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| 1 | 我国抗除草剂水稻种质创制研究进展显示文摘水稻是我国最重要的粮食作物之一,稻田杂草是影响水稻产量和品质的主要因素。随着水稻直播稻等轻简化栽培种植面积的日益扩大,各种除草剂的应用日益普遍,除草剂的用量也逐渐增大,稻田抗性杂草及土壤高残留问题也日益严重。因此,加快各类除草剂抗性水稻种质创制与利用具有重要的意义。本文从除草剂的类型及其作用机理、我国抗除草剂水稻种质创制及应用等方面进行了综述,指出了我国抗除草剂水稻种质创制及应用过程中存在的问题,并对今后抗除草剂水稻种质创制及应用的可能发展方向进行了探讨。 | 吴云雨 肖宁 余玲 蔡跃 潘存红 李育红 张小祥 黄年生 周长海 季红娟 戴正元 李爱宏 | 2021 | 植物遗传资源学报2021，22，4： | 14 |
| 2 | 1株抗草甘膦棉花突变体草甘膦抗性的初步鉴定显示文摘目的在于探明1株新的草甘膦抗性棉花突变体是否能用于发掘新的草甘膦抗性基因以及在农业生产中作为抗草甘膦种质的选育和利用的潜在价值。本文采用PCR的方法,在基因组和转录水平上排除了CP4-EPSPS基因对该突变棉株的污染;通过测定莽草酸含量,鉴定了该突变体草甘膦抗性的典型生理特征;通过盆栽试验研究了该突变体苗期对草甘膦抗性的表型。结果显示:不论草甘膦处理与否,突变棉株莽草酸的含量都没有显著的积累,在生理水平上显示出草甘膦抗性植株的显著特点;2叶期时草甘膦处理结果显示,突变棉株的草甘膦抗性表型同孟山都的品种对草甘膦的抗性表型基本一致。在基因组和转录水平的PCR检测结果都排除了突变棉株草甘膦抗性的获得是CP4-EPSPS基因污染造成的。结合对其他已报道的草甘膦抗性基因的类似排除,说明该抗性突变存在着自身特有的分子机理。 | 巩元勇 郭书巧 束红梅 何林池 倪万潮 | 2014 | 棉花学报2014，26，1： | 11 |
| 3 | 抗草甘膦转基因玉米AG16分子特征和抗性鉴定显示文摘抗草甘膦转基因玉米(Zea mays)能有效降低杂草防治成本,具有重要的应用前景。浙江大学转基因抗虫植物和生物安全实验室前期通过农杆菌介导法,以玉米Hi-Ⅱ品种为受体导入新型抗草甘膦基因G10evo,获得不同的抗草甘膦玉米转化系。在此基础上,筛选了具有良好草甘膦抗性的转基因玉米转化系AG16。本研究利用PCR、Western杂交、Southern杂交、ELISA等方法对AG16进行分子特征检测,并对AG16的草甘膦抗性水平进行鉴定。结果表明,G10evo在AG16中为单拷贝插入;G10evo蛋白在AG16的根、茎和叶组织中表达;ELISA分析表明,草甘膦喷洒前,嫩茎中G10evo蛋白的表达量达到9.975μg·g-1。温室中草甘膦抗性测定结果显示,AG16能抗4~8倍田间浓度的草甘膦,远高于草甘膦在实际生产中的使用量。因此,AG16草甘膦抗性水平达到生产需求,具有产业应用潜力,为培育具有自主知识产权的抗草甘膦玉米提供了种质资源。 | 刘苗苗 程家慧 林海燕 沈志成 王志勇 林朝阳 | 2017 | 草业科学2017，34，9： | 6 |
| 4 | CRISPR/Cas9: Development and Application in Rice Breeding显示文摘Rice(Oryza sativa L.)is an important staple food crop worldwide due to its adaptability to different environmental conditions.Because of its great economic and social importance,there is a constant requirement for new varieties with improved agronomic characteristics,such as tolerance to different biotic(such as bacterium,fungus,insect and virus)and abiotic stresses(such as salinity,drought and temperature),higher yield and better organoleptic and nutritional value.Among the new genome editing technologies,the clustered regularly interspaced short palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)(CRISPR/Cas)system allows precise and specific edition in a targeted genome region.It is one of the most frequently used techniques for the study of the function of new genes and for the development of mutant lines with enhanced tolerance to biotic and abiotic stresses,herbicide resistance or improved yield.The wide varieties of applications for this technology include simple non-homologous end joining,homologous recombination,gene replacement,and base editing.In this review,we analyzed how some of these applications have been used in rice cultivars to obtain rice varieties better adapted to current environmental conditions and market requirements. | Fernando Matías ROMERO Andrés GATICA-ARIAS | 2019 | Rice science2019，26，5： | 6 |
| 5 | 禾本科-豆科作物复合种植模式下化学除草的研究现状及展望显示文摘禾本科(Gramineae)-豆科(Leguminosae)作物复合种植模式是世界公认的可持续发展的种植模式,该复合种植模式能够增加作物产量、增强作物抗逆性和改善农田生态环境。玉米(Zea mays)-大豆(Glycine max)间(套)是该复合种植模式的典型代表。《国务院办公厅关于加快转变农业发展方式的意见》([2015] 59)号文件明确指出,在黄淮海地区推广玉米(Zea mays)-花生(Arachis hypogaea)间(套)作和玉米(Zea mays)-大豆(Glycine max)间(套)作,在西南地区推广玉米-大豆间(套)作。近年来,该种植模式发展势头良好,受到农民广泛认可。然而,禾本科、豆科作物复合种植模式包括单子叶、双子叶两种作物。在该复合种植模式下,使用化学除草剂除草面临着一定的困难。本研究围绕禾本科-豆科作物复合种植模式下化学除草剂的使用,介绍了在不同禾本科、豆科作物复合种植模式下,田间杂草的发生情况,简述了化学除草剂的使用现状,探讨了如何能够解决该复合种植模式下杂草危害;展望了今后在禾本科、豆科作物复合种植模式下几种除草方式。通过研究进展的综述为禾本科-豆科作物在复合种植模式下对化学除草剂的使用研究提供依据。 | 金福 陈元凯 周文冠 肖特 任建锐 周颖 刘占鑫 雍太文 杨文钰 | 2019 | 分子植物育种2019，17，17： | 5 |
| 6 | 赤霉素对草甘膦的增效作用及其作用机制显示文摘将系列浓度的赤霉素(GA3)和草甘膦混合喷施玉米幼苗,研究外源GA3对草甘膦喷施效果的影响。结果表明:10~50 mg.L-1GA3与草甘膦混用,比同剂量草甘膦单剂处理玉米幼苗叶色更黄、萎焉更严重,其中50 mg.L-1GA3对草甘膦增效幅度最强。草甘膦和50 mg.L-1GA3混用喷施后,玉米幼苗的株高、地上部鲜质量、叶片叶绿素含量均明显降低,叶片莽草酸、丙二醛(MDA)含量明显增加。 | 李小艳 许晅 李桂俊 夏凯 | 2013 | 南京农业大学学报2013，36，3： | 4 |
| 7 | 抗草甘膦、抗螟虫水稻E1C608的鉴定和重要表型特征分析显示文摘草害和虫害是造成水稻减产的两个重要因素,培育转基因抗除草剂、抗虫水稻是解决这两个问题的有效途径之一。分子鉴定结果表明,以恢复系R608为受体的遗传转化获得了3个独立转基因事件,其中1个为单拷贝转化体,命名为E1C608-3。用ELISA方法检测外源抗草甘膦和抗螟虫蛋白含量的结果表明,E1C608-3的T2代分蘖期的根、茎、叶中EPSPS蛋白含量为120.16-1223.28μg/g,CRY1C蛋白含量为1.23-8.72μg/g,且两个外源蛋白在不同器官中的表达量均表现为叶>茎>根(P<0.01)。T3代测定结果显示,E1C608-3转化体在秧苗期草甘膦的耐受浓度至少为16g/L,其耐受浓度至少是转化受体R608的16倍。T3代抗虫实验结果显示,E1C608-3对稻纵卷叶螟幼虫平均致死率为95.56%,表现出良好的螟虫抗性。T4代农艺性状测定结果显示,E1C608-3与转化受体存在较大差异,但都处在水稻天然种质资源的变异范围之内。以上结果表明,E1C608-3是一个抗除草剂、抗螟虫的水稻新种质。 | 曾强 孟秋成 邓力华 李锦江 于江辉 翁绿水 肖国樱 | 2019 | 中国生物工程杂志2019，39，11： | 3 |
| 8 | Development of herbicide resistance genes and their application in rice显示文摘Rice is one of the most important food crops in the world.Weeds seriously affect the rice yield and grain quality.In recent years,there are tremendous progresses in the research and application of herbicideresistant genes in rice worldwide.This article reviews the working mechanisms of six herbicides(glyphosate,glufosinate,acetolactate synthase inhibitor herbicides,acetyl-Co A carboxylase inhibitor herbicides,hydroxyhenylpyruvate dioxygenase(HPPD)inhibitor herbicides and dinitroaniline herbicides),the resistance mutations of the corresponding herbicide-target genes,and the herbicide detoxification mechanisms by non-target genes.Examples are provided on herbicide-resistant rice materials obtained by transformation of exogenous resistance genes,by artificial mutagenesis and mutant screening,and by modifying the target genes through gene editing.This paper also introduces the current application of herbicide-resistant rice,points out problems that may be caused by utilization of herbicide resistant rice and solutions to the problems,and discusses the future prospects for the development of herbicideresistant rice. | Man Jin Lei Chen Xing Wang Deng Xiaoyan Tang | 2022 | The Crop Journal2022，10，1： | 3 |
| 9 | 肺炎克雷伯氏菌S001草甘膦靶标酶基因的克隆及其抗性验证显示文摘【目的】由于草甘膦在农业生产中的广泛使用,抗草甘膦转基因作物的研究一直是转基因作物研究的热点,其中草甘膦抗性功能新基因的挖掘是核心问题。自然界中微生物种类繁多,基因资源丰富,论文拟从广东地区农田土壤中筛选和鉴定出高抗草甘膦菌株,克隆其草甘膦靶标酶基因并进行抗性水平验证,以期获得高抗草甘膦新基因资源用于抗草甘膦转基因作物的研究。【方法】用含有浓度梯度草甘膦的选择培养基从备选土壤中筛选出具有高抗草甘膦特性的菌株;通过显微观察、革兰氏染色以及16S rDNA序列分析结果对菌株种类进行鉴定;利用RT-PCR技术克隆该菌株的草甘膦靶标酶基因aroAS001,并通过序列比对和系统发育树分析aroAS001序列的基本特征;利用重叠PCR法对aroAS001变异位点进行定点突变获得aroAS001-mut序列后,将aroAS001和aroAS001-mut基因片段转入aroA基因缺陷型菌株DH5α/△aroA中进行基因功能互补和草甘膦抗性水平验证。【结果】分离出一株高抗草甘膦菌株,经形态学和分子生物学鉴定该菌株为肺炎克雷伯氏菌(Klebsiella pneumoniae),命名为kpS001;克隆该菌株草甘膦靶标酶5-烯醇式丙酮酰莽草酸-3-磷酸合成酶(EPSPS)基因aroAS001,序列分析结果显示由该基因所编码的氨基酸序列具有典型的Class I EPSPS特征,但与已报道肺炎克雷伯氏菌的aroA相比其第227位碱基发生突变,致使对应的氨基酸发生变异。对该基因差异位点进行核酸定点突变获得aroAS001-mut基因片段后,将aroAS001和aroAS001-mut基因片段分别转入缺陷型大肠杆菌菌株DH5α/△aroA进行功能互补验证,与对照菌株相比,转入aroAS001和aroAS001-mut的重组菌株均能在含200 mmol·L-1以下浓度草甘膦的培养基中能够正常生长,表现出良好的抗性,之后随着草甘膦浓度增加其生长状态开始受到抑制,当草甘膦浓度达到350 mmol·L-1时,菌株生长基本完全被抑制。【结论】肺炎克雷伯氏菌kpS001菌株是一株高抗草甘膦菌株,由aroAS001编码的草甘膦靶标酶EPSPS属于Class I EPSP合成酶,aroAS001对草甘膦具有优良抗性,能在含350 mmol·L-1以下浓度草甘膦的培养基中生长,该基因可以作为备选基因资源用于抗草甘膦转基因作物的研究。 | 张纯 吴丹丹 冯莉 田兴山 郭爱玲 | 2014 | 中国农业科学2014，47，1： | 3 |
| 10 | 草甘膦为筛选标记的水稻高效遗传转化体系的建立显示文摘【目的】建立以日本晴愈伤组织为受体、草甘膦抗性基因CP4为选择标记基因的高效水稻遗传转化体系。【方法】以粳稻日本晴的成熟胚为试验材料,通过农杆菌介导法,将含有草甘膦抗性基因CP4的P1300载体转化日本晴的愈伤组织,分别用质量浓度为10、20和40 mg/L的草甘膦进行3轮抗性筛选,诱导分化和再生成苗;利用1000 mg/L草甘膦对移栽成活的阳性苗进行抗性鉴定。【结果】获得145株转基因幼苗,PCR检测阳性株为128株,阳性率达88.27%;经过3轮抗性筛选后,转基因幼苗的阳性率提高;移栽成活的99株阳性苗中,65株表现为抗性,抗性率为65.66%。【结论】成功建立了以草甘膦为筛选标记的水稻转基因体系,该体系为水稻育种提供了材料,并对以抗草甘膦基因作为筛选标记的水稻转基因和一些基因功能验证提供了技术支持。 | 袁冰 丁筠 曹含章 吕尊富 李飞飞 | 2021 | 云南农业大学学报(自然科学版)2021，36，4： | 2 |
| 11 | Characteristic analysis of tetra-resistant genetically modified rice显示文摘In this study, the characteristic of three transformants named as B1 C106-1, B1 C106-2, and B1 C106-3 were studied that carried three innate resistant genes Bph14, Bph15, and Xa23, and two enthetic resistant genes Cry1 Ca# and Bar. The five resistant genes were all verified by PCR and the two enthetic genes were identified in single copy insertion by Southern blot. At tillering stage, the Cry1 C and PAT(phosphinothricin acetyl transferase) protein contents in leaf, sheath, and stem of T2 generation were in the similar pattern: leaf>stem>sheath, and showed significant difference(P<0.01) among three organs. The average contents of Cry1 C protein in plant of B1 C106-1, B1 C106-2, and B1 C106-3 were 12.95, 6.57, and 11.30 μg g–1, respectively, and showed significant difference(P<0.01) among them. However, the average contents of PAT in plant of B1 C106-1, B1 C106-2, and B1 C106-3 were 28.54, 27.66, and 28.02 μg g–1, respectively, and there were no significant difference among three transformants. The glufosinate tolerable concentration of three transformants of T3 generation reached at least 6 g L–1, and the mortality of rice leaf rollers were above 97.4% in 5 days after being fed with fresh transformants' leaves. The Cry1 C protein toxicity was also assessed by silkworms, and the mortality of silkworms feeding mulberry leaves smeared with Cry1 C protein extracts of leaves of B1 C106-1, B1 C106-2, and B1 C106-3 were 90, 67.8, and 87.8%, respectively, that were positive correlation(r=0.993) with Cry1 C protein contents in plant of three transformants. The three transformants also maintained high resistance to brown planthopper and bacterial blight as the original version. The above results indicate the tetra-resistant rice germplasm was well-developed by pyramiding innate and enthetic resistant genes in an elite line to provide with resistances of glufosinate, rice leaf roller, brown planthopper, and bacterial blight. | HU Wen-bin DENG Xiang-yang DENG Xiao-xiang DENG Li-hua XIAO You-lun HE Xing-jian FU Xi-qin XIAO Guo-ying | 2018 | Journal of Integrative Agriculture2018，17，3： | 2 |
| 12 | 高度耐受草甘膦菌株JDG-1的分离及其培养条件初探显示文摘通过摇瓶富集培养法从长期喷施草甘膦的土壤中分离出一种草甘膦抗性菌株,命名为JDG-1。该菌株在M9基础盐培养基上最高可以耐受500mmol/L的草甘膦浓度。通过菌株基因组16S rDNA序列及菌落形态学分析,鉴定JDG-1菌株为欧文氏菌属(Erwinia)细菌(GenBank:MH777390)。JDG-1菌株在含100、200mmol/L草甘膦筛选压的LB和M9液体培养基中长势良好。通过pH值梯度和温度梯度培养试验发现,在pH值为7.0和温度为31℃的条件下菌株JDG-1生长最好。抗生素抗药性试验发现JDG-1菌株具有一定水平的氨苄青霉素、链霉素和庆大霉素抗性能力。菌株JDG-1具有较高的草甘膦抗性和嗜药性,可作为草甘膦污染环境的生物修复菌种。 | 肖培英 刘峰 刘义 曹越平 | 2019 | 上海交通大学学报(农业科学版)2019，37，3： | 1 |
| 13 | 利用基因拆分技术培育耐草甘膦转基因水稻的研究显示文摘耐除草剂转基因水稻基因飘流可能产生的环境安全问题是人们关注的焦点之一,并已成为耐除草剂转基因水稻能否在我国生产上发挥效益的限制因素。基因拆分技术能够有效地控制转基因目标性状飘流,为培育耐除草剂转基因水稻提供新的途径和思路。本研究将耐除草剂基因G2-aroA拆分成N端(EPSPSn,1~295aa)和C端(EPSPSc,296~435aa),分别与SspDnaE蛋白内含肽的N端(Intein-N)和C端(Intein-C)连接形成融合基因EPSPSn-In与Ic-EPSPSc,并分别通过农杆菌介导法转入受体材料中花11。Southern杂交证明转基因水稻En-12和Ec-22中外源基因为单拷贝插入。侧翼序列分析证明转基因水稻En-12和Ec-22中外源基因分别插入第2和第6染色体。利用四引物法筛选出转基因水稻En-12和Ec-22的纯合系,并通过有性杂交获得同时含有EPSPSn-In与Ic-EPSPSc的转基因水稻En×Ec。草甘膦抗性分析发现,单独含有1个基因片段的转基因水稻En-12和Ec-22不具有耐受草甘膦特性,而同时含有2个基因片段的转基因水稻En×Ec具有耐受草甘膦的特性,说明拆分后的2个蛋白片段在intein的介导下重新组装成完整有功能蛋白,并赋予转基因水稻耐受草甘膦的特性。转基因水稻En×Ec与含有完整G2-aroA转基因水稻G2-6相比,其耐受草甘膦的能力有所下降,但能够满足生产需求。本研究结果为利用基因拆分技术培育转基因耐草甘膦水稻提供了科学依据,同时也为利用基因工程手段培育转基因杂交稻提供了新的技术平台。 | 董玉凤 王旭静 宋亚亚 靳茜 王志兴 | 2019 | 作物学报2019，45，3： | 1 |
| 14 | Verification of Glyphosate Resistance, Lepidopteran Resistance and Wide Compatibility of Male Sterile Line E1C4008S in Rice显示文摘Weeds and insect pests are two important biotic stresses resulting in yield loss in rice, and wide compatibility is the essential characteristic of breeding inter-subspecific hybrid rice. This study focused on glyphosate resistance, lepidopteran resistance and wide compatibility as well as identification of molecular and some agronomic characteristics of transgenic male sterile line E1 C4008 S. The results indicated that glyphosate resistance gene Epsps^# and lepidopteran resistance gene Cry1 ca^# were transferred into japonica wide compatibility male sterile line 4008 S by Agrobacterium-mediated method, and four independent transformation events named E1 C4008 S-1, E1 C4008 S-2, E1 C4008 S-3 and E1 C4008 S-4 were obtained, in which E1 C4008 S-3 and E1 C4008 S-4 were of single copy insertion. The EPSPS protein contents of E1 C4008 S-3 and E1 C4008 S-4 in different organs were significantly different both in descending order of leaf > stem > root, and ranged from 300.58 to 1410.69 μg/g at the tillering stage. The glyphosate tolerable concentration(dosage) of E1 C4008 S-3 and E1 C4008 S-4 reached at least 16 g/L(54.42 kg/hm^2), and the seeds of E1 C4008 S-4 can germinate normally on the medium containing 1 g/L glyphosate. The CRY1 C protein contents of E1 C4008 S-3 and E1 C4008 S-4 in different organs were significantly different both in descending order of leaf > stem > root, and ranged from 0.62 to 2.43 μg/g at the tillering stage. The larvae mortalities of rice leaf rollers fed on leaves of E1 C4008 S-3 and E1 C4008 S-4 for 5 d were 95.35% and 97.77%, respectively, while the average mortalities of silkworms fed with protein extracts from leaves of E1 C4008 S-3 and E1 C4008 S-4 reached 94.55% and 83.64%, respectively. The results suggested that wide compatibility and evaluated agronomic traits of E1 C4008 S-4 were not significantly changed by insertion of the exogenous genes. Overall, a novel male sterile germplasm with glyphosate resistance, lepidopteran resistance and wide compatibility was verified to be developed in rice. | ZENG Qiang DENG Lihua HU Wenbin HE Xingjian MENG Qiucheng YU Jianghui LI Jinjiang WENG Lushui XIAO Guoying | 2020 | Rice science2020，27，3： | 0 |
| 15 | In maize,co-expression of GAT and GR79-EPSPS provides high glyphosate resistance,along with low glyphosate residues显示文摘Herbicide tolerance has been the dominant trait introduced during the global commercialization of genetically modified(GM)crops.Herbicide-tolerant crops,especially glyphosate-resistant crops,offer great advantages for weed management;however,despite these benefits,glyphosate-resistant maize(Zea mays L.)has not yet been commercially deployed in China.To develop a new bio-breeding resource for glyphosate-resistant maize,we introduced a codon-optimized glyphosate N-acetyltransferase gene,gat,and the enolpyruvyl-shikimate-3-phosphate synthase gene,gr79-epsps,into the maize variety B104.We selected a genetically stable high glyphosate resistance(GR)transgenic event,designated GG2,from the transgenic maize population through screening with high doses of glyphosate.A molecular analysis demonstrated that single copy of gat and gr79-epsps were integrated into the maize genome,and these two genes were stably transcribed and translated.Field trials showed that the transgenic event GG2 could tolerate 9000 g acid equivalent(a.e.)glyphosate per ha with no effect on phenotype or yield.A gas chromatography-mass spectrometry(GC–MS)analysis revealed that,shortly after glyphosate application,the glyphosate(PMG)and aminomethylphosphonic acid(AMPA)residues in GG2 leaves decreased by more than 90%compared to their levels in HGK60 transgenic plants,which only harbored the epsps gene.Additionally,PMG and its metabolic residues(AMPA and N-acetyl-PMG)were not detected in the silage or seeds of GG2,even when far more than the recommended agricultural dose of glyphosate was applied.The co-expression of gat and gr79-epsps,therefore,confers GG2 with high GR and a low risk of herbicide residue accumulation,making this germplasm a valuable GR event in herbicide-tolerant maize breeding. | Shengyan Li Pengcheng Li Xiangyin Li Ning Wen Yinxiao Wang Wei Lu Min Lin Zhihong Lang | 2023 | aBIOTECH2023，4，4： | 0 |
| 16 | 牛筋草EPSPS基因启动子对草甘膦的响应显示文摘杂草对草甘膦抗药性机理及抗草甘膦转基因作物研究主要围绕草甘膦靶标酶EPSPS展开,EPSPS基因表达量增加是杂草抵抗草甘膦胁迫的主要分子机制之一,但EPSPS基因表达调控机理及其对草甘膦的响应却鲜有研究。本研究根据Gen Bank中牛筋草EPSPS基因启动子序列设计引物,从牛筋草基因组DNA中扩增EPSPS基因启动子序列及其5'端缺失序列;将各启动子片段构建到含绿色荧光蛋白基因GFP的p35S-GFP载体上,在农杆菌的介导下,利用洋葱表皮细胞瞬时转染体系分析EPSPS基因启动子活性及对草甘膦的响应,结果表明,各启动子片段均能启动下游基因的表达,具有较强的启动子活性,可作为较好的实验材料用于抗草甘膦转基因作物的研究。此外,EPSPS基因启动子能够响应草甘膦信号,调节下游GFP基因的表达,响应元件可能存在于3'端的345 bp内。研究结果有助于进一步丰实杂草对草甘膦抗药性机理及抗草甘膦转基因作物研究的理论基础。 | 张纯 王田 冯莉 田兴山 | 2017 | 分子植物育种2017，15，10： | 0 |
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