Scientia Agricultura Sinica

Microarray Analysis of Gene Expression Profile Related to Grain Storage Metabolism in Rice Endosperms as Affected by High Temperature at Filling Stage

WEI Ke-su,CHENG Fang-min,DONG Hai-tao,ZHANG Qi-fang,LIU Kui-gang,CAO Zhen-zhen

Scientia Agricultura Sinica. 2010, 43(1): 1-11 . doi:10.3864/j.issn.0578-1752.2010.01.001

Abstract ( 1089 )

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【Objective】 Effects of high temperature after rice flowering stage on the expression profile and their patterns of mass genes related to grain storage metabolism were clarified. 【Method】 A cDNA array of 3574 unigenes containing was used for the detection of different filling grain samples and the controlled temperature treatments were conducted in growth chambers, with the daily mean temperature being controlled at 32℃(36℃/28℃) and 22℃(26℃/18℃) for high and moderate temperatures, respectively. 【Result】 The expression levels of majority genes were not sensitive to high temperature treatment among 657-673 candidate genes, with their R value (the ratio of gene hybridized signal between two temperature treatments) ranging from 0.8 to 1.2. However, the different expression genes (up-regulation or down-regulation) increased remarkably when rice plants were consistently exposed to high temperature treatment. There was a considerable difference in gene expression patterns of grain starch synthesis and carbohydrate metabolism induced by high temperature treatment after flowering stage, varied greatly with different gene function classification, metabolizing pathway, iso-form types (or subunit) and also filling stage, in which such genes imposing sugar signal function, with much wide variation in expression levels between two temperature treatment, were more sensitive to high temperature than those genes in relation with grain starch synthesis and other carbohydrate metabolism. The expressing response of storage protein genes to high temperature was mostly down-regulation, despite different patterns for some protein subunit. For all genes related to grain protein synthesis metabolism, these genes impacting ribosomal protein and defense protein synthesis were mostly sensitive to different temperature treatments or strongly-high specific expression induced by high temperature at filling stage. 【Conclusion】 There is a complex regulating network existing in the effects of high temperature stress on mass genes related to grain storage metabolism, with many genes being probably regulated by high temperature.

Analysis of Grain Quality and Superior Quality Rate of Glutinous Rice Cultivars Bred Since the 1980s of the 20th Century in China

MIN Jie,TANG Sheng-xiang,SHI Jian-hua,ZHU Zhi-wei

Scientia Agricultura Sinica. 2010, 43(1): 12-19 . doi:10.3864/j.issn.0578-1752.2010.01.002

Abstract ( 936 )

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【Objective】 To understand the recent status of the grain quality of glutinous rice cultivars and to clarify the key objective for genetic improvement of glutinous rice in China. 【Method】 Seven grain quality traits and superior quality rates (SQR) of 570 glutinous rice cultivars bred since 1980s of the 20th century were analyzed. 【Result】 The results showed that each average values of all seven quality traits had reached or surpassed the standards of the Third Superior Quality of Rice. The SQRs of 7 grain quality traits tested were from 64.4% to 97.9%, with the average of 81.9%. The excellent glutinous rice varieties with all of 7 grain quality traits in or above the standards of the Third Superior Quality,occupied about 45.1% of total 570 tested materials. The average SQRs of the glutinous rice cultivars bred in 1980s and 1990s of the 20th century and 2001-2007 were of 81.5%, 81.0%, and 84.3%, respectively. Compared with the 1980s, the glutinous rice cultivars bred in 2001-2007 had higher SQR in brown rice rate (BRR), gelatinization temperature (GT) and amylose content (AC), higher by 6%, 11.8%, and 13.4%, respectively, showing the grain quality of the glutinous rice cultivars bred in 2001-2007 was better than those both in 1980s and 1990s. It was noted that the SQRs of head rice rate (HRR) in 1980s, 1990s and 2001-2007 were of 71.0%, 58.3%, and 62.9%, respectively, all lower than that of other 6 rice quality traits. The grain quality of japonica glutinous rice cultivars, in general, was better than that of indica ones. Regarding to the rice growing areas, the varieties of glutinous rice in the Central and North China had better grain quality than that in the South west and South China. 【Conclusion】 It is concluded that the grain quality of glutinous rice cultivars bred in the past recent 25 years in China is rather good. However, the grain quality of glutinous rice should be continuously improved, especially in raising further head rice rate.

Mapping and Analysis of an Oil Content QTL in the Linkage Group 7 Using BC3F1 Population of Brassica napus

SHAO Yu-suo,NI Xi-yuan,HUANG Ji-xiang,REN Li-ping,ZHAO Wei,FANG Xiao-yan,CAO Ming-fu,ZHAO Jian-yi

Scientia Agricultura Sinica. 2010, 43(1): 20-28 . doi:10.3864/j.issn.0578-1752.2010.01.003

Abstract ( 1038 )

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【Objective】 Mapping and conforming a major QTL for oil content in linkage group 7 using BC3F1 population in Brassica napus L. 【Method】 QTL-near isogenic lines (NILs) were developed by marker assistant selection on the basis of primary QTL analysis with SG-DH lines. 1700 BC3F1 plants, containing corresponding marker genotypes on A7 and trait phenotypes of oil content, were used for fine mapping oil QTL and association analysis between the marker genotypes and the trait (oil content) performance on A7 by software WinQTLCartographer 2.5 and SPSS11.5. 【Result】 The confidence interval of oil content QTL was located between the markers ZAAS849s and R131, which range about 21.7 cM and the peak value of LOD was 9.71. The distances between QTL and the nearest flanking markers RPSaA3 and ZAAS839 were 0.9 and 2.1 cM, respectively. The additive effect of QTL was 0.75, with Chinese allele increasing oil content. Marker-oil content association analysis (ANOVA) indicated that the oil content showed a significant difference among the three types of marker genotypes in each loci within QTL region (four markers), of which the marker ZAAS839 showed the largest P value (P=1.2×10-10). 【Conclusion】 QTL position identified by mapping analysis using BC3F1 population was overlapped with primary QTL identification using SG-DH lines, however, the confidence interval was clearly decreased. The most probable QTL position is in the region near the markers RPSaA3 and ZAAS839. The genetic distance between RPSaA3 and ZAAS839 is about 3 cM.

The Ultrastructural Changes of Female Gametophyte During Its Development in the Apomictic Monosomic Addition Line M14 of Beta corolliflora in Sugar Beet

SHANG Ya-jia,SHEN Jia-heng,GUO De-dong,LI Wei,DING Chang-hong,LU Jun-ping

Scientia Agricultura Sinica. 2010, 43(1): 29-38 . doi:10.3864/j.issn.0578-1752.2010.01.004

Abstract ( 963 )

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【Objective】This study explored the ultrastructural changes of female gametophyte during its development in the apomictic monosomic addition line M14 of Beta corolliflora in sugar beet. 【Method】 The ultrastructural changes were investigated by electron microscopy. 【Result】 The monosomic addition line M14 is a facultative apomict. The diplosporous female gametophytes are of Antennaria type and Allium odorum type sexuality is of Polygonum type. Antennaria type is the main one and its ultrastructural characteristics are that the development of female gametophyte was rapid, from the functional megaspore until the cellularized embryo sac, the category and amounts of organelles present the following growth trend, i.e. nucleolus which are more big had nucleolus vacuoles, nuclear pores are obvious; there are many ribosome; also a lot of mitochondria in the development of female gametophyte, many tubes appear from the two-nucleate embryo sac, but transform into electron-transparent in eight-nucleate embryo sac, then recovered as former. The number of plastids was stable and their shapes were varied, some of them contained starch grains. The endoplasmic reticulum had many branches or were interwove as ropes near nucleus, vacuoles and cell walls. Dictyosomes were relatively rare in the development of female gametophyte, but abundant dictyosome which often associated with many small electron-transparent vesicles was found in the persistent synergid. Some lipid drops were always present near the vacuoles and mitochondrias. Late period of cellularized megagametophyte, two synergids successively degenerated in the great majority of embryo sacs, very few had only one degenerated synergid, with another persistent. Compared with Antennaria type embryo sac, the development of Allium odorum type and Polygonum type were slowly, there were no obvious changes of category and amounts of organelles until the period of one-nucleate embryo sac. Whether had degenerated cell and deposition of callose during the development of functional megaspores and one-nucleate embryo sacs were the characters distinguishing the Antennaria-type gametophyte of female and Allium odorum-type.【Conclusion】 There were distinct characteristics during the development of Antennaria-type, Allium odorum type and Polygonum type gametophyte. From the functional megaspore until the cellularized embryo sac, Antennaria type gametophyte present high rate of metabolic activity, its volume augmented increased with category and amount of organelles. It is supposed that Antennaria type gametophyte dominated the afterward growth. Whether had degenerated cell and deposition of calluses before the development of one-nucleate embryo sacs were the characters distinguishing the female gametophyte of Polygonum type, Antennaria type or Allium odorum type. From functional megaspores to one-nucleate embryo sacs, volume and cytoplasmic mature degree of Allium odorum type and Antennaria type embryo sac present slow-moving growth trend. It is concluded that most sexual female gametophyte degenerated in later developmental stages.

Differences in Nitrogen Absorption and Use Efficiency in Rice Genotypes with Different Yield Performance

YIN Chun-yuan,ZHANG Qing,WEI Hai-yan,ZHANG Hong-cheng,DAI Qi-gen,HUO Zhong-yang,XU Ke,MA Qun,HANG Jie,ZHANG Sheng-fei

Scientia Agricultura Sinica. 2010, 43(1): 39-50 . doi:10.3864/j.issn.0578-1752.2010.01.005

Abstract ( 1051 )

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【Objective】 The objective of this study was to identify the differences in N absorption and use efficiency in rice genotypes with different yield performances. 【Method】 A field experiment was conducted with 20 rice genotypes(10 high-yielding and 10 low-yielding performance) selected from 108 rice genotypes including early (125-135 d, growth duration), medium (136-145 d) and late-maturing (146-155 d) medium japonica and early (156-165 d) and medium-maturing (166-175 d) late japonica rice under 225 kg?hm-2 N fertilizer application in 2006 and 2007 on the farm of Yangzhou University, Jiangsu province, China. Yield, N accumulation, N uptake rate, N use efficiency and their relationship were analyzed. 【Result】 The results showed that the mean yields of high-yielding rice genotypes were 31.6%, 31.94%, 39.47%, 26.21%,and 21.82% higher than those of low-yielding rice genotypes, respectively, with the extension of the growth duration. N accumulation and use efficiency increased with the extension of the growth duration, and which of high-yielding genotypes was significantly higher than those of low-yielding genotypes. N accumulation and uptake rate of high-yielding genotypes were higher than those of low-yielding during the growth phases from transplanting to elongation and from elongation to heading, while it showed a reversed trend during the growth phase from heading to maturing for some genotypes. Correlation analysis indicated that there existed significant or highly significant positive correlations between yield and N accumulation, N use efficiency, N uptake rate during the growth phase from transplanting to elongation.【Conclusion】 Compared with the low-yielding rice genotypes, N accumulation in various growing stages, N uptake rate before heading and N use efficiency of high-yielding rice genotypes were higher. It was obvious that high yield and increasing N accumulation and use efficiency could be coordinated development on improving genotype.

Physiological Mechanism of Maize Hybrids in Response to P Deficiency and Differences Among Cultivars

YUAN Shuo,PENG Zheng-ping,SHA Xiao-qing,WANG Yan-qun

Scientia Agricultura Sinica. 2010, 43(1): 51-58 . doi:10.3864/j.issn.0578-1752.2010.01.006

Abstract ( 1222 )

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【Objective】 The objective of this study was to elucidate the physiological mechanisms of the maize hybrids under P deficiency condition, and to provide a scientific evidence for extension and utilization of cultivars with higher yield and efficiency. 【Method】 Three maize hybrids, Liyu 16, Jidan 28, and Kuancheng 10, were used as the testing materials and subjected to two phosphorus levels: sufficiency and deficiency. Effects of P deficiency on plant development, root morphology, activity and physiological characters were investigated among three cultivars at the seedling stage in hydroponic culture. 【Result】 The results showed that under P-deficient condition, the height, dry weight and P content increment in shoot and whole seedling all decreased while the dry weight of roots and ratio of root to shoot increased. Moreover, root length enhanced by 61%, 62%, and 17%, root radius depressed by 19%, 29%, and 4%, and root surface area increased by 31%, 18%, and 13%, respectively, compared with plants with P. Total uptake area and specific surface area of roots were both low, and pH in the nutrient medium respectively reduced by 1.3, 0.8, and 0.7 units, but the three cultivars all had higher uptake kinetics of H2PO4? and stronger abilities to endure P deficiency. The acid phosphatase (APase) activities in root tissue were improved. However, there were still significant differences among the cultivars. 【Conclusion】Compared with Jidan 28 and Kuancheng 10, Liyu 16 had higher dry weight increment, root length and root surface area, root activity and proton exudation, stronger tolerance ability to P deficiency, larger uptake kinetics of H2PO4– and APase activity in root tissue.

Al Stress-Induced Citrate Secretion from Roots in Stylosanthes

ZUO Fang-hua,LING Gui-zhi,TANG Xin-lian,YU Yong-xiong,LI Yao-yan,LI Xiao-feng

Scientia Agricultura Sinica. 2010, 43(1): 59-64 . doi:10.3864/j.issn.0578-1752.2010.01.007

Abstract ( 1163 )

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【Objective】 To reveal the mechanism responsible for Al-resistance in stylo (Stylosanthes), Al-induced secretion of citrate from roots was investigated. 【Method】 The Al-resistance and the effect of Al on the secretion of citrate from roots in stylo (cv. Reyan 2) with comparison to alfalfa (Medicago sativa L., cv Sunshine 3) were studied by hydroponics. 【Result】 There was no dramatic inhibition on the root elongation of stylo exposed to a 10 μmol?L-1 AlCl3 solution, while root elongation was inhibited significantly and the root apices were strongly stained by cyanine R in alfalfa after exposure of roots to 3 or 5 μmol?L-1 AlCl3 for 24 h. Under Al stress, citrate was secreted from roots of stylo. The amount of citrate secretion in stylo was increased with the increasing Al concentration (10, 20, 30 μmol?L-1 AlCl3) and the duration of Al treatment (6, 12, 18, 24 h), but citrate was not secreted from roots in alfalfa. On the other hand, 10 μmol?L-1 anion channel inhibitors including phenylglyoxal, niflumic acid, 4,4’-disothiocyanastilbene- 2,2’-disulfonic acid, and 9-anthracenecarboxylic acid blocked the secretion in stylo. However, neither Al treatment nor the treatment with A-9-C failed to affect the citrate synthase activity of the root apices significantly in stylo. 【Conclusion】 These results suggest that Al-induced secretion of citrate from roots may be a mechanism responsible for Al-resistance and the anion channels involve in the secretion in the stylo.

Effects of Weak Light on Photosynthetic Characteristics of Peanut Seedlings

ZHANG Kun,WAN Yong-shan,LIU Feng-zhen

Scientia Agricultura Sinica. 2010, 43(1): 65-71 . doi:10.3864/j.issn.0578-1752.2010.01.008

Abstract ( 1031 )

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【Objective】 The aim of this study was to investigate the effects of weak light on the photosynthetic characteristics of peanut leaves at the seedling stage and to offer a theoretical basis for the programming of peanut intercropped with wheat. 【Method】 Fenghua 1 was grown in an experiment with four shading treatments (CK, in which the plants were grown under natural light, and 27% shading, 43% shading, 77% shading) and the investigation was carried out at seedling stage using black sunshade net. The chlorophyll content, net photosynthetic rate, photosynthetic curve, fluorescence parameters and photosynthetic enzyme activities were tested. 【Result】 Shading treatment significantly reduced the net photosynthetic rate (Pn), stomatal conductance(Gs), intercellular CO2 concentration(Ci), light compensation point, light saturation point, CO2 compensation concentration, CO2 saturation concentration, carboxylation efficiency, RUBPCase and PEPCase activities along with the shading degree but increased the chlorophyll content, apparent quantum yield, and ФPSⅡ and Fv/Fm. Real-time low light intensity and long-term shading treatment depressed photosynthetic enzyme activities jointly. 【Conclusion】 Shading at seedling stage depressed Pn significantly, which induced by stomatal limitation and non-stomatal factors (such as decline of photosynthetic ability of mesophyll cell, changes of absorbed light allocation of PSⅡ), improved the capacity of utilization of weak light. Shading of 27% had less influence, and therefore can program plant the standards of intercropping system of wheat and peanut.

A Study on the Inheritance of Resistance to Whitefly in Soybean

XU Ran,LI Wei,ZHANG Li-feng,LIN Yan-hui,QI Bo,XING Han

Scientia Agricultura Sinica. 2010, 43(1): 72-78 . doi:10.3864/j.issn.0578-1752.2010.01.009

Abstract ( 892 )

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【Objective】 Whitefly (Bemisia tabaci Gennadius) is a kind of agricultural pests with wide occurrence, severe harm, strong virulence in soybean around the world, so it is an effective way to breed resistant soybean varieties to control whitefly. This study was to reveal the inheritance of resistance to whitefly in soybean, and provide information for soybean breeding. 【Method】 Two crosses, Qihuang 26×Huapidou and Huapidou×Qihuang 26, were made with the highest resistant germplasm Huapidou and the highest susceptible germplasm Qihuang 26. From the crosses, 4 F2 populations were derived in Jinan and Guanxian of Shandong. The means of whitefly per leaflet (MWPL) of all the populations were identified. Genetic analysis was performed under the major gene + polygene mixed inheritance model in these populations. 【Result】 Resistance of soybean to whitefly was controlled by two major genes and plus polygene, with the major genes inheritability of 86.41%, 85.72%, 95.90%, and 95.26%, respectively. 【Conclusion】The inheritance of resistance to whitefly in soybean was controlled by major genes and polygene, and affected by the environment.

HrcJ is Involved in Type-Ⅲ Apparatus Formation of Xanthomonas oryzae pv. oryzicola for Hypersensitive Response in Nonhost Tobacco and Pathogenicity in Rice

ZHAO Wen-xiang,HAN Yang-chun,CUI Yi-ping,ZHAO Mei-qin,LI Yu-rong,ZOU Li-fang,CHEN Gong-you

Scientia Agricultura Sinica. 2010, 43(1): 79-86 .

Abstract ( 1022 )

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【Objective】 Repertoires of pathogenicity effectors in Xanthomonas oryzae pv. oryzicola are injected into plant cells through type-Ⅲ secretion system (T3SS) which is encoded by the hrp (hypersensitive response and pathogenicity) genes, but it is unclear what roles the hrcJ gene plays in pathogen pathogenesis in rice and in T3SS formation. 【Method】 In this report, the hrcJ gene was knocked out by marker exchange method. 【Result】 It was found that the mutant had lost the ability to trigger HR in tobacco and pathogenicity in rice. Protein-protein interactions, revealed by yeast two-hybrid system, demonstrated that the lipoprotein domain at N-terminal of HrcJ interacted with HrcC and the transmembrane domain at C-terminal interacted with HrcV, indicating that HrcJ was a linker protein between the inner and out membranes of the pathogen cell for T3SS formation. Complementation assays in planta showed that either the deletion in the lipoprotein domain or in the transmembrane domain did not restore HR induction in tobacco and pathogenicity in rice to the hrcJ mutant. The reverse transcriptional polymerase chain reaction (RT-PCR) revealed that the hrcJ gene expression was regulated by the hrpX gene and the expression of the effector gene hpa1 was not affected in the hrcJ mutant. 【Conclusion】 These results suggest that the hrcJ gene is required not only for HR induction in tobacco and pathogenicity in rice, but also in T3SS formation through which pathogenicity determinants are secreted into plant cells.

The Characteristics of Nitrogen Supply in Yellow Soil Planted with Tobacco

LIU Qing-li,REN Tian-zhi,LI Zhi-hong,SHI Jun-xiong,ZHANG Heng,ZHANG Yun-gui

Scientia Agricultura Sinica. 2010, 43(1): 87-95 . doi:10.3864/j.issn.0578-1752.2010.01.011

Abstract ( 987 )

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【Objective】 In this paper, the characteristics of nitrogen mineralization and nitrogen supply of yellow soil planted with tobacco during the growing period of flue-cured tobacco were studied, thus providing a theoretical basis for nitrogen nutrition control of flue-cured tobacco. 【Method】 The field plot experiment adopted the in situ incubation and two experimental fields of yellow soil with different organic matter content were selected. Each experimental plot contained four treatments, i.e. fallow, planting tobacco without N, planting tobacco and supplied 90 kg N?hm2, planting tobacco, and supplied 120 kg N?hm-2. 【Result】 The results showed as follows. In flue-cured tobacco growth period, two peaks of soil nitrogen mineralization were observed, one was at the flue-cured tobacco rapid growing stage (42 days after transplanting), and the second was at the stage of top pruning (77 days after transplanting). During the growth period of the flue-cured tobacco, with the advance of time, the cumulation of the mineralizable N increased, until about 91 days after transplanting of the flue-cured tobacco, the cumulative amount of mineralizable N tended to slow. The effect of soil organic matter content on the dynamics of soil nitrogen mineralization rate was not significant, the use of nitrogen fertilizers resulted in the large fluctuations of the dynamics of soil nitrogen mineralization fluctuations. The soil mineral nitrogen was net fixed at early growth stage of flue-cured tobacco and net mineralization had a significant effect at the late growth stage. 【Conclusion】 Therefore, controlling soil nitrogen mineralization during the late growth period of tobacco can reduce soil nitrogen supply at late growth period of tobacco, at the same time, reduce the inorganic soil nitrogen fixation at the early growth stage of tobacco and promote the pre-release of inorganic nitrogen at the early growth stage of tobacco, and to a certain extent, can also alleviate the problems of excessive soil nitrogen supply at the late growth stage of tobacco.

Effects of Land Use on Heavy Metal Accumulation in Soils and Source Analysis

BAI Ling-yu,ZENG Xi-bai,LI Lian-fang,PEN Chang,LI Shu-hui

Scientia Agricultura Sinica. 2010, 43(1): 96-104 . doi:10.3864/j.issn.0578-1752.2010.01.012

Abstract ( 1163 )

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【Objective】 Heavy metals accumulation and their influential factors were studied in different land use soils, which will provide a theoretical basis for controlling the content of heavy metals in soils. 【Method】 To identify the effects of land use on the accumulation of heavy metals in soils, 148 samples were collected from four land use types including greenhouse, open vegetable field, maize field and forest land in Siping area, Jilin Province, and Cr, Ni, Cu, As, Cd, Pb, and Zn contents of those samples were determined with ICP-Mass. 【Result】 The result showed that there was a rather large difference in effects of the accumulation of Cr, Ni, Cu, As, Cd, Zn in soils under different land use types, except Pb. Based on the assessment compared with background concentrations in soil, the higher accumulation of heavy metals was found in greenhouse and open vegetable field, lesser in maize field and forest field. The order of mean contents of heavy metals in soils was greenhouse＞open vegetable field＞maize field＞forest field. Cd , Cu had relatively serious accumulation in soils compared with Cr, Ni As and Zn. The mean content of Cd in greenhouse was 0.467 mg?kg-1, which exceeded the Grade II of Chinese Soil Quality Criterion GB15618-1995 (6.5＜pH＜7.5) for Cd standard of 0.3 mg?kg-1, and was 5.2 times of Cd in forest field. The mean contents of Cr, Ni, Cu, As, Pb, Zn in soils under four land use types were lower than gradeⅡ of Chinese Soil Quality Criterion. Compared with the age of land use, agricultural chemical compounds application (especial the quality and quantity of applied fertilizer) was the important reason for leading to difference in accumulation of heavy metals in soils under the studied land use types. 【Conclusion】 Heavy metals (Cr, Ni, Cu, As, Cd, Zn) accumulation in soils was significantly affected by land use types, soil in greenhouse accumulates more heavy metals than others. It is suggested that the application of high contents of heavy metals in chemical fertilizer, organic fertilizer and pesticides should be avoided to prevent the accumulation of heavy metal and keep high quality soils for sustainable use.

Construction of A Normal Temperature Straw-rotting Microbial Community and Its Character in Degradation of Rice Straw

LIU Chang-li,WANG Xiao-fen,GUO Peng,LI Pei-pei,SHEN Hai-long,CUI Zong-jun

Scientia Agricultura Sinica. 2010, 43(1): 105-111 . doi:10.3864/j.issn.0578-1752.2010.01.013

Abstract ( 921 )

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【Objective】 In order to investigate the degradation activity, optimal condition of secreted cellulase activity and compose, the microbial community with efficient cellulose degrading ability in 28℃ was studied. 【Method】 Microbial community came from rotted rice straw was enriched and directional domesticated by improved Mandels medium. The standard cellulase activity assays were used to determine cellulase activity, the fermented juice was analyzed by gas chromatography mass spectrometry (GC/MS) method, and denaturing gradient gel electrophoresis (DGGE) was used to identify the composition dynamic of the community. 【Result】 The results showed that the microbial community could degrade 39.6% of rice straw gross weight within five days. When the culture medium volume was 1/5 of the whole and pH at 6 on the 5 days culture, the CMC reached the highest of 14 IU?ml-1. During the rice straw degradation, more than ten kinds of products were detected using GCMS. DGGE detected the dynamic change of microbial community composition, and the microbial composition changed greatly in different periods. The phylogenetic tree derived from 16s rDNA sequence was found that the closest relatives belong to Clostridium sp., Brevibacillus sp., Bartonella sp.、Bacteroidetes sp. 【Conclusion】 This microbial community could accelerate rice straw rotting.

Mapping and Analyzing QTLs of Yield-Associated Agronomic Traits of Greenhouse Cucumbers

CHEN Qing-jun,ZHANG Hai-ying,WANG Yong-jian,LI Wan-yu,ZHANG Feng,Mao Ai-jun,CHENG Ji-hong,CHEN Ming-yuan

Scientia Agricultura Sinica. 2010, 43(1): 112-122 . doi:10.3864/j.issn.0578-1752.2010.01.014

Abstract ( 1126 )

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【Objective】 Autumn-winter and winter-spring are two important cropping seasons for cultivating cucumber (Cucumis sativus L.) in solar greenhouses in China. This study was made to map and analyze some candidate QTLs of yield-associated agronomic traits in cucumber, so as to provide information for molecular marker assisted selection of yield of greenhouse cucumber. 【Method】 A population of 113 cucumber recombinant inbred lines (RILs) derived from a cross between Europe 8 and Qiupeng was used as materials in this study. Based on the RIL molecular linkage map consisting of 182 markers constructed previously,candidate QTLs conferring 9 yield-associated traits were analyzed. 【Result】 A total of 58 QTLs were detected in this study. Among them, 1 QTL conferring single plant yield（SPY）was located on the linkage group LG4; 6 QTLs conferring daily gain weight (DAW) were located on LG2,LG3 and LG6; 5 QTLs conferring single fruit weight (SFW) were located on LG1 and LG5; 2 QTLs conferring the number of fruit setting（NFS）were located on LG2 and LG4; 1 QTL conferring fruit abortion rate (FAR) was located on LG7; 28 QTLs conferring first female flower node (FFN) were located on LG1 through to LG8; 8 QTLs conferring total number of leaves (TNL) were located on LG2, LG7, and LG4; 2 QTLs conferring leaf areas (LA) were located on LG1 and LG3. The above-mentioned QTLs were detected in only one cropping season. There were 5 QTLs conferring female flower number (FFN) on LG2, of which, ffn2a and ffn2b, were detected in both cropping seasons, and with the same positive genetic effects. Some concentrated regions and cluster distribution of QTLs were also found in this study. 【Conclusion】A total of 58 candidate QTLs in a greenhouse-cultivated cucumber population were detected in the study. The two QTLs, ffn2a and ffn2b, were found to be expressed consistently under both cropping seasons of the greenhouse cultivation environment.

Photosynthetic Characteristics of Pepper Leaves Under Low Temperature, Weak Light and Salt Stress

ZHANG Zhi-gang,SHANG Qing-mao

Scientia Agricultura Sinica. 2010, 43(1): 123-131 . doi:10.3864/j.issn.0578-1752.2010.01.015

Abstract ( 1013 )

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【Objective】 The aim of this study was to investigate the response mechanism of photosynthetic characteristics in pepper (Capsicum annuum L.) leaves subjected to low temperature, weak light and salt stress, and to offer a theoretical basis for the pepper production in protected culture in winter and spring. 【Method】 Chlorophyll content, photosynthetic parameters and chlorophyll fluorescence parameters in pepper leaves were determined. 【Result】 Compared with the control, contents of Chl a, Chl b and Chl(a+b) in leaves treated with weak light, weak light under salt stress increased by 25.91%-35.75%, 31.34%-44.78%, and 27.31%-38.08%, respectively, while other treatments were reverse. Moreover, net photosynthetic rate, stomatal conductance, transpiration rate, the maximal efficiency of PSⅡ photochemistry (Fv/Fm), quantum yield of PSⅡ electron transport (ΦPSⅡ) decreased and antenna heat dissipation (D), excess light energy (E) of PSⅡ increased under single low temperature, weak light, salt stress or combined stress, especially low temperature under salt stress. 【Conclusion】 Although net photosynthetic rate decreased under single or combined stress, the limited factor of different stresses leading to decreased net photosynthetic rate in pepper leaves is different. The effects of low temperature under salt stress on photosynthetic characteristics in pepper leaves are the greatest and the lowest is weak light. With the effect of low temperature, weak light and salt combined stress is the next to low temperature under salt stress.

Judgement Parameters of Canopy Overcrowed for Reformative High Trunk Open Centre Shape of Fuji Apple with Standard Rootstock

SHANG Zhi-hua,WEI Qin-ping,SUN Li-zhu,WANG Xiao-wei,ZHANG Qiang,FU Li-hua

Scientia Agricultura Sinica. 2010, 43(1): 132-139 . doi:10.3864/j.issn.0578-1752.2010.01.016

Abstract ( 925 )

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【Objective】 The aim of the present experiment is to study the judgment parameter of canopy overcrowed for reformative high trunk open centre shape of Fuji apple with standard rootstock. 【Method】 The canopy was divided into 50 cm×50 cm× 50 cm and the relative light intensity, type and number of shoots, yield and quality of fruit were measured in different layers and positions of the canopy with twenty-year-old reformative high trunk open centre shape of Fuji apple tree in different growing seasons. 【Result】 The distribution of relative light intensity in the canopy was gradually descended from top to bottom in different layers, and was gradually increased from inner to outer of the canopy in the same layers. The relative light intensity below 2.5 m position of the canopy were less than 31.48%. In June when the leaf area first formed, the percent proportion of ＜30% relative light intensity in volume of the canopy was 47.62%. The yields mainly located in 1.5-2.0 m height of canopy and were calculated by 6.32×104 kg?hm-2. The branches(shoots) mainly distributed below 2.5 m height of the canopy and was 82.92% of total branches(shoots). The percentage of the branches(shoots) in outer, middle and inner positions of canopy was 53.08%, 36.92%, and 10.00%, respectively. The amount of total branches(shoots) and ratios of long, medium, spur shoots were 1.21×106 bars/hm2, 7.97%, 53.63% and 38.41%, respectively, in overcrowed canopy. 【Conclusion】 There are three judgment parameters of reformative high trunk open centre shape of Fuji apple that the percentage of ＜30% relative light intensity in volume of the canopy were 47.62% , total branches(shoots) were 1.21×106 bars/hm2 and shoots ratio in inner of canopy was ＜10% .

Effects of Intercropping Aromatic Plants on Soil Microbial Quantity and Soil Nutrients in Pear Orchard

Scientia Agricultura Sinica. 2010, 43(1): 140-150 . doi:10.3864/j.issn.0578-1752.2010.01.017

Abstract ( 988 )

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【Objective】 This paper was aimed to discuss the effects of intercropping aromatic plants on the microbial quantity and nutrients in sand soil in pear orchard. 【Method】 Intercropping Centaurea cyanus, Saturela hortensis L, Nepeta cataria, Agerarum houstonianum and Ocimum basilicum L were used as experimental treatments and natural grass was used as control to study the soil microbial quantity , the content of the soil nutrients,as well as the relationship among them during their young fruit period,fruit enlargement period, and fruit maturity period, respectively. 【Result】 Compared with the control, intercropping Centaurea cyanus and Ocimum basilicum L increased the microbial quantity significantly in different developing periods and at each soil layer , as well as regulated the ratio of the soil fungi, bacteria and actinomyees in soil microbial community. Intercropping aromatic plants also showed different effects on microbial quantity and ratio. The soil microbial quantity under different treatments was more than that in young fruit period, and the ratio among the fungi, bacteria and actinomyees in fruit enlargement and maturity periods was more favorable than the latter in different layers. The microbial quantity was much higher at layer of 0-20 cm than that of 20-40 cm under different treatments. Compared with the control, all treatments showed different effects on soil nutrient. The content of the soil nutrients under different treatments showed various trends with the change of developing period. The correlation analysis showed that there were multiple significant positive correlations between microbial quantity and the content of soil nutrients with aromatic plants intercropping in sand soil in pear orchard. 【Conclusion】 From analysis it could be seen that intercropping aromatic plants in sand soil in pear orchard had good regulatory effects on the soil microbial quantity , the ratio of the soil fungi, bacteria and actinomyees in soil microbial community and the soil available nutrients, among which, intercropping Ocimum basilicum L on the soil microbial quantity and content of the soil nutrients was better than other treatments.

Degradation of Chlorpyrifos and Fipronil in Rice from Farm to Dining Table and Risk Assessment

ZHANG Cun-zheng,ZHANG Xin-ming,TIAN Zi-hua,HE Dan-jun,LIU Xian-jin

Scientia Agricultura Sinica. 2010, 43(1): 151-163 . doi:10.3864/j.issn.0578-1752.2010.01.018

Abstract ( 1118 )

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【Objective】 Degradation of pesticide residue (chlorpyrifos and fipronil) in rice from farm to dining table and risk assessment for human health were studied to reveal the magnitude of the risks faced by different populations having interest in rice, and to provide a guiding line for application of pesticide in field, and to refine and update the human health risk assessment data while helping to determine a MRL value and harvest interval. 【Method】 A field trial was designed following the protocol of NY/T788-2004, and residues in harvested rice were monitored by GC. Residue levels of post-harvest-applied chlorpyrifos and fipronil during storage, exposure to sunlight, washing and cooking processes (boiled rice) of brown rice were investigated; Several perimeters of evaluation model of dietary exposure evaluation model (DEEM) was employed to estimate the acute and chronic risk faced by different populations of interest, POR (percent of RfD ) and MOE (Margin of the exposure) were included. 【Result】 A positive correlation was found in pesticide residue betweon harvested rice and treatment of pesticide in field, residue presented in harvested rice raised with the raising application rate and treatment times. The concentration of pesticide residue decreased as a function of time during rice storage and preparing and cooking processes. The pesticide residues decreased by 91.16% and 96.16% at the end of the experimental period. The boiling process had a significant effect on the major degradation of pesticides. Risk assessment to human health showed that chlorpyrifos residue in rice should be concerned, and dietary exposures from eating food crops treated with chlorpyrifos exceeded the level of concern for the entire population. Chronic dietary risk from food alone showed that dietary exposures from eating food rice treated with fipronil was below the level of concern for the entire population, including children. Acute dietary (food only) risk estimate for chlorpyrifos as percent of aPAD (acute population adjusted dose) over 100% frequently when the treatment rate and times raised, the result showed that risk faced by boys under the age of 14 was higher than the girls at the same age, and the risk level reversed for the subpopulation at the age above 14. The risk faced by population of rural residential region was more serious than the risk faced by population of urbanite with rice as thceir food, the most sensitive population subgroup, children and male in population of countryside faced higher acute dietary risk than the other subpopulation groups. Thus acute dietary risks from food (rice) alone are of concern. Re-identification of harvest interval found to be the critical point and most important measure to mitigate risk to all population for rice safe eating. The results showed that all risk level of trials for human health decreased to acceptable range when the harvest interval extended to 14 days. 【Conclusion】 Risk quotients indicate that multiple application and double rate of chlorpyrifos increase the risks to entire population and prolong exposures to toxic concentrations. To address these risks, a number of measures including reduced application rates (should not be doubled at single application), increased retreatment intervals (longer than 7 days), extended interval of harvest (at least 14 days) will be needed. Concerning about the toxicity of fipronil to fish, bees and aquatic invertebrate specie, MRL of fipronil in rice is suggested being 0.01 mg?kg-1, which followed the Codex regulation.

Studies on the Secondary Structure and Heat-Induced Gelation of Pork Myofibrillar Proteins as Affected by pH

FEI Ying,HAN Min-yi,YANG Ling-han,ZHOU Guang-hong,XU Xing-lian,PENG Zeng-qi

Scientia Agricultura Sinica. 2010, 43(1): 164-170 . doi:10.3864/j.issn.0578-1752.2010.01.019

Abstract ( 1004 )

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【Objective】 The objective is to study the effect of pH on the secondary structure, α-helix of pork myofibrillar protein, and the hardness, water holding capacity (WHC) and microstructure of its heat-induced gel. 【Method】 The α-helix of pork myofibrillar protein under different pH were measured by circular dichroism (CD), and the hardness, WHC and microstructure of the heat-induced gel were determined by texture analyzer, centrifugation method and scanning electron microscope (SEM), respectively. 【Result】 The α-helix of pork myofibrillar protein and WHC of its heat-induced gel increased with pH away from pI, the hardness reached its maximum when pH was 6.0, the gel had a uniform and orderly microstructure in neutral, while it had disorderly and uneven microstructure with polymer in the acidic. 【Conclusion】 The α-helix of myofibrillar protein is positive correlated with the WHC of its heat-induced gelation, and the gel has an orderly microstructure with more α-helix, while rough microstructure with less α-helix.

Expression of Nitric Oxide Synthases in Different Coat Color of Alpaca Skin Hair Follicle

CAO Jing,DONG Chang-sheng,HE Xiao-yan,DONG Yan-jun,FAN Rui-wen,HE Jun-ping,WANG Hai-dong,YU Xiu-ju

Scientia Agricultura Sinica. 2010, 43(1): 171-175 . doi:10.3864/j.issn.0578-1752.2010.01.020

Abstract ( 1095 )

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【Objective】 The experiment was made to study the relation of alpaca coat color and the expression of nitric oxide synthases (NOS) in alpaca skin. 【Method】 The relationship between the alpacas coat color and the expression of NOS was studied by determining the expression and localization of NOS in alpaca skin with different hair colors. The protein expression of NOS was determined by western blotting. The tissue localization of NOS was examined by immunohistochemistry. 【Result】 There was a positive expression of three NOS in skin of white alpaca group and brown alpaca group, the expression in hair follicle tissue significantly higher than peripheral connective tissue (P＞0.05). In hair follicle tissue of white alpaca group and brown alpaca group, the expression of NOS1 and NOS3 showed a weak positive, NOS2 showed a strong positive. The expression of NOS2 in hair follicle tissue of white alpaca group was significantly lower than those in brown alpaca group (P＞0.05). The expression of NOS1 and NOS3 showed no significant difference in hair follicle tissue of white alpaca group and brown alpaca group (P＜0.01). The expression of three NOS showed no significant difference in connective tissue (P＞0.05). Western blotting showed that the expression of NOS2 in white alpaca group was significantly lower than those in brown alpaca group (P＜0.05).【Conclusion】This is the first study on the expression of NOS in alpaca skin with different coat colors, three NOS main expression in hair follicle site. The expression of NOS1 and NOS3 is not related with coat color; the expression of NOS2 in hair follicle is related with coat color. The experiment has laid a foundation for further study on the effect of nitric oxide signal on melanin production.

Proteomic Analysis of the Honeybee (Apis mellifera L.) Caste Differentiation Between Worker and Queens Bees Larvae

WU Jing,LI Jian-ke

Scientia Agricultura Sinica. 2010, 43(1): 176-184 . doi:10.3864/j.issn.0578-1752.2010.01.021

Abstract ( 1141 )

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【Objective】 To investige into the differentially expressed proteins between the honeybee (Apis mellifera L.) worker and queens larvae , the proteome profiling of the workers and the queens were compared on day 3, day 5 and day 11 during the developmental process of the honeybee caste differentiation. 【Method】 Two-dimensional gel electrophoresis (2-DE) was employed to set up protein profiling of the queens and the workers during their eleven days development. Then the protein abundance, molecular weight and pI were analyzed. 【Result】 Totally, 288 and 274 proteins were detected in the proteome profile of queens on day 3 and day 5, respectively, which is significantly higher than those detected in workers ( 259 and 236, accordingly). And in the head of 11-day pupae, also evidently higher number of total proteins (311) was detected in queens than those of workers (278). On the third day, 156 proteins were presented in both queens and workers larvae, while 132 proteins were specific to queens larvae, and 103 proteins were exclusive to workers larvae. While on the fifths day, larvae of queens and workers had 95 shared proteins, and 179 proteins were specific to queen larvae and 141 proteins to workers larvae. Until the eleventh day, 194 proteins were resolved in the head of queen and workers, while 117 and 84 proteins were unique to the heads of queen and workers, respectively.【Conclusion】 Significant differences were found in the proteome between queens and workers during the process of caste differentiation. While the increased number of total and specific proteins in queens showed that the metabolic rate and gene expression of queens is higher than those of workers. The proteins both in queens and workers suggest that they should be indispensable for caste development, but their expression patterns are different between two bee castes. The protein specifically expressed at each time-point indicate that different caste developmental stages need specific proteins to be involved in.

Development of a Novel RT-LAMP Assay for Porcine Reproductive and Respiratory Syndrome Virus

XIN Ting,HOU Shao-hua,JIA Hong,GUO Xiao-yu,DING Jia-bo,LI Yan-peng,DING Min,ZHU Hong-fei

Scientia Agricultura Sinica. 2010, 43(1): 185-191 . doi:10.3864/j.issn.0578-1752.2010.01.022

Abstract ( 983 )

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【Objective】 The objective of the experiment is to develop a one-step reverse transcription loop-mediated isothermal amplification (RT-LAMP) assay for rapid and sensitive detection of porcine reproductive and respiratory syndrome virus (PRRSV). 【Method】 Three pairs of primers were designed to identify 8 positions in N gene, the sensitivity and specificity of RT-LAMP were determined and clinical test was performed under optimized amplification condition. 【Result】 A RT-LAMP assay was developed for the rapid detection of PRRSV. The process of assay was conducted with one step amplification within 70 minutes and amplification results were visualized. The sensitivity of RT-LAMP assay was 10 000 times higher than RT-PCR. In the detection of 50 porcine serum samples, this assay showed excellent agreement with the standard RT-PCR assay (κ＝0.83). 【Conclusion】 Therefore, the rapid and simple assay is a potential useful technique for PRRSV detection in field condition.

Cloning, Expression of Cysticercus Cellulosae dUTPase and the Determination of Its Enzymatic Activity

LIU Zhen-yong,Lü Zhi-hui,ZHENG Ya-dong,DOU Yong-xi,QIAO Jun,LUO Xue-nong,ZHANG Yan,JING Zhi-zhong,CAI Xue-peng

Scientia Agricultura Sinica. 2010, 43(1): 192-199 . doi:10.3864/j.issn.0578-1752.2010.01.023

Abstract ( 887 )

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【Objective】 In order to develop new drugs against cysticercosis, the dUTPase gene from Cysticercus cellulosae was cloned and expressed. Functionality of the purified recombinant dUTPase was proven by demonstrating catalytic activity towards different substrates. 【Method】 The gene of the dUTPase was obtained by reverse transcription-polymerase chain reaction (RT-PCR) from Cysticercus cellulosae, then cloned into fusion expression vector of pET, and highly expressed in E. coli BL21. The His6-tagged recombinant dUTPase was purified on Ni2+–IDA–Sepharose, and the enzymatic activity was investigated. 【Result】 Analysis showed that the open reading frames (ORFs) of dUTPase genes from both cysticerci and oncospheres shared 100% identity, and five conserved motifs were also found in amino acid sequence. The molecular mass of recombinant dUTPase was 21 kD as judged by SDS-PAGE. After purified, the protein concentration of dUTPase was 2.863mg?mL-1. The analysis of the enzymatic activity indicated that the recombinant dUTPase could catalyse the hydrolysis of dUTP and the activity of enzyme was enhanced by Mg2+ and inhibited by EDTA. 【Conclusion】 The dUTPase from Cysticercus cellulosae has been cloned, expressed and identified, which provided a material foundation for a novel drug design against cysticercosis.

The Ultrastructure of Taenia solium Oncosphere Membrane and Egg

SUN Xiao-lin,CAI Xue-peng,MI Xiao-yun,CHEN Huai-tao

Scientia Agricultura Sinica. 2010, 43(1): 200-205 . doi:10.3864/j.issn.0578-1752.2010.01.024

Abstract ( 1282 )

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【Objective】 The ultrastructure of oncosphere membrane and egg of Taenia solium were observed in a study. 【Method】 The decoction areca and pumpkin seeds were taken by the patients (limosis) to get T. solium mature eggs. After being treated with sodium hypochlorite to break the eggshells, the oncospheres were gathered with isoosmotic percoll solution and were treated with the artificial intestinal juice to activate oncospheres. Then the ultrastructure of egg and oncosphere membrane were observed by scanning electron microscope (SEM) and transmission electron microscope (TEM). 【Result】 The egg was in oval shape with the size of 50-58 μm and consist of embryophore, inner cytoplasmic layer, oncosphere membrane and oncosphere. The structure of developmental T. solium oncosphere membrane was composed of residual layer of outer envelope, outer cytoplasmic layer (including the outer irregular compact layer and the inner homogeneous lucent layer), the embryophore (including blastoderm layer and inner cytoplasmic layer) and oncosphere membrane from outside to inside, respectively. However,there was only one layer in the mature oncosphere membrane with a size of 49-51 nm. 【Conclusion】 The ultrastructure of the oncosphere membrane and egg of T. solium are similar to that of Hymenolepis diminuta and T. saginata except that the envelope is not present around the glycogen granule in underlayer of oncosphere membrane of T. solium.

Coupling Effect of Water and Nitrogen on Population Physiological Indices Under Alternative Furrow Irrigation

LI Pei-ling,ZHANG Fu-cang,JIA Yun-gang

Scientia Agricultura Sinica. 2010, 43(1): 206-214 . doi:10.3864/j.issn.0578-1752.2010.01.025

Abstract ( 900 )

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【Objective】 The objective of this paper is to study the coupling effect of water and nitrogen on population physiological indices, cotton biomass and cotton yield under alternative furrow irrigation (AFI). 【Method】 The alternate furrow irrigation methods was used, the second common rotary combination design water and nitrogen application rate, and the cotton population physiological indices changes in different water and nitrogen combinations were determined in test area of field. 【Result】 The result showed that a single-factor effect of water or nitrogen on cotton physiological indices, cotton yield, cotton biomass was observed. Leaf area duration (LAD), crop growth rate (CGR), net assimilation rate (NAR), leaf area index (LAI), yield, biomass of the irrigation and nitrogen single-factor effect indicates that when nitrogen rate was at 56.2-122.8 kg?hm-2, population physiological indices, yield, biomass and nitrogen application rate were significantly and positively correlated. Nitrogen application rate at 122.8-134.2 kg?hm-2, LAD and CGR were significantly and positively correlated, and NAR, LAI, biomass, yield, and had no significant changes. When irrigation norm was at 37.52-192 mm, population physiological indices, cotton yield, cotton biomass and irrigation had a significant positive correlation. When irrigation norm was at 192-218.48 mm, LAD, CGR, LAI and irrigation were significantly correlated, NAR, and biomass, cotton yield did not change significantly. Cotton population physiological indices, cotton biomass, cotton yield of water and nitrogen coupling effect show that when irrigation norm was at 37.52-192 mm, LAD, CGR, NAR, and LAI increased significantly with the increase of irrigation norm, but the increasing trend with the increase in nitrogen application rate decreased. Irrigation in the 192-218.48 mm, under different nitrogen application rate, population physiological indices, biomass, yield no significant change. When the nitrogen rate was at 56.2-95.2 kg?hm-2, LAD, CGR, NAR, and LAI increased significantly with the increase of nitrogen application rate, but the increasing trend with the increase of nitrogen application rate tended to decrease. When nitrogen application rate was 95.2-134.2 kg?hm-2, the population physiological indices, cotton biomass, cotton yield had no significant changes. 【Conclusion】 The coupling effect of water and nitrogen on cotton physiological indicators, biomass and yield under alternative furrow irrigation, can be utilized which is to coordinate cotton growth and development, thus improving cotton yield and water and nitrogen use efficiency.

Analysis of Genetic Relatedness of Genetic Resources of Tagetes as Revealed by ISSR

ZENG Li,ZHAO Liang-jun,SUN Jia,ZHAO Zi-gang,YANG-fan

Scientia Agricultura Sinica. 2010, 43(1): 215-222 . doi:10.3864/j.issn.0578-1752.2010.01.026

Abstract ( 955 )

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【Objective】 ISSR markers were used to study the genetic relationship and investigate the genetic diversity and genetic structure for providing a theoretical basis and technical support for appropriate conservation and application of existing genetic resources of Tagetes. 【Method】 Genetic relationship of 29 accessions of Tagetes erecta and 2 accessions of T. patula were analyzed using ISSR markers. 【Result】 The genomic DNA of 31 accessions of genus Tagetes was amplified with 11 primers, the bands produced by each primer ranged from 5 to 11, and 6.8 bands were obtained on average, the polymorphic loci ranged from 40%-100%. The cluster analysis of ISSR data indicated that the resources classified by ISSR was basically in accordance with that by traditional method, the accessions were generally classified into 2 groups of different species, and cultivars of same lines were clustered into a subgroup. 【Conclusion】 The genetic relationship and genetic diversity of genus Tagetes could be assessed efficiently by ISSR markers.

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