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Mapping of three QTLs for seed setting and analysis on the candidate gene for qSS-1 in rice(Oryza sativa L.)

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Journal of Integrative Agriculture 2016年第4期15卷 735-743页

作者： Elsheikh Y M Ahmed ZHANG Yan-pei YU Jian-ping Rashid M A Rehman ZHANG Zhan-ying ZHANG Hong-liang LI Jin-jie LI Zi-chao key laboratory of crop heterosis and utilization Ministry of Education Beijing Key Laboratory of Crop Genetic Improvement China Agricultural University

The lower seed setting is one of the major hindrances which face grain yield in rice. One of the main reasons to cause low spikelet fertility （seed setting） is male sterility or pollen abortion. Notably, pollen abortion has been frequently observed in advanced progenies of rice. In the present study, 149 BC2F6 individuals with significant segregation in spikelet fertility were generated from the cross between N040212 （indica） and Nipponbare （japonica） and used for primary gene mapping. Three QTLs, qSS-1, qSS-7 and qSS-9 at chromosomes 1, 7 and 9, respectively, were found to be associated with seed setting. The recombinant analysis and the physical mapping information from publicly available resources exhibited that the qSS-1, qSS-7 and qSS-9 loci were mapped to an interval of 188, 701 and 3741 kb, respectively. The seed setting responsible for QTL qSS-1 was further fine mapped to 93.5 kb by using BC2F7 population of 1 849 individuals. There are 16 possible putative genes in this 93.5 kb region. Pollen vitality tests and artificial pollination indicated that the male gamete has abnormal pollen while the female gamete was normal. These data showed that low seed setting rate relative to qSS-1 may be caused by abnormal pollen grains. These results will be useful for cloning, functional analysis of the target gene governing spikelet fertility （seed setting） and understanding the genetic bases of pollen sterility.

关键词： rice (Oryza sativa L.) QTL mapping seed setting pollen sterility

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

Overexpression of IbMIPS1 gene enhances salt tolerance in transgenic sweetpotato

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Journal of Integrative Agriculture 2016年第2期15卷 271-281页

作者： WANG Fei-bing ZHAI Hong AN Yan-yan SI Zeng-zhi HE Shao-zhen LIU Qing-chang beijing key laboratory of crop genetic improvement/laboratory of crop heterosis and utilization Ministry of EducationChina Agricultural University

Myo-inositol-1-phosphate synthase（MIPS） is a key rate limiting enzyme in the de novo biosynthesis of myo-inositol in *** the present study,the IbMIPS1 gene was introduced into sweetpotato cultivar Xushu 18 and the transgenic plants exhibited significantly enhanced salt tolerance compared with the wild-type（WT）.Overexpression of IbMIPSI up-regulated the salt stress responsive genes,including myo-inositol monophosphatase（MIPP）,pyrroline-5-carboxylate synthase（P5CS）,pyrroline-5-carboxylate reductase（P5CR）,psbA,phosphoribulokinase（PRK）,and superoxide dismutase（SOD） genes,under salt *** and proline content,SOD and photosynthesis activities were significantly increased,whereas malonaldehyde（MDA） and H_2O_2 contents were significantly decreased in the transgenic *** findings suggest that the IbMIPS1 gene may enhance salt tolerance of sweetpotato by regulating the expression of salt stress responsive genes,increasing the content of inositol and proline and enhancing the activity of photosynthesis.

关键词： IbMIPS1 overexpression salt tolerance sweetpotato

Identification of Differentially Expressed Genes in Sweetpotato Storage Roots Between Kokei No.14 and Its Mutant Nongdafu 14 Using PCR-Based cDNA Subtraction

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Journal of Integrative Agriculture 2013年第4期12卷 589-595页

作者： CHEN Wei ZHAI Hong YANG Yuan-jun HE Shao-zhen LIU De-gao LIU Qing-chang beijing key laboratory of crop genetic improvement/laboratory of crop heterosis and utilization Ministry of Education/China Agricultural University

The contents of earotenoids in the storage root of sweetpotato, Ipomoea batatas （L.） Lam. vary dramatically among different cultivars. However, so far little is known about the regulation of carotenoids synthesis in sweetpotato. In our laboratory, we identified a novel sweetpotato mutant, Nongdafu 14, which is a homogenous mutant derived from the wild type Kokei No. 14. The contents of carotenoids in the storage root of Nongdafu 14 were analyzed using high performance liquid chromatography （HPLC）, and it was found that the amount of carotenoids, [3-carotene, lutein and zeaxantion, three major types of earotenoids in sweetpotato storage roots, increased 2-26 folds in Nongdafu 14 compared to Kokei No. 14. Suppression subtractive hybridization （SSH） was used to identify genes that were differentially expressed in Nongdafu 14, and a differentially expressed eDNA library was constructed using the eDNA of Nongdafu 14 storage roots as tester and that of Kokei No. 14 storage roots as driver. Out of the 1 530 clones sequenced, we identified 292 nonredundant ESTs. GO and KEGG analyses of these differentially expressed ESTs indicated that diverse metabolism pathways were affected and candidate genes involved in regulation of carotenoids synthesis are suggested.

关键词： lpomoea batatas (L.) Lam. carotenoids suppression subtractive hybridization differentially expressed ESTs

Identifying changes in the wheat kernel proteome under heat stress using iTRAQ

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The crop Journal 2018年第6期6卷 600-610页

作者： Yufeng Zhang Hongyao Lou Dandan Guo Ruiqi Zhang Meng Su Zhenghong Hou Haiying Zhou Rongqi Liang Chaojie Xie Mingshan You Baoyun Li beijing key laboratory of crop genetic improvement/key laboratory of crop heterosis & utilization College of Agronomy and BiotechnologyChina Agricultural University

Wheat(Triticum aestivum L.) is one of the three major global food crops. Hightemperature stress can affect its yield and quality. Studies of the effect of hightemperature stress on wheat kernel development are important because they can reveal the stability of wheat quality and lead to the genetic improvement of wheat quality traits. In this study, the isobaric tags for relative and absolute quantitation(iTRAQ)method was adopted to analyze changes in the protein expression profile of wheat cultivars under high temperature stress. The protein content of wheat grain increased under heat stress, while the SDS-sedimentation value and starch content *** filling was deficient under high temperature stress, which reduced thousandkernel weight but did not affect wheat kernel length. The 207 differentially expressed proteins identified in Gaocheng 8901 under heat stress were associated with energy metabolism, growth and development, and stress response. Gene Ontology enrichment analysis showed that the annotated proteins that were differentially expressed in Gaocheng 8901 under heat stress were involved mainly in stimulus response, abiotic stress response, stress response, and plasma membrane. A set of 78 differentially expressed proteins were assigned to 83 KEGG signaling/metabolic pathways. KEGG pathway enrichment analysis showed that this set of proteins was significantly enriched in members of 51 pathways, and the proteins participated mainly in protein synthesis in the endoplasmic reticulum, starch and sucrose metabolism, and reaction on ribosomes. Five differentially expressed proteins were involved in protein–protein interaction networks that may greatly influence the yield and quality of wheat grain. In wheat, high-temperature stress leads to a variety of effects on protein expression and may ultimately cause changes in yield and quality.

关键词： Winter wheat Heat stress GO enrichment KEGG pathway enrichment Interaction networks

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.

Overexpression of SOS Genes Enhanced Salt Tolerance in Sweetpotato

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Journal of Integrative Agriculture 2012年第3期11卷 378-386页

作者： GAO Shang YUAN Li ZHAI Hong LIU Cheng-long HE Shao-zhen LIU Qing-chang beijing key laboratory of crop genetic improvement/laboratory of crop heterosis and utilization Ministry of Education/College of Agriculture and BiotechnologyChina

The production of transgenic sweetpotato （*** 18） plants exhibiting enhanced salt tolerance using salt overly sensitive （SOS） genes was achieved through Agrobacterium tumefaciens-mediated *** strain EHA105 harbors a binary vector pCAMBIA3301 with SOS genes （SOS1,SOS2 and SOS3） and bar *** culture was conducted using 0.3 mg L^-1 phosphinothricin （PPT）.A total of 40 plants were produced from the inoculated 170 cell aggregates via somatic *** analysis showed that 37 of the 40 regenerated plants were transgenic *** in vitro assay demonstrated that superoxide dismutase （SOD） and proline were significantly more accumulated and malonaldehyde （MDA） was significantly less accumulated in 21 transgenic plants than in control plants when they were exposed to 86 mmol L^-1 *** tolerance of these 21 plants was further evaluated with Hoagland solution containing 0,51,86,and 120 mmol L^-1 NaCl in the *** results indicated that 6 of them had significantly better growth and rooting ability than the remaining 15 transgenic plants and control *** of SOS genes in the 6 salt-tolerant transgenic plants was demonstrated by RT-PCR *** study provides an alternative approach for improving salt tolerance of sweetpotato.

关键词： Agrobacterium tumefaciens sweetpotato salt tolerance SOS transgenic plant

Proteomic Analysis of Wheat Seed in Response to Drought Stress

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Journal of Integrative Agriculture 2014年第5期13卷 919-925页

作者： ZHANG Yu-feng HUANG Xiu-wen WANG Li-li WEI Liu WU Zhi-hui YOU Ming-shan LI Bao-yun beijing key laboratory of crop genetic improvement/key laboratory of crop heterosis & utilization Ministry of Education/College of Agronomy and Biotechnology China Agricultural University

Drought stress is one of the major factors affecting in wheat yield and grain quality. In order to investigate how drought stress might influence wheat quality during grain filling, three wheat cultivars Gaocheng 8901, Jagger and Nongda 3406 were subjected to drought stress during the grain filling stage. Neither globulin and glutenin, nor the relative percentage ofamylose significantly changed following drought treatments, whereas albumin and gliadin concentrations did. The SDS-sedimentation, which has a strong linear correlation with wheat baking quality was markedly decreased following drought stress. These results indicated that drought had an adverse effect on wheat qualit3,. In order to investigate the protein complexes in the wheat flour, the data from native PAGE and SDS-PAGE were combined and a total of 14 spots were successfully identified, and of these eight protein types were determined to be potential complex forming proteins.

关键词： wheat seed drought stress two-dimensional electrophoresis (2-DE)

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