番茄NP24基因的遗传转化

本研究旨在通过将类甜蛋白基因NP24 导入番茄，提高番茄品种的抗病能力。通过对‘京乐502’、‘耐运2000’以及‘Bolgogragsky’3 个番茄品种进行抗病性的鉴定及防御酶活性测定，选取抗病性最弱的番茄品种‘Bolgogragsky’，采用农杆菌介导法将类甜蛋白基因NP24 导入番茄。接种晚疫病7 天后测得‘Bolgogragsky’、‘耐运2000’和‘京乐502’病情指数分别为81.7、48.7、16.0。‘Bolgogragsky’POD酶活性在第1 天达到高峰，‘京乐502’、‘耐运2000’在第5 天到达最高峰，但‘京乐502’的酶活性一直较其他2 组高。PAL酶活性先升高后降低，‘京乐502’升得最快降得最慢，‘Bolgogragsky’升得最慢降得最快。‘京乐502’SOD酶活性在0~5 天升高而后降低，‘耐运2000’、‘Bolgogragsky’接种后第1 天SOD酶活性明显降低，后续第3、5、7 天在波动中上升。实验结果表明接种晚疫病菌后3 个品种番茄的POD、PAL酶活性均高于对照，且酶活性与病情指数呈负相关；而处理组SOD酶活性均较对照组有所降低。此外，经PCR验证NP24基因已经整合到番茄‘Bolgogragsky’基因组中。

Genetic Transformation of NP24 Gene in Tomato

The study aims at improving the disease resistance of tomato varieties by inducing haumatin-like proteins gene NP24 into tomato. Through the identification of disease resistance and the activity of defensive enzyme of‘Jingle 502’,‘Naiyun 2000’and‘Bolgogragsky’, the weakest disease- resistance variety ‘Bolgogragsky’was selected to transform the NP24 gene with the Agrobacterium- mediated method. The disease index of‘Bolgogragsky’,‘Naiyun 2000’and‘Jingle 502’after 7 days of Phytophthora infestans inoculation was 81.7, 48.7 and 16.0, respectively. The activity of POD enzyme in‘Bolgogragsky’reached the peak on the first day, while that of‘Naiyun 2000’and‘Jingle 502’peaked on the 5th day, but‘Jingle 502’ had consistently higher enzyme activity than that of the other two varieties. The activity of PAL enzyme increased first and then decreased,‘Jingle 502’showed the fastest increase and the slowest decrease. ‘Bolgogragsky’showed the lowest increase and the fastest decrease. The activity of SOD enzyme in‘Jingle 502’increased on the 0-5th day and then decreased, but the activity of SOD enzyme in‘Bolgogragsky’and ‘Naiyun 2000’significantly decreased on the first day after inoculation, and displayed an increase with fluctuation on the 3rd, 5th and 7th day. The results indicated that after inoculation of Phytophthora infestans, the enzyme activities of POD and PAL in the three tomato varieties were higher than that of the control, and the enzyme activity had a negative correlation with the disease index; nevertheless, the enzyme activity of SOD in treated groups was lower than that of the control. Moreover, PCR analysis approved that NP24 gene had been transformed into the tomato genome of‘Bolgogragsky’