Jiuguang Wang

Molecular characterizations of natalisin and its roles in modulating mating in the oriental fruit fly, Bactrocera dorsalis (Hendel)

Initially, natalisin (NTL) was identified from three holometabolous insect species, Drosophila melanogaster, Tribolium castaneum and Bombyx mori, and was documented to regulate reproductive behaviours in D. melanogaster and T. castaneum. In this study, we report the sequences of the NTL precursor and its receptor (NTLR) from an important agricultural pest, Bactrocera dorsalis (Hendel). NTLR is a typical G‐protein coupled receptor and phylogenetic analysis showed that B. dorsalis NTLR was closely related to insect natalisin receptors from other species. A functional assay of NTLR transiently expressed in Chinese hamster ovary cells showed that it was activated by putative natalisin mature peptides in a concentration‐dependent manner, with 50% effective concentrations (EC50) at nanomolar or micromolar levels. As indicated by quantitative real‐time PCR, both NTL and NTLR had the highest expression in the central nervous system of B. dorsalis compared with the other tested tissues. Three pairs of adult brain neurones of B. dorsalis were identified with immunohistochemical antibody staining against D. melanogaster NTL4, and in situ hybridization with specific DNA probes. Moreover, RNA interference mediated by double‐stranded RNA injection in adults provided evidence for the important roles of NTL in regulating both male and female mating frequencies in this fly.

Identification and characterization of paternal-preferentially expressed gene NF - YC8 in maize endosperm

Gene imprinting describes an epigenetic phenomenon, whereby genetically identical alleles are differentially expressed dependent on parent-of-origin. Some imprinted genes belonged to NUCLEAR FACTOR Y (NF-Y) transcription factors, which were involved in many important metabolic processes in plant. The characterizations of imprinted genes are of great importance for their function exploration. In this paper, 15 non-redundant NF-YC genes were identified in the maize genome and the paternally expressed gene NF-YC8 was further analyzed. NF-YC8 primarily expressed in maize immature ear and tassel and phylogenetic analysis showed that NF-YC8 was highly homologous with Arabidopsis thalianaNF-YC2 genes which function in regulation of the flowering processes, ER stress response. Furthermore, NF-YC8 was a differential, gene-specific imprinted gene at 14 DAP and persistently imprinted throughout later endosperm development in the B73/Mo17 genetic background. Bisulfite sequencing for NF-YC8 in maize endosperm showed that the paternal alleles were higher methylated (CG, CHG and CHH contexts) than maternal alleles in the 5′ upstream region, and the coding region was highly methylated in CG context. Additionally, TE (CG, CHG and CHH contexts) and repetitive region (CG and CHG contexts) were all highly methylated. These results are the first description of evolution and molecular characterization of maize NF-YC8 and will provide new references for maize NF-YC genetic analysis.

Identification of QTL for acid phosphatase activity in root and rhizosphere soil of maize under low phosphorus stress

To provide theoretical and applied references for acid phosphatase (APase) activity of maize, this study was to identify quantitative trait locus (QTL) for APase activity in root and rhizosphere soil of maize under low phosphorus (p) stress. The correlation and the QTL of APase activity in root (APR) and APase activity in rhizosphere soil (APS) were studied for the F2:3 population derived from the cross 082×Ye107 under low P stress in two sites. Analysis for each environment and joint analysis across two environments were used to identify QTL for the F2:3 population. A significant difference in APR and APS was found between 082 (P efficient genotype) and Ye107 (P deficient genotype). A large genetic variation and transgressive segregation of the F2:3 population were observed in Beibei and Hechuan. One stable QTL for APR was detected in different environments, which was located in the interval bnlg1350 to bnlg1449 on chromosome 3. Two stable QTLs for APS were detected, which were located in the interval umc2083 to umc1972 on chromosome 1 and in the interval umc2111 to dupssr10 on chromosome 5. The stable QTLs can be used in MAS breeding and theoretical study of maize.

Vitellogenin and its receptor play essential roles in the development and reproduction of the brown citrus aphid, Aphis (Toxoptera) citricidus : Vg and VgR in brown citrus aphid

Vitellogenin (Vg) and its receptor (VgR) play a key role in the reproductive process and development of insects. Aphids are a group of high‐fecundity insect species with pseudoplacental viviparity, but the roles of their Vg and VgR genes have not been investigated yet. The brown citrus aphid, Aphis (Toxoptera) citricidus, is a major insect pest of citrus and the main vector of Citrus tristeza closterovirus. In this study, we identified and characterized these two genes, designated as AcVg and AcVgR, from the brown citrus aphid. We found that AcVg has lost the DUF1943 domain that is present in other insect Vgs. Silencing of AcVg and AcVgR led to a delay in the nymph–adult transition, a prolonged prereproductive period, and a shortened reproductive period, which in turn resulted in slower embryonic development and fewer new‐born nymphs. Interestingly, silencing of AcVg decreased the transcript level of AcVgR, but silencing of AcVgR resulted in increased transcript levels of AcVg. In addition, silencing of Vg/VgR had similar phenotypes between alate and apterous morphs, suggesting that the functions of these two genes are the same in the two wing morphs of the aphid. Our results demonstrate that Vg and VgR are involved in various aspects of aphid development and reproduction. Further studies on the synthesis of Vg could help to elucidate the reproductive mechanism and provide information that will be useful for developing new pest control strategies.

Overexpression of a maize plasma membrane intrinsic protein ZmPIP1;1 confers drought and salt tolerance in Arabidopsis

Drought and salt stress are major abiotic stress that inhibit plants growth and development, here we report a plasma membrane intrinsic protein ZmPIP1;1 from maize and identified its function in drought and salt tolerance in Arabidopsis. ZmPIP1;1 was localized to the plasma membrane and endoplasmic reticulum in maize protoplasts. Treatment with PEG or NaCl resulted in induced expression of ZmPIP1;1 in root and leaves. Constitutive overexpression of ZmPIP1;1 in transgenic Arabidopsis plants resulted in enhanced drought and salt stress tolerance compared to wild type. A number of stress responsive genes involved in cellular osmoprotection in ZmPIP1;1 overexpression plants were up-regulated under drought or salt condition. ZmPIP1;1 overexpression plants showed higher activities of reactive oxygen species (ROS) scavenging enzymes such as catalase and superoxide dismutase, lower contents of stress-induced ROS such as superoxide, hydrogen peroxide and malondialdehyde, and higher levels of proline under drought and salt stress than did wild type. ZmPIP1;1 may play a role in drought and salt stress tolerance by inducing of stress responsive genes and increasing of ROS scavenging enzymes activities, and could provide a valuable gene for further plant breeding.

Tudor knockdown disrupts ovary development in Bactrocera dorsalis : RNAi Tudor disrupts ovary development

One of the main functions of the piwi‐interacting RNA pathway is the post‐transcriptional silencing of transposable elements in the germline of many species. In insects, proteins belonging to the Tudor superfamily proteins belonging to the Tudor superfamily play an important role in to play an important role in this mechanism. In this study, we identified the tudor gene in the oriental fruit fly, Bactrocera dorsalis, investigated the spatiotemporal expressional profile of the gene, and performed a functional analysis using RNA interference. We identified one transcript for a tudor homologue in the B. dorsalis transcriptome, which encodes a protein containing the typical 10 Tudor domains and an Adenosine triphosphate (ATP) synthase delta subunit signature. Phylogenetic analysis confirmed the identity of this transcript as a tudor homologue in this species. The expression profile indicated a much higher expression in the adult and pupal stages compared to the larval stages (up to a 60‐fold increase), and that the gene was mostly expressed in the ovaries, Malpighian tubules and fat body. Finally, gene knockdown of tudor in B. dorsalis led to clearly underdeveloped ovaries in the female adult and reductions in copulation rate and amount of oviposition, indicating its important role in reproduction. The results of this study shed more light on the role of tudor in ovary development and reproduction.