Safe utilization and remediation potential of the mulberry-silkworm system in heavy metal-contaminated lands: A review.

Mulberry cultivation and silkworm rearing hold a prominent position in the agricultural industries of many Asian countries, contributing to economic growth, sustainable development, and cultural heritage preservation. Applying the soil-mulberry-silkworm system (SMSS) to heavy metal (HM)-contaminated areas is significant economically, environmentally, and socially. The ultimate goal of this paper is to review the main research progress of SMSS under HM stress, examining factors affecting its safe utilization and remediation potential for HM-contaminated soils. HM tolerance of mulberry and silkworms relates to their growth stages. Based on the standards for HM contaminants in various mulberry and silkworm products and the bioconcentration factor of HMs at different parts of SMSS, we calculated maximum safe Cd and Pb levels for SMSS application on contaminated lands. Several remediation practices demonstrated mulberry's ability to grow on barren lands, absorb various HMs, while silkworm excreta can adsorb HMs and improve soil fertility. Considering multiple factors influencing HM tolerance and accumulation, we propose a decision model to guide SMSS application in polluted areas. Finally, we discussed the potential of using molecular breeding techniques to screen or develop varieties better suited for HM-contaminated regions. However, actual pollution scenarios are often complex, requiring consideration of multiple factors. More large-scale applications are crucial to enhance the theoretical foundation for applying SMSS in HM pollution risk areas.

Isolation and characterization of bacteria associated with silkworm gut under antibiotic-treated larval feeding / Isolamento e caracterização de bactérias associadas ao trato intestinal do bicho-da-seda sob alimentação larval tratada com antibióticos

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Isolation and characterization of bacteria associated with silkworm gut under antibiotic-treated larval feeding

Abstract The impact of antibiotics on growth, cocoon production was assessed in addition to isolation and characterization of bacteria associated with silkworm gut of infected larvae. Larval rearing was maintained at recommended conditions of temperature and humidity. Silkworm larvae showing abnormal symptoms were collected from the control group and dissected for gut collection. Bacteria were isolated from the gut content by spreading on agar plates and incubated at 37 °C for 48 hrs. Bacterial identification and phylogenetic analysis were carried out by 16S rRNA gene sequencing. The isolated bacteria were subjected to antimicrobial susceptibility test (disc diffusion methods) by using Penicillin (10 µg/mL), Tetracycline (30 µg/mL), Amoxicillin (25 µg/mL), Ampicillin (10 µg/mL), and Erythromycin (15 µg/mL). All isolated strains showed positive results for the catalase test. We isolated and identified bacterial strains (n = 06) from the gut of healthy and diseased silkworm larvae. Based on the 16S rRNA gene sequence, isolated bacteria showed close relation with Serratia, Bacillus, and Pseudomonas spp. Notably, 83.3% of strains were resistant to Penicillin, Tetracycline, Amoxicillin, Ampicillin, and Erythromycin but 16.6% showed antibiotic susceptibility to the above-mentioned commonly used antibiotics. Silkworm larvae fed on penicillin-treated leaves showed significant improvement in larval weight, larval length, and cocoon production. Significantly higher larval weight (6.88g), larval length (5.84cm), and cocoon weight (1.33g) were recorded for larvae fed on leaves treated with penicillin as compared to other antibiotics. Isolated bacterial strains showed close relation with Serratia spp., Bacillus spp. and Pseudomonas spp.

Resumo O impacto dos antibióticos no crescimento e na produção do casulo foi avaliado, além do isolamento e caracterização das bactérias associadas ao intestino de larvas infectadas do bicho-da-seda. A criação das larvas foi mantida nas condições recomendadas de temperatura e umidade. As larvas do bicho-da-seda com sintomas anormais foram coletadas do grupo controle e dissecadas para coleta do intestino. As bactérias foram isoladas do conteúdo intestinal por espalhamento em placas de ágar e incubadas a 37° C durante 48 horas. A identificação bacteriana e a análise filogenética foram realizadas pelo sequenciamento do gene 16S rRNA. As bactérias isoladas foram submetidas a teste de sensibilidade antimicrobiana (métodos de difusão em disco) com penicilina (10 µg / mL), tetraciclina (30 µg / mL), amoxicilina (25 µg / mL), ampicilina (10 µg / mL) e eritromicina (15 µg / mL). Todas as cepas isoladas apresentaram resultados positivos para o teste da catalase. Isolamos e identificamos cepas bacterianas (n = 06) do intestino de larvas de bicho-da-seda saudáveis e doentes. Com base na sequência do gene 16S rRNA, as bactérias isoladas mostraram estreita relação com Serratia, Bacillus e Pseudomonas spp. Notavelmente, 83,3% das cepas eram resistentes a penicilina, tetraciclina, amoxicilina, ampicilina e eritromicina, mas 16,6% mostraram suscetibilidade aos antibióticos comumente usados mencionados acima. As larvas do bicho-da-seda alimentadas com folhas tratadas com penicilina apresentaram melhora significativa no peso larval, comprimento larval e produção de casulo. Peso larval significativamente maior (6,88g), comprimento larval (5,84cm) e peso do casulo (1,33g) foram registrados para larvas alimentadas com folhas tratadas com penicilina, em comparação com outros antibióticos. Cepas bacterianas isoladas mostraram estreita relação com Serratia spp., Bacillus spp. e Pseudomonas spp.

Diversity of fungal pathogens in leaf spot disease of Indian mulberry and its management.

Leaf spot disease in mulberry significantly affects silk production by reducing the nutritive quality of the leaves. This disease caused by various pathogens, regardless of the localities under the same climatic region. In the present investigation, an approximate incidence rate of 84 % was recorded in Karnataka based on surveys conducted in both farmer fields and germplasm locations. The causative agents have shown diversification, including new candidates such as Bipolaris sorokiniana, Curvularia lunata, Cladosporium sphaerospermum, and Epicoccum sorghinum. These findings mark the first report of these pathogens in Indian mulberry production. The investigation involved detailed pathogenicity assessments on the predominant mulberry silk production cultivar under controlled and field environments. Pathogens were identified using morpho-cultural, microscopic and phylogenetic analyses, including the internal transcribed spacer (ITS). Various concentrations of fungicides, both individually and in combinations, were evaluated to identify effective measures for mitigating yield losses. Among the fungicides tested against the new pathogens, Hexaconazole 5 % SC and Hexaconazole 5 % + Captan 70 % WP demonstrated high promise and cost-effectiveness. Consequently, these fungicides could serve as immediate solutions to prevent further yield reduction. However, it is essential to conduct comprehensive field investigations before recommending them as standard practices. Future research endeavors should focus on assessing the extent of crop loss caused by these newly identified pathogens in mulberry cultivation.

Thermal tolerance role of novel polyamine, caldopentamine, identified in fifth instar Bombyx mori.

Silkworms have limited ability to regulate their body temperature; therefore, environmental changes, such as global warming, can adversely affect their viability. Polyamines have shown protection to various organisms against heat stress. This study evaluated the qualitative and quantitative changes in heat-stressed Bombyx mori larvae polyamines. Fifth instar Bombyx mori larvae were divided into two groups; control group, reared at room temperature, i.e., 28 ± 2 °C, and the heat shock group, exposed to 40 °C. Dansylation of the whole worm polyamines and subsequent thin-layer chromatography revealed the presence of components with the same Rf value as dansyl-putrescine, spermidine, and spermine. The dansyl-putrescine, spermidine, and spermine polyamines were identified by mass spectrometric analyses. After heat shock, the thin-layer chromatography of the whole-larvae tissue extracts showed qualitative and quantitative changes in dansylated polyamines. A new polyamine, caldopentamine, was identified, which showed elevated levels in heat-stressed larvae. This polyamine could play a role in helping the larvae tolerate various stress, including thermal stress. No significant changes in silk fiber's economic and mechanical properties were observed in our study. This study indicated that PA, caldopentamine, supplementation could improve heat-stress tolerance in Bombyx mori.

Investigation on Pathological Aspects, Mode of Transmission, and Tissue Tropism of Antheraea proylei Nucleopolyhedrovirus Infecting Oak Tasar Silkworm.

The temperate oak tasar silkworm, Antheraea proylei, is frequently infested with Antheraea proylei nucleopolyhedrovirus (AnprNPV) causing tiger band disease. This disease is one of the key factors that obstructs production and productivity of oak tasar sericulture. The current study aimed to investigate the pathogenicity of AnprNPV, its mode of transmission, and detection of AnprNPV in different tissues. Transmission electron micrographs of AnprNPV showed single rod-shaped bodies and occlusion derived virus (ODV) enclosed within multiple envelopes. The infecting AnprNPV displayed tissue tropism with higher copy numbers detected in the insect fat body and ovary. The virus was observed to multiply in all developmental stages of the silkworm such as egg, larva, pupa, and moth, confirming its ability to spread throughout the silkworm lifecycle. Baculovirus isolated from infected A. proylei showed cross-infectivity in other Saturniidae wild silkworm species such as Antheraea pernyi, A. frithi, and Samia ricini, widening their probable host range for infection. Baculoviruses generally display a horizontal mode of transmission, mainly through ingestion of occlusion bodies (OBs); however, the present study revealed a trans-ovum vertical mode of transmission in addition to a horizontal mode. The observations made in this study aid a detailed understanding of the tiger band disease and its causative pathogen AnprNPV, which will support future studies and disease management in oak tasar sericulture.

Lens on Tropical Sericulture Development in Indonesia: Recent Status and Future Directions for Industry and Social Forestry.

The domestic silkworm or mulberry silk moth, B. mori L., provides more than 99% of the world's silk. Silk, as a sericulture product, was first introduced in Indonesia through a trade mechanism and began to develop in 1953. Several factors (economic, ecological, market, and cultural) support sericulture and make it become one of the non-timber forest product priorities. However, the competitive advantages alone have not encouraged the development of prospective sericulture industry in Indonesia yet. This paper is a review of tropical sericulture development in Indonesia. The literature on the development of sericulture in Indonesia between 1989 and 2022 is used to describe conditions related to mulberry cultivation (moriculture), and silkworm rearing (sericulture), as well as the state of socio-economic development, culture, and institutions. Moriculture and sericulture techniques, socio-economic aspects, institutional arrangements, and community motivations are intertwined, creating a challenging atmosphere for sericulture development. There are potential resources, such as exploring quality mulberry production and quality silkworm production through research and development, valuable cultural aspects, and potential stakeholders to build network engagement. Commitment, cooperation, and action from all stakeholders are needed to enhance the development of sericulture in Indonesia. In this context, the central government can play an important role in facilitating multi-stakeholder partnerships in the development of integrated sericulture in Indonesia.

Pilot study on provenance tracing of cocoons via strontium isotopes.

Isotopic analysis has seen significant use in archaeological sciences to date objects, determine their origin, and depict ancient human dietary habits. However, the potential of this technique for provenance studies of ancient silks remains underdeveloped. In this study, we applied Sr isotopic ratios to the provenance tracing of silkworm cocoons. We investigated the 87Sr/86Sr ratios of cocoons from seven Chinese provinces to determine their regional differences. The 87Sr/86Sr ratios of mulberry leaves and cocoons in Shandong and Sichuan were analyzed and silkworms were cultured at four sampling locations in Hangzhou, Zhejiang, to determine isotopic signature relationships between mulberry leaves and cocoons. Those results showed that the 87Sr/86Sr signature of cocoons not only reflected regional differences, but also was related to the type of soil in each sampling location from which the samples were collected. It is suggested that the Sr isotope ratios was able to be an effective tool for the provenance tracing of cocoons. The Mann-Whitney test result indicated no significant differences in isotopic ratios between mulberry leaves and cocoons. In other words, mulberry leaves may predict mean isotopic values in the cocoons. No clear evidence of Sr isotopic fractionation was found in our control experiments. However, mulberry leaves and cocoons from Sichuan did not show significant correlation between them, overall reducing the predictive power of the 87Sr/86Sr ratios of mulberry leaf for provenance studies of cocoons. Finally, in order to improve the accuracy of Sr isotope ratios for the provenance tracing of cocoons, more 87Sr/86Sr data should be complemented and the relationship needs to be established between Sr isotope information in more kinds of proxies and cocoons.

Bibliometric Analysis of Trends in Mulberry and Silkworm Research on the Production of Silk and Its By-Products.

Traditionally, sericulture is meant as the agricultural activity of silk production, from mulberry (Morus sp.pl.) cultivation to silkworm (Bombyx mori L.) rearing. The aim of the present work is to analyze the trends and outputs of scientific research on sericulture-related topics during the last two decades, from 2000 to 2020. In this work the authors propose a text-mining analysis of the titles, abstracts and keywords of scientific articles focused on sericulture and available in the SCOPUS database considering the above-mentioned period of time; from this article collection, the 100 most recurrent terms were extracted and studied in detail. The number of publications per year in sericulture-related topics increased from 87 in 2000 to 363 in 2020 (+317%). The 100 most recurrent terms were then aggregated in clusters. The analysis shows how in the last period scientific research, besides the traditional themes of sericulture, also focused on alternative products obtainable from the sericultural practice, as fruits of mulberry trees (increment of +134% of the occurrences in the last five years) and chemical compounds as antioxidants (+233% of occurrences), phenolics (+330% of occurrences) and flavonoids (+274% of occurrences). From these considerations, the authors can state how sericulture is an active and multidisciplinary research field.

Transcriptome analysis of resistant and susceptible mulberry responses to Meloidogyne enterolobii infection.

BACKGROUND: Mulberry (Morus alba L.) is an important sericulture crop; however, root-knot nematode infection seriously limits its production. Understanding the mechanism of interaction between mulberry and nematode is important for control of infection. RESULTS: Using sequencing and de novo transcriptome assembly, we identified 55,894 unigenes from root samples of resistant and susceptible mulberry cultivars at different stages after infection with the nematode Meloidogyne enterolobii; 33,987 of these were annotated in the Nr, SWISS-PROT, KEGG, and KOG databases. Gene ontology and pathway enrichment analyses of differentially expressed genes (DEGs) revealed key genes involved in hormone metabolic processes, plant hormone signal transduction, flavonoid biosynthesis, phenylpropanoid biosynthesis, and peroxisomal and photosynthetic pathways. Analysis of key trends in co-expression networks indicated that expression of unigenes 0,015,083, 0,073,272, 0,004,006, and 0,000,628 was positively correlated with resistance to M. enterolobii. Unigene 0015083 encodes tabersonine 16-O-methyltransferase (16OMT), which is involved in alkaloid biosynthesis. Unigene 0073272 encodes a transcription factor contributing to nitric oxide accumulation during plant immune responses. Unigenes 0,004,006 and 0,000,628 encode ERF and MYB transcription factors, respectively, involved in plant hormone signaling. We verified the accuracy of transcriptome sequencing results by RT-qPCR of 21 DEGs. CONCLUSIONS: The results of this study increase our understanding of the resistance mechanisms and candidate genes involved in mulberry-M. enterolobii interaction. Thus, our data will contribute to the development of effective control measures against this pathogen.

Pebrine diagnosis using quantitative phase imaging and machine learning.

Pebrine is the most dreaded infectious disease of the silkworm and has devastated sericulture in Europe during the 18th century. Thereafter, if it is detected, the crop is burned to prevent further dissemination. The conventional microscopic examination of moth's body fluid is erroneous and it exacerbates on Metarhizium anisopliae (MA) contaminated test samples. This is due to the resemblance of pebrine and MA spores in the microscopic examination. Therefore, this study aims to demonstrate an efficient pebrine detection technique. In the proposed method, a motorised brightfield microscope is custom-made to acquire focused and defocused images of test spores. These images are used to produce quantitative phase images of the spores by the transport of intensity equation method. The phase images' histogram of oriented gradients feature is used by a machine learning classifier to categorise the spores. This system classified 92 pebrine and 185 MA spores with an accuracy of 97% at 0.04 seconds/spore. The duration taken for image acquisition is 2.5 minutes per sample (10 fields of view covering an area of 302 × 260 µm2 ). The proposed method shows reliable results in pebrine diagnosis and would be an efficient alternative for current approaches.

Host Response against Virus Infection in an Insect: Bidensovirus Infection Effect on Silkworm (Bombyx mori).

Silk cocoons obtained from silkworms are the primary source of commercial silk, making the silkworm an economically important insect. However, the silk industry suffers significant losses due to various virus infections. Bombyx mori bidensovirus (BmBDV) is one of the pathogens that cause flacherie disease in silkworms. Most silkworm strains die after BmBDV infection. However, certain silkworm strains show resistance to the virus, which is determined by a single recessive gene, nsd-2. The +nsd-2 gene (allele of nsd-2; the susceptibility gene) encodes a putative amino acid transporter expressed only in the insect's midgut, where BmBDV can infect, suggesting that this membrane protein may function as a receptor for BmBDV. Interestingly, the expression analysis revealed no changes in the +nsd-2 gene expression levels in virus-uninfected silkworms, whereas the gene expression drastically decreased in the virus-infected silkworm. This condition indicates that the host factor's expression, the putative virus receptor, is affected by BmBDV infection. It has recently been reported that the expression levels of some host genes encoding cuticle, antioxidant, and immune response-related proteins were significantly regulated by BmBDV infection. In this review, we discuss the host response against virus infection based on our knowledge and long-term research experience in this field.

A review on the biological effects of nanomaterials on silkworm (Bombyx mori).

The production of high-quality silkworm silk is of importance in sericulture in addition to the production of biomass, silk proteins, and animal feed. The distinctive properties of nanomaterials have the potential to improve the development of various sectors including medicine, cosmetics, and agriculture. The application of nanotechnology in sericulture not only improves the survival rate of the silkworm, promotes the growth and development of silkworm, but also improves the quality of silk fiber. Despite the positive contributions of nanomaterials, there are a few concerns regarding the safety of their application to the environment, in humans, and in experimental models. Some studies have shown that some nanomaterials exhibit toxicity to tissues and organs of the silkworm, while other nanomaterials exhibit therapeutic properties. This review summarizes some reports on the biological effects of nanomaterials on silkworm and how the application of nanomaterials improves sericulture.

Black, Caspian Seas and Central Asia Silk Association (BACSA) for the Future of Sericulture in Europe and Central Asia.

The history and recent activities of the Black Caspian Seas and Central Asia Silk Association are presented in this paper: the countries that participated in its foundation, the FAO's action to revitalize sericulture in Eastern Europe and Central Asia, the following widening of the Association geographical limits of to enclose other European countries, which were not well-represented in other similar organizations. Some statistical data are illustrated for a better description of the scenario in which the BACSA executive board acted: the world silk production quantity and the relative production of BACSA countries in respect to the Chinese expansion. The themes treated in the BACSA conferences are reported to explain which matters the Executive Board considered the most relevant for the relaunch of this activity in relationships to the international challenges in the subsequent years; the project proposals that were presented to international donors are summarized. A SWOT (Strengths, Weaknesses, Opportunities, Threats) analysis is shown, where key-factors in determining the strengths and weaknesses of this organization and its member countries for a successful re-establishment of sericulture, are considered. In addition, future trends of sericulture with regard to innovative productions and the Green Deal are examined.

Construction of Baculovirus-Inducible CRISPR/Cas9 Antiviral System Targeting BmNPV in Bombyx mori.

The silkworm Bombyx mori is an economically important insect. The sericulture industry is seriously affected by pathogen infections. Of these pathogens, Bombyx mori nucleopolyhedrovirus (BmNPV) causes approximately 80% of the total economic losses due to pathogen infections. We previously constructed a BmNPV-specific CRISPR/Cas9 silkworm line with significantly enhanced resistance to BmNPV. In order to optimize the resistance properties and minimize its impact on economic traits, we constructed an inducible CRISPR/Cas9 system for use in transgenic silkworms. We used the 39k promoter, which is induced by viral infection, to express Cas9 and the U6 promoter to express four small guide RNA targeting the genes encoding BmNPV late expression factors 1 and 3 (lef-1 and lef-3, respectively), which are essential for viral DNA replication. The system was rapidly activated when the silkworm was infected and showed considerably higher resistance to BmNPV infection than the wild-type silkworm. The inducible system significantly reduced the development effects due to the constitutive expression of Cas9. No obvious differences in developmental processes or economically important characteristics were observed between the resulting transgenic silkworms and wild-type silkworms. Adoption of this accurate and highly efficient inducible CRISPR/Cas9 system targeting BmNPV DNA replication will result in enhanced antivirus measures during sericulture, and our work also provides insights into the broader application of the CRISPR/Cas9 system in the control of infectious diseases and insect pests.

Chromosome-Level Reference Genome and Population Genomic Analysis Provide Insights into the Evolution and Improvement of Domesticated Mulberry (Morus alba).

Mulberry (Morus spp.) is the sole plant consumed by the domesticated silkworm. However, the genome of domesticated mulberry has not yet been sequenced, and the ploidy level of this species remains unclear. Here, we report a high-quality, chromosome-level domesticated mulberry (Morus alba) genome. Analysis of genomic data and karyotype analyses confirmed that M. alba is a diploid with 28 chromosomes (2n = 2x = 28). Population genomic analysis based on resequencing of 134 mulberry accessions classified domesticated mulberry into three geographical groups, namely, Taihu Basin of southeastern China (Hu mulberry), northern and southwestern China, and Japan. Hu mulberry had the lowest nucleotide diversity among these accessions and demonstrated obvious signatures of selection associated with environmental adaptation. Further phylogenetic analysis supports a previous proposal that multiple domesticated mulberry accessions previously classified as different species actually belong to one species. This study expands our understanding of genome evolution of the genus Morus and population structure of domesticated mulberry, which would facilitate mulberry breeding and improvement.

Effect of Honey (Apis dorsata [Hymenoptera: Apidae]) on Larval Growth and Silk Cocoon Yield of Bombyx mori (Lepidoptera: Bombycidae).

The present study was carried out to determine the influence of 2% aqueous honey (Apis dorsata Fabricius, 1793 [Hymenoptera: Apidae]) on larval growth and silk cocoon yield of fifth-instar larvae of the silkworm, Bombyx mori (Linnaeus, 1758) (Lepidoptera: Bombycidae). The larvae of silkworms (Chinese HUAKAND2) were divided into a control and an experimental groups (n = 20 in each group). Control group was fed with plain mulberry leaves throughout the fifth instar, whereas the experimental group was offered mulberry leaves dipped in 2% aqueous solution of honey every other day for 4 d (days 1, 3, 5, and 7). On the other days (days 2, 4, 6, and 8), plain mulberry leaves were offered to larvae. Results showed that the average weight gain in larvae of the experimental group was 348.23 and 204.54% in case of the control group. Uneaten mulberry leaves were weighed; the control group left 34.05% of their leaves and the treated group 28.54%. The cocoon formation in the honey-treated larvae was more uniform in shape than the control group. Furthermore, honey-treated larvae began to form cocoons 7.8 ± 0.23 h earlier than the control group. We also recorded an increase of 15.34% in average weight of cocoons of the experimental group when compared with the control. Average shell percentage of fresh silk cocoons of the control and experimental groups was 20.5 and 23.5%, respectively. It is concluded from the study that 2% aqueous honey has positive impact on the larval growth and cocoon yield of B. mori.

Efecto del compost en la diversidad de bacterias rizosféricas del cultivo de morera (Morus alba) / Effect of compost on the rhizospheric bacteria diversity in mulberry (Morus alba) crops

RESUMEN La producción orgánica de seda incluye la aplicación de compost como practica de cultivo en morera (Morus alba), sin embargo, el efecto de la fertilización orgánica en las poblaciones de bacterias rizosféricas no siempre es positivo. Para evaluar el efecto del compost en la diversidad de bacterias rizosféricas en cultivos de morera (Morus alba), se aplicaron 0, 0.25, 0.5 y 1 kg.m-2 de compost a parcelas con morera dispuestas en un diseño en bloques completos al azar. De cada parcela se extrajo ADN del suelo rizosférico a los 0, 5, 10 y 90 días de aplicado el compost y se amplificó la región V4 del gen ADNr 16S para su secuenciación y asignación taxonómica de los OTUS. Los índices de diversidad alpha mostraron la dominancia de algunos grupos taxonómicos, como los phyla Proteobacteria y Acidobacteria y los géneros Pseudomonas, Opitutus, Luteolibacter y Nitrospir. La diversidad beta indicó similitud entre las muestras influenciadas por la aplicación compost y el incremento de la diversidad en las parcelas muestreadas al final del experimento (90 días). Los grupos taxonómicos dominantes se caracterizan por su función en el ciclo del nitrógeno. Así, se concluyó que la aplicación de 1 kg.m-2 llevó al aumento de la humedad del suelo, el pH y la disponibilidad de nutrientes, lo que incremento la diversidad de bacterias rizosféricas con cambios positivos en composición, riqueza y abundancia en los niveles de orden, familia y género.

ABSTRACT The organic silk production includes the application of compost as cultivation practice to the mulberry crop. However, the effect of the organic fertilization on populations of rhizospheric bacteria is not always positive. To evaluate the effect of compost on the diversity of rhizospheric bacteria in mulberry crops (Morus alba), 0, 0.25, 0.5 and 1 kg.m-2 of compost was applied to mulberry plots arranged in a completely randomized blocks design. From each plot, DNA was extracted from the rhizospheric soil at 0, 5, 10 and 90 days after compost was applied and the V4 region of the 16S rDNA gene was amplified for its sequencing and taxonomic assignment of the OTUS. Alpha diversity indices showed the dominance of some taxonomic groups, such as the phyla Proteobacteria and Acidobacteria, and the genera Pseudomonas, Opitutus, Luteolibacter and Nitrospira. The beta diversity indicated similarity between the samples influenced by the compost application and the increase of the biodiversity in the plots sampled at the end of the experiment (90 days). The taxonomic groups dominated are characterized by their role in the nitrogen cycle. Thus, it was concluded that the application of 1 kg.m-2 of compost led to the increase of ground humidity, soil pH and nutrient availability which increased the rhizospheric bacteria diversity with positive changes in composition, richness and abundance at order, family and genera levels.

Rapid detection of Bombyx mori bidensovirus by loop-mediated isothermal amplification based lateral flow dipstick assay for field applications.

Bombyx mori bidensovirus (BmBDV), is the only bipartite single-stranded DNA (ssDNA) insect virus reported to date. BmBDV causes fatal flacherie disease in silkworm, resulting in large economic losses to sericulture. We developed a loop-mediated isothermal amplification with lateral flow dipstick (LAMP-LFD) method that can successfully and rapidly detect BmBDV DNA with a low limit of detection (5 fg, 400 copies of the BmBDV genome). The method was evaluated and improved for direct field diagnosis using silkworm faeces. In the field, the actual limit of detection was ∼50 fg (4000 copies of the BmBDV genome). The results demonstrated that, compared with traditional methods for BmBDV detection, our new LAMP-LFD method was much more rapid, sensitive and cost-effective, with less dependence on equipment, making it easier to use. The method has potential to be translated into a new diagnostic product for field applications in the sericulture industry.

Role of microsatellites in genetic analysis of Bombyx mori silkworm: a review.

In the genome of Bombyx mori Linnaeus (1758), the microsatellites, or simple sequence repeats (SSR), feature among their particular characteristics a high adenine and thymine (A/T) content, low number of repeats, low frequency, and a grouping in "families" with similar flanking regions. Such characteristics may be the result of a complex interaction between factors that limit the size and dispersion of SSR loci-such as their high association with transposons-and mean that microsatellites within this taxon suitable as molecular markers are relatively rare. The determination of genetic profiles in populations and cell lines has not been affected owing to the high level of polymorphism, nor has the analysis of diversity, structure and genetic relationships. However, the scarcity of suitable microsatellites has restricted their application in genetic mapping, limiting them to preliminary identification of gene location of genes or quantitative trait loci (QTLs) related to thermotolerance, resistance to viruses, pigmentation patterns, body development and the weight of the cocoon, the cortex, the pupa and the filament. The review confirms that, as markers, microsatellites are versatile and perform well. They could thus be useful both to advance research in emerging countries with few resources seeking to promote sericulture in their territories, and to advance in the genetic and molecular knowledge of characteristics of productive and biological interest, given the latest technological developments in terms of the sequencing, identification, isolation and genotyping of SSR loci.