Molecular Identification and Cultivar Fingerprints of Prunus persica （L.） Batsch Germplasms

[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica （L.） Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites （simple sequence repeats,SSRs） and Inter-Simple Sequence Repeat （ISSR） markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region （SCAR） markers.The genetic similarity coefficient （GS） estimated from these molecular data averaged were 0.939 （ranged from 0.856 to 0.983） for ISSR and 0.646 （ranged from 0.240 to 1.000） for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.     

Construction and Characterization of a cDNA Library from the Pulp of Coconut （Cocos nucifera L.）

To investigate the gene expression profile of endosperm development, a cDNA library was constructed and characterized from the pulp of coconut at different developmental stages. The constructed cDNA library incorporated approximately 1 × 10^7 clones in total, and the size of the insertion fragments ranged from 800 to 2 000 bp. Sequencing results of 100 randomly picked clones showed that the recombination rate was 96%. In subsequent sequence analysis, 41 clones （41%） were homologous to known function proteins, and 23 clones showed high amino acid identity （more than 80%） with the corresponding genes of different plants. Semi-quantitative RT-PCR indicated that oleosin and globulin genes are pulpspecific expression, and have differential expression level in different developmental stage. Clone 29, recognized as homologous to KIAA1239 protein （Homo sapiens）, was observed to occur nine times, indicating that this gene may be over-expressed during the endosperm development stage. However, the homologous protein was found only in mammals, and the detailed function is still unknown. Elucidation of the functional characterization of these genes will be carried out immediately.     

Identification of Organic Substances Associated with Tissue Senescence in Upland Cotton （Gossypium spp. L.） Based on GC-MS Analysis

Premature senescence in crop production,especially occurred at the late growth stage,generally results in a reduction in yield and quality.Therefore,it is beneficial for yield and quality to properly delay senescence of plant tissues during the late developmental stage.In this study,it was observed that the chlorophyll content and photosynthetic rate were gradually decreased along leaf growth progression,and the rates of reduction were promoted by drought.Based on gas chromatography-mass spectrometry (GC-MS) analysis,total eight,five,seven,and five kinds of organic compounds that putatively associated with the tissue senescent progression were identified in leaves,fruit branches,petals,and sepals,respectively.It was found that the identified organic compound,such as α-pinene,β-pinene,and pentadecane were present in different tissues.Among the total ten organic substances identified to be related with the leaf senescence,half were specifically detected in the drought treatment.These results suggest some biochemical pathways associated with the leaf senescence are distinctly regulated by drought.The identified organic compounds in the tested tissues showed three types on the performance pattern based on the contents along with the senescent progression,including gradually increasing,decreasing,and a curve with one single peak.Thus,during the senescence process in tissues,a subset of metabolic substances occur modifications on the quantities,reflecting a complicate biochemical reactions are initiated via the senescence signals.Further analysis of the important organic substances will be helpful for elucidation of the tissue senescence mechanism at the biochemical level and provide a new insight of the senescence signaling transductions in cotton.     

Isolation and Expression Analysis of Two Genes Encoding Cinnamate 4-Hydroxylase from Cotton （Gossypium hirsutum）

Two genes （GhC4H1 and GhC4H2） that encode putative cotton cinnamate 4-hydroxylases that catalyze the second step in the phenylpropanoid pathway were isolated from developing cotton fibers. GhC4H1 and GhC4H2 each contain open reading frames of 1 518 base pairs （bp） in length and both encode proteins consisting of 505 amino acid residues. They are 90.89% identical to each other at the amino acid sequence level and belong to class I of plant C4Hs. GhC4H1 and GhC4H2 genomic DNA are 2 247 and 2 161 bp long, respectively, and contain two introns located at conserved positions relative to the coding sequence. GhC4HI and GhC4H2 promoters were isolated and found to contain many cis-elements （boxes P, L and AC-1 element） previously identified in the promoters of other phenylpropanoid pathway genes. Histochemical staining showed GUS expression driven by the GhC4H1 and GhC4H2 promoters in ovules and fibers tissues. GhC4H1 and GhC4H2 were also widely expressed in other cotton tissues. GhC4H2 expression reached its highest level during the elongation stage of fiber development, whereas GhC4H1 expression increased during the secondary wall development period in cotton fibers. Our results contribute to a better understanding of the biochemical role of GhC4H1 and GhC4H2 in cotton fiber development.     

Molecular Identification and Expression Analysis of GhHYDRA 1 Gene,a Homologue of HYDRA1 Gene From Upland Cotton （Gossypium hirsutum L.）

Beside as precursors of BRs biosynthesis,more and more evidences supposed that phytosterols play an important role in plant growth and development.To investigate the effects of phytosterols on the fiber development of upland cotton(Gossypium hirsutum L.)and the molecular base of sterol regulating cotton fiber growth,a homologue of HYDRA1 was cloned from upland cotton(cv.Xuzhou 142)by screening cotton fiber EST database and contigging the candidate ESTs.The GhHYDRA1 encoded a polypeptide of 218 amino acid residues and the deduced amino acid sequences had high homology with the members of HYDRA1 in Populus trichocarpa,Solanum tuberosum,and Arabidopsis thaliana.Moreover,GhHYDRA1 had comparable transmembrane regions to AtHYDRA1 in sequence,length,order,and spacing,except for a C-terminal polylysine cluster.Quantitative real-time RT-PCR analysis revealed that the higher expression levels of GhHYDRA1 gene were detected in 6 to 12 DPA(days post anthesis)fibers,while the lower levels were observed in 0 DPA ovule(with fibers)and 16 to 18 DPA fibers.These results indicated that GhHYDRA1 is the homologue of HYDRA1 gene and plays a crucial role in fiber elongation.Furthermore,auxin and BL up-regulated the expression level of GhHYDRA1 while ABA and KT down-regulated the expression level of GhHYDRA1 in cotton ovule and fiber growth.The result suggested that phytosterols play a role in the interaction of plant hormones.     

Resistance to the Whitefly Aleurotrachelus socialis（Hemiptera：Aleyrodidae） and SSR Marker Identification in Advanced Populations of the Hybrid Manihot esculenta subsp.Manihot flabellifolia

Genes resistant to Aleurotrachelus socialis were transferred to the F 1 from the interspecific hybrid wild species of Manihot flabellifolia to M.esculenta and two advanced generations of backcrosses（BC 1 and BC 2）.We characterized the resistance of A.socialis transferred to BC 2 parents（CW67-160,CW67-130,CW67-44）,MTAI-8（BC 1）,resistant（CMB9B-73） and susceptible（CMB9B-104） genotypes from contrasting pools,and resistant（MEcu-72） and susceptible（CMC-40） genotypes.Whitefly demography and biology were evaluated.SSR molecular markers associated with a phenotypic response of plant resistance were detected in segregating populations（BC 2）.Results showed that although female survival time was similar on all hosts,the lowest averages of longevity,fecundity and oviposition rate were observed in the resistant control MEcu- 72,only being significantly similar to the parent CW67-130.When the BC 1 and BC 2 populations were compared,it was found that A.socialis fecundity was eight times lower on CMB9B-73 progeny than on CW67-130,expressing the highest levels of resistance to the whitefly.Ten genotypes of CMB9A and CMB9B family had the best segregation.A total of 486 microsatellite primers were evaluated using bulked segregant analysis（BSA）,11 showed polymorphism between the contrasting pools and only one showed significant differences between resistant and susceptible individuals.In conclusion,fecundity was the parameter that impacted most on the intrinsic rate of A.socialis population growth.     

A Simple Method for the Isolation and Purification of 2,4-Dihydroxy-7-Methoxy-2H-1,4-Benzoxazin-3（4H）-One （DIMBOA） from Maize （Zea mays L.） Seedlings

2,4-Dihydroxy-7-methoxy-2H-1,4-benzoxazin-3(4H)-one (DIMBOA), the dominant benzoxazinoid hydroxamic acid in maize (Zea Mays L.), serves as important factors of resistance against insects and microbial diseases, allelochemicals used in competition with other plants. In this paper, a novel and simple method for the isolation and purification of DIMBOA from maize seedlings was developed. Frozen shoots from 7-d-old maize seedlings (1 000×g) were firstly defrosted and then were directly homogenized and extracted with ethyl acetate. The macerate was allowed to stand at room temperature (25±2)°C for 1 h to allow enzymatic release of DIMBOA from DIMBOA-glucoside. Then the ethyl acetate phase was filtered, dried and evaporated to dryness. The resulting light-tan, semicrystalline residue was stored at -20°C for 24 h. Upon recrystallization from acetone-hexane, a relative higher yield (0.58 g) of pure DIMBOA crystals was obtained compared with the yield afforded by Woodward methodology (0.26 g).     

黄顶菊中除草活性成份Ⅱ作用机制的初步研究

为了控制和利用外来入侵杂草黄顶菊,并发现新的除草作用机制,本试验通过水蒸气蒸馏的方法从黄顶菊花中提取并分离出一种除草活性很强的成份Ⅱ｡在此基础上测定了成份Ⅱ对马唐叶片膜透性､叶绿素含量､ β-胡萝卜素､丙二醛､超氧自由基､含水量､根系活力的变化,以明确其除草作用机制｡结果表明:成份Ⅱ在光照条件下对马唐叶片膜透性､叶绿素含量､β-胡萝卜素､丙二醛､超氧自由基的影响均高于其在黑暗条件下对这些物质的影响｡成份Ⅱ是一种光活化物质,其对杂草的除草活性主要是影响了杂草的光合作用｡本试验为发现新的除草剂作用靶标奠定了基础｡     

黄顶菊化感物质释放途径的初步研究

采用室内生物测定法,以萝卜、小麦、玉米和棉花为受体,研究了黄顶菊对受体植物的化感作用,旨在了解黄顶菊化感物质的释放途径。结果显示:黄顶菊自然挥发物对受体植物不表现化感作用;黄顶菊淋溶物对棉花幼苗生长表现出明显的抑制作用,对棉花幼苗高度和根长的抑制率分别为6.29%和10.42%;黄顶菊枯落物浸提液和根系分泌物浸提液对4种受体植物均表现出不同程度的抑制作用,除了黄顶菊根系分泌物浸提液B3处理外,其它处理的综合效应均显著大于淋溶物处理,如黄顶菊枯落物浸提液(A1、A2、A3)、根系分泌物浸提液(B1、B2、B3)以及淋溶物等各处理对受体植物的综合效应分别为44.74、29.37、14.5、31.54、16.52、3.55、1.48。结果表明,黄顶菊主要通过植株残体和根系分泌向环境中释放化感物质,其次是通过雨雾的淋溶对受体植物产生化感作用。     

外来入侵植物黄顶菊的化感作用初步研究

为明确黄顶菊对其他植物的化感作用,利用黄顶菊茎叶的不同提取物对几种植物的生长影响进行了测定。结果表明,黄顶菊茎叶水提物对玉米、小麦、棉花、大豆、花生、马唐和反枝苋都表现出了不同程度的抑制作用,其中以对棉花的化感效应最强,在其浓度为0.2 g/mL时,对根长和茎长的化感指数分别为-0.85和-0.88,而该水提物对水稻的生长则表现为促进作用。黄顶菊茎叶的石油醚、氯仿、乙酸乙酯、丙酮和乙醇的提取物,对几种植物的化感效应要高于水提物,其中以丙酮提取物的化感效应最强。利用重结晶法从黄顶菊茎叶的丙酮提取物中得到了一个具有除草活性的物质,该物质的熔点为192.5℃~193.5℃,最大吸收波长为220 nm,初步判断该化合物是一种碱性有机物,该物质在浓度为1000,500,100,50 mg/L时,对马唐,反枝苋生长均有抑制作用。     

黄顶菊对几种作物生长影响的作用机理初步研究

在室内土壤盆栽条件下, 探讨了黄顶菊对附近生长的萝卜、玉米、黄瓜和小麦植株生长的影响, 结果发现, 黄顶菊对供试4 种作物种子的萌发以及株高和鲜重增长都有明显的抑制作用, 其中对黄瓜、萝卜的抑制作用更 强。研究还发现, 生长在黄顶菊附近的萝卜、玉米、黄瓜和小麦根系活力明显低于对照, 叶片的可溶性蛋白、叶绿 素含量以及SOD 、POD、CAT 活力均低于对照, 而丙二醛含量高于对照。结果预示黄顶菊可能通过根系分泌物 对附近的作物施加影响。     

黄顶菊种子萌发特性研究

从光照、温度、土壤含水量和空气相对湿度等方面,研究了其对黄顶菊种子萌发的影响。结果表明：光照促进黄顶菊种子萌发;适于黄顶菊种子萌发的温度为25～40℃;土壤含水量在30％～90％时,有利于黄顶菊种子萌发;空气相对湿度对黄顶菊种子萌发影响较小。     

外来入侵植物黄顶菊花中叶黄素的提取研究

为明确黄顶菊花中叶黄素的含量及最佳提取方法,经过溶剂筛选,采用石油醚∶无水乙醇=5∶1为提取溶剂,分别在震荡提取､超声波提取和微波提取3种条件下得到叶黄素酯粗品,然后经皂化处理得到叶黄素粗品｡用高效液相色谱法进行检测以确定黄顶菊花中叶黄素的含量｡结果表明:微波提取效率最高,用高效液相色谱法检测显示叶黄素粗品中叶黄素的含量为83.16%,从而计算出黄顶菊花中叶黄素的含量为8.291mg/100g｡该研究为防控外来入侵植物黄顶菊提供了新的方法,同时丰富了我国叶黄素资源,降低了叶黄素的生产成本｡     

The Extraction,Isolation and Identification of Exudates from the Roots of Flaveria bidentis

Large amounts ofFlaveria bidentis＇s root culturing solution were obtained by using DFT （deep flow technique） equipment and these solution which was vacuum concentrated （10, 20 mg mL-~） can have a certain inhibition on Triticum aestivum, Cucumis sativus, Raphanus sativus, Amaranthus retroflexus, Setaria viridis, Chenopodium album, Echinochloa crusgalli and Chloris virgata. This outcome suggested some active compounds in the root exudates ofFIaveria bidentis can inhibit the germination, seedling elongation and root length. The dichloromethane extract of root exudates was identificated by GC-MS, and 29 kinds of compounds, including esters, hydrocarbons, ketones, thiazole, amines, etc. were obtained and the phthalate n-octyl ester, phthalate 2-ethylhexyl ester were proved to be allelochemicals. The culturing solution of root exudates was separated through the resin column and silica gel column and a component inhibiting seedling height, root length and fresh weight of wheat was got. There were 6 kinds of organic compounds in this component including dioctyl phthalate, 1,2-phthalate, mono（2-ethylhexyl） ester by GC-MS.     

黄顶菊的开花和成熟特性研究

通过人工播种,研究了黄顶菊的开花和成熟特性。结果表明:出苗期早的黄顶菊开花期较早,从出苗到开花需120~50 d,7月下旬开始开花,花期持续到10月下旬;8月之前出苗的黄顶菊均能产生种子,出苗越早,黄顶菊的单株生物量越高,单株生物量与单株种子数量间的相关系数为0.990 7。