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Molecular Cloning and Functional Characterization of a Salt Tolerance-Associated Gene IbNFU1 from Sweetpotato

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Journal of Integrative Agriculture 2013年第1期12卷 27-35页

作者： WANG Lian-jun HE Shao-zhen ZHAI Hong LIU De-gao WANG Yan-nan LIU Qing-chang key laboratory of crop genomics and genetic improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and UtilizationMinistry of Education/China Agricultural University

Iron-sulfur cluster biosynthesis involving the nitrogen fixation（Nif） proteins has been proposed as a general mechanism acting in various ***-like protein may play an important role in protecting plants against abiotic and biotic *** on the EST sequence selected from salt-stressed suppression subtractive hybridization（SSH） cDNA library constructed with a salt-tolerant mutant LM79,a NFU gene,termed IbNFU1,was cloned from sweetpotato（Ipomoea batatas（L.） Lam.） via rapid amplification of cDNA ends（RACE）.The cDNA sequence of 1 117 bp contained an 846 bp open reading frame encoding a 281 amino acids polypeptide with a molecular weight of 30.5 kDa and an isoelectric point（pI） of ***1 gene contained a conserved Cys-X-X-Cys motif in C-terminal of the iron-sulfur cluster *** deduced amino acid sequence had 66.08 to 71.99% sequence identity to NFU genes reported in Arabidopsis thaliana,Eucalyptus grandis and Vitis ***-time quantitative PCR analysis revealed that the expression level of IbNFU1 gene was significantly higher in the roots of the mutant LM79 compared to the wild-type *** tobacco（*** 38） plants expressing IbNFU1 gene exhibited significantly higher salt tolerance compared to the untransformed control *** is proposed that IbNFU1 gene has an important function for salt tolerance of plants.

关键词： cloning IbNFU1 gene Ipomoea batatas(L.) Lam. salt tolerance

Molecular mapping of QTLs for root response to phosphorus deficiency at seedling stage in wheat (Triticum aestivum L.)

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Progress in Natural Science:Materials International 2007年第10期17卷 1177-1184页

作者： Li Zhenxing, Ni Zhongfu, Peng Huiru, Liu Zhiyong, Nie Xiuling, Xu Shengbao, Liu Gang and Sun Qixin(key laboratory of crop Heterosis and Utilization and State key laboratory for Agrobiotechnology, key laboratory of crop genomics and genetic improvement, Beijing key laboratory of crop genetic improvement, Department of Plant genetics & Breeding, China Agricultural University, Beijing 100094, China) key laboratory of crop Heterosis and Utilization and State key laboratory for Agrobiotechnology Key Laboratory of Crop Genomics and Genetic Improvement Beijing Key Laboratory of Crop Genetic Improvement Department of Plant Genetics & Breeding China Agricultural University Beijing 100094 China

Phosphorus (P) deficiency in the soil is one of the major abiotic stresses that limit plant growth and crop productivity throughout the world. Development of cultivars with improved P-deficiency tolerance is an efficient strategy for sustainable agriculture. Plant roots play an important role in crop growth and development, especially in nutrient uptake and improvement of P-efficiency. Mapping quantitative trait loci (QTLs) for root traits and their response to low P stress at seedling stage will facilitate the development of P-efficient wheat cultivars. In this study, 30 QTLs (LOD>2.0) were mapped for the three root traits, such as root length, root number and root dry matter under different P supply conditions and their response to P-stress. These QTLs were distributed on 14 chromosomes, with each of the 5 QTLs explaining more than 10% phenotype variance. Analyses showed that root traits and their response to P-deficiency were controlled by different QTLs. In addition, alleles with positive effects were separated on both parents, and wheat cultivars with improved P-efficiency could be developed by accumulating these positive effect alleles together.

关键词： QTLs roots wheat seedling RIL P-deficiency.

Wheat 14-3-3 Protein Conferring Growth Retardation in Arabidopsis

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Journal of Integrative Agriculture 2013年第2期12卷 209-217页

作者： LI Jing SONG Su-sheng ZHAO Yu-sheng GUO Wei-wei GUO Guang-hui PENG Hui-ru NI Zhong-fu SUN Qi-xin YAO Ying-yin State key laboratory for Agrobiotechnology and key laboratory of crop Heterosis and Utilization Minstry of Education/Key Laboratory of Crop Genomics and Genetic Improvement Minstry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University School of Life Sciences Tsinghua University

14-3-3 proteins belong to a family of phosphoserine/threonine-binding modules and participate in a wide array of signal transduction and regulatory events. Our previous study demonstrated that Ta14-3-3 was significantly down-regulated in leaf and root tissues of hybrid wheat at the tillering stage. In this paper, three homoeologous Ta14-3-3 genes were cloned from common wheat （Triticum aestivum L., 2n=6x=42, AABBDD） and mapped on chromosomes 2A, 2B, and 2D, respectively. Transgenic Arabidopsis plants ectopically overexpressing Ta14-3-3 displayed shorter primary roots, delayed flowering and retarded growth rates, indicating that Ta14-3-3 acted as a growth inhibitor in Arabidopsis. In wheat, Ta14-3-3 was down-regulated in roots and leaves of hybrids as compared to their parental lines. We proposed that Ta14-3-3 proteins might regulate growth vigor in hybrid wheat.

关键词： wheat Ta14-3-3 heterosis gene expression

Cloning and characterization of an up-regulated GA 20-oxidase gene in hybrid maize

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Progress in Natural Science:Materials International 2009年第2期19卷 161-166页

作者： Jinkun Du,Yingyin Yao,Zhongfu Ni,Qixin Sun Department of Plant genetics & Breeding and State key laboratory for Agrobiotechnology,key laboratory of crop genomics and genetic improvement,Ministry of Agriculture/Beijing key laboratory of crop genetic improvement,China Agricultural University,Beijing 100094,China [a]Department of Plant genetics & Breeding and State key laboratory for Agrobiotechnology Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University Beijing 100094 China

Previous studies revealed that GA content and metabolism are positively correlated with a faster shoot growth rate of hybrid,and recently,genes participating in both GA biosynthesis and GA response pathways were also found to be differentially expressed between wheat hybrid and its parental inbreds. In this study,an up-regulated GA 20-oxidase gene in a maize hybrid,designated ZmGA20,was cloned. ZmGA20 contains an open reading frame (ORF) encoding 391 amino acid residues. BLASTX searches in GenBank revealed that the ZmGA20 is homologous to the sequences of GA20ox proteins from different species,and analysis also indicated that ZmGA20 had typical features of GA 20-oxidase proteins,including a "LPWKET" sequence. Semi-quantitative RT-PCR analysis showed that ZmGA20 was expressed in different tissues and/or organs. The expression level of ZmGA20 in the hybrid was higher than that in two parents (in roots,leaves,stems and embryo,and ears). The abundance of ZmGA20 transcript was equal to that of the highly expressed parents,which provided molecular evidence for the observed GA content heterosis in maize hybrids.

关键词： GA 20-oxidase Gene expression Maize Heterosis

Enhanced Stem Nematode Resistance of Transgenic Sweetpotato Plants Expressing Oryzacystatin-I Gene

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Agricultural Sciences in China 2011年第4期10卷 519-525页

作者： GAO Shang YU Bo ZHAI Hong HE Shao-zhen LIU Qing-chang key laboratory of crop genomics and genetic improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement/Laboratory of Crop Heterosis and Utilization Ministry of Education~China Agricultural University Beijing 100193 P.R.China

Enhanced stem nematode resistance of transgenic sweetpotato （cv. Lizixiang） was achieved using Oryzacystatin-I （OCI） gene with Agrobacterium tumefaciens-mediated transformation. A. tumefaciens strain EHA105 harbors a binary vector pCAMBIA1301 with OCI gene, gusA gene and hptII gene. Selection culture was conducted using 25 mg L-1 hygromycin. A total of 1 715 plants were produced from the inoculated 1 450 cell aggregates of Lizixiang via somatic embryogenesis. GUS assay and PCR analysis of the putative transgenic plants randomly sampled showed that 90.54% of them were transgenic plants. Transgenic plants exhibited significantly enhanced resistance to stem nematodes compared to the untransformed control plants by the field evaluation with stem nematodes. Stable integration of the OCI gene into the genome of resistant transgenic plants was confirmed by Southern blot analysis, and the copy number of integrated OCI gene ranged from 1 to 4. Transgene overexpression in stem nematode-resistant plants was demonstrated by quantitative real-time PCR analysis. This study provides a way for improving stem nematode resistance in sweetpotato.

关键词： Agrobacterium tumefaciens Ipomoea batatas (L.) Lam. Oryzacystatin-I gene stem nematode resistance transgenic plant

Genomic and genic sequence variation in synthetic hexaploid wheat (AABBDD) as compared to their parental species

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Progress in Natural Science:Materials International 2008年第5期18卷 533-538页

作者： Lihong Nie, Zongfu Han, Lahu Lu, Yingyin Yao, Qixin Sun, Zhongfu Ni key laboratory of crop Heterosis and Utilization (MOE) and State key laboratory for Agrobiotechnology, key laboratory of crop genomics and genetic improvement (MOA), Beijing key laboratory of crop genetic improvement, Department of Plant genetics & Breeding, China Agricultural University, Beijing 100094, China key laboratory of crop Heterosis and Utilization (MOE) and State key laboratory for Agrobiotechnology Key Laboratory of Crop Genomics and Genetic Improvement (MOA) Beijing Key Laboratory of Crop Genetic Improvement Department of Plant Genetics & Breeding China Agricultural University Beijing 100094 China

In order to understand the genomic changes during the evolution of hexaploid wheat, two sets of synthetic hexaploid wheat from hybridization between maternal tetraploid wheat (AABB) and paternal diploid goat grass (DD) were used for DNA-AFLP and single strand conformation polymorphism (SSCP) analysis to determine the genomic and genic variation in the synthetic hexaploid wheat. Results indicated that more DNA sequences from paternal diploid species were eliminated in the synthetic hexaploid wheat than from maternal tetraploid wheat, suggesting that genome from parental species of lower ploidity tends to be eliminated preferentially. However, sequence variation detected by SSCP procedure was much lower than those detected by DNA-AFLP, which indicated that much less variation in the genic regions occurred in the synthetic hexaploid wheat, and sequence variations detected by DNA-AFLP could be derived mostly from non-coding regions and repetitive sequences. Our results also indicated that sequence variation in 4 genes can be detected in hybrid F1, which suggested that this type of sequence variation could be resulted from distant hybridization. It was interesting to note that 3 out of the 4 genes were mapped and clustered on the long arm of chromosome 2D, which indicated that variation in genic sequences in synthetic hexaploid wheat might not be a randomized process.

关键词： Synthetic hexaploid wheat Polyploidy Genome Sequence variation

Ectopic Overexpression of Wheat Adenosine Diphosphate-ribosylation Factor,TaARF,Increases Growth Rate in Arabidopsis

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Journal of Integrative Plant Biology 2009年第1期51卷 35-44页

作者： Yingyin Yao Zhongfu Ni Jinkun Du Zongfu Han Yan Chen Qingbo Zhang Qixin Sun Department of Plant genetics & Breeding and State key laboratory for Agrobiotechnology Key Laboratory of Crop Heterosis and UtilizationMinistry of Education Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agdculture Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University Beijing 100094 China

Differential gene expression between hybrids and their parents is considered to be associated with heterosis. However, the physiological functions and possible contribution to heterosis of these differentially expressed genes are unknown. We have isolated one hybrid upregulated gene encoding putative wheat ADP-ribosylation factor, designated TaARF. In this study, real-time quantitative reverse transcription-polymerase chain reaction analysis indicated that the TaARF transcript was preferentially expressed in root, node and crown, and the accumulation of TaARF mRNA in hybrid was more than 1.5-fold higher than that in two parents. In order to understand possible roles of the putative wheat ARF gene, TaARF was overexpressed in Arabidopsis, and the transgenic plants were characterized. We show that ectopic overexpression of TaARF in Arabidopsis leads to increased leaf area, increased growth rate and earlier transition to flowering, suggesting that TaARF plays significant roles in growth and development. This study provides evidence demonstrating that TaARF plays important roles in growth and development and we speculate that the upregulated expression of this gene might contribute to the heterosis observed in wheat root and leaf growth.

关键词： adenosine diphosphate-ribosylation factor Arabidopsis ectopic heterosis overexpression wheat.

Cloning and expression profiles of 15 genes encoding WRKY transcription factor in wheat (Triticum aestivem L.)

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Progress in Natural Science:Materials International 2008年第6期18卷 697-705页

作者： Hualing Wu,Zhongfu Ni,Yingyin Yao,Ganggang Guo,Qixin Sun* Department of Plant genetics&Breeding and State key laboratory for Agrobiotechnology,key laboratory of crop Heterosis and Utilization,Ministry of Education,key laboratory of crop genomics and genetic improvement,Ministry of Agriculture/Beijing key laboratory of crop genetic improvement,China Agricultural University,Beijing 100094,China Department of Plant genetics & Breeding and State key laboratory for Agrobiotechnology Key Laboratory of Crop Heterosis and Utilization Ministry of Education Key Laboratory of Crop Genomics and Genetic Improvement Ministry of Agriculture/Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University Beijing 100094 China

WRKY proteins are involved in various physiological processes,including biotic and abiotic stress responses,hormone responses and ***,no systematic identification,expression and function analysis of WRKY genes in wheat were *** this study,we isolated 15 wheat cDNAs with complete open reading frame(ORF)encoding putative WRKY proteins using in silico *** analysis indicated that the 15 wheat WRKY genes belonged to three major WRKY *** analysis revealed that most genes expressed drastically in leaf,except TaWRKY10 which expressed in crown *** genes were strongly up-reg-ulated with the senescence of *** genes were responsive to low temperature,high temperature,NaCl or PEG ***-over,dierential expression patterns were also observed between wheat hybrid and its parents,and some genes were more responsive to PEG treatment in the *** results demonstrated that wheat WRKY genes are involved in leaf senescing and abiotic *** the changed expression of these WRKY genes in hybrid might contribute to the heterosis by improving the stress tolerance in hybrids.

关键词： Wheat WRKY transcription factor Gene cloning Gene expression Heterosis

Comparative transcriptome profiling of two maize near-isogenic lines differing in the allelic state for bacterial brown spot disease resistance

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Journal of Integrative Agriculture 2015年第4期14卷 610-621页

作者： WU Xiao-jun Xu Li ZHAO Pan-feng LI Na WU Lei HE Yan WANG Shou-cai National Maize improvement Center of China/key laboratory of crop genomics and genetic of Maize China Agricultural University

The bacterial brown spot disease（BBS）, caused primarily by Pseudomonas syringae pv. syringae van Hall（Pss）, reduces plant vigor, yield and quality in maize. To reveal the nature of the defense mechanisms and identify genes involved in the effective host resistance, the dynamic changes of defense transcriptome triggered by the infection of Pss were investigated and compared between two maize near-isogenic lines（NILs）. We found that Pss infection resulted in a sophisticated transcriptional reprogramming of several biological processes and the resistant NIL employed much faster defense responses than the susceptible NIL. Numerous genes encoding essential components of plant basal resistance would be able to be activated in the susceptible NIL, such as PEN1, PEN2, PEN3, and EDR1, however, in a basic manner, such resistance might not be sufficient for suppressing Pss pathogenesis. In addition, the expressions of a large number of PTI-, ETI-, PR-, and WRKY-related genes were pronouncedly activated in the resistant NIL, suggesting that maize employ a multitude of defense pathways to defend Pss infection. Six R-gene homologs were identified to have significantly higher expression levels in the resistant NIL at early time point, indicating that a robust surveillance system（gene-to-gene model） might operate in maize during Pss attacks, and these homolog genes are likely to be potential candidate resistance genes involved in BBS disease resistance. Furthermore, a holistic group of novel pathogen-responsive genes were defined, providing the repertoire of candidate genes for further functional characterization and identification of their regulation patterns during pathogen infection.

关键词： maize(Zea mays L.) bacterial brown spot disease RNA-Seq analysis disease resistance