Conservation,propagation, and redistribution (CPR) of Hill’s thistle: paradigm for plant species at risk

Cryopreservation is a valuable tool that could potentially create an alternate plant preservation strategy for species at risk such as Hill’s thistle. The present study is focused on a successful paradigm involving conservation, propagation and redistribution (CPR), emaphasizing the usefulness of cryopreservation techniques for plant conservation using Hill’s thistle (Cirsium hillii. (Canby) Fernald). A cryopreservation protocol was established using the droplet-vitrification method for 5-week-old shoot tips of in vitro grown cultures. More than 90% of shoot tips showed regrowth and nearly all regenerated plants were able to survive in the greenhouse. The survival, growth, and development of plants from cryopreserved shoot buds and their performance in field conditions were all comparable or better than the plants from non-cryopreserved buds. Reintroduced plants flowered following overwintering and the magnitude of flowering was site dependent with ca. 80% flowering observed in one site. The site dependent flowering patterns were assessed using phytohormone profiling and compared to herbivory, a common biotic stressor of these plants. Lower tryptophan concentrations led to higher flowering except in alvars, where the limestone resisted root penetration resulting in poor flowering. The presence of tryptamine in the greenhouse acclimatized or alvar field leaves suggested the preparedness of the plants for herbivory/grazing. Serotonin and melatonin concentrations were lower in flowering plants and in sites where the biotic/abiotic stress was minimal. This study provides evidence of the effectiveness of the CPR model in species recovery programs for endangered species. Physiological characterization of plants developed from cryopreserved tissues can be useful for fundamental and applied research in stress adaptation and reproductive biology of plants.

     

Cloning and expression studies on glutathione S-transferase like-gene in honey bee for its role in oxidative stress

Glutathione S-transferases (GSTs) constitute an important multifunctional enzyme family that plays vital roles in cellular detoxification and protecting organisms against oxidative stress caused by reactive oxygen species (ROS). In this study, we isolated a GST-like gene from Apis cerana cerana (AccGSTL) and investigated its antioxidant functions under stress conditions. We found that AccGSTL belongs to the Sigma class of GSTs. Real-time quantitative PCR and western blotting analyses showed that the mRNA and protein levels of AccGSTL were altered in response to oxidative stress caused by various external stimuli. In addition, a heterologous expression analysis showed that AccGSTL overexpression in Escherichia coli (E. coli) cells enhanced resistance to oxidative stress. After AccGSTL silencing with RNA interference (RNAi) technology, the expression of some antioxidant genes was inhibited, and the enzymatic activities of POD, CAT, and SOD were decreased. In conclusion, these data suggest that AccGSTL may be involved in antioxidant defense under adverse conditions in A. cerana cerana.Supplementary InformationThe online version contains supplementary material available at 10.1007/s12192-022-01255-3.     

钦州湾秋季和春季浮游动物分布特征及影响因素

为了解钦州湾浮游动物群落的时空分布特征及与主要环境因子的关系,于2014年10月和2015年3月进行了秋季和春季两航次的调查。结果表明:该海湾的浮游动物群落有明显的季节变化。秋季共鉴定出12类87种,其中优势种有太平洋纺锤水蚤(Acartia pacifica)、肥胖三角溞(Evadne tergestina)、亨生莹虾(Lucifer hanseni)、百陶箭虫(Sagitta bedoti)和长尾类幼虫(Macrura larvae);春季共鉴定出11类48种,优势种为中华哲水蚤(Calanus sinicus)和太平洋纺锤水蚤;秋季浮游动物的平均丰度、生物量和多样性指数(528.92个/m~3、110.60 mg/m~3和2.22)均高于春季(48.30个/m~3、61.10 mg/m~3和1.70)。空间分布上,钦州湾外湾浮游动物丰度、生物量和多样性指数的平均值皆高于内湾。多维尺度分析表明,秋季内湾群落相似性较高,春季外湾浮游动物群落相似性较高。相关性分析表明盐度与营养盐是影响钦州湾浮游动物分布的主要环境因子。与2011—2012年数据相比,钦州湾浮游动物群落结构已趋于单一化和小型化,以致生物量明显下降。这一现象主要与钦州湾海水富营养化以及大面积高密度牡蛎养殖有密切的关系。     

基因编辑动物模型在人类疾病研究中的应用

近年来,随着以CRISPR/Cas9为代表的多种CRISPR系统的开发和不断改进,基因编辑技术逐渐完善,并广泛应用于人类疾病动物模型的制备。基因编辑动物模型为人类疾病的发病机理、病理过程以及预防和治疗等方面的研究提供了重要的素材。目前,用于人类疾病研究的基因编辑动物模型主要有小鼠、大鼠为代表的啮齿类动物模型和以猪为代表的大动物模型。其中啮齿类动物在机体各方面与人类差别较大,且寿命短,无法对人类疾病的研究和治疗提供有效评估和长期追踪;而猪在生理学、解剖学、营养学和遗传学等各方面与人类更接近,是器官移植和人类疾病研究领域重要的动物模型。文中主要介绍了基因编辑动物模型在神经退行性疾病、肥厚心肌病、癌症、免疫缺陷类疾病和代谢性疾病等5种人类疾病研究中的应用情况,以期为人类疾病研究及相关动物模型的制备提供参考。     

基于光合色素的钦州湾平水期浮游植物群落结构研究

应用反相高效液相色谱(RP - HPLC) 并结合二极管阵列检测器分析技术,分析了2010年平水期钦州湾浮游植物光合色素组成,进而由CHEMTAX 软件估算全粒级浮游植物的群落结构。结果表明：平水期浮游植物特征光合色素含量以岩藻黄素最高,其次为叶绿素b、玉米黄素和青绿素,其它特征光合色素含量较低;水体中色素组成表明浮游植物的优势类群为硅藻,其次为蓝藻和青绿藻,它们分别平均占据了浮游植物生物量的73.9%、11.7%和8.7%,其它藻类所占比例很低。硅藻呈现出从湾顶往外随着盐度增加而增加的趋势,而青绿藻显现出相反的分布趋势。钦州湾浮游植物群落结构形成了茅岭江和钦江河口、湾颈-外湾近岸和外湾靠外海域共三种类型,平水期浮游植物群落结构的组成和分布特征主要是由茅岭江和钦江径流变化以及营养盐等环境所决定。平水期叶绿素b和青绿素在大部分站点存在证实了青绿藻在钦州湾的分布,而且表明青绿藻、绿藻和定鞭金藻等微型藻类在钦州湾占有相当比重,它们的重要性有待进一步研究。     

利用响应面方法优化转小鼠金属硫蛋白-I基因聚球藻7002的培养基成分

为实现转小鼠金属硫蛋白基因-I聚球藻7002的高密度培养, 并将其应用于实际的重金属废水处理过程, 首先需要对培养基的成分进行优化。本文利用响应面这一多因素过程优化的有效工具, 通过全因子实验、最陡爬坡实验和中心组合实验, 对转小鼠金属硫蛋白基因-I聚球藻7002培养基的主要成分以及初始pH进行了优化。优化后的培养基组成为: NaHCO3 1.696 g/L, NaNO3 8.57 g/L, 初始pH为8.57, 其他成分同Medium A。优化条件分别在2 L和20 L气升式光生物反应器中得到了验证, 最大细胞浓度分别达到每升4.16 g干重和每升3.12 g干重, 分别比优化前提高了9倍和7倍, 从而为其产业化应用打下基础。     

重组GST-HBx蛋白在表达过程中断裂状况的研究

目的 在对乙型肝炎病毒非结构蛋白HBx研究的过程中,发现体外表达HBx蛋白产物的稳定性存在差异.为了了解其中的原因,拟通过原核表达方式进行分析.方法 以包含全长HBV DNA的质粒pEco63为模板,通过PCR反应,扩增HBV X基因全长序列（1～154氨基酸）和截短序列（48～104氨基酸,48～146氨基酸,98～146氨基酸）,将这几种不同长度和位置的HBX DNA序列克隆到pGEX4T-2原核表达载体中,并通过下游引物设计中引入6×组氨酸标签序列.结果 最终成功构建N端融合GST蛋白标签序列和C端融合6×组氨酸标签序列的4种HBx重组表达载体.将4种包含不同长度HBx的重组质粒转化大肠埃希菌Rosetta,培养至菌液浓度A值大约0.6左右,加入终浓度0.1 mmol/L的IPTG进行诱导表达.提取全菌蛋白通过Western印迹对HBx各重组蛋白产物的N端和C端进行检测.结论 在分段表达的HBx重组蛋白中发生了多处局部的断裂,而断裂部位主要集中于GST蛋白上.而造成这一结果的原因可能与HBx蛋白功能区域暴露后与GST相互作用有关.     

人类3-磷酸甘油醛脱氢酶的原核表达、纯化和鉴定

为了研究3-磷酸甘油醛脱氧酶(GAPDH)化生物学功能,以pcDNA3.1-GAPDH质粒为模板,PCR方法扩增GAPDH基因,经BamHI和SalI双酶切后插入相同酶切的pET32a(+)载体,构建His-GAPDH融合蛋白表达质粒pET32a(+)-GAPDH;转化JM109感受态细胞,并进行阳性克隆筛选,扩增目的质粒;转化大肠杆菌BL21菌株,经IPTG诱导产生融合蛋白,亲和层析柱纯化后,用SDS-PAGE和Western blotting检测GAPDH蛋白表达情况.结果表明,构建的人GAPDH基因表达载体经序列测定证实,与GenBank数据完全一致;双酶切鉴定证实,克隆基因正确插入pET32a(+)载体;表达质粒pET32a(+)-GAPDH在大肠杆菌BL21中成功地诱导表达了可溶性His-GAPDH融合蛋白,纯化后SDS-PAGE和Western blotting检测证实融合蛋白表达成功.人GAPDH原核表达载体的成功构建,及6His-GAPDH融合蛋白的正确表达,为进一步深入研究GAPDH的生物学功能奠定了基础.