Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (10): 1710-1720.doi: 10.3864/j.issn.0578-1752.2019.10.005

• PLANT PROTECTION • Previous Articles     Next Articles

Disease Resistance of Rice stripe virus NS3-Transgenic Nicotiana benthamiana

WU GenTu,CHEN GuangXiang,ZHANG JiaYuan,HU Qiao,MA MingGe,DOU YanXia,LI MingJun,QING Ling()   

  1. Chongqing Key Laboratory of Plant Disease Biology, College of Plant Protection, Southwest University, Chongqing 400716
  • Received:2019-01-15 Accepted:2019-03-04 Online:2019-05-16 Published:2019-05-23
  • Contact: Ling QING E-mail:qling@swu.edu.cn

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Abstract:

【Objective】The protein encoded by NS3 is a suppressor of RNA silencing of Rice stripe virus (RSV). In previous study, it was found that NS3 expression enhanced the resistance of Nicotiana benthamiana to RSV using transgenic technology. In order to reveal the detailed function of NS3 in host plants, the resistance of NS3-transgenic N. benthamiana to other tobacco diseases will be analyzed in this study.【Method】Wild-type and NS3-transgenic N. benthamiana were used to study the infection of Potato virus X (PVX), Ralstonia solanacearum, and Phytophthora parasitica var. nicotianae. PVX infectious clone was inoculated by agro-infiltration, viral symptoms were observed. The total RNA of infected leaves was extracted, and viral relative accumulation was detected by RT-qPCR. The method of filling root was used to inoculate R. solanacearum and P. parasitica var. nicotianae, respectively, disease symptoms were observed, and the incidence and disease index were analyzed.【Result】The main symptoms of PVX in N. benthamiana leaves were mosaic and chlorosis, and the NS3-transgenic N. benthamiana plants had the same PVX symptoms as those of wild-type plants. At 10 days past inoculation (dpi), the results of RT-qPCR showed that NS3 expression inhibited the accumulation of PVX in N. benthamiana, the relative accumulation of PVX in NS3-6 line and NS3-9 line was 68.17% and 81.01% of that of wild-type N. benthamiana, respectively. After the infection of R. solanacearum, the symptoms both in wild-type and NS3-transgenic N. benthamiana plants were wilting and damping-off. In the early stage of disease, the expression of NS3 was beneficial to the infection of R. solanacearum to a certain extent, which showed that it was more sensitivity to bacterial wilt. At 14 dpi, the incidence and disease index of bacterial wilt of wild-type N. benthamiana were 100.00% and 78.67, while the incidence of NS3-6, NS3-9 line was 95.00%, 98.33%, and the disease index was 70.00, 73.00, respectively. After 14 dpi, the incidence and disease index increased gradually, while the disease index of NS3-transgenic lines was lower than that of wild-type plants. The black-brown necrotic patches at the base of stem were observed both in wild-type and transgenic plants after P. parasitica var. nicotianae infection, and the expression of NS3 did not affect the symptoms of N. benthamiana. At the early stage of P. parasitica var. nicotianae infection, the incidence of black shank disease in NS3-6 was higher than that of wild-type N. benthamiana, however, during the whole process of black shank disease, the disease index of NS3-transgenic lines was lower than that of wild-type N. benthamiana. At 12 dpi, the incidence and disease index of black shank disease in wild-type N. benthamiana were 96.67% and 77.50, while the incidence of NS3-6, NS3-9 line was 90.00%, 86.67%, and the disease index was 64.17, 62.08, respectively.【Conclusion】The observation of pathogen infection process showed that the expression of NS3 affected the infection and severity of tobacco diseases on N. benthamiana to a certain extent, and the effects on different pathogens were also different.

Key words: Rice stripe virus (RSV), NS3, transgenic Nicotiana benthamiana, disease resistance

Fig. 1

The identification of NS3-transgenic N. benthamiana by PCR"

Fig. 2

The symptom of PVX-infecting N. benthamiana"

Fig. 3

The relative accumulation level of PVX in N. Benthamiana Each column represents the average of three independent experiments. * indicates significant difference (0.01≤ P<0.05). The same as below"

Fig. 4

The symptom of different disease grades of N. benthamiana plants infected with R. solanacearum"

Fig. 5

The incidence of N. benthamiana after R. solanacearum infection"

Fig. 6

The disease index of N. benthamiana after R. solanacearum infection"

Fig. 7

The symptom of different disease grades of N. benthamiana infected with P. parasitica var. nicotianae"

Fig. 8

The incidence of N. benthamiana after P. parasitica var. nicotianae infection"

Fig. 9

The disease index of N. benthamiana after P. parasitica var. nicotianae infection"

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