CAJ | 학술논문

TILLING(Targeting induced local lesions in genomes,定向诱导基因组局部突变技术)是一种高通量的等位变异创制和突变体快速鉴定技术,其实质是将传统的化学诱变方法和突变的高效筛选有效结合的反向遗传学研究方法.其技术原理是将传统的酶切技术与PCR技术相结合后采用红外双色荧光系统进行结果鉴定,从而筛选出相应的突变体.传统的TILLING技术主要用于筛选由人工诱导产生的突变体.Ecotilling技术由TILLING技术延伸而来,主要用于鉴定自然界中已经存在的突变体,其与传统的TILLING技术的区别主要为构建DNA池时略有差异.随着该项技术在拟南芥等模式植物中的成功应用,越来越多的人开始将其用于基因组较大的植物之中.本文对近年来TILLING技术在麦类作物中的应用进行了分析,并通过比较不同植物突变体库中的突变频率发现,经EMS处理的小麦等麦类作物突变体库中的突变频率显著高于其他植物,因此相信,TILLING技术将会作为一种常规手段在麦类作物尤其是普通小麦改良中得到越来越广泛的应用.
TILLING(Targeting induced local lesions in genomes,정향유도기인조국부돌변기술)시일충고통량적등위변이창제화돌변체쾌속감정기술,기실질시장전통적화학유변방법화돌변적고효사선유효결합적반향유전학연구방법.기기술원리시장전통적매절기술여PCR기술상결합후채용홍외쌍색형광계통진행결과감정,종이사선출상응적돌변체.전통적TILLING기술주요용우사선유인공유도산생적돌변체.Ecotilling기술유TILLING기술연신이래,주요용우감정자연계중이경존재적돌변체,기여전통적TILLING기술적구별주요위구건DNA지시략유차이.수착해항기술재의남개등모식식물중적성공응용,월래월다적인개시장기용우기인조교대적식물지중.본문대근년래TILLING기술재맥류작물중적응용진행료분석,병통과비교불동식물돌변체고중적돌변빈솔발현,경EMS처리적소맥등맥류작물돌변체고중적돌변빈솔현저고우기타식물,인차상신,TILLING기술장회작위일충상규수단재맥류작물우기시보통소맥개량중득도월래월엄범적응용.
Targeting Induced Local Lesion in Genomes (TILLING) is a reverse genetic strategy that uses traditional chemical mutagenesis methods to create libraries of mutagenized individuals later subjected to high throughput screens for the discovery of an allelic series of induced point mutations in target genes. Usually, TILLING technique consisted of applications of restriction enzyme digestion and Polymerase Chain Reaction (PCR) as well as infrared fluorescence detection of IRD700 nm and IRD 800 nm on acrylamide electrophresis by Licor-segue genetic analyser. Differing from TILLING only for induced mutations by chemical mutagens, Ecotilling stemming from traditional TILLING is generally used for identification of natural mutations and an obvious difference between TILLING and Ecotilling is the mean of establishing DNA pools. With the successful application of TILLING and Ecotilling in model plants such as Arabidopsis thaliana ect., more and more studies are focusing on the improvement of plants with large genome size. In this review, application of TILLING in Triticeae crops was reviewed and compared with sharply higher percentage of mutation than those of other reported plants. Therefore, TILLING will be promisingly used as a common method to improve characters of Triticeae crops, especially bread wheat (Triticum eastivum L.).