Significant progress has been achieved in silkworm (Bombyx mori) research since the last review on this insect was published in this journal in 2005. In this article, we review the new and exciting progress and discoveries that have been made in B. mori during the past 10 years, which include the construction of a fine genome sequence and a genetic variation map, the evolution of genomes, the advent of functional genomics, the genetic basis of silk production, metamorphic development, immune response, and the advances in genetic manipulation. These advances, which were accelerated by the genome sequencing project, have promoted B. mori as a model organism not only for lepidopterans but also for general biology.

Advances in Silkworm Studies Accelerated by the Genome Sequencing of Bombyx mori

Animals synthesize melanin pigments for the coloration of their skin and use it for their protection from harmful solar radiation. Insects use melanins even more ingeniously than mammals and employ them for exoskeletal pigmentation, cuticular hardening, wound healing and innate immune responses. In this review, we discuss the biochemistry of melanogenesis process occurring in higher animals and insects. A special attention is given to number of aspects that are not previously brought to light: (1) the molecular mechanism of dopachrome conversion that leads to the production of two different dihydroxyindoles; (2) the role of catecholamine derivatives other than dopa in melanin production in animals; (3) the critical parts played by various biosynthetic enzymes associated with insect melanogenesis; and (4) the presence of a number of important gaps in both melanogenic and sclerotinogenic pathways. Additionally, importance of the melanogenic process in insect physiology especially in the sclerotization of their exoskeleton, wound healing reactions and innate immune responses is highlighted. The comparative biochemistry of melanization with sclerotization is also discussed.

https://www.mdpi.com/1422-0067/17/10/1753/pdf

Critical Analysis of the Melanogenic Pathway in Insects and Higher Animals

Adult beetles (Coleoptera) are covered primarily by a hard exoskeleton or cuticle. For example, the beetle elytron is a cuticle-rich highly modified forewing structure that shields the underlying hindwing and dorsal body surface from a variety of harmful environmental factors by acting as an armor plate. The elytron comes in a variety of colors and shapes depending on the coleopteran species. As in many other insect species, the cuticular tanning pathway begins with tyrosine and is responsible for production of a variety of melanin-like and other types of pigments. Tanning metabolism involves quinones and quinone methides, which also act as protein cross-linking agents for cuticle sclerotization. Electron microscopic analyses of rigid cuticles of the red flour beetle, Tribolium castaneum, have revealed not only numerous horizontal chitin-protein laminae but also vertically oriented columnar structures called pore canal fibers. This structural architecture together with tyrosine metabolism for cuticle tanning is likely to contribute to the rigidity and coloration of the beetle exoskeleton.

Cuticle formation and pigmentation in beetles

Despite the variety, prominence, and adaptive significance of butterfly wing patterns, surprisingly little is known about the genetic basis of wing color diversity. Even though there is intense interest in wing pattern evolution and development, the technical challenge of genetically manipulating butterflies has slowed efforts to functionally characterize color pattern development genes. To identify candidate wing pigmentation genes, we used RNA sequencing to characterize transcription across multiple stages of butterfly wing development, and between different color pattern elements, in the painted lady butterfly Vanessa cardui. This allowed us to pinpoint genes specifically associated with red and black pigment patterns. To test the functions of a subset of genes associated with presumptive melanin pigmentation, we used clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 genome editing in four different butterfly genera. pale, Ddc, and yellow knockouts displayed reduction of melanin pigmentation, consistent with previous findings in other insects. Interestingly, however, yellow-d, ebony, and black knockouts revealed that these genes have localized effects on tuning the color of red, brown, and ochre pattern elements. These results point to previously undescribed mechanisms for modulating the color of specific wing pattern elements in butterflies, and provide an expanded portrait of the insect melanin pathway.

https://www.genetics.org/content/genetics/205/4/1537.full.pdf

Genetic Basis of Melanin Pigmentation in Butterfly Wings.

Melanin Pathway Genes Regulate Color and Morphology of Butterfly Wing Scales

Pigmentation patterning has long interested biologists, integrating topics in ecology, development, genetics, and physiology. Wildtype neonatal larvae of the silkworm,Bombyx mori, are completely black. By contrast, the epidermis and head of larvae of the homozygous recessive sex-linked chocolate (sch) mutant are reddish brown. When incubated at 30 °C, mutants with the sch allele fail to hatch; moreover, homozygous mutants carrying the allele sch lethal (sch l ) do not hatch even at room temperature (25 °C). By positional cloning, we narrowed a region containing sch to 239,622 bp on chromosome 1 using 4,501 backcross (BC1) individuals. Based on expression analyses, the best sch candidate gene was shown to be tyrosine hydroxylase (BmTh). BmTh coding sequences were identical among sch, sch l , and wild-type. However, in sch the ∼70-kb sequence was replaced with ∼4.6 kb of a Tc1mariner type transposon located ∼6 kb upstream of BmTh, and in sch l , a large fragment of an L1Bm retrotransposon was inserted just in front of the transcription start site of BmTh. In both cases, we observed a drastic reduction of BmTh expression. Use of RNAi with BmTh prevented pigmentation and hatching, and feeding of a tyrosine hydroxylase inhibitor also suppressed larval pigmentation in the wild-type strain, pnd + and in a pS (black-striped) heterozygote. Feeding L-dopa to sch neonate larvae rescued the mutant phenotype from chocolate to black. Our results indicate the BmTh gene is responsible for the sch mutation, which plays an important role in melanin synthesis producing neonatal larval color.

/pdf/repression-of-tyrosine-hydroxylase-is-responsible-for-the-z5pm2z32h3.pdf

Yu Han

Papers

Repression of tyrosine hydroxylase is responsible for the sex-linked chocolate mutation of the silkworm, Bombyx mori