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Production of Herbicide-Resistant Crops by Cell-Engineering

通过细胞工程生产抗除草剂作物

基本信息

批准号：
05660057
负责人：
MATSUNAKA Shooichi
金额：
$ 1.34万
依托单位：
Kansai University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (C)
财政年份：
1993
资助国家：
日本
起止时间：
1993 至 1995
项目状态：
已结题

项目摘要

Long time and much money are required for the development of a new selective herbicide for a crop plant. If we can create a crop plant having resistance to a herbicide which is low-toxic to animals, safe to the environment, cheap and has a long practical use history, it is evaluated as the same matter as the development of above-described new selective herbicides for such crop plant.In this study, severaltrials were examined, and as the examples having some possibility for the research purpose, the creation of glyphosate-resistant bent putter and paraquat-resistant sunflower.In the selection of glyphsate-resistant bent butter, callus having resistance to 1,000 uM glyphosate were obtained by the gradual increasing of the concentration of co-existing herbicide from 100 until 1,000 uM.This tolerant callus was transplanted to hormone-free medium and cultured in the light. After 20 days, the generated plants were soaked in 1,000 uM glyphosate solution for 8 hours. After more 20 days the plant regenerated from resistant callus were survival while ones generated from susceptible callus were dead. The sikimic acid content of the resistant one was one sixth of original susceptible one.On the other hand, both protoplasts prepared from leaves of sunflower and paraquat-resistant horseweed by enzyme treatment were fused by a electrofusion system (SHIMAZU SOMATIC HYBRIDIZER,SSH-10). The concentration of both protoplasts was 100 thousands cells per ml. The fusates were cultured on MS medium (NAA 0.2 ppm, BA 0.5 ppm), at 25ﾟC under 3,000 lux, for 2 months and moved to paraquat-containing MS medium. Unfused sunflower colonies could not form any callus upper 1 x 10^<-5>M paraquat, under which condition the fused cells could form callus and regenerated plants.

开发一种新的作物选择性除草剂需要很长的时间和大量的资金。如果我们能够创造出对除草剂具有抗性的作物，该除草剂对动物低毒，对环境安全，廉价并且具有长期的实际使用历史，则其被评价为与开发用于这种作物的上述新的选择性除草剂相同的问题。在本研究中，检查了几个试验，并且作为对于研究目的具有一定可能性的实例，在抗草甘膦的弯形黄油的选择中，通过将共存除草剂的浓度从100逐渐增加至1，将该耐受性愈伤组织移植到无蔗糖培养基中并在光照下培养。20天后，将产生的植物浸泡在1，000 μ M草甘膦溶液中8小时。抗性愈伤组织再生的植株经过20天以上仍能存活，而感病愈伤组织再生的植株则死亡。另一方面，利用电融合系统（SHIMAZU SOMATIC HYBRIDIZER，SSH-10）对向日葵和百草枯抗性牛筋草的原生质体进行融合。两种原生质体的浓度均为10万个细胞/ml。融合子在MS培养基（NAA0.2ppm，BA0.5ppm）上，在25 ℃，3,000勒克斯下培养2个月，然后转移到含百草枯的MS培养基上。未融合的向日葵菌落在1 × 10^ M百草枯浓度以上不能形成任何愈伤组织<-5>，在此条件下融合细胞可以形成愈伤组织并再生植株。