To explore the role of protective enzymes in cassava (Manihot esculenta Crantz) resistance to mites, transgenic cassava lines overproducing copper/zinc superoxide dismutase (MeCu/ZnSOD) and catalase (MeCAT1) were used to evaluate and molecularly confirm cassava resistance to Tetranychus cinnabarinus. Laboratory evaluation demonstrated that, compared with the control cultivar TMS60444 (wild type, WT), the survival, reproduction, development and activities of SOD and CAT in T. cinnabarinus feeding on transgenic cassava lines SC2, SC4, and SC11 significantly inhibited. Furthermore, the activities of SOD and CAT in transgenic cassava lines SC2, SC4, and SC11 damaged by T. cinnabarinus significantly increased. These findings were similar to the results in the mite-resistant cassava cultivars. Besides, field evaluation indicated that the transgenic cassava lines SC2, SC4, and SC11 were slightly damaged as the highly mite-resistant control C1115, while the highly mite-susceptible WT was severely damaged by T. cinnabarinus. Laboratory and field evaluation demonstrated that transgenic cassava lines were resistant to T. cinnabarinus, which directly confirmed that the increase in SOD and CAT activities was positively related to cassava resistance to T. cinnabarinus. These results will help in understanding the antioxidant defense responses in the cassava–mite interaction and molecular breeding of mite-resistant cassava for effective pest control.

To control coconut leaf beetle, Brontispa longissima (Gestro), the pupal parasitoid Tetrastichus brontispae Ferrière was imported from Taiwan and its biology was studied in quarantine in Hainan, China. The parasitoid development includes an egg, three larval instars and three pupal stages. Its developmental time from egg to adult was 19.5±0.5 days under conditions of 24±2°C and 75±5% relative humidity (RH). Temperature had no effect on the sex ratio of offspring, but significantly affected the parasitism rate and reproduction. The parasitism rates were 98.07, 97.97 and 95.03% at 28, 24 and 20°C, respectively, whereas the parasitism rate was 52.18% at 18°C and 69.48% at 30°C, respectively. Furthermore, the parasitoids reared at 18 and 30°C produced fewer offspring than those at 20, 24 and 28°C, respectively. With the increase in temperature, developmental time decreased linearly from 46.19 days at 18°C to 17.10 days at 28°C. RH significantly influenced development, parasitism rate and the reproduction of T. brontispa. With the decrease of RH, developmental time increased from 22.94 days at 20% RH to 18.84 days at 95% RH. In contrast, parasitism rate and the number of offspring per female increased with the increase of RH. Though emergence rates between 50 and 95% RH were much higher than those between 20 and 35% RH, the sex ratios between 20 and 95% RH were not different. Photoperiod had no effect on parasitism, the number of offspring per female, emergence and the sex ratio of T. brontispae, but developmental time was significantly different for different photoperiods. Sucrose, honey and glucose significantly enhanced adult longevity, parasitism and the number of offspring per female of T. brontispae, but had no effect on the sex ratio and survival. Females of T. brontispae only parasitized fourth to fifth larval instars and 1–5-day-old pupae, but there was a significant difference in the number of offspring per female, development time, emergence and the sex ratio of offspring in different instars. These results showed that 1-day-old pupae, a temperature of 24–28°C and 65–95% RH were optimal for T. brontispae. These findings should be helpful in developing a production system to rear and release T. brontispae in large enough quantities to effectively control coconut leaf beetle.

Introduction: The two-spotted spider mite (TSSM) is a devastating pest of cassava production in China. Lignin is considered as an important defensive barrier against pests and diseases, several genes participate in lignin biosynthesis, however, how these genes modulate lignin accumulation in cassava and shape TSSM-resistance is largely unknown.Methods: To fill this knowledge gap, while under TSSM infestation, the cassava lignin biosynthesis related genes were subjected to expression pattern analysis followed by family identification, and genes with significant induction were used for further function exploration.Results: Most genes involved in lignin biosynthesis were up-regulated when the mite-resistant cassava cultivars were infested by TSSM, noticeably, the MePAL gene presented the most vigorous induction among these genes. Therefore, we paid more attention to dissect the function of MePAL gene during cassava-TSSM interaction. Gene family identification showed that there are 6 MePAL members identified in cassava genome, further phylogenetic analysis, gene duplication, cis-elements and conserved motif prediction speculated that these genes may probably contribute to biotic stress responses in cassava. The transcription profile of the 6 MePAL genes in TSSM-resistant cassava cultivar SC9 indicated a universal up-regulation pattern. To further elucidate the potential correlation between MePAL expression and TSSM-resistance, the most strongly induced gene MePAL6 were silenced using virus-induced gene silencing (VIGS) assay, we found that silencing of MePAL6 in SC9 not only simultaneously suppressed the expression of other lignin biosynthesis genes such as 4-coumarate--CoA ligase (4CL), hydroxycinnamoyltransferase (HCT) and cinnamoyl-CoA reductase (CCR), but also resulted in decrease of lignin content. Ultimately, the suppression of MePAL6 in SC9 can lead to significant deterioration of TSSM-resistance.Discussion: This study accurately identified MePAL6 as critical genes in conferring cassava resistance to TSSM, which could be considered as promising marker gene for evaluating cassava resistance to insect pest.

The two-spotted spider mite (TSSM) is a destructive cassava pest. Intensive demonstration of resistance mechanism greatly facilitates the creation of TSSM-resistant cassava germplasm. Gene to metabolite network plays a crucial role in modulating plant resistance, but little is known about the genes and related metabolites which are responsible for cassava resistance to TSSM. Here, a highly resistant (HR) and a highly susceptible (HS) cassava cultivar were used, integrative and comparative transcriptomic and metabolomic analyses between these two cultivars after TSSM infestation revealed that several genes and metabolites were closely related and significantly different in abundance. In particular, the expression of leucoanthocyanidin reductase (LAR) and anthocyanidin reductase (ANR) genes showed a high positive correlation with most of the metabolites in the tannin biosynthesis pathway. Furthermore, transgenic cassava lines overexpressing either of the genes elevated tannin concentrations and conferred cassava resistance to TSSM. Additionally, different forms of tannins possessed distinct bioactivity on TSSM, of which total condensed tannins (LC₅₀ = 375.68 mg/l) showed maximum lethal effects followed by procyanidin B1 (LC₅₀ = 3537.10 mg/l). This study accurately targets LAR, ANR and specific tannin compounds as critical genes and metabolites in shaping cassava resistance to TSSM, which could be considered as biomarkers for evaluation and creation of pest-resistant cassava germplasm.