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转基因水稻基因飘流研究十年回顾
引用本文:贾士荣,袁潜华,王丰,姚克敏,裴新梧,胡凝,王志兴,王旭静,柳武革,钱前.转基因水稻基因飘流研究十年回顾[J].中国农业科学,2014,47(1):1-10.
作者姓名:贾士荣  袁潜华  王丰  姚克敏  裴新梧  胡凝  王志兴  王旭静  柳武革  钱前
作者单位:1.中国农业科学院生物技术研究所,北京 100081;
2.海南大学农学院/海南省热带生物资源可持续利用重点实验室,海口 570228;
3.广东省农业科学院水稻研究所,广州 510640;
4.南京信息工程大学应用气象学院,南京210044;
5.中国水稻研究所,杭州310006
基金项目:国家“973”计划项目(2001CB109003,2007CB109202)、国家“863”计划项目(2002AA2121493)、国家转基因新品种培育重大专项(2009ZX08011-010B,2009ZX08012-019B,2013ZX08011-001)
摘    要:中国是世界最大的水稻生产国和亚洲栽培稻的起源中心之一。随着中国转基因水稻研发的快速发展,需要研究水稻转基因飘流可能对环境和食品安全带来的潜在风险。基因飘流及其数据是对转基因水稻进行科学评估和监管的重要参数。为此,从2002年开始组建了研究团队,对转基因水稻的基因飘流进行了为期10年的系统研究。取得的结果主要包括:(1)阐明了水稻基因飘流的基本规律,揭示了影响水稻基因飘流的生物学和气象学主控因子。沿水稻开花期的主流风向,采用长方形田间试验设计,分别在三亚、广州和杭州3个点2-3个生长季,研究了纯合转bar基因花粉供体L201或B2(姐妹系,抗除草剂Basta)向19个非转基因受体(包括不育系、常规稻品种、杂交稻F1和普通野生稻)在不同距离上的基因飘流率,明确了转基因向不育系的飘流率最高,向普通栽培稻品种的基因飘流率最低(相邻种植时小于1%或0.1%),向普通野生稻的基因飘流率介于不育系和常规稻之间,向不育系的最大基因飘流率比向普通野生稻和栽培稻要大1-3个数量级;基因飘流率随距离增加呈负指数曲线衰减,且存在急剧降低的"拐点","拐点"的距离与试验点水稻开花期的风速密切相关,广州和杭州为1-2 m,三亚约为5 m;采用圆形、以花粉供体为中心的田间试验设计,以异交结实率很高的不育系博A作受体,清晰地解析了风向与基因飘流率的数值关系,主流和次主流风向下游4个扇区的基因飘流事件累计达90%-96%,而逆风向和侧逆风向4个扇区仅为4%-10%。综上所述,水稻转基因飘流率与常规育成品种间的异交率(一般在1%以下)基本相同,在数量级上转基因并未增加新的风险。(2)建立了以气象资料为参数的水稻花粉扩散和基因飘流普适模型,计算和预测了中国南方稻区17省、市的最大基因飘流阈值距离(maximum threshold distances,MTDs)。受东南季风和地形地貌的影响,中国南方稻区MTDs的空间分布特征为:东西之间有自东向西逐渐减小的趋势,南北之间首先在南方丘陵地区逐渐减小、越过南岭后再向东南沿海地区逐渐增大。(3)利用人工构建的普通野生稻与基因(Bt或bar)飘流后代栽/野F1杂种混栽群体,经多年多代跟踪观察,分析了转基因飘流至普通野生稻后的命运,发现栽/野F1杂种在3-5年后完全消失,混栽群体中检测不到外源的Bt和bar,有理由推测普通野生稻具有自我保护的机制。(4)研究了小规模田间试验中采用花期隔离和布帐隔离措施降低水稻基因飘流率的效果;调查了海南、广东、广西普通野生稻原生境居群与相邻种植的栽培稻花期相遇情况,建立了相应的数据库;研究了基因拆分技术作为生物学限控措施从根本上限控基因飘流的效果;以本研究的结果及对国际上主要农作物基因飘流的调研数据为基础,提出了在水稻基因飘流风险评估和监管中采用分类管理和阈值管理的原则。在10年回顾和科学分析的基础上,对未来研究的重点也进行了展望。

关 键 词:水稻    普通野生稻    转基因    花粉扩散    基因飘流    风险评估    风险管理
收稿时间:2013-07-27

What We Have Learnt in Ten Years′ Study of Rice Transgene Flow
JIA Shi-Rong-,YUAN Qian-Hua-,WANG Feng-,YAO Ke-Min-,PEI Xin-Wu-,HU Ning-,WANG Zhi-Xing-,WANG Xu-Jing-,LIU Wu-Ge-,QIAN Qian-.What We Have Learnt in Ten Years′ Study of Rice Transgene Flow[J].Scientia Agricultura Sinica,2014,47(1):1-10.
Authors:JIA Shi-Rong-  YUAN Qian-Hua-  WANG Feng-  YAO Ke-Min-  PEI Xin-Wu-  HU Ning-  WANG Zhi-Xing-  WANG Xu-Jing-  LIU Wu-Ge-  QIAN Qian-
Affiliation:1.Biotechnology Research Institute, Chinese Academy of Agricultural Sciences, Beijing 100081; 2.College of Agriculture, Hainan University/Hainan Key Laboratory for Sustainable Utilization of Tropical Bioresources, Haikou 570228; 3.Rice Research Institute, Guangdong Academy of Agricultural Sciences, Guangzhou 510640; 4.College of Applied Meteorology, Nanjing University of Information Science & Technology, Nanjing 210044; 5.China National Rice Research Institute, Hangzhou 310006
Abstract:China is the largest rice producer worldwide and is one of the origins of Asian cultivated rice as well. Along with the rapid development of transgenic rice in China, the potential impact of rice transgene flow on the environment and food safety has become one of the major concerns. Gene flow is an important parameter in the risk assessment and regulation of transgenic rice on the scientific basis. In accordance with this situation, we have formed a team and systematically studied the rice transgene flow since 2002. The results obtained in recent ten years are as following: (1) the patterns of transgene flow and the major biological and meteorological factors controlling rice gene flow have been elucidated. Following the prevailing wind direction in rice flowering period, a rectangular design of field experiments were conducted at 3 locations(Sanya, Hainan Island; Guangzhou, Guangdong; and Hangzhou, Zhejiang)in 2-3 years by using a homozygous transgenic line L201 or B2 (sister lines) with bar gene inserted, resistant to herbicide Basta, as a pollen donor, and totally 19 non-transgenic rice as recipients, including male sterile (ms) lines, common rice cultivars (CRC), F1 hybrid rice, and common wild rice (Oryza rufipogon). Results indicated that the frequency of transgene flow to ms lines was the highest, while gene flow to CRC and F1 hybrids was the lowest (less than 1% or 0.1% at parallel plantation). The frequency of transgene flow to O. rufipogon was in between. By comparison, the maximum frequency of gene flow to ms lines is one to three orders of magnitude higher than that to O. rufipogon and CRC. Gene flow frequency decreased exponentially as the distance increase, with a sharp cut-off point at about 1-2 m in Guangzhou and Hangzhou, while it was approximately 5m in Sanya. It indicates that the sharp cut-off point is closely related to the wind speed during rice flowering period at a given location. By using a concentric circle design of field experiment and an ms line BoA with higher outcrossing rate as a recipient, we have been able to clearly quantify the relationship between the gene flow frequency and the wind direction. On the basis of the cumulative data in eight compass sectors, 90%–96% of the cumulative gene flow events occurred in the four downstream prevailing wind sectors, while it was only 4%–10% in the four lateral and upstream prevailing wind sectors. In short, a general conclusion is that the order of magnitude of transgene flow frequency is basically the same as the outcrossing rate of the CRC (generally less than 1%), which means the gene transfer has not added a new additional risk. (2) By using historical meteorological data as an input, a regional applicable model of rice pollen dispersal and gene flow has been established, which is successively used to predict the maximum threshold distances (MTDs) of gene flow in 17 provinces of southern rice growing area in China. The feature of spatial distribution of MTDs shows: from east to west, MTDs gradually decrease; from north to south, MTDs first decrease in the hilly region and then increase again along the southeast costal region. Reason for it is that the spatial distribution is dramatically influenced by the southeast monsoon (seasonal wind direction from the southeast) and the landform structure. (3) We have artificially constructed two mixed populations of O. rufipogon with F1 hybrids of CRC/O. rufipogon derived from transgene (either Bt or bar) flow to investigate the long-term fate of the transgene integrated into common wild rice. It was found that the F1 hybrids of CRC/O. rufipogon totally disappeared within 3-5 years and the Bt or bar gene was not detectable in the mixed population. It is reasonable to speculate that the common wild rice possesses a mechanism of self-protection. (4) The effectiveness in reducing transgene flow by using flowering isolation or a protective cloth-screen in small-scale field trials of transgenic rice was studied and the results discussed. To investigate the degree of flowering synchronization of CRC and O. rufipogon populations in adjacent plantation, a survey has been conducted in Hainan, Guangdong and Guangxi provinces which allow us to establish a corresponding database. In order to further eliminate the rice transgene flow, a biological containment measure - gene split approach has been established. Based on the data obtained in this study and a survey on the internationally published data of gene flow from major crops, we have proposed to use principles of classification management and threshold-value management in the risk assessment and regulation of transgenic rice. Meanwhile, the progress and prospective of rice gene flow are also discussed in this paper.
Keywords:rice (Oryza sativa L    common wild rice (O  rufipogon  transgene  pollen dispersal  gene flow  risk assessment  risk management
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