Abstract:The public concern about pollen-mediated gene flow (PGF) from genetically modified (GM) crops to non-GM crops heats up in recent years over China. In the current study, we conducted greenhouse and field experiments to measure PGF with various physical isolation measures, including 90, 80, 60 and 40 holes/cm2 separation nets and Sorghum bicolor, Zea mays and Lycopersicon esculentum separation crops between GM cotton and non-GM line (Shiyuan321) by seed DNA test during 2013 to 2015, and pollen grain dyeing was a… Show more
“…Transgenic plants can be engineered to express hairpin RNAs (hpRNAs) against some functional genes in chewing and piercingsucking pests (Pitino et al, 2011;Zha et al, 2011;Thakur et al, 2014;Zhang et al, 2017). However, the major obstacle may come from the potential ecological consequences and public concerns of genetically modi ed foods (Yan et al, 2015(Yan et al, , 2018(Yan et al, , 2020a. Meanwhile, there are also promising non-transformative RNAi applications/approaches available.…”
There are two main limitations for sprayable RNA pesticide development: delivery e ciency and synthetic cost of double-stranded RNA (dsRNA). We previously constructed a nanocarrier-based transdermal dsRNA delivery system and a novel bacteria-based hairpin RNA (hpRNA) expression system to solve these challenges. Herein, as a subsequent exploration of RNA pesticide (sprayable ds/hpRNA for pest control), we performed a greenhouse application of bacteria-expressed and nanocarrier-delivered RNA pesticide on green peach aphid. The nanoparticle SPc could combine and deliver dsRNA across the aphid cuticle, and V-type proton ATPase subunits d (ATP-d) and G (ATP-G) were selected as the potential RNA interference (RNAi) targets. Our plasmid-Escherichia coli system simultaneously expressing ATP-d and ATP-G hairpin RNAs (hpRNAs) was constructed for mass production of hpRNA. The expressed hpRNA was mixed with SPc and detergent to form RNA formulation, which showed a certain insecticidal activity through the spray application in the greenhouse. Total control e cacy of our RNA pesticide could reach 61% on 3 d, and maintained at 50% until the sixth day. To our knowledge, our study is the rst attempt to apply the bacteria-expressed and nanocarrier-delivered RNA pesticides for pest control in the greenhouse trial, which is bene cial for promoting the development of RNA pesticides.
Key Message• SPc can protect and deliver dsRNA from degradation for e cient gene silencing• hpRNA is expressed in E. coli system to develop a novel RNA insecticide • Our RNA pesticide can be applied to control aphids in greenhouse • Our study brings the concept of RNAi-based pest management to practical application
“…Transgenic plants can be engineered to express hairpin RNAs (hpRNAs) against some functional genes in chewing and piercingsucking pests (Pitino et al, 2011;Zha et al, 2011;Thakur et al, 2014;Zhang et al, 2017). However, the major obstacle may come from the potential ecological consequences and public concerns of genetically modi ed foods (Yan et al, 2015(Yan et al, , 2018(Yan et al, , 2020a. Meanwhile, there are also promising non-transformative RNAi applications/approaches available.…”
There are two main limitations for sprayable RNA pesticide development: delivery e ciency and synthetic cost of double-stranded RNA (dsRNA). We previously constructed a nanocarrier-based transdermal dsRNA delivery system and a novel bacteria-based hairpin RNA (hpRNA) expression system to solve these challenges. Herein, as a subsequent exploration of RNA pesticide (sprayable ds/hpRNA for pest control), we performed a greenhouse application of bacteria-expressed and nanocarrier-delivered RNA pesticide on green peach aphid. The nanoparticle SPc could combine and deliver dsRNA across the aphid cuticle, and V-type proton ATPase subunits d (ATP-d) and G (ATP-G) were selected as the potential RNA interference (RNAi) targets. Our plasmid-Escherichia coli system simultaneously expressing ATP-d and ATP-G hairpin RNAs (hpRNAs) was constructed for mass production of hpRNA. The expressed hpRNA was mixed with SPc and detergent to form RNA formulation, which showed a certain insecticidal activity through the spray application in the greenhouse. Total control e cacy of our RNA pesticide could reach 61% on 3 d, and maintained at 50% until the sixth day. To our knowledge, our study is the rst attempt to apply the bacteria-expressed and nanocarrier-delivered RNA pesticides for pest control in the greenhouse trial, which is bene cial for promoting the development of RNA pesticides.
Key Message• SPc can protect and deliver dsRNA from degradation for e cient gene silencing• hpRNA is expressed in E. coli system to develop a novel RNA insecticide • Our RNA pesticide can be applied to control aphids in greenhouse • Our study brings the concept of RNAi-based pest management to practical application
“…The most effective means to reduce outcrossing and PMGF in GM triticale may be developing cultivars that exhibit a lower propensity for floret opening [12]. Additionally, barriers at field borders, such as tall, dense plants or nets are highly desirable [50].…”
A transgenic winter triticale line expressing the uidA gene, encoding β-glucuronidase, was used to assess the pollen flow in field experiments over two consecutive vegetation seasons in central Poland. The experimental design included two variants of mixed transgenic and non-transgenic lines. Pollen grains were collected using passive traps located at 0, 10, 30, 60 and 85 meters from the transgenic line. GM pollen grains were detected histochemically by staining with x-Gluc. A positive effect of temperature increase, as well as the strength and direction of the wind on the number and spread of pollen grains was observed. Regardless of the experiment year and variant, only few pollen grains were observed at a distance of 85 m. In the first year of the study the amount of pollen grains at 85 m was 300-fold lower than at the source and 140-fold lower in the second year. The number of transgenic pollen grains was two times lower when the field with the transgenic triticale was surrounded by a non-transgenic line, compared to an empty field. On the basis of the obtained results, we suggest 100 m as the distance for triticale pollen migration, although longer flight incidents are possible in extreme atmospheric conditions.
“…However, chloroplast transformation protocols still need to be developed for cereals such as rice, wheat, and maize to expand the crop range of transplastomic technology (Zhang et al, 2017). Additionally, transplastomic crops are still considered transgenic products in most countries, requiring them to undergo a rigorous evaluation because of their potential environmental risk (Andow & Zwahlen, 2006;Yan et al, 2018Yan et al, , 2020a.…”
RNA interference (RNAi) targeting lethal genes in insects has great potential for sustainable crop protection. Compared with traditional double-stranded (ds)RNA delivery systems, nanoparticles such as chitosan, liposomes, and cationic dendrimers offer advantages in delivering dsRNA/small interfering (si)RNA to improve RNAi efficiency, thus promoting the development and practice of RNAi-based pest management strategies. Here, we illustrate the limitations of traditional dsRNA delivery systems, reveal the mechanism of nanoparticle-mediated RNAi, summarize the recent progress and successful applications of nanoparticle-mediated RNAi in pest management, and finally address the prospects of nanoparticle-based RNA pesticides.
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