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Molecular analysis of the chloroplast Cu/Zn-SOD gene(AhCSD2) in peanut

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The crop Journal 2015年第3期3卷 246-257页

作者： Xiurong Zhang Qian Wan Fengzhen Liu Kun Zhang Aiqing Sun Bing Luo Li Sun Yongshan Wan State Key Laboratory of Crop Biology Shandong Key Laboratory of Crop Biology Shandong Agricultural University

Superoxide dismutase(SOD, EC 1.***.1.1) plays a key role in response to drought stress, and differences in SOD activity changes among cultivars are important under drought conditions. We obtained the full-length DNA of the chloroplast Cu/Zn-SOD gene(Ah CSD2)from 11 allotetraploid cultivars and 5 diploid wild species in peanut. BLAST search against the peanut genome showed that the Ah CSD2 genes g CSD2-1 and g CSD2-2 are located at the tops of chromosome A03(A genome) and B03(B genome), respectively, and both contain 8exons and 7 introns. Nucleotide sequence analyses indicated that g CSD2-2 sequences were identical among all the tested cultivars, while g CSD2-1 sequences showed allelic variations.The amino acid sequences deduced from g CSD2-1 and g CSD2-2 both contain a chloroplast transit peptide and are distinguished by 6 amino acid(aa) residue differences. The other 2aa residue variations in the mature peptide regions give rise to three-dimensional structure changes of the protein deduced from the genes g CSD2-1 and g CSD2-2. Sequences analyses of cultivars and wild species showed that g CSD2-2 of Arachis hypogaea and g Aip CSD2(Arachis ipaensis) are identical, and despite the abundant polymorphic loci between g CSD2-1 of A.hypogaea and sequences from A genome wild species, the deduced amino acid sequence of Ah CSD2-1(A. hypogaea) is identical to that of Adu CSD2(Arachis duranensis), whereas Aco CSD2(Arachis correntina) and Aca CSD2(Arachis cardenasii) both have 2 aa differences in the transit peptide region compared with Ah CSD2-1(A. hypogaea). Based on the Peanut Genome Project, promoter prediction revealed many stress-related cis-acting elements within the potential promoter regions(pp-A and pp-B). pp-A contains more binding sites for drought-associated transcriptional factors than pp-B. We hypothesize that the marked changes in SOD activity in different cultivars under drought stress are tightly regulated by transcription factors through transcription and expression of Ah CSD2 genes.

关键词： Peanut SOD activity AhCSD2 gene Bioinformatics analysis Drought resistance

Characterization and mapping of QTLs on chromosome 2D for grain size and yield traits using a mutant line induced by EMS in wheat

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The crop Journal 2015年第2期3卷 135-144页

作者： Guizhi Zhang Yingying Wang Ying Guo Yan Zhao Fanmei Kong Sishen Li State Key Laboratory of Crop Biology/Shandong key laboratory of crop biology Shandong Agricultural University

Production of mutants with altered phenotypes is a powerful approach for determining the biological functions of genes in an organism. In this study, a high-grain-weight mutant line M8008 was identified from a library of mutants of the common wheat cultivar YN15 treated with ethylmethane sulfonate(EMS). F2 and F2:3generations produced from crosses of M8008 × YN15(MY) and M8008 × SJZ54(MS) were used for genetic analysis. There were significant differences between M8008 and YN15 in plant height(PH), spike length(SL),fertile spikelet number per spike(FSS), grain width(GW), grain length(GL), GL/GW ratio(GLW), and thousand-grain weight(TGW). Most simple correlation coefficients were significant for the investigated traits, suggesting that the correlative mutations occurred in M8008. Approximately 21% of simple sequence repeat(SSR) markers showed polymorphisms between M8008 and YN15, indicating that EMS can induce a large number of mutated loci. Twelve quantitative trait loci(QTLs) forming QTL clusters(one in MY and two in MS) were detected. The QTL clusters coinciding with(MY population) or near(MS population) the marker wmc41 were associated mainly with grain-size traits, among which the M8008 locus led to decreases in GW, factor form density(FFD), and TGW and to increases in GLW. The cluster in the wmc25–barc168 interval in the MS population was associated with yield traits, for which the M8008 locus led to decreased PH, spike number per plant(SN), and SL.

关键词： Common wheat Mutant Simple sequence repeat(SSR) Quantitative trait locus(QTL) Grain size trait Yield trait

Engineered biocontainable RNA virus vectors for non-transgenic genome editing across crop species and genotypes

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Molecular Plant 2023年第3期16卷 616-631页

作者： Qian Liu Chenglu Zhao Kai Sun Yinlu Deng Zhenghe Li state key laboratory of Rice biology Institute of BiotechnologyZhejang UniversityHangzhouChina Ministry of Agricutture key laboratory of Molecular biology of crop Pathogens and Insect Pests Zhejiang UniversityHangzhouChina key laboratory of biology of crop Pathogens and Insects of Zhejang Province Zhejang UniversityHangzhouChina

CRISPR/Cas genome-editing tools provide unprecedented opportunities for basic plant biology research and crop breeding.However,the lack of robust delivery methods has limited the widespread adoption of these revolutionary technologies in plant science.Here,we report an efficient,non-transgenic CRISPR/Cas delivery platform based on the engineered tomato spotted wilt virus(TSWV),an RNA virus with a host range of over 1000 plant species.We eliminated viral elements essential for insect transmission to liberate genome space for accommodating large genetic cargoes without sacrificing the ability to infect plant hosts.The resulting non-insect-transmissible viral vectors enabled effective and stable in planta delivery of Cas12a and Cas9 nucleases as well as adenine and cytosine base editors.In systemically infected plant tissues,the deconstructed TSWV-derived vectors induced efficient somatic gene mutations and base conversions in multiple crop species with little genotype dependency.Plants with heritable,bi-allelic mutations could be readily regenerated by culturing the virus-infected tissues in vitro without antibiotic selection.Moreover,we showed that antiviral treatment with ribavirin during tissue culture cleared the viral vectors in 100%of regenerated plants and further augmented the recovery of heritable mutations.Because many plants are recalcitrant to stable transformation,the viral delivery system developed in this work provides a promising tool to overcome gene delivery bottlenecks for genome editing in various crop species and elite varieties.

关键词： genome editing CRISPR-Cas base editor RNA virus vector tomato spotted wilt virus bunyavirus delivery non-transgenic

The maize late embryogenesis abundant protein Zm DHN13 positively regulates copper tolerance in transgenic yeast and tobacco

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The crop Journal 2019年第3期7卷 403-410页

作者： Yang Liu Daxing Li Qiping Song Tianpeng Zhang Dequan Li Xinghong Yang State Key Laboratory of Crop Biology/Shandong key laboratory of crop biology/College of Life Sciences Shandong Agricultural University

Late embryogenesis abundant (LEA) proteins accumulate in the late stage of plant seed development, and are upregulated in most plants during drought, cold, heat, or salinity stress. LEA proteins can be classified by amino-acid sequence into seven groups. Dehydrins belong to LEA protein group Ⅱ. In previous studies, the maize KS type dehydrin ZmDHN13 increased the tolerance of transgenic tobacco to oxidative stress. In the present study, ZmDHN13 was identified under copper stress conditions, and the protein was then characterized using transgenic yeast and tobacco plants to investigate its functions. ZmDHN13 bound Cu2+. Its overexpression in transgenic tobacco conferred tolerance to copper stress by binding metals and reducing the accumulation of reactive oxygen species (ROS). Three conserved domains displayed a cooperative effect under copper stress conditions.

关键词： Maize LEA proteins Dehydrins ZmDHN13 Copper stress

Disruption of three polyamine uptake transporter genes in rice by CRISPR/Cas9 gene editing confers tolerance to herbicide paraquat

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aBIOTECH 2022年第2期3卷 140-145页

作者： Yu-Shu Lyu Li-Miao Cao Wen-Qian Huang Jian-Xiang Liu Hai-Ping Lu state key laboratory of Plant Physiology and Biochemistry Cllge of Life SciencesZhejang UniversityHangzhou 310058China state key laboratory of Rice biology Zhejang Provincial Key Laboratory of Crop Genetie ResourcesInstitute of Crop ScienceZhejiang UniversityHangzhou 310058China

Weeds are a major biotic constraint that can cause dramatic crop production losses.Herbicide technology has been widely used by farmers as the most cost-effective weed control measure,and development of new strategy to improve herbicide tolerance in plants is urgently needed.The CRISPR/Cas9-based genome editing tool has been used in diverse applications related to agricultural technology for crop improvement.Here we identified three polyamine uptake transporter(PUT)genes in rice that are homologous to the Arabidopsis AtRMV1.We successfully demonstrate that CRISPR/Cas9-targeted mutagenesis of OsPUT1/2/3 greatly improves paraquat resistance in rice without obvious yield penalty.Therefore,manipulation of these loci could be valuable for producing transgene-free rice with improved herbicide resistance in future.

关键词： Paraquat OsPUTs CRISPR/Cas9 Rice

Constructing the barley model for genetic transformation in Triticeae

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Journal of Integrative Agriculture 2015年第3期14卷 453-468页

作者： Lü Bo WU Jia-jie FU Dao-lin State Key Laboratory of Crop Biology/Shandong key laboratory of crop biology Shandong Agricultural University

Barley（Hordeum vulgare L.） is one of the oldest domesticated crops, showing dramatic adaptation to various climate and environmental conditions. As a major cereal crop, barley ranks the 4th after wheat, maize and rice in terms of planting area and production all over the world. Due to its diploid nature, the cultivated barley is considered as an ideal model to study the polyploid wheat and other Triticeae species. Here, we reviewed the development, optimization, and application of transgenic approaches in barley. The most efficient and robust genetic transformation has been built on the Agrobacterium-mediated transfer in conjunction with the immature embryo-based regeneration. We then discussed future considerations of using more practical technologies in barley transformation, such as the T-DNA/transposon tagging and the genome editing. As a cereal crop amenable to genetic transformation, barley will serve as the most valuable carrier for global functional genomics in Triticeae and is becoming the most practical model for generating value-added products.

关键词： Agrobacterium-mediated transfer,genetically modified organism,Hordeum vulgare,immature embryos

Molecular mapping of the hybrid necrosis gene NetJingY176 in Aegilops tauschii using microsatellite markers

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The crop Journal 2015年第4期3卷 298-304页

作者： Fengbo Xue Jun Guo Changying Guan Hongwei Wang Anfei Li Lingrang Kong State Key Laboratory of Crop Biology Shandong Key Laboratory of Crop Biology College of Agronomy Shandong Agricultural University

The rich genetic variation preserved in collections of Aegilops tauschii can be readily exploited to improve common wheat using synthetic hexaploid wheat lines. However,hybrid necrosis, which is characterized by progressive death of leaves or plants, has been observed in certain interspecific crosses between tetraploid wheat and Ae. tauschii. The aim of this study was to construct a fine genetic map of a gene(temporarily named Net Jing Y176)conferring hybrid necrosis in Ae. tauschii accession Jing Y176. A triploid F1 population derived from distant hybridization between Ae. tauschii and tetraploid wheat was used to map the gene with microsatellite markers. The newly developed markers Xsdau K539 and Xsdau K561 co-segregated with Net Jing Y176 on chromosome arm 2DS. The tightly linked markers developed in this study were used to genotype 91 Ae. tauschii accessions. The marker genotype analysis suggested that 49.45% of the Ae. tauschii accessions carry Net Jing Y176. Interestingly, hybrid necrosis genotypes tended to appear more commonly in Ae. tauschii ssp. tauschii than in Ae. tauschii ssp. strangulata.

关键词： Aegilops tauschii Coss Hybrid necrosis Net Jing Y176 Molecular marker

The Puccinia striiformis effector Hasp98 facilitates pathogenicity by blocking the kinase activity of wheat TaMAPK4

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Journal of Integrative Plant biology 2023年第1期65卷 249-264页

作者： Jinping Wei Xiaodong Wang Zeyu Hu Xiaojie Wang Jialiu Wang Jianfeng Wang Xueling Huang Zhensheng Kang Chunlei Tang state key laboratory of crop Stress biology for Arid Areas and College of Plant Protection Northwest A&F University Yangling 712100 China Pioneering Innovation Center for Wheat Stress Tolerance Improvement State Key Laboratory of Crop Stress Biology for Arid Areas Northwest A&F UniversityYangling 712100 China state key laboratory of crop Stress biology for Arid Areas Northwest A&F University Yangling 712100 China

The obligate biotrophic fungus Puccinia striiformis f.***.tritici(Pst)employs virulence effectors to disturb host immunity and causes devastating stripe rust disease.However,our understanding of how Pst effectors regulate host defense responses remains limited.In this study,we determined that the Pst effector Hasp98,which is highly expressed in Pst haustoria,inhibits plant immune responses triggered by flg22or nonpathogenic bacteria.Overexpression of Hasp98 in wheat(Triticum aestivum)suppressed avirulent Pst-triggered immunity,leading to decreased H2O2accumulation and promoting P.striiformis infection,whereas stable silencing of Hasp98 impaired P.striiformis pathogenicity.Hasp98 interacts with the wheat mitogenactivated protein kinase TaMAPK4,a positive regulator of plant resistance to stripe rust.The conserved TEY motif of TaMAPK4 is important for its kinase activity,which is required for the resistance function.We demonstrate that Hasp98inhibits the kinase activity of TaMAPK4 and that the stable silencing of TaMAPK4 compromises wheat resistance against P.striiformis.These results suggest that Hasp98 acts as a virulence effector to interfere with the MAPK signaling pathway in wheat,thereby promoting P.striiformis infection.

关键词： effector MAPK plant immunity Puccinia striiformis f.sp.tritici

Wheat adaptation to environmental stresses under climate change:Molecular basis and genetic improvement

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Molecular Plant 2023年第10期16卷 1564-1589页

作者： Hude Mao Cong Jiang Chunlei Tang Xiaojun Nie Linying Du Yuling Liu Peng Cheng Yunfeng Wu Huiquan Liu Zhensheng Kang Xiaojie Wang state key laboratory of crop Stress biology for Arid Areas and College of Agronomy Northwest A&F UniversityYanglingShaanxi 712100China state key laboratory of crop Stress biology for Arid Areas and College of Plant Protection Northwest A&F UniversityYanglingShaanxi 712100China state key laboratory of crop Stress biology for Arid Areas and College of Life Science Northwest A&F UniversityYanglingShaanxi 712100China

Wheat(Triticum aestivum)is a staple food for about 40%of the world’s population.As the global population has grown and living standards improved,high yield and improved nutritional quality have become the main targets for wheat breeding.However,wheat production has been compromised by global warming through the more frequent occurrence of extreme temperature events,which have increased water scarcity,aggravated soil salinization,caused plants to be more vulnerable to diseases,and directly reduced plant fertility and suppressed yield.One promising option to address these challenges is the genetic improvement of wheat for enhanced resistance to environmental stress.Several decades of progress in genomics and genetic engineering has tremendously advanced our understanding of the molecular and genetic mechanisms underlying abiotic and biotic stress responses in wheat.These advances have heralded what might be considered a“golden age”of functional genomics for the genetic improvement of wheat.Here,we summarize the current knowledge on the molecular and genetic basis of wheat resistance to abiotic and biotic stresses,including the QTLs/genes involved,their functional and regulatory mechanisms,and strategies for genetic modification of wheat for improved stress resistance.In addition,we also provide perspectives on some key challenges that need to be addressed.

关键词： wheat genomics genetic improvement abiotic stress biotic stress climate change