An improved method to efficiently acquire rice black-streaked dwarf virus viruliferous small brown planthoppers

Linlin Du; Bo Zeng; Xuejuan Li; Ying Lan; Wei Guo; Zhaoyun Wang; Zhiyang Liu; Yijun Zhou; Kumar Kunda Jiban; Tong Zhou

doi:10.3389/fgene.2023.1111030

An improved method to efficiently acquire rice black-streaked dwarf virus viruliferous small brown planthoppers

Front Genet. 2023 Feb 1:14:1111030. doi: 10.3389/fgene.2023.1111030. eCollection 2023.

Authors

Linlin Du^{1

2}, Bo Zeng³, Xuejuan Li², Ying Lan², Wei Guo², Zhaoyun Wang², Zhiyang Liu², Yijun Zhou², Kumar Kunda Jiban⁴, Tong Zhou^{1

2}

Affiliations

¹ 1 College of Plant Protection, Nanjing Agricultural University, Nanjing, China.
² Jiangsu Key Laboratory for Food Quality and Safety-State Key Laboratory Cultivation Base of Ministry of Science and Technology/International Rice Research Institute and Jiangsu Academy of Agricultural Sciences Joint Laboratory/Institute of Plant Protection, Jiangsu Academy of Agricultural Sciences, Nanjing, China.
³ National Agricultural Technology Extension Service Center, Beijing, China.
⁴ Plant Virus and Vector Interactions, Centre for Plant Virus Research, Crop Research Institute, Prague, Czechia.

Abstract

Accurate phenotypic identification is the basis of research for resistance genetics and rice breeding for resistance to RBSDV disease. Obtaining rice black-streaked dwarf virus (RBSDV) viruliferous small brown planthoppers (SBPHs) with high transmission efficiency is an essential part of accurate phenotypic identification. Here, through quantifying number of RBSDV copies in infected rice plants, optimizing times of SBPHs fed on RBSDV-infected rice plants and leaf stage of rice seedlings, a method to acquire an RBSDV-carrying SBPH population more efficiently was improved. The results showed that rate of viruliferous SBPHs was significantly higher when fed on RBSDV-infected rice plants that had the copy numbers of RBSDV S10 of 3.0*10⁴ and 1.1*10⁴ than 8.3*10². Therefore, it is more efficient for SBPHs to acquire the virus when fed on RBSDV-infected rice plants that have copy numbers of RBSDV S10 above 1.1*10⁴. The rate of viruliferous SBPHs were 50% and 54%, respectively, after the insects fed on RBSDV-infected rice plants for 7 and 9 days and being transferred to healthy rice seedlings for 5 and 3 days, which was significantly higher than those at other feeding times. The optimal inoculation leaf stage of rice seedlings was the 2-3-leaf stage (3 effective SBPHs per seedling for 72 h), but a high rate of viruliferous SBPHs may be suggested for inoculation of older rice seedlings.

Keywords: RBSDV; RT-qPCR; feeding times; viruliferous SBPHs; virus loads.

Grants and funding

This study was supported by the National Natural Science Foundation of China (Grant No. 31901959), the Open Competition Mechanism to Select the Best Candidates Fund of Jiangsu Province (Grant No. JBGS [2021]008), the Key Research and Development Program of Jiangsu Province (Grant No. BE2022369), and the Agricultural Science and Technology Independent Innovation Fund of Jiangsu Province (Grant No. CX [22] 3019). JK was supported by the Ministry of Agriculture of the Czech Republic (Grant No. Mze RO 0418).