Scientia Agricultura Sinica ›› 2019, Vol. 52 ›› Issue (1): 45-55.doi: 10.3864/j.issn.0578-1752.2019.01.005

• PLANT PROTECTION • Previous Articles     Next Articles

Effects of the Interference of Key Magnetic Response Genes on the Longevity of Brown Planthopper (Nilaparvata lugens) Under Near-Zero Magnetic Field

HE JingLan1(),ZHANG Ming1,LIU RuiYing1,WAN GuiJun1,PAN WeiDong2,CHEN FaJun1()   

  1. 1College of Plant Protection, Nanjing Agricultural University, Nanjing 210095
    2 Beijing Key Laboratory of Bioelectromagetics, Institute of Electrical Engineering, Chinese Academy of Sciences, Beijing 100190
  • Received:2018-08-02 Accepted:2018-09-11 Online:2019-01-01 Published:2019-01-12
  • Contact: FaJun CHEN E-mail:2016102054@njau.edu.cn;fajunchen@njau.edu.cn

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Abstract:

【Objective】Cryptochrome (Cry) and iron-sulfur cluster protein IscA (iron-sulfur cluster assembly, MagR) are potential magnetic receptor proteins in organisms. In this study, key magnetic response genes of the brown planthopper (Nilaparvata lugens) were knocked-down by RNA interference (RNAi), including NlCry1, NlCry2 and NlMagR. The objective of this study is to investigate the role of these three magnetic response genes in the longevity mediation of N. lugens in near-zero magnetic field (NZMF). Thus, the response of these three genes to magnetic field could be studied indirectly.【Method】Newly emerged brachypterous female and male adults of N. lugens fed in the lab magnetic field were chosen as the experimental material, and RNAi technology was used to inhibit the key magnetic response genes’ (NlCry1, NlCry2 and NlMagR) expression by injection of double stranded RNA, respectively. Then the RNAi treated adults were immediately transformed into the geomagnetic field (GMF) and NZMF respectively to observe their longevity. The total RNA of the RNAi treated adults under GMF was extracted by using the RNAiso Plus method on the 1st, 2nd and 3rd day after the microinjection, respectively. And then the gene expressions of NlCry1, NlCry2 and NlMagR were measured by using the RT-qPCR (real-time quantitative polymerase chain reaction) after the reverse transcription synthesis of first strand DNA in order to test the efficiency of RNAi. 【Result】There was no significant difference in the longevity of female and male adults after the injection of dsNlCry1 between the treatments of NZMF and GMF, while after the injection of dsNlCry2, the longevity of female (27.78%) and male (50.04%) adults under NZMF was significantly longer than that of the individuals under GMF, respectively. Moreover, the longevity of female adults injected with dsNlCry2 was shorter under GMF while longer under NZMF than that of individuals injected with dsGFP, even if the difference was not significant. The longevity of male adults injected with dsNlCry2 was shorter than that of individuals injected with dsGFP under NZMF (25.41%) and GMF (10.73%), respectively, and the difference under GMF reached the significant level. Furthermore, the longevity of female adults injected with dsNlMagR was significantly shorter (16.48%) than that of individuals injected with dsGFP under the NZMF. 【Conclusion】There is a difference in the regulation of the key genes of magnetic susceptibility (NlCry1, NlCry2 and NlMagR) on the female and male longevity for N. lugens under the change of magnetific field. Hereinto, the NlCry2 susceptibly responses to the changes of magnetic fields, which shows that the gene knock-down and its interaction with magnetic field changes can significantly influence the longevity of female and male adults, and characterized as “sexual dimorphism”. Similarly, the NlMagR (IscA) also sensitively responds to magnetic field changes, but just for the female adults of N. lugens under the NZMF in contrast to the GMF. However, there is no response of NlCry1 to magnetic field changes, and this gene may not be involved in the regulation of female and male longevity for N. lugens.

Key words: magnetic field changes, brown planthopper (Nilaparvata lugens), magnetic response gene, RNA interference (RNAi), adult longevity, magnetic bio-effect

Table 1

Primers and their sequences used in this study"

引物用途 Use of primers 引物名称 Primer name 引物序列 Primer sequence (5′-3′)
cDNA克隆
cDNA cloning		 dsNlCry1-F TAATACGACTCACTATAGGGTCAGACATGGGCTTCGATT
dsNlCry1-R TAATACGACTCACTATAGGGCATTTGGTAAGTTAGCGGTGGA
dsNlCry2-F TAATACGACTCACTATAGGGTGTCAGCATCAATAAGTGGAGG
dsNlCry2-R TAATACGACTCACTATAGGGGCACACCAAACTTGTCGTC
dsNlMagR-F TAATACGACTCACTATAGGGAGAAAGGAAAGTTTGACGAAG
dsNlMagR-R TAATACGACTCACTATAGGGAGCCCTAAATATTAACATCGT
dsGFP-F TAATACGACTCACTATAGGGACGTAAACGGCCACAAGTTC
dsGFP-R TAATACGACTCACTATAGGGTGTTCTGCTGGTAGTGGTCG
实时荧光定量PCR
RT-qPCR		 qNlCry1-F CAGACATGGGCTTCGATTTCA
qNlCry1-R ACCAGCACTTTCTCCGTCAAAT
qNlCry2-F CGCATACTCTCTACAGACTTGAT
qNlCry2-R CACCGTCTGGAATTTGCGATAC
qNlMagR-F CGTTTAATACCTTCAAGAGCAGCAC
qNlMagR-R CCCTACTTTCAAGCCGATAGCAT
qActin-F CTTCTAAACGCCAACCACTCC
qActin-R TCACCCGAAATCACTCACGA
q18S-F TGTCTGCTTAATTGCGATAACGAAC
q18S-R CCTCAAACTTCCATCGGCTTG

Fig. 1

Relative expression level of corresponding RNA of the newly emerged brachypterous female adults of N. lugens after respectively injected with dsNlCry1, dsNlCry2 and dsNlMagR All of the sampled brown planthoppers are newly emerged brachypterous female adults and placed in the geomagnetic field that B=49 μT; * and ** mean significantly different between the dsRNA and the control dsGFP by the T test at P<0.05 and P<0.01, respectively"

Table 2

Two-way ANOVA for the effects of RNAi (dsNlCry1 vs. dsGFP) and magnetic fields (GMF vs. NZMF) on the longevity of female and male adults of N. lugens"

影响因子
Impact factor		 雌虫寿命 Female longevity 雄虫寿命 Male longevity
F值 F value P值 P value F值 F value P值 P value
RNAi 2.87 0.092 0.90 0.34
磁场 Magnetic?field (MF) 0.34 0.56 0.60 0.44
RNAi×磁场 RNAi×MF 0.54 0.47 1.88 0.17

Fig. 2

The longevity of female (A) and male (B) adults of N. lugens after dsNlCry1 injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF) The same lowercase and uppercase letters indicate no significant difference between the treatments of dsNlCry1 and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlCry1 or dsGFP by the T test (P>0.05), respectively"

Table 3

Two-way ANOVA for the effects of RNAi (dsNlCry2 vs. dsGFP) and magnetic fields (GMF vs. NZMF) on the longevity of female and male adults of N. lugens"

影响因子
Impact factor		 雌虫寿命 Female longevity 雄虫寿命 Male longevity
F值 F value P值 P value F值 F value P值 P value
RNAi 2.33 0.13 1.64 0.20
磁场 Magnetic?field (MF) 0.70 0.40 4.14 0.044*
RNAi×磁场 RNAi×MF 4.45 0.037* 7.08 0.009**

Fig. 3

The longevity of female (A) and male (B) adults of N. lugens after dsNlCry2 injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF) Different lowercase and uppercase letters indicate significantly different between the treatments of dsNlCry2 and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlCry2 or dsGFP by the T test (P<0.05), respectively"

Table 4

Two-way ANOVA for the effects of RNAi (dsNlMagR vs. dsGFP) and magnetic fields (GMF vs. NZMF) on the longevity of female and male adults of N. lugens"

影响因子
Impact factor		 雌虫寿命 Female longevity 雄虫寿命 Male longevity
F值 F value P值 P value F值 F value P值 P value
RNAi 3.97 0.048* 0.11 0.74
磁场 Magnetic?field (MF) 1.33 0.25 0.08 0.78
RNAi×磁场 RNAi×MF 0.68 0.41 2.59 0.11

Fig. 4

The longevity of female (A) and male (B) adults of N. lugens after dsNlMagR injection under geomagnetic field (GMF) and near-zero magnetic field (NZMF) Different lowercase and uppercase letters indicate significantly different between the treatments of dsNlMagR and dsGFP for the same sex N. lugens under same magnetic field of GMF or NZMF, and between the treatments of GMF and NZMF for same sex N. lugens injected of dsNlMagR or dsGFP by the T test (P<0.05), respectively"

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