Due to increased use of agrochemicals and growing concerns about ecotoxicology, the development of new insecticides, moving away from those with neurotoxic and broad spectrum effects towards insecticides that are safer for the environment and nontarget beneficial species, has been a research priority. Novaluron stands out among these newer insecticides, is an insect growth regulator that is used for the control of insect pests in crops grown close to mulberry plantations. Mulberry serves as food for the silkworm Bombyx mori, which is a nontarget insect of great economic importance to silk production. We investigated the lethal and sublethal effects of Novaluron on the development of B. mori. Larvae were segregated into experimental groups: the control groups (CGs) and the treatment groups (TGs), which were treated with the Novaluron concentration of 0.15 mL/L. Following exposure, we analyzed: larval mortality, changes in the insect life cicle and cytotoxic effects on the midgut cells. This is the first report about the Novaluron’s effects on B.mori. We detected rupture in the integument, complete cessation of feeding, late development, incomplete ecdysis and production of defective cocoons. After 240 h of exposure, there was 100% mortality in TG larvae exposed in the 3rd instar and 20% mortality from larvae exposed in the 5th instar. Cytotoxic effects was observed, such as dilation of cells, emission of cytoplasmic protrusions, extreme rarefaction of the cytoplasm and nuclei, dilation of the endoplasmic reticulum in addition to changes in mitochondria, the presence of large digestive vacuoles and intercellular spaces and the presence of active caspase. Novaluron exposure impairs the midgut and may affect the physiological functions of this organ. Novaluron additionally compromises several phases of insect development, indicating the importance of toxicology studies that utilize different life stages of nontarget species to evaluate the safe use of insecticides.

Guadipyr, which combines neonicotinoid and semicarbazone functional groups in one molecule, exhibits good activity on several pests and high acute and chronic toxicity to silkworms (Bombyx mori). In this report, the effects of low-dose guadipyr on the midgut microbiota and immune system of silkworms were studied. Results showed that the structure and richness of the midgut microbiota of silkworms were altered after being treated with 5.25 mg/L (1/10 of LC₅₀) of guadipyr. The abundance of Pseudomonas was evidently increased, whereas Curvibacter was substantially reduced, which might be related to the growth and immunity of silkworms. The expression of key genes in the Toll, IMD, and JAK/STAT pathways, which ultimately led to the downregulation of antimicrobial peptide genes (AMPs), such as CecA, Defensin1, Leb, and glv2, was reduced upon guadipyr exposure. Simultaneously, the suppression of steroid hormone 20-hydroxyecdysone receptor and response genes, such as BR-C Z4, was detected in the exposed groups. The decreased expression of these immune regulatory pathway-related and 20-hydroxyecdysone signal pathway-related genes indicated that the immune system of silkworms was affected by low-dose guadipyr. Our results revealed the negative effects of guadipyr on silkworms and highlighted the unneglectable toxicity of low-dose guadipyr to this economic insect. Given the risk, it is necessary to control the application of guadipyr in or around the mulberry fields.

The purpose of the current study was to determine the appropriate genotype and concentration of biosynthesized silver nanoparticles effectual in preserving mulberry leaves at the postharvest stage. The preservative effect of silver nanoparticles was determined by their potentiality to prevent xylem blockage, chlorophyll content retention and inhibition of microbial proliferation within a preservative solution. For synthesizing silver nanoparticles, a blend of 10⁻³ M silver nitrate and S1 genotype of the mulberry leaf was found to be most effective. Silver nanoparticles at 6 ppm were observed to be the least effective concentration for preserving mulberry leaves for at least 7 days at the postharvest stage, as evident from physical texture and retention of chlorophyll content. Biosynthesized silver nanoparticles showed negative microbial count during the course of preservation as evident from no colony-forming unit (CFU) until the last day of preservation, while conventional preservative silver nitrate showed traces of CFU on a nutrient agar plate. Besides, these leaves preserved in nanosilver solution showed an almost negligible number of xylem blockage in the petiole, almost equivalent to the blockage nature of fresh leaves caused by the deposition of macromolecules like protein, lignin and suberin. Nanosilver- and silver nitrate–preserved leaves also displayed insignificant accumulation of reactive oxygen species (ROS) and greater retention of membrane integrity than leaves preserved in normal distilled water. Nanosilver solution showed greater durability of preserving mulberry leaves than conventional floral preservative silver nitrate, useful for feeding silkworm larvae during the rainy season.

Phytoremediation has been proven to be an environmentally sound alternative for the recovery of contaminated soils, and the economic profit that comes along with the process might stimulate its field use. This study investigated cadmium (Cd) transfer and detoxification mechanisms in a soil–mulberry–silkworm system to estimate the suitability of the mulberry and silkworm as an alternative method for the remediation of Cd-polluted soil; it also explored the underlying mechanisms regulating the trophic transfer of Cd. The results show that both the mulberry and silkworm have high Cd tolerance. The transfer factor suggests that the mulberry has high potential for Cd extraction from polluted soil. The subcellular distribution and chemical forms of Cd in mulberry leaves show that cell wall deposition and vacuolar compartmentalization play important role in Cd tolerance. In the presence of increasing Cd concentrations in silkworm food, detoxification mechanisms (excretion and homeostasis) were activated so that excess Cd was excreted in fecal balls, and metallothionein levels in the mid-gut, the posterior of the silk gland, and the fat body of silkworms were enhanced. And, the Cd concentrations in silk are at a low level, ranging from 0.02 to 0.21 mg kg⁻¹. Therefore, these mechanisms of detoxification can regulate Cd trophic transfer, and mulberry planting and silkworm breeding has high phytoremediation potential for Cd-contaminated soil.

A large number of defective mulberries are discarded each year because mulberries are easy to break. The red pigments from defective mulberries are recognized as the sustainable sources of anthocyanins extracted from nature. Cyanidin-3-O-rutinoside and cyanidin-3-O-glucoside are the main components of mulberry red pigments, accounting for 50% and 40% of the total, respectively. Cyanidin-3-O-glucoside exhibits anticancer, hypoglycemic, and liver and visceral protection properties. Cyanidin-3-O-glucoside can be prepared by enzymatically hydrolyzing the rhamnosidase bond of cyanidin-3-O-rutinoside. To obtain mulberry red pigment with a high purity of cyanidin-3-O-glucoside, immobilized α-L-rhamnosidase was added to the aqueous two-phase system to construct a liquid-liquid-solid three-phase enzyme catalytic system. After optimization, the three-phase system was composed of 27.12% (w/w) ethanol, 18.10% (w/w) ammonium sulfate, 15% (w/w) mulberry juice, 4.24% (w/w) immobilized α-L-rhamnosidase, and 35.54% (w/w) pure water. The three-phase system was employed to enrich and purify cyanidin-3-O-glucoside at pH 5 and 45 °C for 1 h. The purity of cyanidin-3-O-glucoside was increased from 40 to 82.42% with cyanidin-3-O-rutinoside conversion of 60.68%. The immobilized α-L-rhamnosidase could be reused seven times, maintaining a relative activity of over 50%. Overall, the developed system provided an efficient and simple approach for high purity mulberry red pigment production and recycling in the field of sustainable agriculture. Graphical abstract

Intercropping of arsenic (As) hyperaccumulator Pteris vittata and cash crop Morus alba could improve the As concentration in the hyperaccumulator but decrease As concentration in the intercropped crop. The effects of several amendments on the transfer of As were investigated to determine an enhancement strategy for the intercropping system of P. vittata and M. alba. Phosphorus, in the form of Ca(H₂PO₄)₂, promoted the release of As to the soil solution and apparently increased the As removal from the soil by 42% compared with the untreated variant. The addition of FeSO₄ and CaCO₃ decreased As concentration in the soil solution and the uptake of As by both plant species. The As levels in the mulberry leaves remained under the threshold limits of feedstuffs in China. Intercropping was confirmed as an applicable strategy to manage contaminated soil. Hence, under the condition that all treatments produced safe mulberry leaves, Ca(H₂PO₄)₂ was the appropriate amendment to achieve the highest As removal rate, whereas FeSO₄ could lower the risk of As to further migrate to another medium.

China's Sloping Land Conversion Program (SLCP) was designed to restore perennial plant cover on sloping land in western China, in part to protect the Three Gorges Reservoir (TGR). In this study, we examined use of white mulberry (Morus alba L.) in the SLCP to protect water quality and conserve soil. We established nine runoff monitoring plots divided among three categories (vegetable farming, fallow control, and mulberry plantation) on a bank of the Liangtan River situated at the western margin of the TGR. The land had been used previously by farmers for growing vegetables. We found that soil loss and surface water runoff were lowest in the mulberry plots and highest in the vegetable plots. We used inductively coupled plasma atomic emission spectroscopy (ICP-AES) to assess the concentration of selected heavy metal pollution indicators (Zn, Hg, As, Ni, Pb, Cr, Cd, and Cu) in the monitoring plot soils at the beginning of the experiment in May 2009. The heavy metals were assessed again at the end of the experiment in October 2012, and we found that the concentrations of these pollutants had been reduced in all fallow and mulberry plots, and to the greatest extent in the mulberry plots. We found that levels of Hg, Pb, and Cu increased in the vegetable plots. For these reasons, we conclude that riparian mulberry plantations are useful for reducing rapid runoff of storm water, conserving soil, and sequestering heavy metal pollutants in the TGR region.

A 3-year field experiment was conducted to evaluate the accumulation ability of 3 forage mulberry (Morus atropurpurea Roxb.) varieties (Guisangyou 62, Guisangyou 12, Yuesang 11) for Cadmium (Cd). The results showed that the shoot biomass of forage mulberry could reach to 64.51~69.58 t/ha in 2018. The total production of Guisangyou 12 was the highest among the tested 3 cultivars in each year. However, the biomass of the 3 varieties displayed without significant differences at every sampling time. In addition, the concentrations of Cd in the roots of the 3 mulberry cultivars were higher than those in the corresponding shoots in 2018. But in 2019, the Cd content in roots was near to that of shoots. The Cd concentrations in the roots of the forage mulberry showed Guisangyou 62 > Yuesang 11 > Guisangyou 12. And the Cd concentrations in shoots of all the 3 tested forage mulberry varieties were less than 1 mg/kg, meeting the hygienical standard for feeds (China). Though the 3 forage mulberry varieties were not hyperaccumulators for Cd, they could be used as alternative plants to make use of the Cd-polluted (< 1.0 mg/kg) paddy soils.

Our study aimed to uncover the functions of two species of arbuscular mycorrhizal fungi (AMF) in soil aggregation and plant water content regulation under wetting–drying climate conditions. The climatic characteristics of seasonal drought in karst areas were simulated. Two watering periods were established in a controlled greenhouse to compare the different effects of two genetically different AMF species (Funneliformis mosseae and Rhizophagus intraradices) on the water content of 90-day-old mulberry seedlings and on soil aggregates. Our results showed that inoculation with the Rhizophagus intraradices (R.i) strain was more effective at improving mulberry growth performance than Funneliformis mosseae (F.m) inoculation under semiarid conditions. The AMF remained highly infective and continuously increased the proportion of soil macroaggregates under soil drought stress. As a result, our study showed the potential of AMF to promote sustainable mulberry plantations and the rehabilitation of degraded soil in karst areas of southwestern China.

Insecticides with different modes of action may act in combination, in ways such as drifting, spray equipment residual, or utilizing concurrently in mulberry orchards or nearby agricultural fields. Silkworms may suffer from a diverse impact on the survival. In this study, the toxicity of chlorantraniliprole, lambda-cyhalothrin, and imidacloprid and their combinations to the second instar of silkworms (Bombyx mori (L.)(Lepidoptera: Bombycidae)) were evaluated after 48 and 72 h treatment by the leaf-dipping method and the combination index (CI)-isobologram equation. After 48 h treatment, results indicated that (1) the increasing order of toxicity was imidacloprid < chlorantraniliprole < lambda-cyhalothrin, and that (2) synergism was predominated in most combinations excepted for the lambda-cyhalothrin + imidacloprid combination which displayed an additive effect at fₐ value 0.5. Then, after 72 h treatment, results exhibited that (1) the increasing order of toxicity was imidacloprid < lambda-cyhalothrin < chlorantraniliprole, and that (2) only the chlorantraniliprole + imidacloprid mixture yielded antagonism at fₐ value 0.5; the other combinations performed an additive effect at least. Consequently, combined toxicity of mixtures may pose a worse effect on silkworm than single toxicity of insecticides. Therefore, we suggest that insecticide mixtures should be added into ecotoxicological risk assessment.