Real Time Measures of Prestin Charge and Fluorescence during Plasma Membrane Trafficking Reveal Sub-Tetrameric Activity

Prestin (SLC26a5) is the outer hair cell integral membrane motor protein that drives cochlear amplification, and has been described as an obligate tetramer. We studied in real time the delivery of YFP-prestin to the plasma membrane of cells from a tetracycline-inducible cell line. Following the release of temperature block to reinstate trans Golgi network delivery of the integral membrane protein, we measured nonlinear capacitance (NLC) and membrane fluorescence during voltage clamp. Prestin was delivered exponentially to the plasma membrane with a time constant of less than 10 minutes, with both electrical and fluorescence methods showing high temporal correlation. However, based on disparity between estimates of prestin density derived from either fluorescence or NLC, we conclude that sub-tetrameric forms of prestin contribute to our electrical and fluorescence measures. Thus, in agreement with previous observations we find that functional prestin is not an obligate tetramer.     

主养青鱼池塘生态系统能量转换率的研究

1985—1987年对苏州市郊区主养青鱼池塘生态系统的能量转换率进行了分析。结果表明,主养青鱼净产7.5、11.25、15t/ha 3个产量级型池塘青鲤团头鲂产出能占养鱼总产出能的比例分别为82.49、78.03、79.34％;总投入能(太阳辐射能+辅助能)转移到鱼的总产出能转换率分别为0.19、0.24、0.31％;太阳辐射能转移到毛和净初级生产力的能量转换率分别为0.76、0.90、0.96％和0.61、0.72、0.77％;净初级生产力转移到滤食性鱼净产量的能量转换率分别为4.02、4.63、5.27％;辅助能转移到鱼净产量的能量转换率分别为12.20、11.33。11.74％。在3个产量级型池塘中,以15t/ha产量级的能量转换率为最佳型。     

皖南、赣北奥陶纪笔石立体标本形成环境的初步研究*

皖南、赣东北和赣西北地区奥陶纪笔石地层发育良好,笔石化石丰富。宁国组和胡乐组均为笔石相地层,但笔石的保存特点并不相同。立体保存的黄铁矿化笔石标本主要见于宁国组,而胡乐组的笔石几乎均为薄膜标本。在比较宁国组和胡乐组在岩性、颜色、化石、矿物和元素等方面的特点后发现,两者有较明显的差异。这表明宁国组和胡乐组形成时的环境是不同的,前者为弱还原环境,后者为较强的还原环境,而在研究区内影响笔石体立体保存的主要因素为还原环境和较高的铁含量。在还原环境下,铁可呈Fe~(2+)存在,笔石体内含有硫,死亡后经降解作用可生成H_2S;H_2S和Fe~(2+)结合可使笔石体黄铁矿化,从而使笔石体硬化而呈立体保存下来。宁国组的铁含量明显高于胡乐组,这似可以解释宁国组产有较多笔石立体标本的原因。     

声学指数在城市森林鸟类多样性评估中的应用

鸣声是鸟类之间进行沟通和传递信息的重要方式,这为通过声学监测评估鸟类多样性提供了独特的机会。利用声学指数快速评估生物多样性是一种新兴的调查方法,但城市森林中的复杂声环境可能会导致声学指数的指示结果出现偏差。为了解声学指数在城市森林中应用的可行性,本研究在北京市东郊森林公园设置了50个矩阵式调查样点,于2021年4–6月每月进行1次鸟类传统观测和同步鸣声采集,通过比较两种方法的结果来探究声学监测的有效性。采用Spearman相关分析和广义线性混合模型评估6个常用声学指数与鸟类丰富度和多度的关系,并衡量了每个指数的性能。结果表明:(1)本研究共记录到鸟类10目23科35种,通过声学监听识别的总物种数与传统鸟类观测相等,但具体鸟种存在差异;(2)不同月份间声学指数与鸟类丰富度和多度的相关性有明显差别,声学复杂度指数(ACI)和标准化声景差异指数(NDSI)优于其他指数,是评估鸟类多样性的关键变量;(3)声学指数对鸟类多度的预测能力(R²m=0.32,R2c=0.80)要高于丰富度(R²m=0.12,R2c=0.18)。声学指数为快速评估生物多样性提...     

High-resolution mapping of the S and Z loci of Phalaris coerulescens.

Self incompatibility (SI) in Phalaris coerulescens is gametophytically determined by two unlinked multi allelic loci (S and Z). Neither the S nor Z genes have yet been cloned. As part of a map-based cloning strategy, high-resolution maps of the S and Z regions were generated from distorted segregating populations using RFLP probes from wheat, barley, oat, and Phalaris. The S locus was delimited to 0.26 cM with two boundary markers (Xwg811 and Xpsr168) and cosegregated with Xbm2 and Xbcd762. Xbcd266 was the closest marker linked to Z (0.9 cM). A high level of colinearity in the S and Z regions was found in both self-incompatible and -compatible species. The S locus was localized to the subcentromere region of chromosome 1 and the Z locus to the long arm end of chromosome 2. Several rice BAC clones orthologous to the S and Z locus regions were identified. This opens the possibility of using the rice genome sequence data to generate more closely linked markers and identify SI candidate genes. These results add further support to the conservation of gene order in the S and Z regions of the grass genomes.     

FK-506结合蛋白对钙释放通道的调控

细胞内自由钙作为一种重要的细胞信使广泛地参与细胞生理功能调控.胞内钙库(内质网系和肌浆网系)对调节细胞内自由钙水平起着重要的作用.钙库膜上的钙释放通道(ryanodine受体和三磷酸肌醇受体)受许多因素调控,其中之一就是新近研究得相当多的FK506结合蛋白.免疫抑制剂FK506能特异地结合钙库上一种分子质量为12 ku左右的蛋白,这种FK506结合蛋白与钙释放通道形成一种紧密连接的复合体,在正常生理情况下对钙释放通道起着十分重要的调控作用.     

Developmental control of xylem hydraulic resistances and vulnerability to embolism in Fraxinus excelsior L.: impacts on water relations

The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 10^–4M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar ‘Alaska’, an ABA-deficientmutant ‘wil’, and its ‘wildtype’. Freezinginjury was determined graphically as the temperature that caused50% injury (T₅₀) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T₅₀ for root tissuewas 1C for all genotypes, regardless of treatment. For ‘Alaska’shoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by –1.5 to –4.0C, while coldtreatment increased the freezing tolerance by –7.5 to–14.8C. Cold treatment increased the freezing toleranceof shoot tips by –9 and –15C for ‘wil’and ‘wild-type’, respectively. Cold acclimationincreased endogenous ABA concentrations in ‘Alaska’shoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of ‘Alaska’, respectively.In ‘Alaska’ shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in ‘wil’and ‘wild-type’ shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization     

Phase separation in short-chain lecithin/gel-state long-chain lecithin aggregates

Small bilayer particles form spontaneously from gel-state long-chain phospholipids such as dipalmitoylphosphatidylcholine and 0.2 mol fraction short-chain lecithins (e.g., diheptanoyl-phosphatidylcholine). When the particles are incubated at temperatures greater than the Tm of the long-chain phosphatidylcholine (PC), the particles rapidly fuse (from 90-A to greater than or equal to 5000-A radius); this transition is reversible. A possible explanation for this behavior involves patching or phase separation of the short-chain component within the gel-state particle and randomization of both lipid species above Tm. Differential scanning calorimetry, 1H T1 values of proteodiheptanoyl-PC in diheptanoyl-PC-d26/dipalmitoyl-PC-d62 matrices of varying deuterium content, solid-state 2H NMR spectroscopy as a function of temperature, and fluorescence pyrene excimer-to-monomer ratios as a function of mole fraction diheptanoyl-PC provide evidence that such phase separation must occur. These results are used to construct a phase diagram for the diheptanoyl-PC/dipalmitoyl-PC system, to propose detailed geometric models for the different lipid particles involved, and to understand phospholipase kinetics toward the different aggregates.     

Application and future perspective of CRISPR/Cas9 genome editing in fruit crops

Fruit crops, including apple, orange, grape,banana, strawberry, watermelon, kiwifruit and tomato, not only provide essential nutrients for human life but also contribute to the major agricultural output and economic growth of many countries and regions in the world. Recent advancements in genome editing provides an unprecedented opportunity for the genetic improvement of these agronomically important fruit crops. Here, we summarize recent reports of applying CRISPR/Cas9 to fruit crops,including efforts to reduce disease susceptibility, change plant architecture or flower morphology, improve fruit quality traits, and increase fruit yield. We discuss challenges facing fruit crops as well as new improvements and platforms that could be used to facilitate genome editing in fruit crops, including d Cas9-base-editing to introduce desirable alleles and heat treatment to increase editing efficiency. In addition, we highlight what we see as potentially revolutionary development ranging from transgene-free genome editing to de novo domestication of wild relatives. Without doubt, we now see only the beginning of what will eventually be possible with the use of the CRISPR/Cas9 toolkit. Efforts to communicate with the public and an emphasis on the manipulation of consumerfriendly traits will be critical to facilitate public acceptance of genetically engineered fruits with this new technology.     

Intersected functional zone of transcriptional regulators patterns stemness within stem cell niche of root apical meristem

Root stem cell niche (SCN) consists of a quiescent center (QC) and surrounding stem cells. Disrupted symplastic communication leads to loss of stemness in the whole SCN. Several SCN regulators were reported to move between cells for SCN maintenance. However, single mutant of these regulators is insufficient to abolish QC stemness despite the high differentiation rate in surrounding stem cells. To dissect the mechanism behind such distinct stemness in SCN, we combined the mis‐expression strategy with pWOX5:icals3m system in which QC is symplastically isolated. We found the starch accumulation in QC could be synergistically repressed by WUSCHEL‐RELATED HOMEOBOX 5 (WOX5), SHORT‐ROOT (SHR), SCARCROW (SCR), and PLETHORA (PLT). Like PLTs, other core regulators also exhibited dimorphic functions by inhibiting differentiation at a higher dose while promoting cell division at a low protein level. Being located in the center of the intersected expression zones, QC cells receive the highest level of core regulators, forming the most robust stemness within SCN. WUSCHEL‐RELATED HOMEOBOX 5 was sufficient to activate PLT1/2 expression, contributing to the QC‐enriched PLTs. Our results provide experimental evidence supporting the long‐standing hypothesis that the combination of spatial expression, synergistic function and dosage effect of core regulators result in spatially distinct stemness in SCN.