A new diterpene from the leaves of Andrographis paniculata Nees

Phytochemical investigation of the ethanol extract of the leaves of Andrographis paniculata yielded one novel diterpene (13R, 14R) 3, 13, 14, 19-tetrahydroxy-ent-labda-8 (17), 11-dien-16, 15-olide 1 which has an uncommon cis-diol groups in the lactone moiety, and 3, 19-isopropylidene-14-deoxy-ent-labda-8 (17), 13-dien-16, 15-olide 2, probably an artifact diterpene, together with eight known diterpenoids 3–10. The structures of these compounds were determined on the basis of spectral methods. The structure and stereochemistry of 1 was confirmed by X-ray crystallographic analyses.     

银杏过氧化氢酶基因CAT1的克隆及表达分析

利用RACE技术从银杏中克隆到过氧化氢酶基因(GbCAT1)的cDNA全长。进化树分析结果表明:GbCAT1和其他物种的CAT源自于相同的祖先。Southern杂交显示:GbCAT1属于1个小的多基因家族。实时定量RT-PCR分析表明:GbCAT1在银杏的根、茎、叶和果中都有表达,在叶中的相对表达量最高,其次为果、茎和根。GbCAT1的转录受到ABA、渗透压、紫外、低温和高温胁迫的诱导。水杨酸处理下,GbCAT1相对表达量迅速降低。CAT1基因在逆境条件下的相对表达变化与环境胁迫有关。     

荔波连蕊茶肌动蛋白基因全长cDNA的克隆及其表达分析

根据植物肌动蛋白(Actin)基因编码区的保守序列设计引物,以山茶属植物荔波连蕊茶幼嫩茎段为材料,提取总RNA,进行RT-PCR。采用RACE技术扩增获得1 631 bp的Actin基因全长cDNA序列,命名为ClActin1(GenBank登录号KF366912)。序列分析表明,ClActin1开放阅读框(ORF)为1 134 bp,编码377个氨基酸,5’非编码区90 bp,3’非编码区407 bp。预测的ClActin1蛋白分子量为41.69 kD,理论等电点为5.31,具有Actin超基因家族的保守结构域和肌动蛋白家族特有的特征信号序列。ClActin1与GenBank中收录的其它植物肌动蛋白核苷酸序列的相似性在82%以上,氨基酸序列的相似性在97%以上。与其它植物肌动蛋白比较并构建系统进化树,结果显示山茶肌动蛋白与茶树和杨树的肌动蛋白的亲缘关系最为密切。实时荧光定量PCR结果显示,该基因在荔波边蕊茶不同组织器官及不同发育时期的表达量稳定,表明ClActin1基因可作为内参基因。     

松墨天牛在云南松树干的垂直分布研究

松墨天牛(Monochamus alternatus Hope) (又名松褐天牛、松天牛)属鞘翅目,天牛科,墨天牛属,是松属树种如马尾松(Pinus massoniana Lamb.)、黑松(P.thunbergii Parl.)、云南松(Pinus yunnanensis Franch)和思茅松(P. kesiya var. langbianensis(A.chev.)Gaussen)等的重要蛀干害虫,主要危害生长衰弱的树木或新伐倒木,造成松树枯死,同时还传播松材线虫(Bursaphelenchus xylopilus (Steiner & Buhrer) Nickle)萎蔫病[1-2].松墨天牛在云南主要危害云南松,目前分布范围已经扩展到43县(市),并在部分地方成灾,加大了松材线虫病传入的可能性[3].松墨天牛在滇中地区1年1代,雌成虫在树干上咬出长椭圆刻槽,将卵产在刻槽中部的表皮层下,幼虫生长发育至一定阶段蛀入木质部, 在边材表面留下椭圆形的蛀入孔;蛀入木质部的幼虫在蛀道内取食,老熟幼虫在蛀道末端构筑蛹室化蛹,成虫从蛹室向外咬出近圆形的羽化孔爬出[4-5].松墨天牛是典型的钻蛀性害虫,调查和防治非常困难.前人曾经对松墨天牛在马尾松和思茅松上的分布规律进行了研究[6-8],但这些研究是某一时间段天牛的危害痕迹或幼虫在树干上的分布,对于天牛产卵刻槽、蛀入孔和羽化孔的数量关系,目前还没有相关报道.本文对松墨天牛的产卵刻槽、蛀入孔和羽化孔在云南松树干的垂直分布规律及其数量关系进行了调查分析,旨在为松墨天牛的林间调查和防治提供参考.     

双斑长跗萤叶甲对几种植物挥发物的触角电位和行为反应

采用触角电位(EAG)、"Y"型嗅觉仪和风洞的方法研究双斑长跗萤叶甲成虫对14种植物挥发物的电生理和行为反应,筛选出对双斑长跗萤叶甲有效的植物挥发物,为该害虫的种群监测和防控提供新技术。研究结果表明:稀释102、103倍的薄荷精油和1 mol.L-1的香叶醇能引发双斑长跗萤叶甲强烈的EAG反应(P<0.01),且嗅觉测试表明,薄荷精油(稀释102倍)对双斑长跗萤叶甲的最大驱避率为(74±5.47)%,香叶醇(1 mol.L-1)对双斑长跗萤叶甲的最大引诱率为(70±10.00)%。风洞行为趋性研究表明:稀释102倍的薄荷精油对双斑长跗萤叶甲的最大驱避率为(72±8.37)%,而1 mol.L-1的香叶醇对双斑长跗萤叶甲的最佳引诱率为(66±8.94)%。     

杜仲1-脱氧-D-木酮糖-5-磷酸还原异构酶基因cDNA全长克隆与序列分析

以杜仲叶片cDNA为模板,采用反转录RCR及RACE技术分离出DXR基因cDNA全长。序列分析结果表明该基因序列全长1 814 bp,共编码478个氨基酸,推导的蛋白质分子量为51.71 kD,理论等电点5.79,命名为EuDXR。推导的EuDXR蛋白质序列具有植物DXR酶的5个典型基序,并预测出26个潜在的功能位点。系统进化树分析表明EuDXR蛋白与玉米、水稻亲缘关系最近,其次为橡胶、拟南芥、烟草。     

宽叶杜香叶柄再生体系建立及种质离体保存研究

The tender leafstalks of Ledum palustre var.dilatatum were used as explants for the experiment.Uniform design for the most suitable media for shoots regeneration immediately at base of tender leafstalks,rooting and germplasm conservation in vitro was screened.The results showed that N6+ZT 2.65 mg·L-1+IAA 0.05 mg·L-1 was fits for shoots regeneration,the frequency of shoots induction was higher than 92.5%;MS(modified)+IAA 0.1 mg·L-1+Kt 0.75 mg·L-1 for rooting,the rate of rooting was 98%;N-68+B9 2.5 mg·L-1+ Ph...     

Photosynthetic characteristics of Fagus sylvatica and Quercus robur established for stand conversion from Picea abies

Efforts in Europe to convert Norway spruce (Picea abies) plantations to broadleaf or mixed broadleaf-conifer forests could be bolstered by an increased understanding of how artificial regeneration acclimates and functions under a range of Norway spruce stand conditions. We studied foliage characteristics and leaf-level photosynthesis on 7-year-old European beech (Fagus sylvatica) and pedunculate oak (Quercus robur) regeneration established in open patches and shelterwoods of a partially harvested Norway spruce plantation in southwestern Sweden. Both species exhibited morphological plasticity at the leaf level by developing leaf blades in patches with an average mass per unit area (LMA) 54% greater than of those in shelterwoods, and at the plant level by maintaining a leaf area ratio (LAR) in shelterwoods that was 78% greater than in patches. However, we observed interspecific differences in photosynthetic capacity relative to spruce canopy openness. Photosynthetic capacity (A₁₆₀₀, net photosynthesis at a photosynthetic photon flux density of 1600 μmol photons m⁻² s⁻¹) of beech in respect to the canopy gradient was best related to leaf mass, and declined substantially with increasing canopy openness primarily because leaf nitrogen (N) in this species decreased about 0.9 mg g⁻¹ with each 10% rise in canopy openness. In contrast, A₁₆₀₀ of oak showed a weak response to mass-based N, and furthermore the percentage of N remained constant in oak leaf tissues across the canopy gradient. Therefore, oak photosynthetic capacity along the canopy gradient was best related to leaf area, and increased as the spruce canopy thinned primarily because LMA rose 8.6 g m⁻² for each 10% increase in canopy openness. These findings support the premise that spruce stand structure regulates photosynthetic capacity of beech through processes that determine N status of this species; leaf N (mass basis) was greatest under relatively closed spruce canopies where leaves apparently acclimate by enhancing light harvesting mechanisms. Spruce stand structure regulates photosynthetic capacity of oak through processes that control LMA; LMA was greatest under open spruce canopies of high light availability where leaves apparently acclimate by enhancing CO₂ fixation mechanisms.     

日本落叶松体细胞胚胎发生相关基因LaSERK1的克隆与表达分析

本研究克隆了日本落叶松体细胞胚胎发生相关受体类蛋白激酶基因 LaSERK1,并检测了在不同培养条件下LaSERK1的表达情况,发现LaSERK1与已发现的其他物种SERK1 基因有高度相似性,qRT-PCR结果显示： LaSERK1在体胚发生早期高表达。结果表明：LaSERK1可能在落叶松体胚发育早期起重要作用,LaSERK1 也可能成为早期胚性细胞的标记基因。     

猴头菌锰过氧化物酶1基因在构巢曲霉的异源转化与表达

为提高猴头菌菌株CB1锰过氧化物酶(MnP)基因的表达产量,采用PEG/CaCl2介导的原生质体转化方法,将携带有He-mnp1的重组质粒pLB01/He-mnp1转入到构巢曲霉尿嘧啶尿苷营养缺陷菌株TN02A7的原生质体中,获得了转化子菌株TN02A7-He-mnp1,并在乙醇脱氢酶启动子alcA(p)控制下实现了异源表达。将TN02A7-He-mnp1、TN02A7、构巢曲霉野生型菌株WJA01、猴头菌菌株CB1在相同的木质素环境下进行培养并检测MnP酶活性,结果表明:转化子菌株TN02A7-He-mnp1在0.05 g.L-1血红素的情况下、诱导96 h后酶活性最高为38.31 U.L-1,比不添加血红素的酶活力高8.64倍,但比猴头菌菌株CB1的酶活力低,而TN02A7与WJA01始终无MnP酶活性,说明基因He-mnp1已经成功地被转化到TN02A7-He-mnp1中,并在木质素环境下得到表达,血红素是重组MnP基因异源表达的限制性因素之一。本文为生产MnP和提高MnP产量提供了新的途径。     

杨树生物钟节律基因PtCCA1的克隆及表达模式研究

生物钟节律基因CCA1在植物的光周期反应中起着重要的调控作用.利用RT-PCR方法克隆获得了一条包含1902 bp开放阅读框的杨树CCA1基因cDNA序列,其编码633个氨基酸残基,蛋白的分子量约为68.90 kDa,等电点为6.33.分析显示含有1个Myb-like DNA结合域及2个蛋白跨膜结合域.实时荧光定量PCR分析发现,PtCCA1基因主要在杨树叶组织中表达;随着光照时间的变化,该基因在杨树中的表达量呈现白昼不断降低而夜晚逐渐升高的昼夜变化趋势.研究结果为进一步探索PtCCA1基因在调控杨树光周期响应途径中的功能以及杨树光周期敏感机制提供了科学基础.     

neo-Clerodane diterpenes from Ajuga decumbens and their inhibitory activities on LPS-induced NO production

A phytochemical investigation of the whole plants of Ajuga decumbens led to the isolation of three new (1, 2a, and 2b) and three known (3a−3c) neo-clerodane diterpenes. Their structures were elucidated by spectroscopic data analysis (IR, ESIMS, HR-ESIMS, 1D and 2D NMR), and the structure of 1 was confirmed by X-ray crystallography. The inhibitory activities on LPS-induced NO production of the six diterpenes were evaluated and compounds 2a, 2b, and 3a showed inhibitory effects.     

Sesquiterpenes from the aerial part of Chloranthus japonicus and their cytotoxicities

Four new sesquiterpenes, chlorajapolides F–I (1–4), along with nine known terpenoids (5–13) were isolated from the aerial part of Chloranthus japonicus. Their structures were elucidated on the basis of spectroscopic analysis, and a lindenane sesquiterpene, named 9-hydroxy-heterogorgiolide, previously isolated from the C. japonicus, was revised as its 8-epimer (1a). Moreover, methanol extract (ME), EtOAc fraction (EF), water fraction (WF), and all isolates (1a, 1–13) were evaluated for their cytotoxicities using two human cancer cell lines.     

毛竹PeAP2基因及其启动子的克隆与表达初步分析

APETALA2(AP2)基因在植物生长发育过程中发挥着重要作用.利用RT-PCR和RACE方法,从毛竹中克隆到1个AP2同源基因的全长cDNA序列,命名为PeAP2.序列分析表明:PeAP2基因全长1 750 bp,其中,5′端非编码区106bp,3′端非编码区174 bp,开放阅读框1 470 bp,编码1个489 aa的蛋白,该蛋白含有2个AP2结构域,属于AP2/EREBP家族的AP2亚家族.PeAP2蛋白与来自其它单子叶植物的AP2蛋白均有着较高同源性,其中,与二穗短柄草的AP2蛋白同源性最高,达74.85％.实时定量PCR分析显示:PeAP2基因在毛竹的根、茎、叶、鞘和节5种器官中均有表达,其中,叶片中的表达丰度最高,鞘中次之,而在根、茎、节中的表达丰度接近,均较低.利用hiTAIL-PCR方法克隆获得了PeAP2上游启动子区序列1 359 bp,分析显示其含有光、激素等多种信号应答相关的作用元件.     

马尾松钙调素基因的克隆及响应松材线虫侵染的表达特征

松材线虫病是由松材线虫(Bursaphelenchus xylophilus)侵染引起的严重病害,引起重大的经济和生态损失。钙调素(CaM)是真核生物中Ca2+主要传导蛋白,本研究采用RT-PCR方法,首次从马尾松中克隆获得CaM,命名为pmCaM,并检测了其响应松材线虫侵染的表达特征。序列分析表明:pmCaM完整的开放阅读框核苷酸序列为450bp,编码149个氨基酸的蛋白质;该蛋白含有4个EF-hand结构域,具有钙调素的典型特征,与其它植物的CaM蛋白均有较高同源性。实时荧光定量PCR分析显示:接种松材线虫后30 180 min时间段,马尾松苗根、茎、叶器官中的pmCaM均下调表达,但不同器官间显著下调表达的时间点存在差异;同时,不同器官pmCaM表达量变化均存在特异的随时间发展的波动特征,其中发现,根茎pmCaM在松材线虫接种45 min时均处于表达量高峰。本研究结果显示pmCaM表达响应了松材线虫侵染,揭示pmCaM可能参与了调控松材线虫-马尾松互作早期的钙信号响应。     

三年桐、千年桐感染枯萎病病原菌后的生理反应

枯萎病是油桐毁灭性病害。中国广泛种植的油桐有三年桐和千年桐,三年桐易感枯萎病,千年桐抗枯萎病。为探讨三年桐与千年桐在枯萎病应答过程中的差异,在对油桐枯萎病灾区调研的基础上,进行枯萎病病原菌的分离、鉴定,并进一步探讨三年桐、千年桐接种枯萎病病原菌后超氧化物歧化酶(SOD)、过氧化物酶(POD)、过氧化氢酶(CAT)活性和丙二醛(MDA)含量的变化。结果表明:油桐枯萎病病原菌为尖孢镰刀菌,接种试验显示千年桐在接种尖孢镰刀菌后SOD、POD、CAT较三年桐均呈现较高活性,在接种后SOD、POD活性先升高后降低,CAT活性升高并维持高活性,MDA含量变化不明显;三年桐SOD活性和MDA含量均先升高后降低,POD活性先降低再升高,CAT活性先降低后升高又降低。千年桐作为抗病种,其抗病性可能与其本身具有较高的SOD、POD、CAT活性有关,并且和病原菌感染后酶活上升有关;三年桐为易感病种,接种后SOD、POD、CAT活性有变化,但仍不能抵御病原菌的危害。     

30个桉树无性系人工林碳储量分析

通过对国营雷州林业局30个5年生桉树无性系人工林的调查、试验,旨在阐明不同桉树无性系人工林碳储量的变化规律及营建桉树碳汇林的合理措施.结果表明:30个桉树无性系人工林生态系统平均碳储量为148.743 t·hm-2,高于之前学者研究的桉树人工林碳储量,其中,乔木层和土壤层分别占34.39％、61.88％;乔木层平均碳储量达51.948 t·hm-2,不同无性系间差异极显著(p＜0.01),其中,23(101-1)、25(179-1)、4(BU1)、26(184-1)号无性系表现最优;土壤层的平均碳储量为92.033 t·hm-2,不同无性系土壤层碳储量差异不明显;灌木层、草本层、凋落物层碳储量分别是2.430、0.731、1.592 t·hm-2,占比例较小.营建桉树碳汇林关键在于无性系的正确选择.     

A new dihydrodibenzodioxinone from Hypericum x ‘Hidcote’

One new compound, 4-hydroxy-4a,7-dimethoxy-4,4a-dihydrodibenzo-p-dioxin-2(3H)-one (1), was isolated from the aerial parts of the hybrid Hypericum x ‘Hidcote’, together with 8 known compounds: caryophyllene-4,5-epoxide, quercetin, quercitrin, quercetin-3-O-β-D-galactopyranoside, epicatechin, betulinic acid methyl ester, β-sitosterol and β-sitosterol glucoside. The structure of the new compound, as well as its absolute configuration, was established by means of spectroscopic data analyses, including 2D NMR spectroscopy and X-ray structural analysis.     

Sesquiterpenoids from Carpesium divaricatum and their cytotoxic activity

In the course of searching for cytotoxic terpenoids from medicinal plants in China, two new eudesmane sesquiterpenoids, 5α-hydroxy-13-methoxy-7αH,11αH-eudesm-4(15)-en-12,8β-lactone (1) and 1β-hydroxy-7αH,11αH-eudesm-4(15)-en-12,8β-lactone (2), along with fourteen known sesquiterpenoids were isolated from the whole plant of Carpesium divaricatum. The structures of new compounds were determined using spectroscopic methods, including IR, HRESIMS, and 1D and 2D NMR spectroscopy. The cytotoxicity of selected sesquiterpene lactones against human oral epidermoid carcinoma (KB), human breast cancer (MCF-7) and human hepatoma (HepG-2) cells was also evaluated by MTT method.