Insect cuticle tanning (pigmentation and sclerotization) is a complex and vital physiological process that begins with tyrosine and is responsible for production of both melanin- and quinoid-type pigments. In addition, these quinones undergo isomerization to quinone methides and cross-linking reactions with cuticular proteins for cuticle sclerotization. In this study, we studied the functions of TmDDC and TmY-y as well as TmNAT1, TmADC and Tmebony from Tenebrio molitor, which are involved in the tyrosine-derived melanin- and quinoid-type pigment productions, respectively. The temporal and spatial expression patterns of the genes were analyzed by real-time PCR. RNA interference was performed to understand the genetic regulation and molecular mechanism underlying the darkening and hardening of beetle cuticle.

Chitin deacetylases (CDAs) are extracellular-modifying enzymes that deacetylate chitin to produce chitosan. In insects, this modification may contribute to the affinity and/or cross-linking of chitin/chitosan-like polysaccharides for a variety of structural proteins, which may lead to diverse mechanical properties of the cuticle. We previously reported the functional importance of Group I CDAs, TcCDA1 and TcCDA2, as well as the two alternative spliced isoforms of the latter, TcCDA2a and TcCDA2b from the red flour beetle, Tribolium castaneum in molting, morphology of cuticle and movement of legs. In this study, we further analyzed protein localization, ultrastructural defects of the cuticles and leg joints after RNAi of those genes. Both proteins are mainly present in the innermost procuticle region called the “assembly zone”. Loss of function of either TcCDA1 or TcCDA2 caused disorganized chitinous horizontal laminae and vertical pore canals in both the rigid and soft cuticles. RNAi of TcCDA2b affects cuticle integrity similar to that seen in RNAi of the two alternatively spliced forms of TcCDA2. In contrast, TcCDA2a-deficient adult, like that seen in the hypomorphic phenotype produced by RNAi of TcCDA1, exhibited ruptured tendons between femur and tibia, resulting in loss of locomotion ability. These results suggest that Group I CDAs play critical roles in molting, morphology, ultrastructure and mobility in T. castaneum. This work was supported by NRFs (NRF-2015R1A6A3A04060323 and NRF-2018R1A2B6005106).

Insect cuticle tanning (pigmentation and sclerotization) is a complex and vital process, which includes hydroxylation of initial amino acid, tyrosine, to DOPA and decarboxylation of DOPA to dopamine. In the pigmentation process, dopamine further undergoes two N-acylation reactions to yield N-acetyldopamine (NADA) and N-β-alanyldopamine (NBAD). In the former reaction, arylalkylamine N-acetyltransferase (AANAT1) converts dopamine to NADA, and in the later reaction, aspartate 1-decarboxylase (ADC) provides β-alanine, which is conjugated with dopamine catalyzed by NBAD synthase (Ebony) for production of NBAD. In this study, we performed functional genomics of TmAANAT1, TmADC and Tmebony to determine whether they are required for cuticle pigmentation in Tenebrio molitor adults. Loss of function of these genes by RNAi caused the significantly darker body color than that of control animals. Note that, although all phenotypes exhibited dark cuticle pigmentation, RNAi of either TmADC or Tmebony only altered brownish outer region of the cuticle to dark/black. In contrast, RNAi of TmAANAT1 had no effect on the brown hue of the outer cuticle layer, but less or no pigmented inner region of the cuticle became significantly darker than those of control adults. These results suggest that, like that seen in TcAANAT1- or TcADC-deficient Tribolium castaneum adults, NADA produced by a reaction by TmAANAT1 contributes the lighter inner cuticle layer(s), whereas NBAD appears to do the highly pigmented outer cuticle layer(s) of the cuticle of T. molitor adults. This work was supported by NRFs (NRF-2015R1A6A3A04060323 and NRF-2018R1A2B6005106).

Insect structural cuticular proteins (CPs) play a major role in determining the diverse physical properties of the cuticle as a result of interactions/cross-linking among themselves and with chitin. CP genes compose a large gene family and have been classified more than ten distinct families based on the presence of unique amino acid sequence motifs. In this study, we performed RNAi-based functional analysis of eleven genes (TcCPLCP1-11) in Tribolium castaneum, which belong to CPLCP (Cuticular Proteins of Low Complexity, Proline rich) cuticular protein family. RNAi for TcCPLCP7-11 caused lethal pupal-adult molting defects and/or abnormal cuticle morphology in the resulting adults. Ultrastructural defects of the cuticles from TcCPLCP7-11-deficient insects by TEM are also discussed.

Insect cuticle/exoskeleton covering the entire external surface of the body is essential for protecting insects from various environmental stresses. Tyrosine metabolism plays a major role in not only the darkening of cuticle but also its hardening. In this work, we have focused on the functional analysis of nine genes involved in tyrosine-mediated cuticle tanning (pigmentation and sclerotization) pathway in Tenebrio molitor, which has a unique adult cuticle coloration, dark/black dorsal thorax and elytron, and reddish ventral thorax and abdomen. The temporal and spatial expression patterns of the genes were analyzed by real-time PCR, and RNA interference (RNAi) was performed to study the functional importance of these genes in cuticle coloration and/or hardening in T. molitor. This work was supported by NRFs (NRF-2015R1A2A2A01006614 and NRF-2015R1A6A3A04060323).

Chitin deacetylases (CDAs) are chitin-modifying enzymes that deacetylate chitin to form chitosan. In insects, this modification may contribute to the affinity and/or cross-linking of chitin/chitosan-like polysaccharides for a variety of structural proteins that may lead to diverse mechanical properties of the cuticle. DmCDA1 (serpentine) and DmCDA2 (vermiform) from Drosophila melanogaster play roles in development and morphology of embryonic tracheal tubes. We previously reported the functional importance of TcCDA1 and TcCDA2 orthologs to DmCDA1 and DmCDA2 from Tribolium castaneum, in molting, morphology of cuticle, and movement of legs. In this study, we further analyze ultrastructural defects of the cuticles and leg joints in TcCDA1- and TcCDA2-deficient insects. Loss of function of TcCDA1 and TcCDA2 causes disorganized chitinous horizontal laminae and vertical pore canals of rigid adult cuticle (e.g. elytron). Both proteins are also required for laminal organization in soft cuticle (e.g. hindwing). Morphological analysis of TcCDA1- and TcCDA2A-deficient adult revealed that ruptured tendons between femur and tibia cause the defects in movement of the leg joint. This work was supported by NRF (NRF-2015R1A2A2A01006614).

Since 2000s, proper utilization of IoT (Internet of Things) technology is a key factor for a firm to become more competitive, and this stream is not exceptional for the food and agriculture industry. Along with this stream, Korea government organization, for example MAFRA (Ministry of Agriculture, Food and Rural Affairs), elected to adopt IoT technology, such as USN and RFID technologies, in the food and agriculture industry. Based on the IoT technology, MAFARA launched six “IoT based farm” project in 2007. IoT based farm project includes IoT based greenhouse system project, and it shows drastic efficiency in terms of cost reduction. When it comes to the productivity, however, the effect of IoT based greenhouse system is still ambiguous. In this regard, this study conducted systems analysis and design for IoT based tomato greenhouse in order to help farmers’ decision making related to the productivity by establishing standardized database structure and designing output form to analyze productivity indices. Proposed systems analysis and design can be utilized as a data analysis tools by farmers. Productivity data from the proposed systems is can be used by researchers to identify the relationship among environment, plant growth and productivity. Policy makers also can refer to the data and output forms to predict the quantity of fruit during certain period and to revise production guideline more precisely.

This study aims to analyze the differences of ginseng product consumers and segment Korean fresh ginseng and red ginseng root markets based on attributes for the purchase. As a result of analyzing survey data, the red ginseng root consumers had different aspects from fresh ginseng consumers. According to the result of cluster analysis, the fresh ginseng consumers were subdivided into three segments (safety-oriented consumption cluster, label centered consumption cluster, and high involvement consumption cluster), while the red ginseng root consumers were subdivided into four segments (convenience-oriented consumption cluster, high involvement consumption cluster, raw material's safety-oriented cluster, and raw material's information importance cluster). ANOVA and Crosstab were conducted to investigate characteristics of each cluster.

In insect exoskeleton/cuticle, structural cuticular proteins (CPs) and the polysaccharide chitin are the major components of the procuticle. CPs are cross-linked by quinones or quinone methides produced by the laccase2 (Lac2)- mediated oxidation of N-acylcatechols. We reported that two major CPs, TcCPR27 and TcCPR18, belong to the CPR family that contain the RR-2 consensus motif (Rebers & Riddiford), are essential for formation and stabilization of the rigid cuticle of Tribolium castaneum adults. In this study, we characterized and investigated functions of the third most abundant protein, TcCP30, in extracts of elytra. TcCP30 cDNA encodes a protein with 171 amino acid residues containing a putative signal peptide. Unlike TcCPR27 and TcCPR18, TcCP30 mature protein lacks an RR motif, with a very unique amino composition, 36% Glu, 21% His, 20% Arg and 16% Gly. TcCP30 gene is highly expressed right before and after eclosion (in 5 d-old pupae and 0 d-old adults). Immunohistochemical studies revealed that TcCP30 protein was present in rigid cuticle such as elytra and ventral abdomen but not soft cuticle such as hindwings and dorsal abdomen of adult T. castaneum. Injection of dsRNA for TcCP30 into late instar larvae had no affect on larval and pupal growth and development. However, the subsequent pupal-adult molt, more than 50% adults were unable to shed their exuvium and died entrapped in their pupal cuticle. In addition, the resulting adults exhibited wrinkled, warped and split elytra. TcCP30-deficient adults could not fold their hindwings properly because probably due to the malformed elytra. These results indicate that TcCP30 is critical for formation of rigid adult cuticle as well as development and growth of T. castaneum.

Cuticular proteins (CPs) and the polysaccharide chitin are the major components of the exo- and endocuticular layers or procuticle. CPs contain a conserved sequence known as the Rebers & Riddiford (R&R) motif, which may function as a chitin-binding domain that helps to coordinate the interaction between chitin fibers and the protein network. We identified two highly abundant RR-2 CPs, TcCPR18 and TcCPR27, in protein samples extracted from elytra (rigid cuticle) of Tribolium castaneum adults and determined that these two CPs are required for rigid cuticle morphology. In this study, we identified the third most abundant protein (TcCP30) extracted from the elytra, and cloned a full-length cDNA. It encodes a very unusual 171 amino acid residue protein of which 36% of the residues of the mature protein are Glu, 21% are His, 19% are Arg, and 16% are Gly, organized in a regular pattern but not R&R consensus motif. TcCPR18 and TcCPR27 genes are expressed at 4 d-old pupae, while TcCP30 is highly expressed at 5 d-old pupae (last pupal stage) and 0 d-old adults. Immunohistochemical studies revealed the presence of TcCP30 in rigid adult cuticle (e.g. elytron, pronotum and ventral abdomen) but not soft cuticle (e.g. hindwing and dorsal abdomen). Injection of dsRNA for TcCP30 into late instar larvae had no affect on larval and pupal growth and development. The subsequent pupal-adult molt, however, more than 50% adults were unable to shed their exuvium and died. In addition, the resulting adults exhibited wrinkled, warped and split elytra. TcCP30-deficient adults could not fold their hindwings properly. These results indicate that TcCP30 may play critical roles in rigid adult cuticle formation, development and insect growth and survival. This work was supported by NRF (NRF-2012R1A2A1A01006467).