拟南芥U-box蛋白的研究进展

泛素-26s蛋白酶体途径（ubiquitin-26s proteosome pathway,UPP）是真核生物体内一种高度选择性的蛋白降解途径。U—box蛋白是该途径中决定底物特异性的一种泛素连接酶E3。U—box结构域大约由70个氨基酸残基构成,在酵母、植物和动物等真核生物中高度保守。植物U-box蛋白（plant U—box protein,PUB）的数目远多于其他生物,在植物的生长发育过程中起着重要作用。目前在拟南芥中已发现60多个PUB蛋白（Arabidopsis plant U-box protein,AtPUB）,成为近年来受到较多关注的一类蛋白。本文根据蛋白的结构特征和系统进化树比较,将61种AtPUB蛋白分为7大类。同时,对已报道AtPUB蛋白的功能进行了综述并提出了研究展望。     

Dihydroartemisinin enhances radiotherapy efficiency in H22 cell tumor-bearing mice by regulating PI3K/AKT signaling pathway

AIM To study the effect of dihydroartemisinin （DHA） on the radiotherapy efficiency in hepatocellular carcinoma H22 cell tumor-bearing mice and the role of phosphatidylinositol 3-kinase （PI3K）/protein kinase B （AKT） signaling pathway in this process. METHODS A model of H22 cell tumor-bearing mice was established. The mice was divided into model group, single radiotherapy group, 5-fluorouracil （5-FU） group, and low-, medium- and high-dose DHA groups. The body weight and tumor volume in each group were measured every other day. At the end of administration, blood was collected from the tail of the mice and the animals were killed by neck removal immediately. The synergistic effect of DHA on radiotherapy was determined, and tumor growth inhibitory rate was calculated. The degree of lymphocyte transformation and natural killer （NK） cell activity were measured by MTT, the serum levels of interleukin-2 （IL-2） and IL-4 were measured by ELISA, and the protein levels of PI3K, AKT and p-AKT were determined by Western blot. RESULTS The H22 cell tumor-bearing mouse model was successfully constructed. Compared with model group, the TGT₃ （tumor growth time to reach 3 times of volume） of single radiotherapy group was remarkably increased （P<0.05）, while tumor weight, lymphocyte transformation degree, NK cell activity, IL-2 and IL-4 levels, PI3K protein level and AKT phosphorylation level were remarkably decreased （P<0.05）. Compared with single radiotherapy group, TGT₃, EF （enhancement factor）, tumor inhibitory rate, lymphocyte transformation degree, NK cell activity, IL-2 level and IL-4 level were increased with the increase in DHA dose （P<0.05）, and the PI3K protein level and AKT phosphorylation level were decreased （P<0.05）. CONCLUSION DHA may enhance the immunity of tumor-bearing mice by inhibiting the activity of PI3K/AKT signaling pathway, thereby enhancing the efficacy of radiotherapy.     

辣椒新品种平椒3号的选育

平椒3号是以自交系99101为母本,自交系9949为父本配制的早熟一代杂种。果实羊角形,青熟果深绿色,果纵径17.7cm,果肩径3.0cm,果肉厚0.25cm,平均单果质量29.5g,肉质脆嫩,辣味浓且有香味,商品性好。田间对疫病、炭疽病、病毒病的抗性明显优于猪大肠。露地栽培每667m2青椒产量为3700kg左右,适宜西北地区及四川、山西、湖南等嗜辣地区露地栽培。     

2,4-表油菜素内酯和生长素促进大麦胚芽鞘伸长生长的协同作用

以大麦胚芽鞘为材料,研究了生长素吲哚乙酸(indole-3-acetic ac id,IAA)和2,4表油菜素内酯(2,4-ep ibras-sinolide,ep iBR)对胚芽鞘伸长的影响。结果表明:200μmol.L-1IAA和1μmol.L-1ep iBR在黑暗条件下均促进大麦胚芽鞘的伸长生长,而且在该过程中二者存在协同作用,但必须是在IAA之前施用ep iBR或两者共同处理时才能表现;100 mg.L-12,3,5三碘苯甲酸(2,3,5-triiodobenzoic ac id,TIBA)能抑制ep iBR诱导的胚芽鞘伸长生长,暗示IAA的极性运输是ep iBR促进大麦胚芽鞘伸长生长所必需的;内源激素分析表明,ep iBR处理后胚芽鞘内源IAA含量上升,暗示ep iBR对大麦胚芽鞘伸长的促进作用在一定程度上通过调控内源IAA的水平起作用。     

牦牛MYL3基因的克隆及组织表达分析

【目的】研究牦牛MYL3基因的结构和表达特征,探究其生物学功能及影响牦牛肌肉生长发育的机制。【方法】以美仁牦牛cDNA为模板,PCR扩增MYL3基因编码区序列(CDS),利用MEGA11软件构建系统进化树,利用生物信息学软件对其编码蛋白进行分析。通过实时荧光定量PCR(RT-qPCR)检测MYL3基因在心脏、肝脏、脾脏、肺脏（参照）、睾丸、肌肉和脂肪组织中的相对表达量。【结果】牦牛MYL3基因CDS区长600 bp,共编码199个氨基酸,存在1个碱基突变位点,位于第165位(A>T)。系统进化分析结果显示,牦牛与普通牛的亲缘关系最近,与单峰驼的亲缘关系最远。生物信息学分析结果显示,牦牛MYL3蛋白为不稳定的亲水性蛋白,无信号肽,不存在跨膜结构,主要存在于细胞质内,有8个磷酸化位点和2个N 糖基化位点,蛋白的二级结构以α-螺旋为主,主要与调控肌肉生长发育的相关蛋白相互作用。RT-qPCR结果表明,MYL3基因在心脏中的表达量最高,极显著高于其他组织;肌肉中次之,与除心脏外的其他组织存在极显著差异。【结论】MYL3基因可能与牦牛心脏发育有关,对肌肉的生长发育和肉质有一定影响。     

苏云金芽孢杆菌营养期杀虫蛋白Vips研究进展

苏云金芽孢杆菌Bt营养期杀虫蛋白Vips主要分为Vip1、Vip2和Vip33种。Vip1和Vip2构成二元毒素,对叶甲科昆虫具有特异杀虫活性。Vip3对鳞翅目昆虫具有广谱杀虫活性,该蛋白广泛存在于Bt中,与已知杀虫晶体蛋白ICPs没有序列相似性。Vip3通过诱发细胞凋亡,最终导致昆虫死亡,这与Btδ-内毒素的作用机理完全不同,Vips的发现对于Bt的进一步开发利用具有重要意义。本文主要对营养期杀虫蛋白Vips的分布、特性、作用机制及应用等进行报道。     

果蔗脱毒种苗组培快繁技术研究

采用不同浓度的6-BA对果蔗进行了茎尖分生组织的诱导和幼芽增殖试验研究,利用不同状态的培养基对增殖芽进行了培养。结果表明,当6-BA浓度为4-5mg／L时,比较适宜于果蔗茎尖分生组织的诱导。在幼芽增殖前期．6-BA的浓度采用2-3mg／L;在将进入生根阶段时,6-BA的浓度采用0．5-1．5mg／L,有利于获得粗壮的幼苗。果蔗幼芽在增殖阶段。适宜采用液体培养基,可降低成本。     

猕猴桃AdCCS1基因的克隆及其表达调控

采用RT-PCR法从‘米良1号’猕猴桃中分离到1 066 bp的铜锌超氧化物歧化酶分子伴侣蛋白基因AdCCS1,登录号为KY471361。AdCCS1的开放阅读框为984 bp,编码327个氨基酸,包含3个典型结构域（N端结构域、中间结构域和C端结构域）和2个保守的金属结合位点（MXCXXC和CXC）。基因结构分析显示AdCCS1由6个外显子和5个内含子组成。系统进化分析表明AdCCS1聚在双子叶植物分支中,与茶树CsCCS的亲缘关系最近。此外,AdCCS1的5′端调控区存在多种光响应、胁迫响应和激素响应应答元件。定量PCR分析显示,AdCCS1在猕猴桃叶片中的表达量最高,其次是成熟果、花、幼果和茎,在根中的表达量最低。猕猴桃果实中AdCCS1在4 ℃低温贮藏过程中的表达量均比25 ℃贮藏中同期的低,说明低温抑制AdCCS1的表达。脱落酸和赤霉素处理后AdCCS1的表达下调,但二者的应答模式不同。     

Effects of baicalin on CA46 cell xenografts in nude mice

AIM: To study the effects of baicalin on CA46 cell xenografts in nude mice. METHODS: The nude mice with CA46 cell xenografts were treated with drugs via intraperitoneal injection daily, and were divided into 5 groups: negative control group, 15 mg/kg baicalin group, 30 mg/kg baicalin group, 60 mg/kg baicalin group and 4 mg/kg etoposide (VP-16) positive control group. After 12-day treatment, the weight of CA46 cell xenografts stripped from some nude mice in the 5 groups was used to evaluate the effect of baicalin on xenograft growth in the nude mice. The apoptosis, necrosis and pathological changes of the xenograft cells were examined under light microscope and transmission electronic microscope respectively. The expression levels of phosphatidylinositol 3-kinase (PI3K)/protein kinase B (Akt) signaling pathway-related proteins extracted from xenografts were determined by Western blotting. The other nude mice with CA46 cell xenografts in the 5 groups continued to be treated with the drugs until death in order to evaluate the effect of balcalin on survival time of the nude mice with CA46 cell xenografts. RESULTS: Baicalin remarkably inhibited the growth of CA46 cell xenografts, induced apoptosis and necrosis of xenograft cells, and reduced the protein expression of phospho-Akt (p-Akt), nuclear factor-kappa B (NF-κB), mammalian target of rapamycin (mTOR) and phospho-mTOR (p-mTOR) in the xenografts after 12-day treatment. Furthermore, baicalin prolonged the survival time of the nude mice with CA46 cell xenografts in a dose-dependent manner. CONCLUSION: Baicalin inhibits the growth and induces apoptosis of CA46 cell xenografts in the nude mice, and prolongs the survival time of the nude mice with CA46 cell xenografts through the mechanism of down-regulating PI3K/Akt/NF-κB and PI3K/Akt/ mTOR signaling pathways.     

黄皮素内酯Ⅱ在黄皮植物体的分布及对稗草生化代谢的影响

[目的]测定黄皮素内酯Ⅱ(2′,3′-epoxyanisolactone)在黄皮[Clausena lansium(Lour.)Skeels]植株的花、枝叶和果核中的含量,明确其对稗草生化代谢的影响。[方法]采用液相色谱法进行含量的测定,小杯法培养供试稗草。[结果]黄皮素内酯Ⅱ在花粗提物中的含量较少,仅为0.43%,在枝叶和果核中的含量分别为3.33%和3.21%。黄皮素内酯Ⅱ处理稗草植株,处理后稗草根中蛋白含量和氨基酸含量与未经处理存在明显的差异,在50μg/mL的浓度下,可以显著地降低稗草植株内的蛋白含量和增加植株内的氨基酸含量。当处理浓度为50、25、12.5、6.25、3.125μg/mL时,样品中的含氮量分别为1.402 8、1.317 2、1.250 2、1.128 7、0.903 2mg/g,比对照0.832 1mg/g均有所增加。黄皮素内酯Ⅱ处理后,稗草的蛋白含量与对照相比有所下降,处理浓度为50、25、12.5、6.25、3.125μg/mL时,蛋白含量比对照分别降低75.91%、66.43%、62.43%、58.57%和58.03%。[结论]黄皮素内酯Ⅱ对稗草代谢有一定影响。