地黄源库关系的变化及其与连作障碍的关系

通过正茬剪叶模拟连作地黄光合源不足条件下,测定分析植株源与库的关系,应用同位素示踪技术分析连作地黄光合产物的分配及内源激素的变化,以探讨地黄的连作障碍机制。结果表明:剪叶处理对植株的影响不能达到连作障碍程度,其库容及其活性大于连作地黄;连作地黄出现光合产物滞留于地上部,库的活性影响了植株光合产物向库的运输,在生长前期植株内源激素发生了变化;因此断定,连作地黄库容及其活性过低是限制地黄块根膨大的重要因素,库通过不同激素对源的反馈调节,抑制了地上源的光合能力及光合产物向库的分配,导致连作障碍。     

The activities of antioxidant enzymes in response to oxidative stresses and hormones in paraquat-tolerant Rehmannia glutinosa plants

All members of R. glutinosa show the unique characteristic of intrinsic tolerance to paraquat (PQ). Antioxidant enzymes have been proposed to be the primary mechanism of PQ resistance in several plant species. Therefore, the antioxidant enzyme systems of R. glutinosa were evaluated by comparatively analyzing cellular antioxidant enzyme levels, and their responses of oxidative stresses and hormones. The levels of ascorbate peroxidase (APX), glutathione reductase (GR), non-specific peroxidase (POX), and superoxide dismutase (SOD) were 7.3-, 4.9-, 2.7- and 1.6-fold higher in PQ-tolerant R. glutinosa than in PQ-susceptible soybeans. However, the activity of catalase (CAT) was about 12-fold higher in the soybeans. The activities of antioxidant enzymes reduced after PQ treatment in the two species, with the exception of POX and SOD in R. glutinosa, which increased by about 40 %. Interestingly, the activities of APX, SOD and POX in R. glutinosa, relative to those in soybeans, were further increased by 49, 67 and 93 % after PQ treatment. The considerably higher intrinsic levels, and increases in the relative activities of antioxidant enzymes in R. glutinosa under oxidative stress support the possible role of these enzymes in the PQ tolerance of R. glutinosa. However, the relatively lower levels of SOD versus PQ tolerance, and the mixed responses of antioxidant enzymes to stresses and hormones, suggest a possible alternative mechanism(s) for PQ tolerance in R. glutinosa.     

地黄对芝麻的化感作用及其化感物质的分离鉴定

通过分离地黄(Rehmannia glutinosa)组织中抑制芝麻(Sesamum indicum)生长发育的化感物质, 以弄清地黄茬种植芝麻造成芝麻严重减产的原因。以芝麻为受体的生物测定为检测跟踪手段, 通过浸提、萃取、硅胶柱层析分离方法及气相色谱-质谱(GC-MS)鉴定方法, 分离鉴定了地黄组织影响芝麻的化感物质。对地黄不同组织的浸提液进行的生物测定结果表明: 影响芝麻生长发育的化感物质在地黄叶中含量高。地黄叶浸提液的不同有机相萃取组分的生物测定结果表明: 乙酸乙酯相化感活性最强, 乙酯乙酯相通过硅胶柱层析分离获得的洗脱组分Fr7化感活性最强。通过GC-MS及验证试验证明: 地黄抑制芝麻生长的化感物质是月桂酸和2,6-二叔丁基苯酚。研究表明地黄他感作用与自毒作用的化感物质是不同的。     

不同加工方法丹参花茶的质量评价

目的：研究丹参非传统药用部位花的制茶工艺,以及不同加工条件下丹参花茶的主要化学成分分析和感官评价。 方法：于盛花期采摘紫花丹参花蕾和开放初花,采用正交试验设计不同制茶工艺,采用紫外分光光度计测量制得花茶中叶绿素、类胡萝卜素、可溶性糖、总多酚、氨基酸以及总黄酮含量,进一步采用HPLC法检测丹参花茶中迷迭香酸、丹酚酸B、丹参素的含量,并进行了相应的方法学考察。结果：丹参花茶中含有较高的总多酚,具体成分以迷迭香酸为主,含有少量的游离氨基酸、黄酮、多糖以及微量的叶绿素和类胡萝卜素。结论：采摘丹参花蕾利用微波杀青、微波干燥,可以保留较多活性成分,同时具有较好的外观和口感。     

地黄育种研究进展

地黄是一种玄参科药用植物,是我国著名的"四大怀药"之一,具有重要的药用价值和经济价值。该文综述了地黄的引种、选择育种、杂交育种、组织培养辅助育种、转基因育种和DNA分子标记辅助育种等育种研究的成果,以期为地黄育种工作者提供参考。     

地黄属种间亲缘关系研究

对地黄属6个物种进行了形态解剖学观察、染色体计数、核核糖体内转录间隔区(ITS)序列分析.结果表明,茎的高度、幼叶形态、花萼和花冠形态及颜色、种子千粒重和大小、外种皮网壁厚度、外种皮内侧网纹直径等均是该属内分类的可靠依据.高地黄与裂叶地黄在外部形态及解剖结构的各个方面均极为近似,地黄与茄叶地黄间也存在较大的相似性.茄叶地黄、高地黄、湖北地黄、天目地黄的染色体数目,分别为n=28、14、14、14,确认地黄和茄叶地黄为属内四倍体物种,其余种均为二倍体.ITS测序分析显示,地黄属为单系起源,天目地黄与湖北地黄、高地黄与裂叶地黄、地黄与茄叶地黄构成属内3个分支,与形态学及细胞学研究结果一致.研究认为,天目地黄与湖北地黄有较近的亲缘关系;高地黄和裂叶地黄应为同一物种;地黄与茄叶地黄是属内进化水平最高的类群.     

地黄连作对根际微生物区系及土壤酶活性的影响

以地黄连作2年和1年的土壤为研究对象,分别测定了根际微生物区系变化及根际土壤酶活性．结果表明：地黄连作对其根际微生物区系及土壤酶活性产生了较大的影响．随种植年限的增加,根际细菌和真菌减少,但差异均不显著;放线菌增多,连作2年的土壤约为1年的4倍．土壤中氨化细菌、好气性固氮菌、硫化细菌、反硝化细菌和嫌气性纤维素分解菌分别增加了25．99、45．39、11．43、1．36和1．43倍,而好气性纤维素分解菌减少了86．74％．连作地黄根系的分泌物对脲酶、多酚氧化酶、蔗糖酶、蛋白酶和纤维素酶活性具有促进作用,分别增加了62．87％、9．43％、47．91％、139．62％和31．33％,而对过氧化氢酶则呈抑制作用．说明地黄连作会破坏根际微生物种群平衡．     

响应内生菌侵染的两个地黄茉莉酸合成关键基因的克隆与表达分析

丙二烯氧化物合酶(allene oxide synthase,AOS)、12-氧代植二烯酸还原酶(12-oxophytodienoate reduc?tase,OPR)基因是植物中茉莉酸合成关键基因。从地黄与内生真菌GG22互作转录组中筛选响应内生菌侵染的AOS和OPR基因序列,设计特异引物,克隆RgAOS和RgOPR基因的完整开放阅读框,并对该序列及其编码产物进行生物信息学分析,采用实时荧光定量PCR技术分析RgAOS和RgOPR基因在不同组织及内生真菌GG22侵染前后的表达模式。RgAOS基因开放阅读框为1626 bp,编码541个氨基酸,分子量为60.20 kD。RgOPR基因的开放阅读框为1197 bp,编码398个氨基酸,分子量为44.07 kD。QPCR分析发现,RgAOS在根中表达量最高,花中最少,RgOPR在花和叶中的表达量较高。内生真菌GG22能诱导地黄中RgAOS和RgOPR基因的表达。该研究成功从地黄中克隆了RgAOS和RgOPR基因,为进一步研究地黄中茉莉酸类物质的生物活性及探索内生真菌对地黄次生代谢产物调节的分子机制奠定了基础。     

怀地黄叶片中总环烯醚萜苷及总苯乙醇苷含量快速分析模型的建立

本文旨在建立地黄叶片中总环烯醚萜苷及苯乙醇苷定量分析模型。利用紫外-可见分光光度法测定不同种质怀地黄生育期内的128份地黄叶片中总环烯醚萜苷及总苯乙醇苷的含量,并将其作为基础值,结合地黄叶片的近红外光谱图,利用TQ8.0分析软件结合偏最小二乘法(PLS),分别建立地黄叶片中总环烯醚萜苷及总苯乙醇苷的定量分析模型。地黄叶片中总苯乙醇苷定量校正模型决定系数(R2)为0.998 2,校正均方根偏差(RMSEC)为0.089 9,预测均方决定差(RMSEP)为0.142,交叉验证均方根偏差(RMSECV)为0.707 2;总环烯醚萜苷定量校正模型的内部交叉验证决定系数(R2)为0.972 1,校正均方差(RMSEC)为0.259,预测均方决定差(RMSEP)为0.095 4,交叉验证均方根偏差(RMSECV)为0.869 4。预测值与实测值差异无统计学意义。该定量模型可用于怀地黄叶片中总环烯醚萜苷及总苯乙醇苷含量的快速测定。     

Silencing of the N family of resistance genes in Nicotiana edwardsonii compromises the hypersensitive response to tombusviruses

The nontarget effects associated with silencing of the N gene in Nicotiana edwardsonii, an amphidiploid species derived from N. glutinosa and N. clevelandii, have been characterized in this study. The N protein confers resistance to Tobacco mosaic virus (TMV), and is representative of a family of nucleotide-binding site leucine-rich repeat proteins present in N. glutinosa. Previous studies have shown that silencing of the N gene or of other plant genes associated with N-mediated defenses abolishes host resistance to TMV, and this effect can be measured through enhancements in movement or replication of TMV in the N-silenced plants. However, the nontarget effects of gene silencing have not been investigated thoroughly. Notably, are the functions of other resistance (R) genes also affected in experiments designed to silence the N gene? To investigate whether heterologous sequences could silence the N gene, we selected an R gene homolog from N. glutinosa that differed from the N gene by approximately 17%, created a hairpin transgene, and developed transgenic N. edwardsonii plants. Expression of this hairpin in the transgenic N. edwardsonii plants compromised the hypersensitive response to TMV, demonstrating that a single hairpin transgene could silence a block of R genes related by sequence similarity. We then investigated whether the response of N-silenced plants to other viruses would be altered, and found that the hypersensitive response triggered against the tombusviruses Tomato bushy stunt virus and Cymbidium ringspot virus also was compromised. This study indicates that a Tombusvirus R gene shares some homology with the N gene, which could facilitate the cloning of this gene.     

Chloroplast DNA sequencing and detailed microsatellite genotyping of all remnant populations suggests that only a single genet survives of the critically endangered plant Rehmannia japonica

Journal of Plant Research - Rehmannia japonica (Thunb.) Makino ex T. Yamaz. is an endangered perennial herb species in Japan. Although earlier the Japanese considered it a variety of R. glutinosa,...     

地黄属和崖白菜属等位酶的变异研究

利用超薄平板微型聚丙烯酰胺凝胶的等电聚焦电泳技术,对玄参科Scrophulariaceae地黄属Rehmannia 6种植物和崖白菜属Triaenophora 2种植物共52份材料进行了7个酶系统的等位酶分析。在7个酶系中共检测到18个清晰位点和45个等位基因,多态位点为17个;并将遗传学数据用软件Biosys-1进行分析,结果表明:同种内个体间表现出较大的遗传同质性,而不同种的个体则表现出明显的趋异性。除高地黄R.elata和裂叶地黄R.piasezkii外,其余6种植物之间遗传分化较明显,8个种按聚类结果分为两支,崖白菜属的崖白菜T.rupestris和神农架崖白菜T.shennongjiaensis聚为一支,位于分支树的基部;地黄属的地黄R.glutinosa、茄叶地黄R.solanifolia、天目地黄R.chingii、湖北地黄R.henryi、高地黄和裂叶地黄为另一支。在地黄属中,高地黄和裂叶地黄之间的亲缘关系最近,地黄和裂叶地黄关系最远。本研究结果不支持把高地黄和裂叶地黄作为两个独立的种。地黄属与崖白菜属在遗传上具有较高水平的相似性,它们在生活史和形态学上仍然保留有许多共同的特征,说明它们之间分化的时间可能不长。     

Early interactions between Alnus glutinosa and Frankia strain ArI3. Production and specificity of root hair deformation factor(s)

Actinomycetes from the genus Frankia are able to form symbiotic associations with more than 200 different species of woody angiosperms, so called actinorhizal plants. Many actinorhizal plants are infected via deformed root hairs. Factor(s) eliciting root hair deformation in actinorhizal symbioses have been found to be released into the culture medium, but the factor(s) has (have) not yet been characterized. In the present work, we describe the constitutive production of factor(s) by Frankia strain ArI3 causing root hair deformation on Alnus glutinosa . Deformation was detected after 4–5 h of incubation with both Frankia cultures and their cell-free culture filtrates. When culture filtrate was used, deformation was concentration dependent. A contact time of 2 min between culture filtrate and host roots was sufficient to induce subsequent root hair deformation. No root hair deformation on A. glutinosa could be detected with purified Nod factors from Rhizobium meliloti or R. leguminosarum biovar viciae . No correlation was found between Frankia strains belonging to different host specificity groups and their ability to deform root hairs on A. glutinosa. However, strains not able to deform root hairs on A. glutinosa were also unable to nodulate.     

Two new eudesmane alcohols from Jasonia glutinosa

Two new sesquiterpene alcohols have been isolated from the aerial parts of Jasonia glutinosa D. C. The structure of these sesquiterpenes were characterized by 1D and 2D NMR techniques (DQCOSY. TOCSY, NOESY, HMQC and HMBC) as (11R)-eudesm-4-en-11,12-diol and (11R)-eudesmane-5alpha, 11,12-triol.     

Calcium paradox from cyclostome to man: a comparative study

In seven poikilotherm species, i.e. cyclostome, Myxine glutinosa; teleosts, Gadus morrhua; Platichtys flesus; amphibia, R. pipiens; R. temporaria; Xenopus laevis; reptiles, Vipera berus. Ca paradox could not be obtained in ventricular strips at 8-12 degrees C. In similar preparations from human atria at 30 degrees C and 37 degrees, Ca paradox was obtained similarly as in other homoiotherms. In homoiotherms, lowered temperature (below 30 degrees C) has a protective effect. Viper myocardium did not display Ca paradox at 12, 22 or 32 degrees C, whereas human myocardium displayed Ca paradox even at 30 C. It is postulated that the absence of Ca paradox in poikilotherms is not due to the low temperature. In hagfish (Myxine glutinosa) the absence of Ca paradox seems to be due to the specific conditions concerning Ca exchange in the myocardial cell (large glycocalyx).     

地黄连作的生理生态特性

选择具有典型连作障碍效应的药用植物地黄(Rehmannia glutinosa)为试验材料, 以正茬地黄为对照, 研究连作条件下地黄植株的生理生态特性变化。结果表明, 该试验条件下地黄的连作障碍效应起始于苗期, 在栽种后60天时, 连作引起植株细胞膜质过氧化作用, 使膜的正常结构和功能受到损伤; 重茬地黄的光合速率(P_n)、气孔导度(G_s)和胞间CO₂浓度(C_i)显著低于对照, 而气孔限制值(L_S) 33.97%, 显著高于对照, 同期重茬地黄叶绿素含量(Chl)也开始显著低于对照。相关分析表明, P_n与G_s、Chl含量相关(相关系数分别为0.977和0.814)。对植株叶片叶肉细胞的电镜观察显示, 连作条件下地黄光合细胞结构在其生长中期开始发生变化。因此, 在地黄生长早期由于(连作)环境胁迫植株叶片细胞内活性氧、自由基积累所带来的膜结构损伤, 导致了叶绿素含量的降低, 而气孔关闭以及叶绿素含量的降低共同造成连作地黄早期光合能力降低, 生长受阻, 从而表现出明显的障碍效应。     

地黄叶和茎的解剖学及组织化学研究

采用解剖学和组织化学方法对地黄叶和茎的显微结构以及梓醇、多糖的分布进行观察研究,以明确梓醇和多糖在地黄叶和茎中的分布特征。结果显示:(1)地黄叶的上、下表皮均分布有腺毛和非腺毛,腺毛都属于头状腺毛,包括长柄和短柄的头状腺毛,两类腺毛的分泌物化学成分主要是黄酮和多糖;叶的上、下表皮上都分布有无规则型气孔,下表皮的气孔密度比上表皮的大,但气孔指数相差不大;栅栏组织由2～3层薄壁细胞构成,排列紧密,海绵组织薄壁细胞形状无规则,细胞间隙大。(2)组织化学研究表明,海绵组织中黄斑样的薄壁细胞是梓醇和多糖的贮存场所,这类薄壁细胞在叶片边缘的齿末端处最为集中,茎的皮层、韧皮部和木质部的薄壁细胞也都是梓醇和多糖的贮存场所。     

黑长臂猿栖息地旱冬瓜和潺槁木姜子种群分布格局和动态

旱冬瓜（Alnus nepalensis）和潺槁木姜子（Litsea glutinosa）是云南省无量山黑长臂猿（Nomascus concolor）栖息地中的两种常见乔木.利用最近邻体法和Heygi单木竞争指数模型,对两个种群的分布格局和竞争情况进行分析.结果表明,旱冬瓜在不同生长阶段均呈现随机分布,而伴生的潺槁木姜子种群则呈现随机分布 聚集分布 均匀分布的变化趋势.种间竞争和物种自身生物学特性对两个种群的分布格局具有显著影响.旱冬瓜种群的龄级结构为衰退型,潺槁木姜子种群则为成长型.     

地黄自毒物质提取及其生物指标测定

连作障碍在药用植物地黄(Rehmannia glutinosa L.)栽培中尤其明显,同一地块上种植一茬地黄后,须经8-10a后方可再种。从怀地黄主产区河南省焦作地区采集种植1a地黄的茬后土壤,采用不同溶剂提取地黄土壤自毒物质并利用生物测试确定抑制率最强的土壤提取液作温室栽培实验以确定对植物生理指标的影响。利用高效液相色谱法(HPLC)对土壤提取液和根系分泌物进行比较,并利用高效液相色谱、傅立叶转换红外光谱(FTIR)和电喷雾离子阱质谱(ESI-MS)技术鉴定地黄连作自毒物质的化学成分。生物测试结果表明水和甲醇提取的地黄茬后土壤自毒物质对地黄生长具有最强的抑制作用;在培养器中加入浓度0.5g/mL的水和甲醇提取液对地黄胚根的抑制率分别达17%和26%,当浓度增至5.0g/mL时,抑制率增加到70%以上。盆栽结果显示甲醇提取的自毒物质导致地黄地上部叶绿素含量减少,根系活力下降,明显抑制了地黄体内超氧化物歧化酶、过氧化物歧化酶、过氧化物酶的活性,加剧了地黄膜质过氧化,以及引起地黄植体内生长素类激素含量下降。高效液相色谱法显示土壤提取液和根系分泌物具有相似图谱。电喷雾离子阱质谱分析共检测到6个特征物质:香草酸、D-甘露醇、二十六烷酸苯羟基乙酯、毛蕊花糖苷、β-谷甾醇和胡萝卜苷。     

The evolution of fraction I protein during the origin of a new species ofNicotiana

The polypeptide composition of Fraction I protein from Nicotiana digluta, a synthetic species which arose by chromosome doubling following the interspecific hybridization of N. glutinosa and N. tabacum, has been examined by isoelectric focusing. The composition of the protein from N. digluta, which was identical to the protein from the infertile F1 hybrid N. glutinosa x N. tabacum, showed 3 polypeptides in the large subunit and 4 polypeptides in the small subunit. The large subunit polypeptides were identical to those from N. glutinosa, the maternal parent in the original hybridization, whereas the small subunit polypeptides were a composite of the small subunit polypeptides from both N. glutinosa and N. tabacum. This analysis demonstrates how the polypeptide composition of Fraction I protein evolves during the origin of new species of Nicotiana.