A simple, rapid and effective method for total RNA extraction from Lentinula edodes

A rapid, inexpensive and reliable method for total RNA extraction from fruiting bodies of Lentinula edodes containing large quantities of polysaccharides and secondary metabolites is described. An initial extraction step using saturated NaCl solution facilitates the separation of nucleic acids from contaminants and, after further extraction with organic solvents and precipitation with 2-propanol, total RNA of high purity and suitable for applications such as cDNA synthesis, RT-PCR and Northern blot hybridization was obtained. The procedure may also have wider applicability for total RNA extraction from the tissues of other mushrooms.     

短时低温对马铃薯甲虫种群增长的影响

[目的]马铃薯甲虫Leptinotarsa decemlineata是我国重要的检疫性害虫,对茄科植物危害严重.本研究旨在明确出现倒春寒短时低温对马铃薯甲虫种群增长的影响.[方法]马铃薯甲虫卵在8℃下分别处理1,3和5d,以27℃下饲养的卵作为对照,调查卵孵化率及孵化后幼虫的生长发育和成虫繁殖情况,用种群参数评估短时低...     

扶桑绵粉蚧组织蛋白酶B基因的克隆、原核表达和不同发育阶段表达分析

昆虫组织蛋白酶B在昆虫代谢过程中发挥重要作用。本研究利用RACE技术克隆了扶桑绵粉蚧Phenacoccus solenopsis Tinsley组织蛋白酶B基因的开放阅读框（ORF）序列, 命名为PsCb （GenBank登录号: JQ727999）。生物信息学分析表明, 该基因的开放阅读框包含927 bp的片段, 编码308个氨基酸。多序列比对表明, 该基因编码的蛋白在N端变异较大, 在C端保守性高。组织蛋白酶B基因的系统进化树结果表明扶桑绵粉蚧组织蛋白酶独自成为一支。原核表达电泳检测到一条大约35 kDa的目的条带, 与预测的蛋白分子量相符。组织蛋白酶B基因在扶桑绵粉蚧各个虫态均有表达, 卵期表达量相对较低, 2龄若虫期达到最高峰, 然后下降。本研究为进一步研究该基因的功能并开发出组织蛋白酶抑制剂, 从而研制出扶桑绵粉蚧杀卵剂和胚胎发育抑制剂等提供理论依据。     

Endoplasmic reticulum stress and its regulator XBP-1 contributes to dendritic cell maturation and activation induced by high mobility group box-1 protein

High mobility group box-1 protein (HMGB1) had been proved to induce maturation and activation of dendritic cell (DC), however, the endogenous changes and mechanisms underlying are unknown. Since endoplasmic reticulum stress (ERS) activates an adaptive unfolded protein response (UPR) that facilitates cellular survival and repair, we hypothesized that HMGB1 may regulate the function of DC by modulating ERS. In our study, HMGB1 stimulation induced significant ERS responses in DCs in a time- and dose-dependent manner, demonstrated by the up-regulation of a number of ERS markers. Gene silence of XBP-1 in splenic DCs decreased the levels of CD80, CD86 as well as major histocompatibility complex (MHC)-II expression and cytokine secretion after HMGB1 treatment, when compared with untransfected or nontargeting-transfected DCs (all P<0.05). Moreover, XBP-1 silenced DCs after treatment with HMGB1 failed to stimulate notable proliferation and differentiation of T cells, unlike normal DCs or nontargeting-transfected DCs (all P<0.05). Gene silence of XBP-1 resulted in down-regulation of the receptor for advanced glycation end products (RAGE) expression on the surface of splenic DCs induced by HMGB1 stimulation (P<0.05). These findings demonstrate an important role for ERS and its regulator XBP-1 in HMGB1-induced maturation and activation of DCs.     

Up-regulation of mitofusin-2 protects CD4+ T cells from HMGB1-mediated immune dysfunction partly through Ca(2+)-NFAT signaling pathway

High mobility group box 1 protein (HMGB1) was recently discovered to be a critical late-acting cytokine and innate immune-modulating factor in sepsis, but the potential role and mechanism of HMGB1 in adaptive immunity remains elusive. The present study demonstrated that HMGB1 had a dual influence on immune function of CD4(+) T lymphocytes. Low dose of HMGB1 had no effect on the proliferation activity of CD4(+) T lymphocytes, but the Th1 cytokines production was increased. In contrast, treatment with high amount of HMGB1 suppressed the proliferative response and induced Th2 polarization of CD4(+) T lymphocytes. We found that the expression of mitofusin-2 (Mfn2; also named hyperplasia suppressor gene), a member of the mitofusin family, was decreased in CD4(+) T lymphocytes when stimulated with high dose of HMGB1. Up-regulation of Mfn2 attenuated the suppressive effect of HMGB1 on CD4(+) T lymphocytes, which was associated with profound elevation of intracellular calcium concentration ([Ca(2+)](i)) and nuclear factor of activated T cells (NFAT) activity. These results indicate that HMGB1 have a direct role on adaptive immunity, and the decrease of Mfn2 expression may be a major cause of HMGB1-mediated immune dysfunction and Ca(2+)-NFAT signaling defect of CD4(+) T lymphocytes.     

螺旋粉虱产卵蜡泌物对香蕉网蝽的拒食活性

螺旋粉虱产卵分泌物所圈定的叶片范围可阻碍一些植食性昆虫如香蕉网蝽的取食。本文采用叶碟法测定了螺旋粉虱产卵分泌物中的芳香酯类、酚类和烷烃类物质对香蕉网蝽5龄若虫的拒食活性。结果表明:在10mg·mL-1质量浓度下,邻苯二甲酸二丁酯表现出较强的拒食活性,其次为3,5-二(1,1-二甲基乙基)-4-羟基苯丙酸甲酯、邻苯二甲酸正辛酯和正十六烷,4,4’-亚甲基双(2,6-二叔丁基苯酚)则没有拒食活性;浓度梯度试验显示,邻苯二甲酸二丁酯对香蕉网蝽若虫的拒食活性随浓度增大而增强,拒食中浓度为0.929mg·mL-1;表明螺旋粉虱产卵分泌物中的芳香酯对一些植食性昆虫有很强的拒食活性,同时为植食性昆虫种间竞争关系提供了理论依据。     

广东莲花山白盆珠省级自然保护区鸟类资源调查

于2008年1月～2010年12月对广东莲花山白盆珠省级自然保护区鸟类进行了调查,共记录到鸟类151种,隶属于45科16目。其中非雀形目鸟类74种(占49.01%),雀形目77种(50.99%);94种属东洋界种类(62.25%),42种属古北界种类(27.81%),15种属广布种(9.93%);留鸟88种(58.28%)、冬候鸟44种(29.14%)、夏候鸟15种(9.93%)、旅鸟和迷鸟4种(2.65%);国家Ⅱ级重点保护鸟类22种(14.57%)。该保护区鸟类特色是中华秋沙鸭种群数量较大,猛禽资源和水鸟资源较丰富。     

云南省新平县农业生物资源调查与分析

对云南省新平县3个乡镇的11个村进行了重点调查,实地走访了15个村小组,访问了90户农户,共收集到与彝族、傣族、哈尼族、拉祜族生产、生活密切相关的特优、特有、特用农业生物资源164份。分析了当地农业生物资源现状和消长情况及原因和调查、收集到的资源种类及其利用价值,并针对该地区的农业生物资源的特性,提出了利用、保护和开发建议。     

鼠尾胶原蛋白提取、分离、纯化方法的建立及鉴定

目的建立一种高效提取、分离、纯化鼠尾胶原蛋白的方法。方法通过对鼠尾进行剥离获得鼠尾腱,用Tris-HCl缓冲液、胃蛋白酶处理获得鼠尾胶原蛋白原液、反复使用氯化钠溶液进行分级盐析、醋酸溶液复溶进行鼠尾胶原蛋白的纯化。超纯水透析除去无机盐类获得纯化的鼠尾胶原蛋白。通过SDS-PAGE蛋白质电泳、氨基酸含量分析等技术手段鉴定。结果本研究建立的方法可以获得高纯度的鼠尾胶原蛋白,纯度达到电泳纯。与国外进口的商业化鼠尾胶原蛋白产品相比无差异。研究了提取、分离、纯化参数对得率、纯度的影响,建立了最优的鼠尾胶原蛋白提取条件,胃蛋白酶用量：1∶500,酶解时间：72 h,盐析浓度：2 mol/L,提取所用酸溶液：0.05mol/L醋酸溶液。结论为鼠尾胶原蛋白的扩大化生产提供了合适的工艺参数,为大量获得鼠尾胶原蛋白并进行更深层次的功效方面研究提供了理论支持和实践基础。