基于cpDNA rps16序列分析兰考泡桐与白花泡桐和毛泡桐的遗传关系

【目的】通过测序法分析兰考泡桐与白花泡桐和毛泡桐在叶绿体rps16序列上的遗传差异,旨在分析三者之间在叶绿体基因上的变化特点和规律,探讨其种间的遗传关系。【方法】选取兰考泡桐、白花泡桐和毛泡桐各15个样本,对其提取的DNA用PCR扩增获得特异片段,并将其纯化与测序。利用软件Clustal X 2.0对所得序列进行排序;运行MEGA 4软件,进行多序列比对,分析其序列特征,并计算出K2P遗传距离。【结果】(1)对获得的rps16序列进行测定分析,得兰考泡桐序列长度分别为932 933 bp;白花泡桐序列长度为932 bp;毛泡桐序列长度分别为916918 bp。对所得rps16序列进行排序后的长度为938 bp,平均GC含量为34.31%。3个种所代表的个体之间共有10个变异位点,占整个序列长度的1.07%。其中有9个变异位点属于碱基插入或缺失类型,占变异位点总数的90%,占整个序列长度的0.96%。有1个变异位点属于碱基替换类型,占整个变异位点总数的10%,占整个序列长度的0.11%。(2)整个rps16片段的序列共有10个变异位点,其中兰考泡桐与白花泡桐在总的变异位点上,具有一致的碱基位点9个,占总变异的90%。而兰考泡桐与毛泡桐相比,没有相同的碱基。【结论】根据三种泡桐的rps16序列的序列特征和变异位点的分析,表明在叶绿体遗传方面,兰考泡桐具有与白花泡桐更多相似的遗传物质,其亲缘关系较近。综上所述,推测兰考泡桐与白花泡桐可能来自同一母系遗传。     

崖柏属植物的核型分析

[目的]为了从细胞学角度探讨崖柏属植物分类与进化问题,[方法]采用根尖压片法对崖柏属5种植物进行核型研究。[结果]表明:崖柏和日本香柏的核型公式为2n=2x=22=18m(2SAT)+4sm,朝鲜崖柏、北美乔柏和北美香柏的核型公式为2n=2x=22=20m(2SAT)+2sm,5种植物的核型均属于1A类型。[结论]该属在柏科中可能处于比较进步的进化地位;通过进化趋势图分析发现,崖柏在该属中分化最晚,而朝鲜崖柏则较原始。     

草木樨中华根瘤菌CHW10B溶磷特性及其对南方红豆杉的促生作用

[目的]探讨南方红豆杉根际微生物草木樨中华根瘤菌(Sinorhizobium meliloti)CHW10B的溶磷特性及促生作用。[方法]利用液体发酵试验比较不同时间、碳源、氮源、碳氮比、培养温度等条件下CHW10B菌株的溶磷量,采用温室盆栽法研究该菌株对南方红豆杉生长的影响,并分析该菌株产嗜铁素、精氨酸脱羧酶、ACC脱氨酶及有机酸等成分的能力。[结果]该菌株分别在培养时间4 d、磷酸钙5.00 g·L~(-1)、初始p H值7.0、装液量1/2、NaCl为0.0 g·L~(-1)、温度30℃、碳源为葡萄糖、氮源为硫酸铵、碳氮比100∶1时溶磷能力最强。将该菌株接种于1年生南方红豆杉实生苗360 d后,苗木的地径、苗高、生物量比对照分别增长了19.53%、20.14%、25.39%。该菌株能够产嗜铁素、精氨酸脱羧酶、ACC脱氨酶(0.922 U·mg~(-1))和IAA(8.908 mg·m L~(-1)),并可分泌大量有机酸—葡萄糖酸(5 704.92μg·L~(-1))。[结论]草木樨中华根瘤菌CHW10B溶磷能力强,对南方红豆杉促生作用显著,适用于多种不同酸碱性土壤,可高效应用于南方红豆杉的人工栽培,具有被开发为生物肥料的潜力。     

Potential of the entomopathogenic nematodes Steinernema feltiae , S. weiseri and Heterorhabditis bacteriophora for the biological control of the sugar beet weevil Bothynoderes punctiventris (Coleoptera: Curculionidae)

The sugar beet weevil, Bothynoderes punctiventris Germ. (Coleoptera: Curculionidae), is one of the most damaging pests of sugar beets in Turkey and has been traditionally controlled by application of large amounts of insecticides. The current study used laboratory microcosms to evaluate the possibility of using entomopathogenic nematodes as an alternative control method. The nematodes Steinernema feltiae (strain TUR-S3), Steinernema weiseri (BEY) and Heterorhabditis bacteriophora (TUR-H2) had previously been isolated from Turkey. Nematode-induced mortality generally increased as soil temperature increased from 15 to 25°C but decreased when larvae were located deeper in soil. Heterorhabditis bacteriophora caused the highest larval mortality at 25°C at all depths (5–20 cm). Steinernema feltiae and S. weiseri were more effective than H. bacteriophora at 15°C at all depths. Increasing the application rate of infective nematode dauer juveniles (DJs) affected the number of DJs that penetrated each insect larva and the number of DJs produced per insect. The highest production of DJs per larva occurred at application rates of 50 DJs/larva for S. feltiae and S. weiseri and 100 DJs/larva for H. bacteriophora. Reproduction decreased again at higher application rates. Heterorhabditis bacteriophora had the highest reproductive capability. The results indicate that S. feltiae and S. weiseri might be more effective against this pest early in the growing season when soil temperature is low and that H. bacteriophora might be more effective later in the season when temperature increases.     

A new norlignan from Taxodium ascendens

A new norlignan, (2R,3R,4S,5S)-2,4-bis(4-hydroxyphenyl)-3,5-dihydroxy-tetrahydropyran (1), together with 9 known compounds were isolated from the branches and leaves of Taxodium ascendens. Their structures were mainly determined on the basis of MS, IR, 1D and 2D NMR spectral evidences. Methanol extract showed inhibitory activity on carbonic anhydrase II with an IC₅₀ value of 4.27 µg/ml, acetone extract and methanol extract inhibited activity of cathepsin B with IC₅₀ values of 2.12 and 3.71 µg/ml, respectively.     

Effect of Lecanicillium muscarium on Eretmocerus sp. nr. furuhashii (Hymenoptera: Aphelinidae), a parasitoid of Bemisia tabaci (Hemiptera: Aleyrodidae)

Pathogenicity of Lecanicillium muscarium, against Eretmocerus sp. nr. furuhashii (Hymenoptera: Aphelinidae), was investigated under laboratory conditions to determine if the fungal infection of the whitefly host can effect the survival, longevity and fecundity of female parasitoid. The results indicated that the number of parasitized larvae surviving a L. muscarium treatment after 6 days of oviposition decreased with increasing concentrations of L. muscarium and in later stages of development (12 days post oviposition) were not affected by fungal application. There were no significant differences on adult parasitoid survivorship after 7 days among all treatments. Maximum survivorship (73.33%) was observed for control and it was minimum (60%) at 1 × 10⁸ conidia/ml. L. muscarium showed a non significant effect on longevity and next offsprings of female parasitoids. The percentage emergence of parasitoids from the whitefly nymphs produced by the females emerged from treated pupae was almost similar. Maximum emergence (69.77%) was observed at 1 × 10⁶ conidia/ml and it was lowest (61.02%) at conidial concentration of 1 × 10⁸ conidia/ml. Maximum longevity of adult Eretmocerus sp. emerging from whitefly nymphs when treated after 12 days of postoviposition was observed for 1 × 10⁵ conidia/ml having a mean value of 5 days whereas the lowest longevity was 4.9 days observed at 1 × 10⁸ conidia/ml. The results mentioned above indicate that the interaction among biocontrol agents is positive to a greater extent with minimum risk hazards.     

中间锦鸡儿CiDR1的克隆及干旱胁迫下的表达分析

[目的]研究并了解中间锦鸡儿CiDR1基因功能及其对干旱胁迫的响应,为抗性育种提供候选基因。[方法]通过RACE技术从中间锦鸡儿中克隆CiDR1基因的c DNA全长,利用生物信息学分析软件对其基因结构及功能进行分析和预测。再通过qRT-PCR技术对干旱胁迫后的幼苗中的CiDR1表达模式进行研究。[结果]从中间锦鸡儿中克隆到CiDR1基因的c DNA全长共计4 297 bp,Gen Bank登录号为KP277100。生物信息学分析表明,预测的CiDR1蛋白序列中含有1 243个氨基酸残基,具有抗病基因特征结构域TIR、NB-ARC、LRR等,其等电点为6.35,不稳定指数为42.91,不具备信号肽,为非分泌蛋白,定位于细胞质中。定量PCR检测发现,CiDR1基因在幼年期的茎中表达量较低,在成年期的叶片中表达量较高;在干旱胁迫处理后的幼苗中,CiDR1表达水平有明显下降,表明该基因的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关。[结论]中间锦鸡儿在干旱胁迫后其根、茎和叶中CiDR1的表达均明显下降,表明CiDR1的表达受干旱抑制,可能与中间锦鸡儿适应干旱相关,进一步研究发现CiDR1在根、茎、叶中的表达水平可能受发育阶段调控。     

巨桉EXP基因家族的生物信息学分析

[目的]本研究有助于了解EXP基因家族的基本特征,为深入研究其功能搭建平台。[方法]本研究对从巨桉(Eucalyptus grandis Hill)中筛选出35个EXP基因家族成员(Egr EXP1 Egr EXP35),利用生物信息学方法对其基因特征与表达模式进行综合分析。[结果]巨桉EXP基因分布在8条染色体之上,EXP蛋白均定位在细胞质膜上发挥作用,大多数的家族成员具有信号肽。巨桉EXP编码的蛋白质由α-螺旋、延伸链、无规卷曲、β-转角组成。进化分析结果表明,巨桉EXP蛋白与毛果杨(Populus trichocarpa) EXP蛋白的进化关系接近。35个巨桉EXP基因在巨桉未成熟木质部、成熟叶片、韧皮部、茎尖、木质部以及幼叶组织中表达模式存在显著差异。[结论]EXP基因家族各成员的表达模式不同,Egr EXP17、Egr EXP18可能在巨桉木材形成过程中起重要作用。     

Phytochemical and antifungal studies on Terminalia mollis and Terminalia brachystemma

Phytochemical investigation of the stem bark of Terminalia mollis afforded friedelin (1), catechin with epicatechin (2), gallocatechin with epigallocatechin (3) and 3-O-methylellagic acid 4'-O-α-rhamnopyranoside (4). Arjunolic acid with 2α, 3β, 23-trihydroxy-urs-12-en-28-oic acid (5), 2α-hydroxyursolic acid (6), gallic acid (7), chebulanin (8) and 2'-O-galloylvitexin (9) were isolated from the leaf. Chebulanin (8), betulinic acid (10), ursolic acid (11), catechin (12), isoorientin (13), orientin (14), isovitexin (15) and punicalagin (16) were isolated from Terminalia brachystemma leaf. The first full unambiguous NMR assignments for (4) and (8), and revised assignments for (9), are reported. Compound (16) showed good activity against three Candida species.     

苹果属无融合生殖SERK1基因的克隆与生物信息学分析

[目的]为探讨苹果属植物无融合生殖分子机制。[方法]以苹果属平邑甜茶及杂种后代33#为试材,以苹果基因组CDS序列设计引物,通过PCR扩增技术克隆出SERK同源基因的cDNA全长序列,命名为MhSERK1和MhdSERK1(GenBank登录号JQ231273和JQ231272),利用实时定量RTqPCR的方法检测了这两个基因在平邑甜茶和杂种后代各组织和器官中的表达模式。[结果]序列分析显示MhSERK1和MhdSERK1编码区序列全长为1 899 bp和1 881 bp,分别编码632和626个氨基酸,其氨基酸序列与其他植物的SERK1同源基因所编码的氨基酸同源性都在80%以上,特别是与葡萄科龙眼品种同源性最高,高达92.56%,与模式植物拟南芥、烟草等植物的SERK同源基因都具有很高的同源性。实时定量PCR结果表明,在平邑甜茶和杂种后代不同组织、花器官中SERK1基因的表达量存在差异,其中在子房中的表达量最高,在营养生长的组织中表达量很低,在平邑甜茶花蕾期的子房中表达量最高。[结论]推测该基因在平邑甜茶和杂种后代的生殖发育过程中可能发挥重要作用。     

黑木相思与尾叶桉苗期氮素传递的研究

以尾叶桉、黑木相思苗木混栽为研究对象,采用¹⁵N土壤和叶片标记法,在温室盆栽下研究2种植物间在根系完全隔离、部分隔离及无隔离处理下的生长变化及N素传递。结果表明:黑木相思的苗高、地径及生物量等生长指标随着根系隔离的减少而逐渐降低;相反,尾叶桉在根系无隔离处理下,其生长指标都要显著高于根系完全隔离处理,结果表明尾叶桉具有强大的养分吸收能力,混栽条件下会抑制黑木相思的生长。在¹⁵N土壤或叶片标记下,尾叶桉在根系部分隔离、无隔离下的总¹⁵N含量显著高于完全隔离处理(5.0和59.6倍);在土壤标记下,根系无隔离处理的黑木相思总¹⁵N含量显著低于完全隔离、部分隔离处理,而在叶片标记下的完全隔离黑木相思,其总¹⁵N含量显著低于其余2种处理。无论土壤或是叶片标记,黑木相思都能将N素传递至相邻尾叶桉,其N素传递率、传递量随着根系隔离的减少而提高。研究结果表明尾叶桉、黑木相思间能通过土壤渗透等方式传递N素,为桉树、相思的混交模式提供可靠的理论依据。     

基于MaxEnt模型的毛红椿适生区预测

[目的]了解环境因子对毛红椿适生区分布的影响,为其资源保护、引种及其人工林的发展提供参考。[方法]基于MaxEnt模型能利用现存不完整、小样本、离散型分布数据构建物种适生区预测模型,用受试者工作曲线线下面积(AUC)检验预测模型的精度,面积越大精度越高等优点,本研究应用毛红椿在云南的分布数据及1个地型因子和6个气候因子,来构建其适生区分布模型。[结果]毛红椿适生区分布MaxEnt模型平均训练AUC和平均测试AUC分别为0.891、0.885,说明对毛红椿适生区的预测是可靠的;降水量变异系数和最干季度降水量是决定毛红椿适生区分布的主要因子,年均气温变化范围、最冷季度平均气温、最湿季度降水量、最冷季度降水量是次要因子。在当代和未来(2050S、2070S)气候变暖条件下(RCP2.6情景),云南省和全国适生区面积计算结果直观、定量的反应了全球变暖对毛红椿适生区变迁的影响。[结论]预测云南省及全国的毛红椿适生区随全球变暖而小幅萎缩。     

淡紫拟青霉航天诱变菌株对南方根结线虫的致病力

[目的]为筛选对南方根结线虫具有高致病力的淡紫拟青霉航天诱变菌株,通过淡紫拟青霉山东寿光菌株搭载神舟八号获得10个淡紫拟青霉航天诱变菌株.[方法]以这10个航天诱变菌株对南方根结线虫卵进行寄生试实验,观察了其发酵液对南方根结线虫卵及二龄幼虫的作用,并对与毒力相关的胞外酶几丁质酶和蛋白酶活性进行了测定.在此基础上,通过盆钵试验方法, 检验菌株Sd-m-9、Sd-m-16和Sd-m-26对花椒南方根结线虫的防治效果.[结果]结果表明:10个诱变菌株对南方根结线虫卵的寄生率与原始菌株存在分化,其中有3个诱变菌株的寄生率增强,即Sd-m-9、Sd-m-16和Sd-m-26;这10个诱变菌株的几丁质酶和蛋白质酶的活性与对南方根结线虫卵的寄生率之间呈正相关.在盆钵试验中,菌株Sd-m-9、Sd-m-16和Sd-m-26对南方根结线虫的根结指数较原始菌株下降88.34%~89.70%.[结论]因此,航天诱变菌株Sd-m-9、Sd-m-16和Sd-m-26可用于对南方根结线虫的防治.     

松材线虫(Bursaphelenchus xylophilus)耐久型幼虫(LIV)携带的香茅醇假单胞杆菌(Pseudomonas citronellolis)分离与鉴定

[目的]目前有关松材线虫与伴生细菌的关系及伴生细菌的病原作用是松树枯萎病研究的重点。为了揭示松材线虫与伴生细菌之间存在的密切关系,作者对松材线虫L_IV幼虫携带的细菌进行了分离鉴定。[方法]根据培养性状和16S rDNA序列同源性以及系统发育学等方面进行分析鉴定。[结果]确定L_IV幼虫携带的是香茅醇假单胞杆菌（Pseudomonas citronellolis）,携带率为100%;每条L_IV幼虫携带量在1.4×10⁵~4.5×10⁵。L_IV幼虫生活在松褐天牛体内,是引起松材线虫病侵染流行的唯一虫态;新发现的香茅醇假单胞杆菌能分解纤维素及降解或合成萜烯和酚类化合物。[结论]L_IV幼虫携带香茅醇假单胞杆菌的发现,揭示了松树、松褐天牛、松材线虫、细菌同为一体的紧密关系,并为揭示松树枯萎病机制提供了一种新病原和重要的研究思路。     

毛竹和林下植被芒箕凋落物分解特征研究

[目的]探讨竹林与其林下植被凋落物叶之间相互影响的潜在机制,为合理经营管理毛竹林林下植被提供理论参考。[方法]采用原位分解袋法研究了四川长宁毛竹与林下植被芒箕凋落物叶分解和养分释放过程。[结果](1)芒箕凋落物叶初始C、N、P含量和羟基碳高于毛竹(P 0. 05),而C∶N、C∶P、烷基碳、氧烷基碳和芳香碳低于毛竹(P 0. 05)。(2)凋落物叶分解和养分释放速率芒箕整体高于毛竹,芒箕和毛竹分解常数(k)分别为0. 58±0. 03和0. 73±0. 02,C、N、P养分释放均表现为净释放。(3)凋落物叶混合对分解速率没有显著影响,但抑制了N、P元素整个分解周期和C元素中后期的释放。(4)凋落物叶分解过程中元素含量变化格局表现为C含量和C∶N比整体呈下降趋势,N含量和N∶P比有小幅上升,P含量有微弱的下降趋势,C∶P比呈波动性变化。(5)凋落物叶分解速率与土壤温度、初始凋落物叶N和P含量呈显著正相关(P 0. 01),与初始凋落物叶的C∶N和C∶P呈极显著负相关(P 0. 01),与土壤含水量相关不显著。[结论]单独分解过程中,毛竹凋落物叶分解速率低于林下植被芒箕,养分释放特征均表现为直接释放;混合分解过程中,毛竹和芒箕凋落物叶分解速率无显著混和效应,但养分释放的混合效应表现出一定负效应和不同阶段性。     

Two new furostanol saponins from Tribulus terrestris L.

Two new furostanol glycosides, named tribufurosides I (1) J (2), were isolated from the fruits of Tribulus terrestris L. by a combination of chemical and spectroscopic methods. Its structures were established as 26-O-β-d-glucopyranosyl-(25S)-5α-furost-12-one-2α,3β,22α,26-tetraol-3-O-β-d-glucopyranosyl (1 → 2)-β-d-glucopyranosyl (1 → 4)-β-d-galactopyranoside (1) and 26-O-β-d-glucopyranosyl-(25R)-5α-furost-20(22)-en-12-one-2α,3β,26-triol-3-O-β-d-glucopyranosyl (1 → 4)-β-d-galactopyranoside (2).     

铁皮石斛phytocyanin基因家族全基因组分析

[目的]探讨Phytocyanins(PCs)在铁皮石斛发育过程和逆境胁迫环境下的潜在功能。[方法]利用拟南芥和水稻phytocyanin基因家族蛋白序列(At PCs和Os PCs),采用本地化软件BLASTP对铁皮石斛全基因组数据库进行搜索,并采用SMART和pfam数据库验证质体蓝素样结构域,获得铁皮石斛phytocyanin基因家族编码序列(Do PCs);利用Signal P4.1、Big-PI Plant Predictor、Net NGlyc1.0 Server等在线软件对Do PC家族序列的结构特点进行分析;利用软件Clustal W和MEGA对Do PC基因进行氨基酸序列的比对和系统进化分析;利用转录组数据绘制heatmap图对Do PCs基因在与美孢胶膜菌共生萌发的种子中的表达情况进行分析。[结果]显示:铁皮石斛全基因组中共预测到包含PCLD结构域的38个Do PCs,可分属4个亚家族,2个Cys残基在Do PCs家族中高度保守;19条Do PCs蛋白序列含有4个完整的保守铜结合配体(His、Cys、His、Gln/Met),有32条Do PCs骨架中含有N-端信号肽,22条Do PCs含有糖基磷脂酰肌醇锚定位点,有27条Do PCs含N-糖基化位点;16个Do PCs基因在与美孢胶膜菌共生萌发的种子中检测到表达,其中,Do UCL2,4和Do ENODL14基因表达量最高,7个基因在共生萌发的种子中明显上调表达,只有Do ENODL6基因明显下调表达。[结论]在全基因组范围内分析铁皮石斛Do PC家族蛋白结构特点,可以为探究兰科植物与微生物共生的相互作用及进一步研究兰科菌根形成的分子机制提供数据基础。     

Two new flavonoids from Retama raetam

Two new flavones were isolated from the aerial parts of Retama raetam subsp. raetam. Their structures were established as luteolin 4′-O-neohesperidoside (1) and 5,4′-dihydroxy-(3″,4″-dihydro-3″,4″-dihydroxy)-2″,2″-dimethylpyrano-(5″,6″:7,8)-flavone (2) by means of spectroscopic methods. Also present was ephedroidin (4′,5,7-trihydroxy-8-(2-hydroxy-3-butenyl)-flavone).