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Competitive Expression of Endogenous Wheat CENH3 May Lead to Suppression of Alien ZmCENH3 in Transgenic Wheat × maize Hybrids

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Journal of Genetics and Genomics 2015年第11期42卷 639-649页

作者： Wei Chen Qilin Zhu Haiyan Wang Jin Xiao Liping Xing Peidu Chen Weiwei Jin Xiu-E.Wang State Key Laboratory of Crop Genetics and Germplasm Enhancement Cytogenetics InstituteNanjing Agricultural University/JCIC-MCP national maize improvmental center China Agricultural University

Uniparental chromosome elimination in wheat × maize hybrid embryos is widely used in double haploid production of wheat. Several explanations have been proposed for this phenomenon, one of which is that the lack of cross-species CENH3 incorporation may act as a barrier to interspecies hybridization. However, it is unknown if this mechanism applies universally. To study the role of CENH3 in maize chromosome elimination of wheat x maize hybrid embryos,？cos, maize ZmCENH3 and wheat aTaCENH3-B driven by the constitutive CaMV35S promoter were transformed into wheat variety Yangmai 158. Five transgenic lines for ZmCENH3 and six transgenic lines for ctTaCENH3-B were identified. RT-PCP analysis showed that the transgene could be transcribed at a low level in all ZmCENH3 transgenic lines, whereas transcription of endogenous wheat CENH3 was significantly up-regulated. Interestingly, the expression levels of both wheat CENH3 and ZmCENH3 in the ZmCENH3 transgenic wheat × maize hybrid embryos were higher than those in the non-transformed Yangmai 158 × maize hybrid embryos. This indicates that the alien ZmCENH3 in wheat may induce competitive expression of endogenous wheat CENH3, leading to suppression of ZmCENH3 over-expression. Eliminations of maize chromosomes in hybrid embryos of ZmCENH3 transgenic wheat ×maize and Yangmai 158 x maize were compared by observations on micronuelei presence, by marker analysis using maize SSRs （simple sequence repeats）, and by FISH （fluorescence in situ hybridization） using 45S rDNA as a probe. The results indicate that maize chromosome elimination events in the two crosses are not sigmficantly different. Fusion protein ZmCENH3- YFP could not be detected in ZmCENH3 transgenic wheat by either Western blotting or immnunostaining, whereas accumulation and loading of the αTaCENH3-B-GFP fusion protein was normal in aTaCENH3-B transgenic lines. As ZmCENH3-YFP did not accumulate after AM114 treatment, we speculate that low levels of ZmCENH3 protein in

关键词： CENH3 Transgenic wheat Uniparental chromosome elimation Triticum aestivura Zea mays

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Decrease in DNA methylation 1(DDM1) is required for the formation of ~mCHH islands in maize

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Journal of Integrative Plant Biology 2019年第6期61卷 749-764页

作者： Jin Cheng Long Ai Ai Xia Jing Han Liu Ju Li Jing Ya Zhong Wang Chuang Ye Qi Yan He national maize Improvement center of China China Agricultural University

DNA methylation plays a crucial role in suppressing mobilization of transposable elements and regulation of gene expression. A number of studies have indicated that DNA methylation pathways and patterns exhibit distinct properties in different species, including Arabidopsis, rice, and ***, we characterized the function of DDM1 in regulating genome-wide DNA methylation in maize. Two homologs of Zm DDM1 are abundantly expressed in the embryo and their simultaneous disruption caused embryo lethality with abnormalities in cell proliferation from the early stage of kernel development. We establish that Zm DDM1 is critical for DNA methylation, at CHG sites, and to a lesser extent at CG sites, inheterochromatic regions, and unexpectedly, it is required for the formation ofmCHH islands. In addition, Zm DDM1 is indispensable for the presence of 24-nt si RNA, suggesting its involvement in the Rd DM pathway. Our results provide novel insight into the role of Zm DDM1 in regulating the formation of mCHH islands, via the Rd DM pathway maize, suggesting that,in comparison to Arabidopsis, maize may have adopted distinct mechanisms for regulating ~mCHH.

关键词： Decrease in DNA methylation suppressing mobilization pathway maize

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Fosmid library construction and screening for the maize mutant gene Vestigial glume 1

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The Crop Journal 2016年第1期4卷 55-60页

作者： Chaoxian Liu Xiaoli Liu Lei Lei Haiying Guan Yilin Cai maize Research Institute Southwest University national maize Improvement center of China China Agricultural University maize Institute Shandong Academy of Agricultural Sciences

The maize mutant gene Vestigial glume 1(Vg1) has been fine-mapped to a narrow region by map-based cloning and the candidate gene for Vg1 spanned 19.5 kb. Here we report Vg1 genomic fosmid library construction and screening. The fosmid library of Vg1 consisted of574,000 clones with an average insert size of 36.4 kb, representing 7.9-fold coverage of the maize genome. Fosmid stability assays indicated that clones were stable during propagation in the fosmid system. Using Vg1 candidate gene-specific primers, a positive clone was successfully identified. This discovery will pave the way for identifying the function of Vg1 in maize development.

关键词： Vestigial glume 1 Fosmid Library construction Library screening

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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

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CCT family genes in cereal crops: A current overview

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The Crop Journal 2017年第6期5卷 449-458页

作者： Yipu Li Mingliang Xu national maize Improvement center of China China Agricultural University Beijing 100193 China

Control of flowering time is crucial for reproductive success of cereal crops, and has a significant impact on grain yield as well as adaptation to diverse environmental *** integrate signals from both environmental cues and endogenous regulatory pathways to fine-tune flowering time. The CCT domain originally described to a 43-amino acid sequence at the C-terminus of three Arabidopsis proteins, namely CONSTANS(CO),CO-LIKE, and TIMING OF CAB1(TOC1). The CCT domain-containing genes(CCT genes),which encode transcription co-factors, are the major genetic determinants that modulate flowering time, and this in turn enables plants to effectively expand their territory to take advantage of favorable habitats. Moreover, certain CCT genes have pleiotropic effects on morphological traits and confer resistance/tolerance to biotic/abiotic stresses. CCT genes can be classified into three families, namely COL(CONSTANS-like), PRR(Pseudo-response regulator), and CMF(CCT motif family),based on their non-CCT domains. During domestication, natural and artificial selection resulted in reduced nucleotide diversity of CCT genes in modern cultivated cereals than their wild types. Here, we review the features and functions of CCT genes in cereal crops and propose future research to focus on CCT genes and their utilization in crop breeding.

关键词： CCT domain Flowering time Photoperiod Pleiotropy

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A Sequential Quantitative Trait Locus Fine-Mapping Strategy Using Recombinant-Derived Progeny

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Journal of Integrative Plant Biology 2012年第4期54卷 228-237页

作者： Qin Yang Dongfeng Zhang Mingliang Xu national maize Improvement center of China China Agricultural UniversityBeijing 100193China

A thorough understanding of the quantitative trait loci （QTLs） that underlie agronomically important traits in crops would greatly increase agricultural productivity. Although advances have been made in QTL cloning, the majority of QTLs remain unknown because of their low heritability and minor contributions to phenotypic performance. Here we summarize the key advantages and disad- vantages of current QTL fine-mapping methodologies, and then introduce a sequential QTL fine-mapping strategy based on both genotypes and phenotypes of progeny derived from recombinants. With this mapping strategy, experimental errors could be dramat- ically diminished so as to reveal the authentic genetic effect of target QTLs. The number of progeny required to detect QTLs atvarious R2 values was calculated, and the backcross generation suitable to start QTL fine-mapping was also estimated. This mapping strategy has proved to be very powerful in narrowing down QTL regions, particularly minor-effect QTLs, as revealed by fine-mapping of various resistance QTLs in maize. Application of this sequential QTL mapping strategy should accelerate cloning of agronomically important QTLs, which is currently a substantial challenge in crops.

关键词： Fine-mapping strategy progeny quantitative trait locus recombinant.

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Signal Peptide of Potato PinII Enhances the Expression of Cry1Ac in Transgenic Tobacco

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Acta Biochimica et Biophysica Sinica 2004年第8期36卷 553-558页

作者： Yun-Jun LIU, Yuan YUAN, Jun ZHENG, Ya-Zhong TAO, Zhi-Gang DONG,Jian-Hua WANG, and Guo-Ying WANGState Key Laboratory for Agrobiotechnology and national center for maize Improvement, China Agricultural University, Beijing 100094, China State Key Laboratory for Agrobiotechnology and national center for maize Improvement China Agricultural UniversityBeijing 100094 China

The modified Cry1Ac was expressed in transgenic tobacco plants. To allow secretion of the Cry1Ac protein into the intercellular space, the signal peptide sequence of potato proteinase inhibitor II (pinII) was N-terminally fused to the Cry1Ac encoding region. Expression of Cry1Ac in transgenic tobacco plants was assayed with ELISA. The results showed that pinII signal peptide sequence enhanced the expression of Cry1Ac protein and led to the secretion of the Cry1Ac protein in transgenic tobacco plants. GFP gene was also fused to the signal peptide sequence and transformed to tobacco. The results of fluorescent detection showed that GFP had localized in the apoplast of transgenic plants.

关键词： Cry1Ac signal peptide GFP transgenic tobacco

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Identification and fine mapping of quantitative trait loci for the number of vascular bundle in maize stem

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Journal of Integrative Plant Biology 2016年第1期58卷 81-90页

作者： Cheng Huang Qiuyue Chen Guanghui Xu Dingyi Xu Jinge Tian Feng Tian Department of Plant Genetics and Breeding National Maize Improvement Center of China China Agricultural University

Studies that investigated the genetic basis of source and sink related traits have been widely ***, the vascular system that links source and sink received much less attention. When maize was domesticated from its wild ancestor, teosinte, the external morphology has changed dramatically; however, less is known for the internal anatomy changes. In this study, using a large maize-teosinte experimental population, we performed a high-resolution quantitative trait locus（QTL） mapping for the number of vascular bundle in the uppermost internode of maize *** results showed that vascular bundle number is dominated by a large number of small-effect QTLs, in which a total of 16 QTLs that jointly accounts for 52.2% of phenotypic variation were detected, with no single QTL explaining more than 6% of variation. Different from QTLs for typical domestication traits, QTLs for vascular bundle number might not be under directional selection following *** Near Isogenic Lines（NILs） developed from heterogeneous inbred family（HIF）, we further validated the effect of one QTL qVb9-2 on chromosome 9 and fine mapped the QTL to a 1.8-Mb physical region. This study provides important insights for the genetic architecture of vascular bundle number in maize stem and sets basis for cloning of qVb9-2.

关键词： Fine-mapping genetic architecture maize quantitativetrait locus vascular bundle number

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Relationship between differ- ential gene expression pat- terns in functional leaves of maize (Zea mays L.) at milk filling stage and heterosis using cDNA-AFLP

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Chinese Science Bulletin 2003年第1期48卷 76-81页

作者： TIAN Zengyuan & DAI Jingrui national maize Improvement center of China, China Agricultural Uni-versity, Beijing 100094, China Correspondence should be addressed to Tian Zengyuan (e-mail: tian-zengyuan@***) national maize Improvement center of China China Agricultural University Beijing China

To understand the molecular mechanism of maize heterosis, differential gene expression patterns in the functional leaves of 35 maize hybrids relative to their parents involving 10 elite inbreds at milk filling stage were analyzed by using cDNA-AFLP. The correlation analyses of various differential expression patterns with the performance and heterosis of main maize agronomic traits were evaluated. The main results were as follows: For uniparental specific expression, significant positive correlations were detected with the performance of seed weight per ear and 100-seed weight at 0.01 and 0.05 probability levels respectively. For biparental specific expression, significant negative correla-tions were detected with the performance of ear diameter and seed weight per ear at 0.01 probability level. For uni-parental specific expression, significant positive correlations were detected with the heterosis of ear diameter and seed weight per ear at 0.01 and 0.05 probability levels respectively. For biparental specific expression, significant negative cor-relation was detected with the heterosis of ear diameter at 0.05 probability level. However, for F1-specific expression, for fragments detected only in one parent and F1, and for fragments detected only in two parents or only in F1, no sig-nificant correlation was detected with the performance or heterosis of all agronomic traits surveyed.

关键词：玉米灌浆期叶片差异基因表达模式杂种优势相关性 cDNA-AFLP 农艺性状

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Selection and development of representative simple sequence repeat primers and multiplex SSR sets for high throughput automated genotyping in maize

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Chinese Science Bulletin 2007年第2期52卷 215-223页

作者： WANG FengGe ZHAO JiuRan DAI JingRui YI HongMei KUANG Meng SUN YanMei YU XinYan GUO JingLun WANG Lu national maize Improvement center of China China Agriculture University Beijing 100094 China maize Research center Beijing Academy of Agricultural & Forestry Sciences Beijing 100097 China

In the current study, 1900 maize simple sequence repeat (SSR) primers published in maizeGDB were screened utilizing reference literature, 15 representative Chinese maize inbred lines and 15 Chinese maize hybrids from national regional testing. In total, 500 highly polymorphic primers were identified and used to construct a genetic map. 100 evenly distributed primers, 10 primers per chromosome, were further selected as a set of universal SSR core primers, recommended as preferred primers for general studies. These core primers were then redesigned and used to construct a high throughput multiplex PCR system based on a five-color fluorescence capillary detection system. We report here that two sets of ten-plex PCR combinations have been constructed, each consisting of 10 primers, with one primer per chromosome.

关键词：基因序列玉米分子标记遗传型

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