Expenditure Reform in Industrialised Countries: A Case Study Approach

This study examines reforms of public expenditure in industrialised countries over the past two decades. We distinguish ambitious and timid reformers and analyse in detail reform experiences in eight case studies of ambitious reform episodes. We find that ambitious reform countries reduce spending on transfers, subsidies and public consumption while largely sparing education spending. Such expenditure retrenchment is also typically part of a comprehensive reform package that includes improvements in fiscal institutions as well as structural and other macroeconomic reforms. The study finds that ambitious expenditure retrenchment and reform coincides with large improvements in fiscal and economic growth indicators

Characterization of Genetic Diversity and Linkage Disequilibrium of <em>ZmLOX4</em> and <em>ZmLOX5</em> Loci in Maize

<div><p>Maize (<em>Zea mays</em> L.) lipoxygenases (<em>ZmLOXs</em>) are well recognized as important players in plant defense against pathogens, especially in cross kingdom lipid communication with pathogenic fungi. This study is among the first to investigate genetic diversity at important gene paralogs <em>ZmLOX4</em> and <em>ZmLOX5</em>. Sequencing of these genes in 400 diverse maize lines showed little genetic diversity and low linkage disequilibrium in the two genes. Importantly, we identified one inbred line in which <em>ZmLOX5</em> has a disrupted open reading frame, a line missing <em>ZmLOX5</em>, and five lines with a duplication of <em>ZmLOX5</em>. Tajima's D test suggests that both <em>ZmLOX4</em> and <em>ZmLOX5</em> have been under neutral selection. Further investigation of haplotype data revealed that within the <em>ZmLOX</em> family members only <em>ZmLOX12</em>, a monocot specific <em>ZmLOX</em>, showed strong linkage disequilibrium that extends further than expected in maize. Linkage disequilibrium patterns at these loci of interest are crucial for future candidate gene association mapping studies. <em>ZmLOX4</em> and <em>ZmLOX5</em> mutations and copy number variants are under further investigation for crop improvement.</p> </div

Usefulness of Adapted Exotic Maize Lines Developed By Doubled Haploid and Single Seed Descent Methods

Adapted exotic maize (Zea mays L.) germplasm, such as BS39, provides a unique opportunity for broadening the genetic base of U.S. Corn Belt germplasm. In vivo doubled haploid (DH) technology has been used to efficiently exploit exotic germplasm. It can help to purge deleterious recessive alleles. The objectives of this study were to determine the usefulness of BS39-derived inbred lines using both SSD and DH methods, to determine the impact of spontaneous as compared to artificial haploid genome doubling on genetic variance among BS39-derived DH lines, and to identify SNP markers associated with agronomic traits among BS39 inbreds monitored at testcross level. We developed two sets of inbred lines directly from BS39 by DH and SSD methods, named BS39_DH and BS39_SSD. Additionally, two sets were derived from a cross between BS39 and A427 (SHGD donor) by DH and SSD methods, named BS39×A427_DH and BS39×A427_SSD, respectively. Grain yield, moisture, plant height, ear height, stalk lodging, and root lodging were measured to estimate genetic parameters. For genome-wide association (GWAS) analysis, inbred lines were genotyped using Genotype-by-Sequencing (GBS) and Diversity Array Technology Sequencing (DArTSeq). Some BS39-derived inbred lines performed better than elite germplasm inbreds and all sets showed significant genetic variance. The presence of spontaneous haploid genome doubling genes did not affect performance of inbred lines. Five SNPs were significant and three of them located within genes related to plant development or abiotic stresses. These results demonstrate the potential of BS39 to add novel alleles to temperate elite germplasm.This is a preprint made available through Research Square at doi:10.21203/rs.3.rs-799789/v1. This work is licensed under a CC BY 4.0 License

Sequence architecture of <i>ZmLOX5</i>.

<p><i>ZmLOX5</i> consists of 9 exons (shown in red), and 8 introns (shown in green). <i>ZmLOX5</i> has a shorter second intron, spanning 511 bp. <i>ZmLOX5</i> is located on the reverse strand of chromosome 5:12,274,159–12,279,067, spanning 4908 bp (B73 RefGen_V2). Vertical blue lines in the final 3′ exon of the gene are the relative locations of SNPs discovered from Sanger sequencing. An InDel of 28 bp was found in the inbred line Va99 and located in the final exon of <i>ZmLOX5</i>, shown here in light blue. Their base pair change, derived state (as compared to <i>Z. perennis</i>), and derived state percentages are shown in the grey boxes.</p

LD plot containing <i>ZmLOX12</i> and the 100 kb flanking the gene both upstream and downstream.

<p>The LD plot of SNPs from the Maize HapMap <i>ZmLOX12</i>, and the flaking 100 kb on either side of the gene shows that LD at this locus is very strong and extends farther than expected in maize. Approximately 3,000 bp across the <i>ZmLOX12</i> gene and extends beyond to approximately 19,000 bp. Further investigation of this locus revealed that there is a predicted gene of unknown function located very near (<100 bp) to <i>ZmLOX12</i>.</p

Sequence architecture of <i>ZmLOX4</i>.

<p>Similar to the <i>ZmLOX5</i> gene <i>ZmLOX4</i> consists of 9 exons (shown in red), and 8 introns (shown in green). The major difference between the architecture of the two genes is the length of the second intron, much larger in <i>ZmLOX4</i> spanning 11,191 bp (not shown). <i>ZmLOX4</i> is located on the forward strand of chromosome 1:264,209,651–264,226,078, spanning 16,427 bp (B73 RefGen_v2). Vertical blue lines in the final 3′ exon of the gene are the relative locations of SNPs discovered from Sanger sequencing. Their base pair change, derived state (as compared to <i>Z. perennis</i>), and derived state percentages are shown in the grey boxes.</p

LD plot containing 10 members of the <i>ZmLOX</i> gene family.

<p>The LD plot of all SNPs from the Maize HapMap with 10 members of the <i>ZmLOX</i> gene family (<i>ZmLOX2, ZmLOX4, ZmLOX5, ZmLOX6, ZmLOX7, ZmLOX8, ZmLOX9, ZmLOX10, ZmLOX11, and ZmLOX12</i>) ordered numerically from left to right shows little LD across all of the <i>ZmLOX</i>s except for <i>ZmLOX12</i>, which is located in the lower right-hand corner.</p

Southern blot of lines that did not PCR amplify for <i>ZmLOX5</i>.

<p>B73, confirmed to have a functioning ZmLOX5, is used as a control for the Southern blot. Five of the inbred lines screened show double banding: Yu796_NS, 4226, I29, HP301 and CI 187-2 and are suspected to have two copies of <i>ZmLOX5</i>. One is also missing a band: CML 247, and is suspected to have no copy of <i>ZmLOX5</i>.</p

LD patterns in the C-terminus exon of <i>ZmLOX4</i> (left) and <i>ZmLOX5</i> (right).

<p>LD plots of SNPs found via Sanger sequencing (outlined in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053973#pone-0053973-g001" target="_blank">Figures 1</a> and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053973#pone-0053973-g002" target="_blank">2</a>, and <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0053973#pone-0053973-t001" target="_blank">Table 1</a>) based on <i>ZmLOX4</i> with 6 SNPs and 1 InDel, spanning a total of 663 bp in the final 3′ exon across 260 diverse inbred lines, and <i>ZmLOX5</i> with 14 SNPs, spanning 709 bp across 203 diverse inbred lines. Both show a rapid (<100 bp) decay of LD (r²>0.1).</p

SNP locations per B73 RefGen_v2 and their derived state percentages.

<p>SNP locations per B73 RefGen_v2 and their derived state percentages.</p