Changes in leaf ultrastructure and carbohydrates inArabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimation

Summary We studied cell ultrastructure and carbohydrate levels in the leaf tissue ofArabidopsis thaliana L. (Heyn) cv. Columbia during rapid cold acclimation. Freezing tolerance of the leaves from 26 day old plants was determined after 48 h and 10 days at 4°C. Acclimation treatment of 48 h decreased the lethal freezing temperature from –5.7°C to –9.4°C. Freezing tolerance was not altered further by acclimation at 4 °C for 10 days. Ultrastructural changes in the parenchyma cells were evident after 6 to 24 h of cold acclimation. The plasma membrane showed signs of extensive turnover. Evidence of membrane invaginations and sequestering of membrane material was observed. In addition, numerous microvesicles, paramural bodies, and fragments of endoplasmic reticulum were noticed in the vicinity of plasma membrane. Modifications in the structure of cell membranes were evident after 5 days of exposure to low temperature. Small, darkly stained globules were seen on the plasma membrane, tonoplast, chloroplast envelope membrane, mitochondrion outer membrane, dictyosome cisternae membrane, and microvesicle membrane. As far as we are aware, this type of membrane modification has not been described previously in plant cells exposed to low temperature. We propose to call these structures membraglobuli. Acclimation treatment also increased the concentrations of soluble sugars and starch. These observations suggest that cold acclimation inA. thaliana induces changes in both plasma membrane properties and carbohydrate composition.     

凋亡和周期基因芯片研究As2O3对NB4细胞凋亡相关基因表达谱的影响

目的:利用细胞凋亡和周期基因芯片研究As₂O₃作用前后NB4细胞基因表达谱的差异性,寻找As₂O₃诱导NB4细胞凋亡的相关基因并分析其可能机制。方法:流式细胞仪检测细胞凋亡率,抽提对照及诱导组细胞的mRNA,通过逆转录将As₂O₃处理前后的NB4细胞cDNA进行生物素标记,用含269个目的基因的细胞凋亡和周期基因芯片进行杂交,GEArray软件分析,筛选出As₂O₃诱导前后表达有差异的基因。芯片结果用荧光定量聚合酶链反应(realtimepolymerasechainreaction)进行验证。结果:筛选出As₂O₃作用前后表达有差异的基因共100条(占芯片基因总数的37.2%),其中97条(97/100,97%)基因表达上调,3条(3/100,3%)基因表达下调。表达上调的基因主要包括肿瘤坏死因子配体和受体家族、bcl2家族、半胱氨酸家族、DNA损伤检测和P53途径以及细胞分裂周期蛋白和激酶等基因。结论:As₂O₃主要通过上调促凋亡基因表达来诱导NB4细胞凋亡,其中TNFSF15、Apaf1、Caspase3和p16等基因可能参与As₂O₃诱导的NB4细胞凋亡,As₂O₃诱导NB4细胞凋亡的可能机制主要涉及TNF途径、线粒体途径、Caspase途径、细胞周期抑制途径和P53途径等。     

An improved definition of mouse mammary epithelial side population cells

BACKGROUND: Mammary epithelial side population cells have been suggested as candidate mammary stem cells. To date, for technical reasons, these cells have been poorly defined and cross-comparison of data between different laboratories has been difficult. Here, we set out to define mammary side population cells in a way that improves the ability to carry out such comparisons. METHODS: Mouse mammary epithelial cells were stained with Hoechst 33342. Light scatter, PI staining and clonogenicity of different regions of the Hoechst profile were examined. Time-course analyzes of Hoechst 33342 loading were carried out. RESULTS: Detailed examination of the light scatter and PI staining of Hoechst 33342-stained mammary cells enabled single live side population and non-side population cells to be defined with greater accuracy. Comparison of ABC pump inhibitors identified potential discrepancies in results obtained using these inhibitors. Time-course analyzes enabled side populations cells to be identified as a dynamic cell population that could be defined accurately by using the relationship between Hoechst 33342-staining profiles of consecutive time points. DISCUSSION: Defining the side population of solid tissues as a 'stabilized side population percentage' will enable a more rigorous study of the side population phenomenon and improve evaluation of results from different laboratories.     

基于GIS和GS的东亚飞蝗卵块空间格局的研究

本文以沿渤海蝗区东亚飞蝗Locusta migratoria manilensis (Meyen) 越冬卵块为研究对象,野外采用450 m和50 m规则栅格取样,包括卵量、植被覆盖度、土壤含水量、含盐量、土壤pH和有机质等,利用地统计学方法,在GIS平台下,分析研究区域内影响蝗虫产卵选择的环境因子、蝗虫卵块的空间异质性及分布格局。结果表明, 植被覆盖度、土壤含盐量和土壤含水量在有卵和无卵的环境中存在极显著的差异,飞蝗产卵时最适宜的植被覆盖度、土壤含水量和盐度范围分别为0～30%、10.1%～20.0%和0.09%～1.99%,且当小环境植被覆盖度>50%、土壤含水量>30%或含盐量>3%时,飞蝗不再选择产卵。蝗虫卵块具有高度空间异质性,其空间自相关范围平均为390 m且呈斑块、聚集分布,蝗虫卵块变异函数曲线为球状模型。利用块段克立格法进行空间局部插值,得到研究区域卵块的空间分布格局图,可较准确地描述飞蝗卵块在研究区域内的空间分布、形状、地理位置及相对位置。研究结果可为地面卵块抽样调查、实时跟踪蝗卵胚胎发育进程、确定蝗灾早期发生点、片防治区域及蝗灾早期遥感预警提供科学依据。     

利用重组荧光素酶报告基因载体研究E2F调控组蛋白H2A的转录

转录因子E2F与细胞增殖、凋亡及癌变密切相关,组蛋白H2A是构成核小体的重要成员之一.研究通过特异性的阻断Rb基因的表达发现组蛋白H2A家族成员:HIST1H2AJ表达下调,再进一步研究转录因子E2F对HIST1H2AJ的转录调控.通过PCR得到HIST1H2hJ的5非翻译区序列,克隆到荧光素酶报告载体pGL3中,然后转染入HEK293,再检测荧光素酶的活性来判断E2F是否调控HIST1H2AJ的转录.研究结果表明:转录因子E2F能够调控组蛋白H2A家族成员之一HIST1H2AJ的转录.     

热环境对虎斑颈槽蛇卵孵化期、孵化成功率和孵出幼体特征的影响

用恒温（２４、２７、３０和３３Ｃ）和波动温度（平均２６．１Ｃ,范围２０．１—３２．７Ｃ）孵化虎斑颈槽蛇（Ｒｈａｂｄｏｐｈｉｓ　ｔｉｇｒｉｎｕｓ　ｌａｔｅｒａｌｉｓ）卵,检测热环境对孵化期、孵化成功率和孵出幼体的影响。孵化热环境显著影响孵化期、孵化成功率和胚胎畸形率,对孵出幼体性别无显著影响。孵化期随孵化温度升高而缩短,２４、２７、３０和３３Ｃ的平均孵化期分别为４５．０、３２．７、２７．３和２６．０ｄ,波动温度的平均孵化期为３７．９ｄ。３３Ｃ孵化成功率最低（１６．７％）,胚胎畸形率最高（１００％）。孵出幼体总性比（雌性／雄性＝０．６）不显著偏离１：１。孵出幼体的尾长显示两性异形,雄性尾长大于雌性;其它被检幼体特征无显著的两性差异。２４、２７和３０Ｃ以及波动温度孵出幼体的所有被检指标均无显著差异。３３Ｃ孵出幼体的体重和个体大小小于其它热环境中孵出的幼体,并特征性地具有较小的躯干、较大的剩余卵黄。３３Ｃ中胚胎发育的能耗显著大于其它热环境中胚胎发育的能耗。３３（‘孵出幼体的灰分含量较低,但孵出卵卵壳较重。３３Ｃ孵出幼体不能运动;其它热环境中孵出的幼体在跑道上表现良好,这些幼体的不问断运动的最大距离、每分钟运动距离和每分钟停顿次数无显著的差异。结果表明,持续将虎斑颈槽蛇卵暴露在３３Ｃ条件下不利于该种胚胎发育并可能对胚胎具有致死性影响,波动温度孵卵有利于拓宽存活孵化温度范围。     

Promotive Effect of Low Concentrations of NaHSO3 on Photophosphorylation and Photosynthesis in Phosphoenolpyruvate Carboxylase Transgenic Rice Leaves

Spraying a 1-2 mmol/L solution of NaHSO3 on the leaves of wild-type rice (Oryza sativa L.)Kitaake (WT), phosphoenolpyruvate carboxylase (PEPC) transgenic (PC) rice and PEPC phosphate dikinase(PPDK) transgenic rice (PC PK), in which the germplasm was transformed with wild-type Kitaake as the gene receptor, resulted in an enhancement of the net photosynthetic rate by 23.0%, 28.8%, and 34.4%,respectively, for more than 3 d. It was also observed that NaHSO3 application caused an increase in the ATP content in leaves. Spraying PMS (a cofactor catalysing the photophosphorylation cycle) and NaHSO3 separately or together on leaves resulted in an increase in photosynthesis with all treatments. There was no additional effect on photosynthetic rate when the mixture was applied, suggesting that the mechanism by which NaHSO3 promotes photosynthesis is similar to the mechanism by which PMS acts and that both of compounds enhanced the supply of ATE After spraying a solution of NaHSO3 on leaves, compared with the WT Kitaake rice, a greater enhancement of net photosynthetic rate was observed in PEPC transgenic(PC) and PEPC PPDK transgenic (PC PK) rice, with the greatest increase being observed in the latter group. Therefore ATP supply may become the limiting factor that concentrates CO2 in rice leaves transformed with an exogenous PEPC gene and exogenous PEPC PPDK genes.     

30 nm染色质的体外组装和电镜分析

真核生物的基因组以染色质的形式存在,染色质在真核生物的基因表达调控及胚胎发育过程中起重要作用,为表观遗传提供一个重要的信息整合平台．染色质的高级结构,特别是 30 nm染色质的动态变化在基因转录沉默和激活过程中起着重要的调控功能．但是目前对30 nm 染色质纤维的组装及其精细结构的认识还十分有限．本文通过体外表达系统,表达未经修饰的组蛋白,并利用克隆构建的601DNA均一重复序列,通过逐步降低盐离子浓度并加入组蛋白H1或镁离子的方法,体外重组均一的30 nm染色质纤维．并利用镀金属、负染色制样和冷冻电镜制样等手段通过透射式电子显微镜(TEM)对30 nm纤维结构的形成原因、组蛋白H1的作用和核小体重复单位(nucleosome repeat lengths,NRLs)长度对30 nm染色质纤维的影响进行研究．研究结果显示在组蛋白H1或二价镁离子存在的情况下,均可形成30 nm染色质纤维．其形成的染色质拓扑结构有所不同．统计分析表明,不同长度核小体重复单位(NRLs)形成的染色质纤维直径有所不同(P < 0.05)．同时,我们得到了较为均一的冷冻电镜样品,为进一步研究30 nm染色质纤维的高级结构及理解体内染色质存在的形式及动态过程打下了较好的基础．     

响应面分析法优化当归粗多糖提取工艺

选取岷县当归药材为原料,采用水提醇沉法进行多糖提取,以当归粗多糖得率为指标,探讨加水量、回流提取时间、水提液的浓缩比、醇沉后所达含醇比例对当归多糖得率的影响。在单因素分析的基础上,采用响应面分析法(RSM)确定了当归粗多糖最佳提取条件为:加水量837.6 mL/100 g,浓缩后溶液体积为228.12 mL,最终含乙醇浓度为65.80%,回流提取时间2 h,在此条件下预测当归粗多糖得率理论的最佳值为10.44%,实际验证值为10.40%,两者相符,说明RSM法分析的可靠性。