Nutritional and Feeding Adaptability of Clanis bilineata tsingtauica Larvae to Different Cultivars of Soybean, (Glycine max).

The larvae of Clanis bilineata tsingtauica, a special species of Chinese edible insect, are of great nutritional, medicinal and economic value to humans. This study aimed to clarify the effect of different soybean varieties (Guandou-3 (G3), Ruidou-1 (R1), September cold (SC)) on the nutritional quality and feeding selection behavior of C. bilineata tsingtauica larvae. The results showed that soybean isoleucine (Ile) and phenylalanine (Phe) were positively correlated with larval host selection (HS) and protein content. The order of soybean plants selected by C. bilineata tsingtauica larvae was R1 > SC > G3, and they selected R1 significantly higher than SC and G3 by 50.55% and 109.01%, respectively. The protein content of the larvae fed on R1 was also the highest among the three cultivars. In addition, a total of 17 volatiles belonging to 5 classes were detected from soybeans: aldehydes, esters, alcohols, ketones, and heterocyclic compounds. Pearson's analysis showed that soybean methyl salicylate was positively correlated with larval HS and their protein content, and soybean 3-octenol was negatively correlated with larval HS and their palmitic acid content. In conclusion, C. bilineata tsingtauica larvae are more adapted to R1 than to the other two soybean species. This study provides a theoretical basis for the production of more protein-rich C. bilineata tsingtauica in the food industry.

TM7 (Saccharibacteria) regulates the synthesis of linolelaidic acid and tricosanoic acid, and alters the key metabolites in diapause Clanis bilineata tsingtauica.

Good exploitation and utilization of edible insects can effectively alleviate global food security crisis in years. The study on diapause larvae of Clanis bilineata tsingtauica (DLC) was conducted to explore how gut microbiota regulate the nutrients synthesis and metabolism of edible insects. The results showed that C. bilineata tsingtauica maintained a total and stable nutrition levels at early phase of diapause. The activity of instetinal enzymes in DLC fluctuated markedly with diapause time. Additionally, Proteobacteria and Firmicutes were the predominant taxa, and TM7 (Saccharibacteria) was the marker species of gut microbiota in DLC. Combined the gene function prediction analysis with Pearson correlation analysis, TM7 in DLC was mainly involved in the biosynthesis of diapause-induced differential fatty acids, i.e., linolelaidic acid (LA) and tricosanoic acid (TA), which was probably regulated by changing the activity of protease and trehalase, respectively. Moreover, according to the non-target metabolomics, TM7 might regulate the significant differential metabolites, i.e., D-glutamine, N-acetyl-d-glucosamine and trehalose, via the metabolism of amino acid and carbohydrate pathways. These results suggest that TM7 increased LA and decreased TA via the intestinal enzymes, and altered intestinal metabolites via the metabolism pathways, maybe a key mechanism for regulating the nutrients synthesis and metabolisms in DLC.

Effects of Host Plants on Bacterial Community Structure in Larvae Midgut of Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda, is one of the most important invasive species and causes great damage to various host crops in China. In this study, the diversity and function of gut bacteria in the 5th instar larvae of FAW fed on maize, wheat, potato and tobacco leaves were analyzed through 16S rRNA sequencing. A total of 1324.25 ± 199.73, 1313.5 ± 74.87, 1873.00 ± 190.66 and 1435.25 ± 139.87 operational taxonomic units (OTUs) from the gut of FAW fed on these four different host plants were detected, respectively. Firmicutes, Proteobacteria and Bacteroidetes were the most abundant bacterial phyla. Beta diversity analysis showed that the gut bacterial community structure of larvae fed on different host plants was significantly differentiated. At the genus level, the abundance of Enterococcus in larvae fed on wheat was significantly lower than those fed on the other three host plants. Enterobacter and ZOR0006 were dominant in FAW fed on tobacco leaves, and in low abundance in larvae fed on wheat. Interestingly, when fed on Solanaceae (tobacco and potato) leaves which contained relative higher levels of toxic secondary metabolites than Gramineae (wheat and maize), the genera Enterococcus, Enterobacter and Acinetobacter were significantly enriched. The results indicated that gut bacteria were related to the detoxification and adaptation of toxic secondary metabolites of host plants in FAW. Further analysis showed that replication, repair and nucleotide metabolism functions were enriched in the gut bacteria of larvae fed on tobacco and potato. In conclusion, the gut bacterial diversity and community composition in FAW larvae fed on different host plants showed significant differences, and the insect is likely to regulate their gut bacteria for adaptation to different host plants.

Genomic and Transcriptomic Analysis Reveals Cuticular Protein Genes Responding to Different Insecticides in Fall Armyworm Spodoptera frugiperda.

The fall armyworm (FAW), Spodoptera frugiperda, is a serious pest of crucial crops causing great threats to the food security of the world. It has evolved resistance to various insecticides, while the underlying molecular mechanisms remain largely unknown. Cuticular proteins (CPs), as primary components in cuticle, play an important role in insects' protection against environmental stresses. Few of them have been documented as participating in insecticide resistance in several insect species. In order to explore whether CP genes of the FAW exhibit a functional role in responding to insecticides stress, a total of 206 CPs, classified into eight families, were identified from the genome of the FAW through a homology-based approach coupled with manual efforts. The temporal expression profiles of all identified CP genes across developmental stages and their responses to 23 different insecticides were analyzed using the RNA-seq data. Expression profiling indicated that most of the CP genes displayed stage-specific expression patterns. It was found that the expression of 51 CP genes significantly changed after 48 h exposure to 17 different insecticides. The expression of eight CP genes responding to four insecticides were confirmed by RT-PCR analysis. The results showed that their overall expression profiles were consistent with RNA-seq analysis. The findings provide a basis for further functional investigation of CPs implied in insecticide stress in FAW.

[Root growth characteristics and competitive effects of Ageratina adenophora and four functional type herbaceous plants].

A field experiment was conducted to explore the root competitive effects of Ageratina adenophora and Setaria sphacelata, S. yunnanensis, Eupatorium fortunei, Chenopodium serotinum in monoculture and mixture, and the relative competitive abilities were evaluated. The results showed that the root length, superficial area and volume of A. adenophora in mixture were lower than in monoculture, but those of S. sphacelata were higher in mixture than in monoculture. The biomass of A. adenophora in mixture decreased by 77.1% and that of S. sphacelata increased by 80.4% compared with those in monoculture. The relative yield and competitive balance index of S. sphacelata were significantly higher than those of A. adenophora, and the relative yield was about 1.0, suggesting that the underground competitive ability of S. sphacelata was higher than A. adenophora. The root morphology of S. yunnanensis in monoculture and mixture was higher than those of A. adenophora, but the root morphology of two species in mixture was lower than in monoculture. The biomass of A. adenophora and S. yunnanensis in mixture decreased by 45.3% and 22.8% compared with those in monoculture, respectively. Competition effect parameters showed that A. adenophora was a mutual antagonism with S. yunnanensis. The root morphology of E. fortunei and A. adenophora in mixture showed no significant difference compared with that in monoculture. The biomass of A. adenophora and E. fortunei was lower than that in monoculture, respectively. Competition effect parameters showed that A. adenophora was a superior competitor. In the mixture of A. adenophora and C. serotinum, the root morphology parameters and competitive ability of A. adenophora were superior to those of C. serotinum. Above all, S. sphacelata is a preference plant material to control the A. adenophorum invasion and recover biodiversity in A. adenophorum invasion fields.

Genetic diversity of the planthopper, Sogatella furcifera in the Greater Mekong Subregion detected by inter-simple sequence repeats (ISSR) markers.

The white backed planthopper, Sogatella furcifera (Hemiptera: Delphacidae), is a serious pest of rice in Asia. In the present study, inter-simple sequence repeat (ISSR) markers were employed to investigate the genetic diversity and differentiation of 47 populations sampled from 14 prefectures of the Greater Mekong Subregion. A total of 14 selected primers yielded 121 bright and discernible bands, with an average of 8.6 bands per primer. According to the hierarchical analysis of molecular variance (AMOVA), the genetic variation among geographic regions (79.84%) was higher than that of among populations within region (20.16%), and the FST value was 0.72, indicating a high level of genetic differentiation. Neighbor-Joining cluster analysis of the 47 populations showed two major clusters, one consisting of mostly southwestern Yunnan Province and Myanmar populations; and the other one consisting of southeastern and central of Yunnan Province plus Vietnam and Laos populations. No significant positive correlation was observed between genetic and geographic distances by Mantel test (r = 0.2230, p = 0.8448), indicating the role of geographic isolation did not shape the genetic structure of the sampled S. furcifera populations. This paper provides useful data for understanding and speculating the migration of S. furcifera and reveals available information to develop sustainable strategies for manage this long-range migratory pest.

[Application of ISSR molecular marker in invasive plant species study].

Alien species invasion is one of the most important drivers of worldwide environmental change, which may result in environmental degradation, biodiversity loss, and food and water shortage. It may also increase the possibility and severity of natural disasters, and damage international trade and benefits. In last two decades, DNA-based molecular markers were widely used to detect the genetic diversity of invaded alien species. Inter-simple sequence repeat (ISSR) is a microsatellite-based technique, with the superiorities of simple, quick, reliable, and generating higher levels of DNA polymorphism, and being used as a new molecular marker for genetic study. This paper introduced the principles, characteristics and procedures of ISSR, and summarized its applications in studying the genetic structure, genetic diversity, origin, distribution mode, phylogenesis, and breeding features of invasive plants.

[Genetic variations among populations of Ageratina adenophora from different areas, investigated by inter-simple sequence repeat markers].

Crofton weed, Ageratina adenophora,a troublesome weed in the world,is an invasive organism in China. Genetic diversity of thirty-two Chinese populations of A. adenophora from different areas was analyzed with the inter-simple sequence repeat (ISSR) technique. It indicated that its genetic diversity was rich. Nei's gene diversity and Shannon's information index were 0.235 and 0.372 at species level respectively. Genetic variation of Crofton weed mainly existed in the population, and about 34.5% of the total variance was attributable to population divergence and 65.5% to individual differences within populations. The Mantel Z-statistic test showed that the genetic distance between populations generally increased with geographic distance (r=0.542,p< 0.001),which indicated that isolation by distance is one of the blocks of gene flow of A. adenophora. Genetic diversity level of Crofton weed had the trend to decrease along with the increased altitude (r=0.368,P<0.001), and the mean of Nei's gene diversity and Shannon's diversity indices both decreased with the increased altitude.