Production and identification of wheat-Agropyron cristatum 6P translocation lines

The narrow genetic background of wheat is the primary factor that has restricted the improvement of crop yield in recent years. The kernel number per spike is the most important factor of the many potential characteristics that determine wheat yield. Agropyron cristatum (L.) Gaertn., a wild relative of wheat, has the characteristics of superior numbers of florets and kernels per spike, which are controlled by chromosome 6P. In this study, the wheat-A. cristatum disomic addition and substitution lines were used as bridge materials to produce wheat-A. cristatum 6P translocation lines induced by gametocidal chromosomes and irradiation. The results of genomic in situ hybridization showed that the frequency of translocation induced by gametocidal chromosomes was 5.08%, which was higher than the frequency of irradiated hybrids (2.78%) and irradiated pollen (2.12%). The fluorescence in situ hybridization results of the translocation lines showed that A. cristatum chromosome 6P could be translocated to wheat ABD genome, and the recombination frequency was A genome > B genome > D genome. The alien A. cristatum chromosome 6P was translocated to wheat homoeologous groups 1, 2, 3, 5 and 6. We obtained abundant translocation lines that possessed whole-arm, terminal, segmental and intercalary translocations. Three 6PS-specific and four 6PL-specific markers will be useful to rapidly identify and trace the translocated fragments. The different wheat-A. cristatum 6P translocation lines obtained in this study can provide basic materials for analyzing the alien genes carried by chromosome 6P. The translocation line WAT33-1-3 and introgression lines WAI37-2 and WAI41-1, which had significant characteristics of multikernel (high numbers of kernels per spike), could be utilized as novel germplasms for high-yield wheat breeding.     

Preparation,characterization and luminescent properties of dense nano‐silica hybrids loaded with 1,8‐naphthalic anhydride

Novel luminescent dense nano‐silica hybrid materials (DNSS) modified with different amounts of (3‐aminopropyl)triethoxysilane (APTES) and 1,8‐naphthalic anhydride (NA) were successfully synthesized via two steps combined with post‐grafting methods. Powder X‐ray diffraction (XRD), N₂‐sorption analysis, Fourier transform infrared (FT‐IR) spectroscopy, transmission electron microscopy (TEM), thermogravimetric analysis (TGA), photoluminescence (PL) spectroscopy and elemental analysis, as well as time‐resolved decays were employed to characterize the resultant hybrid materials. The results revealed that luminescent organic molecules had been successfully loaded onto the amine‐modified surface of nano‐silica spheres. In addition, their fluorescence intensity and characteristic peak of emission spectra changed with increasing amount of APTES and NA additive. In particular, the characteristic peak showed a red shift from 390 to 450 nm, however, this was inconsistent with results calculated on the basis of the elemental analysis data, most probably because of the dispersion behaviors of NA molecules from the aggregating to the monolayer state. These observations demonstrated the existence of a quantum confinement effectiveness of NA–DNSS samples, and therefore a possible mechanism was put forward. Copyright © 2013 John Wiley & Sons, Ltd.     

Phosphorylation of MtRopGEF2 by LYK3 mediates MtROP activity to regulate rhizobial infection in Medicago truncatula

The formation of nitrogen-fixing no dules on legume roots requires the coordination of infection by rhizobia at the root epidermis with the initiation of cell divisions in the root cortex. During infection, rhizobia attach to the tip of elongating root hairs which then curl to entrap the rhizobia. However, the mechanism of root hair deformation and curling in response to symbiotic signals is still elusive. Here, we found that small GTPases (MtRac1/MtROP9 and its homologs) are required for root hair development and rhizobial infection in Medicago truncatula. Our results show that the Nod factor receptor LYK3 phosphorylates the guanine nucleotide exchange factor MtRopGEF2 at S73 which is critical for the polar growth of root hairs. In turn, phosphorylated MtRopGEF2 can activate MtRac1. Activated MtRac1 was found to localize at the tips of root hairs and to strongly interact with LYK3 and NFP. Taken together, our results support the hypothesis that MtRac1, LYK3, and NFP form a polarly localized receptor complex that regulates root hair deformation during rhizobial infection.     

Arabidopsis chloroplast lipid transport protein TGD2 disrupts membranes and is part of a large complex

In most plants the assembly of the photosynthetic thylakoid membrane requires lipid precursors synthesized at the endoplasmic reticulum (ER). Thus, the transport of lipids from the ER to the chloroplast is essential for biogenesis of the thylakoids. TGD2 is one of four proteins in Arabidopsis required for lipid import into the chloroplast, and was found to bind phosphatidic acid in vitro. However, the significance of phosphatidic acid binding for the function of TGD2 in vivo and TGD2 interaction with membranes remained unclear. Developing three functional assays probing how TGD2 affects lipid bilayers in vitro, we show that it perturbs membranes to the point of fusion, causes liposome leakage and redistributes lipids in the bilayer. By identifying and characterizing five new mutant alleles, we demonstrate that these functions are impaired in specific mutants with lipid phenotypes in vivo. At the structural level, we show that TGD2 is part of a protein complex larger than 500 kDa, the formation of which is disrupted in two mutant alleles, indicative of the biological relevance of this TGD2-containing complex. Based on the data presented, we propose that TGD2, as part of a larger complex, forms a lipid transport conduit between the inner and outer chloroplast envelope membranes, with its N terminus anchored in the inner membrane and its C terminus binding phosphatidic acid in the outer membrane.     

Identification of quantitative trait loci associated with low boron stress that regulate root and shoot growth in Brassica napus seedlings

Brassica napus (Brassicaceae) is among the most important oil crops and a promising biofuel. In the tropics and subtropics, boron (B) deficiency is a major factor limiting Brassica yields. The effect of B on the regulation of root and shoot growth in a doubled haploid (DH) population was evaluated in experiments that utilized hydroponic culture. Strong genetic variability for traits of interest at normal and low B concentrations was demonstrated. Quantitative trait loci (QTL) were analyzed for seven plant growth parameters: increment of primary root length (IPRL), shoot dry weight (SDW), root dry weight (RDW), ratio of RDW to SDW (R/S), shoot B accumulation (SBA), root B accumulation (RBA), and ratio of RBA to SBA [(R/S)BA] in the population. Twenty-seven QTL were detected at normal B levels: four for IPRL, seven for SDW, three for RDW, two for R/S, six for SBA, two for RBA, and three for (R/S)BA. At low B, 18 QTL were detected: four for IPRL, three for SDW, two for RDW, two for R/S, five for SBA, one for RBA, and one for (R/S)BA. Three QTL for adaptability were detected: one A_IPRL and two A_SDW. No putative QTL was detected at both low and normal B. B-related genes were mapped in silico and their locations compared with the QTL identified. The present analyses show the profound and varied effects of B on B. napus and studies on QTL related to B efficiency will help to locate candidate genes and elucidate possible functions of B-efficiency-related QTL.     

解决氧化还原酶反应体系中辅酶问题的策略及其应用

氧化还原酶还原醛或酮生成各种手性醇或手性胺类化合物。然而,氧化还原酶的催化过程通常需要价格昂贵的烟酰胺类辅酶提供或接受电子,这严重阻碍了氧化还原酶的工业化进程。因此,如何降低辅酶的成本已成为生物催化领域的研究热点和关键问题。随着工业化应用的实际需求和研究工作的深入,各种解决辅酶问题的策略被相继提出,如构建体外辅酶再生系统,利用发酵工程与代谢工程等手段提高内源性辅酶利用率,研究和开发辅酶替代物等。文中对这些策略的研究概况进行简要介绍,并通过列举相关应用实例分析各自的优、缺点,为进一步拓展氧化还原酶的工业化应用提供借鉴和参考。     

FGFR1 is essential for N-acetyl-seryl-aspartyl-lysyl-proline regulation of mitochondrial dynamics by upregulating microRNA let-7b-5p

Fibroblast growth factor receptor (FGFR) 1 plays a key role in endothelial homeostasis by inducing microRNA (miR) let-7. Our previous paper showed that anti-fibrotic effects of N-acetyl-seryl-aspartyl-lysyl-proline (AcSDKP) were associated with restoring diabetes-suppressed expression of FGFR1 and miR let-7, the key contributor of mitochondrial biogenesis, which is regulated by mitochondrial membrane GTPase proteins (MFN2 and OPA1). Here, we found that the FGFR1 signaling pathway was critical for AcSDKP in maintaining endothelial mitochondrial biogenesis through induction of miR let-7b-5p. In endothelial cells, AcSDKP restored the triple cytokines (TGF-β2, interleukin-1β, tumor necrosis factor-α)-suppressed miR let-7b-5p and protein levels of the mitochondrial membrane GTPase. This effect of AcSDKP was lost with either fibroblast growth factor receptor substrate 2 (FRS2) siRNA or neutralizing FGFR1-treated cells. Similarly, AcSDKP had no effect on the miR let-7b-5p inhibitor-suppressed GTPase levels in endothelial cells. In addition, a miR let-7b-5p mimic restored the levels of FRS2 siRNA-reduced GTPases in endothelial cells. These findings were also confirmed using MitoTracker Green and an immunofluorescence assay. Our results demonstrated that the AcSDKP-FGFR1 signaling pathway is critical for maintaining mitochondrial dynamics by control of miR let-7b-5p in endothelial cells.     

Substrate,Hormone, Winnowing,and Stratification Influence the Seed Germination of <i>Ilex asprella</i> (Hook. et Arn.) Champ. ex Benth

Ilex asprella (Hook. et Arn.) Champ. ex Benth is one of the most important traditional Chinese medicines in southern China. The seeds of Ilex asprella usually have extremely low germination due to their dormancy characteristics, which severely impacts the efficiency of seedling raising and increases labor costs. In this study, to improve the seed germination of I. asprella, the effects of germination substrate, hormone, winnowing, and stratification treatments on the seed germination of I. asprella were investigated. The results of the germination matrix showed that the highest germination percentage of 45.2% was achieved under the 20°C/10°C day/night temperature and vermiculite germination medium conditions. The results of hormone treatments revealed that 100–400 mg/L of gibberellin (GA) and 50–100 mg/L of salicylic acid (SA) were found to be effective in releasing the dormancy of I. asprella seeds. Moreover, winnowing could effectively eliminate unsaturated seeds and impurities, thus improving the seed germination of I. asprella. Furthermore, warm temperature (15°C) stratification could expand the temperature range of I. asprella’s seed germination, which was beneficial for seed germination of I. asprella and for seed nursery at room temperature in production practice. The present study obtained a method to break dormancy and increase seed germination in I. asprella, thereby forming a groundwork for improving the efficiency of large-scale planting of I. asprella.     

氯胺酮对川金丝猴的麻醉效应

本研究单独使用氯胺酮对24只神农架川金丝猴（Rhinopithecus roxellana）进行麻醉,比较其在诱导期、麻醉期及恢复期的持续时间和行为反应差异,探讨氯胺酮对不同性别、不同年龄川金丝猴的麻醉效果。结果显示：24例受试动物注射氯胺酮（剂量5.90-15.71mg/kg）后,镇痛、镇静和肌松效果良好,无意外死亡。其中,诱导期表现四肢逐渐无力,流涎,身体逐渐倒下,偶有嘴空嚼;麻醉期表现肌松良好,呼吸通畅,伴有嘴空嚼;恢复期表现以手指和躯干振动,开始恢复意识,嘴空嚼情况增多,尝试坐立和行走,最后恢复正常;诱导期、麻醉期和恢复期持续平均持续时间分别为(4.00 ± 1.77)min、(21.67 ± 5.03)min和(31.79 ± 6.07)min。本研究首次监测到川金丝猴在麻醉过程中出现嘴空嚼行为（没有进食的情况下嘴上下及左右咀嚼）。数据分析表明,在麻醉过程中雄性个体恢复期时长明显大于雌性,雄性个体在恢复期空嚼行为的次数明显小于雌性;而性别对诱导期、麻醉期,年龄对诱导期、麻醉期和恢复期的时长及行为影响均无明显差异。