Seed and microsite limitation in Rheum nobile,a rare and endemic plant from the subnival zone of Sino-Himalaya

Background: Species persistence, particularly in monocarpic species, depends on the successful recruitment of individuals. An understanding of the factors that limit the recruitment of rare monocarpic plant species is therefore vital for their conservation.

Aims: To identify the factors limiting the recruitment of Rheum nobile, a rare and highly specialised monocarpic giant herb endemic to the high eastern Himalayas.

Methods: Seed sowing (seeds added or not added) and seedling transplanting experiments were conducted in disturbed (vegetation removed) and undisturbed plots in the vicinity of established populations of R. nobile to explore the mechanisms of recruitment limitation. Four levels of photosynthetically active radiation (0, 15, 30 and 50 μmol m^?2 s^?1) and two sowing positions (beneath and above grass litter or moss layer) were manipulated in the laboratory to determine how ground cover limited seedling emergence.

Results: Seed addition increased seedling recruitment. Disturbance significantly increased seedling emergence and establishment. Seed germination significantly decreased with the reduction of light availability, but 31.7% of all seeds germinated in complete darkness. Seedling emergence was close to zero when seeds were sown on top of a layer of grass litter or moss, but rose to 34.5% when the seeds were sown beneath such layers.

Conclusions: Our results indicate that the recruitment of R. nobile is limited by a combination of seed and microsite availability. Therefore, in order to conserve this species, we suggest adding seeds to suitable sites and implementing soil disturbances in existing populations to create suitable microsites.     

Process intensification in fed-batch production bioreactors using non-perfusion seed cultures

ABSTRACT

Although process intensification by continuous operation has been successfully applied in the chemical industry, the biopharmaceutical industry primarily uses fed-batch, rather than continuous or perfusion methods, to produce stable monoclonal antibodies (mAbs) from Chinese hamster ovary (CHO) cells. Conventional fed-batch bioreactors may start with an inoculation viable cell density (VCD) of ~0.5 × 10⁶ cells/mL. Increasing the inoculation VCD in the fed-batch production bioreactor (referred to as N stage bioreactor) to 2–10 × 10⁶ cells/mL by introducing perfusion operation or process intensification at the seed step (N-1 step) prior to the production bioreactor has recently been used because it increases manufacturing output by shortening cell culture production duration. In this study, we report that increasing the inoculation VCD significantly improved the final titer in fed-batch production within the same 14-day duration for 3 mAbs produced by 3 CHO GS cell lines. We also report that other non-perfusion methods at the N-1 step using either fed batch or batch mode with enriched culture medium can similarly achieve high N-1 final VCD of 22–34 × 10⁶ cells/mL. These non-perfusion N-1 seeds supported inoculation of subsequent production fed-batch production bioreactors at increased inoculation VCD of 3–6 × 10⁶ cells/mL, where these achieved titer and product quality attributes comparable to those inoculated using the perfusion N-1 seeds demonstrated in both 5-L bioreactors, as well as scaled up to 500-L and 1000-L N-stage bioreactors. To operate the N-1 step using batch mode, enrichment of the basal medium was critical at both the N-1 and subsequent intensified fed-batch production steps. The non-perfusion N-1 methodologies reported here are much simpler alternatives in operation for process development, process characterization, and large-scale commercial manufacturing compared to perfusion N-1 seeds that require perfusion equipment, as well as preparation and storage vessels to accommodate large volumes of perfusion media. Although only 3 stable mAbs produced by CHO cell cultures are used in this study, the basic principles of the non-perfusion N-1 seed strategies for shortening seed train and production culture duration or improving titer should be applicable to other protein production by different mammalian cells and other hosts at any scale biologics facilities.     

Studies on the Utilization of Hydrocarbons by Microorganisms

In the course of investigation of alicyclic hydrocarbon-utilizing microorganisms, five strains of ethylcyclohexane-utilizing bacteria were isolated from soil samples.

Among those bacteria, the strain S6B1 that was identified as Alcaligenes faecalis, showed the best growth in shaking culture.

The strain S6B1 was found to produce 4-ethylcyclohexanol from ethylcyclohexane.

This substance separated from culture broth was purified and identified to be trans-4-ethylcyclohexanol by the use of NMR.     

火烧信号对种子萌发影响的研究进展

火作为一个基础的生态因子, 对森林、草地等陆地生态系统的结构和功能有着重要的影响。种子萌发是种子植物的重要生活史阶段, 也是火后植被更新和恢复的主要途径。植被燃烧产生了烟、热以及与烟相关的一系列火烧信号, 在打破种子休眠, 促进种子萌发方面发挥重要作用。该文将火烧信号分为物理信号和化学信号, 物理信号主要是伴随火烧产生的高温, 化学信号主要包括气态烟以及近年来从烟水中提取的影响种子萌发的关键化学物质karrikins和glyceronitrile。该文围绕火烧的基本信息, 火烧信号对种子萌发的影响, 火烧信号在实践中的应用3个方面进行系统综述, 重点探讨了不同类别的火烧信号及其交互作用对种子萌发的影响。在系统总结火烧信号对种子萌发影响的研究进展的基础上, 提出未来的研究应与烟信号作用机理的探究以及全球变化等方面相结合, 旨在充分发挥火的生态服务功能, 为火的科学管理应用和退化生态系统恢复提供理论支撑。     

麻楝种子形态和营养特征遗传变异分析

为系统揭示麻楝种源间种子形态和营养成分的变异程度和变异规律,以8个国家26个种源麻楝种子为试验材料,对不同种源麻楝种子的形态特征和营养成分进行比较分析。多重比较结果表明,麻楝种源间种子的长、宽、厚、长宽比、百粒重以及可溶性蛋白质、可溶性糖和淀粉含量差异极显著;相关分析表明,种子百粒重与种子长、种子宽呈极显著正相关,可溶性蛋白质、可溶性糖和淀粉含量与百粒重均呈极显著正相关;主成分分析结果显示,影响麻楝种子品质的第1主成分主要是种子形态性状,即种子长、百粒重和种子宽,第2主成分是种子营养成分;通过分析麻楝种子形态特征及营养成分变异系数可知,麻楝种子形态变异系数大于营养成分变异系数,表明种子形态受外界环境影响较大。通过主成分聚类分析,将26个种源分为2个集群,第1集群包括澳大利亚和马来西亚2个种源,其特点是种子形态较大、百粒较重、养分含量较高;第2集群包括其余24个种源。结合聚类分析和种子形态与地理气候因子相关分析结果可知,麻楝种子形态变异具有一定的地理区域特征,与分布区的自然地理因素有一定的相关性,麻楝分布区南部种子较大而北部种子较小。     

江西三清山濒危植物天女花种子特性研究

研究了江西三清山天女花种子的特性,结果表明：(1)三清山天女花种群种子雨自10月初开始,10月末结束,10月中旬强度达高峰,玉京峰地段天女花种子雨持续时间较短;(2)产种率受母树冠层影响,即上层>中层>下层;种子败育率高(49.4%～100.0%);(3)主成分分析表明,乔木层盖度、大气湿度、光照因子以及土壤含水量对天女花种子发育产生显著影响。     

The effect of frugivory on postdispersal seed removal and germination in the pantropical forest tree Antiaris toxicaria Leschenault

The quality of seed treatment by frugivores has an effect on seed removal after dispersal, seed germination and tree recruitment. We provide information on postdispersal seed removal, germination and subsequent recruitment in tropical forest tree species Antiaris toxicaria in Ghana. We tested whether postdispersal seed removal and germination rates were differentially affected by the following seed treatments: seeds that were spat out by monkeys with all fruit pulp removed and spitting seeds with fruit pulp partially removed as observed in some birds and bats. We used seeds of intact ripened fruits as control. Frugivore seed treatment and distance from bole affected seed removal patterns, whereas intact seeds were significantly removed from all seed stations. The germination success was greater for seeds that were spat out by monkeys and poor for seeds with fruit pulp partially removed and intact fruits. More recruits were recorded at the edge of the adult A. toxicaria canopy radius. There was weak relationship (r² = 0.042) between the number of recruits and distance away from the adult tree. Results suggest that the subsequent recruitment in tropical forest tree species may be enhanced by some frugivore fruit‐handling behaviour where fruit pulp is removed from the seeds without destroying the seeds.     

侧柏不同器官水提取物对油松种子萌发和幼苗生长的他感效应

研究了侧柏（Platycladus orientalis）根、枝、叶、果实不同浓度水提取物及枝叶挥发油对油松种子萌发和幼苗生长的他感效应。结果表明：侧柏各器官及挥发油都含有化感物质,均对油松种子萌发及幼苗生长表现出“高抑低促”效应,而且低浓度时的促进作用明显强于高浓度时的抑制作用。与对照相比,在浓度为2 mg·mL^-1时,侧柏根水提取物对油松幼苗根长和鲜重的促进能力最强,其根长和鲜重分别比对照提高了79.78%和376.60%;侧柏叶水提取物对油松幼苗苗高促进能力最强,比对照提高了102.41%;侧柏枝水提取物对油松种子发芽的促进能力最强,发芽率比对照提高了65.63%。而在浓度为30 mg·mL^-1时,除了侧柏根和枝水提取物对油松种子萌发（87.50%）和鲜重（85.11%）表现出明显的抑制作用外,其它提取物对油松幼苗根长和苗高生长的抑制作用均不明显。侧柏枝叶挥发油对油松种子萌发,幼苗生长均表现为促进作用,特别对油松种子萌发具有显著的促进作用,其促进率达70.37%。说明侧柏和油松混交,可能有利于油松树木生长及生产力的提高。     

影响4类作物种子寿命的主要贮藏环境因素

为研究利用存活比、影响种子贮藏寿命和生活力的主要贮藏环境因子来预测高粱、苋属、甜荞麦和莴苣种子批的寿命和生活力,3个含水量水平的高粱和苋属的8个种子批被贮藏在20℃、30℃和40℃的贮藏温度下长达810 d。以概率回归和存活比线性回归方法,贮藏试验获得的高粱、苋属及文献报道的甜荞麦、莴苣种子批的发芽率和贮藏时间的成套数据,被用于估算不同种子批的平均贮藏寿命(p50)和种子死亡时间分布的标准差(σ),同时两种回归方程的拟合性也被比较;两种方法估算出的p50和σ则分别被用于对贮藏环境因子:贮藏温度(t)及其平方、种子含水量(m)及其对数(logm)以及t和m的交互作用(t×m和t×logm)作逐步回归分析,结果表明,用存活比来估算p50、σ是可行的,而且估算出的p50和σ较用概率估算的有统计学意义;贮藏温度和种子含水量的交互作用(t×m)是使4类作物种子快速丧失生活力,减少寿命的主要贮藏环境因子,包含这个因子和存活比的数学模型:G i/G0=A-p/10K-CWT(t×m),预测甜荞麦种子批的发芽率较V i=K i-p/10K-CWT(t×m)有更高的准确性;所以,建议用前者来预测初始发芽率是已知的种子的贮藏寿命和生活力,而用后者预测初始发芽率未被测定的种子的寿命和生活力。