A new containerless image furnace with electro-static positioning device

A new containerless image furnace with a microwave discharge plasma lamp and electrostatic positioning device was developed for the use of the microgravity experiment on the Japanese experimental module (JEM). The electrostatic positioning system was tested under the reduced gravity environment in the MU-300 aircraft. Solid specimens (maximum weight is 1.3 gr and 10 mm in diameter) and water drops (maximum weight is 0.11 gr and 6 mm in diameter) were successfully controlled under the 0.02G environment. Rotation control of the dielectric specimen was also possible by means of supplying a rotating electric field while the specimen is levitating. The measured rotation speed of the glass shell specimen (0.08 gr, 10 mm) was up to 110 rpm, when the rotating field frequency was 6Hz

Multiplex Genome Editing in Tomato

Several expression systems for multiple guide RNA (gRNAs) have been developed in the CRISPR/Cas9 (clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9) system to induce multiple-gene modifications in plants. Here, we evaluated mutation efficiencies in the tomato genome using multiplex CRISPR/Cas9 vectors consisting of various Cas9 expression promoters with multiple gRNA expression combinations. In transgenic tomato calli induced with these vectors, mutation patterns varied depending on the promoters used to express Cas9. By using the tomato ELONGATION FACTOR-1α (SlEF1α) promoter to drive Cas9, occurrence of various types of mutations with high efficiency was detected in the tomato genome. Furthermore, sequence analysis showed that the majority of mutations using the multiplex system with the SlEF1α promoter corresponded to specific mutation pattern of deletions produced by self-ligation at two target sites of CRISPR/Cas9 with low mosaic mutations. These results suggest that optimizing the Cas9 expression promoter used in CRISPR/Cas9-mediated mutation improves multiplex genome editing, and could be used effectively to disrupt functional domains precisely in the tomato genome

SlIAA9 Controls Tomato Elongation

Tomato INDOLE-3-ACETIC ACID9 (SlIAA9) is a transcriptional repressor in auxin signal transduction, and SlIAA9 knockout tomato plants develop parthenocarpic fruits without fertilization. We generated sliaa9 mutants with parthenocarpy in several commercial tomato cultivars (Moneymaker, Rio Grande, and Ailsa Craig) using CRISPR-Cas9, and null-segregant lines in the T1 generation were isolated by self-pollination, which was confirmed by PCR and Southern blot analysis. We then estimated shoot growth phenotypes of the mutant plants under different light (low and normal) conditions. The shoot length of sliaa9 plants in Moneymaker and Rio Grande was smaller than those of wild-type cultivars in low light conditions, whereas there was not clear difference between the mutant of Ailsa Craig and the wild-type under both light conditions. Furthermore, young seedlings in Rio Grande exhibited shade avoidance response in hypocotyl growth, in which the hypocotyl lengths were increased in low light conditions, and sliaa9 mutant seedlings of Ailsa Craig exhibited enhanced responses in this phenotype. Fruit production and growth rates were similar among the sliaa9 mutant tomato cultivars. These results suggest that control mechanisms involved in the interaction of AUX/IAA9 and lights condition in elongation growth differ among commercial tomato cultivars

Narrowing Down the Mapping of Plant Sex-Determination Regions Using New Y-Chromosome-Specific Markers and Heavy-Ion Beam Irradiation-Induced Y-Deletion Mutants in Silene latifolia

Silene latifolia is a well-studied model system for plant XY sex determination. Three maleness factors are thought to function on the Y chromosome, gynoecium suppression factor (GSF), stamen-promoting factor (SPF), and male fertility factor (MFF), and their deletions result in hermaphrodites, anther defects, and pollen defects, respectively. Although a framework map of the Y chromosome exists, the sex determination genes have not been identified, and no markers close enough to potentially be used for BAC library screening are yet available. The analysis of Y deletion mutants by Y-chromosome-specific STS markers is an efficient way to isolate sex determination regions, but more Y-specific STS markers are needed to accelerate the exploration of sex determination factors. Herein, we report a marker design method that uses simple sequence repeats, which is especially effective on the Y chromosome of S. latifolia because it contains many simple sequence repeats. Six new Y-chromosome-specific STS markers were obtained, SmicSy1–6. These were used to detect relatively small Y deletion sites in heavy-ion beam irradiation-induced mutants. The mapping of male sex determination regions was narrowed down by using more markers and smaller-sized Y deletion mutants. One new marker, SmicSy6, is a proximal marker to SPF and, thus, a second index for SPF. The region including SPF is thought to be located between two SPF proximal markers. The flower phenotype correlates with the deletion size of SPF using SPF proximal markers. These findings represent new progress in isolating the sex determination factor, which has been studied for more than 50 years

Genome editing in plants using CRISPR type I-D nuclease

Genome editing in plants has advanced greatly by applying the clustered regularly interspaced short palindromic repeats (CRISPRs)-Cas system, especially CRISPR-Cas9. However, CRISPR type I—the most abundant CRISPR system in bacteria—has not been exploited for plant genome modification. In type I CRISPR-Cas systems, e.g., type I-E, Cas3 nucleases degrade the target DNA in mammals. Here, we present a type I-D (TiD) CRISPR-Cas genome editing system in plants. TiD lacks the Cas3 nuclease domain; instead, Cas10d is the functional nuclease in vivo. TiD was active in targeted mutagenesis of tomato genomic DNA. The mutations generated by TiD differed from those of CRISPR/Cas9; both bi-directional long-range deletions and short indels mutations were detected in tomato cells. Furthermore, TiD can be used to efficiently generate bi-allelic mutant plants in the first generation. These findings indicate that TiD is a unique CRISPR system that can be used for genome engineering in plants

Do abstraction and power increase moral hypocrisy?

Lammers (2012) reported that participants taking an abstract (vs. concrete) view tended to judge the immoral behavior of others more severely than the same behavior performed by themselves (i.e., moral hypocrisy). In the current study, we examined the effects of construal level (i.e., abstract vs. concrete mindset) on moral judgments in a sample of Japanese university students. In addition, we examined the effect of the individuals’ sense of power, which was previously found to influence moral hypocrisy (Lammers et al., 2010); people in powerful roles were found to be more strict in moral judgments of others’ behavior than their own behavior. However, in Experiment 1, we did not observe moral hypocrisy. Instead, regardless of the actors in scenarios, we found that participants in the abstract condition (vs. concrete condition) judged immoral behavior less severely. Moreover, participants with a greater sense of power were more likely to judge immoral behavior severely. The discrepancy between the current findings and those of previous studies might be related to the scenario used in the current study (stealing a bike in a public bicycle parking area). Participants may have been influenced by the concern of being watched by a third party. We conducted Experiment 2 to investigate the effects of the presence of a third party, but found no significant effects. The results are discussed in terms of the construal level manipulations and immoral behavior in the experimental scenarios.本論文は，2017年度に広島大学教育学部で開講された心理学課題演習において，第1著者の指導により第2著者から第7著者までが実施した研究をもとに執筆したものである。研究の一部は第2著者から第7著者により中国四国心理学会第73回大会学部生研究発表会において報告された