Skip to main content

Effect of nitrogen fertilizer on the resistance of rice near-isogenic lines with BPH resistance genes

Abstract

Background

Nitrogen is an essential macronutrient for plant growth and development. Crops with a high nitrogen input usually have high yields. However, outbreaks of brown planthoppers (Nilaparvata lugens; BPH) frequently occur on rice farms with excessive nitrogen inputs. Rice plants carrying BPH resistance genes are used for integrated pest management. Thus, the impact of nitrogen on the resistance of rice near-isogenic lines (NILs) with BPH resistance genes was investigated.

Results

We tested these NILs using a standard seedbox screening test and a modified bulk seedling test under different nitrogen treatments. The amount of nitrogen applied had an impact on the resistance of some lines with BPH resistance genes. In addition, three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) were further examined for antibiosis and antixenosis under varying nitrogen regimes. The N. lugens nymph population growth rate, honeydew excretion, female fecundity, and nymph survival rate on the three NILs were not affected by different nitrogen treatments except the nymph survival rate on NIL-BPH9 and the nymph population growth rate on NIL-BPH17. Furthermore, in the settlement preference test, the preference of N. lugens nymphs for IR24 over NIL-BPH9 or NIL-BPH17 increased under the high-nitrogen regime, whereas the preference of N. lugens nymphs for IR24 over NIL-BPH32 was not affected by the nitrogen treatments.

Conclusions

Our results indicated that the resistance of three tested NILs did not respond to different nitrogen regimes and that NIL-BPH17 exerted the most substantial inhibitory effect on N. lugens growth and development.

Background

Host plant resistance is a valuable resource for integrated pest management (IPM). Plants with resistance reduce not only herbivore damage but also pesticide usage. Antibiosis, antixenosis, and tolerance are the three categories of host plant resistance (Painter 1951; Smith 2005). Plants with antibiosis traits affect insect survival, whereas plants with antixenosis may influence insect behavior (Smith 2005). Plant tolerance is a unique trait in which a plant can withstand herbivore damage but does not affect insect growth and behavior (Smith 2005). Currently, insect-resistant varieties of major crops (rice, wheat, etc.) are widely used in IPM programs (Cohen et al. 1997; Jlibene and Nsarellah 2011; Nsarellah et al. 2003; Peñalver Cruz et al. 2011).

The brown planthopper (BPH), Nilaparvata lugens (Stål), is the major rice pest threatening rice production. N. lugens causes plant mortality symptom “hopper burn” and transmits plant viruses, such as grassy and ragged stunt viruses. Thirty nine BPH resistance genes in rice have been identified (Zhang et al. 2020). Twenty of them have been found in rice cultivars, whereas some have been identified in wild rice species, such as Oryza australiensis, O. officinalis, O. minuta, O. rufipogon, O. glaberrima, and O. nivara (Du et al. 2020). Furthermore, 14 BPH genes located on chromosomes 3, 4, 6, and 12 have been cloned and characterized (Cheng et al. 2013; Du et al. 2009, 2020; Guo et al. 2018; Ji et al. 2016; Liu et al. 2015; Ren et al. 2016; Tamura et al. 2014; Wang et al. 2015; Zhao et al. 2016). For example, BPH9 was found in the rice variety Pokkali and encodes a coiled-coil, nucleotide-binding, nucleotide-binding, and leucine-rich repeat domain (CC-NB-NB-LRR) protein (Zhao et al. 2016). BPH17 has been found in the rice cultivar Rathu Heenati and identified as a cluster of lectin receptor kinases (Liu et al. 2015). BPH32 was identified in PTB33 and contains a short consensus repeat (SCR) domain (Ren et al. 2016).

In our previous study, twelve near-isogenic lines (NILs) carrying one or two BPH resistance gene(s) were evaluated for resistance to environmental changes (high air temperature and high carbon dioxide concentration) (Kuang et al. 2021). Two of nine NILs with a single BPH resistance gene (BPH17 and BPH20) and two of three NILs pyramided with two BPH resistance genes (BPH9 + 32 and BPH18 + 32) maintained resistance against N. lugens under environmental changes (Kuang et al. 2021). Furthermore, NIL-BPH17 exerted a strong inhibitory effect on N. lugens growth and development despite the environmental changes. In addition, plants with the BPH17 resistance gene show resistance against the white-back planthopper [Sogatella furcifera (Horváth)] (Liu et al. 2015). BPH resistance genes are currently used in breeding programs for insect-resistant rice (Du et al. 2020; Jena et al. 2017; Nguyen et al. 2019; Xiao et al. 2016).

Since insect herbivores mainly obtain nutrients from host plants, the resource availability of the host plant is the main factor affecting insect herbivore growth and development (Awmack and Leather 2002). Nitrogen is an essential macronutrient for plant growth and development. Generally, crops with high nitrogen input have high production. However, nitrogen is also the limiting nutrient for insect herbivores. Insect herbivores feeding N-enriched host plants show enhanced fitness (Lu and Heong 2009; Lu et al. 2004; Prestidge 1982; Wier and Boethel 1995). For example, rice water weevils (Lissorhoptrus oryzophilus) feeding on high-nitrogen-treated plants showed increased adult feeding and oviposition preferences (Jiang and Cheng 2003). Furthermore, to obtain sufficient nitrogen, the midgut of Lepidoptera can digest large amounts of plant proteins, including Rubisco (Bhardwaj et al. 2014).

N. lugens outbreaks frequently occur on rice farms with excessive nitrogen input (Visarto et al. 2001). By increasing the host plant’s nitrogen content, insects may obtain sufficient nutrients to overcome plant resistance. Thus, we aimed to determine whether N. lugens feeding on rice plants with BPH resistance genes under high nitrogen input would overcome resistance. Therefore, we used twelve NILs with BPH resistance genes developed by the International Rice Research Institute (IRRI) to evaluate the impacts of nitrogen on resistance (Jena et al. 2017). These NILs were assessed by the standard seedbox screening test (SSST) and modified bulk seeding test (MBST). Furthermore, three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) were tested for antibiosis and antixenosis under different nitrogen treatments. Such information would provide evidence of the impact of nitrogen on BPH resistance genes and further reveal candidate BPH resistance genes for IPM programs.

Materials and methods

Plant materials

Taichung Native 1 (TN1), IR24, and twelve NILs with one or two BPH resistance genes were used in this study. Twelve NILs were initially obtained from the IRRI (Jena et al. 2017). IR24, the recurrent parent of the NILs, was obtained from the National Plant Genetic Resources Center, Taiwan Agricultural Research Institute, Taiwan (TARI). The susceptible control TN1 used for the SSST was obtained from Dr. Shu-Jen Wang, National Taiwan University. Seeds were sterilized with 2% NaOCl (CLOROX, California, United States) for 30 min in a shaker and further washed with distilled water for 10 min. The seeds were germinated on a moistened paper towel under dark conditions at 37 °C for 2 days.

Environmental setting

In this study, all plants were fertilized with ammonium sulfate, single superphosphate, and potassium chloride (Taiwan Fertilizer Company, Taiwan) to supply nitrogen, phosphate, and potassium, respectively. For nitrogen application, equivalent amounts of nitrogen were added to reach 0 kg ha− 1 (denoted N0), 50 kg ha− 1 (denoted N50), 100 kg ha− 1 (denoted N100), and 200 kg ha− 1 (denoted N200). The amounts of phosphate and potassium added were equivalent to 50 kg ha− 1 and 60 kg ha− 1, respectively. Before planting, basal fertilizer was applied at 30% for nitrogen, 100% for phosphate, and 40% for potassium. Plants and N. lugens were grown in a walk-in chamber with a day/night temperature of 30 °C/25 °C and a 12-h light/12-h dark cycle.

Insects

An N. lugens colony (biotype 1) was obtained from the Chiayi Agricultural Experimental Station, TARI. N. lugens was mass-reared on TN1 seedlings in an insect cage (BugBorm-4, Megaview, Taichung, Taiwan) in a walk-in chamber with a day/night temperature of 30 °C/25 °C and a 12-h light/ 12-h dark cycle (L/D).

SPAD value

The leaf chlorophyll contents of the three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) and IR24 were measured by a Soil Plant Analysis Development chlorophyll meter (SPAD 502 Plus Chlorophyll Meter 2900P, Konica Minolta, Osaka, Japan).

The SPAD is the alternative approach to measure the chlorophyll content without damaging leaf tissues. The average readings from the tip, middle, and base of the youngest expanded leaf of a 30-day-old rice plant were calculated. Each treatment included five replicate plants, and the experiment was repeated three times.

Standard seedbox screening test (SSST) and modified bulk seeding test (MBST)

Twelve NILs, IR24, and the susceptible control TN1 treated with different nitrogen applications (N0, N50, and N200) were evaluated for insect resistance by the SSST and MBST. Briefly, 24 seeds of each tested NIL/variety were sown in a row, and 20 seedlings were selected for the test. Fourteen days after sowing, 2nd- to 3rd-instar N. lugens were applied to the seedlings (8–10 N. lugens per seedling). For the SSST, the damage level was measured according to the standard evaluation system (IRRI 2013) when the susceptible control TN1 was dead. For the MBST, the seedling survival evaluation scale followed Jena et al. (2006). These experiments were repeated three times.

Population growth rate (PGR), honeydew excretion, fecundity, egg hatchability, survival rate, and settlement preference of N. lugens

Quantification of the PGR, honeydew excretion, fecundity, egg hatchability, survival rate, and settlement preference was performed using methods previously described by Kuang et al. (2021). Briefly, germinated seeds of three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) and IR24 were transferred to 150 ml glass beakers containing Kimura B solution (Yoshida et al. 1971). After seven days, the seedlings were transferred into plastic pots (one plant per pot) with paddy soil treated with basal fertilizer application. Three nitrogen applications (N50, N100, and N200) were applied in these experiments. At 30 days after germination, all branches except the main tiller were removed. The sample size (n) and the number of replicates (N) in most of the assays were N = 3 and n = 5, respectively; however, in the honeydew excretion assay, these values were N = 4 and n = 5, and in the settlement preference test, these values were n = 500.

Statistical analysis

All the data were analyzed using R software (v 4.0.5) (Team 2013). The SSST and MBST results were analyzed by two-way ANOVA, and the PGR, honeydew excretion, fecundity, egg hatchability, and survival rate data were analyzed by one-way ANOVA. The least significant difference test was used to detect differences at p < 0.05. In the multiple comparison procedure, Bonferroni’s correction method was applied to control the familywise error rate (FWER) to ensure a lower value than the nominal level of 0.05. The settlement preference data were analyzed using the standard z-test to evaluate whether or not N. lugens nymphs had settlement preference. Specifically, if they had no preference and selected the plants randomly, then the proportions of choosing IR24 and NIL-BPH9/17/32 would be equal to 0.5.

Results

SSST with different nitrogen treatments

Twelve NILs and their recurrent parent IR24 were evaluated for the impact of different nitrogen treatments on resistance against N. lugens using an SSST. The damage scores of the experimental plants were affected by the variety and treatment x variety interaction (p value < 0.001; Table 1). Under the no-nitrogen regime (N0), 9 NILs (NIL-BPH4, NIL-BPH9, NIL-BPH17, NIL-BPH20, NIL-BPH26, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a lower damage score than IR24, whereas 3 NILs (NIL-BPH10, NIL-BPH18, and NIL-BPH21) had similar scores to IR24 (Table 2). Under the low-nitrogen regime (N50), 9 NILs (NIL-BPH4, NIL-BPH9, NIL-BPH17, NIL-BPH18, NIL-BPH20, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a lower damage score than IR24, whereas 3 NILs (NIL-BPH10, NIL-BPH21, and NIL-BPH26) had similar scores to IR24 (Table 2). Under the high-nitrogen regime (N200), 8 NILs (NIL-BPH9, NIL-BPH17, NIL-BPH20, NIL-BPH21, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a lower damage score than IR24, whereas 3 NILs (NIL-BPH4, NIL-BPH10, and NIL-BPH18) had similar scores to IR24 (Table 2). In addition, NIL-BPH26 had a higher damage score than IR24 under N200. Compared with the N0 and N200 regimes, one NIL (NIL-BPH21) and IR24 showed variation in their resistance levels (Table 2). IR24 and NIL-BPH21 showed no resistance under N0 and N50 but gained resistance under N200. Overall, 4 NILs carrying a single BPH resistance gene (NIL-BPH9, NIL-BPH17, NIL-BPH20, and NIL-BPH32) and 3 NILs with gene pyramiding (NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) maintained their resistance under different nitrogen treatments.

Table 1 Two-way ANOVA of the SSST results of NIL responses to factors
Table 2 SSST of NILs under different nitrogen regimes

MBST with different nitrogen treatments

The resistance score of the tested plants was affected by the variety and treatment x variety interaction (p value < 0.001; Table 3). Under the no-nitrogen treatment (N0), 7 NILs (NIL-BPH9, NIL-BPH17, NIL-BPH20, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a higher resistance score than IR24, whereas 5 NILs (NIL-BPH4, NIL-BPH10, NIL-BPH18, NIL-BPH21, and NIL-BPH26) had a lower survival rate similar to IR24 (Table 4). Under the low-nitrogen treatment (N50), 7 NILs (NIL-BPH4, NIL-BPH9, NIL-BPH17, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a higher resistance score than IR24, and 5 NILs (NIL-BPH10, NIL-BPH18, NIL-BPH20, NIL-BPH21, and NIL-BPH26) had a lower survival rate similar to IR24 (Table 4). Under the high-nitrogen treatment (N200), 8 NILs (NIL-BPH4, NIL-BPH17, NIL-BPH20, NIL-BPH21, NIL-BPH32, NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) had a higher resistance score than IR24, whereas 4 NILs (NIL-BPH9, NIL-BPH10, NIL-BPH18, and NIL-BPH26) had a lower survival rate similar to IR24 (Table 4). Compared with the N0 and N200 regimes, two NILs (NIL-BPH4 and NIL-BPH21) showed variation in their resistance levels (Table 4). NIL-BPH4 and NIL-BPH21 showed no resistance under the N0 and N50 regimes but gained resistance under the N200 regime. Overall, 2 NILs carrying a single BPH resistance gene (NIL-BPH17 and NIL-BPH32) and 3 NILs with gene pyramiding (NIL-BPH2 + 32, NIL-BPH9 + 32, and NIL-BPH18 + 32) maintained their resistance under nitrogen treatments.

Table 3 Two-way ANOVA of the MBST results of NIL responses to factors
Table 4 MBST of NILs under different nitrogen regimes

Resistance of NIL-BPH9, NIL-BPH17, and NIL-BPH32 under nitrogen treatments

Based on the above data, three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) were selected to test for antibiosis and antixenosis effects under different nitrogen applications. The PGR, honeydew excretion, fecundity, egg hatchability, and survival rate were used to test for antibiosis effects, while a settlement preference test was used to test for an antixenosis effect. Nitrogen is the main factor affecting crop yield. No nitrogen application (N0) is not applicable in farming practices. Thus, three nitrogen treatments (N50, N100, and N200) were selected to study the effects further. The chlorophyll content of NIL-BPH9, NIL-BPH17, and NIL-BPH32 under the nitrogen treatments was measured. The SPAD value was not different among the tested plants with the same nitrogen treatment (Fig. 1). However, all tested varieties had higher SPAD values under the N200 treatment than under the other two nitrogen regimes (N50 and N100), except NIL-BPH17 and NIL-BPH32 under the N100 regime (Fig. 1).

Fig. 1
figure 1

The SPAD values of IR24, NIL-BPH9, NIL-BPH17, and NIL-BPH32 under different nitrogen regimes (N50, N100, and N200). Means in each column followed by the same capital letter do not differ significantly among varieties (p < 0.05). Means in each column followed by the same lowercase letter do not differ significantly among different nitrogen treatments. The error bars indicate the S.Es

A honeydew excretion assay was implemented as an indirect method to examine the phloem and xylem sap consumption of N. lugens. For phloem-derived honeydew, N. lugens feeding on NIL-BPH17 had lower phloem sap consumption than N. lugens feeding on IR24 under all nitrogen treatments, while N. lugens feeding on NIL-BPH9 and NIL-BPH32 had lower phloem sap consumption than N. lugens feeding on IR24 under the N200 treatment (Fig. 2a). Among the nitrogen treatments, no difference was found among N. lugens feeding on NIL-BPH9, NIL-BPH17, and NIL-BPH32 (Fig. 2a). For xylem-derived honeydew, N. lugens feeding on NIL-BPH9 and NIL-BPH17 had a lower xylem sap consumption than N. lugens feeding on IR24 under the N100 treatment (Fig. 2b). N. lugens feeding on IR24 under the N100 treatment had higher xylem sap consumption than that under the N50 treatment, whereas no difference was found among N. lugens feeding on NIL-BPH9, NIL-BPH17, and NIL-BPH32 (Fig. 2b).

Fig. 2
figure 2

Areas of honeydew excretion of N. lugens females feeding on IR24, NIL-BPH9, NIL-BPH17, and NIL-BPH32 under different nitrogen regimes. a Phloem-derived excretion. b Xylem-derived excretion. Means in each column followed by the same capital letter do not differ significantly among varieties (p < 0.05). Means in each column followed by the same lowercase letter do not differ significantly among different nitrogen treatments. The error bars indicate the S.Es

The PGR was used as the growth parameter of N. lugens. There was no difference between IR24 and NIL-BPH9 or NIL-BPH32 under any nitrogen treatments (Fig. 3). However, N. lugens had a lower PGR on NIL-BPH17 than on IR24 under the N50 and N200 treatments but similar PGRs on NIL-BPH17 and IR24 under the N100 treatment (Fig. 3). In addition, N. lugens feeding on NIL-BPH17 had a lower PGR under the N50 and treatment than under the N100 treatment, whereas no difference was found among IR24, NIL-BPH9, and NIL-BPH32 under any treatment (Fig. 3). The nymph survival rate of N. lugens on these NILs under different nitrogen treatments was further examined. N. lugens nymphs on NIL-BPH17 had a lower survival rate than those on IR24 under the N200 treatment, whereas no difference was found among IR24, NIL-BPH9, and NIL-BPH32 under any of the treatments (Fig. 4). Within the same variety, N. lugens nymphs feeding on IR24 under the N200 regime had a higher survival rate than those feeding on IR24 under low nitrogen application (N50 and N100 treatments), whereas there was no difference with NIL-BPH17 and NIL-BPH32 (Fig. 4).

Fig. 3
figure 3

Population growth rate of N. lugens nymphs feeding on IR24, NIL-BPH9, NIL-BPH17, and NIL-BPH32 under different nitrogen regimes. Means in each column followed by the same capital letter do not differ significantly among varieties (p < 0.05). Means in each column followed by the same lowercase letter do not differ significantly among different nitrogen treatments. The error bars indicate the S.Es

Fig. 4
figure 4

Nymph survival rate of N. lugens feeding on IR24, NIL-BPH9, NIL-BPH17, and NIL-BPH32 under different nitrogen regimes. Means in each column followed by the same capital letter do not differ significantly among varieties (p < 0.05). Means in each column followed by the same lowercase letter do not differ significantly among different nitrogen treatments. The error bars indicate the S.Es

Using a no-choice assay, female fecundity under different nitrogen applications was examined. N. lugens female adults on NIL-BPH32 had lower fecundity than those on IR24 under the N100 treatment, whereas N. lugens female adults on NIL-BPH17 had lower fecundity than those on IR24 under all nitrogen treatments (Fig. 5). In the settlement preference test, N. lugens nymphs preferred settling on IR24 over NIL-BPH9 from 48 to 120 h under the N50 treatment (Fig. 6a). Under the N100 and N200 treatments, N. lugens nymphs preferred IR24 from 3 to 120 h, except at the 48-h time point under the N200 treatment (Fig. 6b, c). When comparing IR24 and NIL-BPH17, N. lugens nymphs preferred IR24 at 120 h under the N50 treatment and preferred IR24 at 24 h under the N100 and N200 treatments (Fig. 7). For comparing IR24 and NIL-BPH32, N. lugens nymphs preferred IR24 from 3 to 120 h under all nitrogen treatments except the 3 and 120 h time points under the N100 treatment (Fig. 8).

Fig. 5
figure 5

 N. lugens female fecundity on IR24, NIL-BPH9, NIL-BPH17, and NIL-BPH32 under different nitrogen regimes. Means in each column followed by the same capital letter do not differ significantly among varieties (p < 0.05). Means in each column followed by the same lowercase letter do not differ significantly among different nitrogen treatments. The error bars indicate the S.Es

Fig. 6
figure 6

Choice test of N. lugens nymphs on IR24 and NIL-BPH9 under different nitrogen regimes. a N50. b N100. c N200. The asterisks indicate differences between IR24 and NIL-BPH9 as *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance

Fig. 7
figure 7

Choice test of N. lugens nymphs on IR24 and NIL-BPH17 under different nitrogen regimes. a N50. b N100. c N200. The asterisks indicate differences between IR24 and NIL-BPH17 as *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance

Fig. 8
figure 8

Choice test of N. lugens nymphs on IR24 and NIL-BPH32 under different nitrogen regimes. a N50. b N100. c N200. The asterisks indicate differences between IR24 and NIL-BPH32 as *p < 0.05; **p < 0.01; ***p < 0.001; ns no significance

Discussion

Plants with insect resistance traits are keys to IPM programs. Since BPH-resistant rice varieties have been used in the market, the environmental impact on their resistance has been noticed (Horgan et al. 2021; Kuang et al. 2021). Nitrogen input is highly correlated with crop yield. However, excessive nitrogen input may not increase crop production but instead benefit insect pests. Therefore, the amount of nitrogen input should be appropriate and considered in the IPM program. In this study, we examined twelve NILs with BPH resistance genes under different nitrogen regimes. Three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) maintained a low damage score under varying nitrogen applications. High nitrogen input would increase the SPAD value, the indicator of chlorophyll content, in the leaf tissues. Based on the N. lugens growth parameters, the resistance of the three tested NILs did not respond to different nitrogen regimes, whereas NIL-BPH17 exerted the strongest inhibitory effect on N. lugens growth and development.

The fitness of N. lugens increases with increases in the plant nitrogen content in rice (Lu et al. 2004; Rashid et al. 2016, 2017a, b). Increasing the soil nitrogen level increases the survival rate and weight of N. lugens nymphs and shortens their developmental period (Horgan et al. 2016, 2018; Rashid et al. 2017b). Some of these results were consistent with those of our study (Fig. 4). In addition, the biomass of N. lugens nymphs on a susceptible variety (T65) increased under a high-nitrogen regime (Srinivasan et al. 2015). The impact of nitrogen on the fitness of N. lugens adults is not conclusive. The adult longevity, fecundity, hatchability, weight, and survival rate of N. lugens increases with increases in the nitrogen content of the host plants (Rashid et al. 2016, 2017b). However, Horgan et al. (2016) reported that nitrogen fertilizer treatments did not affect fecundity or egg mortality. Our study supports the last finding. In addition, under a high-nitrogen regime, N. lugens fecundity increased across generations (Lu et al. 2004). This study revealed that N. lugens outbreaks may frequently occur in nitrogen-enriched crops (Lu et al. 2004; Horgan et al. 2021) reported that increasing nitrogen input would reduce resistance in rice but enhance its tolerance to N. lugens. Our study yielded similar results on IR24 (Table 2). However, in our study, N. lugens feeding on three tested NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) under high nitrogen input did not overcome the resistance. These results indicated that these three BPH resistance genes would benefit rice breeding programs.

Breeding insect-resistant varieties with insect resistance genes is an effective and environmentally friendly strategy for IPM programs. Several technologies, including marker-assisted selection and gene editing, accelerate the breeding process. BPH resistance genes have been developed in several rice varieties, such as 9311, IR24, and T65 (Jena et al. 2017; Nguyen et al. 2019; Xiao et al. 2016). Furthermore, NILs with resistance to other phloem feeders, including the white-backed planthopper (S. furcifera), gall midge (Orseolia oryzae), and green rice leafhopper (N. cincticeps), have also been developed (Fujita et al. 2010; Himabindu et al. 2010; Yamasaki et al. 2003). However, because N. lugens has multiple biotypes and is prone to adaptation, rice varieties with a single resistance gene may show a reduction in resistance within a few years (Jena and Kim 2010). Furthermore, several BPH resistance genes lose their efficacy under environmental changes (Kuang et al. 2021). Thus, pyramiding multiple genes would be a better strategy. It has been reported that pyramided genes have a synergistic effect (Hu et al. 2013; Jena et al. 2017; Qiu et al. 2012). In our study, under N0 treatment, NIL-BPH9 + 32 and NIL-BPH18 + 32 had lower damage scores in the SSST and higher resistance in the MBST than the NILs with a single resistance gene (BPH9, BPH18, and BPH32). Furthermore, NIL-BPH9 + 32 and NIL-BPH18 + 32 showed a similar trend under environmental change (Kuang et al. 2021). Thus, pyramiding genes in one variety not only prevents the loss of efficacy but also enhances resistance to environmental changes, including climate change and varying nitrogen inputs.

Climate change impact and excessive nitrogen input are the two critical challenges to our crop production. Therefore, we would like to use this unique NIL set to find out BPH genes that would maintain the resistance under stress. Based on the findings of this study and our previous results, NIL-BPH17 maintained resistance against N. lugens under not only environmental changes (high atmospheric temperature and high CO2 concentration) but also varying nitrogen applications (Kuang et al. 2021). Furthermore, our results showed that increasing the nitrogen level enhanced the preferences of N. lugens for IR24 from 120 h to 24 h after the experiment (Fig. 7). In addition, with environmental changes, the preferences of N. lugens nymphs for IR24 and NIL-BPH17 accelerated from 24 h to 6 h after the experiment (Kuang et al. 2021). Thus, BPH17 may be the best BPH resistance gene for insect resistance breeding programs in rice.

Conclusions

The impact of nitrogen on the resistance of twelve NILs with BPH resistance genes against N. lugens was examined. Nitrogen input affected some of the tested lines with BPH resistance genes. However, three NILs (NIL-BPH9, NIL-BPH17, and NIL-BPH32) did not show changes in resistance with different nitrogen regimes, while NIL-BPH17 had the strongest inhibitory effect on N. lugens growth and development. These results provide valuable information for IPM programs.

Availability of data and materials

The data used and analyzed for the current study can be obtained from the corresponding author.

References

  • Awmack CS, Leather SR (2002) Host plant quality and fecundity in herbivorous insects. Ann Rev Entomol 47:817–844

    CAS  Article  Google Scholar 

  • Bhardwaj U, Bhardwaj A, Kumar R, Leelavathi S, Reddy VS, Mazumdar-Leighton S (2014) Revisiting Rubisco as a protein substrate for insect midgut proteases. Arch Insect Biochem Physiol 85:13–35

    CAS  PubMed  Article  Google Scholar 

  • Cheng X, Wu Y, Guo J, Du B, Chen R, Zhu L, He G (2013) A rice lectin receptor-like kinase that is involved in innate immune responses also contributes to seed germination. Plant J 76:687–698

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Cohen MB, Alam SN, Medina EB, Bernal CC (1997) Brown planthopper, Nilaparvata lugens, resistance in rice cultivar IR64: mechanism and role in successful N. lugens management in Central Luzon, Philippines. Entomol Exp Appl 85:221–229

    Article  Google Scholar 

  • Du B, Zhang W, Liu B, Hu J, Wei Z, Shi Z, He R, Zhu L, Chen R, Han B, He G (2009) Identification and characterization of Bph14, a gene conferring resistance to brown planthopper in rice. Proc Natl Acad Sci USA 106:22163–22168

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Du B, Chen R, Guo J, He G (2020) Current understanding of the genomic, genetic, and molecular control of insect resistance in rice. Mol Breeding 40:24

    CAS  Article  Google Scholar 

  • Fujita D, Yoshimura A, Yasui H (2010) Development of near-isogenic lines and pyramided lines carrying resistance genes to green rice leafhopper (Nephotettix cincticeps Uhler) with the Taichung 65 genetic background in rice (Oryza sativa L.). Breed Sci 60:18–27

    CAS  Article  Google Scholar 

  • Guo J, Xu C, Wu D, Zhao Y, Qiu Y, Wang X, Ouyang Y, Cai B, Liu X, Jing S (2018) Bph6 encodes an exocyst-localized protein and confers broad resistance to planthoppers in rice. Nat Genet 50:297–306

    CAS  PubMed  Article  Google Scholar 

  • Himabindu K, Suneetha K, Sama V, Bentur J (2010) A new rice gall midge resistance gene in the breeding line CR57-MR1523, mapping with flanking markers and development of NILs. Euphytica 174:179–187

    CAS  Article  Google Scholar 

  • Horgan FG, Srinivasan TS, Naik BS, Ramal AF, Bernal CC, Almazan MLP (2016) Effects of nitrogen on egg-laying inhibition and ovicidal response in planthopper-resistant rice varieties. Crop Prot 89:223–230

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Horgan FG, Cruz AP, Bernal CC, Ramal AF, Almazan MLP, Wilby A (2018) Resistance and tolerance to the brown planthopper, Nilaparvata lugens (Stål), in rice infested at different growth stages across a gradient of nitrogen applications. Field Crops Res 217:53–65

    PubMed  PubMed Central  Article  Google Scholar 

  • Horgan FG, de Freitas TFS, Crisol-Martínez E, Mundaca EA, Bernal CC (2021) Nitrogenous fertilizer reduces resistance but enhances tolerance to the brown planthopper in fast-growing, moderately resistant rice. Insects 12:989

    PubMed  PubMed Central  Article  Google Scholar 

  • Hu J, Cheng M, Gao G, Zhang Q, Xiao J, He Y (2013) Pyramiding and evaluation of three dominant brown planthopper resistance genes in the elite indica rice 9311 and its hybrids. Pest Manag Sci 69:802–808

    CAS  PubMed  Article  Google Scholar 

  • IRRI (2013) Standard evaluation system for rice. International Rice Research Institute, Los Baños

    Google Scholar 

  • Jena KK, Kim SM (2010) Current status of brown planthopper (BPH) resistance and genetics. Rice 3:161–171

    Article  Google Scholar 

  • Jena KK, Jeung JU, Lee JH, Choi HC, Brar DS (2006) High-resolution mapping of a new brown planthopper (BPH) resistance gene, Bph18 (t), and marker-assisted selection for BPH resistance in rice (Oryza sativa L.). Theor Appl Genet 112:288–297

    CAS  PubMed  Article  Google Scholar 

  • Jena KK, Hechanova SL, Verdeprado H, Prahalada GD, Kim S-R (2017) Development of 25 near-isogenic lines (NILs) with ten BPH resistance genes in rice (Oryza sativa L.): Production, resistance spectrum, and molecular analysis. Theor Appl Genet 130:2345–2360

    CAS  PubMed  Article  Google Scholar 

  • Ji H, Kim SR, Kim YH, Suh JP, Park HM, Sreenivasulu N, Misra G, Kim SM, Hechanova SL, Kim H, Lee GS, Yoon UH, Kim TH, Lim H, Suh SC, Yang J, An G, Jena KK (2016) Map-based cloning and characterization of the BPH18 gene from wild rice conferring resistance to brown planthopper (BPH) insect pest. Sci Rep 6:34376

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Jiang M, Cheng J (2003) Feeding, oviposition and survival of overwintered rice water weevil (Coleoptera: Curculionidae) adults in response to nitrogen fertilization of rice at seedling stage. Appl Entomol Zool 38:543–549

    Article  Google Scholar 

  • Jlibene M, Nsarellah N (2011) Wheat breeding in Morocco, a historical perspective. The world wheat book: history of wheat breeding. Lavoisier, Paris

    Google Scholar 

  • Kuang YH, Fang YF, Lin SC, Tsai SF, Yang ZW, Li CP, Huang SH, Hechanova SL, Jena KK, Chuang WP (2021) The impact of climate change on the resistance of rice near-isogenic lines with resistance genes against brown planthopper. Rice 14:1–15

    Article  Google Scholar 

  • Liu Y, Wu H, Chen H, Liu Y, He J, Kang H, Sun Z, Pan G, Wang Q, Hu J, Hu J, Zhou F, Zhou K, Zheng X, Ren Y, Chen L, Wang Y, Zhao Z, Lin Q, Wu F, Zhang X, Guo X, Cheng X, Jiang L, Wu C, Wang H, Wan J (2015) A gene cluster encoding lectin receptor kinases confers broad-spectrum and durable insect resistance in rice. Nat Biotechnol 33:301–305

    CAS  PubMed  Article  Google Scholar 

  • Lu ZX, Heong KL (2009) Effects of nitrogen-enriched rice plants on ecological fitness of planthoppers. Planthoppers: new threats to the sustainability of intensive rice production systems in Asia. International Rice Research Institute, Los Baños

    Google Scholar 

  • Lu ZX, Heong KL, Yu XP, Hu C (2004) Effects of plant nitrogen on ecological fitness of the brown planthopper, Nilaparvata lugens Stal. in rice. J Asia Pac Entomol 7:97–104

    Article  Google Scholar 

  • Nguyen CD, Verdeprado H, Zita D, Sanada-Morimura S, Matsumura M, Virk PS, Brar DS, Horgan FG, Yasui H, Fujita D (2019) The development and characterization of near-isogenic and pyramided lines carrying resistance genes to brown planthopper with the genetic background of japonica rice (Oryza sativa L.). Plants 8:498

    CAS  PubMed Central  Article  Google Scholar 

  • Nsarellah N, Amri A, Nachit M, Bouhssini ME, Lhaloui S, Lorenzoni C (2003) New durum wheat with Hessian fly resistance from Triticum araraticum and T. carthlicum in Morocco. Plant Breeding 122:435–437

    Article  Google Scholar 

  • Painter RH (1951) Insect resistance in crop plants. The University Press of Kansas, Lawrence

    Book  Google Scholar 

  • Peñalver Cruz A, Arida A, Heong KL, Horgan FG (2011) Aspects of brown planthopper adaptation to resistant rice varieties with the Bph3 gene. Entomol Exp Appl 141:245–257

    Article  CAS  Google Scholar 

  • Prestidge R (1982) Instar duration, adult consumption, oviposition and nitrogen utilization efficiencies of leafhoppers feeding on different quality food (Auchenorrhyncha: Homoptera). Ecol Entomol 7:91–101

    Article  Google Scholar 

  • Qiu Y, Guo J, Jing S, Zhu L, He G (2012) Development and characterization of japonica rice lines carrying the brown planthopper-resistance genes BPH12 and BPH6. Theor Appl Genet 124:485–494

    CAS  PubMed  Article  Google Scholar 

  • Rashid M, Jahan M, Islam K (2016) Response of adult brown planthopper Nilaparvata lugens (Stål) to rice nutrient management. Neotrop Entomol 45:588–596

    CAS  PubMed  Article  Google Scholar 

  • Rashid M, Ahmed N, Jahan M, Islam K, Nansen C, Willers J, Ali M (2017a) Higher fertilizer inputs increase fitness traits of brown planthopper in rice. Sci Rep 7:1–16

    CAS  Article  Google Scholar 

  • Rashid M, Jahan M, Islam KS, Latif MA (2017) Ecological fitness of brown planthopper, Nilaparvata lugens (Stål), to rice nutrient management. Ecol Process 6:1–10

    Article  Google Scholar 

  • Ren J, Gao F, Wu X, Lu X, Zeng L, Lv J, Su X, Luo H, Ren G (2016) Bph32, a novel gene encoding an unknown SCR domain-containing protein, confers resistance against the brown planthopper in rice. Sci Rep 6:37645

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Smith CM (2005) Plant resistance to arthropods: molecular and conventional approaches. Springer Science & Business Media, Berlin

    Book  Google Scholar 

  • Srinivasan TS, Almazan MLP, Bernal CC, Fujita D, Ramal AF, Yasui H, Subbarayalu MK, Horgan FG (2015) Current utility of the BPH25 and BPH26 genes and possibilities for further resistance against plant-and leafhoppers from the donor cultivar ADR52. Appl Entomol Zool 50:533–543

    CAS  Article  Google Scholar 

  • Tamura Y, Hattori M, Yoshioka H, Yoshioka M, Takahashi A, Wu J, Sentoku N, Yasui H (2014) Map-based cloning and characterization of a brown planthopper resistance gene BPH26 from Oryza sativa L. ssp. indica cultivar ADR52. Sci Rep 4:5872

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  • Team RC (2013) R: a language and environment for statistical computing. Team RC, Austria

    Google Scholar 

  • Visarto P, Zalucki MP, Nesbitt HJ, Jahn GC (2001) Effect of fertilizer, pesticide treatment, and plant variety on the realized fecundity and survival rates of brown planthopper, Nilaparvata lugens (Stål) (Homoptera: Delphacidae)—generating outbreaks in Cambodia. J Asia Pac Entomol 4:75–84

    Article  Google Scholar 

  • Wang Y, Cao L, Zhang Y, Cao C, Liu F, Huang F, Qiu Y, Li R, Lou X (2015) Map-based cloning and characterization of BPH29, a B3 domain-containing recessive gene conferring brown planthopper resistance in rice. J Exp Bot 66:6035–6045

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  • Wier AT, Boethel DJ (1995) Feeding, growth, and survival of soybean looper (Lepidoptera: Noctuidae) in response to nitrogen fertilization of nonnodulating soybean. Environ Entomol 24:326–331

    Article  Google Scholar 

  • Xiao C, Hu J, Ao YT, Cheng MX, Gao GJ, Zhang QL, He GC, He YQ (2016) Development and evaluation of near-isogenic lines for brown planthopper resistance in rice cv. 9311. Sci Rep 6:1–10

    Article  CAS  Google Scholar 

  • Yamasaki M, Yoshimura A, Yasui H (2003) Genetic basis of ovicidal response to whitebacked planthopper (Sogatella furcifera Horváth) in rice (Oryza sativa L.). Mol Breeding 12:133–143

    CAS  Article  Google Scholar 

  • Yoshida S, Forno DA, Cock JH (1971) Laboratory manual for physiological studies of rice. International Rice Research Institute, Los Baños

    Google Scholar 

  • Zhang Y, Gang Q, Qianqian M, Minyi W, Xinghai Y, Zengfeng M, Haifu L, Chi L, Zhenjing L, Fang L (2020) Identification of major locus Bph35 resistance to brown planthopper in rice. Rice Sci 27:237–245

    Article  Google Scholar 

  • Zhao Y, Huang J, Wang Z, Jing S, Wang Y, Ouyang Y, Cai B, Xin XF, Liu X, Zhang C, Pan Y, Ma R, Li Q, Jiang W, Zeng Y, Shangguan X, Wang H, BoDu, Zhu L, Xu X, Feng Y-Q, He SY, Chen R, Zhang Q, He G (2016) Allelic diversity in an NLR gene BPH9 enables rice to combat planthopper variation. Proc Natl Acad Sci USA 113:12850–12855

    CAS  PubMed  PubMed Central  Article  Google Scholar 

Download references

Acknowledgements

The IR24 seeds were obtained from the National Plant Genetic Resources Center, Taiwan Agricultural Research Institute, Taiwan.

Funding

This work was supported by grants from the Ministry of Science and Technology of Taiwan (Grant Number 110-2313-B-002-026-MY3 to W-P. C.) and National Taiwan University (Taiwan; Grant Number NTU-109L7864 to W-P. C.).

Author information

Authors and Affiliations

Authors

Contributions

WC conceived the project; SL and WC designed the experiments; SL, YL, FH, CW, YK, CS, ZY, CL, SHH, CTL, SLH and KJ performed the experiments; SL, YL, and ST analyzed the data; and SL, ST, ZY, CPL, SHH, CTL, SLH, KJ, and WC wrote the manuscript. All authors provided feedback on the data interpretation. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Wen-Po Chuang.

Ethics declarations

Ethics approval and consent to participate

Not applicable.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Additional information

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Rights and permissions

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/.

Reprints and Permissions

About this article

Verify currency and authenticity via CrossMark

Cite this article

Lin, SC., Li, Y., Hu, FY. et al. Effect of nitrogen fertilizer on the resistance of rice near-isogenic lines with BPH resistance genes. Bot Stud 63, 16 (2022). https://doi.org/10.1186/s40529-022-00347-8

Download citation

  • Received:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1186/s40529-022-00347-8

Keywords

  • Nilaparvata lugens
  • Nitrogen
  • BPH9
  • BPH17
  • BPH32