Isobutyl Amides from Pepper (Piper nigrum L.)

Four isobutyl amides were isolated from the fruits of white pepper (Piper nigrum L.) and identified to be N-isobutyl-13-(3,4-methylenedioxyphenyl)-2E,4E,12E-tridecatrienamide (3, guineensine), N-isobutyl-2E,4E,8Z-eicosatrienamide (5), N-isobutyl-2E,4E-octadecadienamide (6) and N-isobutyl-2E,4E-decadienamide (7, pellitorine).     

胡椒叶片基因组DNA提取方法比较

目的：研究建立胡椒叶片中提取高质量DNA的方法。方法：采用各种CTAB法和SDS法的改良方法,提取胡椒叶片中的总DNA,并对DNA进行紫外和电泳检测。结果：改良CTAB法4和5提取的DNA经电泳检测,有一条明亮主带,且无拖尾现象,样品槽无荧光出现,说明抽出的DNA纯度较高,一致性好;测定其OD260和OD280值,并计算其比值,OD260/OD280值在1.8-2.0之间,提取率在51.667-60.000μg/g之间,获得的基因组DNA质量高。结论：改良CTAB法4和法5可从胡椒幼叶中提取高质量DNA。     

利用辣椒疫霉培养滤液体外筛选胡椒抗瘟病无性系研究

在胡椒（Piper nigrum Linn．)茎尖丛生增殖技术的基础上,以印尼大叶种“Lampong Type”无菌实生苗作外植体源,利用辣椒疫霉(Phytophthora capsici)培养滤液对胡椒茎尖及其增殖形成的丛生芽进行体外选择。辣椒疫霉培养滤液的不同灭菌方法对辣椒疫霉培养滤液的毒性影响显著,过滤灭菌方式可以保持辣椒疫霉培养滤液的毒性,而高温高压灭菌方式则不能。随着辣椒疫霉培养滤液浓度的增加,茎尖和丛生芽的存活率和增殖率都在下降。在存活的茎尖或丛生芽培养中,一部分可正常增殖,其余的形成愈伤组织,或者保持生长停滞的休眠状态。在选择性培养基上继代培养2次后进行生根和移栽,利用离体叶片针刺接种法对温室条件下生长的移栽植株进行抗瘟病测定。以3次抗病检测均无明显症状的植株作为抗病株。随着辣椒疫霉培养滤液浓度的增加,得到的再生植株数量降低,但其中抗病株的比例提高。利用过滤灭菌方式加入选择性培养基的处理中,25％、50％和75％的辣椒疫霉培养滤液分别获得1株、4株和3株抗病株,分别占各处理再生植株总数的1．54％、20．00％和42．86％,共获得8株,占该组处理再生植株总数的8．70％。     

Sequencing of Coat Protein Gene of an isolate of Cucumber mosaic virus Infecting Black Pepper (Piper nigrum L) in India

The coat protein gene of an isolate of Cucumber mosaic virus (CMV) associated with stunted disease affecting black pepper plant was cloned and sequenced. The coat protein gene comprised of 657 nucleotides encoding a protein of 218 amino acids. Sequence analysis showed that the gene was most closely related to CMV infecting Egyptian henbane plant in India, the member of subgroup I of CMV. The amino acid sequence identity with members of subgroup I was 92-99% while that with members of subgroup II was 77–79%. On the basis of sequence homology, it is concluded that CMV infecting black pepper in India is a strain of CMV belonging to subgroup I.     

Structures and Syntheses of Two Phenolic Amides from Piper nigrum L.

Two phenolic amides were isolated from the fruits of white pepper (Piper nigrum L.) and identified to be N-trans-feruloyl tyramine (2a) and N-5-(4-hydroxyphenyl)-2E, 4E-pentadienoyl piperidine (6a) on the basis of chemical and specrtal evidence. Both compounds were synthesized.     

Occurrence of symptomless source of Piper yellow mottle virus in black pepper (Piper nigrum L.) varieties and a wild Piper species

Symptomless nature of Piper yellow mottle virus (PYMoV) infection in three varieties of black pepper (Piper nigrum) (Panniyur 1, Panniyur 5 and Panchami) and a wild species of Piper (Piper colubrinum) was confirmed by polymerase chain reaction (PCR) using PYMoV specific primers. The virus could be transmitted from these PYMoV-infected symptomless plants onto symptom producing black pepper cv. Karimunda through mealybug vector, Ferrisia virgata and by graft transmission. About 20–50% seedlings showed typical symptoms of the PYMoV at 30 days after mealybug inoculations while it was 75–94% at 90 days after inoculation. PCR test of the inoculated seedlings confirmed the presence of PYMoV in 50–64%, 76–100% and 80–100% of plants in 30, 60 and 90 days after inoculation, respectively. Similarly, 50–66%, 91–100% and 100% of graft-transmitted plants showed typical symptoms of the disease at 30, 60 and 90 days after grafting. PCR test of the graft-transmitted plants showed 100% PYMoV infection at 60 days after grafting. The results clearly demonstrated the existence of PYMoV-infected symptomless plants that can act as source for secondary spread of the virus in the field.     

In silico analyses of superoxide dismutases (SODs) of rice (Oryza sativa L.)

Superoxide dismutases (SODs), members of the metalloenzymes family are most effective intracellular enzymatic antioxidant in aerobic organisms. These enzymes provide the first line of defense in plants against the toxic effects of elevated levels of reactive oxygen species (ROS) generated during various environmental stresses. The availability of high-throughput computational tools has provided better opportunities to characterize the protein features and determine their function. In the present study an attempt was made to gain an insight into the structure and evolution of subunits of SODs (Cu-Zn, Mn and Fe SODs) of rice. The 3-Dimensional structures of SODs were modeled based on available X-ray crystal structures and further validated. The primary sequence, secondary and tertiary structure analysis revealed Mn and Fe SOD to be structurally homologous while Cu-Zn SOD is un-related to either of them. Comparative structural study also revealed former two were dominated by α-helices followed by β-strands in contrast; Cu-Zn SOD dominated by β-strands. Molecular phylogeny indicated a common evolutionary origin of Mn and Fe SOD while Cu-Zn SOD may have evolved separately.     

Isolation of RNA from blackcurrant (Ribes nigrum L.) fruit

Extraction of high-quality RNA from blackcurrant fruit has hitherto proved difficult, probably owing to high levels of phenolic and polysaccharide components in the berries. The procedure described here is a modification of one described for grape berries, and yields RNA suitable for in vitro translations, RNA blot analysis, and cDNA library construction.     

大叶种胡椒实生苗茎尖培养和合子胚培养研究

利用各种表面消毒方法对采自海南岛三个地区的胡椒大田植株的外植体进行消毒试验,由于内源性污染,除胡椒成熟种子外,其它各种大田外植体的表面消毒均未能成功。以胡椒成熟种子无菌萌发的实生苗茎尖作外植体,在1/2MS(MS或B5)+1.5mg/LBA+0～0.2mg/LIAA(或NAA)上可实现丛生芽增殖。茎尖水平或竖直接种方法显著影响茎尖的增殖;水平接种茎尖的生长和增殖效果优于竖直茎尖接种方式。茎尖增殖率随BA浓度的增加而提高,但BA浓度大于2.0mg/L时会使苗芽的质量降低,愈伤组织产生严重,苗芽细小,抽出不明显,颜色发黄甚至变白。附加或不附加100mg/LAdSO4对丛生芽增殖没有明显影响。生根培养基以1/2MS+1.0mg/LIBA+0.5～1.0mg/LIAA为最优,生根率可达100%;在细沙∶土∶椰糠(1∶1∶1)的基质中常规炼苗,成活率可达98%以上。液体纸桥法对胡椒种胚进行培养,在不附加任何生长调节物质的培养基(MS、B5或SH)上只产生单苗,而在附加不同种类和不同浓度的生长调节物质的培养基上则诱导形成愈伤组织,但未能实现分化;以胡椒无菌萌发的实生苗胚轴和叶片切段作外植体进行培养,较易诱导产生愈伤组织,但难以实现分化。     

The isolation of genomic DNA from blackcurrant (ribes nigrum L.)

A method is described for isolating DNA of high molecular mass (M_r) from blackcurrant and other soft-fruit species. Following a hexacethylytimethyl ammonium bromide (CTAB)-based extraction procedure, samples are treated with a glycosidic hydrolase mixture and RNase, and then purified. The suitability of this DNA for Southern analysis and genomic-library construction is demonstrated.     

Plant growth promoting rhizhobacteria (PGPR)-mediated root proliferation in black pepper (Piper nigrum L.) as evidenced through GS Root software

Abstract

Pseudomonas fluorescens strains which are proven biocontrol agents in black pepper against foot rot (caused by Phytophthora capsici ) were also found to enhance root proliferation and fibre root production. Experiments conducted in the greenhouse with five efficient strains of P. fluorescens (IISR-6, IISR-8, IISR-11, IISR-13 and IISR-51) showed that the bacterial strains could significantly increase the root biomass of the plants (30 – 135%). Parameters for total root length, root area and root tips were estimated by scanning the entire root system and analysis through GS Root® software (PP systems, Winterstreet, USA). All the strains increased the root length in the treated plants (12 – 127%), the highest being with IISR-6, which was on a par with IISR-11 and IISR-51. A similar trend was observed with the total root area after bacterization (43 – 200%). The P. fluorescens treated plants had a higher number of feeder roots as evidenced by the increased number of root tips (82 – 137%). The enhanced growth parameters upon root bacterization could be corroborated with the production of the plant growth hormones IAA & GA by the bacterial strains and their P-solubilization potential.     

Antioxidant efficacy of black pepper (Piper nigrum L.) and piperine in rats with high fat diet induced oxidative stress

The present study was aimed to explore the effect of black pepper (Piper nigrum L.) on tissue lipid peroxidation, enzymic and non-enzymic antioxidants in rats fed a high-fat diet. Thirty male Wistar rats (95-115 g) were divided into 5 groups. They were fed standard pellet diet, high-fat diet (20% coconut oil, 2% cholesterol and 0.125% bile salts), high-fat diet plus black pepper (0.25 g or 0.5 g/kg body weight), high-fat diet plus piperine (0.02 g/kg body weight) for a period of 10 weeks. Significantly elevated levels of thiobarbituric acid reactive substances (TBARS), conjugated dienes (CD) and significantly lowered activities of superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and reduced glutathione (GSH) in the liver, heart, kidney, intestine and aorta were observed in rats fed the high fat diet as compared to the control rats. Simultaneous supplementation with black pepper or piperine lowered TBARS and CD levels and maintained SOD, CAT, GPx, GST, and GSH levels to near those of control rats. The data indicate that supplementation with black pepper or the active principle of black pepper, piperine, can reduce high-fat diet induced oxidative stress to the cells.     

水稻线粒体丝氨酸羟甲基转移酶基因的电子克隆

采用基于EST的电子克隆方法得到了一段长1611bp的cDNA序列,以此为信息探针搜索HTGs数据库,找到一个与之高度匹配的基因组DNA序列——OSJNBa0057G07克隆。用FGENESH分析该克隆中的联配区域得到一个包含14个外显子和13个内含子的基因。该基因位于水稻第3染色体物理图谱的136．0～137．6cM区域。推导的ORF编码498个氨基酸,经BLASTP搜索SWISS-PROT数据库和蛋白序列的亚细胞定位显示,该基因编码水稻的线粒体丝氨酸羟甲基转移酶(mSHMT)。该基因受到EST序列的完全支持,其中不乏来自盐胁迫、稻瘟病菌侵染等逆境处理的EST序列,推测该基因与水稻对逆境的应答反应有关。     

Characterisation of novel S-alleles from cherry (Prunus avium L.)

In plant populations exhibiting gametophytic self-incompatibility, individuals harbouring rare S alleles are likely to have a reproductive advantage over individuals having more common alleles. Consequently, determination of the self-incompatibility haplotype of individuals is essential for genetic studies and the development of informed management strategies. This study characterises six new S alleles identified in wild cherry (Prunus avium L.). Investigations to determine the S genotype of individuals in recently planted woodland through length polymorphisms of introns associated with the stylar S-RNase gene and the pollen SFB gene revealed six S intron profiles which did not correspond to those of known S alleles. These are now attributed to S ₂₇ to S ₃₂ . Consensus primers, annealing in the S-RNase sequence coding for the signal peptide and C5 regions, were used to isolate the S-RNase alleles associated with the novel S intron profiles. The proteins corresponding to the new alleles were separated by isoelectric focusing from stylar extracts and their pI values determined. Similarities between the deduced amino acid sequence for the new alleles isolated and other cherry S-RNase sequences available on the databases ranged from 40% to 86%. Amplification products for SFB introns ranged from 172 to 208bp. New sequence regions exposed to positive selection were identified and the significance of the PS3 region reinforced. A phylogenetic relationship between P. avium S-RNases for S ₁₀ and S ₁₃ and between corresponding SFB alleles may indicate co-evolution of allele specificities of these two genes. The nucleotide sequences reported in this paper have been submitted to the EMBL/GenBank database under the following accession numbers: S ₂₇ (DQ266439), S ₂₈ (DQ266440), S ₂₉ (DQ266441), S ₃₀ (DQ266442), S ₃₁ (DQ266443), S ₃₂ (DQ266444).     

辣椒株高遗传分析

以辣椒矮秆自交系B9431（P1）和高秆自交系‘吉林长椒’（P2）为双亲,构建P1、F1、P1、B1、B2和F2 6个家系世代群体,应用植物数量性状主基因＋多基因混合遗传模型对该6个世代群体株高进行多世代联合分析,结果显示：株高遗传符合1对主基因＋多基因遗传模型,高秆对矮秆表现为不完全显性,F1代株高的势能比值为0．39,显性程度为0．91。B1、B2和F2群体主基因遗传率分别为20．35％、17．20％和35．29％,多基因遗传率分别为5．08％、19．75％和0;主基因效应表现为负向加性效应,其值为-6．43,显性效应为0;多基因加性效应值和显性效应值分别为-8．89和9．77。研究还表明,主基因与多基因间的基因效应存在一定差异,主基因加性效应值相当于多基因加性效应值的72．33％,主基因无显性效应,显性效应是由多基因控制遗传。