Lipid composition differs in diapause and nondiapause states of spotted stem borer, Chilo partellus.

Spotted stem borer, Chilo partellus, undergoes larval diapause (hibernation and aestivation), and depends on the food reserve accumulated during feeding stage for its survival. Lipids are the primary source of energy during diapause, and essential for different cellular, biochemical and physiological functions. However, there is no information on lipid and lipophilic compound contents during different stages of hibernation, aestivation and nondiapause in C. partellus. Thus, we compared the concentration and composition of lipids in pre-diapause, diapause and post-diapause stages of hibernation and aestivation with nondiapause stages of C. partellus. The studies revealed significant differences in total lipids and various lipophilic compounds during different stages of diapause as compared to nondiapause C. partellus. The total lipids were significantly lower during diapause stage of aestivation and hibernation as compared to nondiapause larvae. Further, the linoleic acid, Methyl 3-methoxytetradecanoate, and l-(+)-Ascorbic acid 2,6-dihexadecanoate were significantly lower, and oleic and palmitoleic acids greater during pre-diapause and diapause stages of hibernation and aestivation as compared to nondiapause larvae. The cholesterol content was significantly greater during pre-diapause stage of hibernation, and diapause and post-diapause stages of aestivation as compared to nondiapause stages. The unsaturation ratio was significantly higher in the pre-diapause and diapause stages and lower in post-diapause stage of aestivation than the hibernation and nondiapause states. This study provides insights on differential lipid profiles during different phases of diapause, which could be useful for further understanding biochemical and physiological cross-talk, and develop target-specific technologies for the management of C. partellus.

Diapause-induced shift in the content of major carbohydrates in Chilo partellus (Swinhoe).

Although several aspects like diapause determining factors, population structure, reproductive physiology, and genetics of diapause have been investigated, there is no clarity on carbohydrate energetics during larval diapause in Chilo partellus (Swinhoe). Present studies revealed significant variation between the nondiapausing and diapausing C. partellus for total carbohydrates, glycogen, sorbitol, and trehalose contents in different body parts, life stages, and for body parts × life stages interaction. Total carbohydrate content started declining, while sorbitol and trehalose increased in all the body parts as the C. partellus larvae progressed from prediapausing to diapausing state. However, glycogen content spiked in all the body parts at prediapausing stage, which then declined during diapause. Among the body parts, total carbohydrate content was significantly greater in the hemolymph as compared to other body parts of both larvae and pupae of C. partellus. Glycogen content was significantly greater in the larval fat bodies and pupal hemolymph as compared to their other body parts. In diapausing larvae, sorbitol and trehalose were greater in the integument than in other body parts. Furthermore, there was spike in trehalose and decrease in sorbitol in all the body parts of pupae from diapausing than those from nondiapausing larvae. These findings suggest that the diapause alterate and/or fluctuate major carbohydrates in different body parts of both larvae and pupae of C. partellus. This information will be helpful in better understanding the diapause energetics and overwintering metabolic cryoprotection in insects.

Whole genome sequencing of spotted stem borer, Chilo partellus, reveals multiple genes encoding enzymes for detoxification of insecticides.

Spotted stem borer, Chilo partellus, is the most important constraint for increasing the production and productivity of maize and sorghum, the two major coarse cereals in Asia and Africa. The levels of resistance to this pest in the cultivated germplasm are low to moderate, and hence, farmers have to use insecticides for effective control of this pest. However, there is no information on the detoxification mechanisms in C. partellus, which is one of the constraints for deployment of appropriate insecticides to control this pest. The ability to detoxify insecticides varies across insect populations, and hence, we sequenced different populations of C. partellus to identify and understand detoxification mechanisms to devise appropriate strategies for deployment of different insecticides for controlling this pest. Larval samples were sequenced from three different cohorts of C. partellus using the Illumina HiSeq 2500 platform. The data were subjected to identify putative genes that are involved in detoxification on insecticides in our cohort insect species. These studies resulted in identification of 64 cytochrome P450 genes (CYP450s), and 36 glutathione S-transferases genes (GSTs) encoding metabolic detoxification enzymes, primarily responsible for xenobiotic metabolism in insects. A total of 183 circadian genes with > 80% homolog and 11 olfactory receptor genes that mediate chemical cues were found in the C. partellus genome. Also, target receptors related to insecticide action, 4 acetylcholinesterase (AChE), 14 γ-aminobutyric acid (GABA), and 15 nicotinic acetylcholine (nAChR) receptors were detected. This is the first report of whole genome sequencing of C. partellus useful for understanding mode of action of different insecticides, and mechanisms of detoxification and designing target-specific insecticides to develop appropriate strategies to control C. partellus for sustainable crop production.

The amino acid and lipophilic profiles of Chilo partellus (Swinhoe) larvae fluctuate with diapause.

The Chilo partellus (Crambidae: Lepidoptera) larvae undergoes both hibernation and estivation in India. Although, much has been done on reproductive physiological aspects, little is known about biochemical changes happening during hibernation and estivation in C. partellus. Thus, we mapped changes in amino acid and lipophilic profiles of C. partellus larvae while undergoing hibernation and estivation using high-performance liquid chromatography and gas chromatography mass spectroscopy. The studies revealed higher amounts of amino acids namely, serine, glycine, histidine, arginine, proline, tyrosine, and methionine in estivation, while lower in hibernation as compared with nondiapause larvae of C. partellus. Furthermore, the amounts of aspartic acid, glutamic acid, and alanine in hibernation, and threonine, valine, isoleucine, phenylalanine, and leucine in estivation were on par with nondiapause larvae. The lipophilic compounds namely, linoleic acid, stearic acid, eicosanoic acid, and n-pentadecanol were lower in hibernation than estivation and nondiapause larvae of C. partellus. Palmitoleic acid and methyl 3-methoxytetradecanoate contents were higher in hibernation than estivation and nondiapause, while myristic acid and lathosterol contents were higher in estivation than hibernation and nondiapause larvae of C. partellus. Cholesterol content was higher, while squalene and gamma-ergostenol were lower in hibernation and estivation as compared with nondiapause larvae of C. partellus. These findings suggest that certain amino acids may be constituents of heat-shock proteins and help C. partellus during estivation. However, the lipophilic compounds could be helpful in maintaining development during hibernation and estivation in C. partellus.

Biological and biochemical diversity in different biotypes of spotted stem borer, Chilo partellus (Swinhoe) in India.

Because of variation in incidence and severity of damage by Chilo partellus (Swinhoe) in different geographical regions, it is difficult to identify stable sources of resistance against this pest. Therefore, the present studies were undertaken on biological attributes (damage in resistant and susceptible genotypes, survival and development) and biochemical profiles (amino acids and lipophilic compound) of C. partellus populations from eight geographical regions to understand it's population structure in India. There was a significant variation in biological attributes and biochemical profiles of C. partellus populations from different geographical regions. Based on virulence and biological attributes, similarity index placed the C. partellus populations in five groups. Likewise, lipophilic and amino acid profiling also placed the C. partellus populations in five groups. However, the different clusters based on biological and biochemical attributes did not include populations from the same regions. Similarity index based on virulence, biological attributes, and amino acids and lipophilic profiles placed the C. partellus populations in six groups. The C. partellus populations from Hisar, Hyderabad, Parbhani and Coimbatore were distinct from each other, indicating that there are four biotypes of C. partellus in India. The results suggested that sorghum and maize genotypes need to be tested against these four populations to identify stable sources of resistance. However, there is a need for further studies to establish the restriction in gene flow through molecular approaches across geographical regions to establish the distinctiveness of different biotypes of C. partellus in India.

Genetic regulation of diapause and associated traits in Chilo partellus (Swinhoe).

Diapause is an endocrine controlled arrested metabolic state to delay development or reproduction under unfavorable conditions. To gain an understanding of importance of diapause for ecological adaptation, it is important to study regulation of diapause in insects. We examined genetics of diapause in Chilo partellus by crossing the hibernating (HD), aestivating (AD), post-hibernating (PHD), post-aestivating (PAD), and nondiapause (ND) strains. Reciprocal crosses were also made to gain full understanding of diapause regulation and the maternal effects, if any. Data were recorded on fecundity, egg hatching, larval survival, diapause induction and termination, adult emergence, and morphometrics of larvae, pupae and adults in the parents (P1, P2), F1 hybrids, and the reciprocal crosses. Genetic analysis showed that AD strain is general combiner, which also improved egg hatching, larval survival, diapause termination, adult emergence and proportion of females in the progenies. Incidence of diapause was highest in HD × AD, whereas termination was greatest in PHD × AD. However, ND strain and its reciprocal crosses with other strains did not exhibit any noticeable developmental response associated with diapause. Specific combining ability analysis suggested that where PHD and AD strains exist together there will be likely reduction in diapause incidence, increased survival with greater fitness and faster multiplication of their progenies resulting in outbreak of C. partellus. Degree of dominance estimates revealed that diapause, developmental and morphometric traits in C. partellus are governed by over dominance gene effects, and mainly depend on parental diapause history.

Factors responsible for estivation in spotted stem borer, Chilo partellus (Swinhoe).

Diapause is an important adaptive trait in certain groups of insects at a particular stage, which contribute in controlling insect ecology, phenology, and physiology. Considering the importance of various factors in appropriate combinations, we studied effects of different temperature and photoperiod conditions on larval morphometrics, development, induction, and termination of estivation, and overall mortality due to estivation in Chilo partellus (Swinhoe). Weight, length, and head capsule width of C. partellus larvae significantly increased with increase in temperature and photoperiod treatments upto certain age and duration of exposure. According to Dyar's ratio, overall head capsule width progressed geometrically with minimum constant ratio of 1.20, and second-degree polynomial regression equations were best fitted to all the test treatment conditions. The mortality of diapausing C. partellus individuals varied between 7.0% and 37.3% under different constant and varying temperature and photoperiod regimes, which at 38°C + 14L:10D reached upto 100.0%. However, the exposure to 30°C + 12.5L:11.5D and 32°C + 13L:11D along with dry food accelerated the process of estivation, wherein the diapause larval survival was significantly higher and overall mortality of individuals due to diapause was lower suggesting these conditions appropriate for induction of estivation in C. partellus. These studies have implications for better understanding the bioecology and population regulation system under varying climatic conditions and devising novel strategies for management of C. partellus.

Fitness consequences of delayed mating on reproductive performance of Chilo partellus (Swinhoe).

We investigated the effects of mating among different-aged males and females on reproductive physiology, progeny production, and longevity of Chilo partellus (Swinhoe) adults under laboratory conditions. Present studies involved virgin males (M) and females (F) of three different ages namely, on the day of emergence (1D), next day (2D), and further next day (3D), thus comprising of a total of nine mating treatments (1DM × 1DF, 1DM × 2DF, 1DM × 3DF, 2DM × 1DF, 2DM × 2DF, 2DM × 3DF, 3DM × 1DF, 3DM × 2DF, and 3DM × 3DF). The observations were recorded on fecundity, fertility, hatchability, and longevity of male and female adults. We found that mating among old aged males and females significantly reduce fecundity, fertility, and egg hatchability. Further, the fecundity, fertility, and egg hatching were significantly higher when newly emerged females mated with either of the ages of males as compared with other mating treatments. Conversely, the 3-day-old females, when mated with either of the ages of males, laid more numbers of unfertilized eggs, thus reduction in egg hatchability. Mating among newly emerged adults significantly reduced the longevity of male and female adults. Kaplan-Meier estimation showed that none of the adults across mating treatments survived more than 6 days. The recovery of spermatophores from the reproductive tract of females from 3DM × 1DF were significantly higher than other mating treatments. However, no association was found between spermatophore recovery and longevity of male and female adults. Findings of present studies will help to devise appropriate techniques to disrupt mating and suppress C. partellus population under field conditions.