Detection of Yeast-like Symbionts in Brown Planthopper Reared on Different Resistant Rice Varieties Combining DGGE and Absolute Quantitative Real-Time PCR

The brown planthopper (BPH), Nilaparvata lugens, is a serious pest of rice throughout Asia. Yeast-like symbionts (YLS) are endosymbionts closely linked with the development of BPH and the adapted mechanism of BPH virulence to resistant plants. In this study, we used semi-quantitative DGGE and absolute quantitative real-time PCR (qPCR) to quantify the number of the three YLS strains (Ascomycetes symbionts, Pichia-like symbionts, and Candida-like symbionts) that typically infect BPH in the nymphal stages and in newly emerged female adults. The quantities of each of the three YLS assessed increased in tandem with the developing nymphal instar stages, peaking at the fourth instar stage, and then declined significantly at the fifth instar stage. However, the amount of YLS present recovered sharply within the emerging adult females. Additionally, we estimated the quantities of YLS for up to eight generations after their inoculation onto resistant cultivars (Mudgo, ASD7, and RH) to reassociate the dynamics of YLS with the fitness of BPH. The minimum number of each YLS was detected in the second generation and gradually increased from the third generation with regard to resistant rice varieties. In addition, the Ascomycetes symbionts of YLS were found to be the most abundant of the three YLS strains tested for all of the development stages of BPH.

Repellency Mechanism of Natural Guar Gum-Based Film Incorporated with Citral against Brown Planthopper, Nilaparvata lugens (Stål) (Hemiptera: Delphacidae)

Using of plant essential oil that coevolved as a defense mechanism against agriculture insects is an alternative means of controlling many insect pests. In order to repel brown planthoppers (BPHs), the most notorious rice insect pest, a new film based on guar gum incorporated with citral (GC film) was formulated, which was effective while being environmentally friendly. In this paper, the effect and mechanism of GC film repellency against BPHs were determined. Repellent activity test and olfactory reaction analysis showed that GC film had repellency effect against BPHs, with repellency of 60.00% and 73.93%, respectively. The result of olfactory reaction indicated that GC film repellency against BPHs relied on smell. EPG analysis showed the proportion and mean duration of np waveform were significantly higher than in CK and increased following the treatment concentration, which indicated that GC film affected the recognition of BPHs to rice. Further analysis by RNA sequencing analysis showed a total of 679 genes were significantly upregulated and 284 genes were significantly downregulated in the BPHs fed on the rice sprayed with GC film compared to control. Odorant-binding protein (OBP) gene 797 and gustatory receptor gene (GR)/odorant receptor (OR) gene 13110 showed a significant decrease in differential expression and significant increase in differential expression, respectively. There were 0.66 and 2.55 differential expression multiples between treated BPHs and control, respectively. According to the results described above, we reasoned that GC film repellency against BPHs due to smell, by release of citral, caused the recognition difficulties for BPHs to rice, and OBP gene 797 and GR/OR gene 13110 appeared to be the crucial candidate genes for GC film repellency against BPHs. The present study depicted a clear and consistent repellency effect for GC film against BPHs and preliminarily clarified the mechanism of GC film as a repellent against BPHs, which might offer an alternative approach for control of BPHs in the near future. Our results could also help in the development and improvement of GC films.

Seasonal variation in population and biochemical contents of brown planthopper, Nilaparvata lugens (Stål)

Aim: The present study was undertaken on population dynamics and estimation of protein, water-soluble carbohydrates and glycogen contents in the brown planthopper, Nilaparvata lugens to explore their migratory behaviour. Methodology: Brown planthopper populations were monitored and collected using sweep nets from 23rd standard meteorological week (SMW) to 47th SMW during 2017 and 2018 rainy seasons to understand population dynamics of the pest. The protein, water-soluble carbohydrates and glycogen contents were estimated from the pest samples collected during 36th to 44th SMW in 2017 and 2018 rainy seasons. Results: Brown planthopper population were not observed in rice farm during 23rd SMW to 28th SMW during two years of study. However, macropterous form of the pest first appeared in the farm during 29th SMW and peaked during 43rd SMW. Thereafter, population declined and disappeared after 47th SMW during both the years. Water-soluble carbohydrates and glycogen contents varied significantly different weeks which remained low during 36th-39th SMW, however, increased gradually towards the end of the rainy season 2017 and 2018. On the other hand, protein content significantly varied among different weeks unlike the trend of water-soluble carbohydrates and glycogen. Interpretation: The study revealed the absence of brown plant hopper during summer season preceding rainy season, and the accumulation of bio-chemical compounds towards the end of rainy season under Delhi environment is perhaps suggestive of migration of the pest from unknown areas during rainy season to Delhi and likely preparedness of the pest for emigration to safer areas from Delhi, respectively.

Effects of Different Nutritional Conditions on the Growth and Reproduction of Nilaparvata lugens (Stål)

Growth and reproduction are the two most basic life processes of organisms and the distribution of energy in these processes is a core issue of the life history of organisms. Nilaparvata lugens (Stål), the brown planthopper (BPH), is a single-feeding rice pest. In the present study, this species was used as a model for testing the effects of nutritional conditions on various growth and reproduction indicators. First, the third-instar nymphs were fed with three different concentrations (100, 50, and 25%) of artificial diet until the second day of adulthood. The results showed that as the nutrient concentration decreased, the body development and oviposition of BPH were hindered. The total lipid content in the fat bodies was also significantly reduced. RT-PCR analysis showed compared to the 100% concentration group, the expression levels of vitellogenin (Vg) genes in the fifth-instar nymphs, adults, and in different tissues (ovary, fat body, and other tissues) were significantly decreased in the 50 and 25% treatment groups. Western blot analysis showed that Vg protein expression was highest in the 100% group, followed by the 50% group, with no expression in the 25% group. These results indicate that growth and reproduction in the BPH are regulated by, or correlated with, nutrient concentration. This study is of great significance as it reveals the adaptive strategies of the BPH to nutritional deficiencies and it also provides valuable information for the comprehensive control of this pest.

SSR MARKERS REVEALED GENETIC DIVERGENCE OF RICE BROWN PLANTHOPPER POPULATIONS MAINTAINED ON TWO SETS OF DIFFERENTIAL HOST VARIETIES

<p>Resistance screening of promising rice lines in Indonesia requires the use of brown planthopper (BPH) biotypes 1, 2, and 3. Three BPH populations have been raised as biotypes 1, 2, and 3 on differential rice host of improved varieties Pelita I-1 (no <em>Bph </em>gene), IR26 (<em>Bph1</em>), and IR42 (<em>bph2</em>), respectively. Three alternative populations have also been developed on the respective traditional varieties TN1 (no <em>Bph </em>gene), Mudgo (<em>Bph1</em>), and ASD7 (<em>bph2</em>). Although these populations displayed two virulent patterns other than biotype 1 to 3 phenotypes, they were expected to be discriminated into two virulence groups by SSR analysis. The study aimed to investigate the level of genetic variation among the six BPH populations using SSR markers and to relate it with the observed virulence patterns. Genotyping of 30 females with 29 polymorphic SSR markers revealed higher genetic parameter values in populations reared on improved varieties than those on traditional varieties. This difference was marked as two population clusters in PCoA plots corresponding to the host variety type, in contrast to the previous assumption that clustering would be based on virulence patterns. The presence of individuals with unwanted virulence allele, either resulting from contamination during the long period of rearing or lack of host adaptation period, is suspected. The result of this study indicates that the six populations are not suitable for resistance screening. Virulence selection must be performed until they attain biotype 1 to 3 phenotypes which can be genetically separated by DNA markers.</p>