agricultural SciencesJinan 250100ShandongChina

Simple sequence repeat(SSR) markers have been proved to be a very powerful tool for quantitative trait locus(QTL) mapping, marker-assisted selection and comparative genomics research in many crop species. However, a high-density SSR genetic linkage map is still lacking because there are only a few SSR markers available in sweet potato. In this study, a total of 2545 simple sequence repeat(SSR) primer pairs, including 1215 genomic SSR(gSSR) primer pairs and 1330 BES-SSR(bSSR) primer pairs designed from the genome sequence and BAC-end sequence of sweet potato, respectively, were screened with sweet potato cultivars Luoxushu 8 and Zhengshu 20 and their randomly sampled two F1 individuals and 571 of them generated polymorphic bands. The selected 571 polymorphic SSR primer pairs and 35 EST-based SSR(eSSR) primer pairs developed at our laboratory were used to genotype 240 F1 individuals derived from a cross between Luoxushu 8 and Zhengshu 20. A double pseudo-test-cross strategy was applied for linkage analysis. The Luoxushu 8 map included 90 linkage groups with 5057 SSR markers and covered 13,299.9 cM with a marker density of 2.6 cM, and the Zhengshu 20 map contained 90 linkage groups with 3009 SSR markers and covered 11,122.9 cM with a marker density of 3.7 cM. Fifteen homologous groups were identified in both parent maps. These are the first SSR linkage maps consisting of the complete 90 linkage groups and 15 homologous groups, which are consistent with the autohexaploid nature of sweetpotato, and are also the linkage maps with the highest SSR marker density reported to *** results provide a basis for QTL mapping, marker-assisted breeding and comparative genomics research of sweet potato.

关键词： Sweet potato SSR Pseudo-test-cross strategy Linkage map Autohexaploid

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Big Grain3, encoding a purine permease, regulates grain size via modulating cytokinin transport in rice

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Journal of Integrative Plant Biology 2019年第5期61卷 581-597页

作者： Yunhua Xiao Dapu Liu Guoxia Zhang shaopei gao Linchuan Liu Fan Xu Ronghui Che Yiqin Wang Hongning Tong Chengcai Chu State Key Laboratory of Plant Genomics and National Center for Plant Gene Research(Beijing) Institute of Genetics and Developmental Biologythe Chinese Academy of SciencesBeijing 100101China National Key Facility for Crop Gene Resources and Genetic Improvement Institute of Crop Sciencesthe Chinese Academy of Agricultural SciencesBeijing 100081China

Grain size is an important agronomic trait affecting grain yield,but the underlying molecular mechanisms remain to be ***,we isolated a dominant mutant,big grainy(bg3-D),which exhibits a remarkable increase of grain size caused by activation of the PURINE PERMEASE gene,***3/OSPUP4 is predominantly expressed in vascular tissues and is specifically suppressed by exogenous cytokinin *** profiling revealed that the distribution of different cytokinin forms,in roots and shoots of the bg3-D mutant,is *** reverse transcription-PCR(qRT-PCR)analysis indicated that expression of rice cytokinin type-A RESPONSE REGULATOR(OsRR)genes is enhanced in the roots of the bgj-D *** results suggest that OSPUP4 might contribute to the long-distance transport of cytokinin,by reinforcing cytokinin loading into vascular bundle ***,plants overexpressing OsPUP7,the closest homolog of OsPUP4,also exhibited a similar phenotype to the bg3-D ***,subcellular localization demonstrated that OSPUP4 was localized on the plasma membrane,whereas OSPUP7 was localized to the endoplasmic *** on these findings,we propose that OSPUP4 and OSPUP7 function in a linear pathway to direct cytokinin cell-to-cell transport,affecting both its long-distance movement and local allocation.

关键词： Grain size is an important underlying molecular local allocation

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Overexpression of micro RNA_(40)8 enhances photosynthesis, growth, and seed yield in diverse plants

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Journal of Integrative Plant Biology 2018年第4期60卷 323-340页

作者： Jiawei Pan Dahui Huang Zhonglong Guo Zheng Kuang He Zhang Xinyu Xie Zengfeng Ma shaopei gao Manuel T.Lerdau Chengcai Chu Lei Li State Key Laboratory of Protein and Plant Gene Research Peking-Tsinghua Center for Life SciencesSchool of Life Sciences and School of Advanced Agricultural SciencesPeking University Rice Research Institute Guangxi Academy of Agricultural Sciences State Key Laboratory of Plant Genomics and National Center for Plant Gene Research(Beijing) Institute of Genetics and Developmental BiologyChinese Academy of Sciences Department of Environmental Sciences University of Virginia

The ability of a plant to produce grain, fruit, or forage depends ultimately on photosynthesis. There have been few attempts, however, to study microRNAs, which are a class of endogenous small RNAs post-transcription- ally programming gene expression, in relation to photosynthetic traits. We focused on miR408, one of the most conserved plant miRNAs, and overexpressed it in parallel in Arabidopsis, tobacco, and rice. The transgenic plants all exhibited increased copper content in the chloroplast, elevated abundance of plastocyanin, and an induction of photosynthetic genes. By means of gas exchange and optical spectroscopy analyses, we showed that higher expression of miR408 leads to enhanced photosynthesis through improving efficiency of irradiation utilization and the capacity for carbon dioxide fixation. Consequently, miR408 hyper-accumulat- ing plants exhibited higher rate of vegetative growth. An enlargement of seed size was also observed in all three species overproducing miR408. Moreover, we conducted a 2-year-two-location field trial and observed miR408 overexpression in rice significantly increased yield, which was primarily attributed to an elevation in grain weight. Taken together, these results demonstrate that miR408 is a positive regulator of photosynthesis and that its genetic engineering is a promising route for enhancing photosynthetic performance and yield in diverse plants.

关键词： and seed yield in diverse plants growth Figure

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Construction of a high-density SSR genetic linkage map and identification of QTL for storage-root yield and dry-matter content in sweetpotato

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The Crop Journal 2023年第3期11卷 963-967页

作者： Chenxing Zheng Zhicheng Jiang Yusha Meng Jun Yu Xinsun Yang Huan Zhang Ning Zhao Shaozhen He shaopei gao Hong Zhai Qingchang Liu Key Laboratory of Sweetpotato Biology and Biotechnology Ministry of Agriculture and Rural Affairs/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis&UtilizationMinistry of EducationCollege of Agronomy&BiotechnologyChina Agricultural UniversityBeijing 100193China Institute of Rural Development Zhejiang Academy of Agricultural SciencesHangzhou 310021ZhejiangChina Institute of Food Crops Hubei Academy of Agricultural SciencesWuhan 430064HubeiChina

Sweetpotato(Ipomoea batatas(L.)Lam.)is a widely grown food crop especially in developing *** storage-root yield and dry-matter content has been the main breeding objective of the crop,and DNA marker-assisted breeding is needed for this *** this study,using a mapping population of 500 F1 individuals from a cross between Xushu 18(female)and Xu 781(male),we constructed a highdensity genetic linkage map of sweetpotato using 601 simple-sequence repeat(SSR)primer *** Xushu 18 map contained 90 linkage groups with 5547 SSR markers and spanned 18,263.5 cM,and the Xu 781 map contained 90 linkage groups with 4599 SSR markers and spanned 18,043.7 cM,representing the highest genome coverage yet reported for *** identified 33 QTL for storage-root yield and 16 QTL for dry-matter content,explaining respectively 6.5%–47.5%and 3.2%–18.9%of *** results provide a foundation for fine-mapping and cloning of QTL and for marker-assisted breeding in sweetpotato.

关键词： Sweetpotato SSR linkage map QTL Storage-root yield Dry-matter content

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A conserved unusual posttranscriptional processing mediated by short,direct repeated (SDR) sequences in plants

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Journal of Genetics and Genomics 2010年第1期37卷 85-99页

作者： Xiangli Niu Di Luo shaopei gao Guangjun Ren Lijuan Chang Yuke Zhou Xiaoli Luo Yuxiang Li Pei Hou Wei Tang Bao-Rong Lu Yongsheng Liu College of Agricultural and Life Sciences Chongqing UniversityChongqing 400030China Ministry of Education Key Laboratory for Bio-resource and Eco-environment College of Life Science and State Key Laboratory of BiotherapySichuan UniversityChengdu 610064China c Institute of Crop Research Sichuan Academy of Agricultural SciencesChengdu 610066China Ministry of Education Key Laboratory for Biodiversity Science and Ecological Engineering Institute of Biodiversity ScienceFudan UniversityShanghai 200433China1

In several stress responsive gene loci of monocot cereal crops,we have previously identified an unusual posttranscriptional processing mediated by paired presence of short direct repeated （SDR） sequences at 5＇ and 3＇ splicing junctions that are distinct from conventional （U2/U12-type） splicing *** using the known SDR-containing sequences as probes,24 plant candidate genes involved in diverse functional pathways from both monocots and dicots that potentially possess SDR-mediated posttranscriptional processing were predicted in the GenBank *** SDRs-mediated posttranscriptional processing events including cis-and trans-actions were experimentally detected in majority of the predicted *** sequence analysis demonstrates several types of SDR-associated splicing peculiarities including partial exon deletion,exon fragment repetition,exon fragment scrambling and trans-splicing that result in either loss of partial exon or unusual exonic sequence rearrangements within or between RNA *** addition,we show that the paired presence of SDR is necessary but not sufficient in SDR-mediated splicing in transient expression and stable transformation *** also show prokaryote is incapable of SDR-mediated premRNA splicing.

关键词： posttranscriptional processing short direct repeat (SDR) premRNA splicing plant

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The transcription factor IbNAC29 positively regulates the carotenoid accumulation in sweet potato

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Horticulture Research 2023年第3期10卷 189-200页

作者： Shihan Xing Ruijie Li Haoqiang Zhao Hong Zhai Shaozhen He Huan Zhang Yuanyuan Zhou Ning Zhao shaopei gao Qingchang Liu 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 and UtilizationMinistry of EducationCollege of Agronomy&BiotechnologyChina Agricultural UniversityBeijing 100193China

Carotenoid is a tetraterpene pigment beneficial for human *** the carotenoid biosynthesis pathway has been extensively studied in plants,relatively little is known about their regulation in sweet ***,we conducted the transcriptome database of differentially expressed genes between the sweet potato(Ipomoea batatas)cultivar‘Weiduoli’and its high-carotenoid mutant‘HVB-3’.In this study,we selected one of these candidate genes,IbNAC29,for subsequent ***29 belongs to the plant-specific NAC(NAM,ATAF1/2,and CUC2)transcription factor *** IbNAC29 mRNA level in the HVB-3 storage roots was∼1.71-fold higher than *** experiments showed that the contents ofα-carotene,lutein,β-carotene,zeaxanthin,and capsanthin are obviously increased in the storage roots of transgenic sweet potato plants overexpressing ***,the levels of carotenoid biosynthesis genes in transgenic plants were also ***,yeast one-hybrid assays indicated that IbNAC29 could not directly bind to the promoters of these carotenoid biosynthesis ***,the level of IbSGR1 was down-regulated,whose homologous genes in tomato can negatively regulate carotene *** three-hybrid analysis revealed that the IbNAC29-IbMYB1R1-IbAITR5 could form a regulatory *** one-hybrid,electrophoretic mobility shift assay,quantitative PCR analysis of chromatin immunoprecipitation and dual-luciferase reporter assay showed that IbAITR5 directly binds to and inhibits the promoter activity of IbSGR1,up-regulating carotenoid biosynthesis gene *** together,IbNAC29 is a potential candidate gene for the genetic improvement of nutritive value in sweet potato.

关键词： roots sweet potato

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植物油体蛋白基因家族研究进展

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遗传 2022年第12期44卷 1128-1140页

作者：赵浩强王小斐高少培中国农业大学教育部作物杂种优势研究与利用重点实验室/农业农村部甘薯生物学与生物技术重点实验室北京100193 中国农业大学食品科学与营养工程学院北京100083

油体也称脂滴或油滴,是植物细胞中一种重要的储藏油脂的细胞器。油体由单层磷脂膜包裹中性脂肪酸组成,膜上镶嵌有多种膜蛋白,包括油体蛋白、油体钙蛋白和油体固醇蛋白,其中油体蛋白占80%~90%。油体蛋白在影响油体大小与稳定性、油体形... 详细信息

油体也称脂滴或油滴,是植物细胞中一种重要的储藏油脂的细胞器。油体由单层磷脂膜包裹中性脂肪酸组成,膜上镶嵌有多种膜蛋白,包括油体蛋白、油体钙蛋白和油体固醇蛋白,其中油体蛋白占80%~90%。油体蛋白在影响油体大小与稳定性、油体形成和降解、脂质代谢、种子成熟及萌发等多种生命活动中都发挥着重要的生物学作用。本文结合近些年国内外关于植物油体蛋白基因家族的研究进展系统总结了油体蛋白序列与结构特征及在植物生长发育中所扮演的重要角色,讨论了油体蛋白作为一种新型植物蛋白在实际生产中的应用场景和油体蛋白研究及应用过程中仍存在的一些问题,以期为人们后期更加深入研究植物油体蛋白相关分子功能及在生产实践中应用提供有益的参考。

关键词：油体油体蛋白油体稳定性脂质代谢油脂合成

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