Nuclear Transcriptomes at High Resolution Using Retooled INTACT.

Isolated nuclei provide access to early steps in gene regulation involving chromatin as well as transcript production and processing. Here, we describe transfer of the isolation of nuclei from tagged specific cell types (INTACT) to the monocot rice (Oryza sativa L.). The purification of biotinylated nuclei was redesigned by replacing the outer nuclear-envelope-targeting domain of the nuclear tagging fusion (NTF) protein with an outer nuclear-envelope-anchored domain. This modified NTF was combined with codon-optimized Escherichia coli BirA in a single T-DNA construct. We also developed inexpensive methods for INTACT, T-DNA insertion mapping, and profiling of the complete nuclear transcriptome, including a ribosomal RNA degradation procedure that minimizes pre-ribosomal RNA (pre-rRNA) transcripts. A high-resolution comparison of nuclear and steady-state poly(A)+ transcript populations of seedling root tips confirmed the capture of pre-messenger RNA (pre-mRNA) and exposed distinctions in diversity and abundance of the nuclear and total transcriptomes. This retooled INTACT can enable high-resolution monitoring of the nuclear transcriptome and chromatin in specific cell types of rice and other species.

Axiomatic opportunities and obstacles for inferring a species tree from gene trees.

The reconstruction of a central tendency "species tree" from a large number of conflicting gene trees is a central problem in systematic biology. Moreover, it becomes particularly problematic when taxon coverage is patchy, so that not all taxa are present in every gene tree. Here, we list four apparently desirable properties that a method for estimating a species tree from gene trees could have (the strongest property states that building a species tree from input gene trees and then pruning leaves gives a tree that is the same as, or more resolved than, the tree obtained by first removing the taxa from the input trees and then building the species tree). We show that although it is technically possible to simultaneously satisfy these properties when taxon coverage is complete, they cannot all be satisfied in the more general supertree setting. In part two, we discuss a concordance-based consensus method based on Baum's "plurality clusters", and an extension to concordance supertrees.

Gene regulatory networks shape developmental plasticity of root cell types under water extremes in rice.

Understanding how roots modulate development under varied irrigation or rainfall is crucial for development of climate-resilient crops. We established a toolbox of tagged rice lines to profile translating mRNAs and chromatin accessibility within specific cell populations. We used these to study roots in a range of environments: plates in the lab, controlled greenhouse stress and recovery conditions, and outdoors in a paddy. Integration of chromatin and mRNA data resolves regulatory networks of the following: cycle genes in proliferating cells that attenuate DNA synthesis under submergence; genes involved in auxin signaling, the circadian clock, and small RNA regulation in ground tissue; and suberin biosynthesis, iron transporters, and nitrogen assimilation in endodermal/exodermal cells modulated with water availability. By applying a systems approach, we identify known and candidate driver transcription factors of water-deficit responses and xylem development plasticity. Collectively, this resource will facilitate genetic improvements in root systems for optimal climate resilience.

What Has Been the Focus of Sugarcane Research? A Bibliometric Overview.

Sugarcane is one of the main crops worldwide, and it has an important impact on environmental issues. A bibliometric mapping analysis of the research on sugarcane was carried out, using data on the titles, abstracts, and keywords of articles published in leading journals and other peer-reviewed documents available in the SCOPUS database from 1858 to 2019 (27 August), and this was subsequently analyzed with the software VOSviewer. The three most important countries that publish research and were most-cited regarding sugarcane were Brazil, the USA, and India. The analysis of the co-occurrence of terms shows that the main research areas were sugarcane bagasse and terms related to bioenergy, and on a second level of relevance agronomy topics related to increasing crop yields. This first attempt to visualize the abundance of publications regarding sugarcane in their totality is in itself a good starting point for further scientific discussion.

Species concepts should not conflict with evolutionary history, but often do.

Many phylogenetic systematists have criticized the Biological Species Concept (BSC) because it distorts evolutionary history. While defences against this particular criticism have been attempted, I argue that these responses are unsuccessful. In addition, I argue that the source of this problem leads to previously unappreciated, and deeper, fatal objections. These objections to the BSC also straightforwardly apply to other species concepts that are not defined by genealogical history. What is missing from many previous discussions is the fact that the Tree of Life, which represents phylogenetic history, is independent of our choice of species concept. Some species concepts are consistent with species having unique positions on the Tree while others, including the BSC, are not. Since representing history is of primary importance in evolutionary biology, these problems lead to the conclusion that the BSC, along with many other species concepts, are unacceptable. If species are to be taxa used in phylogenetic inferences, we need a history-based species concept.

Isolation of Nuclei in Tagged Cell Types (INTACT), RNA Extraction and Ribosomal RNA Degradation to Prepare Material for RNA-Seq.

Gene expression is dynamically regulated on many levels, including chromatin accessibility and transcription. In order to study these nuclear regulatory events, we describe our method to purify nuclei with Isolation of Nuclei in TAgged Cell Types (INTACT). As nuclear RNA is low in polyadenylated transcripts and conventional pulldown methods would not capture non-polyadenylated pre-mRNA, we also present our method to remove ribosomal RNA from the total nuclear RNA in preparation for nuclear RNA-Seq.

Objective and subjective probability in gene expression.

In this paper I address the question of whether the probabilities that appear in models of stochastic gene expression are objective or subjective. I argue that while our best models of the phenomena in question are stochastic models, this fact should not lead us to automatically assume that the processes are inherently stochastic. After distinguishing between models and reality, I give a brief introduction to the philosophical problem of the interpretation of probability statements. I argue that the objective vs. subjective distinction is a false dichotomy and is an unhelpful distinction in this case. Instead, the probabilities in our models of gene expression exhibit standard features of both objectivity and subjectivity.

Ammonia emissions during vermicomposting of sheep manure.

The effect of C:N ratio, temperature and water content on ammonia volatilization during two-phase composting of sheep manure was evaluated. The aerobic phase was conducted under field conditions. This was followed by Phase II, vermicomposting, conducted in the laboratory under controlled conditions of water content (70% and 80%) and temperature (15 and 22 °C). The addition of extra straw lead to a 10% reduction in NH3 volatilization compared to sheep manure composted without extra straw. Temperature and water content significantly effected ammonia volatilization at 0 day in Phase II, with a water content of 70% and temperature of 22 °C leading to greater losses of ammonia. Nitrogen loss by ammonia volatilization during vermicomposting ranged from 8% to 15% of the initial N content. The addition of extra straw did not result in significant differences in total carbon content following vermicomposting.