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Integrated transcriptomics and metabolomics analyses provide insights into cold stress response in wheat

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The crop Journal 2019年第6期7卷 857-866页

作者： Yong Zhao Meng Zhou Ke Xu Jiahao Li Shanshan Li Shuhua Zhang Xueju Yang huabei key laboratory of crop germplasm Hebei Agricultural UniversityBaoding 071000HebeiChina College of Life Sciences Hebei UniversityBaoding 071000HebeiChina

Cold tolerance of crop plants influences survival and productivity under low-temperature conditions. Elucidation of molecular mechanisms underlying low temperature tolerance could be helpful in breeding. In this study, we used integrated transcriptomics and metabolomics analyses to investigate changes in gene/metabolite activity in a winter-hardy wheat cultivar of(cv. Jing 411) when subjected to sold stress. The 223 metabolites mainly enriched during cold acclimation included carbohydrates, flavonoids, and amino *** common metabolites had altered abundance following freezing treatment;six increased and two decreased. Transcriptome analysis revealed that 29,066 genes were differentially expressed in wheat crowns after cold acclimation compared to the nonacclimated control. Among them, 745 genes were up-regulated following freezing treatment, suggesting substantial change in expression of a large quantity of genes upon cold acclimation and freezing treatment, which impacts on the modified *** analysis of gene expression and metabolite profiles revealed that the abscisic acid(ABA)/jasmonic acid(JA) phytohormone signaling and proline biosynthesis pathways were significantly modulated under cold acclimation and freezing treatments. Our results indicated that low-temperature stress induced substantial changes in both transcriptomes and metabolomes. Critical pathways associated with ABA/JA signaling and proline biosynthesis played important roles in regulating cold tolerance in wheat.

关键词： Cold tolerance Transcriptome Metabolome Abscisic acid Jasmonic acid Signaling pathways

Identification and Preliminary Analysis of Several Centromere-associated Bacterial Artificial Chromosome Clones from a Diploid Wheat Library

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Journal of Integrative Plant Biology 2006年第3期48卷 348-358页

作者： Zhao Liu Wei Yue Yu-Shen Dong Xue-Yong Zhang key laboratory of crop germplasm and Biotechnology Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China State key laboratory of crop Genetics and germplasm Enhancement Nanjing Agricultural University Nanjing 210095 China

Although the centromeres of some plants have been investlgated prevlously, our knowledge of the wheat centromere Is still very llmlted. To understand the structure and functlon of the wheat centromere, we used two centromeric repeats （RCS1 and CCS1-5ab） to obtain some centromere-assoclated bacterial artificial chromosome （BAC） clones in 32 RCS1-related BAC clones that had been screened out from a diploid wheat （Triticum boeoticum Boiss.; 2n=2x=14） BAC library. Southern hybridization results indicated that, of the 32 candidates, there were 28 RCS1-positive clones. Based on gel blot patterns, the frequency of RCS1 was approximately one copy every 69.4 kb in these 28 RCS1-positive BAC clones. More bands were detected when the same filter was probed with CCS1-5ab. Furthermore, the CCS1 bands covered all the bands detected by RCS1, which suggests that some CCS1 repeats were distributed together with RCS1. The frequency of CCS1 families was once every 35.8 kb, nearly twice that of RCS1. Fluorescence in situ hybridization （FISH） analysis Indicated that the five BAC clones containing RCS1 and CCS1 sequences all detected signals at the centromerlc regions in hexaplold wheat, but the signal intensities on the A-genome chromosomes were stronger than those on the B- and/or Dgenome chromosomes. The FISH analysis among nine Triticeae cereals indicated that there were A-genomespecific （or rich） sequences dispersing on chromosome arms in the BAC clone TbBACS. In addition, at the interphase cells, the centromeres of diploid species usually clustered at one pole and formed a ring-like allocation In the period before metaphase.

关键词： centromere fluorescence in situ hybridization (FISH) repeat Southern hybridization Triticum boeoticum.

Development of Heat Tolerant Two-Line Hybrid Rice Restorer Line Carrying Dominant Locus of OsHTAS

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Rice science 2021年第1期28卷 99-108页

作者： Mehmood Jan Gulmeena Shah Huang Yuqing Liu Xuejiao Zheng Peng Du Hao Chen Hao Tu Jumin Zhejiang key laboratory of crop germplasm Resources Institute of Crop ScienceZhejiang UniversityHangzhou 310058ZhejiangChina

In order to create novel germplasm resources for breeding heat tolerant variety,we transferred a dominant allele OsHTAS,previously characterized and cloned from a high-temperature tolerant local variety HT54,which was collected from the rice production area of southern China,into a high-temperature sensitive intermediate breeding line HT13 through six rounds of successive backcross by using marker-assisted *** molecular analysis showed that the recovery of genetic background of a resultant near isogenic line(NIL),MHT13,was around 99.8%.The OsHTAS gene introduced in the MHT13 expressed normally in the HT13 genetic background,mediating heat tolerance and phenotype similar to those of the donor parent *** major agronomic traits of MHT13 resembled those of the recurrent parent ***,MHT13 had high general combining ability and its rice quality reached the grade 3 standard of edible high-quality rice issued by Ministry of Agriculture of the People’s Republic of China,which greatly improved its application value in rice production.

关键词： rice heat tolerance high temperature yield near isogenic line marker-assisted selection grain quality OsHAT gene

The G-protein α subunit GhGPA positively regulates Gossypium hirsutum resistance to Verticillium dahliae via induction of SA and JA signaling pathways and ROS accumulation

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The crop Journal 2021年第4期9卷 823-833页

作者： Bin Chen Yan Zhang Jun Yang Man Zhang Qingming Ma Xingfen Wang Zhiying Ma State key laboratory of North China crop Improvement and Regulation/key laboratory for crop germplasm Resources of Hebei Hebei Agricultural UniversityBaoding 071001HebeiChina

Verticillium wilt, a devastating disease in cotton caused by Verticillium dahliae, reduces cotton quality and yield. Heterotrimeric GTP-binding proteins, consisting of Ga, Gb, and Gc subunits, transducers of receptor signaling, function in a wide range of biological events. However, the function of Ga proteins in the regulation of defense responses in plants is largely unexplored, except for a few reports on model species. In the present study, a cotton G-protein a-subunit-encoding gene(GhGPA) was isolated from Verticillium wilt-resistant Gossypium hirsutum(upland cotton) cv. ND601. GhGPA transcription was up-regulated under V. dahliae stress, with higher expression in tolerant than in susceptible cotton *** localization revealed GhGPA to be located in the plasma membrane. GhGPA shows high(85.0%) identity with Arabidopsis AT2 G26300(AtGPA1), and AtGPA1 gpa1-4 mutants displayed susceptibility to V. dahliae. Ectopic expression of GhGPA successfully restored the resistance of Arabidopsis gpa1-4 mutants to Verticillium wilt and made them more resistant than the wild type. Overexpression of GhGPA in Arabidopsis markedly increased the resistance and resulted in dramatic up-regulation of pathogenesis-related(PR) genes and increased in H2 O2 accumulation and salicylic acid(SA) and jasmonic acid(JA) contents. However, suppressing GhGPA expression via virus-induced gene silencing(VIGS)increased susceptibility to Verticillium wilt, down-regulated the expression of PR and marker genes in SA and JA signaling pathways, and reduced H2 O2 content. The contents of SA and JA in Arabidopsis gpa1-4 and VIGS cotton were lower than those in the wild type and empty-vector control. However,GhGPA-overexpressing Arabidopsis contained more SA and JA than the wild type when inoculated with V. dahliae. Thus, GhGPA plays a vital role in Verticillium wilt resistance by inducing SA and JA signaling pathways and regulating the production of reactive oxygen species. These findings

关键词： Gossypium hirsutum GhGPA Verticillium wilt Pathogenesis-related genes ROS

Molecular Cloning,Expression Analysis and Localization of Exo70A1 Related to Self Incompatibility in Non-Heading Chinese Cabbage(Brassica campestris ***)

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Journal of Integrative Agriculture 2013年第12期12卷 2149-2156页

作者： WANG Li GE Ting-ting PENG Hai-tao WANG Cheng LIU Tong-kun HOU Xi-lin LI Ying Horticultural Department Nanjing Agricultural University State key laboratory of crop Genetics and germplasm Enhancement key laboratory of Southern Vegetable crop Genetic Improvement Ministry of Agriculture

The exocyst is a conserved protein complex,and required for vesicles tethering,fusion and polarized ***70A1,the exocyst subunit,is essential for assembly of the exocyst *** better understand potential roles of Exo70A1 in non-heading Chinese cabbage（Brassica campestris ***）,we obtained the full-length cDNA of Exo70A1 gene,which consisted of 1 917 bp and encoded a protein of 638 amino *** showed BcExo70A1 shared 94.9% identity with Brassica oleracea ***（AEI26267.1）,and clustered into a same group with other homologues in *** *** and Brassica *** localization analysis showed BcExo70A1 was localized to punctate structures in cytosol of onion epithelial *** showed that BcExo70A1 was widely presented in stamens,young stems,petals,unpollinated pistils,roots and leaves of self compatible and incompatible *** transcripts of BcExo70A1 in non- heading Chinese cabbage declined during initial 1.5 h after incompatible pollination,while an opposite trend was presented after compatible *** study reveals that BcExo70A1 could play essential roles in plant growth and development,and is related to the rejection of self pollen in non-heading Chinese cabbage.

关键词： Exo70A1 gene expression subcellular localization self-incompatibility non-heading Chinese cabbage

Detection of Tocopherol in Oilseed Rape (Brassica napus L.) Using Gas Chromatography with Flame Ionization Detector

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Journal of Integrative Agriculture 2013年第5期12卷 803-814页

作者： Nazim Hussain Zahra Jabeen LI Yuan-long CHEN Ming-xun LI Zhi-lan GUO Wan-li Imran Haider Shamsi CHEN Xiao-yang JIANG Li-xi key laboratory of crop germplasm Resources of Zhejiang Province College of Agriculture and BiotechnologyZhejiang University

The variation among Chinese genotypes of Brassica napus L. for seed tocopherols content and their analysis using gas chromatography has not been comprehensively reported till to date. In the present study, the tocopherol contents of four Chinese genotypes of Brassica napus L., namely, Gaoyou 605, Zhejiang 619, Zheshuang 758, and Zheshuang 72, were evaluated using three modified sample preparation protocols （P1, P2, and P3） for tocopherol extraction. These methods were distinguished as follows. Protocol one （P1） included the evaporation of solvent after extraction without silylation. Protocol two （P2） followed the direct supernatant collection after overnight extraction without drying and silylation. Protocol three （P3） included trimethylsilylation with N,O-bis（trimethylsilyl） trifluoroacetamide. Genotypic comparison of tocopherol and its isoforms revealed that Gaoyou 605 was dominant over the other genotypes with （140.5＋ 10.5）, （316.2＋ 9.2）, and （559.1＋ 24.3） ~tg g-~ of seed meal ct-, 7-, and total （T-） tocopherol, respectively, and a 0.44＋0.04 ^- to 7-tocopherol ratio. The comparison of the sample preparation protocols, on the other hand, suggests that P3 is the most suitable method for the tocopherol extraction from Brassica oilseeds and for the analysis of tocopherols using gas chromatography flame ionization detector （GC-FID）. Trimethylsilylation is the key step differentiating P3 from P1 and P2. Variations detected in tocopherol contents among the Chinese rapeseed （B. napus） genotypes signify the need to quantify a wide range of rapeseed germplasm for seed tocopherol dynamics in short and crop improvement in long.

关键词： Brassica napus L. genotypic variation tocopherol analysis gas chromatography flame ionization detector

Development of EST-PCR Markers for the Chromosome 4V of Haynaldia villosa and Their Application in Identification of 4V Chromosome Structural Aberrants

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Journal of Integrative Agriculture 2014年第2期13卷 282-289页

作者： ZHAO Ren-hui WANG Hai-yan JIA Qi XIAO Jin YUAN Chun-xia ZHANG Ya-jun HU Qing-shan WANG Xiu-e State key laboratory of crop Genetics and germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University

EST-PCR based molecular markers specific for alien chromosomes are not only useful for the detection of the introgressed alien chromatin in the wheat background, but also provide evidence of the syntenic relationship between homoeologous chromosomes. In the present study, in order to develop high density and evenly distributed molecular markers on chromosome 4V of Haynaldia villosa, a total of 607 primer pairs were designed according to the EST sequences, which were previously located in 23 different bins of wheat chromosomes 4A, 4B and 4D. By using the Triticum durum-H, villosa amphiploid and T. aestivum-H, villosa alien chromosome lines involving chromosome 4V, it was found that 9.23% of the tested primers could amplify specific bands for chromosome 4V. Thirty and twenty-six specific markers could be assigned to chromosome arms 4VS and 4VL, respectively. These 4V specific markers provided efficient tools for the characterization of structural variation involving the chromosome 4V as well as for the selection of useful genes located on chromosome 4V in breeding programs.

关键词： Haynaldia villosa molecular marker Triticum aestivum chromosome engineering

Quantification and analysis of anthocyanin and flavonoids compositions,and antioxidant activities in onions with three different colors

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Journal of Integrative Agriculture 2016年第9期15卷 2175-2181页

作者： ZHANG Shi-lin DENG Peng XU Yu-chao Lü Shan-wu WANG Jian-jun State key laboratory of crop Genetics and germplasm Enhancement Key Laboratory of Biology and Germplasm Enhancement of Horticultural Crops in East China Ministry of Agriculture/College of Horticulture Nanjing Agricultural University

Onion（Allium cepa L.） is a very important vegetable crop widely cultivated throughout the world. In this study, we measured total anthocyanins and flavonols of onions of three colors. The fresh weight（FW） of total anthocyanins were（29.99±1.19）,（9.64±1.30） and（0.75±0.40） mg 100 g^–1 fruit in red, yellow and white onions, respectively. Likewise, the FW of total flavonoids were（111.10±5.98）,（36.64±3.59）, and 0 mg 100 g^–1 in red, yellow and white, respectively. Four types of anthocyanins（delphinidin 3,5-diglycosides, cyanidin 3,5-diglycosides, cyanidin 3-glycosides and cyanidin 3-（6′′-malonyl）-glucopyranoside） and two kinds of flavonoids（quercetin and quercetin 3-glycosides） were identified in two varieties（red and yellow bulb） of onions by HPLC/DAD-ESI/MS. The total polyphenol contents were also measured by means of Folin-Ciocalteu method. Moreover, all the concentrations of anthocyanins, flavonoids and polyphenols showed significantly positive correlations with antioxidant activities measured by DPPH˙, ABTS˙＋ and FRAP assays. This study provided information on anthocyanin and flavonoids compositions that will be useful for onion breeding.

关键词： onion(Allium cepa L.) anthocyanins flavonoids antioxidant activity HPLC-DAD-ESI/MS

Genomics-based plant germplasm research(GPGR)

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The crop Journal 2017年第2期5卷 166-174页

作者： Jizeng Jia Hongjie Li Xueyong Zhang Zichao Li Lijuan Qiu key laboratory of crop germplasm & Biotechnology Ministry of AgricultureInstitute of Crop ScienceChinese Academy of Agricultural SciencesThe National Key Facility for Crop Gene Resources and Genetic ImprovementNFCRGI Beijing key laboratory of crop Genetic Improvement Key Laboratory of Crop Heterosis and UtilizationMinistry of EducationChina Agricultural University

Plant germplasm underpins much of crop genetic improvement. Millions of germplasm accessions have been collected and conserved ex situ and/or in situ, and the major challenge is now how to exploit and utilize this abundant resource. Genomics-based plant germplasm research （GPGR） or ＂Cenoplasmics＂ is a novel cross-disciplinary research field that seeks to apply the principles and techniques of genomics to germplasm research. We describe in this paper the concept, strategy, and approach behind GPGR, and summarize current progress in the areas of the definition and construction of core collections, enhancement of germplasm with core collections, and gene discovery from core collections. GPGR is opening a new era in germplasm research. The contribution, progress and achievements of GPGR in the future are predicted.

关键词： Plant germplasm Genomics Genomics-based plant germplasm research(GPGR) Genoplasmics