In this paper, the development of the Paphiopedilum Maudiae embryo sac at different developmental stages after pollination was assessed by confocal laser scanning microscopy. The mature seeds of P. Maudiae consisted of an exopleura and a spherical embryo, but without an endosperm, while the inner integument cells were absorbed by the developing embryo. The P. Maudiae embryo sac exhibited an Allium type of development. The time taken for the embryo to develop to a mature sac was 45-50 days after pollination (DAP) and most mature embryo sacs had completed fertilization and formed zygotes by about 50–54 DAP. In planta transformation was achieved by injection of the ovaries by Agrobacterium, resulting in 38 protocorms or seedlings after several rounds of hygromycin selection, corresponding to 2, 7, 5, 1, 3, 4, 9, and 7 plantlets from Agrobacterium-mediated ovary-injection at 30, 35, 42, 43, 45, 48, 50, and 53 DAP, respectively. Transformation efficiency was highest at 50 DAP (2.54%), followed by 2.48% at 53 DAP and 2.45% at 48 DAP. Four randomly selected hygromycin-resistant plants were GUS-positive after PCR analysis. Semi-quantitative PCR and quantitative real-time PCR analysis revealed the expression of the hpt gene in the leaves of eight hygromycin-resistant seedlings following Agrobacterium-mediated ovary-injection at 30, 35, 42, 43, 45, 48, 50, and 53 DAP, while hpt expression was not detected in the control. The best time to inject P. Maudiae ovaries in planta with Agrobacterium is 48-53 DAP, which corresponds to the period of fertilization. This protocol represents the first genetic transformation protocol for any Paphiopedilum species and will allow for expanded molecular breeding programs to introduce useful and interesting genes that can expand its ornamental and horticulturally important characteristics. 相似文献
Journal of Porous Materials - The dynamics and thermodynamics of adsorption of hexadecyl ammonium with different numbers of carbon chains in montmorillonite (Mt) with different layer charge density... 相似文献
Neo-tetraploid rice with high fertility is a useful germplasm for polyploid rice breeding, which was developed from the crossing of different autotetraploid rice lines. However, little information is available on the molecular mechanism underlying the fertility of neo-tetraploid rice. Here, two contrasting populations of tetraploid rice, including one with high fertility (hereafter referred to as JG) and another with low fertility (hereafter referred to as JD), were generated by crossing Huaduo 3 (H3), a high fertility neo-tetraploid rice that was developed by crossing Jackson-4x with 96025-4x, and Huajingxian74-4x (T452), a low fertility autotetraploid rice parent. Cytological, global genome sequencing-based bulked-segregant (BSA-seq) and CRISPR/Cas9 technology were employed to study the genes associated with pollen fertility in neo-tetraploid rice. The embryo sacs of JG and JD lines were normal; however, pollen fertility was low in JD, which led to scarce fertilization and low seed setting. Cytological observations displayed low pollen fertility (25.1%) and approximately 31.3 and 27.2% chromosome lagging at metaphase I and II, and 28.8 and 24.8% chromosome straggling at anaphase I and II in JD, respectively. BSA-seq of F2–3 generations and RNA-seq of F4 generation detected a common fragment, i.e., 18,915,234–19,500,000, at chromosome 7, which was comprised of 78 genes associated with fertility. Among 78 genes, 9 genes had been known to be involved in meiosis and pollen development. Two mutants ny1 (LOC_Os07g32406) and ny2 (LOC_Os07g32040) were generated by CRISPR/Cas9 knockout in neo-tetraploid rice, and which exhibited low pollen fertility and abnormal chromosome behavior. Our study revealed that two unknown genes, LOC_Os07g32406 (NY1) and LOC_Os07g32040 (NY2) play an important role in pollen development of neo-tetraploid rice and provides a new perspective about the genetic mechanisms of fertility in polyploid rice. 相似文献
Sphingosine-1-phosphate (S1P), is a signaling sphingolipid which acts as a bioactive lipid mediator. We assessed whether S1P had multiplex effects in regulating the large-conductance Ca2+-activated K+ channel (BKCa) in catecholamine-secreting chromaffin cells. Using multiple patch-clamp modes, Ca2+ imaging, and computational modeling, we evaluated the effects of S1P on the Ca2+-activated K+ currents (IK(Ca)) in bovine adrenal chromaffin cells and in a pheochromocytoma cell line (PC12). In outside-out patches, the open probability of BKCa channel was reduced with a mean-closed time increment, but without a conductance change in response to a low-concentration S1P (1 µM). The intracellular Ca2+ concentration (Cai) was elevated in response to a high-dose (10 µM) but not low-dose of S1P. The single-channel activity of BKCa was also enhanced by S1P (10 µM) in the cell-attached recording of chromaffin cells. In the whole-cell voltage-clamp, a low-dose S1P (1 µM) suppressed IK(Ca), whereas a high-dose S1P (10 µM) produced a biphasic response in the amplitude of IK(Ca), i.e., an initial decrease followed by a sustained increase. The S1P-induced IK(Ca) enhancement was abolished by BAPTA. Current-clamp studies showed that S1P (1 µM) increased the action potential (AP) firing. Simulation data revealed that the decreased BKCa conductance leads to increased AP firings in a modeling chromaffin cell. Over a similar dosage range, S1P (1 µM) inhibited IK(Ca) and the permissive role of S1P on the BKCa activity was also effectively observed in the PC12 cell system. The S1P-mediated IK(Ca) stimulation may result from the elevated Cai, whereas the inhibition of BKCa activity by S1P appears to be direct. By the differentiated tailoring BKCa channel function, S1P can modulate stimulus-secretion coupling in chromaffin cells. 相似文献
With the continuous expansion of power systems and the application of power electronic equipment, forced oscillation has become one of the key problems in terms of system safety and stability. In this paper, an interline power flow controller (IPFC) is used as a power suppression carrier and its mechanism is analyzed using the linearized state-space method to improve the system damping ratio. It is shown that although the IPFC can suppress forced oscillation with well-designed parameters, its capability of improving the system damping ratio is limited. Thus, combined with the repetitive control method, an additional repetitive controller (ARC) is proposed to further dampen the forced power oscillation. The ARC control scheme is characterized by outstanding tracking performance to a system steady reference value, and the main IPFC controller with the ARC can provide higher damping, and further reduce the amplitude of oscillations to zero compared with a supplementary damping controller (SDC). Simulation results show that the IPFC with an ARC can not only greatly reduce the oscillation amplitude, but also actively output the compensation power according to the reference value of the ARC tracking system. 相似文献
Efficiently performing high-resolution direction of arrival (DOA) estimation under low signal-to-noise ratio (SNR) conditions has always been a challenge task in the literatures. Obvi-ously, in order to address this problem, the key is how to mine or reveal as much DOA related in-formation as possible from the degraded array outputs. However, it is certain that there is no per-fect solution for low SNR DOA estimation designed in the way of winner-takes-all. Therefore, this paper proposes to explore in depth the complementary DOA related information that exists in spa-tial spectrums acquired by different basic DOA estimators. Specifically, these basic spatial spec-trums are employed as the input of multi-source information fusion model. And the multi-source in-formation fusion model is composed of three heterogeneous meta learning machines, namely neural networks (NN), support vector machine (SVM), and random forests (RF). The final meta-spec-trum can be obtained by performing a final decision-making method. Experimental results illus-trate that the proposed information fusion based DOA estimation method can really make full use of the complementary information in the spatial spectrums obtained by different basic DOA estim-ators. Even under low SNR conditions, promising DOA estimation performance can be achieved. 相似文献
Layered perovskite Ca2.91Na0.09Ti2-xRhxO7 (x?=?0.00, 0.02, 0.04, 0.06) were synthesized by a conventional solid-state reaction. Room temperature ferroelectricity has been confirmed. The remanent polarization increases with an increase of Rh content, which is due to a larger oxygen octahedral distortion by Rh doping. The coercive field increases with Rh doping as the pinning effect of oxygen vacancies reduce the mobility of domain wall. Remanent polarization and coercive field are caused by different mechanisms, so it is possible to modulate them independently to meet the requirement of application in ferroelectric field. The concentration of oxygen vacancy increased with Rh doping, leading to the significant increase of leakage current density. The bandgap of samples doped with Rh drastically decrease and the visible light response of the sample was improved by Rh doping due to the formation of impurity energy levels within the band gap.