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The ability of immature maize (Zea mays) embryos to form embryonic calluses (ECs) is highly genotype dependent, which limits transgenic breeding development in maize. Here, we report the association map-based cloning of ZmSAUR15 using an association panel (AP) consisting of 309 inbred lines with diverse formation abilities for ECs. We demonstrated that ZmSAUR15, which encodes a small auxin-upregulated RNA, acts as a negative effector in maize EC induction. Polymorphisms in the ZmSAUR15 promoter that influence the expression of ZmSAUR15 transcripts modulate the EC induction capacity in maize. ZmSAUR15 is involved in indole-3-acetic acid biosynthesis and cell division in immature embryo-derived callus. The ability of immature embryos to induce EC formation can be improved by the knockout of ZmSAUR15, which consequently increases the callus regeneration efficiency. Our study provides new insights into overcoming the genotypic limitations associated with EC formation and improving genetic transformation in maize.

Using GWAS and QTL mapping identified 100 QTL and 138 SNPs, which control yield-related traits in maize. The candidate gene GRMZM2G098557 was further validated to regulate ear row number by using a segregation population. Understanding the genetic basis of yield-related traits contributes to the improvement of grain yield in maize. This study used an inter-mated B73 × Mo17 (IBM) Syn10 doubled-haploid (DH) population and an association panel to identify the genetic loci responsible for nine yield-related traits in maize. Using quantitative trait loci (QTL) mapping, 100 QTL influencing these traits were detected across different environments in the IBM Syn10 DH population, with 25 co-detected in multiple environments. Using a genome-wide association study (GWAS), 138 single-nucleotide polymorphisms (SNPs) were identified as correlated with these traits (P

Two hub genes GRMZM2G075104 and GRMZM2G333183 involved in salt tolerance were identified by GWAS and WGCNA. Furthermore, they were verified to affect salt tolerance by candidate gene association analysis. Salt stress influences maize growth and development. To decode the genetic basis and hub genes controlling salt tolerance is a meaningful exploration for cultivating salt-tolerant maize varieties. Herein, we used an association panel consisting of 305 lines to identify the genetic loci responsible for Na+- and K+-related traits in maize seedlings. Under the salt stress, seven significant single nucleotide polymorphisms were identified using a genome-wide association study, and 120 genes were obtained by scanning the linkage disequilibrium regions of these loci. According to the transcriptome data of the above 120 genes under salinity treatment, we conducted a weighted gene co-expression network analysis. Combined the gene annotations, two SNaC/SKC (shoot Na+ content/shoot K+ content)-associated genes GRMZM2G075104 and GRMZM2G333183 were finally identified as the hub genes involved in salt tolerance. Subsequently, these two genes were verified to affect salt tolerance of maize seedlings by candidate gene association analysis. Haplotypes TTGTCCG-CT and CTT were determined as favorable/salt-tolerance haplotypes for GRMZM2G075104 and GRMZM2G333183, respectively. These findings provide novel insights into genetic architectures underlying maize salt tolerance and contribute to the cultivation of salt-tolerant varieties in maize.

There are still challenges for the development of multifunctional carbon nanotubes (CNTs). Here, a multiwalled carbon nanotube (MWCNT)-based rolling circle amplification system (CRCAS) is reported which allows in situ rolling circle replication of DNA primer on the surface of MWCNTs to create a long single-strand DNA (ssDNA) where a large number of nanoparticles or proteins could be loaded, forming a nano-biohybridized 3D structure with a powerful signal amplification ability. In this strategy, the binding ability of proteins, hybridization, replication ability of DNA, and the catalytical ability of enzymes are integrated on a single carbon nanotube. The CRCAS is then used to develop colorimetric and chemiluminescent assays for the highly sensitive and specific detection of cancer protein markers, alpha-fetoprotein (AFP) and prostate specific antigen (PSA). The colorimetric CRCAS assay is 4000 times more sensitive than a conventional enzyme-linked immunosorbent assay (ELISA), and its concentration range is 10 000 times wider. Control experiments show that as low as 10 pg mL(-1) AFP or PSA could be detected even in the presence of interfering protein markers with a more than 10(5)-fold greater concentration in the sample, demonstrating the high specificity of the CRCAS assay. The limit of detection of the chemiluminescent CRCAS assays for AFP and PSA are 5 fg mL(-1) (70 aM) and 10 fg mL(-1) (0.29 fM), respectively, indicating that the sensitivity is much higher than that of the colorimetric CRCAS assay. Importantly, CRCAS works well with real biological samples.

A portable and power-free microfluidic device was designed for rapid and sensitive detection of lead (Pb(2+)). 11-mercaptoundecanoic acid (MUA)-functionalized gold nanoparticles (MUA-AuNPs) aggregated in the presence of Pb(2+) for the chelation mechanism. When we performed this analysis on a polydimethylsiloxane (PDMS) microfluidic chip, the aggregations deposited onto the surface of chip and formed dark lines along the laminar flows in the zigzag microchannels. This visual result can be observed by the naked eye through a microscope or just a drop of water as a magnifier. Ten μM Pb(2+) was successfully detected.

Coupling nanomaterials with biomolecular recognition events represents a new direction in nanotechnology toward the development of novel molecular diagnostic tools. Here a graphene oxide (GO)-based multicolor fluorescent DNA nanoprobe that allows rapid, sensitive, and selective detection of DNA targets in homogeneous solution by exploiting interactions between GO and DNA molecules is reported. Because of the extraordinarily high quenching efficiency of GO, the fluorescent ssDNA probe exhibits minimal background fluorescence, while strong emission is observed when it forms a double helix with the specific targets, leading to a high signal-to-background ratio. Importantly, the large planar surface of GO allows simultaneous quenching of multiple DNA probes labeled with different dyes, leading to a multicolor sensor for the detection of multiple DNA targets in the same solution. It is also demonstrated that this GO-based sensing platform is suitable for the detection of a range of analytes when complemented with the use of functional DNA structures.

Biochemical and biomedical applications of graphene are critically dependent on the interaction between biomolecules and the nanomaterial. In this work, we developed a graphene-based signal-amplification nanoprobe by combining anti-immunoglobulin G (anti-IgG) and horseradish peroxidase (HRP) with graphene oxide (GO). The structure and function of HRP in the nano-interface of GO were firstly investigated, which demonstrated that the enzyme retained 78% of its native activity and 77% of its native a-helix content HRP and anti-IgG were then co-adsorbed onto GO to form bifunctional nanoprobes. The nanoprobes provide both improved binding ability and signal-amplification ability. Comparing with conventional bioconjugates such as enzyme-linked antibody, co-adsorption could avoid chemical conjugation between biomolecules, keeping their bioactivity well. As an example for their application, the nanoprobes were used to obtain amplified signals in a sandwich-type immunoassay for cancer marker, instead of conventional colorimetric conjugates. This approach provided a detection limit of 10 pg/mL alpha-fetoprotein (AFP), which was much more sensitive than conventional enzyme-linked immunosorbent assay (ELISA) methods. The easily fabricated GO-based nanoprobes have the potential to become universal probes for molecular diagnostics. (C) 2013 Elsevier B.V. All rights reserved.

Although “hot spots” have been proved to contribute to surface enhanced Raman scattering (SERS), less attention was paid to increase the number of the “hot spot” to directly enhance the Raman signals in bioanalytical systems. Here we report a new strategy based on nano rolling-circle amplification (nanoRCA) and nano hyperbranched rolling-circle amplification (nanoHRCA) to increase “hot spot” groups for protein microarrays. First, protein and ssDNA are coassembled on gold nanoparticles, making the assembled probe have both binding ability and hybridization ability. Second, the ssDNAs act as primers to initiate in situ RCA reaction to produced long ssDNAs. Third, a large number of SERS probes are loaded on the long ssDNA templetes, allowing thousands of SERS probes involved in each biomolecular recognition event. The strategy offered high-efficiency Raman enhancement and could detect less than 10 zeptomolar protein molecules in protein microarray analysis.

We have developed a surface-enhanced Raman scattering (SERS)-active substrate based on gold nanoparticle-decorated chemical vapor deposition (CVD)-growth graphene and used it for multiplexing detection of DNA. Due to the combination of gold nanoparticles and graphene, the Raman signals of dye were dramatically enhanced by this novel substrate. With the gold nanoparticles, DNA capture probes could be easily assembled on the surface of graphene films which have a drawback to directly immobilize DNA. This platform exhibits extraordinarily high sensitivity and excellent specificity for DNA detection. A detection limit as low as 10 pM is obtained. Importantly, two different DNA targets could be detected simultaneously on the same substrate just using one light source.

A fluorescence sensor for Ag(I) ions is developed based on the target-induced conformational change of a silver-specific cytosine-rich oligonucleotide (SSO) and the interactions between the fluorogenic SSO probe and graphene oxide.

Graphene is a particularly useful nanomaterial that has shown great promise in nanoelectronics. Because of the ultrahigh electron mobility of graphene and its unique surface properties such as one-atom thickness and irreversible protein adsorption at surfaces, graphene-based materials might serve as an ideal platform for accommodating proteins and facilitating protein electron transfer. In this work, we demonstrate that graphene oxide (GO) supports the efficient electrical wiring the redox centers of several heme-containing metalloproteins (cytochrome c, myoglobin, and horseradish peroxidase) to the electrode. Importantly, proteins retain their structural intactness and biological activity upon forming mixtures with GO. These important features imply the promising applications of GO/protein complexes in the development of biosensors and biofuel cells.

This paper proposes a simulation methodology to assess the risk in solder joint reliability (SJR) of Pin Grid Array (PGA)/ Land Grid Array (LGA) Socket during Mechanical shock. One of the main challenges in simulation is the complex socket/contact geometry convoluted with its massive number of degrees of freedom. This paper proposes a technique, Contact Isolation Method (CIM), to simplify the geometry so that it can be modeled with highly reduced degrees of freedom (DOFs) while still maintaining 90% of the in-plane and out-of-plane stiffness compared with the original structure. The Pin Grid Array 700 socket (PGA700) and Pin Grid Array 604 socket (PGA604) were used as case studies to demonstrate this methodology. A system model was completed for the PGA604 socket with a focus on its capability to assess SJR. A reasonable correlation between the modeling and experiment on the dynamic board strain demonstrates that this technique is viable. In addition, this paper verifies that the board strain is a proper metric for the risk assessment in SJR, and defines the optimal strain gauge location. The impact of the boundary conditions is also investigated.

This paper presents a numerical DOE to investigate the sensitivity of the mechanical performance of LGA contacts, e.g. contact working range, wiping distance and force, to four key design parameters: contact free height, contact thickness, contact material yield strength and the coefficient between the contact and its landing pad. Multiple cases were analyzed and the normalized response surfaces of the performance are presented. By keeping the linear terms of the response surfaces, the performance variances relative to the nominal can be estimated. The methodology using the estimated variances to define LGA contact design specification is also presented.Copyright © 2006 by ASME

We investigate interactions between graphene oxide and a Pb(2+)-dependent DNAzyme, based on which a Pb(2+) sensor with high sensitivity, selectivity and tunable dynamic range is developed.

A gold nanoprobe that can respond colorimetrically to Hg(2+) is designed and coupled with a power-free PDMS device; the system can be used for rapid and visual detection of low micromolar Hg(2+) in real environmental samples.