慈竹木质素合成酶基因4CL RNAi载体构建与烟草转化

根据慈竹4-香豆酸辅酶A连接酶(Na4CL)全长cDNA序列,通过RT-PCR从慈竹幼笋cDNA中扩增约600 bp的保守片段。DNAMAN多重比对分析表明,该目标片段与3条烟草4CL基因同源性达84.08%。将目标片段插入到pSK-int中间表达载体中,获得pSK-4CL-RNAi中间表达载体,然后将其转入植物双元表达载体pBI121中,构建该基因的shRNA表达载体,并利用农杆菌介导法转化烟草(K326)。对获得的抗性再生植株的PCR检测分析初步表明该基因已插入烟草基因组中。     

麻疯树pepc基因正义、反义植物表达载体构建与功能初步分析

根据麻疯树pepc基因和植物表达载体的酶切位点特征,分别将pepc基因全长序列3 000 bp正向插入pCAMBIA2300,构建了正义表达载体pCAMBIA-Jcpepc,基因片段597 bp反向插入pBI121构建了反义表达载体pBIJcpepc.通过农杆菌介导,采用叶盘法转化烟草,通过对转基因植株的PCR和PCR...     

油桐桐酸合成酶基因克隆和植物表达载体构建

α-桐酸合成酶是控制亚油酸向α-桐酸(18:3Δ9cis,11trans,13trans)转化的关键酶。本研究以发育中的油桐种子为材料,利用改良TRIzoL法提取总RNA,并根据GenBank中已经登录的α-桐酸合成酶基因FADX的序列,设计特异性引物,采用RT-PCR方法,克隆得到FADX基因全长cDNA,长度为1221bp。通过生物信息学分析结果表明,该序列5’端和3’端非编码区序列长度分别为13bp和47bp,含有一个开放阅读框(14～1174bp),编码386个氨基酸,含有典型的脂肪酸脱氢酶结构域。将所得基因经BamHI和SacI酶切后插入表达载体质粒pBI121,构建反义表达载体pBI121fadx。     

桑树景天庚酮糖-1,7-二磷酸酶基因的克隆、原核表达与植物表达载体的构建

景天庚酮糖-1,7-二磷酸酶(SBPase)是卡尔文循环过程中的关键酶.利用RACE技术得到景天庚酮糖-1,7-二磷酸酶基因全长cDNA,命名为MSBPase(GenBank登录号;DQ995346).MSBPase全长为1 527 bp,该序列含有1个1 179 bp的完整开放读码框,编码393个氨基酸,蛋白质理论分子质量约为42.6 ku,等电点为5.85,其氨基酸序列与其他植物中已分离的SBPase有很高的同源性.对MSBPase编码的蛋白质(命名为MSBPase)进行结构预测分析表明,该蛋白富含无规卷曲(coil),高达64.29%,其次是α-螺旋(helix),为22.19%,而β-折叠(strand)只有13.52%.将MSBPase编码区插入原核表达载体pET30a( ),并转化到大肠杆菌菌株 BL21中,经过IPTG诱导,MSBPase融合蛋白在BL21菌株中成功表达.将得到的MSBPase编码区插入植物表达载体pBI121中,构建了MSBPase植物表达载体pBI121.SBP.     

国槐苯丙氨酸解氨酶基因的克隆、反义表达载体构建及遗传转化

根据其它植物PAL基因的保守区域,设计一对兼并引物,采用RT-PCR方法从国槐中克隆了一个长866 bp的PAL基因片段,命名为SjPAL.序列分析发现SjPAL多肽与其它植物的PAL氨基酸序列高度同源(87%以上),且包含与水稻和拟南芥的PAL类似的活性位点.系统进化树分析表明SjPAL与豆科植物的PAL亲缘关系较近.RT-PCR结果显示,SjPAL在根和茎的表达量约为叶中的3倍.利用反义RNA技术将SjPAL基因克隆至植物表达载体pBI121,构建了SjPAL反义RNA植物表达载体pBI121-PAL,通过根癌农杆菌EHA105将其导入拟南芥基因组,对获得的抗性植株进行PCR鉴定、Northern杂交分析、PAL活性分析以及总多酚含量和类黄酮含量分析.结果表明,该反义RNA已整合到拟南芥基因组中,转基因拟南芥的PAL基因表达量、单位材料PAL活性、总多酚含量和类黄酮含量均显著低于野生型对照.本研究为下一步利用该基因反义表达载体转化国槐,通过调节酚类物质含量提高其在再生体系中的生根能力提供了试验依据.     

美洲黑杨木质素合成关键基因的克隆及反义表达载体的构建

以美洲黑杨Populus deltoides新萌叶片为材料,通过自行设计引物,用RT—PCR的方法克隆了美洲黑杨木质素合成关键基因4CL和CAD基因部分序列。测序结果表明:这两个基因片段长度分别为859 bp和493 bp。此外,人工合成了4CL基因木质部特异表达启动子4CLp,长度为1 180 bp。分别用限制性内切酶Hind III/BamH I、BamHI/Sma I和Sma I/Sac I对4CLp、4CL和CAD基因序列进行双酶切,同时用Hind III/Sac I对pBI121表达载体进行双酶切,回收了目的片段。酶切回收后用T4DNA连接酶克隆到植物表达载体pBI121中,并转化至大肠杆菌感受态细胞DH5α。经质粒PCR和双酶切分析,确定获得了pBI—4CLp—a4CL—aCAD反义植物表达载体。     

马尾松纤维素合成酶基因PmCesA2的克隆及其植物表达载体的构建

为了探究马尾松木材形成的分子机制,以马尾松嫩枝的总RNA反转录得到的cDNA为模板,利用反转录PCR和RACE技术克隆得到马尾松CesA2基因的全长cDNA序列(命名为PmCesA2),序列长度为3 500 bp,包含一个长为3 174 bp的开放阅读框(open reading frame,ORF),编码1 057个氨基酸.序列分析表明:PmCesA2序列与已报道的火炬松CesA2基因(AY789651.1)的相似性达98％.将全长基因克隆到载体pBI121质粒的SnaBI,Sacl双酶切位点之间,构建正义表达栽体pBI121-PmCesA2.     

绿竹锌指蛋白基因BoBZF克隆及功能初步分析

锌指蛋白在原核生物与真核生物基因转录调控中发挥着重要作用.采用RT-PCR和RACE技术,从绿竹叶片中分离到1个锌指蛋白基因,命名为BoBZF(GenBank登记号:EU606025).BoBZF cDNA全长1 076 bp,编码256个氨基酸.蛋白结构分析表明,其编码的蛋白具有2个B-Box结构域,属于B-Box型锌指蛋白.组织特异性表达显示BoBZF在叶片、叶鞘、幼茎和根中均有表达,其中在叶片的表达丰度较高.将BoBZF置于CaMV 35S启动子控制下,构建到载体pBI121的多克隆位点,并转化拟南芥.RT-PCR分析表明,BoBZF已在拟南芥中表达.转基因植株耐旱性明显提高,意味着BoBZF与竹子的耐旱能力有关.     

RNA干涉培育低木质素杨树

采用改良的CTAB法提取南林95杨的基因组DNA,经PCR扩增得到肉桂酰辅酶A还原酶CCR基因的第4个外显子部分序列,通过中间载体pUCCRNAi,构建含正反向干涉片段的pBI121表达载体,导入农杆菌LBA4404。利用叶盘法侵染南林95杨,获得3株转基因植株,经分子鉴定证实干涉片段已导入南林95杨。测定Klason木质素及综纤维素含量的结果显示:转基因植株Klason木质素含量与对照相比平均降低了9.86%,综纤维素含量与对照相比平均增加了3.17%,纤维长宽比明显增加,均表明转基因植株更有利于造纸。     

杨树维管组织特异启动子的克隆与启动活性分析

启动子在基因表达调控中起关键性作用,它在很大程度上决定所控基因表达的时间、空间和强度。依据拟南芥ATH1芯片分析杨树维管形成时期特异表达基因的结果,选取了差异表达基因NST3,通过BLAST比对在杨树EST数据库(PopulusDB)中找到同源性较高的基因NAC068。以毛白杨为材料,在其基因组中克隆得到该基因5'侧翼区901 bp长的片段,命名为pProNAC068,将该片段置换pBI121载体中的CaMV35S启动子,并在84K杨中检测报告基因GUS的表达情况。经过GUS活性检测分析发现:该启动子可以控制外源基因在次生维管组织中特异表达,从而为基因工程中有目的的控制外源基因在维管组织中的表达奠定基础。     

C2H2-reduction byErythrina poeppigiana in a Costa Rican coffee plantation

Acetylene reduction activity by root nodules of the legumeErythrina poeppigiana, growing as shade tree in a Costa Rican coffee plantation, was estimated. The mean activity found was 15.7 nmole C₂H₄ · mg (dry weight)^–1 · h^–1. Root nodules collected at different distances from theErythrina stem showed the same activity per dry weight unit. However, as the biomass of the nodules was highest near the stem, was the acetylene reduction activity (expressed per soil volume) maximal near theErythrina stem and declined with distance.     

S-Lokalisation in SO2-begasten Fichtennadeln

Localisation of sulphur in spruce needles treated with SO₂. After exposure to SO₂ the tips of spruce needles arc richer in sulphur compounds than the bases. The accumulation of sulphur compounds results mainly in an increased content of inorganic sulphur. The analytical results explain the local occurrence of injuries at the needle tips.     

Stem radial CO_2 conductance affects stem respiratory CO_2 fluxes in ash and birch trees

The CO_2 released from respiring cells in woody tissues of trees can contribute to one of three fluxes:efflux to the atmosphere(E_A),internal xylem sap transport flux(F_T),and storage flux(DS).Adding those fluxes together provides an estimate of actual stem respiration(R_S).We know that the relative proportion of CO_2 in those fluxes varies greatly among tree species,but we do not yet have a clear understanding of the causes for this variation.One possible explanation is that species differ in stem radial CO_2 conductance(g_c).A high g_c would favor the E_A pathway and a low g_cwould favor the F_Tpathway.However,g_chas only been measured once in situ and only in a single tree species.We measured g_cusing two methods in stems of Fraxinus mandshurica Rupr.(ash)and Betula platyphylla Suk.(birch)trees in situ,along with R_S,E_A,F_T and DS.Stem radial CO_2 conductance was substantially greater in ash trees than in birch trees.Corresponding to that finding,in ash trees over 24 h,E_Aconstituted the entire flux of respired CO_2 ,and F_Twas negative,indicating that additional CO_2 ,probably transported from the root system via the xylem,was also diffusing into the atmosphere.In ash trees,F_T was negative over the entire 24 h,and this study represents the first time that has been reported.The addition of xylem-transported CO_2 to E_A caused E_Ato be 9% higher than the actual R_Sover the diel measurement period.Birch trees,which had lower g_c,also had a more commonly seen pattern,with E_A accounting for about 80% of the CO_2 released from local cell respiration and F_T accounting for the remainder.The inorganic carbon concentration in xylem sap was also lower in ash trees than in birch trees:2.7 versus 5.3 mmol L^-1,respectively.Our results indicate that stem CO_2 conductance could be a very useful measurement to help explain differences among species in the proportion of respired CO_2 that remains in the xylem or diffuses into the atmosphere.     

应用简化生物圈模式（SiB2）模拟通榆地气通量研究

主要通过对比观测与SiB2模拟的吉林通榆农田和草地样地的感热通量、潜热通量、CO2通量和气温来检测独立版简化生物圈模式的数值模拟能力。在模拟中,农田和草地代表半干旱区2种典型的地表覆盖类型,它们的全年冠层高度和叶面积指数都处于变化当中,在非生长季地表裸露,而在生长季,冠层高度农田可达2m,草地可达0.8m,对应的叶面积指数分别可达4.2和2.4。SiB2模型模拟用观测值为初始场,以半小时气象观测为驱动场。对2003年的模拟和观测的比对研究表明:在生长季模式具有很好的模拟小时时间尺度的潜热、感热和CO2通量及地表气温的能力;(2)模式模拟土壤热通量可能偏大;(3)模拟的逐日潜热、感热和CO2通量及地表气温可用合理地反映全年的季节变化。图11表3参23。     

2,6-二甲基-7-辛烯-2-烷基(或羟烷基)醚的合成研究

以氢型丝光沸石为催化剂，二氢月桂烯与单醇或二醇加成反应生成2，6-二甲基-7-辛烯-2-烷基或羟烷基醚。实验结果发现，各种醇与烯生成醚化合物的趋势如下：直链伯醇易于支链伯醇；伯醇易于仲醇；低碳直链伯醇易于高碳直链伯醇，同时还发现，1，3-丁二醇、1，6-己二醇与烯的反应，只生成单醚。文中还考察了反应温度和时间对反应的影响。     

橡胶优良品种海垦2在云南的适应性

橡胶树优良品种海垦 2在云南省热带作物科学研究所、勐捧农场、孟定农场的适应性试验研究表明 ,1～ 11割年平均单株年产干胶 5.2 7kg ,是对照RRIM6 0 0的 12 2 .8% ;单位面积产量为 2 0 96kg/hm2 ,是对照RRIM6 0 0的138.5%。海垦 2干胶含量高 ,产胶潜力大 ,可以在云南省一类型植胶区大规模种植     

福建省西番莲科2种新记录归化植物

报道了西番莲科2种福建省新记录归化植物,分别是细柱西番莲(Passiflora gracilis Jacq.ex Link)、三角西番莲(Passiflora suberosa L.).     

Functional involvement of Ca(2+) and Ca(2+)-activated K(+) channels in anethol-induced changes in Ca(2+) dependent excitability of F1 neurons in Helix aspersa

The effects of anethol, the major component of anise oil, on the Ca²⁺-dependent excitability and afterhyperpolarization (AHP) in snail neurons were examined using intracellular recording. Anethol (0.5%) significantly broadened the spike, reduced the firing frequency and enhanced the AHP amplitude. In contrast, anethol (2%) significantly increased the firing frequency and decreased the AHP. Blockade of Ca²⁺ channels after anethol application depolarized the membrane potential and significantly reduced the firing rate. Furthermore, in the presence of anethol (0.5%) a significant decrease in the AHP was observed by Ca²⁺ channels blockage. Here, anethol-induced functional modification of Ca²⁺ and Ca²⁺-activated K⁺ channels is suggested.     

Aluminum-induced decrease in CO2 assimilation in citrus seedlings is unaccompanied by decreased activities of key enzymes involved in CO2 assimilation

'Cleopatra' tangerine (Citrus reshni Hort. ex Tanaka) seedlings were irrigated daily for 8 weeks with 1/4 strength Hoagland's nutrient solution containing 0 (control) or 2 mM aluminum (Al). Leaves from Al-treated plants had decreased CO2 assimilation and stomatal conductance, but increased intercellular CO2 concentrations compared with control leaves. On a leaf area basis, 2 mM Al increased activities of key enzymes in the Calvin cycle, including ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), NADP-glyceraldehyde-3-phosphate dehydrogenase (GAPDH), phosphoribulokinase (PRK), stromal fructose-1,6-bisphosphatase (FBPase), and a key enzyme in starch synthesis, ADP-glucose pyrophosphorylase (AGPase), compared with control leaves. Aluminum had no effect on cytosolic FBPase activity, but it decreased sucrose phosphate synthase (SPS) activity. Aluminum had no effect on area-based concentrations of carbohydrates, glucose-6-phosphate (G6P) and fructose 6-phosphate (F6P) or the G6P:F6P ratio, but it decreased the area-based concentration of 3-phosphoglycerate (PGA). Photochemical quenching coefficient (qP) and electron transport rate through PSII were greatly reduced by Al. Non-photochemical quenching coefficient (NPQ) was less affected by Al than qP and electron transport rate through PSII. We conclude that the reduced rate of CO2 assimilation in Al-treated leaves was probably caused by a combination of factors such as reduced electron transport rate through PSII, increased closure of PSII reaction centers and increased photorespiration.