江苏农业学报
江囌農業學報
강소농업학보
JIANGSU JOURNAL OF AGRICULTURAL SCIENCES
2014年
5期
1102-1108
,共7页
杨玛丽%张保龙%郭佳茹%赵统敏%余文贵%赵丽萍%王银磊
楊瑪麗%張保龍%郭佳茹%趙統敏%餘文貴%趙麗萍%王銀磊
양마려%장보룡%곽가여%조통민%여문귀%조려평%왕은뢰
番茄黄化曲叶病毒%RNAi%GroEL%共转化%转基因番茄
番茄黃化麯葉病毒%RNAi%GroEL%共轉化%轉基因番茄
번가황화곡협병독%RNAi%GroEL%공전화%전기인번가
tomato yellow leaf curl virus (TYLCV)%RNAi%GroEL%Co-transformation%transgenic tomato
本研究将已构建的SUC2-GroEL和pBI121-AC1-AC2植物表达载体共转化番茄,共获得19株卡那霉素抗性再生植株。经过PCR和RT-PCR检测,初步获得转SUC2-GroEL载体的阳性植株5株,转pBI121-AC1-AC2载体的阳性植株3株以及转SUC2-GroEL/ pBI121-AC1-AC2双载体的阳性植株2株。利用TYLCV侵染性克隆农杆菌注射接种法对T0代转基因阳性植株进行抗性鉴定,获得抗TYLCV的转基因植株7株。对T0代未扩出病毒条带的T1代植株进行抗性鉴定,结果发现转SUC2-GroEL、pBI121-AC1-AC2、SUC2-GroEL/pBI121-AC1-AC2不同载体的株系植株中平均带毒率依次是35%、25%和15%,推断转双基因的植株的抗病能力优于转单个基因植株的抗病能力。
本研究將已構建的SUC2-GroEL和pBI121-AC1-AC2植物錶達載體共轉化番茄,共穫得19株卡那黴素抗性再生植株。經過PCR和RT-PCR檢測,初步穫得轉SUC2-GroEL載體的暘性植株5株,轉pBI121-AC1-AC2載體的暘性植株3株以及轉SUC2-GroEL/ pBI121-AC1-AC2雙載體的暘性植株2株。利用TYLCV侵染性剋隆農桿菌註射接種法對T0代轉基因暘性植株進行抗性鑒定,穫得抗TYLCV的轉基因植株7株。對T0代未擴齣病毒條帶的T1代植株進行抗性鑒定,結果髮現轉SUC2-GroEL、pBI121-AC1-AC2、SUC2-GroEL/pBI121-AC1-AC2不同載體的株繫植株中平均帶毒率依次是35%、25%和15%,推斷轉雙基因的植株的抗病能力優于轉單箇基因植株的抗病能力。
본연구장이구건적SUC2-GroEL화pBI121-AC1-AC2식물표체재체공전화번가,공획득19주잡나매소항성재생식주。경과PCR화RT-PCR검측,초보획득전SUC2-GroEL재체적양성식주5주,전pBI121-AC1-AC2재체적양성식주3주이급전SUC2-GroEL/ pBI121-AC1-AC2쌍재체적양성식주2주。이용TYLCV침염성극륭농간균주사접충법대T0대전기인양성식주진행항성감정,획득항TYLCV적전기인식주7주。대T0대미확출병독조대적T1대식주진행항성감정,결과발현전SUC2-GroEL、pBI121-AC1-AC2、SUC2-GroEL/pBI121-AC1-AC2불동재체적주계식주중평균대독솔의차시35%、25%화15%,추단전쌍기인적식주적항병능력우우전단개기인식주적항병능력。
Tomato yellow leaf curl virus ( TYLCV) is one of the most devastating viruses of cultivated tomatoes in the world. Recently, the disease occurred suddenly in China, spreading towards north from south, which has resulted in great economic losses. The lack of resistance resource limits the TYLCV resistance breeding. In order to generate engineered re-sistance, the plant expression vectors SUC2-GroEL and pBI121-AC1-AC2 were co-transformed into tomato by agrobacterium-mediated genetic transformation method, and nineteen kanamycin resistant regenerated tomato plants were obtained, inclu-ding five plants of SUC2-GroEL vector, three plants of pBI121-AC1-AC2 vector and three plants of double vectors confirmed by PCR and RT-PCR. T0 generation of the transgenic plants were inoculated with TYLCV, and it turned out 8 were TYLCV resistant. The carrying rates of TYLCV by T1 generation achieved via vectors SUC2-GroEL, pBI121-AC1-AC2, and SUC2-GroEL/pBI121-AC1-AC2 were 35%, 25%, and 15%, respectively, suggesting that the plants with double genes have a stronger disease resistance than other plants with single gene.