Modulation of mechanosensitive calcium-selective cation channels by temperature

Gating of associations of mechanosensitive Ca²⁺-selective cation co-channels in the plasmalemma of onion epidermis has a strong and unusual temperature dependence. Tension-dependent activity rises steeply as temperature is lowered from 25°C to about 6°C, but drops to a low level at about 5°C. Under the conditions tested (with Mg²⁺ and K⁺ at the cytosolic face of outside-out membrane patches), promotion results both from more bursting at all observed linkage levels and from longer duration of bursts of co-channels linked as quadruplets and quintuplets. Co-channel conductance decreases linearly, but only modestly, with declining temperature. It is proposed that these and related mechanosensitive channels may participate in a variety of responses to temperature, including thermonasty, thermotropism, hydrotropism, and both cold damage and cold acclimation.     

番木瓜果糖-2,6-二磷酸酶CpF2KP基因克隆和蛋白表达纯化

番木瓜是岭南四大名果之一,在我国东南部地区广泛种植,因其具有食用和药用双重价值,因此深受人们的青睐。果糖-6-磷酸,2-激酶/果糖-2,6-二磷酸酯酶(fructose-6-phosphate,2-kinase/fructose-2,6-bisphosphatase,F2KP)是一个独特的双功能酶,具有激酶功能域和酯酶功能域,能催化生物体内糖代谢的重要调节物果糖-2,6-二磷酸(Fru-2,6-P_(2))的合成和降解。为了研究番木瓜中编码该酶的基因CpF2KP的功能,得到目的蛋白尤为重要。本研究从番木瓜基因组中提取到CpF2KP基因的编码序列(coding sequence,CDS)序列,该基因CDS全长2274 bp。将该基因CDS全长扩增之后选用pGEX-4T-1载体进行原核表达。对载体pGEX-4T-1用EcoRⅠ和BamHⅠ进行双酶切,利用基因重组的方式将扩增序列构建到原核表达载体上。经过诱导条件探索,SDS-PAGE结果显示GST-CpF2KP重组蛋白的大小约为110 kDa,诱导CpF2KP蛋白表达的最适条件为:异丙基β-D-硫代半乳糖苷(isopropyl beta-D-thiogalactopyranoside,IPTG)浓度为0.5 mmol/L,温度28℃。对诱导后的CpF2KP蛋白进行纯化,得到了纯化的单一目的蛋白。此外,检测了该基因的组织表达特性发现该基因在种子中表达量最高,在果肉中表达量最低。该研究为进一步深入揭示番木瓜CpF2KP蛋白的功能及研究该基因参与的生物学过程提供了重要基础。     

Impacts of iron on phosphate starvation-induced root hair growth in Arabidopsis

In Arabidopsis, phosphate starvation (-Pi)-induced responses of primary root and lateral root growth are documented to be correlated with ambient iron (Fe) status. However, whether and how Fe participates in -Pi-induced root hair growth (RHG) remains unclear. Here, responses of RHG to different Fe concentrations under Pi sufficiency/deficiency were verified. Generally, distinct dosage effects of Fe on RHG appeared at both Pi levels, due to the generation of reactive oxygen species. Following analyses using auxin mutants and the phr1 mutant revealed that auxin and the central regulator PHR1 are required for Fe-triggered RHG under −Pi. A further proteomic study indicated that processes of vesicle trafficking and auxin synthesis and transport were affected by Fe under −Pi, which were subsequently validated by using a vesicle trafficking inhibitor, brefeldin A, and an auxin reporter, R2D2. Moreover, vesicle trafficking-mediated recycling of PIN2, an auxin efflux transporter, was notably affected by Fe under -Pi. Correspondingly, root hairs of pin2 mutant displayed attenuated responses to Fe under -Pi. Together, we propose that Fe affects auxin signalling probably by modulating vesicle trafficking, chiefly the PIN2 recycling, which might work jointly with PHR1 on modulating -Pi-induced RHG.     

Effects of Allelochemicals on Root Growth and Pod Yield in Response to Continuous Cropping Obstacle of Peanut

Continuous cropping (CC) obstacle is a major threat in legume crops production; however, the underlying mechanisms concerning the roles allelochemicals play in CC obstacle are poorly understood. The current 2-year study was conducted to investigate the effects of different kinds and concentrations of allelochemicals, p-hydroxybenzoic acid (H), cinnamic acid (C), phthalic acid (P), and their mixtures (M) on peanut root growth and productivity in response to CC obstacle. Treatment with H, C, P, and M significantly decreased the plant height, dry weight of the leaves and stems, number of branches, and length of the lateral stem compared with control. Exogenous application of H, C, P, and M inhibited the peanut root growth as indicated by the decreased root morphological characters. The allelochemicals also induced the cell membrane oxidation even though the antioxidant enzymes activities were significantly increased in peanut roots. Meanwhile, treatment with H, C, P, and M reduced the contents of total soluble sugar and total soluble protein. Analysis of ATPase activity, nitrate reductase activity, and root system activity revealed that the inhibition effects of allelochemicals on peanut roots might be due to the decrease in activities of ATPase and NR, and the inhibition of root system. Consequently, allelochemicals significantly decreased the pod yield of peanut compared with control. Our results demonstrate that allelochemicals play a dominant role in CC obstacle-induced peanut growth inhibition and yield reduction through damaging the root antioxidant system, unbalancing the osmolytes accumulation, and decreasing the activities of root-related enzymes.

     

信号分子在生物脱氮中的作用及检测方法

生物脱氮是由微生物主导的地球氮循环中的重要环节之一,主要包括硝化、反硝化和厌氧氨氧化(anaerobic ammonium oxidation,anammox)等过程。在微生物联合作用下,污水中的有机氮及氨氮经一系列作用转化为氮气,这种经济高效、环境友好的处理工艺在世界范围内得到广泛应用。群体感应(quorum sensing,QS)以信号分子为媒介通过改变菌群密度和周围环境变化来调节微生物的各种行为。大量的研究已证实调控QS信号分子在生物脱氮中具有应用潜力。本文介绍了各种信号分子类型,从基因组学、实际应用等方面综述了各类信号分子以及检测方法,同时针对酰基高丝氨酸内酯(acyl homoserine lactones,AHLs)类信号分子在生物脱氮中的作用进行详细介绍。然而不足之处在于信号分子研究只是停留在实验室阶段,仅仅研究了单一信号分子对生物脱氮的影响。未来可将信号分子应用于实际污水,研究多种信号分子共同作用以及多种微生物之间的QS现象。     

定量分析调控人类免疫缺陷病毒潜伏与激活的动力学机制

目的 治疗艾滋病最大的障碍在于无法根除人类免疫缺陷病毒（HIV）潜伏于人体细胞所形成的病毒存储库。构建描述病毒存储库建立分子机制的动力学模型需考虑生物体内的噪声环境和多重影响因素,本文通过一种全新的动力学结构分解方法将随机微分方程的确定性部分与随机性噪声分开,从而在仅需分析常微分方程不动点的情况下即可判断不同药物靶点的作用效果。方法 使用连续的随机微分方程构建了HIV转录过程的动力学模型,简化了描述系统所需方程的维度,增大了模型的可探索空间,在此基础上,通过计算得到的势能函数和概率分布函数直观表示病毒潜伏与激活的不同表达状态以及它们之间的关系。结果 定量分析了不同动力学参数对系统稳态和势函数的影响程度,分别得到了系统处于双稳态和单稳态时的参数范围,并将不同因素对动力系统分岔的影响程度与生物学实验结果对比,验证了本工作的理论基础。结论 本文突破了以往离散、随机的方法,可以通过常微分方程定量分析HIV转录调控的动力学机制,有利于推广到处理高维情况,进一步研究艾滋病在生物体内的发生发展,从而指导设计实验寻找临床上的治疗方案。     

RhoB的翻译后修饰与细胞命运

RhoB作为Rho家族的一员,其生物学活性和蛋白质水平的调控与其他成员有着较大的不同,在肿瘤的发生发展中也起着独特的作用。RhoB作为抑癌蛋白在肿瘤的靶向治疗上受到越来越多的关注,然而在有些类型的肿瘤中RhoB却起着促进肿瘤生长的作用,其中的分子机理还不清楚,亟待研究阐明。可逆的翻译后修饰是快速与精细调控RhoB功能的重要分子机制,对于维持正常细胞的生长、抑制细胞的早期癌变及肿瘤的发生发展至关重要。本文就RhoB翻译后修饰的研究,特别是其泛素化和SUMO化修饰之间的转化在肿瘤细胞命运决定中的作用进行综述,以期为探索RhoB的调控与肿瘤发生发展的机制,以及以RhoB为靶点的癌症治疗提供线索和思路。     

circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移

目的 鼻咽癌是一种来源于鼻咽上皮的恶性肿瘤,其临床特征之一是易发生淋巴转移,但是目前鼻咽癌转移的分子机制尚未阐明。circPVT1是由PVT1基因2号外显子反向拼接形成的环状RNA (circRNA),在多种肿瘤中表达上调,本文探讨了circPVT1在鼻咽癌侵袭迁移中的作用和分子机制。方法 通过RT-qPCR检测circPVT1及其下游miRNA和FSCN1在鼻咽癌细胞的表达情况,Transwell和划痕愈合实验检测circPVT1对鼻咽癌细胞侵袭迁移的影响,RNA pull-down实验检测circPVT1结合的miRNA,双荧光素酶报告实验检测miR-24-3p和let-7a-5p靶向抑制FSCN1 mRNA表达。结果 在鼻咽癌细胞中过表达circPVT1可以促进鼻咽癌细胞侵袭迁移,而敲低circPVT1则可以抑制鼻咽癌细胞的侵袭迁移。进一步研究发现,circPVT1可以通过竞争性吸附miR-24-3p和let-7a-5p,上调FSCN1的表达,从而促进鼻咽癌细胞的侵袭迁移。结论 circPVT1通过miR-24-3p/let-7a-5p/FSCN1轴促进鼻咽癌细胞侵袭迁移,证实c...     

贵州核桃种质资源表型性状多样性研究

该研究以贵州省内的3个山脉(乌蒙山、苗岭山和武陵山)、19个核桃和泡核桃实生居群、245棵实生单株为研究材料,对坚果表型性状29个指标的多样性、相关性、聚类及变异进行分析,以揭示贵州核桃资源的表型遗传多样性水平,为贵州核桃资源的保存与利用及核心种质构建提供依据。结果显示：(1)核桃和泡核桃实生居群245份种质资源的18个表型质量性状Simpson指数为0.26～0.82,Shannon-Wiener指数为0.12～1.79;核桃小叶形状多样,易发生顶叶退化,坚果核壳表面特征、坚果形状及核仁皮色多样性丰富。(2)坚果表型变异系数为3.32%～47.67%,平均为21.28%且差异显著(P<0.05),居群间变异系数为9.42%～31.61%且差异显著(P<0.05);居群内和居群间变异均是核桃表型多样性的来源,但坚果性状在居群间差异显著性均高于居群内,说明居群间变异是该区域坚果表型变异的主要来源。(3)3个山脉区域核桃表型均有不同程度变异,其中,苗岭山核桃表型变异程度低,乌蒙山核桃表型变异程度高;UPGMA聚类结果显示,19个核桃实生居群依据贵州山脉区域划分为三类,表明贵州核...     

内蒙古高寒露天煤矿排土场植被恢复研究

探究不同恢复年限对于露天煤矿植被群落变化和植物个体生长的影响,进一步加强生态环境保护与生态修复治理措施。以内蒙古高寒露天煤矿排土场恢复4年(2018-2021年)的植物群落为研究对象,采用样方调查方法,对样地内植物群落进行调查,设置草本样方(1 m×1 m)和灌木样方(5 m×5 m),记录植物物种组成、高度及盖度等指标,计算Shannon-Wiener多样性指数、Simpon指数、Pielou均匀度指数和物种重要值。探讨不同的恢复年限植物群落变化特征和优势植物的养分吸收,为矿区人工植被恢复提供科学依据。结果表明:(1)矿区排土场植物群落物种数、生物量和群落多样性均随着恢复年限的增加而显著增加。总物种数从恢复第1年的16种增加到恢复第4年的31种,主要是由于非人工种植植物从6种增加到19种导致,特别是在恢复第2年增加显著,然后趋于平稳,这说明恢复第2年是物种增加关键的时期;(2)随着恢复年限的增加,生物量和多样性指数显著增加;矿区植被优势物种多以禾本科披碱草(Elymus dahuricus)、无芒雀麦(Bromus inermis);豆科植物苜蓿(Medicago sativa)、草木樨(Melilotus officinalis)、沙打旺(Astragalus adsurgens)、锦鸡儿(Caragana sinica);菊科植物大籽蒿(Artemisia sieversiana);十字花科油菜(Brassica napus)和胡颓子科沙棘(Hippophae rhamnoides)为主。(3)比较不同优势植物叶片、茎和根的氮(N)、磷(P)和钾(K)含量发现,草本优势种中菊科和豆科植物显著高于禾本科植物。灌木优势种中沙棘茎的N含量和P含量显著高于锦鸡儿。研究认为,在排土场植被恢复初期(4年)的物种选择上,豆科植物和菊科植物是草本植物首选的先锋物种,另外,沙棘是灌木首选的物种。