Rice planthoppers, the most devastating rice pests, occur in two wing forms: the short-wing form for rapid population growth and long-wing form for long-distance migration, which together create the mechanism for outbreak. Here we show that Ultrabithorax (Ubx) is a key regulator for switching between the long- and short-wing forms of rice planthoppers. Ubx is expressed in both forewing and hindwing pads, which is different from the canonical model of Ubx expression. In brown planthoppers, expression of Ubx (NlUbx) is regulated by nutritional status of the rice host. High-quality young plants induce NlUbx expression leading to the short-wing form; low-quality ripe plants reduce NlUbx expression resulting in long-wing form. We also showed that NlUbx is regulated by the insulin receptors NlInR1 and NlInR2. The default expression of NlInR1 inhibits NlUbx resulting in long-wings, while high-quality hosts induce NlInR2 expression, which represses NlInR1 thus promoting NlUbx expression to produce short-wings.
BACKGROUNDPromoters play a key role in driving insect‐resistant genes during breeding of transgenic plants. In current transgenic procedures for breeding rice resistance to striped stem borer (Chilo suppressalis Walker, SSB), the constitutive promoter is used to drive the insect‐resistant gene. To reduce the burden of constitutive promoters on plant growth, isolation and identification of insect‐inducible promoters are particularly important. However, few promoters are induced specifically by insect feeding.RESULTSWe found rice hydroperoxide lyase gene (OsHPL2) (LOC_Os02g12680) was upregulated after feeding by SSB. We subsequently cloned the promoter of OsHPL2 and analysed its expression pattern using the β‐glucuronidase (GUS) reporter gene. Histochemical assays and quantitative analyses of GUS activity confirmed that P HPL2:GUS was activated by SSB, but did not respond to brown planthopper (Nilaparvata lugens Stål, BPH) infestation, mechanical wounding or phytohormone treatments. A series of 5′ truncated assays were conducted and three positive regulatory regions (−1452 to −1213, −903 to −624, and −376 to −176) induced by SSB infestation were identified. P2R123‐min 35S and P2TR2‐min 35S promoters linked with cry1C of transgenic plants showed the highest levels of Cry1C protein expression and SSB larval mortality.CONCLUSIONWe identified an SSB‐inducible promoter and three positive internal regions. Transgenic rice plants with the OsHPL2 promoter and its positive regions driving cry1C exhibited the expected larvicidal effect on SSB. Our study is the first report of an SSB‐inducible promoter that could be used as a potential resource for breeding insect‐resistant transgenic crops. © 2020 Society of Chemical Industry
Many essential oils have excellent insecticidal activity and have the potential to be developed as alternatives to chemical insecticides for pest control. Previously, we showed that β‐asarone, a major constituent of the essential oil derived from Acorus calamus L. (Acoraceae), has strong pesticidal activity against brown planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae), the most notorious rice pest in Asia. Here, we report the first evaluation of the sublethal and transgenerational effects of β‐asarone on N. lugens. β‐Asarone significantly decreased female longevity, male longevity, fecundity, and hatchability of F0 generation individuals exposed to the LD30 and LD45 of β‐asarone relative to the acetone control. Moreover, compared with acetone, exposure to LD30 and LD45 of β‐asarone significantly shortened the duration of the egg stage, developmental duration of first instars, and female longevity of F1 generation individuals. Furthermore, the intrinsic rate of increase (r), finite rate of increase (λ), and net reproductive rates (R0) of insects treated with LD30 and LD45 of β‐asarone were significantly lower than those of insects treated with acetone. Compared with acetone, the fecundity and hatchability of F1 generation individuals were significantly decreased after exposure to β‐asarone at LD30 and LD45. These findings indicate the negative effects of sublethal doses of β‐asarone on N. lugens and provide novel information on the potential use of β‐asarone as a substitute chemical pesticide.
Insect pests have a great impact on the yield and quality of crops. Insecticide applications are an effective method of pest control, however, they also have adverse effects on the environment. Using insect‐inducible promoters to drive insect‐resistant genes in transgenic crops is a potential sustainable pest management strategy, but insect‐inducible promoters have been rarely reported. In this study, we found rice allene oxide synthase gene (AOS, LOC_Os03g12500) can be highly upregulated following brown planthopper (Nilaparvata lugens Stål, BPH) infestation. Then, we amplified the promoter of OsAOS1 and the β‐ glucuronidase reporter gene was used to analyze the expression pattern of the promoter. Through a series of 5ʹ truncated assays, three positive regulatory regions in response to BPH infestation in the promoter were identified. The transgenic plants, P1R123‐min 35S and P1TR1‐min 35S promoter‐driven snowdrop lectin (Galanthus nivalis agglutinin, GNA) gene, demonstrated the highest expression levels of GNA and lowest BPH survival. Our work identified a BPH‐inducible promoter and three positive regions within it. Transgenic rice with GNA driven by OsAOS1 promoter and positive regions exhibited an expected lethal effect on BPH. This study proved the application potential of BPH‐inducible promoter and provided a novel path for the selection of insect‐resistant tools in the future.
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