CN108552214A - 一种抗草甘膦玉米用草甘膦包衣剂及其应用 - Google Patents
一种抗草甘膦玉米用草甘膦包衣剂及其应用 Download PDFInfo
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Abstract
本申请属于玉米栽培技术领域,具体涉及一种用于抗草甘膦转基因玉米用草甘膦包衣剂及其应用。所述包衣剂为含有草甘膦的种衣剂,所述种衣剂为克.醇.福美双、克.戊.福美双或噻虫嗪,所述包衣剂中草甘膦质量浓度为0.5~4%。所述抗草甘膦玉米用草甘膦包衣剂在玉米栽培中的应用,用于具有草甘膦抗性的玉米品种。本申请所提供草甘膦在包衣剂中的新应用方式,不仅能用于检测目标品种的抗草甘膦基因的功能效率、鉴定抗草甘膦作物亲本纯度和杂交种制种纯度,而且其可以与实际生产紧密结合,可以在玉米生产中实现杂交种田间自动去除杂株,从而为抗草甘膦玉米品种的推广、单粒播种和优质高产奠定基础,因而具有较好地实用价值和推广应用意义。
Description
技术领域
本申请属于玉米栽培技术领域,具体涉及一种用于抗草甘膦转基因玉米用草甘膦包衣剂及其应用。
背景技术
玉米是中国第一大粮食作物,常年播种面积在3500万公顷左右。2016年,全球转基因作物种植面积达到1.851亿公顷,其中耐除草剂转基因作物种植8 650万公顷,占全球转基因作物总种植面积的47%。由于我国有较为严格的转基因作物安全评价管理程序,目前除了进口用作加工原料外,尚未批准任何转基因抗除草剂玉米商业化种植。至今,具有我国自主知识产权的最有望商业化推广的抗除草剂玉米转化事件为浙江大学的双抗12-5(抗虫、抗草甘膦除草剂)和中国农业大学的CC-2(抗草甘膦除草剂),正在申请安全证书。
由于转基因品种具有巨大的商业价值,因此采用合适方法对转基因作物品种进行鉴定是其推广应用的重要前提。在这方面,Main等报道了通过草甘膦浸种法鉴定抗草甘膦大豆(Main C.L.et.al., A novel approach to determine the glyphosate toleranttrait in soybeans, Journal of agricultural and food chemistry, 2004, 52(5):1224-1227),燕树锋报道了通过草甘膦浸种法鉴定抗草甘膦棉花和玉米(燕树锋等, 通过草甘膦浸种鉴定抗草甘膦棉花, 分子植物育种, 2016, (3): 604-607;一种抗草甘膦玉米品种鉴定方法,CN106576829A)。
但实际应用中发现,转基因玉米品种在生产应用时使用草甘膦浸种法进行筛选鉴定存在一些不易克服的技术缺陷,主要是:(1)由于适应机械化操作的需要以及劳动力成本的上升等原因,玉米生产上大多不再进行浸种处理,因此使得草甘膦浸种法无法在实际生产中推广应用;(2)现有玉米生产上大多采用的是单粒播种的栽培方式,针对这种栽培方式,如何能够在抗草甘膦转基因玉米播种早期即可去除制种中杂株,并根据制种纯度及时补足每亩株数,从而保障玉米的高产优质,这也需要对现有鉴定方法进行改进和发展。
发明内容
本申请主要目的是提供草甘膦在包衣剂中的应用方式,从而为抗草甘膦玉米品种适宜机械化和规模化栽培奠定基础。
本申请所采取的技术方案详述如下。
一种抗草甘膦玉米用草甘膦包衣剂,所述包衣剂为含有草甘膦的种衣剂,所述种衣剂具体例如为商品化的克.醇.福美双(克百威+三唑醇+福美双)、克.戊.福美双(克百威+戊唑醇+福美双)或噻虫嗪,所述包衣剂中草甘膦质量浓度为0.5~4%,较优使用浓度为0.5~2.0%,进一步优选使用浓度为1.0~2.0%;使用该包衣剂包裹处理玉米种子后,具有草甘膦抗性的玉米品种的发芽、成苗和正常生长等不受影响,而不具有草甘膦抗性的玉米品种则无法发芽、或者无法正常成苗,从而可以及时在田间自动剔除制种不纯的杂株,并可根据室内鉴定的杂交种纯度在第一次播种时即可补足种子量,不用再次补种,同时便于玉米种植后期的统一管理等。
所述抗草甘膦玉米用草甘膦包衣剂在玉米栽培中的应用,用于具有草甘膦抗性的玉米品种,具体例如为抗草甘膦玉米双抗12-5,使用后即可在田间有效去除制种杂株,仅保留具草甘膦抗性的玉米杂交种,从而便于集中和统一管理;具体应用时,克.醇.福美双(药剂和种子按照质量比1:50的比例直接进行包衣)、克.戊.福美双(将粉剂按照质量比1:4兑水混匀后,再将药剂和种子按照质量比1:50的比例进行包衣)和噻虫嗪(按照体积比1:1兑水混匀后,再将药剂和种子按照质量比1:50的比例进行包衣)。
总体而言,本申请所提供草甘膦在包衣剂中的新应用方式,不仅能用于检测目标品种的抗草甘膦基因的功能效率、鉴定抗草甘膦作物亲本纯度和杂交种制种纯度,而且其可以与实际生产紧密结合,可以在玉米生产中实现杂交种田间自动去除杂株,从而为抗草甘膦玉米品种的推广、单粒播种和优质高产等奠定基础,因而具有较好地实用价值和推广应用意义。
附图说明
图1为含草甘膦的种衣剂包衣对双抗12-5、联创505和郑单958发芽率的影响(7d),注:种衣剂为克.醇.福美双;
图2为含草甘膦的种衣剂包衣对双抗12-5、联创505和郑单958正常苗率的影响(7 d),注:种衣剂为克.醇.福美双;
图1、图2中的横坐标“The coaking concentrations of glyphosate”含义为“包衣剂中草甘膦浓度”,纵坐标“The Normal Seedling percentage”含义为“正常苗率”;“Lianchuang505”含义为“联创505”,“Shuangkang12-5”含义为“双抗12-5”,“zhengdan958”含义为“郑单958”;
图3为含草甘膦的种衣剂包衣对双抗12-5、联创505和郑单958幼苗生长的影响,注:图a,b,c所用商品种衣剂为克.醇.福美双;图d,e,f所用商品种衣剂为克.戊.福美双;图g,h,i所用商品种衣剂为噻虫嗪。
具体实施方式
下面结合实施例对本申请做进一步的解释说明,在介绍具体实施例前,就下述实施例中涉及部分实验材料、实验试剂等实验背景情况简要介绍说明如下。
实验材料:
抗草甘膦玉米品种双抗12-5和其相应的遗传背景材料联创505,由浙江大学提供;
普通玉米对照郑单958,属于一种常用玉米栽培品种,购置于秋乐种业有限公司;
实验试剂:
玉米种衣剂克.醇.福美双(克百威+三唑醇+福美双)、克.戊.福美双(克百威+戊唑醇+福美双)和噻虫嗪,均由北农(海利)涿州种衣剂有限公司提供;
41%草甘膦异丙胺盐溶液(农达),美国孟山都公司产品。
实施例1
本实施例所采用草甘膦包衣剂,为含有一定浓度草甘膦的商品化玉米种衣剂克.醇.福美双(克百威+三唑醇+福美双),具体制备时,在工作浓度的种衣剂中加入41%浓度的草甘膦至所需草甘膦终浓度即可。为确定草甘膦包衣剂中草甘膦浓度的合适用量,以种子的出苗率和正常苗率,幼苗的形态指标和叶绿素含量等为参考标准,发明人进行了相关实验,具体实验过程简要介绍如下。
实验过程:
将玉米种子采用包衣剂处理后,在光照培养箱(28℃、16 h光/ 26℃、8 h 暗)中生长培育7 d,进行出苗率、正常苗率、形态指标和叶绿素含量的测量和计算。
每个处理出苗率=(发芽幼苗数量/播种种子数量)×100%;
每个处理正常苗率=(正常幼苗数量/播种种子数量)×100%;
测定完成后,将幼苗从土中取出,用自来水将幼苗根部的土冲洗干净,并用纸巾将植株上的水完全吸干后进行拍照;之后用电子天平称量幼苗全株鲜重和叶重,用不锈钢直尺测量根长和茎长。
叶绿素含量测定:使用便携式叶绿素测定仪SPAD-502 Plus (Minolta CameraCo. Ltd., Osaka, Japan)测量玉米叶片不同部位叶绿素含量,每株测量6-8次,取平均值作为该株的叶绿素含量,每个处理测量5株。
实验分组情况:共分为6个处理组,以种衣剂克.醇.福美双直接包衣的玉米种子为对照(药剂和种子按照1:50的质量比例直接进行包衣);制备本申请中所述草甘膦包衣剂时,在种衣剂克.醇.福美双(种衣剂克.醇.福美双总的用量与对照组相同)中加入41%浓度的草甘膦至草甘膦终浓度分别为0.25%、0.50%、1.00%、2.00%和4.00%五个浓度,作为5个实验处理组。
培养7d后,对玉米发芽率统计结果如图1所示。从图1中可以看出,作为抗草甘膦品种,在0.25~4.00%浓度的草甘膦包衣情况下,双抗12-5发芽率均显著的高于其遗传背景材料联创505和普通玉米品种对照郑单958;而在草甘膦浓度不超过1%情况下,双抗12-5发芽率未见显著变化,只有当草甘膦浓度不低于2.00%时,对于双抗12-5发芽率才造成一定影响。而需要注意的是,在草甘膦浓度不低于2.00%时,不具有草甘膦抗性的联创505和郑单958均不能正常发芽,因此,仅就发芽率指标而言,包衣剂中草甘膦浓度不宜低于2.00%,从而在播种后可以有效去除不具有草甘膦抗性的杂株,以便及时补种和后期管理。
培养7d后,对玉米正常苗率统计结果如图2所示。从图2中可以看出,作为抗草甘膦品种,在0.25~4.00%浓度的草甘膦包衣情况下,双抗12-5正常苗率均显著的高于联创505和郑单958。观察及统计结果表明,联创505和郑单958对包衣剂中所添加草甘膦均十分敏感,在终浓度为0.25%条件下,正常苗率即发生显著的下降,当草甘膦终浓度达到0.50%时,即使已发芽的幼苗,均表现为植株极度矮小、叶片白化,显现出明显的草甘膦危害状况,正常苗率降为0(部分形态图如图3所示);而同等浓度条件下,双抗12-5所有已发芽的幼苗均可正常生长。因此,仅以正常苗率作为考量指标时,包衣剂中草甘膦浓度显然不宜低于0.50%,以有效筛除不具有草甘膦抗性种苗。
将玉米发芽率和正常苗率结果综合分析可知:(1)只有当草甘膦终浓度不低于2.00%时,双抗12-5发芽率才造成一定影响;(2)当草甘膦终浓度达到0.50%时,联创505和郑单958正常苗率均降为0。因此,包衣剂中草甘膦浓度显然不宜超过2.00%,但也不宜低于0.50%,以保证抗草甘膦玉米发芽率不受影响,并可有效筛除不具有草甘膦抗性的种苗。
培养7d后,对不同浓度下玉米幼苗的全株鲜重(幼苗或种子)、叶重、茎长和根长的统计如下表1所示。
表1,草甘膦包衣对双抗12-5、联创505和郑单958幼苗全株鲜重、叶重、茎长和根长的影响(7 d)
注:种衣剂为克.醇.福美双,不同的小写字母表明同一行内的差异达到显著水平。
对上表数据分析可以看出,随着包衣剂中草甘膦浓度的增加,双抗12-5幼苗全株鲜重并未见显著的降低,对草甘膦表现出较强的耐性,而联创505和郑单958对草甘膦较为敏感,随着草甘膦浓度的增大,幼苗全株鲜重均发生显著的下降。而当草甘膦终浓度达到1.00%时,联创505和郑单958中绝大多数种子不能发芽,即使发芽的幼苗均极度矮小。
将幼苗分为根、茎、叶三部分分别进行考察后,分析表明:抗草甘膦玉米品种双抗12-5幼苗的叶重、茎长和根长各项指标均未见显著的降低,而联创505和郑单958随着草甘膦浓度的增大,各项指标均发生显著的下降,与全株鲜重的分析结果表现出相同和相似的规律。
对玉米叶片叶绿素含量测定结果如下表2所示。
表2,草甘膦包衣对双抗12-5、联创505和郑单958幼苗叶片叶绿素含量的影响(7d)
注:种衣剂为克.醇.福美双,不同的小写字母表明同一行内的差异达到显著水平。
从上表数据可以看出,随着草甘膦浓度增大,双抗12-5叶绿素含量在各处理组间未见显著差异;而联创505和郑单958对草甘膦较为敏感,随草甘膦浓度的增加,两者叶片叶绿素含量均发生显著的下降,这与形态指标的考察结果相似。
综合上述实验结果,为较好去除转基因种子中非转基因种子影响,应用包衣剂包裹处理玉米种子时,包衣剂中草甘膦浓度可以在0.5~4%之间使用,综合发芽率和成苗率而言,较优浓度范围为0.5~2%,进一步优选浓度应为1~2%。
实施例2
在实施例1基础上,为进一步检验草甘膦使用浓度是否与种衣剂具有关联性,参考实施例1相关操作,分别以商品化种衣剂克.醇.福美双、克.戊.福美双(将粉剂按照质量比1:4兑水混匀后,再将药剂和种子按照1:50的质量比例进行包衣)和噻虫嗪(按照体积比1:1兑水混匀后,再将药剂和种子按照1:50的质量比例进行包衣)为基础,分别添加41%浓度的草甘膦至终浓度为0.50%和1.00%,相应种衣剂直接作为包衣剂为对照,每个处理包衣40粒玉米种子,3次重复。
对不同浓度处理组正常苗率进行统计,结果如下表3所示。
表3,三种包衣剂对双抗12-5、联创505和郑单958正常苗率的影响 (7 d)
注:不同的小写字母表明同一行内的差异达到显著水平。
对上表数据进行分析,可以看出,双抗12-5正常苗率不受草甘膦浓度和包衣剂类型影响,均能正常成苗。而联创505和郑单958的成苗率则同时受种衣剂和草甘膦浓度影响,具体而言:
使用种衣剂克.醇.福美双包衣时,草甘膦终浓度为0.5%时,联创505和郑单958的正常苗率均为0;
使用种衣剂克.戊.福美双和噻虫嗪包衣时,草甘膦终浓度为0.5%时,郑单958正常苗率均降为0,而联创505正常苗率仍然有25.00%和13.33%,这一方面表明不同玉米品种间对于草甘膦抗性表现出显著的不同,另一方面也说明,这一浓度的草甘膦尚不足以明确区分开抗性品种幼苗和非抗性品种幼苗;而在草甘膦浓度达到1.00%时,联创505和郑单958的正常苗率均降为0。
综上结果可以看出,为有效区分草甘膦抗性品种和非抗性品种,应用包衣剂包衣处理玉米时,种衣剂中草甘膦浓度不宜低于1.00%,从而尽量减少非抗性品种中品种间差异及种衣剂差异影响。
Claims (5)
1.一种抗草甘膦玉米用草甘膦包衣剂,其特征在于,所述包衣剂为含有草甘膦的种衣剂,所述种衣剂为克.醇.福美双、克.戊.福美双或噻虫嗪,所述包衣剂中草甘膦质量浓度为0.5~4%。
2.如权利要求1所述抗草甘膦玉米用草甘膦包衣剂,其特征在于,包衣剂中草甘膦质量浓度为0.5~2.0%。
3.如权利要求2所述抗草甘膦玉米用草甘膦包衣剂,其特征在于,包衣剂中草甘膦质量浓度为1.0~2.0%。
4.权利要求1~3任一项所述抗草甘膦玉米用草甘膦包衣剂在玉米栽培中的应用,其特征在于,用于具有草甘膦抗性的玉米品种。
5.如权利要求4所述抗草甘膦玉米用草甘膦包衣剂在玉米栽培中的应用,其特征在于,所述玉米品种为抗草甘膦的双抗12-5。
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CN104126594A (zh) * | 2014-08-02 | 2014-11-05 | 山东罗邦生物农药有限公司 | 一种悬浮种衣剂生产工艺 |
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