不同浓度含氨基酸水溶肥对小果型西瓜根际土壤环境及果实品质的影响

为进一步研究含氨基酸水溶肥对西瓜根际微生物群落的作用,通过大田试验分析不同浓度含氨基酸水溶肥(氨基酸含量(ρ,后同)≥100 g·L-1,钙+镁含量≥30 g·L-1,试验共分4个处理,CK:传统均衡型水溶肥;T1:含氨基酸水溶肥浓度(w,后同)为0.200％;T2:含氨基酸水溶肥浓度为0.125％;T3:含氨基酸水溶...     

瓜类果斑病菌（Acidovorax citrulli）基因组信号肽预测分析

利用Signal 3．0、LipoP和TargetP对瓜类果斑病菌Acidovorax avenae subsp．chrulli AAC00—1菌株基因组中4709个ORFs进行分析．对该病菌基因组中信号肽的数量、长度和氨基酸组成进行了预测,并对其进行分类。结果确定其中476个ORFs所编码的N-端有信号肽序列,占全部ORFs的10．10％。信号肽长度为14—48个氨基酸,以20～35个氨基酸居多．26个氨基酸的信号肽最多。组成信号肽的氨基酸中,非极性氨基酸占47．80％,极性氨基酸占23．04％．带负电荷的酸性氨基酸占15．87％,带正电荷的碱性氨基酸占8．73％。预测的476条信号肽中,有384条分泌型信号肽（SPI）,40条脂蛋白型信号肽（sPII）,51条TM}Ⅱ型信号肽和1条CYT型信号肽。在分泌型信号肽中,有96个信号肽具有RR—motif的保守区段。     

化肥减施条件下土壤改良剂及氨基酸叶面肥对水果黄瓜生长和产量的影响

在化肥减施条件下,为探究生长调节剂及氨基酸叶面肥对水果黄瓜生长及品质的影响,开展了田间试验,共设置5个处理,T1,空白对照;T2,常规施肥;T3,常规施肥+土壤改良剂;T4,土壤改良剂+20％化肥减施;T5,土壤改良剂+20％化肥减施+氨基酸叶面肥.试验结果表明,在化肥减施的情况下,配施土壤改良剂和氨基酸叶面肥的T5处理可以显著提高水果黄瓜产量和生长指标,水果黄瓜产量、株高、茎粗、叶绿素分别提高了39％、19％、20％、18％,T5处理的经济效益较常规施肥有明显的增加.因此,在化肥减施20％条件下,配施氨基酸叶面肥和土壤改良剂可以有效促进水果黄瓜产量和经济效益的增加.     

贝尔格莱德灵芝人工驯化栽培及其化学成分分析

以栎属原木为材料,对来自贝尔格莱德的9个灵芝菌株进行短段木栽培试验,结果发现,GA．2和GA．9两个菌株尚具一定栽培价值,单产分别为8．8kg/m^3和8．3kg／m^3,平均单朵重分别为12．38g和12．16g,GA．2和GA．9两个菌株的聚丙烯酰胺凝胶电泳图谱基本一致。测试结果表明,驯化栽培的灵芝子实体,其水分、粗蛋白、粗纤维、粗脂肪、多糖及灰分含量分别为11．3％～12．7％,11．6％～12．8％,28．7％～30．8％,4．8％～5．6％,0．90％～1．02％和1．4％～1．7％;其脂肪酸构成主要以油酸(c18：l,59．6％)和亚油酸(C18：2,19．2％)等不饱和脂肪酸为主;含有18种氨基酸,总量为9．611％,其必需氨基酸含量占氨基酸含量总量的50．6％(E／T=0．506)。     

不同基料种植的姬松茸营养成分的比较研究

利用原子分光光度法、苯酚一硫酸法、氨基酸分析法及凯氏定氮法,分析比较了以芦笋老茎为主要基料栽培的姬松茸子实体和传统以稻杆为主要基料种植的姬松茸子实体中Ca2＋、多糖、蛋白质和氨基酸的含量。结果表明,芦笋老茎做基料培育的姬松茸中钙含量比普通姬松茸高66．71％,多糖、总蛋白质和总氨基酸的含量分别高22．49％、6．3％、28．04％。     

西瓜抗病基因同源序列的克隆与分析

以植物丝氨酸／苏氨酸蛋白激酶类（STK）抗病基因产物催化结构域l的保守氨基酸序列设计简并引物,以西瓜基因组DNA为模板进行PCR,扩增,得到大约500bp的目的条带。重组质粒经PCPo检测后进行测序,获得1个有效序列,可以编码完整的氨基酸序列。同源性分析表明其具有STK保守结构域,与已经克隆的Pto、LrlO和Lectin等基因在氨基酸水平上的同源性为40．O％～54．O％。     

扁桃SLF基因和S-RNase基因的克隆及表达分析

 以扁桃‘Pioneer’品种为材料, 利用RT-PCR及RACE技术, 克隆了1个新的SLF ( S LocusF-box ) 基因(PdSLF1) 和两个新的S-RNase基因( PdSm 和PdSn) 的cDNA。PdSLF1全长1 331 bp, 编码376个氨基酸; PdSm 全长826 bp, 编码228个氨基酸; PdSn基因全长878 bp, 编码227个氨基酸。与公共数据库中的序列进行相似性比较, 发现这3个基因所编码的氨基酸序列与其它蔷薇科植物相应基因的氨基酸序列均具有较高的一致性, PdSLF1为70.2% ～84.8% , S-RNase为59% ～83.9%。PdSLF1基因在花药中专一性表达, 而PdSm 和PdSn基因在雌蕊中专一性表达。     

猪苓营养菌丝与野生菌核蛋白质成分分析

测定和分析了猪苓(Polyporus umbellatus)营养菌丝与野生菌核中蛋白质的氨基酸种类及含量。结果表明,猪苓营养菌丝中的粗蛋白含量是32．5％,必需氨基酸含量是3．51％,氨基酸总量是10．09％;野生菌核中的粗蛋白含量是6．87％,必需氨基酸含量是1．53％,氨基酸总量是4．53％。营养菌丝的粗蛋白和氨基酸含量显著高于野生菌核。     

秦巴山区野生猪苓菌核的品质分析

采用FDBN柱前衍生高效液相色谱法等分析了秦巴山区野生猪苓茵核的氨基酸、常规营养成分和猪苓多糖含量。结果表明，所测定的17种氨基酸中氨基酸总量为4521．0mg/100g，7种必需氨基酸占氨基酸总量的33．8％；水分、灰分、粗蛋白、粗纤维、粗脂肪、猪苓多糖含量分别是13．22％、2．86％、6．87％、27．62％、1．87％和0．51％。     

胶陀螺营养成分分析

测定了胶陀螺的营养成分，结果为总糖17．7％，脂肪2．1％，蛋白质10．93％；氨基酸总量6．84％，其中必需氨基酸为45．62％，其它氨基酸为54．38％；主要无机元素钙3．87，镁1．1012，铁0．324，磷3．67，锌0．003（单位：g/g)；挥发油的平均收率为0．07％，经鉴定主要成分为邻苯二甲酸丁辛酯，棕榈酸，亚油酸、油酸、环戊烷基十一烷酸。     

萱草叶斑病的病原鉴定及其生物学特性

为明确在贵州发现的萱草（Hemerocallis fulva）叶斑病病原菌及其生物学特性，采用组织分离法和离体接种法分别对其病原菌进行分离和致病性测定，利用形态学及ITS基因序列分析对病原菌进行鉴定并对其生物学特性进行研究。鉴定结果表明：分离得到的萱草叶斑病病原菌菌株（编号为XCS369）的菌丝为灰白色、绒毛状，菌落中心隆起呈灰褐色，背面橄榄绿色，培养15 d表面可产生白色孢子粉；分生孢子光滑，无色，椭圆至长椭圆形。在以ITS基因序列构建的系统发育树中，XCS369与古巴炭角菌（Xylaria cubensis）聚于一支，且支持率达99%，结合形态特征与系统发育分析将其鉴定为古巴炭角菌。该菌菌丝最适生长温度为25 ~ 28 ℃，最适pH 7，在马铃薯葡萄糖培养基上生长最快，对碳源麦芽糖、氮源酵母浸粉的利用最高，菌丝生长对光照不敏感。     

萱草再生体系技术建立的研究

以萱草的根茎为材料,进行愈伤组织诱导和分化,试管苗的生根、移栽和移植的研究,建立萱草再生体系技术。结果证明:MS+BA0.4mg/L+NAA0.1mg/L+2,4-D0.1mg/L是愈伤组织诱导培养的理想培养基;MS+NH_4H_2PO_450mg/L+BA1.0mg/L+NAA0.1mg/L是愈伤组织增殖继代培养的理想培养基;MS+AgNO_30.5mg/L+GA_30.5mg/L+BA0.5mg/L+NAA0.1mg/L是愈伤组织分化培养的理想培养基;1/2MS+NAA0.1mg/L+IAA0.2mg/L是不定芽生根培养的理想培养基。在河沙中试管苗易移栽成活;移植到花坛中的试管苗根系发达、生长旺盛。     

昆明常见园林地被植物耐旱性研究

通过盆栽4种多年生观赏地被植物,对其进行连续20d的自然失水胁迫处理,研究供试植物在干旱胁迫下的土壤相对含水量以及叶片的枯叶率、游离脯氨酸、相对电导率的变化。结果表明:在干旱胁迫下各参试植物的各指标变化显著,综合分析各指标,4种观赏植物抗旱性由强至弱排序为:松叶景天萱草头花蓼紫花地丁。松叶景天、萱草具有很强的抗旱性,在园林绿化中可大力推广应用。     

结球甘蓝春化相关基因VIN3反义植物表达载体构建及转化

以pBI121植物表达载体和BoVIN3-1基因片段为基础,构建结球甘蓝春化相关基因VIN3反义植物表达载体。将BoVIN3-1基因片段反向插入355启动子与GUS基因之间的限制性酶切位点XbaⅠ和SmaⅠ中,构建含反义BoVIN3-1基因的工程质粒pBI35S-BoVIN3-1。通过花蕾微量注射法转化结球甘蓝,获得9株转化植株。PCR检测结果表明,其中5株为阳性植株,阳性株率55.6%。经春化处理的转反义基因植株与对照相比,春化一定程度被推迟。半定量RT-PCR检测结果表明,转化植株在进行低温处理后仍有该基因少量表达,并随春化处理时间的延长转录水平升高,在第50天时表达量达到最高。     

杨梅酸性转化酶基因cDNA分离及表达分析

杨梅果实富含蔗糖,酸性转化酶是蔗糖代谢关键酶,根据植物酸性转化酶基因保守区序列设计引物,提取杨梅叶片RNA,逆转录获得cDNA,以此为模板通过PCR技术扩增到长度为516bp的基因片段,克隆入pMD18-T载体中,命名为MrIVR1(GenBank:DQ339699)。测序及同源性检索表明,该基因推导氨基酸序列与君子兰、葡萄、草莓、胡萝卜等酸性转化酶基因氨基酸序列同源性为60%～69%。运用ClustalX软件对植物转化酶基因进行了系统树分析,结果显示,MrIVR1编码的蛋白质属于细胞壁酸性转化酶。半定量RT-PCR表达分析显示,MrIVR1基因在杨梅果实发育早期表达量最高,随着果实的发育表达量下降,在成熟果实中表达水平较低。     

Conditionally replicating adenovirus activated by CXCR4 promoter in lung cancer

AIM: To construct a conditionally replicating adenovirus vector activated by CXCR4 promoter and to evaluate its ability of lysing the lung cancer cells specifically. METHODS: Human CXCR4-E1A gene amplified by PCR was cloned into the shuttle plasmid pDC316-GFP to construct the recombinant shuttle plasmid pDC316-CXCR4-GFP. The recombinat shuttle plasmid and adenovirus genomic plasmid pBHG-lox-E1, 3Cre were transfected into 293 cells to construct the recombinant adenovirus CRAd-CXCR4-GFP. PCR was used to detect the target gene fragments, and the viral titer was determined. A549 cells with the highest mRNA expression of CXCR4 were screened out from 5 kinds of lung cancer cell lines by real-time PCR. CXCR4 promoter activity and adenovirus replication numbers were detected in A549 cells after transfection of CRAd-CXCR4-GFP and Ad-NULL. CRAd-CXCR4-GFP and Ad-NULL were transfected into A549 cells and 16HBE cells, the apoptotic rates were detected by flow cytometry and the viability was analyzed by CCK-8 assay. RESULTS: The recombinant plasmid pDC316-CXCR4-GFP was constructed successfully. Green fluorescence was observed in 293 cells under fluorescent microscope after co-transfection of pDC316-CXCR4-GFP and pBHG-lox-E1, 3Cre at 11 d. Green fluorescence was observed in 293 cells after infection of amplified 3rd generational adenovirus. PCR showed that the purpose gene was successfully integrated in recombinant adenovirus genome. The virus in the supernatant reached a titer of 1×10¹³ PFU/L. The mRNA expression of E1A and E4 in the A549 cells after transfection of CRAd-CXCR4-GFP was markedly increased compared with Ad-NULL group. Compared with Ad-NULL group and empty control group, the apoptotic rate and the viability of A549 cells in CRAd-CXCR4-GFP group had no significant difference in the first 4 d, the apoptotic rate increased significantly at 5 d, and the cell viability declined significantly at 5 d, but the apoptotic rate and the viability of 16HBE cells in each group had no significant difference within 5 days. CONCLUSION: The conditionally replicating adenovirus vector CRAd-CXCR4-GFP has been successfully constructed, which has the ability of lysing lung cancer cells specifically.     

CREG gene silencing induces spontaneous apoptosis of ESCs-derived Ebs

AIM: To investigate the effect of cellular repressor of E1A-stimulated genes (CREG) on the spontaneous apoptosis of murine embryonic stem cells(ESCs)-derived embryoid bodies (EBs).METHODS: The murine ESCs R1 were infected with pDS-shRNA-CREG and pDS-shRNA-GFP retrovirus, respectively. R1, R1-GFP and R1-shCREG were cultured on STO feeder cells in DMEM supplied with leukemia inhibitory factor(LIF). Alkaline phosphatase(AKP) staining and teratoma formation assay inoculated into mouse myocardium were used to detect stemness of transfected ESCs. R1/EB, R1-GFP/EB and R1-shCREG/EB were produced by liquid suspension method. The expression of CREG and cleaved caspase-3 were analyzed by Western blotting and quantitative RT-PCR on day 7. The apoptotic rates of the 3 kinds of EBs were analyzed by flow cytometry(FCM) analysis with Annexin V/PI dual staining. RESULTS: The stably-transfected ES cells (R1-shCREG and R1-GFP) were obtained by screening the G418-resistant clones. R1-GFP and R1-shCREG had the stem cell properties similar to those of R1 detected by AKP staining. However, it was found that R1-shCREG didn't show the same almighty differentiation function as of R1 and R1-GFP by tumor experiments in mouse myocardium that it couldn't form teratomas analogue and had the lower ability of cell differentiation. Observation under phase-contrast microscope showed that the cell differentiation was inhibited while the number of cell death was increased in R1-shCREG/EB group. Compared with R1/EB and R1-GFP/EB, Western blotting analysis demonstrated that the protein expression of CREG was decreased to (78.0±1.3)% and (84.0±2.4)% on day 7, respectively. The mRNA expression of CREG was also decreased, but the expression of cleaved caspase-3 at the mRNA and protein levels was increased obviously. Annexin V/PI FCM assay indicated that the apoptotic rate of R1-shCREG/EB was significantly higher those that in other 2 groups on day 7. CONCLUSION: The down-regulation of CREG inhibits ESCs differentiation and promotes cell apoptosis.     

高昼温对日光温室番茄叶片碳水化合物代谢的影响

以‘辽园多丽’番茄为试材,以昼温25℃为对照,对植株进行30℃和35℃高昼温处理,探讨在高昼温条件下,叶片碳水化合物含量、蔗糖代谢酶活性以及叶绿体超微结构的变化。结果表明：高昼温尤其是35℃昼温处理,番茄叶片叶绿体超微结构发生改变,短时间内叶绿体中淀粉粒数减少,持续15 d后则明显增多,与对照差异显著。高昼温处理的番茄叶片中果糖、葡萄糖和蔗糖含量均降低,昼间温度越高,各糖分含量与对照间差别越明显。在高昼温处理15 d时,叶片中酸性转化酶(AI)、中性转化酶活性(NI)降低,而蔗糖合成酶(SS)、蔗糖磷酸合成酶(SPS)活性升高,昼温越高,蔗糖代谢相关酶活性与对照的差别越大。高昼温改变了番茄叶片叶绿体的超微结构,并且对糖分的积累与代谢产生影响,最终对植株的生长发育及产量与品质的形成不利,昼温越高,所造成的不利影响越大。     

MdMYB73的分子克隆及其在苹果愈伤组织和拟南芥幼苗中的盐抗性功能鉴定

从‘嘎拉’苹果中克隆了一个MYB转录因子基因(序列号:MDP0000894463)。该基因包含长为729 bp完整的开放阅读框,编码243个氨基酸,预测其蛋白质分子量为26.34 kD,等电点为9.29。系统进化树分析表明,这一MYB转录因子与拟南芥AtMYB73同源序列相似性最高,因此将其命名为MdMYB73。功能域分析表明,MdMYB73蛋白含有保守的R2R3-typeMYB绑定域。荧光定量PCR分析表明,MdMYB73在苹果的各个组织均有表达,在叶片和花中表达相对较高;MdMYB73的表达明显受盐胁迫的诱导。将异位表达MdMYB73的拟南芥幼苗进行抗盐鉴定,结果表明MdMYB73负调控拟南芥盐胁迫抗性;同时,AtSOS1,AtSOS3和AtNHX1抗盐相关基因的表达水平显著降低,表明MdMYB73可能负调控SOS反应,影响拟南芥抵抗高盐胁迫过程。将MdMYB73基因遗传转化苹果愈伤组织,抗盐表型分析表明,MdMYB73过量表达也明显降低了转基因愈伤组织对盐胁迫的抗性。