棒状腐败乳杆菌Lc7的生物学特性及益生作用

【目的】对分离自健康成人粪便样本的棒状腐败乳杆菌(Loigolactobacillus coryniformis)Lc7进行分类学鉴定和益生潜力评估。【方法】基于16SrRNA基因和基因组核心基因构建系统发育树,对Lc7进行分类学鉴定;通过耐酸和胆汁酸盐、粘附、抗氧化和抑菌实验,以及溶血、明胶酶活性和抗菌药物敏感性实验,评估Lc7的益生特性。同时,构建小鼠溃疡性结肠炎模型,评估Lc7的体内抗炎潜力。【结果】Lc7鉴定为L. coryniformis,在酸和胆汁酸盐的连续作用下,Lc7的存活率为70.17%。Lc7对HT-29细胞的粘附指数为56.33 CFU/cell,其自聚集和疏水性分别为80%和40%;Lc7对福氏志贺菌和鼠伤寒沙门菌等7个常见致病菌均有较强的抑制能力;对1,1-二苯基-2-苦基肼(1,1-diphenyl-2-picryl-hydrazyl,DPPH)和羟自由基(hydroxyl radicals,·OH)的清除率分别为91.70%和48.53%;Lc7无溶血现象和明胶酶活性,对选取的大多数抗生素均敏感。在小鼠结肠炎实验中,Lc7干预组小鼠结肠长度明显长于模型组(...     

The potato StMKK5-StSIPK module enhances resistance to Phytophthora pathogens through activating the salicylic acid and ethylene signalling pathways

Mitogen-activated protein kinase (MAPK) cascades play pivotal roles in plant responses to both biotic and abiotic stress. A screen of a Nicotiana benthamiana cDNA virus-induced gene silencing (VIGS) library for altered plant responses to inoculation with Phytophthora infestans previously identified an NbMKK gene, encoding a clade D MAPKK that we renamed as NbMKK5, which is involved in immunity to P. infestans. To study the role of the potato orthologous gene, referred to as StMKK5, in the response to P. infestans, we transiently overexpressed StMKK5 in N. benthamiana and observed that cell death occurred at 2 days postinfiltration. Silencing of the highly conserved eukaryotic protein SGT1 delayed the StMKK5-induced cell death, whereas silencing of the MAPK-encoding gene NbSIPK completely abolished the cell death response. Further investigations showed that StMKK5 interacts with, and directly phosphorylates, StSIPK. Furthermore, both StMKK5 and StSIPK trigger salicylic acid (SA)- and ethylene (Eth)-related gene expression, and co-expression of the salicylate hydroxylase NahG with the negative regulator of Eth signalling CTR1 hampers StSIPK-triggered cell death. This observation indicates that the cell death triggered by StMKK5-StSIPK is dependent on the combination of SA- and Eth-signalling. By introducing point mutations, we showed that the kinase activity of both StMKK5 and StSIPK is required for triggering cell death. Genetic analysis showed that StMKK5 depends on StSIPK to trigger plant resistance. Thus, our results define a potato StMKK5-SIPK module that positively regulates immunity to P. infestans via activation of both the SA and Eth signalling pathways.     

TaTPP-7A positively feedback regulates grain filling and wheat grain yield through T6P-SnRK1 signalling pathway and sugar–ABA interaction

Grain size and filling are two key determinants of grain thousand-kernel weight (TKW) and crop yield, therefore they have undergone strong selection since cereal was domesticated. Genetic dissection of the two traits will improve yield potential in crops. A quantitative trait locus significantly associated with wheat grain TKW was detected on chromosome 7AS flanked by a simple sequence repeat marker of Wmc17 in Chinese wheat 262 mini-core collection by genome-wide association study. Combined with the bulked segregant RNA-sequencing (BSR-seq) analysis of an F₂ genetic segregation population with extremely different TKW traits, a candidate trehalose-6-phosphate phosphatase gene located at 135.0 Mb (CS V1.0), designated as TaTPP-7A, was identified. This gene was specifically expressed in developing grains and strongly influenced grain filling and size. Overexpression (OE) of TaTPP-7A in wheat enhanced grain TKW and wheat yield greatly. Detailed analysis revealed that OE of TaTPP-7A significantly increased the expression levels of starch synthesis- and senescence-related genes involved in abscisic acid (ABA) and ethylene pathways. Moreover, most of the sucrose metabolism and starch regulation-related genes were potentially regulated by SnRK1. In addition, TaTPP-7A is a crucial domestication- and breeding-targeted gene and it feedback regulates sucrose lysis, flux, and utilization in the grain endosperm mainly through the T6P-SnRK1 pathway and sugar–ABA interaction. Thus, we confirmed the T6P signalling pathway as the central regulatory system for sucrose allocation and source–sink interactions in wheat grains and propose that the trehalose pathway components have great potential to increase yields in cereal crops.     

不同气候区绿色屋顶蒸散发模拟研究

蒸散发过程决定绿色屋顶雨水滞留能力的恢复,进而影响绿色屋顶径流调控功能。基于水量平衡原理和Penman-Monteith公式,利用北京市实验绿色屋顶气象和蒸散发连续监测数据,构建并验证绿色屋顶水文过程模型,模拟分析不同气候区城市绿色屋顶蒸散发变化规律。结果表明：(1)该模型能较准确模拟绿色屋顶蒸散发量,率定和检验期的Nash-Sutcliffe效率系数分别为0.6385和0.6014,决定系数(R²)分别为0.7191和0.6168;(2)基质厚度相同的情况下,从半干旱区(兰州)、半湿润区(北京)到湿润区(武汉和广州),绿色屋顶日平均实际蒸散发量呈增加趋势;(3)增加基质厚度可提升绿色屋顶最大雨水滞留能力,进而增加绿色屋顶实际蒸散发量,但基质厚度对绿色屋顶蒸散发量的影响存在阈值,在兰州、北京、武汉和广州,当基质厚度分别超过10 cm、17 cm、24 cm和25 cm时,绿色屋顶的日平均实际蒸散发量变化不再明显。此外,不同气候区城市绿色屋顶的日平均实际蒸散发量也存在阈值,广州绿色屋顶日平均实际蒸散发量的阈值依次高于武汉、北京和兰州。本研究有望为我国不同气候区绿色屋...     

关帝山云杉次生林树木径向生长的空间关联及其影响因子

以关帝山4 hm²云杉次生林样地为研究对象,按照CTFS(Center for Tropical Forest Science)技术规范对样地树木进行连续定位监测。利用2010至2015年间样地主要树种生长量观测数据,结合地形、土壤等环境因子调查及采样测定数据,分析了树木种群径向生长的空间关联性及其随生境的变化,并探讨了树木种群径向生长的影响因素。结果表明,青杄、华北落叶松、红桦、白桦和辽东栎为云杉次生林主要树种,在样地4个生境型(山脊生境、低海拔缓坡生境、高海拔缓坡生境、低洼地陡坡生境)中均有分布且呈现不同的径级结构。标记相关函数分析显示,同一生境型中,5树种径向生长的空间关联性各异;对于同一树种,径向生长的空间自相关性不仅具有尺度依赖性,同时生境型的不同导致树木径向生长的空间关联性发生变化。线性混合效应模型分析显示,初始胸径对树木径向生长的显著正效应在样地各类生境型的所有种群中普遍存在;生物因子对树木径向生长的显著影响只在特定生境型的青杄种群中被检测到,表明树木径向生长受同种邻体影响,但其影响显著性因树种而异;环境因子中,海拔和凹凸度对树木径向生长呈显著负效应...     

酶法生产L-高苯丙氨酸的研究进展

L-高苯丙氨酸(L-homophenylalanine,L-HPA)作为一种重要的非天然氨基酸,是合成治疗高血压的普利类药物等的关键中间体,具有广阔的市场前景。目前L-高苯丙氨酸的合成主要依赖于化学法,但化学合成L-高苯丙氨酸具有原料昂贵、步骤繁琐和污染严重等缺点,限制了广泛应用。因此,国内外研究者对L-高苯丙氨酸的酶法生产进行了深入的研究。本文就目前酶法合成L-高苯丙氨酸的工艺,包括脱氢酶法、转氨酶法、海因酶法和脱羧酶法的研究进展进行了综述,为酶法合成L-高苯丙氨酸提供一定的借鉴,为最终实现L-高苯丙氨酸的酶法工业化生产奠定基础。     

谷氨酸棒杆菌代谢工程高效生产L-缬氨酸北大核心CSCD

L-缬氨酸作为一种支链氨基酸,广泛应用于医药和饲料等领域。本研究借助多种代谢工程策略相结合的方法,构建了生产L-缬氨酸的微生物细胞工厂,实现了L-缬氨酸的高效生产。首先,通过增强糖酵解途径、减弱副产物代谢途径相结合的方式,强化了L-缬氨酸合成前体丙酮酸的供给;其次,针对L-缬氨酸合成路径关键酶—乙酰羟酸合酶进行定点突变,提高了菌株的抗反馈抑制能力,并利用启动子工程策略,优化了路径关键酶的基因表达水平;最后,利用辅因子工程策略,改变了乙酰羟酸还原异构酶和支链氨基酸转氨酶的辅因子偏好性,由偏好NADPH转变为偏好NADH,从而提高了L-缬氨酸的合成能力。在5L发酵罐中,最优谷氨酸棒杆菌工程菌株Corynebacterium glutamicum K020的L-缬氨酸产量、得率和生产强度分别达到了110g/L、0.51g/g和2.29 g/(L·h)。     

Genetic variation in the Y chromosome and sex-biased DNA methylation in somatic cells in the mouse

Mammalian Genome - Several lines of evidence suggest that the presence of the Y chromosome influences DNA methylation of autosomal loci. To better understand the impact of the Y chromosome on...     

Nonparametric and semiparametric estimation with sequentially truncated survival data

In observational cohort studies with complex sampling schemes, truncation arises when the time to event of interest is observed only when it falls below or exceeds another random time, that is, the truncation time. In more complex settings, observation may require a particular ordering of event times; we refer to this as sequential truncation. Estimators of the event time distribution have been developed for simple left-truncated or right-truncated data. However, these estimators may be inconsistent under sequential truncation. We propose nonparametric and semiparametric maximum likelihood estimators for the distribution of the event time of interest in the presence of sequential truncation, under two truncation models. We show the equivalence of an inverse probability weighted estimator and a product limit estimator under one of these models. We study the large sample properties of the proposed estimators and derive their asymptotic variance estimators. We evaluate the proposed methods through simulation studies and apply the methods to an Alzheimer's disease study. We have developed an R package, seqTrun , for implementation of our method.