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CRISPR/Cas9 editing of wheat TaQ genes alters spike morphogenesis and grain threshability

Journal of Genetics and Genomics 2020年第9期47卷 563-575页

作者： Huiyun Liu Ke Wang Huali Tang Qiang Gong lipu du Xinwu Pei Xingguo Ye Institute of Crop Science Chinese Academy of Agricultural SciencesBeijing100081China Biotechnology Research Institute Chinese Academy of Agricultural SciencesBeijing100081China

The Ta Q alleles as one of the AP2-like transcription factors in common wheat(Triticum aestivum) play an important role in the evolution of spike characteristics from wild and domesticated emmer to modern wheat cultivars. Its loss-of-function mutant not only changed threshability and spike architecture but also affected plant height, flowering time, and floret structure. However, the comprehensive functions of Ta AQ and Ta Dq genes in wheat have not been fully elucidated yet. Here, CRISPR/Sp Cas9 was used to edit wheat Ta AQ and Ta Dq. We obtained homozygous plants in the T1 generation with loss of function of only Ta AQ or Ta Dq and simultaneous loss of function of Ta AQ and Ta Dq to analyze the effect of these genes on wheat spikes and floret shapes. The results demonstrated that the Ta AQ-edited plants and the Ta AQ and Ta Dq simultaneously-edited plants were nearly similar in spike architecture, whereas the Ta Dq-edited plants were different from the wild-type ones only in plant height. Moreover, the Ta AQ-edited plants or the Ta AQ and Ta Dq simultaneously-edited plants were more brittle than the wild-type and the Ta Dqedited plants. Based on the expression profiling, we postulated that the VRN1, FUL2, SEP2, SEP5, and SEP6 genes might affect the number of spikelets and florets per spike in wheat by regulating the expression of Ta Q. Combining the results of this report and previous reports, we conceived a regulatory network of wheat traits, including plant height, spike shape, and floral organs, which were influenced by AP2-like family genes. The results achieved in this study will help us to understand the regulating mechanisms of Ta AQ and Ta Dq alleles on wheat floral organs and inflorescence development.

关键词： Triticum aestivum Gene editing Spike architecture APETALA2 TaQ genes

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Development of a wheat material with improved bread-making quality by overexpressing HMW-GS 1S^(l)x2.3^(*)from Aegilops longissima

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The Crop Journal 2022年第6期10卷 1717-1726页

作者： Yuliang Qiu Haiqiang Chen Shuangxi Zhang Jing Wang lipu du Ke Wang Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China Crop Research Institute Ningxia Academy of Agri-Forestry SciencesYinchuan 750105NingxiaChina

Wheat bread-making quality can be improved by use of high-molecular-weight glutenin subunits(HMW-GSs)from wild *** longissima is a close relative of wheat that contains a number of HMW-GS-encoding genes including 1S^(l)x2.3^(*).In this study,transgenic wheat lines overexpressing 1S^(l)x2.3^(*)were obtained by Agrobacterium-mediated transformation and used to investigate the genetic contribution of 1S^(l)x2.3^(*)to wheat flour-processing *** 1S^(l)x2.3^(*)transgene was stably inherited and expressed over *** of 1S^(l)x2.3^(*)increased the relative expression of 1Dx2 and 1Dy12 and reduced that of 1By18 during grain *** general,integration of 1S^(l)x2.3^(*)stimulated the accumulation of endogenous HMW-GSs and low-molecular-weight glutenin subunits in wheat kernels,greatly increasing the glutenin:gliadin ratio and resulting in faster formation of protein bodies in the endosperm during grain development.A wheat material with improved flour-making quality was developed in which 1S^(l)x2.3^(*)improved wheat bread-making quality.

关键词： Wheat Aegilops longissima High-molecular-weight glutenin subunits 1S^(l)x2.3^(*) Bread-making quality

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Development and genetic analysis of wheat double substitution lines carrying Hordeum vulgare 2H and Thinopyrum intermedium 2Ai#2 chromosomes

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The Crop Journal 2019年第2期7卷 163-175页

作者： Jing Wang Chang Liu Xianrui Guo Ke Wang lipu du Zhishan Lin Xingguo Ye National Key Facility of Crop Gene Resources and Genetic Improvement Institute of Crop Sciences Chinese Academy of Agricultural Sciences State Key Laboratory of Plant Cell and Chromosome Engineering Institute of Genetics and Developmental Biology Chinese Academy of Sciences

Thinopyrum intermedium and barley are two close relatives of wheat and carry many genes that are potentially valuable for the improvement of various wheat traits. In this study we created wheat double substitution lines by hybridizing different wheat–Th. intermedium and wheat–barley disomic alien substitution lines, with the aim of using genes in Th. intermedium and barley for wheat breeding and investigating the genetic behavior of alien chromosomes and their wheat homoeologs. As expected, we obtained two types of wheat double substitution lines,2D2Ai#2(2B)2H( A) and 2A2 Ai#2(2B)2H(2D), in which different group 2 wheat chromosomes were replaced by barley chromosome 2 H and Th. intermedium chromosome 2Ai#2. The new materials were characterized using molecular markers, genomic in situ hybridization(GISH), and fluorescent in situ hybridization(FISH). GISH and FISH experiments revealed that the double substitution lines harbor 42 chromosomes including 38 wheat chromosomes, a pair of barley chromosomes, and a pair of Th. intermedium chromosomes. Analysis using specific DNA markers showed that two pairs of wheat homoeologous group 2 chromosomes in the new lines were substituted by a pair of 2H and a pair of 2Ai#2 chromosomes. Chromosome 2H showed a higher transmission rate than 2Ai#2, and both chromosomes were preferentially transmitted between generations via female gametes. Evaluation of botanic and agronomic traits demonstrated that,compared with their parents, the new lines showed similar growth habits and plant type but differences in plant height, flowering date, and self-fertility. Cytological observations using different probes suggested that the double substitution lines showed nearly normal genetic behavior before and during meiosis. The novel substitution lines can potentially be used in wheat meiosis research and breeding programs.

关键词： Wheat Double substitution lines Molecular markers Transmission rate of alien chromosomes Genomic in situ hybridization

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Development of powdery mildew resistant derivatives of wheat variety Fielder for use in genetic transformation

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The Crop Journal 2023年第2期11卷 573-583页

作者： Xiaona Liang Xiaomin Bie Yuliang Qiu Ke Wang Zujun Yang Yuqi Jia Zhiying Xu Mei Yu lipu du Zhishan Lin Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China College of Life Science Shandong Agricultural UniversityTai’an 271018ShandongChina School of Life Science and Technology University of Electronic Science and Technology of ChinaChengdu 611731SichuanChina College of Agriculture Yangtze UniversityJingzhou 434025HubeiChina National Key Facility of Crop Gene Resources and Genetic Improvement Beijing 100081China Key Laboratory of Ministry of Agriculture and Rural Affairs of China for Biology and Genetic Breeding of Triticeae Crops Beijing 100081China

Genetic transformation is widely used to improve target traits and to study gene function in ***,transformation efficiency depends on the physiological status of the recipient genotype and that is affected by several factors including powdery mildew(PM)*** widely used recipient variety Fielder is very susceptible to ***,it would be beneficial to develop PM resistant derivatives with high regeneration ability for use in genetic *** the present study PM resistant lines CB037 and Pm97033 carrying genes Pm21 and PmV,respectively,were backcrossed to Fielder with selection for PM *** lines,NT89,NT90,NT154,and WT48 with Pm21 and line FL347 with PmV were developed,identified by molecular markers and genomic in situ hybridization(GISH)or fluorescent in situ hybridization(FISH),and further subjected to detailed assessment of agronomic traits and regeneration ability following genetic transformation *** FL347,WT48,NT89 and NT154 assessed as being equal to,or superior,to Fielder in regeneration and transformation ability are recommended as suitable materials for the replacement of Fielder for wheat gene transfer and genome editing study.

关键词： Genotype improvement Plant regeneration ability Transformation efficiency Triticum aestivum

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A fast technique for visual screening of wheat haploids generated from TaMTL-edited mutants carrying anthocyanin markers

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Plant Communications 2023年第3期4卷 11-13页

作者： Huali Tang Ke Wang Shuangxi Zhang Zhiyang Han Yanan Chang Yuliang Qiu Mei Yu lipu du Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China Crop Research Institute Ningxia Academy of Agriculture and Forestry SciencesYinchuan 750002China College of Agronomy Ningxia UniversityYinchuan 750021China

Dear Editor,Breeding a new wheat variety using traditional methods typically takes at least 8 to 10 years,and the breeding period can be dramatically shortened via a doubled haploid strategy,which can yield homozygotes within one or two *** the past several decades,wheat haploids have been widely induced through anther or microspore culture and chromosome elimination via interspecific hybridization between wheat and *** first technique exhibits strong genotype dependency and a severe albino phenomenon,whereas the latter shows low induction efficiency(Sangam et al.,2015).In addition,haploid induction procedures using the aforementioned methods are complicated to perform and require specialized equipment and environmentally controlled ***,the application of these technologies in wheat breeding has been limited(Sangam et al.,2015).

关键词： breeding wheat doubled

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Development of a haploid inducer by editing HvMTL in barley

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Journal of Genetics and Genomics 2023年第5期50卷 366-369页

作者： Huali Tang Yuliang Qiu Wanxin Wang Mei Yu Yanan Chang Zhiyang Han lipu du Zhishan Lin Ke Wang Xingguo Ye Institute of Crop Sciences Chinese Academy of Agricultural SciencesBeijing 100081China College of Agronomy Ningxia UniversityYinchuanNingxia 750105China

The doubled-haploid technique mediated by parthenogenesis and androgenesis in plants can directly generate homozygous diploid lines after chromosome doubling with colchicine in one or two generations,and this method undoubtedly shortens the breeding process and improves breeding efficiency in ***,haploid plants were mainly induced by anther culture or microspore culture via androgenesis(Ohnoutkova et al.,2019).In wheat(Triticum aestivum),haploids can also be induced by maize(Zea mays)pollen and chromosome elimination via parthenogenesis(Liu et al.,2020b).However,the aforementioned induction methods are genotype-dependent,require complex manipulations,and are both time-consuming and ***,it is necessary to develop new techniques or germplasm for simple and efficient haploid induction.

关键词： breeding culture doubt

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Improvement of wheat drought and salt tolerance by expression of a stress- inducible transcription factorGmDREB of soybean (Glycinemax)

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Chinese Science Bulletin 2005年第23期50卷 2714-2723页

作者： GAO Shiqing XU Huijun CHENG Xianguo CHEN Ming XU Zhaoshi LI Liancheng YE Xingguo du lipu HAO Xiaoyan MA Youzhi Key Laboratory of Crop Genetics and Breeding Ministry of Agriculture/Institute of Crop Science Chinese Academy of Agricultural Sciences Beijing 100081 China Ministry of Agriculture Key Laboratory of Plant Nutrition and Nutrient Cycling Institute of Agricultaral Resources and Regional Planning Chinese Academy of Agricultural Sciences Beijing 100081 China

Under stress conditions such as droughthigh-salinity and low-temperature, the transcription factorof DREB (dehydration responsive element binding proteins)improved efficiently stress resistance by regulating the ex-pression of its downstream genes with various environmentastress resistance in plants. GmDREB gene (GenBank Acces-sion No. AF514908) encoding a stress-inducible transcriptionfactor was cloned by screening a cDNA library of Glycinemax cv. Jinong 27 with yeast one-hybrid method. GmDREBgene was 910 bp in length and encoded 174 amino acids con-taining a conserved AP2/EREBP DNA-binding domain of 58amino acids. Two conserved functional amino acids, valineand glutamic acid, were located on the 14th and the 19thamino acid residues in the conserved structural domain. Analkaline amino acid region (KKR) related to a nuclear local-ization signal was at the N-terminal, while an acidic aminoacid region (DDD) related to trans-activation was at theC-terminal. Plant expression vectors were constructed andtransformed into wheat by bombardment. In total, 13 trans-genic plants with Ubi::GmDREB and 11 transgenic plantswith rd29A::GmDREB were identified from 103 regenerationplants by molecular analysis. The drought and salt tolerancesof T1 transgenic lines with Ubi::GmDREB orrd29A::GmDREB were demonstrated to be improved ascompared to wild type. The result also suggested that bothUbiquitin and rd29A promoters could effectively drive theexpression of the GmDREB gene and enhance drought andsalt tolerance of T1 plants.

关键词：小麦 GmDREB基因抗旱性耐盐性应力诱发转录因子大豆甘氨酸

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相敏光时域反射仪土壤振动传播特性

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激光与光电子学进展 2023年第19期60卷 77-86页

作者：王鸣封皓沙洲杜立普天津大学精密仪器与光电子工程学院精密测试技术及仪器国家重点实验室天津300072 宁夏回族自治区水利工程建设中心宁夏银川750004

相敏光时域反射仪(φ-OTDR)广泛应用于管道安全预警、光缆断点查找、清管器定位跟踪等场景,其传感光缆多埋于地下,土壤性质、传输距离、光缆结构等均对信号特征有所影响。本文研究了振动信号在土壤中的传播特性,探究了光缆结构、土壤湿... 详细信息

相敏光时域反射仪(φ-OTDR)广泛应用于管道安全预警、光缆断点查找、清管器定位跟踪等场景,其传感光缆多埋于地下,土壤性质、传输距离、光缆结构等均对信号特征有所影响。本文研究了振动信号在土壤中的传播特性,探究了光缆结构、土壤湿度、传输距离等非事件因素对系统信号特征的影响。此外,从特征频段能量变化趋势的角度,研究了真实冲击振动与非事件性信号波动的区别,并由此提出了一种外部冲击振动的判别方法。

关键词：光纤光学与光通信分布式光纤传感土壤振动传播特性特殊频段频域判断法

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基于LSTM-CNN的φ-OTDR模式识别

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光学学报 2023年第5期43卷 11-22页

作者：王鸣沙洲封皓杜立普齐敦哲天津大学精密仪器与光电子工程学院精密测试技术及仪器国家重点实验室天津300072 宁夏回族自治区水利工程建设中心宁夏银川750004

提出了以长短期记忆网络(LSTM)与卷积神经网络(CNN)为主要框架的深度学习网络,基于该网络实现模式识别。LSTM-CNN以时域曲线及其短时傅里叶变换(STFT)结果作为网络输入,LSTM提取输入信号的时序特征,CNN提取时域曲线的轮廓特征及能量特... 详细信息

提出了以长短期记忆网络(LSTM)与卷积神经网络(CNN)为主要框架的深度学习网络,基于该网络实现模式识别。LSTM-CNN以时域曲线及其短时傅里叶变换(STFT)结果作为网络输入,LSTM提取输入信号的时序特征,CNN提取时域曲线的轮廓特征及能量特征。实验中以相敏光时域反射仪(φ-OTDR)为传感系统完成数据采集。将LSTMCNN与传统的人工神经网络(ANN)及CNN对比,LSTM-CNN在各项评价指标中均处于最优状态,实现了φ-OTDR模式识别的既定目标,为实际的工程产品开发提供了概念证明与演示示例。

关键词：光通信模式识别长短期记忆网络卷积神经网络光纤预警

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基于地电流的重力流排水管道泄漏检测

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仪器仪表学报 2023年第1期44卷 154-162页

作者：杨东馥封皓齐敦哲杜立普沙洲天津大学精密测试技术及仪器国家重点实验室天津300072 宁夏回族自治区水利工程建设中心银川750002

重力流排水管道的泄漏会带来诸多不良后果,但由于重力流排水管道的特殊性,导致许多现有的管道泄漏检测方法难以适用。针对这一问题,提出了一种基于地电流的检测方法,该方法利用泄漏孔联通管道内外,通过测量在管道内移动的电极和固定在... 详细信息

重力流排水管道的泄漏会带来诸多不良后果,但由于重力流排水管道的特殊性,导致许多现有的管道泄漏检测方法难以适用。针对这一问题,提出了一种基于地电流的检测方法,该方法利用泄漏孔联通管道内外,通过测量在管道内移动的电极和固定在地面电极之间电流来确定泄漏,阐述了该方法的基本工作原理,基于该方法设计了检测系统,搭建了实验平台测试了检测效果。检测10 m长的埋地管道时,最大定位误差约0.4 m,表明该检测系统能较准确地定位泄漏位置,若配合更好的探头定位方法,误差将进一步减小。根据测得的尖峰电流值,能明显分辨9个直径梯度为0.5 mm的圆形泄漏孔,使用测得的9组数据训练神经网络模型,大致得到了尖峰电流值与泄漏孔大小的映射关系,表明根据检测结果能大致估计泄漏孔的大小。

关键词：重力流管道排水管道地电流管道泄漏

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