The PIWI‐Interacting RNA Molecular Pathway: Insights From Cultured Silkworm Germline Cells

The PIWI‐interacting RNA (piRNA) pathway, one of the major eukaryotic small RNA silencing pathways, is a genome surveillance system that silences selfish genes in animal gonads. piRNAs guide PIWI protein to target genes through Watson–Crick RNA–RNA base‐parings. Loss of piRNA function causes genome instability, inducing failure in gametogenesis and infertility. Studies using fruit flies and mice as key experimental models have resulted in tremendous progress in understanding the mechanism underlying the piRNA pathway. Recent work using cultured silkworm germline cells has also expanded our knowledge of piRNA biogenesis in particular, since these silkworm cells are the only cells of germline origin that can be cultured. In this review, we describe elucidation of the piRNA pathway using cultured silkworm cells as an experimental model by focusing on recent work in biochemistry and structural biology. Earlier studies that made important contributions to the field are also described.     

Breeding for salt tolerance in crop plants — the role of molecular biology

Salinity in soil affects about 7 % of the land’s surface and about 5 % of cultivated land. Most importantly, about 20 % of irrigated land has suffered from secondary salinisation and 50 % of irrigation schemes are affected by salts. In many hotter, drier countries of the world salinity is a concern in their agriculture and could become a key issue. Consequently, the development of salt resistant crops is seen as an important area of research. Although there has been considerable research into the effects of salts on crop plants, there has not, unfortunately, been a commensurate release of salt tolerant cultivars of crop plants. The reason is likely to be the complex nature of the effect of salts on plants. Given the rapid increase in molecular biological techniques, a key question is whether such techniques can aid the development of salt resistance in plants. Physiological and biochemical research has shown that salt tolerance depends on a range of adaptations embracing many aspects of a plant’s physiology: one of these the compartmentation of ions. Introducing genes for compatible solutes, a key part of ion compartmentation, in salt-sensitive species is, conceptually, a simple way of enhancing tolerance. However, analysis of the few data available suggests the consequences of transformation are not straightforward. This is not unexpected for a multigenic trait where the hierarchy of various aspects of tolerance may differ between and within species. The experimental evaluation of the response of transgenic plants to stress does not always match, in quality, the molecular biology. We have advocated the use of physiological traits in breeding programmes as a process that can be undertaken at the present while more knowledge of the genetic basis of salt tolerance is obtained. The use of molecular biological techniques might aid plant breeders through the development of marker aided selection.     

Gene expression and localization of a β-1,3-glucanase of <Emphasis Type="Italic">Lotus japonicus</Emphasis>

Phytohormone abscisic acid (ABA) inhibits root nodule formation of leguminous plants. LjGlu1, a β-1,3-glucanase gene of Lotus japonicus, has been identified as an ABA responsive gene. RNA interference of LjGlu1 increased nodule number. This suggests that LjGlu1 is involved in the regulation of nodule formation. Host legumes control nodule number by autoregulation of nodulation (AON), in which the presence of existing root nodules inhibits further nodulation. For further characterization of LjGlu1, we focused on the expression of LjGlu1 in relation to AON. In a split-root system, LjGlu1 expression peaked when AON was fully induced. Hairy roots transformed with LjCLE-RS1, a gene that induces AON, were generated. Expression of LjGlu1 was greater in the transgenic roots than in untransformed roots. LjGlu1 was not induced in a hypernodulating mutant inoculated with Mesorhizobium loti. These results suggest that the expression of LjGlu1 is involved in the system of AON. However, neither hypernodulation nor enlarged nodulation zone was observed on the transgenic hairy roots carrying LjGlu1-RNAi, suggesting that LjGlu1 is not a key player of AON. Recombinant LjGlu1 showed endo-β-1,3-glucanase activity. LjGlu1-mOrange fusion protein suggested that LjGlu1 associated with M. loti on the root hairs. Exogenous β-1,3-glucanase inhibited infection thread formation by both the wild type and the mutant, and nodule numbers were reduced. These results suggest that LjGlu1 is expressed in response to M. loti infection and functions outside root tissues, resulting in the inhibition of infection.     

Development and evaluation of an electromagnetic hypersensitivity questionnaire for Japanese people

The purpose of the present study was to evaluate the validity and reliability of a Japanese version of an electromagnetic hypersensitivity (EHS) questionnaire, originally developed by Eltiti et al. in the United Kingdom. Using this Japanese EHS questionnaire, surveys were conducted on 1306 controls and 127 self‐selected EHS subjects in Japan. Principal component analysis of controls revealed eight principal symptom groups, namely, nervous, skin‐related, head‐related, auditory and vestibular, musculoskeletal, allergy‐related, sensory, and heart/chest‐related. The reliability of the Japanese EHS questionnaire was confirmed by high to moderate intraclass correlation coefficients in a test–retest analysis, and high Cronbach's α coefficients (0.853–0.953) from each subscale. A comparison of scores of each subscale between self‐selected EHS subjects and age‐ and sex‐matched controls using bivariate logistic regression analysis, Mann–Whitney U‐ and χ² tests, verified the validity of the questionnaire. This study demonstrated that the Japanese EHS questionnaire is reliable and valid, and can be used for surveillance of EHS individuals in Japan. Furthermore, based on multiple logistic regression and receiver operating characteristic analyses, we propose specific preliminary criteria for screening EHS individuals in Japan. Bioelectromagnetics. 37:353–372, 2016. © 2016 The Authors. Bioelectromagnetics Published by Wiley Periodicals, Inc.     

Cytokinin Activity of cis-Zeatin and Phenotypic Alterations Induced by Overexpression of Putative cis-Zeatin-O-glucosyltransferase in Rice

cis-Zeatin (cZ) is generally regarded as a cytokinin with little or no activity, compared with the highly active trans-zeatin (tZ). Although recent studies suggested possible roles for cZ, its physiological significance remains unclear. In our studies with rice (Oryza sativa), cZ inhibited seminal root elongation and up-regulated cytokinin-inducible genes, and its activities were comparable to those of tZ. Tracer experiments showed that exogenously supplied cZ-riboside was mainly converted into cZ derivatives but scarcely into tZ derivatives, indicating that isomerizations of cZ derivatives into tZ derivatives are a minor pathway in rice cytokinin metabolism. We identified three putative cZ-O-glucosyltransferases (cZOGT1, cZOGT2, and cZOGT3) in rice. The cZOGTs preferentially catalyzed O-glucosylation of cZ and cZ-riboside rather than tZ and tZ-riboside in vitro. Transgenic rice lines ectopically overexpressing the cZOGT1 and cZOGT2 genes exhibited short-shoot phenotypes, delay of leaf senescence, and decrease in crown root number, while cZOGT3 overexpressor lines did not show shortened shoots. These results propose that cZ activity has a physiological impact on the growth and development of rice.