Scientia Agricultura Sinica

Comparison among Gene Supervised Clustering Methods for DNA Microarray Expression Data

Scientia Agricultura Sinica. 2007, 40(10): 2119-2127 .

Abstract ( 1242 )

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Several typical supervised clustering methods, Gaussian mixture model-based supervised clustering (GMM), K-Nearest-Neighbor (KNN), binary support vector machines (SVMs) and multicategory support vector machines (MC-SVMs), were employed to classify the computer simulation data, yeast cell cycle microarray data and 60 human cancer cell lines (NCI-60) microarray data. False positive, false negative, true positive, true negative and clustering accuracy were compared among these methods. The results are as follows. (1) For classify thousands of gene expression data, the performances of two GMM methods have the maximal clustering accuracy and the least overall FP+FN error numbers based on the assumption that the whole set of microarray data is a finite mixture of multivariate Gaussian distributions. Furthermore, when the number of training sample is very small, the clustering accuracy of GMMⅡ method have superiority over GMMⅠ method. (2) In the general, the superior classification performance of the MC-SVMs are more robust and more practical, which are less sensitive to the curse of dimensionality and not only next to GMM method in clustering accuracy to thousands of gene expression data, but also more robustness to a small number of high-dimensional gene expression samples than other techniques. (3)Among MC-SVMs, in case of large sample sizes, OVO and DAGSVM perform better; In case of moderate sample sizes, five MC-SVMs methods perform very similar; Otherwise, OVR, WW and CS yield the better results when sample sizes are small. (4) A suggestion for the supervised clustering microarray data is that one should consider the data feature and experiment when choose an appropriate method. Two kinds of these methods should be trial calculation to obtain better clustering result.

Stable expression of QTL for milling quality of rice (oryza stativa L.) using CSSL population

Scientia Agricultura Sinica. 2007, 40(10): 2128-2135 .

Abstract ( 1190 )

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A chromosome segment substitution line (CSSL) population, derived from the cross of Asominori and IR24 with Asominori as the recurrent parent, was planted and phenotyped for brown rice rate (BR), milled rice rate (MR) and head rice rate (HR) in four sites. QTL for milling quality were characterized and expression stability of these QTL was further investigated by using target CSSLs harboring certain QTL. The phenotypic values of these three traits all showed continuous distributions and transgressive segregations in the CSSL population across the four environments.. Meanwhile, a total of 30 QTL were identified for the three quality traits, among which four QTL, qMR-6、qMR-8、qHR-3 and qHR-6, were all consistently detected in the four different environments. Moreover, phenotypic values were different significantly (P<0.05) between Asominori and the target CSSLs harboring any of the four QTL alleles, indicating expression stability of the four QTL was relatively higher. In addition, QTL qMR-8、qBR-8b and qHR-8 were co-located in the G1149-R727 interval on chromosome 8 carrying a QTL cluster. Interestingly, many major QTL affecting rice eating, cooking, appearance and nutrient quality was also mapped into the QTL cluster in our previous studies. These results thus obtained provide us a good chance to elucidate the complex mechanism that formation of many rice quality traits was simultaneously controlled by a gene or a gene cluster on chromosome 8.

Chromosomal Localization of Genes Associated with

Scientia Agricultura Sinica. 2007, 40(10): 2136-2144 .

Abstract ( 1051 )

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[Objective]Chromosome substitution lines between Chinese spring and Synthetic 6x under different water treatments were used to locate the gene controlling spikelet progress and drought tolerance.[Method]A series of spikelet differentiation progress were observed.[Result] The result indicated that spikelet progress of various substitution lines and that of parents, including spike and flower anlage periods, were almost alike. Under normal irrigation,2B substitution line is the latest in the progress of spikelet differentiation, 7D substitution line is the second, while Chinese spring and other substitution lines are the rapidest. The spikelet differentiation progress of 2D substitution line is much slower than Chinese spring and other substitution lines under drought condition.[Conclusin] It is concluded that the genes inhibiting spikelet differentiation may be located on 2B and 7D chromosome, and that the genes of drought tolerance may be located on 2D chromosome. The results can provide theoretic basis for the genetic and breeding research concerned with drought resistance and water saving.

Studies on selecting indices and evaluation methods for screening waterlogging tolerance

Scientia Agricultura Sinica. 2007, 40(10): 2145-2152 .

Abstract ( 1219 )

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: 【Objective】Waterlogging stress is one of the major problems in barley production especially in wetlands. Development of tolerant cultivars is the most cost efficient and environmentally safe way to overcome this problem. The objective of this study was to investigate suitable selection criteria for waterlogging tolerance in barley. 【Method】 165 DH lines derived from a barley cross（Yerong × Franklin) were analysed using principal component analysis, dynamic clustering and subordinate function based on the specimen correlation matrix of 12 traits, including plant height(PH), spike length (SL), the top 1st internode length(TFIL), spike per plant (SPP), grains per spike(GPS), grain weight per plant(GWPP), grains per plant(GPP), straw weight per plant(SWPP), weight of 1000 grains(WTG), number of green leaf (NGL) and Chlorophyll 1-2(Chl 1-2). 【Results】Three principal components and their functional equations, two for spike-grain factor and one for NGL factor, were used to identify lines for waterlogging tolerance, which led to the classification of 165 lines into 3 different levels of tolerance. 30, 93 and 42 lines were classified as waterlogging tolerant, medium tolerant and sensitive, respectively. 【Conclusion】The new selecting indices and comprehensive evaluation method provides a useful tool for further study on QTLs controlling waterlogging tolerance and selecting for waterlogging-tolerance in a breeding program.

Assessment of Genetic Diversity among Tobacco Germplasms by ISSR Markers

Scientia Agricultura Sinica. 2007, 40(10): 2153-2161 .

Abstract ( 1471 )

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【OBJECTIVE】Assessing the genetic diversity among tobacco accessions and analyzing the genetic relationships between various types of tobacco provide the essential information for the mining and utilization of tobacco germplasms;【METHOD】One hundred and nineteen tobacco accessions including various types of tobacco were used for ISSR analysis. The genetic similarity coefficients were estimated and cluster analysis was carried out using UPGMA method;【RESULTS】Six hundred and seventy-two bands, all of which were polymorphic, were amplified using 21 ISSR primers. Out of them, one hundred and sixteen bands were specific for N. tabacum. Pairwise genetic similarity coefficients among accessions in N. tabacum ranged from 0.779 to 0.945, with an average of 0.868. Among the flue-cured tobacco varieties, genetic similarity coefficients ranged from 0.812 to 0.933, with an average of 0.876. The tobacco accessions could be clustered into the different subgroups or classes according to the types of tobacco. The flue-cured tobacco accessions introduced from abroad were obviously not separated from the domestic accessions including landraces and varieties developed in China. There are low genetic similarity coefficients between the accessions in N. tabacum and other tobacco species. The species N. tabacum and its progenitor N. sylvestris were clustered into the same class, and N. longiflora and N. plumbaginifolia, both of which belong to the group Alatae of the subgenera Petunioides, were clustered into the same class. The results obtained in the ISSR dendrogram agreed with the presumed genetic differentiation;【CONCLUSION】There was low genetic diversity among the flue-cured tobacco germplasms in China. The germplasms clustered into the different subgroups or classes should be utilized in the selection of the parents. In order to widen the genetic base of the flue-cured tobacco varieties, the genetic potentials in the wild tobacco should be explored.

The regulatory mechanism of different nitrogen form on photosynthetic efficiency of rice plants under water stress

Scientia Agricultura Sinica. 2007, 40(10): 2162-2168 .

Abstract ( 1349 )

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【Objective】 In order to study the characteristics of photosynthetic physiology of rice plants under different nitrogen form nutrition and water stress. 【Method】A hydroponic experiment with simulated water stress by PEG(6000) was conducted, the photosynthetic characters and Rubisco content and activity were studied by analyzing carboxylation efficiency (CE), total nitrogen content, soluble protein content and Rubisco content in newly expanded leaf of rice plants. 【Result】 The results showed that, under ammonium nutrition, there was no significant different CE of rice plant between water stress and non-water stress, while Rubisco activity was reduced 13.3% by water stress. However, under nitrate nutrition, both of CE and Rubisco content reduced under water stress, and Rubisco activity reduced 23.0%. Thus, the inhibitory effect on Rubisco activity (CE/Rubisco) of rice plants supplied with ammonium was lower than that of supplied with nitrate. Furthermore, water stress also increased total nitrogen content in newly expanded leaf of rice plants supplied with ammonium, while no effect under nitrate supply. Water stress increased soluble protein content in newly expanded leaf supplied with three nitrogen forms, although no significant effect on the ratio of Rubisco content to soluble protein under ammonium supply, decreased this ratio under nitrate supply. 【Conclusion】Inhibitory effect of water stress on photosynthetic efficiency of rice plants supplied with ammonium was lower than that of supplied with nitrate.

Development of 2-DE protocol for wheat leaf proteomics

Scientia Agricultura Sinica. 2007, 40(10): 2169-2176 .

Abstract ( 1433 )

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【Objective】To develope an efficient and higher resolution 2-DE procedure for future wheat leaf proteome research.【Method】Sample preparation is carried out according to two protocols modified from former researchers,following 2-DE and MS analysis. 【Result】These two protocols presented here are efficient protein extraction methods,and both are compatible with MS analysis.【Conclusion】The results indicate that both protein extraction protocols utilized are useful in wheat leaf proteome research.

Analysis on Activity of Enzymes Associated with Fiber Development in Different Genotypes Cotton and Its Relationship with Fiber Strength

Scientia Agricultura Sinica. 2007, 40(10): 2177-2184 .

Abstract ( 1164 )

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Abstract: 【Objective】To study the dynamics of enzyme activities, which play important roles during cotton fiber development, and its relationship with fiber strength, and provide studying basis for the ways of physiological adjusting to improve fiber strength, 【Method】field experiments were carried out in Nanjing, Jiangsu Province (the Yangtze River region in China) in 2004 and 2005 by choosing three genotype cottons with significant differences in fiber strength as materials. The study focused on the time course of sucrose synthetase, β-1,3-glucanase, peroxidase and IAA oxidase activities during the secondary wall thickening in cotton fiber. The boll samples were collected from the first and second fruiting positions of 6 to 8 sympodial branchs. 【Result】The results showed that β-1,3-glucanase activity continuously decreased during the secondary wall thickening in cotton fiber. The activity curves of other enzymes (sucrose synthetase, peroxidase and IAA oxidase) were all unimodal and they might have a certain synergistic effect during fiber thickening development. However, there were obviously differences among three genotypes in the levels of enzyme activities and occurrence time of the peak. The cultivar (Kemian 1), which produced high strength fiber, had higher peak value of the related enzyme (sucrose synthetase, β-1,3-glucanase, peroxidase and IAA oxidase) activities, and slower descend trend subsequently. These resulted in a longer and more tempered cellulose accumulation, and in the wider range and longer period of fiber strength increase. While the cultivars (Dexiamian 1 and Sumian 15), which produced lower strength fibers, the items were just the contrary. And the cultivar (NuCOTN 33B), which produced medium strength fiber, the items were intervenient.【Conclusion】There are significant differences in the related enzymes activities among different genotypes. And that may be one of important factors which lead to the great difference in the characteristics of cellulose accumulation and the fiber strength formation in different genotype cottons.

Accumulation of Anthocyanin in Developing Root and its Relationships with Major Economic Traits in Purple-Fleshed Sweetpotato ( Ipomoea Batatas (L.) Lam )

Scientia Agricultura Sinica. 2007, 40(10): 2185-2192 .

Abstract ( 1379 )

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Abstract: 【Objective】Accumulation law of anthocyanin in developing root and the relationship between the law and major economic traits in purple-fleshed sweetpotato ( Ipomoea Batatas (L.) Lam ) were studied in present paper. 【Method】The dynamics of 20 kinds of economic traits in 13 purple-fleshed sweetpotato varieties at 20d, 40d, 60d, 80d, 100d, 120d and140d after their transplanting were investigated, these traits included anthocyanin content in root, length of the longest stem, number of base branches, root number, dry matter contents in stem, foliage and root, fresh and dry yields of root, fresh and dry yields of stem, fresh and dry yields of foliage, fresh and dry yields of vine, fresh and dry yields of whole plant, and rations of photosynthate to root, to stem and to foliage. The correlationships of anthocyanin content with other 19 economic traits in varieties and whole developmental stages were analyzed. So as the correlationships of daily growth rate of anthocyanin content with other daily growth rates of 10 kinds of yields. 【Result】 (1) There were three types of anthocyanin accumulation in these purple-fleshed sweetpotatoes, that is slowly increasing, fluctuating and devious ascending. (2) Anthocyanin content had significant negatively correlationships with the number of base branches, fresh and dry yields of root, ration of photosynthate to root，and had significant positive correlationships with dry matter content of root, length of the longest stem, fresh and dry yields of stem,, dry yield of whole plant and ration of photosynthate to foliage, and had no significant correlationships with other economic traits among purple-fleshed sweetpotato varieties. (3) Purple-fleshed sweetpotatoes belonging to different types of anthocyanin accumulation had different responses to the growth of length of the longest stem, number of base branches, fresh and dry yields of root and photosynthate allotments, and had no responses to the growth of other economic traits during whole developmental stages. (4) Because of the significant negative correlationship between daily growth rates of anthocyanin content and dry matter yield of root, the anthocyanin accumulation competed with dry matter accumulation for photosynthate in root, and the competitive relation was resolved in different way in different purple-fleshed sweet potato varieties.【Conclusion】The three types of anthocyanin accumulation in purple-fleshed sweetpotato can be demonstrated by the their different responses to the growth of economic traits , the competitive relations in accumulations of anthocyanin and dry matter in root, and their different settling ways of these competitive relations. The anthocyanin accumulation characteristics will give scientific proofs for the further research, breeding, cultivation and industrialization of purple-fleshed sweetpotato.

Current Progress in the Research on avrBs3/pthA Family Genes of Xanthomonas oryzae

Scientia Agricultura Sinica. 2007, 40(10): 2193-2199 .

Abstract ( 1372 )

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Xanthomonas oryzae pv. oryzae (Xoo) and Xanthomonas oryzae pv. oryzicola (Xooc) cause bacterial blight (BB) and bacterial leaf streak (BLS) in rice (Oryza sativa)，respectively, which constrain production of this staple crop in China and the world. According to bioinformatics of the genome sequence of Xoo and other recent reports, there are 15-30 avrBs3/PthA (avr/pth) genes in Xoo and Xooc. Recent research results suggest that avr/pth genes are not only involved in virulence when there are no corresponding R gene in rice, but also in suppressing plant innate immunity for hypersensitive response, and in avirulence when there are matching R genes in rice. Despite of avr/pth genes existing Xooc, no R genes were found for BLS in rice, assuming that there are defense inhibitors in Xooc. Members of avr/pth family genes are remarkably similar. However, the main difference is in the number and apparent nature of near-identical 102-base-pair direct repeats in the central portion of the coding sequence. In this review, we postulated that the evolution of the avr/pth genes might occur from the simple to the complex. This may elucidate the pathogenicity differentiation of Xoo and Xooc in rice.

The Homology analysis for sequences of 16SrDNA and tuf gene of Phytoplasma from Jujube witches’broom and Wild jujube witches’ broom

Scientia Agricultura Sinica. 2007, 40(10): 2200-2205 .

Abstract ( 1229 )

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【Objective】Jujube witches’ broom (JWB) is a destructive disease caused by a phytoplasma，that happened in the twenty-five provinces(cities) of china, brought largely economic loss. 【Method】Jujube witches’ broom and wild jujube witches’ broom were investigated in Binxian，Yanliang，Wugong，Yangling，Jiaxian of Shaanxi, Cangzhou of Hebei, Xinxiang of Henan. The 16SrDNA and tuf gene were cloned by polymerase chain reaction from samples of JWB and wild jujube witches’ broom, and sequenced.【Results】The 1239bp and 845bp DNA fragement were amplified in DNA samples not only from infected jujube but also from infected wild jujube. it is found that 16SrDNA is 1239bp, tuf gene is 845bp. The Homology analysis for sequences of 16SrDNA and tuf gene shows that pathogens from Shaanxi, Hebei, Henan are same, and belong to 16SrⅤ-B group. Because there are five differential nucleotides in 16SrDNA between JWB and wild jujube witches’ broom , and homology rate of nucleotide sequences of tuf gene is 91%, we can estimate that wild jujube witches’ broom is infected by different strain of JWB pathogen. 【Conclusion】This article reported sequences of JWB and wild jujube witches’ broom phytoplasmas firstly, and differentiate and classify phytoplasma of JWB and wild jujube witches’ broom. which can provide theory for research of the molecular nosogenesis of JWB phytoplasma and genetic basis.

Avctivity and mode of action of azoxystrobin to four plant pathogens

Scientia Agricultura Sinica. 2007, 40(10): 2206-2213 .

Abstract ( 1520 )

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Azoxystrobin inhibited mycelial growth in Colletotrichum capsici, Botrytis cinerea, Rhizoctonia solani and Magnaporthe grisea respectively; it also inhibited conidia germination, conidia production in Colletotrichum capsici, Botrytis cinerea Magnaporthe grisea and sclerotia forming in Rhizoctonia solani. Moreover, it stayed pigment biosynthesis in Colletotrichum capsici and Magnaporthe grisea somehow. SHAM enhanced inhibition by azoxystrobin, especially in those whose mycelia were not sensitive to azoxstrobin, such as Colletotrichum capsici and Botrytis cinerea with the synergistic ratio more than 200. Oxygen consuming test of the mycelia showed that azoxystrobin inhibited all the four fungi’s respiration in the early stages and with the concentration rising up, the effectiveness became better, it was especially significant in Colletotrichum capsici. However, as time went on, the respiration of the mycelia treated with fungicides recovered and even got stronger than the control after 48h, and salicylhydroxamic acid (SHAM) which acted on the alternative oxidase could not inhibit the oxygen consuming. As a result, it seemed that not the startup of the alternative respiration but the other mechanism caused the ineffectiveness.

Identification and Preliminary Analysis of Genetic Diversity of Cenococcum geophilum Fr.

Scientia Agricultura Sinica. 2007, 40(10): 2214-2220 .

Abstract ( 1111 )

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Cenococcum geophilum Fr. (Cg) is an ectomycorrhizal fungus that has a wide host range and distribution. This mycobiont has been suggested to provide isolate-dependant drought protection to fine roots. It lacks asexual or sexual spores and froms abundant sclerotia that serve as resistant structures under environmental stress. The traditional classification method is very difficult to satisfy to the Cg classification and research of genetic diversity, so little is known about its genetic structure at fine scale. We obtained 27 Cg strains isolated from 6 hosts in China and 5 Cg strains introduced from France as our tested materials, and made identification based on morphological charactertics and PCR method. The genetic diversity of 20 Cg strains was estimated using RAPD and PCR-RFLP of the rDNA internal transcribed spacer (ITS) in this study. PCR-RFLP method used universal primers ITS1/ITS4 to amplified the rDNA ITS of mycelium, then amplified products were digested with HinfⅠand MboⅠ. The digested fragments of PCR products were run on 2% agarose gel, showed that there were obvious differences among strains. RAPD method used selected random primer (5'- CGCACCGCAC-3') to amplified genomic DNA of Cg RAPD produced 19 detectable and reliable DNA bands in the 300bp-2000bp size range. According to the number、position and strength of the DNA bands in agar gel, using PopGen Version 1.31 dendrogram analysis software calculated the genetic distance and genetic similarity among all tested strains. According to the genetic distance to constructed phylogenetic tree of 20 strains, the results indicated that a high degree of genetic diversity was observed among the Cg strains from the same or different hosts.

Purification and Comparison of toxicity Proteins among Extracellular and Intracellular by Xenorhabdus nematophila var. pekingensis

Scientia Agricultura Sinica. 2007, 40(10): 2227-2233 .

Abstract ( 1139 )

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【Objective】Toxicity Proteins among Extracellular and Intracellular produced by the strain CB6 of Xenorhabdus nematophila var. pekingensis were purified and compared in order to obtain the basic data of toxicity proteins for further study.【Method】Toxicity protein fractions from extracellular and intracellular products were purified using the technology of ammonium sulfate precipitation, DEAE-Sepharose FF chromatography, Butyl Sepharose FF chromatography, and Sephacryl S-200 HR chromatography. The protein components were analyzed with the technique of native-polyacrylamide gel electrophoresis (Native-PAGE) and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE). 【Result】Two toxicity protein fractions E1 and E2 respectively from extracellular and intracellular products were purified to electrophoretic homogeneity. The growth inhibitory percentages against Helicoverpa armigera Hübner larvae was 62.63％and 97.90% with the concentrations of 2.58μg/m L E1 and 4.21μg/mL E2 respectively. A series of similar liquid chromatograms of E1 and E2 was gained through same purification procedures and parameters. Moreover, they had a similar migration location whether by native-PAGE or by SDS-PAGE. With SDS-PAGE. E1 and E2 have approximate molecular masses higher up 212kD. They still maintained inhibitory toxicity after heated up to 60℃. Staining experiments showed that the toxicity protein was neither lipoprotein nor glycoprotein.【Conclusion】This demonstrated the possibility that the target protein from extracellular section was the same as that from intracellular one.

Effect of long-term application of K fertilizer on spring maize yield and soil K in northeast China

Scientia Agricultura Sinica. 2007, 40(10): 2234-2240 .

Abstract ( 1288 )

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Using the 13-year fixed site field trials located at selected locations in three provinces of northeast China, Liaoning, Jilin and Heilongjiang, effect of K fertilizer use on maize yield and soil K status in the plough layer was studied. Results showed that effect of K application on maize yield was quite different between the three fixed sites. In the field trial in Heilongjiang, significant maize yield response to K was found starting in the first year; in Jilin, the significant yield response to K was found starting in year 3; while in Heilongjiang, the maize yield response to K was not very stable from 1993 to 2005. The ranges of yield response to K fertilizer application in the field trials in Liaoning, Jilin and Heilongjiang were 9.9%-10.3%, 12.6%-13.6% and 17.5%-21.7%, respectively. In all three sites, K balance in soil-crop system was negative in the treatment with K2O application rate at 112.5kg/hm2, and remarkable K accumulation in soil was found when K2O application rate was 225.0 kg/hm2. Application of K fertilizer significantly increased soil available K content, and the increase in slowly available K in soil was not significant. Considering maize yield and income and soil K status, suitable, quantity of K fertilizer (K2O) was 112.5 kg/hm2 under present experimental conditions.

Effect of different cultivation methods and rain intensity on soil nutrient loss from a purple soil

Scientia Agricultura Sinica. 2007, 40(10): 2241-2249 .

Abstract ( 1278 )

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Non-point pollution to the environment originated from agricultural activities has increasingly drawn concern globally. Though numerous studies have been aimed at this subject, those involving effect of cropping patterns on nutrient loss are rare. The influences of rain intensity and cropping patterns on N and P losses were studied using a completed randomized block experiment adopting artificial rains onto the simulated runoff blocks. Results showed that the amounts of both surface and total runoff were coincided with rain intensity, while the underground runoff was negatively correlated with rain intensity. High rain intensity tended to undermine penetration of rain water into soil and thus reduced soil moisture. At same rain intensity, flat cultivation produced maximal surface runoff and soil erosion. Contour cultivation, however, seemed ideal in controlling soil erosion and surface runoff at small to medium rain intensity, but this effect was weakened at heavy rain intensity. Furthermore, it tended to elevate underground runoff that was responsible for main loss of N. It was observed that P was predominantly lost to sediment and highly correlated with rain intensity. At small to medium rain intensity, N was carried away from the plot by runoff, especially by underground runoff. Thus, it is very crucial to make underground runoff under control in order to control N loss to environment. As for P, any methods to effectively stop soil erosion are the solutions. Under the conditions of this experiment, about 1% of fertilizer N (available N) was leached out of the soil and became the non-point pollutant to environment; while for P, its loss to environment was very minor, only accounting for less than 0.3‰ to 1‰ of fertilizer P applied.

Simulating Temporal-Spatial dynamics of Greenhouse Gas Emits From Rice Paddy Field in Liaodong Costal Region

Jiaguo Qi

Scientia Agricultura Sinica. 2007, 40(10): 2250-2258 .

Abstract ( 1206 )

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Abstract: 【Objective】This study used the DNDC model which is based on the ecological process to simulate the flux of Greenhouse Gas emits from rice paddy field under various management practice scenario and estimate quantitatively. Give one way in accessing the impact of agricultural ecosystem on climate and environment. 【Method】The Landsat TM and CBRES remote sensing images required in 1988 and 2005 were utilized to classify the Land Use/Cover covers Dawa county, Liaoning province. The aim is to extract the temporal-spatial distribution of rice paddy field and calculate its area. The DNDC model was applied to simulate the flux of Gas (Carbon dioxide, Nitrous oxide, Methane) emits from rice paddy field in different period of time, and analyzed the spatial-temporal dynamics of Greenhouse Gas emits from rice paddy field in Liaodong coastal region.【Result】Comparison of the area of rice paddy field in 2005 with that of 1988, there were 7,066.2 hectares increment in amount. The total emission of three GHG (CO2、N2O、CH4) in 1988 and 2005 were-12.02 Pg C, 2.57 Tg N, 0.14 Pg C and 7.22 Pg C, 2.69 Tg N, 0.17Pg C, respectively. The spatial distribution of them were, flux rate of CO2 high region in the east and south of study site, low in the north, N2O high in the mid-east and northwest, low in the north, CH4 high in the mid-east and southwest, low in the north and southeast, respectively. For the net Global Warming Potential (GWP), the spatial difference in 2005 were smaller than in 1988.【Conclusion】Socio-economical impact is the key factor of diving area change of rice paddy field. The rice paddy field is one sink of CO2, source of N2O and CH4. In addition to climate factors, different soil type make significant impact on the flux magnitude of Greenhouse Gas emits from rice paddy field, so does the farm management practice scenario.

The Tupu Analysis of Land Use Change and Its Eco-Environmental Effects Research of Area

Scientia Agricultura Sinica. 2007, 40(10): 2259-2266 .

Abstract ( 1546 )

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This paper constructed the Land Use Changes Tupu During the Period of 1990-2001 in urban and suburban area of Taiyuan city taking advantage of the theory of geographic informatic Tupu supported by RS and GIS. Based on the research of Land Use Changes Tupu, the character of land use change in research area was analyzed and the ecological effects was calculated between 1990 and 2001. The result show that the area of farmland、rangeland and water bodies decreased from 1990 to 2001, the increased areas of farmland were the most, while the area of garden plot、unused land、forest land and structure land increased, of which, the decreased area of forest land were the least. The change of Land Use result in the ecosystem service value of Taiyuan increased from 1202.15×106 yuan in 1990 to 1251.49×106 yuan in 2001,the average increase rate was 0.37％.The most Land Use changes category was the transformation from farmland to unused land, the transformation from farmland to structure land was the next. The degeneration of rangeland、farmland and the increase of unused land brought certain negative affection. By the empirical analysis, the way of geographic Tupu was a very effective meaning in analyzing land use change and ecological effects.

Effect of continuous cucumber cropping on soil microbial populationⅡ

Scientia Agricultura Sinica. 2007, 40(10): 2267-2273 .

Abstract ( 1623 )

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To gain insight into the nature of disease incidences and microbial community shift, the dynamics of soil microbe population induced by continuous cucumber cropping was studied based on PCR-DGGE approach. Results revealed that continuous mono-cropping caused great bacterial species shift, of which Bacteriovorax sp.(indentity of 93%), Pseudomonas sp. (indentity of 97%) and two uncultured bacterial reduced in population, while Sphingomonas sp. (100% similarity) and another uncultured bacterium increased. Monocropping has little effect on eukaryote in soil. For all samples analyzed using culture-independent approach, the bacteria and eukaryotic species shift was more sensitive in rhizosphere than in bulk soils. In the whole growth cycle, the variation of rhizosphere bacterial number was closed coincident with cucumber developmental stages, indicating vigorous root growth and/or root exudates were the potential factors that determined soil bacterial population.

Study of Relativity of Soil Microbial Diversity and Cucumber Yield, Quality in Protected Cultivation

Scientia Agricultura Sinica. 2007, 40(10): 2274-2280 .

Abstract ( 1435 )

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Abstract: 【Objective】 To study the effect of cucumber planting years and cultivated modes on DNA sequence diversity of microbial community and relativity with cucumber yield and quality. 【Method】RAPD techniques were adopted to study the effect of cucumber planting years and cultivated modes on DNA sequence diversity of microbial community and relativity with cucumber yield and quality.【Result】 The results showed that the diversity index, richness and evenness index of microbial community which was in soil of continuous cropping cucumber for 2 years and 7 years decreased with increasing of planting years, and cucumber yield dropped evidently. However, in soil of crop rotation for 15, 18 and 21 years, though the planting years were longer, DNA sequence diversity and evenness of microbial community were both higher than those in soil of continuous cropping cucumber for 7 years. It showed that the rotation of crops is propitious to DNA sequence diversity and stability of microbial community, and also the vitamin C content of cucumber of rotation was higher than that of continuous cropping soil cultivation. DNA sequence diversity index of soil microbial community was positively correlated with cucumber yield and quality evidently.

Preliminary Proteomics Analysis of the Total Proteins of Flower-bud Induction of Apple Trees

Scientia Agricultura Sinica. 2007, 40(10): 2281-2288 .

Abstract ( 1345 )

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Flower-bud formation specific protein of mature spur from apple trees (Malus domestica Borkh.) was studied by employing two-dimensional electrophoresis (2-DE), bioinformatics , and mass spectrometry (MS). The result showed that there were 283 differential protein spots existed and changed in quality and quantity on two 2-DE maps for ‘ Fuji’ at seventh week, of which 162 spots increased and 110 spots decreased in quantity. Four protein spots(16.4、30.2、40.3、65.1kD)were observed the flower bud initial stage 1(inflorescence appears)in two-dimensiononal electrophoresis maps; Three protein spots(39.3、60.2、66.3kD)were observed the initial stage 2(Lateral bud appears) in two-dimensiononal electrophoresis maps; one protein spot(77.1kD)was observed the sepal in two-dimensiononal electrophoresis maps.Four special proteins were observed by peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry. By the analysis of four special proteins spots with peptide mass fingerprinting and matrix-assisted laser desorption ionization time of flight mass spectrometry. When Red Fuji apple begin to spur flower buds. At the same time identified proteins and their putative functions were presented through the retrieval different database. One of them, spot No.256(16.4kD)、298（30.2 kD）were unknown proteins and spot No.327(40.3kD)was identified as the synthesis enzyme protein. Spot No.367(40.3kD)was identified as a RNA-binding protein involved in transcription. Technical details concerning database queries and successful protein identification rate in the absence of a sequenced genome are reported and discussed.

Study on Main Chemical Component of Spongy Mesophyll

Scientia Agricultura Sinica. 2007, 40(10): 2289-2295 .

Abstract ( 1329 )

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The palisade mesophyll and spongy mesophyll of cutter different riped leaf-position (lower,4th,cutter,10th,upper,16th) of Flue-cured Tobacco(K326 andHongDa) were divided by pectinase, then no-volatility organic acid was detected by gas chromatography, and plastid pigment or glucide or nicotine was analyzed by spectrophotometer. It showed that the content of multiple organic acid and nicotine of spongy mesophyll was higher than that of palisade mesophyll, and other components were on the contrary. No matter the palisade mesophyll or spongy mesophyll,the content of chlorophyllous in upper leaf cells was highest, and the content of no-volatility organic acid in lower leaf cells was highest, and the total content of carotin, glucide,deoxidize sugar in cutter leaf cells was highest, and the content of no-deoxidize sugar in cells hasn’t pertinence among the leaf position. The content of nicotine in cells was positive correlation with leaf position.

Genetic analysis of distorted segregation ratio of mating types in Lentinula edodes

Scientia Agricultura Sinica. 2007, 40(10): 2296-2302 .

Abstract ( 997 )

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This study made 17 strains of Lentinula edodes，including wild strains and cultivated strains which well-popularized around China as materials, statistically analyzed the ratios of spores from different aspects via mating types analysis and OWE-SOJ technique. Results from this study firstly systematically identified skewed expected distribution of mating-type factors segregation in Lentinula edodes spores has commonly statistical meanings in wild and cultivated strains. Genetic analysis to positive and negative parental-recombined fruiting showed that nuclear type of F1 progeny spores among those strains segregated theoretically distribution mainly depended on combined state of parental dikaryons, predominant spores were those with mating type identical to dikaryotic parent, indicating that the genetic basis of segregation distortion of spores is different from that of protoplast monokaryons in which B factor takes predominant responsibility for that phenomenon, it cooperate A factor with B factor to influence the ratio of spores.

N-lauroylethanolamine regulation on membrane integrity in carnation petals during senescence

Scientia Agricultura Sinica. 2007, 40(10): 2303-2308 .

Abstract ( 1458 )

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The present study was conducted to examine the possible involvement of N- lauroylethanolamine(NAE(12:0)) in the regulation of petal-membrane senescence. The petal-membrane senescence is accompanied by a decrease in membrane phospholipids content, membrane fluidity, the index of unsaturation of fatty acids and activities of membrane enzymes, and NAE(12:0) supplementation delayed the decreases in above physiology parameters in the aging petals in Dianthus caryophyllus L. ‘Red Barbara’. Electrolyte leakage, which is a reliable indicator of petal-membrane senescence, was also postponed in NAE(12:0)-treated flowers. The data collectively suggest that the NAE(12:0)-induced delay in petal-membrane senescence involved the regulation on the degradation of phospholipids and changes of the fatty acid composition of microsomal membranes, which resulted in the less ion leakage, more fluidity of membrane and higher activities of membrane enzymes, thus preserving the integrity of the petal-membrane and delaying the initial wilting of cut carnation.

Changes of volatile flavor compounds in dry-cured goose during processing

Scientia Agricultura Sinica. 2007, 40(10): 2309-2315 .

Abstract ( 1138 )

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Headspace solid phase microextraction combined with gas chromatgraphy-mass spectrometry（HS-SPME-GC-MS）was used to identify the kinds and amounts of volatile flavor compounds at the different stages of dry cured goose during processing. 112 compounds were identified which included 43 compounds at the raw goose meat, 63 compounds after cured and 50, 51, 62 compounds after drying 1day, 2days, 3days respectively. During processing, the kinds of aldehydes, acids and heterocyclic compounds increased significantly. The total area of the flavor compounds in dry-cured goose, especially in alkenes, aldehydes and ketones increased dramatically during processing and was 3-times more than that in raw material after drying 3 days. All these compounds could play important roles for the specific flavor of dry-cured goose.

Effects of Soybean Oil and Linseed Oil Supplementation on the Rumen Fermentation and Rumen Microflora of Cattle

Scientia Agricultura Sinica. 2007, 40(10): 2316-2322 .

Abstract ( 1038 )

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Abstract: Three rumen-fistulated hybrid yellow cattle were randomly assigned according to a 3×3 Latin square design to investigate effects of supplementation oil in concentrates on pH, NH3-N, VFA and rumen microflora. The dietary treatments were 65% rough and 35% concentrate-based, containing 0 (control), 4% soybeam oil (soybean oil treatment) and 4% linseed oil (linseed oil treatment) of DMI, respectively. The experiment was conducted for three periods, each period lasting 21 days. The results indicated that ruminal pH value was not significantly affected by supplementing oil (P＞0.05). NH3-N concentrates in control and linseed oil treatment were significantly higher than soybean oil treatment (P<0.05). Compared with control group, acetate, propionate, butyrate and total VFA concentration, total viable bacteria, proteolytic bacteria, cellulytic bacteria and protozoa in soybean oil and linseed oil treatments were dramatically lower than the control group. However, there were not significant differences on acetate to propionate ratio and amylolytic bacteria by supplementing oil.

Developmental changes and effect on IMF content of LPL mRNA expression in sheep muscle

Scientia Agricultura Sinica. 2007, 40(10): 2323-2330 .

Abstract ( 1230 )

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Male Kazak sheep and Xinjiang fine wool sheep from 2 to 120-day-old were selected to investigate the developmental changes and effect on Intramuscular fat (IMF) content of lipoprotein lipase (LPL) mRNA expression in sheep muscle. Six sheep of each breed (only Xinjiang fine wool sheep at 120-day-old) were slaughtered at 2, 30, 60, 90, 120 days respectively to sample longissimus dorsi muscle to analyze the IMF content. The total RNA of the samples was extracted to determine the developmental changes of LPL mRNA expression levels by Real time PCR .The results showed that: (1) In male Kazak sheep, the IMF content increased continuously with growing and showed significant difference (P<0.05) between ages, but no difference between ages existed in Xinjiang fine wool sheep (P<0.05). Furthermore, the IMF content of male Kazak sheep was extremely higher (P<0.01) than that of Xinjiang fine wool sheep from 30 to 90 day. (2) LPL mRNA expression level was the highest at 2 day and showed extremely higher than other ages (P<0.01) in male Kazak sheep, it was the lowest at 60 day and then increased. In Xinjiang fine wool sheep, the LPL mRNA was the highest at 30 day (P<0.01), then decreased continuously and to the lowest lever at 90 day; then, the LPL mRNA increased once more. (3) In male Kazak sheep, from 2to 60 day, LPL mRNA expression level was negatively related to IMF content (γ = -0.625, P<0.05), and there was no obviously relationship between LPL and the male Xinjiang fine wool sheep.

Expression Diversity of Calpastatin Gene among Chicken Different Breeds and Tissues

Scientia Agricultura Sinica. 2007, 40(10): 2331-2335 .

Abstract ( 1103 )

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Calpastatin（CAST） gene expression diversity among chicken various breeds and tissues was investigated by using semiquantitative RT-PCR.To collect the tissue samples ,20chickens of 5 breeds were slaughtered.The results indicated that CAST gene were expressed in chicken various kinds tissue. The expression in chest muscle was more significant than in other organiz (P < 0.05).moreover,it was identified that CAST gene expression in Avian Broilers was higher than in Vans Nera egg chicken and adequate quality broilers muscles(P <0.05).

Molecular Cloning and Ontogenesis Expression of Fatty Acid Translocase cDNA in Yellow Feather Broiler

Scientia Agricultura Sinica. 2007, 40(10): 2336-2342 .

Abstract ( 1378 )

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Abstract: 【Objective】To clone chicken FAT/CD36 complete cDNA sequence and study the ontogenesis expression pattern of FAT/CD36 mRNA;【Method】Chicken FAT/CD36 complete cDNA was cloned by 5’- and 3’-RACE. Subcutaneous fat and visceral fat samples were collected and weighed from both male and female Yellow-feathered broilers on d22, d29, d42 and d56 (n=10). Fat content of breast muscle and thigh muscle were determined on d29 and d56. Real time RT-PCR was conducted to investigate the tissue specific ontogenesis expression pattern of FAT/CD36 mRNA in breast muscle, thigh muscle, subcutaneous fat and visceral fat samples collected from both male and female Yellow-feathered broilers;【Result】We have cloned chicken FAT/CD36 complete cDNA sequence of 2243bp(GenBank: DQ323177), which including the 1416bp ORF. The accumulation of subcutaneous fat and visceral fat gradually increased associated with age and the FAT/CD36 mRNA expression levels in subcutaneous fat and visceral fat of male broilers also increased gradually, the highest expression level proved to be in the visceral fat. The FAT/CD36 mRNA expression levels of female broilers were relatively high during the development prophase (d22 and d29), whereas a degressive expression pattern was showed during the development anaphase (d42 and d56);【Conclusion】We have successfully cloned chicken FAT/CD36 complete cDNA sequence. The expression pattern of FAT/CD36 mRNA in muscle and fat were different with gender.

Development of Infectious Molecular Clones of Infectious Bursal Disease Virus Attenuated strain

Scientia Agricultura Sinica. 2007, 40(10): 2343-2349 .

Abstract ( 1476 )

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Abstract: Infectious bursal disease (IBD) ,mediatied by infectious bursal disease virus (IBDV),is one of the causive agents which cause significant losses to the poultry industry. In this research, EcoRⅤ site or PstⅠsite ,as the genetic tags,was introduced separately into the A segment or B segment of the IBDV Gt strain.The full-length genome was flanked by hammerhead ribozyme(HamRz) and hepatitis delta ribozyme(HdvRz) sequence. The full-length genome containing genetic tags flanked by HamRz and HdvRz were arranged downstream of the Beta chincken actin promoter of the vector pCAGG.The recombinant plasmid ，which were infectious clones,were named pCAGGmGtAHRT and pCAGGmGtBHRT.These clones can infection DFⅠcell effectively.The results of RT-PCR,indirect immunofluorescence,electron microscope showed IBDV was rescued successfully. The genome of rescued IBDV have genetic tags.The rescued IBDV could cause CPE on CEF,and the TCID50 of rescued IBDV was similar to Gt.The method of rescuing verus was the basis for further researching IBDV.

The Effect of Monochromatic Light on Broiler Growth

Scientia Agricultura Sinica. 2007, 40(10): 2350-2354 .

Abstract ( 1467 )

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【Objective】 Using LED (light-emitting diodes) as light sources, our study was to compare AA (Arbor Acres) development and productive performance with the monochromatic light and discuss the internal mechanism. 【Method】 Day-old 264 male broilers were divided into four light treatments (n=60), control white (400 nm-700 nm), red (660 nm), green (560 nm) and blue (480 nm) by using LED. There were 3 replications for each light treatment. Feed and water were provided for ad libitum consumption. Light intensity was 15 lux at the height of birds’ heads and scheduled for 23h of light and 1 h of dark during the entire experiment. 【Results】 The results showed that body weight in both groups of blue and green light were higher than that of red and white light groups (6.86 %-10.75%, P<0.05) during the early stage (0-26 day of age) all along. Meanwhile the feed conversion ratio of green light group (1.8) was the lowest in all groups. Body weight of blue light group was the highest during the late stage (27-49 day of age). At the 49 day of age, body weight of blue light group was higher than other groups (17.87 %, P<0.05). Carcass, breast muscle, thigh, crus and net chamber of blue light group were higher than others (14.39%-19.86%, P<0.05) and feed conversion ratio (2.12) was the lowest. Using radioimmunoassay to determine serum testosterone concentration, we found testosterone level was increased following by age during the early stage and it reached the peak at 21 day of age; however, it was decreased slowly in various groups during the late stage. But testosterone level of green and blue light groups was higher than that of red and white group (35.37%-37.13%, P<0.05) during the early stage, but that of blue light group was higher than others during the late stage (14.36%-28.77%, P<0.05). 【Conclusion】This study indicates broiler’s growth and productive performance would be increased when who was illuminated with green or blue light at the early stage and blue light at the late stage under 15 lux light intensity. This concerns that blue and green light were more effective to stimulate to secrete testosterone.

Study the expression map of salt-tolerance mutant under salt-stress in wheat using gene microarray

Scientia Agricultura Sinica. 2007, 40(10): 2355-2360 .

Abstract ( 1046 )

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【OBJECTIVE】To study the reaction of different gene of root under salt-tolerance in wheat;【METHOD】Analysed the gene expression of root in wheat using gene chip technology; 【RESULTS】 The expression of all gene in root of wheat salt-tolerance mutant RH8706-49 were analysed using gene chip technology. The responses to different time of salt stress were studied. The differentia expression map of 61,215 genes was gained. The clustering analysis of hybridizing data showed gene expression changed in root under salt stress. Both salt-induced gene and salt-inhabited gene had been found;【CONCLUSION】The mechanism of salt-tolerance in wheat is very complicated. It is the result of a lot of genes expression in line. And the function of the salt-induced gene is very important.

Character of the hormones from the calli of rice

Scientia Agricultura Sinica. 2007, 40(10): 2361-2367 .

Abstract ( 1132 )

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We have detected the endogenous hormone of the calli from four main processes of rice’s genetic transforming.The results shows endogenous hormone distributing nonuniformity in different part of calli training materials and the growth potential are relevance to the content of some endogenous hormone, and the endogenous hormone are changed intensity at different phase of the rice’s genetic transforming

Studies on stigma receptivity and pollen viability in Zinnia elegans

YaoMei Ye

Scientia Agricultura Sinica. 2007, 40(10): 2376-2381 .

Abstract ( 1622 )

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In order to improve the effect of hand-pollination in hybridization of Zinnia elegans the stigma receptivity and pollen longevity were tested. The studies were put in practice connecting the benzidine-H2O2 testing stigma receptivity , pollen germination test in vitro and the experiment of hand-pollination. The results show that Zinnia elegans flowers at 8:30-16:00 in daytime, pollen viability is for 18h. When it flowers, the stigmas of florets stand upright and keep high turgidity for 7~10 days. The benzidine-H2O2 test shows that the Stigmatic Pollen Receptivity for 1~3d, 4~6d and 10d are 79.9%~83.1%, 56.1%~63.0% and 56.1%~63.0% respectively, and there is an obvious difference for the stigmatic pollen receptivity between 1~6 d and 7~10 d. The experiment of hand-pollination shows that the seed set of the丫 stigma phase, the tip of 丫 slightly curling stigma phase and columnar stigma phase is 76.3%、48.5%,36.8% respectively. The seed set of no-treatment, detruncating one lob of stigma and style of detruncating stigma is 48.8%,30.9%,19.9% respectively, this indicates the stigma top with hair and its big surface is in favor of seeding, and detruncating stigma by hand-pollination also may seed. We should pollinate every other day at 10:00-16:00 daytime during flowering season to insure good seed set.

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