梁山古莲(Nelumbo nucifera Gaertn.)的种质资源价值研究

对2013年梁山出土的古莲(命名为梁山古莲)进行了种质资源价值研究。断代分析结果显示,梁山古莲籽距今时间最长约为731 a,入土年代处于我国元朝初期,古莲籽发芽率在90%以上,不同方位取得的莲籽大小存在明显差异;表型分析表明,梁山古莲属于中国莲系大株型少瓣红莲类群,为黄河流域野生莲,古莲群体存在较高的遗传多样性;聚类分析显示,梁山古莲与山东地方品种马踏湖白莲亲缘关系最近。

Genetic Variation in Rhizome Lotus(Nelumbo nucifera Gaertn.ssp.nucifera) Germplasms from China Assessed by RAPD Markers

To estimate genetic variation in rhizome lotus （Nelumbo nucifera Gaertn. ssp. nucifera） germplasms in China, a total of 94 rhizome lotus germplasms collected from 18 provinces in China were assessed. The RAPD （randomly amplified polymorphic DNA） marker was employed. The selected 17 random primers detected 139 polymorphic alleles out of a total 207 （67.15%）. Nei＇s gene diversity statistics and region differentiation parameters indicated that all germplasms had a relatively high level of genetic diversity with ne = 1.3202, h = 0.1937, I= 0.2982 and the gene flow among all regions was Nrn = 5.5742. The UPGMA dendrogram clustered all 94 germplasms into two clusters： One contained eight commercial cultivars and major landraces, and the other included the wild and some special landraces from five regions, and the PCA analysis exhibited the similar result. Those germplasms from southwestern and eastern China had higher genetic diversity than those from the southern, northern and central China. Predominant proportion of genetic variation （95.61%） was found significant within rather than among （4.39%） regions, as revealed by AMOVA analysis. The data analysis also revealed that the genetic diversity of rhizome lotus germplasms among different regions is positively related to their geographic distances, though it is ambiguous to find the trend from the UPGMA dendrogram and the PCA analysis. A relatively high genetic diversity and gene flow resided in the root lotus germplasms; about 96% of the variation was found within region; accessions from southwest and eastern China have higher genetic diversity than those from the southern, northern and central China.

Genetic Diversity of Lotus(Nelumbo nucifera Gaertn.) Cultivars Revealed by ISSR Analysis

[ Objectives ] This study was conducted to investigate the genetic diversity of 20 lotus （Nelumbo nucifera Gaertn） samples. [ Methods ] On this optimal ISSR amplification system, 16 primers were screened with good polymorphism, and the DNA was used to amplify the 20 plant samples. [ R^ults] The 16 primers produced 225 loci, of which 170 were polymorphic, and the polymorphic loci percentage was up to 75.56%. The genetic similarity coefficients between the 20 vari- eties ranged from 0. 577 8 to 0.951 1, which were calculated by POPGENE32. The 20 varieties by UPMGA analysis could be clustered into 2 groups, the first of which included Baiyangdian red lotus and Donggua lotus, and other varieties was included in the second group. [ Conchmions] ISSR molecular markers could be effectively used in genetic diversity and fingerprint analysis for different lotus varieties.

中国莲(Nelumbo nucifera)幼苗抗氧化系统对砷胁迫的响应

为了研究中国莲(Nelumbo nucifera)抗氧化系统对砷胁迫的响应,研究比较了两种不同价态无机砷As(Ⅲ)和砷As(Ⅴ)对中国莲幼苗可溶性蛋白含量、丙二醛(MDA)含量以及抗氧化系统的影响。结果表明,中国莲幼苗中MDA和可溶性蛋白质含量随砷浓度的增加呈现先升高后降低的趋势。中国莲幼苗的MDA和蛋白质含量受As(Ⅴ)的影响不如As(Ⅲ)敏感。抗氧化系统酶中,超氧化物歧化酶(SOD)对砷处理最敏感,当As(Ⅲ)浓度在2.5μmol/L和As(Ⅴ)浓度在100μmol/L时,SOD酶活性显著高于对照组。过氧化物酶(POD)在As(Ⅲ)处理浓度为10μmol/L时就出现显著上升,相对而言,过氧化氢酶(CAT)对As(Ⅴ)比较敏感。实验结果表明,随着浓度的增加,砷对幼苗产生的氧化胁迫导致SOD、CAT和POD三种酶活性有所增加,以配合清除细胞内的活性氧自由基(ROS),维持细胞代谢的稳定。本研究为进一步研究砷胁迫下莲的生理和生长变化、以及莲的培育和移植提供了部分基础数据。

A new benzylisoquinoline alkaloid from stems of Nelumbo nucifera

A new benzylisoquinoline alkaloid, named nelumstemine （1）, 1-（4＇-hydroxybenzoyl）-6,7-dimethoxy-3,4-dihydroisoquinoline, was isolated from the stems of Nelumbo nucifera Geartn. Its structure was established on the basis of spectral analysis.

Analysis of Accumulating Ability of Heavy Metals in Lotus (Nelumbo nucifera) Improved by Ion Implantation

Heavy metals have seriously contaminated soil and water, and done harm to public health. Academician WANG Naiyan proposed that ion-implantation technique should be exploited for environmental bioremediation by mutating and breeding plants or microbes. By implanting N^＋ into Taikonglian No.l, we have selected and bred two lotus cultivars, Jingguang No.1 and Jingguang No.2. The present study aims at analyzing the feasibility that irradiation can be used for remediation of soil and water from heavy metals. Compared with parent Taikonglian No.l, the uptaking and accumulating ability of heavy metals in two mutated cultivars was obviously improved. So ion implantation technique can indeed be used in bioremediation of heavy metals in soil and water, but it is hard to select and breed a cultivar which can remedy the soil and water from all the heavy metals.

Genetic diversity of the wild Asian lotus (Nelumbo nucifera) from Northern China

Asian lotus(Nelumbo nucifera Gaertn.)is an aquatic plant with ornamental,cultural,economic,and ecological values.China has abundant germplasm resources of Asian lotus.However,in many areas,the wild Asian lotuses have been destroyed and the germplasms is now facing extinction.In addition,the knowledge of the genetic diversity of the wild Asian lotus in China is poor.To identify and protect the germplasms of Chinese wild Asian lotus,eleven genomic-SSR primers,three EST-SSR primers,and three chloroplast DNA primers were used to investigate the genetic diversity among 69 samples of wild Asian lotus from 25 locations in northern China.The genetic diversity of 27 samples of wild Asian lotus from southern China and other countries,the ancient Asian lotus,Asian lotus cultivars from China,and Asian-American hybrids was also compared.The genetic diversity of the wild Asian lotus from northern China was characterized as medium,and themean values of observed heterozygosity(Ho)and expected heterozygosity(He)were 0.087 and 0.552,respectively.Based on a UPGMA dendrogram and STRUCTURE analysis,the wild Asian lotus samples in northern China were divided into three groups.The wild Asian lotus samples from northern China contained 16 haplotypes.The Nei’s genetic distance between the wild Asian lotus samples from the Songhua River basin and the Liao River basin in northeastern China was relatively small,and these germplasms might be relatively primitive compared to those from other regions.This study provides essential information regarding the genetic diversity of the wild Asian lotus resources in northern China,and provides a basis for further analysis of population-level genetic evolution through high-throughput sequencing.

Gibberellins,brassinolide,and ethylene signaling were involved in flower differentiation and development in Nelumbo nucifera

Hormones play important roles in vegetative and reproductive processes;however,the regulatory roles of hormones in Nelumbo nucifera(Lotus)growth and development are unclear.In this study,nine types of endogenous hormones,including gibberellins(GA_1,GA_3,and GA_4),indole-3-acetic acid(IAA),brassinolide(BR),ethylene(ETH),jasmonic acid(JA),abscisic acid(ABA),and zeatin(ZT)were detected in dormant shoot tips,vegetative shoot tips,developing leaf buds,and developing flower buds of lotus.The results indicated that GA,ETH,and BR signaling can promote vegetative and reproductive development of lotus.GA signaling regulates plant height and stimulates flower bud differentiation.GA levels were the highest in the flower buds;exogenous GA3+4 increased plant height by approximately 90%,increased flower quantity by nearly 40%,and advanced flowering by 4 d.Suppressing GA biosynthesis using paclobutrazol decreased plant height and flower quantity by 38%and 87.1%,respectively,and delayed flowering by 15.6 d.ETH signaling has positive regulatory effects on vegetative growth and flower development.The ETH concentration in the developing leaf buds was at least 50%higher than that in other samples.Ethephon spraying led to remarkable increases in plant height and leaf thickness and extended the flowering duration.BR signaling acts as a growth promoter during vegetative and reproductive development in lotus.The highest BR levels were detected in the vegetative shoot tips.External application of28-epihomobrassinolide resulted in growth-promoting phenotypes including longer scapes,thicker leaves,and prolonged flowering.

采用AFLP技术分析17个荷花(Nelumbo nucifera)品种的系统关系

【目的】采用AFLP技术,对17个荷花品种进行亲缘关系分析。【方法】从64个AFLP引物组合中筛选出10对多态性较高的引物,以17个荷花DNA为模板,进行PCR扩增,并对产物进行聚丙烯凝胶电泳分析。【结果】共获得条带523条,其中多态性条带370条,多态性比例达到70.75%;根据扩增条带信息进行聚类,可将17个品种分为2组;其中有5对引物,每对引物单独使用均可将17个品种完全区分开。【结论】AFLP用于鉴别荷花品种,具有较高的可行性;本研究为荷花分类和杂交育种的亲本选择提供参考信息。

Evaluation of the Anti-Hyperlipidemic Activity of <i>Nelumbo nucifera </i>Fruit in Rabbits Fed with High Cholesterol Diet

Recently use of herbal therapies and diet rich in flavonoids and vitamins has augmented significantly to manage minor to modest illnesses. Several anti-hyperlipidemic agents are currently available;however most of them have various unwanted effects. That’s why more people are switching towards safer alternatives, specially derived from plants with fewer side effects. Hence current study was focused to determine the anti-hyperlipidemic potential of Nelumbo nucifera fruit in rabbits fed with high cholesterol diet. This study would surely help in the exploration of a novel agent from the plant source and therefore may recommend the application of traditional medicines in humans. 28 white healthy rabbits of either sex weighing 1000 - 1500 gm were divided into four groups. Control group was given gum tragacanth and 3 groups served as test groups and were given N. nucifera fruit extract 50, 100 and 200 mg/kg for 45 days. Initially animals of all four groups received high cholesterol diet (HCD) 0.125 gm/kg cholesterol in 0.5% corn oil for 30 days. All drugs were given orally. Blood samples were taken thrice from the ear vein of animals, 1st after 24 hours of thirty days of HCD then again after 24 hours of thirty and forty five days dosing of N. nucifera fruit extract. The results showed remarkable cholesterol lowering effects of N. nucifera fruit extract at doses 50, 100 and 200 mg/kg but were particularly dominant at 200 mg/kg at which fruit extract highly significantly reduced TC, TG and LDL-C after 30 and 45 days and increased HDL-C significantly after 30 days and highly significantly after 45 days. Flavonoids, saponins, tannins and procyanidin are significant constituents of N. nucifera fruit which may have contributed in lipid lowering effects of N. nucifera fruit which can contribute in lowering the risk of CVDs and stroke but more preclinical and clinical studies are required to confirm these findings.

Genetic Variation of the Novel Badnaviruses Infecting Nelumbo Nucifera Based on the RT/RNase H Coding Region Sequences

Lotus(Nelumbo nucifera Gaertn.)is an economically important perennial aquatic herb plant in China.Presently,viral diseases caused by Cucumber mosaic virus(CMV)and Dasheen mosaic virus(DsMV)are detected in lotus.In this study,the putatively novel badnaviruses from lotus plants(LBVs)were identified using next-generation sequencing of siRNAs and conventional Sanger sequencing based on the RT/RNase H coding region sequences.Lotus plants infected by LBVs harbored virus quasispecies.A unique recombination event of LBVs was found in a single lotus plant.Sap inoculation showed that LBVs could actively replicate in the lotus plants,but without a wide host range.The field survey of 43 lotus plants in Jiangsu Province showed a prevalence of 62.8%for LBVs,confirming that it is widely distributed in Jiangsu Province of China.

莲株型调控的研究进展

株型是园林植物的重要观赏性状,培育丰富的观赏性状是园林植物的育种目标之一。莲作为我国重要的园林植物,其理想株型育种应向高大型和极微型2个极端株型发展。本研究对莲株型调控的细胞学基础、生理机制、分子机理进行了系统地综述,提出了深入探究茎顶端分生组织、居间分生组织的发育模式及基因调控机理,挖掘GAs、AUX在合成和信号传导过程中的关键基因、利用分子标记技术开发莲株型相QTL位点和相关基因精准定位的未来研究方向。

不同品种荷花耐盐碱性评价及鉴定指标筛选

为研究不同荷花(Nelumbo nucifera)品种在混合盐碱胁迫下的耐受程度,筛选抗性品种,采用盆栽法,以20个荷花品种为试验材料,对其进行100 mmol·L^(-1)(NaCl∶NaHCO3=2∶1)混合盐碱胁迫处理,7 d后测量株高、叶面积、光合色素、叶绿素荧光参数、丙二醛含量、抗氧化物酶活性及渗透调节物质含量,通过耐盐碱系数、相关性分析、主成分分析、隶属函数法、聚类分析等方法对20种荷花耐盐碱性进行综合评价。结果表明:不同荷花品种单项指标的耐盐碱系数变异系数差异较大,除超氧化物歧化酶活性和过氧化物酶活性外,其余单项指标的耐盐碱系数均存在显著或极显著相关性,利用主成分分析将12个单项指标转换为5个综合指标,各综合指标贡献率分别为36.175%、18.709%、13.470%、11.379%、6.610%,累计贡献率达到86.344%。以耐盐碱性综合评价值(D值)为依据进行聚类分析,将20个荷花品种分为4类,第一类为强耐盐碱型,共计4个荷花品种,第二类为耐盐碱型,共计5个荷花品种,第三类为不耐盐碱型,共计8个荷花品种,第四类为敏感型,共计3个荷花品种。通过逐步回归分析建立荷花的耐盐碱性最优评价模型,D=0.453+0.148X_(1)+0.122X_(2)-0.122X_(7)-0.033X_(8)+0.093X_(11)+0.037X_(12)(R^(2)=0.992),估计精度95.45%以上,并筛选出株高、叶面积、丙二醛含量、脯氨酸含量、可溶性糖含量和可溶性蛋白含量为荷花耐盐碱性评价指标,可以在相同的条件下,通过测定这6项指标,确定荷花的耐盐碱性强弱。

湘莲根际促生菌的筛选及其效应研究

为获得湘莲根际促生菌(Plantgrowth promoting rhizobacteria,PGPR)并明确其促生特性,试验从湘莲根际土壤中分离出7株固氮解磷菌,通过对其功能指标的对比挑选出菌株XL7,其具有良好的固氮解磷和产IAA能力。经鉴定该菌株为高地芽孢杆菌(Bacillus altitudinis)。通过种子萌发、盆栽试验测定菌株对湘莲的促生效应。结果表明:接种XL7能促进种子的萌发,发芽势、发芽率、苗长、苗重和不定根重均高于对照;土壤中矿质态氮、速效磷和速效钾分别提高了5.3%、31.8%和26.1%;莲幼苗植株生物量、全氮、全磷和全钾分别增加了25.0%、18.3%、80.5%和38.7%。这表明该多功能菌株对湘莲具有良好的促生作用。

山东莲藕(Nelumbo nucifera Gaertn.)种质资源的RAPD鉴定和遗传多样性分析

山东莲藕(Nelumbo nucifera Gaertn.)栽培历史悠久,在中国北方具有重要影响。但是,山东莲藕种质资源研究一直没有开展。为收集和保存莲藕种质资源,本研究从山东省8个种植面积较大的地区采集了76份莲藕材料,并利用RAPD分子标记对这些材料进行了遗传相似性分析,共鉴定出38份种质资源。遗传多样性指数分析显示,38份种质资源的Nei's多样性指数(H)平均值为0.26,Shannon信息指数(I)平均值为0.41。研究数据表明,山东莲藕种质资源相对丰富,但要建立高质量的种质资源圃,仍需要加强种质资源的引进。

莲藕酚类物质组成分析及合成机制研究

目的分析莲藕采前不同时期酚类物质的组成,筛选莲藕特征酚类物质代谢相关基因及转录因子。方法本研究以“鄂莲五号”为试材,基于高效液相色谱及转录组学解析莲藕采前特征酚类物质代谢及转录组调控相关基因。结果不同生长期莲藕褐变度呈上升趋势,其总酚含量先增加后减少,S3(生长期3)时期总酚含量达到最高峰为16.59 mg GAE/100 g。其中酚类物质含量最高的是天麻素,在S4时期含量达到最高峰为191.44μg/g。苯丙烷和类黄酮代谢途径中参与天麻素、没食子儿茶素、儿茶素合成的基因在莲藕根茎膨大早期和中期高表达。利用实时荧光定量聚合酶链式反应验证转录组分析检测到的酚类化合物生物合成所需的几个差异表达基因。27个转录因子家族在莲藕生长期显著差异表达,其中Nn_104608167(AP2-EREBP)、Nn_104608008(MYB)、Nn_104593222(bHLH)、Nn_104594756(C2C2-Dof)表达变化与生长期莲藕酚类物质含量变化模式相一致。结论莲藕生长期酚类物质的不断积累,导致褐变度呈现出不断上升趋势。27个转录因子家族的部分基因表达量与生长期莲藕酚类物质变化趋势相一致,可能参与了酚类物质的转录调控。本实验为进一步研究莲藕酚类物质变化提供分子水平的理论基础。

莲藕多糖的碱法提取工艺优化与抗氧化活性评价

目的 优化莲藕多糖的碱法提取工艺并评价其抗氧化活性。方法 以多糖提取率为指标,以料液比、NaOH浓度、提取时间和提取温度为考察因素进行单因素实验,结合响应面法对提取工艺进行优化,并分析所得莲藕多糖的纯度、相对分子质量和自由基清除能力。结果 响应面优化结果显示,除料液比以外,NaOH浓度、提取时间和提取温度对莲藕多糖提取率均具有显著影响(P<0.05)。确定最佳工艺条件为:料液比1:16(g/mL)、NaOH浓度0.04 mol/L、提取时间1.3 h、提取温度57℃,此条件下的多糖提取率达13.07%,与预测值13.52%相近。该碱法提取多糖的纯度达73.66%,平均相对分子质量为1.727×10^(5)Da。抗氧化活性评价结果显示,莲藕多糖具有清除1,1-二苯基-2-三硝基苯自由基和羟基自由基的能力,且清除率与多糖浓度成正比。结论 优化了碱提莲藕多糖的提取工艺,且提取得到的多糖具有较好的抗氧化活性,为莲藕资源的开发利用提供了理论依据。

荷花(Nelumbo nucifera)全基因组PRR基因家族鉴定及其在多种非生物胁迫下的表达模式

为了探究荷花(Nelumbo nucifera)伪应答调控蛋白(pseudo-response regulators, PRR)NnPRR家族成员的特征及其在非生物胁迫响应中的作用,通过生物信息学分析,将7个NnPRR基因从荷花全基因组中挖掘出来,对其基因结构、蛋白保守基序、顺式作用元件及共线性关系等进行了鉴定分析,并利用实时定量PCR技术探究各基因分别在外源施加300 mmol/L NaCl、 5 mmol/L水杨酸(salicylic acid, SA)及4℃低温处理下的表达模式。研究结果表明,该家族基因在荷花中可分为3个分支,集中分布在chr2、chr3、chr5和chr6染色体上,该家族蛋白均定位于细胞核。该家族基因在启动子区域发现大量光响应元件,且一些基因还具有低温、茉莉酸甲酯及脱落酸等响应元件。从基因的表达模式上看,NnPRR基因家族7个基因均正向响应外源NaCl处理,而NnPRR2、NnPRR4、NnPRR3等基因则特异性响应外源SA或低温胁迫,推测该家族基因特异性参与荷花非生物逆境胁迫。本研究结果为深入研究荷花PRR基因家族响应逆境胁迫提供了理论基础,也为抗逆新品种的选育提供了新的基因资源。

荷叶中的生物碱及其对5-HT 2A和5-HT 2C受体的激动作用研究

对荷叶中的生物碱进行了分离、鉴定和调脂减肥活性研究。本研究结合传统酸提碱沉法与现代高效液相色谱制备技术,从荷叶中分离、纯化到11个生物碱,分别被鉴定为N-氧基原荷叶碱(1)、原荷叶碱(2)、莲碱(3)、降氧化北美黄连次碱(4)、荜茇宁(5)、巴婆碱(6)、O-去甲基荷叶碱(7)、N-去甲基荷叶碱(8)、荷叶碱(9)、衡州乌药碱(10)和亚美罂粟碱(11),其中,化合物1、4和5为首次从荷叶中分得。测试所得化合物对5-HT_(2A)和5-HT_(2C)受体的激动作用,结果表明11个生物碱对5-HT_(2A)受体均具有一定的激动作用,进一步揭示了荷叶调脂减肥的可能药效基础和作用机理。

不同观赏荷花品种对镉胁迫响应的差异性研究

[目的]本文旨在探讨不同观赏荷花品种耐镉(Cd)的差异,筛选优质荷花品种以作为植物修复Cd污染水体的材料。[方法]以常用的6个大中型观赏荷花品种为试材,利用盆栽试验研究不同Cd处理水平(0、1、10、25 mg·kg-1)下各荷花品种生长及生理特性的变化。[结果]低Cd处理(1 mg·kg-1)促进了各荷花品种株高和立叶数的增加,中Cd处理(10 mg·kg-1)对‘风卷红旗’‘中国红·北京’‘珠峰翠影’的株高和立叶数有促进作用,但抑制其他3个品种的生长,经高Cd处理(25 mg·kg-1)后仅‘珠峰翠影’的株高和立叶数仍显著增加;不同Cd处理水平对各荷花品种冠幅和叶径的生长均产生了抑制效果。随着Cd处理水平升高,各荷花品种叶片的叶绿素含量逐渐降低,相对电导率不断上升,超氧化物歧化酶(SOD)和过氧化物酶(POD)活性均呈先上升后下降的趋势。其中,‘风卷红旗’‘中国红·北京’和‘珠峰翠影’叶绿素含量及相对电导率受Cd胁迫的影响较其他3个品种小,且‘风卷红旗’和‘中国红·北京’的2种抗氧化酶活性仅在高Cd胁迫下才显著低于对照,而‘珠峰翠影’在各水平Cd处理下始终高于对照。对不同荷花品种Cd积累特性的分析表明,‘风卷红旗’和‘中国红·北京’的Cd积累量相对较高,而‘珠峰翠影’的Cd转运系数最高;荷花不同器官中Cd积累量由大到小表现为根、叶、根状茎。[结论]不同观赏荷花品种耐Cd能力差异较大,‘珠峰翠影’‘风卷红旗’‘中国红·北京’的耐Cd性相对较强,这3个耐性品种可通过根系阻挡与固定大量Cd或高效启动抗氧化系统清除Cd诱导产生过量活性氧的方式抵御Cd胁迫。