一条卡瓦胡椒特异RAPD 带转化成SCAR 标记的研究

采用27 份不同来源的胡椒属( Piper) 材料和1 份不同属的草胡椒( Peperomia pellucida) 材料用引物OPQ-03 扩增得到一条约400 碱基对( bp) 卡瓦胡椒特异片段。对该片段进行了克隆和序列分析, 并根据序列分析结果将上述RAPD 分子标记转化为重复性和特异性更好的SCAR ( sequence characterized amplified regions, 序列特征化扩增区) 分子标记。本研究设计出了1 对卡瓦胡椒特异SCAR 引物P7. 1 ( 5′-GGT CAC CTC ACC GCA GCA GGA TGA ACG-3′) 和P7 . 2 (5′-GGT CAC CTC AAT GAC ATG GGA TGA ATC-3′) , 用这对特异引物对本次试验的28 份材料进行PCR 扩增, 结果只有不同属的草胡椒材料无任何扩增, 其它材料均扩增出了预期大小440 bp 的特异带。     

一条卡瓦胡椒特异RAPD带转化成SCAR标记的研究

采用27份不同来源的胡椒属(Piper)材料和1份不同属的草胡椒(Peperomia pellucida)材料用引物OPQ-03扩增得到一条约400碱基对(bp)卡瓦胡椒特异片段。对该片段进行了克隆和序列分析,并根据序列分析结果将上述RAPD分子标记转化为重复性和特异性更好的SCAR(sequence characterized amplifiedre-gions,序列特征化扩增区)分子标记。本研究设计出了1对卡瓦胡椒特异SCAR引物P7.1(5′-GGT CAC CTC ACC GCA GCA GGA TGA ACG-3′)和P7.2(5′-GGT CAC CTC AAT GAC ATG GGA TGA ATC-3′),用这对特异引物对本次试验的28份材料进行PCR扩增,结果只有不同属的草胡椒材料无任何扩增,其它材料均扩增出了预期大小440bp的特异带。     

卡瓦胡椒SCAR标记的研究

目的：采用6份卡瓦胡椒材料、21份栽培胡椒和野生胡椒材料、1份不同属的草胡椒材料共计28份试验材料,开发1对特异SCAR引物。方法：在对它们进行了RAPD研究的基础上,通过克隆、测序和引物设计进行了SCAR分子标记研究。结果：本研究开发了1对特异SCAR引物P10．1和P10．2,用这对特异引物对本次试验的28份材料进行PCR扩增,结果显示,6份卡瓦胡椒材料扩增出了三条带,三条带离的较近,中间一条为预期494bp特异片段。其它胡椒属材料均扩增出一条494bp的特异带,而不同属的草胡椒无任何扩增。结论：这说明引物P10．1和P10．2适用于卡瓦胡椒的分子鉴定(三条带),也可用于胡椒属植物的分子鉴定(一条带),这对卡瓦胡椒种质资源的真伪鉴定及胡椒属植物的分子分类有一定帮助。     

卡瓦胡椒SCAR分子标记的研究

目的:采用6份卡瓦胡椒材料、21份栽培胡椒和野生胡椒材料1、份不同属的草胡椒材料共计28份试验材料,开发1对特异SCAR引物。方法:在对它们进行了RAPD研究的基础上,通过克隆、测序和引物设计进行了SCAR分子标记研究。结果:研究开发了1对卡瓦胡椒特异SCAR引物P4.1和P4.2,用这对特异引物对试验的28份材料进行PCR扩增,结果只有6份卡瓦胡椒材料扩增出了预期大小562bp的特异带,其它材料均无任何扩增。结论:这说明引物P4.1和P4.2为卡瓦胡椒特异SCAR引物,可用于卡瓦胡椒的分子鉴定,这对卡瓦胡椒种质资源的真伪鉴定有一定帮助。     

卡瓦胡椒及胡椒的RAPD聚类分析

目的：通过RAPD分析搞清楚卡瓦胡椒与胡椒及胡椒属其它近缘野生种的亲缘关系。方法和结果：采用随机引物80条进行筛选,用从80条随机引物中入选的20条引物,对卡瓦胡椒和胡椒属共计28份材料进行RAPD扩增,均能产生清晰的扩增谱带。重复1—2次,结果稳定可靠。20个引物共扩增出170个条带,其中多态性条带有20条,占总扩增条带数的12％。RAPD分析结果显示在相似系数0．36处对28份种质可划分为6个类型,其中卡瓦胡椒被单独聚为一类,说明卡瓦胡椒与胡椒及其它近缘野生种的亲缘关系有一定的距离。     

Floristic Geography of Piper (Piperaceae) in China

Piper is the largest genus and important Pantropical components of the Piperaceae family with approximately 1000-2000 species in the world. China is in the northern verge of its natural distribution. In this study, floristic geography of the Piper genus in China was analyzed to reveal its botanical characteristic, origins of speciation and its dispersal channels. The study results showed Yunnan province is one of the very important distribution regions for Piper species, served as the center of the Piper species dispersal toward to the eastern and northern regions of China with gradually reduced species numbers. The similarity level appeared to be low for its species among different provinces, and the Piper distribution regions could be approximately clustered into three clades: tropical clade, subtropical clade and Taiwan clade. Cladistically, Piper species found in China have the highest proportions which were endemic to China, and then followed by Piper species distributed in Tropical Asia, only few species belonged to Pantropical and East Asia areal types. The floristic geography of Piper genus in China is related to that of Indo Chinese Peninsula, Java Islands and Indian Peninsula in some degrees, but not closely related. This study revealed Yunnan province is one of the central of origins for the Piper species, and or one of the central of regions for its species diversification, and the Piper species of Taiwan mainly came from Philippine islands and further diversified into Taiwan clade. Our study results presented a useful method for the systematic studying of Piper genus taxonomy and the evolution of Piper speciation.     

卡瓦胡椒RAPD反应体系的建立

卡瓦胡椒RAPD分子标记的研究,目前国内外尚未见有报道。该试验通过CTAB法提取卡瓦胡椒基因组DNA,通过对模板DNA用量、Mg^2＋浓度、退火温度、电泳上样量等几个单因子试验来建立RAPD稳定扩增体系和反应条件,RAPD扩增结果重复性好,稳定可靠,为卡瓦胡椒RAPD分子标记的研究打下基础。     

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.     

Areas of endemism and distribution patterns for Neotropical Piper species (Piperaceae)

Aim The study aimed to establish areas of endemism and distribution patterns for Neotropical species of the genus Piper in the Neotropical and Andean regions by means of parsimony analysis of endemicity (PAE) and track‐compatibility analysis. Location The study area includes the Neotropical region and the Northern Andean region (Páramo‐Punan subregion). Methods We used distribution information from herbarium specimens and recent monographic revisions for 1152 species of Piper from the Neotropics. First, a PAE was attempted in order to delimit the areas of endemism. Second, we performed a track‐compatibility analysis to establish distribution patterns for Neotropical species of Piper. Terminology for grouping Piper is based on recent phylogenetic analyses. Results The PAE yielded 104 small endemic areas for the genus Piper, 80 of which are in the Caribbean, Amazonian and Paranensis subregions of the Neotropical region, and 24 in the Páramo‐Punan subregion of the Andean region. Track‐compatibility analysis revealed 26 generalized tracks, one in the Páramo‐Punan subregion (Andean region), 19 in the Neotropical region, and six connecting the Andean and Neotropical regions. Both the generalized tracks and endemic areas indicate that distribution of Piper species is restricted to forest areas in the Andes, Amazonia, Chocó, Central America, the Guayana Shield and the Brazilian Atlantic coast. Main conclusions Piper should not be considered an Andean‐centred group as it represents two large species components with distributions centred in the Amazonian and Andean regions. Furthermore, areas of greater species richness and/or endemism are restricted to lowland habitats belonging to the Neotropical region. The distribution patterns of Neotropical species of Piper could be explained by recent events in the Neotropical region, as is the case for the track connecting Chocó and Central America, where most of the species rich groups of the genus are found. Two kinds of event could explain the biogeography of a large part of the Piper taxa with Andean–Amazonian distribution: pre‐Andean and post‐Andean events.     

芽孢杆菌TS02特异RAPD标记的SCAR标记转化和验证

利用自主分离的蜡状芽孢杆菌菌株TS02, 采用RAPD方法对TS02及其同源性相近的5株芽孢杆菌(地衣芽孢杆菌、枯草芽孢杆菌、凝结芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌) 进行了RAPD条带特异性分析, 从TS02基因组中筛选获得了一个533 bp的特异RAPD标记TSR1。TSR1克隆、测序后, 根据其序列设计出一对特异引物P1/P2进行扩增, 结果只在TS02中扩增得到目的片段, 而其余对照菌株扩增为阴性, 从而证明试验得到了在种水平上对该菌种进行准确鉴定的特异SCAR标记。     

黑麦基因组特异DNA片段的分离与SCAR标记的建立

以2个栽培黑麦、2个野生黑麦和4个普通小麦为材料,从200条10碱基RAPD随机引物中筛选出1条引物H11。H11在小麦中有1条低拷贝扩增,而在黑麦中却有极高拷贝的扩增。对H11在黑麦中的高拷贝片段进行克隆、测序,得其全长679 bp,记作OPH11679。根据OPH11679设计特异PCR引物H11-F和H11-R,对小麦族其它物种和含黑麦染色质的物种进行验证,结果发现仅含黑麦染色质的物种能扩增出长为643 bp的片段(命名为pScH643),这表明该片段为黑麦所特有。用H11-F和H11-R对1套中国春-Imperial黑麦附加系等进行扩增,结果显示pScH643片段分布在黑麦整套染色体上,这一结果在小麦-黑麦异源材料的分析中得到进一步验证。即表明pScH643片段可作为SCAR标记用于含黑麦染色质材料的检测。     

芽孢杆菌TS02特异RAPD标记的SCAR标记转化和验证

利用自主分离的蜡状芽孢杆菌菌株TS02,采用RAPD 方法对TS02及其同源性相近的5株芽孢杆菌(地衣芽孢杆菌、枯草芽孢杆菌、凝结芽孢杆菌、巨大芽孢杆菌、短小芽孢杆菌)进行了RAPD条带特异性分析,从TS02基因组中筛选获得了一个533 bp的特异RAPD标记TSR1.TSR1克隆、测序后,根据其序列设计出一对特异引物P1/P2进行扩增,结果只在TS02中扩增得到目的片段,而其余对照菌株扩增为阴性,从而证明试验得到了在种水平上对该菌种进行准确鉴定的特异SCAR标记.     

胡椒、芒果和黄皮的化感作用潜力

以胡椒、芒果和黄皮作为供体,玉米、黄豆、南瓜、花生、萝卜、稗草、马唐、柱花草为受体,通过种子萌发法和根生长法测定胡椒、芒果和黄皮的化感作用潜力.结果表明,玉米、黄豆、南瓜、稗草和马唐的萌发和根生长都受到3种供体水浸提液的影响,3种供体水浸提液在高浓度下抑制而在低浓度下则促进受体的萌发和根生长;胡椒和芒果根系周围的土壤对玉米萌发和根生长有促进作用而对花生的萌发和根生长则有抑制作用.3种供体的水浸提液通过乙酸乙酯、正丁醇萃取分离成不同极性的三相后分别对玉米、萝卜和柱花草进行处理.结果表明,胡椒和芒果浸提液的水相与正丁醇相对受体的抑制作用明显强于乙酸乙酯相,表明胡椒和芒果的化感物质具有较强的极性.     

Taxonomic notes on the genus Piper (Piperaceae)

Sixteen lectotypifications of Asian Piper species are provided. Piper argyrites, P. baccatum, P. leptostachyum, P. majusculum, P. peepuloides, P. quinqueangulatum and P. sulcatum are accepted as species and many new synonyms are proposed. Useful diagnostic characters are described and geographical distribution data of each species are provided.     

Stem diversity, cauline domatia, and the evolution of ant-plant associations in Piper sect. Macrostachys (Piperaceae)

Plants possess a variety of structures that harbor ant nests, and the morphology of these domatia determines the nature of ant-plant mutualisms in a given plant species. In this study, we report on the differences in anatomy between myrmecophytes of Piper, which are regularly excavated by an obligate ant mutualist (Pheidole bicornis) and nonmyrmecophytes of Piper, which consistently have solid stems. Stems of excavated plant species lacked outward evidence of modification; however, striking anatomical differences were apparent between hollow-stemmed species before excavation and the remainder of the solid-stemmed species studied. Prior to excavation by ants, stems of myrmecophytes were characterized by strongly heterogeneous piths in which a large, central area had relatively large cells lacking intracellular crystals with a periphery of smaller cells containing numerous crystals. The domatium excavated by the ants was restricted to the large-celled region. This is the first report of the absence of crystals in ant-excavated portions of stems of myrmecophytes. Cauline domatia became lined with 3-8 cell layers of suberized wound tissue, which may have an impact on nutrient absorption by Piper myrmecophytes.     

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。     

Piper protrusum (Piperaceae), a new species from southern Thailand based on morphological and molecular evidence

Abstract Piper protrusum Chaveer. & Tanee, sp. nov. is described and illustrated. It dominantly comprises three branching types with three different types of leaf blades, bases, and apexes. The critical distinguishing character is the protruded receptacle having a bract and nine stamens. Individual plants have been discovered in areas of Southern Thailand since 2004 without reproductive parts. The investigated sites were revisited several times, and an individual with flowers was finally found in July 2009. Phylogenetic analysis of the new species and five similar species is carried out based on DNA fingerprinting. The genetic distances between the new species and five similar species range from 0.25 to 0.35, supporting new species designation. Molecular data conform to morphological data in validating that it is a new species. Additionally, its DNA barcodes have been provided for further identification in case the morphological data is unclear. The sequence data have been submitted to the GenBank database under accession numbers GU980898 (rpoB gene), GU980899 (rpoC1 gene), and GU980900 (psbA‐trnH region).     

Patterns of genotypic variation and phenotypic plasticity of light response in two tropical Piper (Piperaceae) species

Patterns of phenotypic plasticity and genotypic variation in light response of growth and photosynthesis were examined in two species of rain forest shrub that differ in ecological distribution within the forest. We further examined correlations among photosynthetic and growth traits. We hypothesized that the pioneer species, Piper sancti-felicis, would display greater phenotypic plasticity than the shade-tolerant species, Piper arieianum. We further proposed that, in both species, genotypic effects would be more apparent in growth-related traits than photosynthetic traits due to more concentrated selection pressure on gas-exchange traits. P. sancti-felicis did not demonstrate greater phenotypic plasticity of light response. Although many of the traits measured had significant genotype effects, neither species showed any significant effects of genotype on light response of photosynthesis, suggesting little genetic variation for this trait within populations. A principal components analysis clearly illustrated both species and light effects, with the treatments dividing neatly along the axis of the first principal component and the species separating along the second principal component axis. Results indicated general similarities between the species in their trait correlation structure and level of integration among traits, but characteristic differences were observed in the patterns of change between low and high light. Both species had more correlations than expected within groups of growth-related or photosynthetic traits; strong correlations of traits between these two groups were underrepresented. The similar pattern of genetic variation and phenotypic integration observed in these two congeners may be due more to their close phylogenetic relation than to their ecological distributions.