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Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

《农业科学与工程前沿（英文）》 2019年 第6卷 第1期   页码 66-72 doi: 10.15302/J-FASE-2018229

摘要：

Porcine viral diarrhea is an acute and highly contagious enteric disease of pigs that causes huge economic losses worldwide. Porcine epidemic diarrhea virus (PEDV) and transmissible gastroenteritis virus (TGEV) are the main pathogens responsible for piglet viral diarrhea. However, currently there is no specific drug available for the effective treatment of viral diarrhea. Therefore, it is necessary to seek an effective method to diminish PEDV and TGEV infection rates. RNA interference has been applied successfully to inhibit the virus replication. It provides a potential strategy for breeding resistant pigs. In this study, four promoters and four short hairpin RNA (shRNA) vectors with LoxP sites at each end of the selectable marker genes were constructed to target PEDV and TGEV. These vectors were then transfected into porcine fetal fibroblasts, G418 resistant transfectants were confirmed by PCR and transgenic SCNT porcine blastocysts were obtained. These results have paved the way for future production of marker-free transgenic resistant to PEDV and TEGV pigs by SCNT.

关键词： piglet diarrhea     RNAi     PEDV     TGEV     transgenic resistance breeding    

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油菜转基因育种研究进展

官春云

《中国工程科学》 2002年 第4卷 第8期   页码 34-39

摘要：

简要介绍了国外油菜转基因育种情况，包括转除草剂抗性基因，病原菌抗性基因，耐重金属基因， 影响种子贮藏产物基因，影响繁殖特性基因，医药和工业产品基因，以及启动子调节基因等。较详细介绍了湖 南农业大学在Bt毒蛋白基因转化甘蓝型油菜育成抗虫新品系、基因工程雄性不育系、恢复系选育和杂种优势利 用，以及反义FAD2基因转化甘蓝型油菜双低品种提高油酸含量的研究结果；浙江省农科院利用反义PEP基因 转化甘蓝型油菜提高种子含油量的研究结果。

关键词： 油菜     转基因     育种    

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Breeding for the resistance to

Hongxiang MA, Xu ZHANG, Jinbao YAO, Shunhe CHENG

《农业科学与工程前沿（英文）》 2019年 第6卷 第3期   页码 251-264 doi: 10.15302/J-FASE-2019262

摘要：

With the changes of climate and cultivation systems, the head blight (FHB) epidemic area in China has extended since 2000 from the reaches of the Yangtze River to the north and west winter wheat region. Breeding for FHB resistance in wheat is an effective way to control the disease. Chinese wheat breeders commenced research on FHB in the 1950s. Sumai 3, Ning 7840, Yangmai 158, Ningmai 9 and other cultivars with improved FHB resistance were developed through standard breeding methods and widely applied in production or breeding programs. In addition to intervarietal crosses, alien germplasm was used to improve FHB resistance of wheat. Addition, substitution and translocation lines with alien chromosomes or chromosome fragments were created to enhance FHB resistance. Somaclonal variation was also used to develop a FHB resistant cv. Shengxuan 3 and other cultivars with moderate resistance to FHB were released by such methods. QTL (quantitative trait loci) for FHB resistance were characterized in cultivars originating from China. The major QTL, , was identified on chromosome 3BS in Sumai 3, Ning 894037, Wangshuibai and other Chinese resistant sources. Diagnostic molecular markers for 1 have been applied in wheat breeding and breeding lines with improved FHB resistance and desirable agronomic traits have been obtained. However, breeding for FHB resistance is a long-term task, new technologies are likely to increase the efficiency of this process and better FHB resistance of new cultivars is expected to be achieved within the next decade.

关键词： breeding     Fusarium head blight     wheat    

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Transgenic technologies in cassava for nutritional improvement and viral disease resistance: a key strategy

Maliwan NACONSIE,Peng ZHANG

《农业科学与工程前沿（英文）》 2016年 第3卷 第4期   页码 285-294 doi: 10.15302/J-FASE-2016119

摘要： As a major staple food source in Africa and other tropical developing countries, cassava ( ) provides basic sustenance for many subsistence farmers. However, cassava roots mainly accumulate starch with limited contribution of other nutrients such as proteins and vitamins. Also, two viral diseases, cassava mosaic disease (CMD) and cassava brown streak disease (CBSD), cause great losses in cassava production in sub-Saharan Africa and the Indian sub-continent. Genetic engineering provides promising approaches to improve nutritional value and increase resistance to viral diseases in cassava. This report presents several successful case studies on engineering protein content by overexpression of nutritious storage proteins and improving cassava resistance to viral diseases by RNA interference. Perspectives on the sustainable acquisition of new knowledge and development of biotechnology to solve these bottlenecks are discussed.

关键词： Manihot esculenta     protein enhancement     RNA interference     transgenesis     virus resistance    

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Progress in breeding for resistance to Ug99 and other races of the stem rust fungus in CIMMYT wheat germplasm

Sridhar BHAVANI, David P. HODSON, Julio HUERTA-ESPINO, Mandeep S. RANDHAWA, Ravi P. SINGH

《农业科学与工程前沿（英文）》 2019年 第6卷 第3期   页码 210-224 doi: 10.15302/J-FASE-2019268

摘要：

Races belonging to the Ug99 (TTKSK) lineage of the wheat stem rust fungus, carrying complex virulence combinations, and their migration to countries in Africa, Middle East and Asia continue to pose a significant threat to global wheat production. The rapid spread of additional races, e.g., TKTTF or the Digalu lineage, in several countries causing localized epidemics reminds us of the vulnerability of wheat germplasm to stem rust disease, a formidable foe referenced as early as biblical times. A global rust monitoring system reflecting increased surveillance efforts has identified 13 races within the Ug99 lineage in 13 countries and unrelated lineages are emerging, spreading and posing serious threats to wheat production. Race TKTTF has caused localized epidemics in Ethiopia and its variants have been recently implicated in stem rust outbreaks in Europe. Concerted research efforts have resulted in the identification of several new resistance (APR) genes and gene combinations for use in breeding. Combining multiple adult plant resistance genes in high-yielding backgrounds and discovery of new quantitative trait loci conferring stem rust resistance has progressed in the recent years, enhancing the durability of resistance. Effective gene stewardship and new generation breeding materials and cultivars that combine multiple race-specific or minor to intermediate effect APR genes, complemented by active surveillance and monitoring, have helped to limit major epidemics and increase grain yield potential in key target environments.

关键词： adult plant resistance     black rust     race-specific resistance     Triticum aestivum    

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大豆胞囊线虫抗性研究现状及其对大豆育种的启示 Review

Guiping Yan,Richard Baidoo

《工程（英文）》 2018年 第4卷 第4期   页码 534-541 doi: 10.1016/j.eng.2018.07.009

摘要：

大豆胞囊线虫（SCN，Heterodera glycines）是危害全球大豆生产最严重的线虫。作物轮作和培育抗线虫品种是防治线虫的主要方法。随着越来越多能够克服常用抗源的毒性SCN 群体出现，有必要寻找更多新的抗源或拓宽抗性遗传背景。本文综述了大豆抗SCN 基因的研究进展，以及这些基因如何相互作用，从而使大豆能够抗SCN。本文还提供了最新分子作图和分子标记，可用于不同抗性品系和品种的大规模选择和鉴定，以加速常规育种计划。深入了解SCN 寄生蛋白和大豆对病原体的抗性是通过基因修饰、基因叠加、回交或基因工程引入新的抗源而使抗源多样化的关键。

关键词： 大豆胞囊线虫     Heterodera glycines     抗性     分子育种    

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regions included in a bicistronic vector enhances and stabilizes follistatin gene expressions in both transgeniccells and transgenic mice

Xiaoming HU,Jing GUO,Chunling BAI,Zhuying WEI,Li GAO,Tingmao HU,Shorgan BOU,Guangpeng LI

《农业科学与工程前沿（英文）》 2016年 第3卷 第1期   页码 87-96 doi: 10.15302/J-FASE-2016087

摘要： In the present study, follistatin ( ) gene expression vectors with either a bicistronic gene transfer cassette alone, or a bicistron gene cassette carrying a matrix attachment region (MAR) were constructed and transfected to bovine fetal fibroblasts. Evaluations of both the integration and expression of exogenous indicated that the pMAR-CAG-FST-IRES-AcGFP1-polyA-MAR (pMAR-FST) vector had higher capacity to form monoclonal transgenic cells than the vector without MAR, though transient transfection and integration efficiency were similar with either construct. Remarkably, protein expression in transgenic cells with the pMAR-FST vector was significantly higher than that from the bicistronic vector. Exogenous was expressed in all of the pMAR-FST transgenic mice at F , F and F . Total muscle growth in F mice was significantly greater than in wild-type mice, with larger muscles in fore and hind limbs of transgenic mice. pMAR-FST transgenic mice were also found with more evenly distributed muscle bundles and thinner spaces between sarcolemma, which suggests a correlation between transgene expression-associated muscle development and the trend of muscle growth. In conclusion, a pMAR-FST vector, which excluded the resistant genes and frame structure, enhances and stabilizes gene expressions in both transfected cells and transgenic mice.

关键词： safety of transgenic     bicistron gene transfer body     transgenic mice     muscle development    

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美国转基因大西洋鲑产业化对我国的启示

胡炜,朱作言

《中国工程科学》 2016年 第18卷 第3期   页码 105-109 doi: 10.15302/J-SSCAE-2016.03.018

摘要：

2015年11月19日，美国食品药品监督管理局（FDA）批准了转全鱼生长激素基因（gh）大西洋鲑为第一种可供食用的转基因动物产品，从而取得世界转基因动物育种研究与产业化的历史性突破。经过了长达20年的严格审核，转gh大西洋鲑才获准产业化，不仅充分说明了对包括转基因鱼在内的创新产业的管理决策尤其需要尊重科学规律和客观事实；而且表明基础研究只是产业创新的因素之一，转基因知识的科学普及与企业的积极参与是推动高新技术发展和创新产业形成不可或缺的重要条件；高屋建瓴地制定我国基因改良农业品种市场准入的条例和操作细则已经刻不容缓。

关键词： 转基因     大西洋鲑     产业化     启示    

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Breeding for pre-harvest sprouting resistance in bread wheat under rainfed conditions

Muhammad ZEESHAN, Waheed ARSHAD, Muhammad Imran KHAN, Shiraz ALI, Ali NAWAZ, Amina BATOOL, Muhammad TARIQ, Muhammad Imran AKRAM, Muhammad Amjad ALI

《农业科学与工程前沿（英文）》 2018年 第5卷 第2期   页码 253-261 doi: 10.15302/J-FASE-2018218

摘要： Pre-harvest sprouting in wheat is the germination of seeds within the spikes when rains occur after or during grain ripening, which occurs commonly in the barani tract of Pakistan. Therefore, 10 cultivars and five advanced lines of spring bread wheat were evaluated for pre-harvest sprouting resistance. After natural rainfall, seeds were immediately collected from the wet spikes and tested for germinating ability. Three different germination tests were applied to hand-threshed seed: (1) spikes threshed on the day of sampling and germination tested immediately, (2) spikes threshed on the day of sampling and germination tested 1 week later, and (3) spikes threshed 1 week after sampling and germination test immediately after threshing. Seeds and spikes kept for 1 week were place on blotting paper at room temperature. Cultivars BARS-09, 09FJ17, Doukkala-12, NARC-09 and Ouassou-20 exhibited higher sprouting resistance while other genotypes were susceptible to pre-harvest sprouting in each of the three tests. A diallel crossing was conducted with six susceptible and two resistant genotypes to assess the genetic behavior of pre-harvest sprouting resistance. The combining ability (CA) demonstrated a higher proportion of additive genetic effects for sprouting resistance, because of higher variance of general and specific CA for both parameters under study. Doukkala-12 and BARS-09 showed increased pre-harvest sprouting resistance in their F descendants.

关键词： bread wheat     germination index     general combining ability     pre-harvest sprouting     specific combining ability    

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Comments on this special issue on “Wheat Genetics and Breeding”

Zhonghu HE, Xu LLU

《农业科学与工程前沿（英文）》 2019年 第6卷 第3期   页码 309-309 doi: 10.15302/J-FASE-2019264

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Highlights of special issue on “Wheat Genetics and Breeding”

Zhonghu HE, Zhendong ZHAO, Shunhe CHENG

《农业科学与工程前沿（英文）》 2019年 第6卷 第3期   页码 207-209 doi: 10.15302/J-FASE-2019263

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Genomics and genetic breeding in aquatic animals: progress and prospects

Wenteng XU, Songlin CHEN

《农业科学与工程前沿（英文）》 2017年 第4卷 第3期   页码 305-318 doi: 10.15302/J-FASE-2017154

摘要： Genomics focuses on dissection of genome structure and function to provide a molecular basis for understanding the genetic background. In a pivotal step, the expense of whole genome sequencing has been largely eliminated by the rapid updating of sequencing technology, leading to increasing numbers of decoded genomes of aquatic organisms, driving the aquaculture industry into the genomic era. Multiple aquatic areas have been influenced by these findings, such as accelerated generation shift in the seed industry and the process of breeding improved lines. In this article, we have summarized the latest domestic and international progress of aquatic animals in nine aspects, including WGS and fine mapping, construction of high density genetic/physical maps, trait-related marker/genes screening, as well as sex control, genome editing, and other molecular breeding technologies. Finally, the existing problems in this field have been discussed and five future counter measures have been proposed accordingly.

关键词： aquaculture     genomics     genetic breeding     progress    

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Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Dan LU,Shengzhe SHANG,Shen LIU,Ying WU,Fangfang WU,Tan TAN,Qiuyan LI,Yunping DAI,Xiaoxiang HU,Yaofeng ZHAO,Ning LI

《农业科学与工程前沿（英文）》 2014年 第1卷 第3期   页码 179-184 doi: 10.15302/J-FASE-2014020

摘要： Butyrylcholinesterase (BCHE) is a natural bioscavenger that protects humans against organophosphate toxicity. Due to the limited yield of human BCHE (hBCHE) when purifying from human plasma, it is necessary to find an alternative method to produce this protein. One potential method is to produce transgenic livestock that make modified milk containing high concentration of hBCHE. In this study, we cloned the gene into a human lactoferrin (hLF) bacterial artificial chromosome (BAC) construct to make a hLF-hBCHE BAC construct. Subsequently, we injected the BAC construct into pronuclei of mouse fertilized embryos and generated transgenic mice. Expression analysis showed that recombinant hBCHE (rhBCHE) was expressed efficiently in the mammary gland of the transgenic mice and the concentration of rhBCHE in the milk of individual mice ranged from 76±12 to 159±28 mg·L . Protein function tests showed that rhBCHE has the same enzymatic activity as the native hBCHE. Our results pave the way for making transgenic livestock to produce large quantities of rhBCHE.

关键词： recombinant human butyrylcholinesterase (rhBCHE)     human lactoferrin bacterial artificial chromosome (hLF BAC)     transgenic mice     milk    

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Wheat research and breeding in the new era of a high-quality reference genome

Rudi APPELS

《农业科学与工程前沿（英文）》 2019年 第6卷 第3期   页码 225-232 doi: 10.15302/J-FASE-2019265

摘要：

The publications of the International Wheat Genome Sequencing Consortium (IWGSC) released in August 2018 are reviewed and placed into the context of developments arising from the availability of the high-quality wheat genome assembly.

关键词： assembly technology     molecular markers     polyploidy     transcript networks     wheat genome    

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Genetic study and molecular breeding for high phosphorus use efficiency in maize

null

《农业科学与工程前沿（英文）》 2019年 第6卷 第4期   页码 366-379 doi: 10.15302/J-FASE-2019278

摘要：

Phosphorus is the second most important macronutrient after nitrogen and it has many vital functions in the life of plants. Most soils have a low available P content, which has become a key limiting factor for increasing crop production. Also, low P use efficiency (PUE) of crops in conjunction with excessive application of P fertilizers has resulted in serious environmental problems. Thus, dissecting the genetic architecture of crop PUE, mining related quantitative trait loci (QTL) and using molecular breeding methods to improve high PUE germplasm are of great significance and serve as an efficient approach for the development of sustainable agriculture. In this review, molecular and phenotypic characteristics of maize inbred lines with high PUE, related QTL and genes as well as low-P responses are summarized. Based on this, a breeding strategy applying genomic selection as the core, and integrating the existing genetic information and molecular breeding techniques is proposed for breeding high PUE maize inbred lines and hybrids.

关键词： maize     phosphorus use efficiency     quantitative trait loci     genetic study     molecular breeding     genomic selection    

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标题 作者 时间 类型 操作

Construction of multiple shRNA vectors targeting PEDV and TGEV and production of transgenic SCNT porcine

Jianwen CHEN, Kaiyuan PAN, Zhen CHEN, Biao DING, Dandan SONG, Wenbin BAO, Yunhai ZHANG

期刊论文

油菜转基因育种研究进展

官春云

期刊论文

Breeding for the resistance to

Hongxiang MA, Xu ZHANG, Jinbao YAO, Shunhe CHENG

期刊论文

Transgenic technologies in cassava for nutritional improvement and viral disease resistance: a key strategy

Maliwan NACONSIE,Peng ZHANG

期刊论文

Progress in breeding for resistance to Ug99 and other races of the stem rust fungus in CIMMYT wheat germplasm

Sridhar BHAVANI, David P. HODSON, Julio HUERTA-ESPINO, Mandeep S. RANDHAWA, Ravi P. SINGH

期刊论文

大豆胞囊线虫抗性研究现状及其对大豆育种的启示

Guiping Yan,Richard Baidoo

期刊论文

regions included in a bicistronic vector enhances and stabilizes follistatin gene expressions in both transgeniccells and transgenic mice

Xiaoming HU,Jing GUO,Chunling BAI,Zhuying WEI,Li GAO,Tingmao HU,Shorgan BOU,Guangpeng LI

期刊论文

美国转基因大西洋鲑产业化对我国的启示

胡炜,朱作言

期刊论文

Breeding for pre-harvest sprouting resistance in bread wheat under rainfed conditions

Muhammad ZEESHAN, Waheed ARSHAD, Muhammad Imran KHAN, Shiraz ALI, Ali NAWAZ, Amina BATOOL, Muhammad TARIQ, Muhammad Imran AKRAM, Muhammad Amjad ALI

期刊论文

Comments on this special issue on “Wheat Genetics and Breeding”

Zhonghu HE, Xu LLU

期刊论文

Highlights of special issue on “Wheat Genetics and Breeding”

Zhonghu HE, Zhendong ZHAO, Shunhe CHENG

期刊论文

Genomics and genetic breeding in aquatic animals: progress and prospects

Wenteng XU, Songlin CHEN

期刊论文

Expression of recombinant human butyrylcholinesterase in the milk of transgenic mice

Dan LU,Shengzhe SHANG,Shen LIU,Ying WU,Fangfang WU,Tan TAN,Qiuyan LI,Yunping DAI,Xiaoxiang HU,Yaofeng ZHAO,Ning LI

期刊论文

Wheat research and breeding in the new era of a high-quality reference genome

Rudi APPELS

期刊论文

Genetic study and molecular breeding for high phosphorus use efficiency in maize

null

期刊论文