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

A guiding role of the Arabidopsis circadian clock in cell differentiation revealed by time-series single-cell RNA sequencing

Circadian rhythms and progression of cell differentiation are closely coupled in multicellular organisms. However, whether establishment of circadian rhythms regulates cell differentiation or vice versa has not been elucidated due to technical limitations. Here, we exploit high cell fate plasticity of plant cells to perform single-cell RNA sequencing during the entire process of cell differentiation. By analyzing reconstructed actual time series of the differentiation processes at single-cell resolution using a method we developed (PeakMatch), we find that the expression profile of clock genes is changed prior to cell differentiation, including induction of the clock gene LUX ARRYTHMO (LUX). ChIP sequencing analysis reveals that LUX induction in early differentiating cells directly targets genes involved in cell-cycle progression to regulate cell differentiation. Taken together, these results not only reveal a guiding role of the plant circadian clock in cell differentiation but also provide an approach for time-series analysis at single-cell resolution

Dense Clumps in Giant Molecular Clouds in the Large Magellanic Cloud: Density and Temperature Derived from 13^{13}CO(J=3−2J=3-2) Observations

In order to precisely determine temperature and density of molecular gas in the Large Magellanic Cloud, we made observations of optically thin $^{13}$CO($J=3-2$) transition by using the ASTE 10m telescope toward 9 peaks where $^{12}$CO($J=3-2$) clumps were previously detected with the same telescope. The molecular clumps include those in giant molecular cloud (GMC) Types I (with no signs of massive star formation), II (with HII regions only), and III (with HII regions and young star clusters). We detected $^{13}$CO($J=3-2$) emission toward all the peaks and found that their intensities are 3 -- 12 times lower than those of $^{12}$CO($J=3-2$). We determined the intensity ratios of $^{12}$CO($J=3-2$) to $^{13}$CO($J=3-2$), $R^{12/13}_{3-2}$, and $^{13}$CO($J=3-2$) to $^{13}$CO($J=1-0$), $R^{13}_{3-2/1-0}$, at 45\arcsec resolution. These ratios were used for radiative transfer calculations in order to estimate temperature and density of the clumps. The parameters of these clumps range kinetic temperature $T\mathrm{_{kin}}$ = 15 -- 200 K, and molecular hydrogen gas density $n(\mathrm{H_2})$ = 8$\times 10^2$ -- 7$\times 10^3$ cm$^{-3}$. We confirmed that the higher density clumps show higher kinetic temperature and that the lower density clumps lower kinetic temperature at a better accuracy than in the previous work. The kinetic temperature and density increase generally from a Type I GMC to a Type III GMC. We interpret that this difference reflects an evolutionary trend of star formation in molecular clumps. The $R^{13}_{3-2/1-0}$ and kinetic temperature of the clumps are well correlated with H$\alpha$ flux, suggesting that the heating of molecular gas $n(\mathrm{H_2})$ = $10^3$ -- $10^4$ cm$^{-3}$ can be explained by stellar FUV photons.Comment: 39 pages, 7 figures, 4 tables. Accepted for publication in The Astronomical Journa

Decentralized circadian clocks process thermal and photoperiodic cues in specific tissues

植物の体内時計が日の長さと温度の情報を異なる組織で処理していることを発見. 京都大学プレスリリース. 2015-11-04.The circadian clock increases organisms' fitness by regulating physiological responses1. In mammals, the circadian clock in the suprachiasmatic nucleus (SCN) governs daily behavioural rhythms2. Similarly, in Arabidopsis, tissue-specific circadian clock functions have emerged, and the importance of the vasculature clock for photoperiodic flowering has been demonstrated. However, it remains unclear if the vasculature clock regulates the majority of physiological responses, like the SCN in mammals, and if other environmental signals are also processed by the vasculature clock. Here, we studied the involvement of tissue-specific circadian clock regulation of flowering and cell elongation under different photoperiods and temperatures. We found that the circadian clock in vascular phloem companion cells is essential for photoperiodic flowering regulation; by contrast, the epidermis has a crucial impact on ambient temperature-dependent cell elongation. Thus, there are clear assignments of roles among circadian clocks in each tissue. Our results reveal that, unlike the more centralized circadian clock in mammals, the plant circadian clock is decentralized, where each tissue specifically processes individual environmental cues and regulates individual physiological responses. Our new conceptual framework will be a starting point for deciphering circadian clock functions in each tissue, which will lead to a better understanding of how circadian clock processing of environmental signals may be affected by ongoing climate change