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Table of Content

    10 October 2009, Volume 42 Issue 10
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Analysis of Genetic Similarity of Improved Japonica Rice Varieties from Different Provinces in China
    SHU Ai-ping ,ZHANG Yuan-yuan,CAO Gui-lan,LU Qin,ZHANG San-yuan,HAN Long-zhi
    Scientia Agricultura Sinica. 2009, 42(10):  3381-3387 .  doi:10.3864/j.issn.0578-1752.2009.10.001
    Abstract ( 1191 )   PDF (234KB) ( 1168 )   Save
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    【Objective】 The genetic similarity and genetic difference among improved japonica rice varieties from different provinces of China were analyzed. The aim is to provide genetic basis for the breeding of japonica rice varieties. 【Method】 The genetic similarity and cluster of 139 accessions of improved japonica rice variety from 12 provinces of China were analyzed using 34 SSR markers. 【Result】 Totally 198 alleles were detected among these improved japonica rice varieties with the average number of alleles per pair of primers was 5.3235. RM320, RM531, RM1, RM286 and RM336 showed more alleles, which were 15, 12, 11, 9 and 9, respectively. RM320, RM336, RM286 and RM531 showed higher genetic diversity indexes, which were 2.3324, 2.0292, 1.8996 and 1.7820, respectively. The range of genetic similar index among improved japonica rice varieties from different provinces was from 0.321 to 0.914, with the average of 0.686. There is a high genetic similarity among improved japonica rice varieties from Heilongjiang, Jilin, Liaoning, Ningxia, and Yunnan, which were located in similar latitude or similar ecological environment, while there is a low genetic similarity between improved japonica rice germplasm from Guizhou and Jiangsu, and those of other provinces, which located in more different latitudes and ecological environments. 【Conclusion】 The marker of RM320, RM531, RM1, RM286, and RM336 fit to be used in analysis of genetic diversity for improved japonica rice variety. The genetic similarity among improved japonica rice varieties from different provinces was closely associated with genetic basis of parents, and was also correlated with latitude and ecological environment where the varieties were bred.

    Analysis of Indica-japonica Differentiation in Rice Parents and Derived Lines Using ILP Markers
    XU Xu-ming,LIANG Kang-jing,ZHANG Shou-gang,SHANG Wei,ZHANG Ying-ying,WEI Xin-yu,KE Bei
    Scientia Agricultura Sinica. 2009, 42(10):  3388-3396 .  doi:10.3864/j.issn.0578-1752.2009.10.002
    Abstract ( 1279 )   PDF (440KB) ( 990 )   Save
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    【Objective】 Revealing the Indica-Japonica differentiation in parents of hybridization between Indica and Japonica rice and their derived lines can provide theoretical and practical bases for the breeding of practical inter-subspecific hybrid rice. 【Method】Using subspeeies-speeific molecular markers ILP (Intron Length Polylnorphism) and Cheng's Index, the indica-japonica differentiation was analyzed with special materials including 18 indica-japonica hybrid parents and 39 derived lines, which accumulated different wide compatibility and restoring genes by convergent cross method in twenty-one years spanning four breeding phases. 【Result】 The indica-japonica differentiation was detected on all tested loci in 57 materials. Among the 18 parental lines, four were japonica type, five japonicaclinous type, eight indicaclinous type and one indica type. The japonica proportion index in indica restorer lines Minghui63 and 9308 were 12.50% and 33.33%, respectively, while that in japonica restorer line C418 was only 31.25%. Among the 39 derived lines from indica–japonica hybridization, one was Japonica type, eleven japonicaclinous type, twenty indicaclinous type and seven indica type. The japonica proportion index in Minghui502 was only 10.42%. The results of indica and japonica classification by ILP molecular marker and Cheng's Index were relatively consistent. The correlation coefficient between the japonica proportion index and morphology index was 0.794**, while that between the indica proportion index and morphology index was -0.7662**.【Conclusion】 ILP marker could be used to accurately detect the proportion of indica/japonica content in the genome of a rice variety. The results of indica-japonica differentiation analysis could make reasonable explanation for that the hybrids obtained from indica-japonica type restorer lines had obvious heterosis. This conclusion would provide important guidance in efficient use of beneficial genes of inter-subspecific hybrid rice.

    Genetic Diversity of the Founder Parent Orofen and Its Progenies Revealed by SSR Markers
    LI Xiao-jun,XU Xin,LIU Wei-hua,LI Xiu-quan,LI Li-hui
    Scientia Agricultura Sinica. 2009, 42(10):  3397-3404 .  doi:10.3864/j.issn.0578-1752.2009.10.003
    Abstract ( 1173 )   PDF (439KB) ( 1031 )   Save
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    【Objective】 The main objective is to uncover the genetic contribution of founder parent to its offspring through tracking chromosomal regions of Orofen in its progeny. 【Method】 Orofen and its 23 representative progenies were analyzed using morphological traits in five environments and microsatellite markers. 【Result】 The frequencies of the alleles which were amplified in Orofen but absent in other 4 parental accessions (Jinan 4, Beijing 8, 54405 and Heine Hvede) at 43 SSR loci were higher than the theoretical ratio 0.38 in the first generation; the frequencies of the alleles which were amplified in Orofen but absent in other 6 parental accessions (Jinan 4, Beijing 8, 54405, Heine Hvede, Lovrin 10 and Lovrin 18) at 41 SSR loci were higher than the theoretical ratio 0.18 in the second generation; at twenty-one of the found loci above, the unique alleles amplified only in Orofen have been selected continuously in the both generations. Association mapping indicated that the alleles according to Orofen which occurred in most progenies at the loci Xwmc710, Xbarc235 and Xbarc252 were associated with higher grain number and yield when compared with the remaining amplified alleles at the same loci. 【Conclusion】 It was speculated that the genomic region detected and associated traits played an important role for the improvement of wheat in China and could be acted as the emphases for further study.

    Isolation and Analysis of TaGA20ox2 Genes in Wheat
    WU Jing,KONG Xiu-ying,GAO Li-feng,REN Zheng-long,JIA Ji-zeng
    Scientia Agricultura Sinica. 2009, 42(10):  3405-3412 .  doi:10.3864/j.issn.0578-1752.2009.10.004
    Abstract ( 1208 )   PDF (671KB) ( 1281 )   Save
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    【Objective】 The objectives of this study are to isolate full length GA20-oxidase(GA20ox2) genes from hexaploid wheat (Triticum aestivum) and locate GA20ox genes on wheat genomes, give a better understanding of the character and evolutionary relationships of GA20ox genes in wheat. 【Method】 Clone GA20ox genes from hexaploid wheat through homologous cloning method and BAC screening were used. 【Result】 Sequences analysis showed that wheat GA20-oxidase has three sequences:TaGA20ox2-A1, TaGA20ox2-B1, and TaGA20ox2-D1. Sequence alignment proved wheat GA20-oxidase genes have the highest homology with rice GA20-oxidase(80% cDNA sequence identity). A series of Chinese Spring nulli-terasomic stocks were employed to ascertain the chromosomal location of wheat GA20-oxidase gene. The results showed that TaGA20ox2-A1, TaGA20ox2-B1, and TaGA20ox2-D1 were on wheat chromosome 3A, 3B, and 3D. Using a population of recombinant inbred lines from the cross W7984×Opata85, TaGA20ox2-D1 was mapped between SSR markers xfba330 and xgwm664 on chromosome 3D, and the map distance was 4.1cM and 5.3 cM, respectively. 【Conclusion】 The full length of TaGA20ox2 was isolated and located on wheat chromosome 3A, 3B and 3D. TaGA20ox2-D1 was mapped between SSR markers xfba330 and xgwm664 on chromosome 3D. In this region a plant height loci was reported. It is speculated that TaGA20ox2-D1 gene maybe control wheat height development.

    QTL Mapping of Seedling Growth Traits and Grain Yield Under Two Nitrogen Conditions in Maize
    LIU Jian-chao,LI Jian-sheng,MI Guo-hua,CHEN Fan-jun,ZHANG Fu-suo
    Scientia Agricultura Sinica. 2009, 42(10):  3413-3420 .  doi:10.3864/j.issn.0578-1752.2009.10.005
    Abstract ( 988 )   PDF (535KB) ( 973 )   Save
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    【Objective】 The purpose of this experiment was to study the genetic linkage between seedling growth and grain yield of maize under different nitrogen (N) applied. 【Method】 An F8 maize RIL population derived from an elite hybrid maize YY 22 (Z3×87-1) was used to identify the QTL for seedling growth traits and grain yield under high nitrogen (HN) and low nitrogen (LN) conditions. QTL for shoot dry weight (SDW), root dry weight (RDW), the total root length (TRL), root/shoot ratio (R/S) and grain yield (GY) were identified by composite interval mapping. 【Result】 Totally, 22 QTL were detected by using the software Windows QTL Cartographer version 2.5 (LOD>2.5). Ten QTLs were detected under HN condition and 12 QTLs under LN condition. Few loci were detected under both LN and HN conditions for the investigated traits, suggesting there is a different genetic basis controlling maize growth at LN versus HN conditions. The genetic linkages were found between seedling growth traits and grain yield in chromosome 5 and 7. 【Conclusion】 Root traits at seedling stage are very important for grain yield. It could be a critical factor for developing N efficient cultivars in maize.

    Analysis on Genetic Diversity and Genetic Basis of Main Sesame Cultivars Released in China
    SUN Jian,ZHANG Xiu-rong,ZHANG Yan-xin,CHE Zhuo,HUANG Bo
    Scientia Agricultura Sinica. 2009, 42(10):  3421-3431 .  doi:10.3864/j.issn.0578-1752.2009.10.006
    Abstract ( 964 )   PDF (596KB) ( 891 )   Save
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    【Objective】 The aim of the experiment is to investigate the genetic diversity and genetic basis of main sesame cultivars in China. 【Method】 SRAP (Sequence-related amplified polymorphism) was used for the analysis of 67 sesame cultivars widely used in China’s sesame major production areas from 1950 to 2007. 【Result】 A total of 561 bands were amplified using 21 SRAP random primer pairs, with 265 of them were polymorphic, resulting a polymorphism ratio of 47.2%. The total bands and polymorphism amplified by each primer pair averaged 26.7 and 12.6, respectively. The average genetic similarity coefficient and genetic distance of the 67 cultivars were 0.9104 and 0.0706, respectively, indicating limited genetic diversity and narrow genetic basis. Comparative analysis on genetic similarity and genetic distance of different classified cultivars showed that the difference of average genetic similarity coefficient and genetic distance between the landraces and cultivars bred through crosses reached a significant level (P=0.01), with the genetic basis of landraces wider than that bred cultivars. The genetic basis of cultivars used in 1990-2007 was more narrow than that of cultivars from 1950 to 1969 and from 1970 to 1989, with the differences of average genetic similarity coefficient and genetic distance reached a very significant level. 【Conclusion】 The genetic basis of main sesame cultivars in China is relatively narrow, and the genetic basis of cultivars developed through crosses in recent years is more narrow than history cultivars.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
    A Difference in Nitrogen Uptake and Distribution in Conventional Indica Rice Cultivars with Different Sink-Potentials
    DONG Gui-chun,YU Xiao-feng,DONG Yan-ping,LI Jin-qian,TIAN Hao,ZHOU Juan,WANG Yun-xian,YANG Lian-xin,HUANG Jian-ye,WANG Yu-long
    Scientia Agricultura Sinica. 2009, 42(10):  3432-3441 .  doi:10.3864/j.issn.0578-1752.2009.10.007
    Abstract ( 1044 )   PDF (306KB) ( 934 )   Save
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    【Objective】 The goal of this research was to analyze the characteristics of nitrogen uptake and distribution in conventional indica rice cultivars with large sink potential(SP). 【Method】 Eighty-eight and 122 conventional indica rice cultivars were solution-cultured in 2001 and 2002, respectively. Dry matter weight (including root system, culm and sheath, leaves, panicle), nitrogen content in different organs, yield and its components were measured. The tested rice cultivars were classified into 6 types (i.e. A, B, C, D, E and F, A was the lowest, and F was the highest) based on their SP level by the MinSSw method, to study their difference in nitrogen uptake and distribution. 【Result】 Difference of SP of the cultivars used in this study was very large, the averages of SP in different types were 426.37, 642.53,770.96, 903.73, 1064.32, 1213.90 g?m-2 in 2001, and 359.36, 574.11, 764.98, 962.43, 1200.11, 1455.59 g?m-2 in 2002, respectively; in large SP types of indica rice, N content (%) at heading stage was higher, while that decreased more during grain filling period. Cultivars with large SP were characterized by higher N accumulation during grain filling period and at mature stage, lower rate of N distributed in root, leaf, culm, and sheath and higher translocation rate from leaf, culm and sheath to panicle. N accumulation was significantly influenced by growth duration and N uptake intensity, but the effect of N uptake intensity on N accumulation was more important than growth duration. Enhancing N accumulation and rate of nitrogen translocation in culm, sheath and leaf during the grain-filling stage could improve SP. 【Conclusion】 Cultivars with large SP were characterized by higher N accumulation during grain filling period and at mature stage, lower rate of N distributed in vegetative organs and higher translocation rate from leaf, culm and sheath to panicle.

    Effects of the Decreased Index of SPAD Value of Leaf After Full Heading on Ratooning Ability

    XU Fu-xian,XIONG Hong,ZHANG Lin,GUO Xiao-yi,ZHU Yong-chuan,ZHOU Xing-bing,LIU Mao
    Scientia Agricultura Sinica. 2009, 42(10):  3442-3450 .  doi:10.3864/j.issn.0578-1752.2009.10.008
    Abstract ( 939 )   PDF (363KB) ( 672 )   Save
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    【Objective】 Many studies on the source-sink characters of high-yielding combinations of main crop or ratooning rice, but there were few studies on high-yielding combinations total yield of main crop and ratooning rice. In this paper, the relationship between the decreased index of SPAD value of leaf from full heading to harvesting (DISVLFHH) and ratooning ability,the plant characters of main crop for high yielding combinations of main crop and ratooning rice were studied, which could provide a theoretical and practical basis for improvement of high yield breeding and cultivation of hybrid rice. 【Method】 The experiment was conducted with eighteen mid-season rice combinations grown in a traditional high yield cultivation system, in a randomized block design with 3 replications. The results were analyzed through variance, correlation, regression and path analysis. 【Result】Alive bud rate, born bud rate, effective panicles and grain yield of ratooning rice were the key traits of ratooning ability, which was a an important approach for further high yield of ratooning rice by increasing effective panicles.There were significant or highly significant relationships between spikelets per panicle,the ratio of leaf area to spikelets, the ratio of leaf area to grains, the ratio of leaf area to grain weight, decreased LAI index, DISVLFHH of main crop and alive bud rate, born bud rate, effective panicles, grain yield of ratooning rice, and the partial correlation coefficient between only DISVLFHH among all trais and alive bud rate, born bud rate, effective panicles, grain yield of ratooning rice got significant at 0.05 level. As for one variety, it’s impossible that both main crop and ratooning rice could get top grain yield. The high-yielding combinations for 11.5t per ha of the theory grain yield and 14t per ha of potential grain yield for main crop and ratooning rice would have the following source-sink characteristics: 160-190 spikelets per panicle, the ratio of leaf area to grain weight 0.0737-0.0827 cm2 per mg , DISVLFHH 0.4029-0.5409, effective panicles 232.12×104-249.40×104 per ha, seed setting percentage 81.54%-85.74%, 1000-grain weight 28.58-30.07 g, grain weight 4.13-4.43 g per panicle. 【Conclusion】 Increasing effective panicles is an important approach for further high yield of ratooning rice, DISVLFHH of main crop can be used as a new evaluation index for ratooning ability,and the combinations with middle-big spikelets per panicle are the key characteristic for selection of mid-season rice hybrids for high yield of main crop and ratooning rice.

    Effects of Shading From Jointing Stage to Maturity Stage on High Molecular Weight Glutenin Subunits Accumulation and Glutenin Macropolymer Content in Wheat Grain
    BO Yun,LI Hua-wei,MU Hui-rong,CAI Jian,ZHOU Qin,DAI Ting-bo,CAO Wei-xing,JIANG Dong
    Scientia Agricultura Sinica. 2009, 42(10):  3451-3458 .  doi:10.3864/j.issn.0578-1752.2009.10.009
    Abstract ( 1253 )   PDF (620KB) ( 690 )   Save
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    【Objective】 Accumulations of high molecular weight glutenin subunits (HMW-GS) and glutenin macropolymer (GMP) play key roles in determining grain quality in wheat (Triticum aestivum L.). Two wheat cultivars owning the same HMW-GS type (7+8 and 2+12) were selected to study the effects of shading from jointing to maturity on HMW-GS accumulation and GMP content in grain. 【Method】 HMW-GS was firstly separated using the sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE), and each HMW-GS subunit band in the gel was cut and extracted. Content of each HMW-GS subunit was then quantified by colorimetric analysis of the HMW-GS subunit extraction. 【Result】 Shading from jointing and maturity promoted the initial formation time of HMW-GS. The severe shading treatments (18% and 25%) shortened the rapid accumulation period of HMW-GS and reduced the accumulating rate of total HMW-GS during late grain filling period. Thus, the accumulation amount of total HMW-GS at maturity was lower under these treatments than the non-shading treatment. However, contents of HMW-GS and GMP were higher under severe shading than the control. A low-degree shading treatment (10%) prolonged the rapid accumulation period of HMW-GS, and increased the final accumulation amount and content of total HMW-GSP. 【Conclusion】 Shading from jointing to maturity obviously affected the accumulation amounts and contents of total HMW-GS in wheat grain. However, this regulatory effect was related to shading intensity.

    Effects of Irrigation and Urea Types on Ear Leaf Senescence After Anthesis, Yield and Economic Benefit of Maize
    SHAO Guo-qing,LI Zeng-jia,NING Tang-yuan,JIANG Bao-juan,JIAO Nian-yuan
    Scientia Agricultura Sinica. 2009, 42(10):  3459-3466 .  doi:10.3864/j.issn.0578-1752.2009.10.0010
    Abstract ( 949 )   PDF (298KB) ( 964 )   Save
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    【Objective】 The objective of the study is to investigate the effects of application rates of normal urea (NU) and controlled release urea (CU) on ear leaf senescence, yield and economic benefit of summer maize under different water conditions. 【Method】 Five nitrogen rates and two irrigation levels were designed to investigate the effects of water-nitrogen coupling on photosynthetic rate, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), malondialdehyde (MDA) content and soluble protein content in maize ear leaves after anthesis, and their relationship with grain yield and economic benefit. 【Result】 At the same irrigation level, net photosynthetic rate of maize applied with controlled CU was lower (P < 0.05) than that applied with NU at anthesis, but the situation was just contrary after anthesis. At the same nitrogen level, compared with the NU, CU can significantly increase water content, activities of SOD, POD and CAT, and the content of soluble protein in ear leaves, but reduce MDA content. So, the grain yield and economic benefit of maize applied with CU was higher than that applied with NU, and irrigation is beneficial to improve the superiority of CU. But because of the higher cost of CU, when higher CU were used with irrigation, the economic benefit was lower than NU. 【Conclusion】 Compared with the NU, the positive coupling effect of CU might be caused by the higher SOD, POD and CAT activities and soluble protein content, and the lower MDA content after anthesis, which could delay the senescence and increase the net photosynthetic rate of ear leaf, and as a result, increase the grain yield of maize. With the increase of CU production, its cost will be decreased and its use prospect will be wider in crop production.

    Effects of Interspecific Interactions and Nitrogen Fertilization Rates on the Agoronmic and Nodulation Characteristics of Intercropped Faba Bean
    LI Yu-ying,SUN Jian-hao,LI Chun-jie,LI Long,CHENG Xu,ZHANG Fu-suo
    Scientia Agricultura Sinica. 2009, 42(10):  3467-3474 .  doi:10.3864/j.issn.0578-1752.2009.10.0011
    Abstract ( 1025 )   PDF (354KB) ( 1005 )   Save
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    【Objective】 The objective of this paper is to investigate the dynamics of the agronomic traits and nodulation of faba bean intercropped with maize as affected by intercropping and N application rates, therefore to provide a scientific basis for the N nutrient management with maximizing grain yield and nutrient efficiency of legume/cereal intercropping system in Hexi Corridor of Gansu Province. 【Method】 The N gradient experiment and micro-plot root barrier experiment were conducted using split-plot design and row root barrier technique at Baiyun Experimental Station of Gansu Academy of Agricultural Sciences during 2006-2007, so as to study the effects of interspecific interactions and N application rates on the agronomic traits and nodulation of intercropped faba bean. 【Result】 N application rates had no significant effect on the above-ground growth and the parameters of yield traits of faba bean. However, the nodulation of faba bean was significantly inhibited by N fertilization and the inhibitory effect was increased with N fertilization rate increasing and faba bean growing. Interspecific facilitation on the nodulation of faba bean was increased with co-growth of both crops. Intercropping significantly enhanced the nodulation size of faba bean and alleviated “Nmin inhibitory effect” on nodulation, which the value was 14.4%. The nodulation size of faba bean with interspecific root interactions increased by 10.2%, compared to that without interspecific root interactions. 【Conclusion】 The interspecific root interactions between faba bean and maize significantly enhanced the nodulation and alleviated “the Nmin inhibitory effect”, and significantly increased the yield of intercropped faba bean.

    Effects of Light Intensity and Fertilization on the Growth of Poa crymophila

    MA Yin-shan,WANG Xiao-fen,ZHANG Zuo-liang,LI Chuan-long,DU Guo-zhen,ZHANG Shi-ting
    Scientia Agricultura Sinica. 2009, 42(10):  3475-3484 .  doi:10.3864/j.issn.0578-1752.2009.10.0012
    Abstract ( 914 )   PDF (352KB) ( 709 )   Save
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    【Objective】 This study is to examine the effects of light intensity and fertilization on the growth of Poa crymophila in Alpine meadow and to provide insights into the proper utilization and the optimal management of the meadow. 【Method】 Through a factorial design in field, the experiment included eighteen 1.3 × 0.8 m plots with a total of 6 treatment combinations of light and fertilization. The intensity of light was divided into three levels: 100% of light (strong), 43.5% of light (medium) and 6.74% of light (weak). The treatment of fertilization included two levels: fertilized and non-fertilized. The parameters of plant growth (tiller numbers, plant height, above-ground biomass, specific leaf area, relative growth rate and the resource allocations to different plant parts) were measured to assess the adaptation of this herb to dissimilar circumstances. 【Result】 The results showed that the number of tiller and above-ground biomass of P. crymophila decreased as the light reduced, which could be compensated by fertilization. The plant reached its maximum of height under the medium intensity of light. The fertilization promoted the growth of plant under the strong and weak intensity of light. When the light weakened, the SLA increased but the RGR reduced, while fertilization had no effects on them at all. Root allocation and root/shoot ratio had no significant difference between treatments under medium and weak intensity of light respectively, but the fertilization could lead to the decrease of root allocation and root/shoot ratio under the strong light. When the intensity of light reduced, the leaf allocation increased while the reproduction allocation reduced, while fertilization had no influence on the leaf and the reproduction allocation. The stem allocation decreased with the reduced intensity of light under the fertilization treatment. Under the non-fertilized treatment, the stem allocation was the largest under the medium light treatment rather than the strong and the weak light intensity. 【Conclusion】 P. crymophila is sensitive to light as the decrease of light has an negative effect on its growth. The strong light and the fertilization can benefit the plant growth, while the medium light can weaken the effect of fertilization, and the negative effect of weak light can be compensated by the fertilization treatment. For the grassland under canopy dominated by P. crymophila, the fertilization is ineffective when the light intensity permeated through the gap of canopy can reach the medium level; however, if it achieves the low light level, the grass production can be increased by fertilization.

    PLANT PROTECTION
    Efficacy and Mechanism of Control of Wheat Stripe Rust by Diversifying Cultivars in Mix-Planting
    GUO Shi-bao,HUANG Li-li,KANG Zhen-sheng,CHENG Jing-jing,LU Ning-hai,YANG Zhi-wei,CHEN Yin-chao
    Scientia Agricultura Sinica. 2009, 42(10):  3485-3492 .  doi:10.3864/j.issn.0578-1752.2009.10.0013
    Abstract ( 1242 )   PDF (522KB) ( 960 )   Save
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    【Objective】 To provide a theoretical basis for the use of mix-planting as an ecological control method for wheat stripe rust, the control efficacy and mechanism of wheat stripe rust by diversifying cultivars in mix-planting were studied. 【Method】 Six mono-cultivars treatments and nine mix-cultivars treatments were designed and disease incidence, foci numbers and yield per plot were investigated. Furthmore, TP-M13-SSR technique was employed to detect population genetic diversity of P. striiformis f. sp. tritici in natural infested wheat in mono-cultivar and mix-cultivar plots. 【Result】 The foci numbers and development of disease at seedling stage were obviously inhibited on mix-cultivar plots compared to mono-cultivar plots. The disease index was significantly decreased during spring epidemical stage on mix-cultivar plots. The mixture planting of resistant and susceptible cultivars showed better efficacy in disease control and more reduction of yield losses. The relative control efficacy was 73.27% and yield increase rate was 13.26%, on the average, compared to mono-cultivar planting. Cultivar numbers in mix-cultivar planting did not showed the obvious effect on disease control and yield production. Population genetic diversity of P. striiformis f. sp. tritici was higher in mix-planting than that in mono-planting. 【Conclusion】 The mix-cultivar planting showed higher control effect on wheat stripe rust and more reduction of yield losses than mono-cultivar planting. The mix-cultivar planting could be used as one of the ecological measures to prevent and control disease.

    Detection of Tilletia controversa with HRCA Approach
    CAI Jun,YIN You-ping,GE Jian-jun,CHEN Hong-jun,HUANG Guan-jun,ZHANG Wen-di,WANG Zhong-kang
    Scientia Agricultura Sinica. 2009, 42(10):  3493-3500 .  doi:10.3864/j.issn.0578-1752.2009.10.0014
    Abstract ( 1145 )   PDF (704KB) ( 1385 )   Save
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    【Objective】 To establish a detection system of Tilletia controversa (TCK) with hyper-branched rolling cycle amplification (HRCA) method, which provides a stable , reliable and novel technique for early diagnosis of wheat dwarf bunt disease and identification of pathogen. 【Method】 The padlock probe consists of a universal linking sequence and the two target complementary regions at 5′and 3′ends, which was designed based on the unique fragment sequence of 1 322 bp of TCK. Detection system of HRCA was established and optimized. The specificity and limitation of HRCA was determined and compared with conventional PCR. A total of 51 samples intercepted at Customs or collected from different wheat cultivation areas in USA and China were examined with HRCA. 【Result】 HRCA is capable of amplifying mycelial and teliosporal DNA of TCK , while not detecting related smut species of Tilletia and other plant pathogen. The detection sensitivity of HRCA is as low as 1 fg?μl-1 for plasmid DNA and 10 pg?μl-1 for genomic DNA of TCK which is 10-fold higher than that of conventional PCR. The results show HRCA method to be sensitive, specific and accurate. 【Conclusion】 The detection system of HRCA for TCK was successfully established, which provided a new approach for the simultaneous diagnosis of wheat dwarf bunt disease and identification of TCK pathogen.

    DNA Microarray Expression Analysis of Xanthomonas oryzae pv. oryzae in Rice Leaves at Early Infection Stages Using Selective Bacterial Transcript Labeling with Genome-Directed Primers
    ZHANG Xin-jian,GAO Shi-qiang,WU Mao-sen,HE Chen-yang
    Scientia Agricultura Sinica. 2009, 42(10):  3501-3508 .  doi:10.3864/j.issn.0578-1752.2009.10.0015
    Abstract ( 1017 )   PDF (267KB) ( 664 )   Save
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    【Objective】 The objective of this study is to reveal the bacterial gene expression profiling of Xanthomonas oryzae pv. oryzae (Xoo) in rice leaves at early infection stages via DNA expression microarray analysis. 【Method】 A computer-based algorithm for prediction of the minimal number of primers to specifically anneal to all genes in the genome (genome-directed primers, GDPs) of KACC10331, a genome sequenced strain of Xoo was used. Seventy-four oligonucleotides (8 mer in length each) priming all genes in the genome were predicted and used. A set of microarray of 371 pathogenicity-related genes was hybridized with the GDPs-primed and fluorescent Cy3/Cy5-labeled cDNAs, which were reverse-transcribed from RNAs isolated from rice leaves 1d, 3d and 7d post-inoculation of Xoo. 【Result】 The differential expression profiling of pathogenicity-related genes of Xoo at different early infection stages was found. Forty-two, fifty-one and thirty-three genes of Xoo were differentially expressed in the rice leaf tissues at 1 d, 3 d and 7 d after bacterial inoculation relative to the nutrition rich media NBY, respectively. Among them, 5 up-regulated genes and 5 down-regulated genes were observed in Xoo at all early infection stages. The expression of Xoo genes might be related to bacterial invasion, adaptation, proliferation and appearance of symptom. 【Conclusion】 The approach of selective transcript labeling with GDPs for in planta gene expression profiling of Xoo is reported for the first time. The differentially-expressed genes of Xoo might be the candidate targets for the further functional analysis.

    Effect of chitinases produced by Pochonia chlamydosporia on egg-hatching of Meloidogyne incognita
    ZHANG Cheng-min,WU Xia,CAI Xiu-hua
    Scientia Agricultura Sinica. 2009, 42(10):  3509-3515 .  doi:10.3864/j.issn.0578-1752.2009.10.0016
    Abstract ( 1211 )   PDF (425KB) ( 1213 )   Save
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    【Objective】 Chitinases produced by egg-parasitic fungus Pochonia chlamydosporia have been suggested to play an important role in digestion of root knot and cyst nematode eggshells. The egg-parasitic fungi with chitinase activity is one of the most important biochemical characters in suppression of egg hatching, and can be used for evaluation of the biocontrol potential of those fungi. 【Method】 The chitinolytic systems and exochitinase produced by 7 isolates of P. chlamydosporia were measured by means of NAG(N-acetyglucosamine) and pNP(ρ-nitrophenol) respectively. 【Result】 The results showed that all the isolates hade different levels of exochitinase activity, however only the isolates QNAV97-2, NRRL13094, CFCC84964 and CFCC80919 had activity of chitinolytic systems. On SDS-PAGE analysis with 0.01% glycol chitin, two and four protein bands with chitinase activity were detected, the chitinases of 38.9kDa and 39.8kDa produced by CFCC80964 and QNAV97-2 were believed to be the CHI43 purified in early studies. Inhibitory rates to egg hatching of Meloidogyne incognita by 4 isolates with chitinolytic activity were ranged from 40.32% to 55.15%, however, those by 3 isolates without chitinolytic activity were lower than 20%. Microscopy observations demonstrated that the eggshells of M. incognita was deformed and destroyed in the treatment of culture filtrates from P. chlamydosporia. 【Conclusion】 It is concluded that the chitinases produced by P. chlamydosporia causes lysis of the eggshell of M. incognita, especially immature eggs at early embryonic development stage, and result in egg hatching inhibition and/or egg kill. The digestion of the eggshell by chitinase activity plays an important role in the control of M. incognita. Difference in chitinase activity levels produced by P. chlamydosporia may result in their difference in nematicidal activity to nematodes.

    Molecular Basis of Resistance to Bensulfuron-Methyl in Monochoria korsakowii
    LU Zong-zhi,ZHANG Chao-xian,FU Jun-fan,LI Mao-hai,LI Gui-jun
    Scientia Agricultura Sinica. 2009, 42(10):  3516-3521 .  doi:10.3864/j.issn.0578-1752.2009.10.0017
    Abstract ( 1030 )   PDF (418KB) ( 822 )   Save
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    【Objective】 In recent years, Monochoria korsakowii has been a serious problem in paddy fields in northeast of China. It has become a weed which is very difficult to control by bensulfuron-methyl in some areas in the region. The objective of this study is to understand the molecular basis of the resistance mechanism to bensulfuron-methyl in Monochoria korsakowii and to find the specific mutation sites in amino acid sequence of acetolactate synthase (ALS) in the resistant Monochoria korsakowii biotype. 【Method】Using the PCR method, fragments that encode the ALS were cloned from Monochoria korsakowii, susceptible (S) and resistant (R) biotypes to bensulfuron-methyl respectively, and sequenced subsequently. 【Result】 The result showed that the nucleotide sequence of R M. korsakowii biotype differed from that of the S biotype by three nucleotide substitutions, that were proline (Pro) 197 substituted by histidine (His), methionine (Met) 200 by valine (Val), and arginine (Arg) 388 by histidine (His),among those, the amino acid substitution of Pro197 (CCT) to His (CAT) had been reported in many other resistant weeds. 【Conclusion】 It is clear that the substitution of Pro197 may be responsible for the resistance to bensulfuron-methyl in the R M. korsakowii biotype. The functions of the other two mutations in the R biotype need to be further investigated.

    Effect of New Herbicide ZJ0273 on Seedling Growth and Root Cell Viability of Brassica napus
    ZHANG Fan,TIAN Tian,JIN Zong-lai,HUANG Chong-ping,TANG Gui-xiang,YE Qing-fu,ZHOU Wei-jun
    Scientia Agricultura Sinica. 2009, 42(10):  3522-3529 .  doi:10.3864/j.issn.0578-1752.2009.10.0018
    Abstract ( 1170 )   PDF (398KB) ( 939 )   Save
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    【Objective】This paper described the response of seedling growth and root cell viability of Brassica napus L. (rapeseed) to various concentrations of ZJ0273 treatments at germination stage to define the mechanism of this novel herbicide.【Method】Effects of different ZJ0273 treatments on dry matter, morphological characters, root oxidizability and cell membrane permeability of rapeseed seedlings were investigated by various physiological measurements. Root cell viability and mitosis as affected by herbicide treatments were also studied based on FDA-PI double staining and root-tip squashing method. 【Result】The results indicated that the inhibitive effects of ZJ0273 treatments on rapeseed dry matter and morphological characters were enhanced along with the increase of treatment concentrations and durations. Both of 10 and 100 mg?L-1 ZJ0273 treatments inhibited root development significantly. There were no distinct difference among 0, 0.1 and 1 mg?L-1 ZJ0273 treatments on root oxidizability and membrane permeability, compared with treatments at 10 and 100 mg?L-1 which obviously inhibited root cell viability and induced cell membrane disintegration. Furthermore, mitotic index of root-tip cells was declined and cell division was stopped at metaphase after being treated by 100 mg?L-1 ZJ0273. 【Conclusion】Rapeseed seedlings are very sensitive to herbicide ZJ0273 at germination stage. Treatment at 10 mg?L-1 ZJ0273 can significantly inhibit rapeseed growth and root viability, and the inhibitive effect is intensified with the increase of treatment concentrations and durations. The application of 1 mg?L-1 (critical concentration) is safe for rapeseed seedling growth.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Study on Conditions in Solid-State Fermentation of Rapeseed Meal by Mixed Strains for Amino Acid Fertilizer
    WANG Yong-hong,RAN Wei,ZHANG Fu-guo,YANG Xing-ming,XU Yang-chun,SHEN Qi-rong
    Scientia Agricultura Sinica. 2009, 42(10):  3530-3540 .  doi:10.3864/j.issn.0578-1752.2009.10.0019
    Abstract ( 1155 )   PDF (629KB) ( 1018 )   Save
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    【Objective】 An optimization of parameters in solid-state fermentation of rapeseed meal by mixture of two protease-producing bacterial strains Stenotrophomonas maltrophilia G12 ( FJ211222) and Bacillus pumilus K11 (FJ211221) for an amino acid fertilizer was studied to provide a theoretical basis in the industrial production. 【Method】 The fermentation processes were optimized by using of Response Surface Methodology (RSM) with Plackett-Burman Experiments and Box-Behnken Design. Data were analyzed with Design-Expert 7.0 software and Mintab 15.0 software. 【Result】 High amino acid production in fermentation processes was observed under the optimized condtions: 6 days of fermenting, 54.5% of initial moisture, initial pH value at 9.15, 14.1% of wheat bran supplementation, 5% of mixed inocula in equal volume, and one time of ventilation in 24 hours. 【Conclusion】 Under the optimized fermentation conditions, the degree of hydrolysis of rapeseed meal protein reached 13.1% with high contents of free amino acids and small peptides.

    Effects of Long-Term Application of Fertilizer and Rice Straw on Soil Fertility and Sustainability of a Reddish Paddy Soil Productivity
    LIAO Yu-lin,ZHENG Sheng-xian,NIE Jun,LU Yan-hong,XIE Jian,YANG Zeng-ping
    Scientia Agricultura Sinica. 2009, 42(10):  3541-3550 .  doi:10.3864/j.issn.0578-1752.2009.10.0020
    Abstract ( 1083 )   PDF (462KB) ( 995 )   Save
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    【Objective】 The effect of long-term application of nitrogen (N), phosphate (P) and potassium (K) fertilizer and rice straw (RS) on change trends of soil fertility and productivity sustainability of double rice (Oryza Sativa L.) cropping system were investigated in a reddish paddy soil of subtropical climate region in China. 【Method】 This study evaluated the soil fertility and productivity sustainability in five treatments composed of N, P and K fertilizer and combined application of fertilizer with RS in a long-term experimental field established in 1981 designed to examine the effects of fertilization on soil properties and rice crop productivity of a reddish paddy soil (classified as typical stagnic Anthrosol) derived from parent materials of Quaternary red clay. 【Result】 Results showed that long-term balanced application of N, P and K fertilizers and in combination with RS could increase significantly soil organic matter (SOM), total N (TN), total P (TP), potentially mineralizable N (PMN), available P and K compared with their initial value in 1981. Rice yield, N and K content in soil were lowered year by year in NP treatment without K fertilizer, and productivity sustainability of rice crop and soil fertility could not be maintained. Rice straw returned into soil significantly increased rice yield. Average rice yield with 27 years rice straw-returned treatment (NP+RS and NPK+RS) were increased by 12.2% and 6.7%, respectively, compared with NP and NPK application alone. 【Conclusion】 Rice straw could partly replace chemical K fertilizer as the nutrient of K brought by rice straw had the same function as that of chemical K fertilizer.

    Effects of Rice-Based Cropping System, Application Rate of Organic Manure and Ground Water Level on Inorganic Phosphorus Forms in Paddy Soil Derived from Red Earth
    FENG Yue-hua,ZHANG Yang-zhu,HUANG Yun-xiang
    Scientia Agricultura Sinica. 2009, 42(10):  3551-3558 .  doi:10.3864/j.issn.0578-1752.2009.10.0021
    Abstract ( 958 )   PDF (327KB) ( 620 )   Save
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    【Objective】 To ascertain the effects of rice-based cropping system, organic manure and ground water level on soil inorganic phosphorus forms. 【Method】 A long-term stationary experiment was carried out to study this subtect based on the fractional schemes proposed by Jiang Bai-fan, et al. 【Result】 Results showed that the contents of total inorganic phosphorus of the tested plough layer soils in different treatments ranged from 398.56 to 546.10 mg?kg-1, with (471.4±40.5) mg?kg-1 on average. O-P and Fe-P were the two main forms of inorganic phosphorus in the reddish-yellow clayey paddy soi1 that was derived from quaternary red clay and the two fractions accounted for 39.20%-38.59% of total inorganic phosphorus. Among the 3 rice-based cropping systems, the contents of total inorganic phosphorus in the treatment of rice-rice-flooded fallow in winter were higher than those in the treatments of rice-rice- rapeseed and rice-rice-green manure. The total inorganic phosphorus contents in the treatment of inorganic fertilizer were higher than those in the treatments of high rate of applied of manure and normal rate of applied manure. Low ground water level can increase the inorganic phosphorus contents in the soil. The contents of Fe-P in the treatments of rice-rice-flooded fallow in winter and rice-rice-rapeseed were higher than those in the treatments of rice-rice-green manure. As time goes on, the contents of Ca2-P, Ca10-P and Fe-P decreased under the same rice-based cropping system, while the contents of Al-P, Ca8-P and O-P increased. The contents of Ca10-P, Fe-P and Al-P in the treatment of inorganic fertilizer were higher than those in the treatments of normal rate of applied manure and high rate of applied manure. As time goes on, the contents of Ca2-P, Ca10-P and Fe-P decreased under the same organic manure application, while the contents of Al-P, O-P and Ca8-P increased. The contents of Ca10-P and Fe-P in the treatment of low ground water level were higher than those in the treatment of high ground water level, while the contents of Ca2-P and O-P in the treatment of high ground water level were lower than those in the treatments of low ground water level. As time goes on, the contents of Ca2-P and Fe-P decreased under the same ground water level, while those of Al-P, O-P and Ca8-P increased 【Conclusion】Crop planting in winter can decrease inorganic P content, while green manure application in winter can decrease Fe-P content. Inorganic fertilizer application could increase total inorganic P and Ca10-Pand Fe-P and Al-P contents. Lower ground water level is beneficial to accumulation of inorganic P.

    Application of Weighted Spatially Fuzzy Dynamic Clustering Algorithm in Evaluation of Soil Fertility
    CHEN Gui-fen,CAO Li-ying,WANG Guo-wei
    Scientia Agricultura Sinica. 2009, 42(10):  3559-3563 .  doi:10.3864/j.issn.0578-1752.2009.10.0022
    Abstract ( 1030 )   PDF (369KB) ( 812 )   Save
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    【Objective】 As a traditional fuzzy clustering algorithm has its shortages, an improved algorithm was presented in the paper. 【Method】 First, access to the weighted value of each attribute using AHP, and then add weighted value to the spatial fuzzy dynamic clustering algorithm, finally, use F- distribution of probability statistics to determine the best classification number in order to improve the algorithm intelligence. 【Result】 The weighted spaces fuzzy dynamic cluster algorithm was compared with the fuzzy equivalent relations transitive closure algorithm, the result shows that F-value is the largest and the classification results is best when λ=0.993. This time, the weighted F- value is 4.898, unweighted F- value is 2.957, that shows the weighted gap between class are more obvious that unweighted one, that is, the accuracy of the clustering algorithm is significantly higher than unweighted fuzzy clustering algorithm. 【Conclusion】 Tests show that the clustering algorithm’s accurate rate is higher than the unweighted fuzzy clustering algorithm.

    Influence of Tillage Patterns on Incorporated Straw Nitrogen Release and Nitrogen Utilization of Rice
    LIANG Tian-feng,XU Shi-hong,LIU Kai-qiang,WANG Dian-jun,LIANG He,DONG Deng-feng,WEI Shan-qing,MO Run-xiu,ZENG Ke,JIANG Li-geng
    Scientia Agricultura Sinica. 2009, 42(10):  3564-3570 .  doi:10.3864/j.issn.0578-1752.2009.10.0023
    Abstract ( 981 )   PDF (275KB) ( 1088 )   Save
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    【Objective】 The objective of this study was to investigate the difference of nitrogen release from incorporated rice straw and nitrogen utilization of rice under different tillage patterns. 【Method】 Rice straw was incorporated into field under no-tillage and conventional tillage in the late seasons of 2006 and 2007 and nitrogen contents in incorporated straw were measured at different growth stages after incorporation. At the same time, rice straw with 15N feed was incorporated into pot under no-tillage and conventional tillage, nitrogen and 15N uptake by rice plant were measured at maturity. 【Result】 Residual nitrogen amount in incorporated straw decreased and obvious differences were observed at different stages. The nitrogen loss in incorporated straw was about 60% during the 0-40 days and about 10% during the 40-100 days. However, the residual nitrogen content increased in two tillage patterns. Nitrogen in straw was more easily released under no-tillage than conventional tillage if the straw status was not considered. Nitrogen in wet incorporated straw was more easily released under no-tillage while dry straw under conventional tillage. 【Conclusion】 Great nitrogen dry matter production efficiency (NDMPE) and grain yield were observed under conventional tillage. No tillage was beneficial to straw nitrogen release, but nitrogen uptake from incorporated straw by rice plant and nitrogen use efficiency were decreased.

    HORTICULTURE

    Construction of Tomato Molecular Genetic Map and QTL Analysis of Resistance of Gene Cluster ph-3 to Tomato Phytophthora infestans

    HUANG Xiao-mei,XU Xiang-yang,LI Jing-fu,CHEN Xiu-ling,XU Yan-hui
    Scientia Agricultura Sinica. 2009, 42(10):  3571-3580 .  doi:10.3864/j.issn.0578-1752.2009.10.0024
    Abstract ( 1205 )   PDF (1212KB) ( 905 )   Save
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    【Objective】 Late blight is a worldwild disease in tomato and can cause serious yield loss. Through construction of tomato molecular genetic map and QTL analysis of gene cluster ph-3 to find closely linked molecular markers. This will provide a theoretical and practical basis for molecular marker-assisted breeding. 【Method】 L3708(L. pimpinellifolium)accompanied with resistant gene cluster ph-1,2,3 was used as male parent and 04968(L. esculentum)susceptible race with good characters as female parent, the 260 F2 single plants as construction population, a molecular marker linkage map composed with 12 linkage populations was constructed by SSR and AFLP molecular marker. QTLs were found by applying complex-space mapping and according to seedling-inoculated tomato’s resistant response to Phytophthora infestans physiological stains T1, 2. 【Result】 The map includes 3 SSR markers and 149 AFLP makers. And the map coveres 1 443.07 cM of the whole genome. The average space between markers was 9.50 cM. Five QTLs correlative to ph-3 were found. Qph3-1 was on the first chromosome, explainable phenotype variation was 26.59%. Qph3-2 was on the sixth linkage population and the explainable phenotype variation was 54.86%. Qph3-3, Qph3-4, Qph3-5 were on the ninth linkage population and the explainable phenotype variations were 9.24%, 10.27%, and 36.49%. The genetic effect of QTL loci was additive and dominant. 【Conclusion】 The 5 QTL screened in this study can be used as a tool for molecular marker-assisted breeding.

    Construction the Engineered Restoring Line of Tomato Engineered Male Sterile Line by Cre/lox Site-Specific Recombination System
    SONG Hong-Yuan,REN Xue-Song,SI Jun,LI Cheng-qiong,SONG Ming,LEI Jian-Jun
    Scientia Agricultura Sinica. 2009, 42(10):  3581-3591 .  doi:10.3864/j.issn.0578-1752.2009.10.0025
    Abstract ( 936 )   PDF (961KB) ( 925 )   Save
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    【Objective】 An engineered restoring line of tomato engineered male sterile line was constructed using Cre/lox site-specific recombination system, which restore the fertility by deleting the male sterile gene directly in F1. 【Method】 The expression vector of pBinBarloxTABn, contained a TA29-Barnase cassette flanked by two lox sites in a directed orientation, a NPTⅡ gene expression cassette and a Bar gene expression cassette was introduced into the tomato genome to bring male sterile line. The Cre gene, under the control of CaMV 35S promoter, was transformed into tomato to construct the engineered restoring line. The Cre gene was introduced into the F1 generation by pollinating the tomato male sterile plants with pollen from Cre-expression plants, expression of the Cre in the hybrid leads to the removal of the TA29-Barnase gene, then the plants was fertile. 【Result】 The tomato male sterile plants were obtained by Agrobacterium tumefaciens-mediated transformation, using the NPTⅡ gene as the transformation maker gene. The Bar gene in the male sterile plants expressed well, and these plants showed considerable resistance to herbicide Basta, the leaf disc differentiated callus and shoots on the medium supplemented with PPT 3 mg?L-1, and kept fresh in the solution containing PPT 20 mg?L-1 in 6-7 days. The male sterile plants characterized stamen degenerate, no pollen or tiny innormal pollens- without vigor. No normally expanded fruits and formed seeds were observed in the transgenic plant after self-pollination of the male sterile plants. However, the normally expanded fruits and seeds were observed after cross-pollination of the male sterile plants using pollens from wild-type plant, and the progenies from the hybrid showed a 1﹕1 ratio of Basta resistant to Basta susceptive. Cross-pollinated the male sterile plants (T0) using pollens from Cre-expression plants, the normally expanded fruits and seeds were observed. The molecular analysis on the progenies from the hybrid was performed, the results showed that those progenies inherited both the Cre gene and Bar gene had lost the TA29-Barnase gene without exception, and those progenies could flower and fruit normally, indicated that the male sterility had been restored. 【Conclusion】 Using the Cre/lox site-specific recombination system, the male sterility-causing gene integrated into the tomato genome was successfully eliminated by crossing with pollen from a engineered restoring line expressing the Cre recombinase, yielding hybrid fruits and seeds. This results have provided an alternative method for the fertility restoration of plant engineered male sterility.

    Effects of Night Low Temperature on Sugar Accumulation and Sugar-Metabolizing Enzyme Activities in Melon Fruit
    HAO Jing-hong,LI Tian-lai,MENG Si-da,ZHAO Bo,SUN Li-ping
    Scientia Agricultura Sinica. 2009, 42(10):  3592-3599 .  doi:10.3864/j.issn.0578-1752.2009.10.0026
    Abstract ( 921 )   PDF (420KB) ( 1238 )   Save
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    【Objective】 The effects of different nigh low temperatures on carbohydrate metabolism in melon fruit were studied in this paper. 【Method】 Each carbohydrate content and sugar-metabolizing enzymes activities in fruits grew under normal and night low temperature circumstance simulated by climate room were analyzed. 【Result】 Effects of different night low temperatures on carbohydrate contents in melon fruit were different to some degrees. Fructose and glucose decreased after treatment of 9℃ in night for 3 d while significantly decreased sucrose content occurred in middle and later period, yet galactinol content had no change. Meanwhile, total carbohydrate content was lower than the control with markedly significant difference after treatment of 9℃ in night for 3 d, and the difference was more obvious as the time prolonging. Otherwise, compared with the control, starch content increased obviously on 3-12 d after treatment of 9℃ in night, however, after treatment of 12℃ in night, starch content increased in evidence while each carbohydrate content was almost invariable. Meanwhile, the change of sugar-metabolizing enzymes activities in fruit indicated that acid invertase (AI), netutral invertase (NI) and ?-galactosidase activities enhanced, but sucrose synthase (SS) activities and sucrose phosphate synthase (SPS) activities declined with very significant difference after treatment of 9℃ in night. What was more, the effects of 12℃ in night on sugar-metabolizing enzymes activities were small, with no significant difference. 【Conclusion】 Sugar accumulation and sugar-metabolizing enzymes activities were affected seriously after treatment of 9℃ in night, but 12℃ in night did not altered the carbohydrate metabolism, which was the critical low temperature to restrict sugar accumulation. These results suggested that the decreased metabolism including sucrose, stachyose and starch may affect melon fruit expansion and quality after treatment of 9℃ in night.

    Cloning and Expression of Calcium-Dependent Protein Kinase (CDPK) Gene Family in Nicotiana tabacum
    TAI Shuai-shuai,LIU Guan-shan,SUN Yu-he,CHEN Jia
    Scientia Agricultura Sinica. 2009, 42(10):  3600-3608 .  doi:10.3864/j.issn.0578-1752.2009.10.0027
    Abstract ( 1273 )   PDF (705KB) ( 1372 )   Save
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    【Objective】To further study the function of calcium-dependent protein kinase (CDPK) gene family in tobacco (Nicotiana tabacum), it is necessary to isolate CDPKs from tobacco and describe the sequence characteristics, evolutionary relationship and gene expression. 【Method】 RT-PCR, RACE and bioinformatics methods were used to isolate CDPKs from tobacco. A phylogenetic tree was created using the MEGA4.0 program and the expression patterns of the 3 full-length CDPK genes were studied by RT-PCR. 【Result】 Eight additional tobacco CDPK genes were obtained, of which 3 possessed complete ORFs. Phylogenetic analysis divided 11 full-length Nicotiana CDPK genes into 4 subfamilies, and 2 putative tobacco and Arabidopsis orthologous CDPK genes might correspond to well-conserved functions. Three full-length tobacco CDPK genes were detected in all tobacco organs tested, but the expression patterns of them were significantly different. 【Conclusion】 Eight non-redundant tobacco CDPK genes were isolated in this study. Along with the previously characterized CDPK genes, at least 15 members of the CDPK family are existing in tobacco. This work established a foundation for a genome-wide study of this important gene family in tobacco.

    STORAGE·FRESH-KEEPING·PROCESSING
    Study on the Properties of Apple Pectin Isolated by Ultrafiltration
    QIU Nong-xue,TIAN Yu-xia,DENG Hong,GUO Yu-rong,QIAO Shu-tao
    Scientia Agricultura Sinica. 2009, 42(10):  3609-3616 .  doi:10.3864/j.issn.0578-1752.2009.10.0028
    Abstract ( 1011 )   PDF (426KB) ( 992 )   Save
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    【Objective】 Separation conditions of ultrafiltration for apple pectin were investigated. The relationship between different molecular weights of apple pectin and their structures and properties were also determined. 【Method】 Five ultrafiltration membranes in different molecular weight cut-off were applied to isolate apple pectin. FT-IR and GC were developed to determine the structures and monosaccharide composition of apple pectin with different molecular weights. 【Result】 Under ultrafiltration conditions of temperature 50℃, pressure 0.08 MPa and concentration 1 g?L-1, six kinds of pectins with different molecular weights were obtained. Meanwhile, molecular weight of pectin was related to its physicochemical properties. The larger the molecular weight of pectin was, the more its galacturonic acid content and degree of esterification and gelatination were. In addition, ultrafiltration contributed to the decolouring of pectin extract. 【Conclusion】 Ultrafiltration could isolate apple pectin well and it could be a new method of purification, separation and development for apple pectin.

    Effect of Exogenous Acetaldehyde on Reactive Oxygen Metabolism in Medium-Term and Late Maturity Peaches During Storage
    LI Jian-li,HAN Tao,LI Li-ping,SUN Yu-jing,ZHANG Wei-na,ZHANG Hai-ying
    Scientia Agricultura Sinica. 2009, 42(10):  3617-3624 .  doi:10.3864/j.issn.0578-1752.2009.10.0029
    Abstract ( 925 )   PDF (382KB) ( 611 )   Save
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    【Objective】 The effects of exogenous acetaldehyde treatment on reactive oxygen metabolism in peaches (Prunus persica L.)during storage were investigated. 【Method】 The medium-term maturity of ‘Okuba’ and late maturity of ‘Lühua 9’ peaches were fumigated with 0, 0.25, 0.50 and 1.00 ml?L-1 acetaldehyde (AA) for 12 hours,then stored at room (20±1℃) or low (0±1℃) temperatures. The active oxygen metabolism of the fruits during storage was investigated. 【Result】At the two temperatures 1.00 ml?L-1 AA treatment increased the activities of superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD) of peach in various extents, decreased the production rate of superoxide anion radical ( ) and the contents of hydrogen peroxide (H2O2) at different periods of storage. AA treatment at 0.50 ml?L-1 also decreased the production rate of and the contents of H2O2, increased the activities of SOD and POD but the light influence on activities of CAT at 20℃. 0.50 ml?L-1 treatment showed a light influence on the active oxygen metabolism. AA treatment at 0.25 ml?L-1 had little influence on the active oxygen metabolism. There were no obvious differences in the parameters above among the treatments of fruits during the shelf life. 【Conclusion】 Reactive oxygen metabolism in peaches was held back by exogenous acetaldehyde treatments at suitable concentration at room temperature (20±1)℃and (0±1)℃, respectively.

    Effect of Pre-Slaughter Conditions on Beef Quality
    DU Yan,ZHANG Jia,HU Tie-jun,LUO Xin
    Scientia Agricultura Sinica. 2009, 42(10):  3625-3632 .  doi:10.3864/j.issn.0578-1752.2009.10.0030
    Abstract ( 988 )   PDF (373KB) ( 1165 )   Save
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    【Objective】 The objective of this paper was to study the beef quality which had different pre-slaughter managements, to educe the judgment criterion of DFD (dark firm dry) beef. 【Method】 The incidence of DFD beef in one corporation was studied, then 600 bulls were assessed by measuring pH and temperature values in the Longissimus dorsi muscle and studying the beef quality, the incidence and the quality of inferior beef were analyzed. 【Result】 Results revealed that pre-slaughter condition which included the stocking density, temperature and humidity of lairage and the short distance transport had a little effect on the incidence of DFD beef. However, slaughter time, season, lairage time, and slaughter method had an effect on the incidence of inferior beef (PSE and DFD meat), more or less; b* which is the variable of beef color was more important than L* to estimate inferior beef. To estimate the DFD beef, the criterion is pH24 value>6.09, L*<37.40±3.78, b*<10.25±1.94, drip losses<7.36%±2.81%. 【Conclusion】 The incidence of DFD meat was lower when the bull had experienced 24 h lairage and stunned slaughter method in winter morning. In summer, the incidence of PSE beef was higher when the bull had experienced Islam slaughter method. The stress reaction of animal and the incidence of inferior beef decreased obviously by scientific and normative pre-slaughter managements. When pH24 value>6.09, the beef quality changes greatly.

    ANIMAL SCIENCE·RESOURCE INSECT
    Effect of LH on Nuclear and Cytoplasm Synchronization for in vitro Maturation of Ovine Oocytes
    LI Kai,NI He-min,ZHAO Yan-wei,LIU Yun-hai,QU Yang-yan,LI Xiao-yan,GUO Yong
    Scientia Agricultura Sinica. 2009, 42(10):  3633-3638 .  doi:10.3864/j.issn.0578-1752.2009.10.0031
    Abstract ( 995 )   PDF (364KB) ( 790 )   Save
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    【Objective】 This study was made to investigate the effect of LH on maturation and fertilization in vitro of ovine oocytes. 【Method】 The ovine oocytes were matured in vitro with or without LH, and checked by confocal laser scanning microscope to observe the distribution of cortical granules during different durations stained with FITC-LCA. Meanwhile, some in vitro matured oocytes were also fertilized in vitro for analysis of their developmental potentiality further. 【Result】 After 4 h of culture, there were significant differences in the rate of GVBD between the treatment (with LH) and control (without LH) groups (36.76% vs 50%, P<0.05), moreover, there were also significant differences in cleavage rate and blastocyst rate, respectively, between these two groups (67.15% vs 42.37%, 21.9% vs 12.71%, P<0.05). The distribution of cortical granules appeared to spread from the edges to the central site of sheep oocytes following their in vitro maturation durations. 【Conclusion】 LH plaies a role to delay the occurrence of germinal vesicle, prolong the maturation duration of cytoplasm,enhance the nuclear and cytoplasm synchronization of ovine oocytes matured in vitro, and finally improve the potentiality of their in vitro development.

    Correlation Analysis Between Microsatellite Markers and Milk Performance Traits in Laoshan Dairy Goat
    LI Cheng-bo,WANG Gui-zhi,JI Zhi-bin,GE Xin,LI Pei-pei,WANG Jian-hua,WANG Jian-min
    Scientia Agricultura Sinica. 2009, 42(10):  3639-3646 .  doi:10.3864/j.issn.0578-1752.2009.10.0032
    Abstract ( 1002 )   PDF (766KB) ( 893 )   Save
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    【Objective】 The aim of the present work was to research whether the microsatellite DNA could improve milk performance traits of Laoshan dairy goat as the genetic marker. 【Method】 According to comparative genetic map, five dairy cattle microsatellite loci (CSSM66, BM302, BM1329, BM143 and BMS0812) correlated with milk performance traits of chromosome 4, 6 and 15 were detected in 175 Laoshan dairy goats. The genetic analysis and correlation analysis between microsatellite markers and milk performance traits were calculated in Laoshan dairy goat. 【Result】 The genetic analysis showed that 5 microsatellite loci had a rich diversity in Laoshan dairy goat group. The mean values of the Ne, Ho, PIC and Fis were 0.39924, 0.4667, 0.6748 and 0.347 respectively. Least square analysis evidenced that CSSM66 had a significant effect on the fat percentage (P<0.01) and 300 d milk yield (P<0.05), and the genotypes 210/210 and 194/194 were the most favorable genotypes respectively, the alleles 210 and 194 had a positive effect on fat percentage and 300 d milk yield respectively. BM302 had a significant effect on 300 d milk yield(P<0.01), and the genotype 156/150 was the most favorable genotype, the alleles 156 and 150 had a positive effect on 300 d milk yield. BM143 had a significant effect on the fat percentage (P<0.05) and protein percentage (P<0.01), and the genotypes 114/100 and 100/100 were the most favorable genotypes, both the alleles 114 and 100 had a positive effect on fat percentage and protein percentage. BM1329 had a significant effect on fat percentage (P<0.05), and the genotype 182/180 was the most favorable genotype. 【Conclusion】 In this study, the microsatellite loci with a rich diversity and a significant effect on milk performance traits were found, and they provided valuable genetic markers for marker-assisted selection of Laoshan dairy goat.

    Correlation Between Peroxisome Proliferator-Activated Receptor γ 5'Flanking- Region Haplotypes with the Growth and Body Composition Traits in Chickens
    HAN Qing,WANG Shou-Zhi,HU Guo,LI Hui
    Scientia Agricultura Sinica. 2009, 42(10):  3647-3654 .  doi:10.3864/j.issn.0578-1752.2009.10.0033
    Abstract ( 1156 )   PDF (413KB) ( 779 )   Save
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    【Objective】 The chicken PPARγ gene 5' flanking region was used to detect the polymorphisms, and the correlation between the polymorphism and growth and body composition traits were analyzed in order to find the quantitative trait locus which providing a basis for molecular marker-assisted selection. 【Method】The 8th to 10th generation populations of the Northeast Agricultural University broiler lines divergently selected for abdominal fat content were used. Growth and body composition traits were measured in the above populations. Polymorphisms among individuals were detected by DNA sequencing, PCR-RFLP and some other methods. The correlation analysis between the haplotypes constructed with the three polymorphisms detected and the growth and body composition traits was performed. 【Result】 The correlation analysis showed that haplotypes, based on three polymorphisms in 5' flanking region of PPARγ gene (g.-1784_-1768del17, c.-1241G>A and c.-75G>A), were significantly correlated with abdominal fat weight (AFW), abdominal fat percentage (AFP), liver weight (LW), liver weight percentage (LW/BW), shank length (ShL), femur weight (FeW), feel length (KeL), metatarsus circle (MeC) (P<0.05), and suggestively significantly correlated to some extents to pectoralis major weight (PMaW), pectoralis minor weight (PMiW), pectoralis minor weight percentage (PMiW/BW), metatarsus length (MeL) (P<0.2). The least square analysis showed that the birds with BGA haplotype had significantly higher AFW and AFP than the birds with other haplotypes (P<0.05). The birds with AAG haplotype had significant higher LW and LW/BW than the birds with other haplotypes (P<0.05). The birds with AAG haplotype had significant higher PMiW and PMiW/BW than the birds with other haplotypes (P<0.05). The birds with AAG haplotype had significant higher ShL, FeW, MeL, MeC and KeL than the birds with AGG haplotypes (P<0.05). 【Conclusion】 The haplotype-based correlations reveal that there may exist QTL in 5' flanking region of PPARγ, affecting abdominal fat traits in chickens, and PPARγ gene might be one of the genes having an important influence on the growth and skeletogenesis traits in chickens.

    Relationship Between the BMP2, BMP4, BMP6 and BMP7 Gene Expression and Ovulation Number in Hu Sheep
    XU Ye-fen,LI Qi-fa,LI Er-lin,TU Fei,HU Dong-li,XIE Zhuang,CHEN Ling
    Scientia Agricultura Sinica. 2009, 42(10):  3655-3661 .  doi:10.3864/j.issn.0578-1752.2009.10.0034
    Abstract ( 1077 )   PDF (487KB) ( 791 )   Save
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    【Objective】 The present study was performed to investigate the relationships between the mRNA level of BMP2, BMP4, BMP6 and BMP7 gene in ovaries and the ovulation number in Hu sheep, so as to provide references for finding hyper- prolificacy candidate gene and understanding the molecular genetic mechanism of high fecundity in Hu sheep. 【Method】 Sixteen adult Hu ewes, classified as the single lambling (S) or multiple lambling (M) group, were sacrificed for tissue sampling and ovulation number counting between 24-36h after estrus diagnosis. The tissue distributions of BMP2, BMP4, BMP6 and BMP7 mRNA were detected by RT-PCR and the gene expression levels in the ovaries were investigated by real-time PCR. 【Result】 BMP2, BMP4 and BMP7 mRNA exited in the ovary and other tissues, including hypothalamus, pituitary, uterus, heart, liver, spleen, lung, kidney, muscle and oviduct, but BMP6 mRNA was only detected in the ovary, kidney, muscle and oviduct. The expression level of BMP2, BMP6 and BMP7 mRNA in the ovary did not differ between S and M group (P>0.05), but BMP4 mRNA abundance in the ovary was extremely higher in M group than that in S group (P<0.01). Bivariate correlation analysis showed that BMP4 mRNA expression in the ovary was positively correlated with ovulation number (r = 0.741, P<0.05), but there was no correlation between the mRNA expression level of BMP2, BMP6 or BMP7 and the ovulation number (P>0.05). 【Conclusion】 The results suggested that BMP4 might play a key role in ovulation number and might be a candidate gene for high fecundity in Hu sheep.

    Identification of Swainsonine-Producing Fungal Endophytes from Three Species of Locoweeds in Tibet
    YU Yong-tao,WANG Jian-hua,WANG Yan,SONG Yu-min,GENG Guo-xia,LI Qin-fan
    Scientia Agricultura Sinica. 2009, 42(10):  3662-3671 .  doi:10.3864/j.issn.0578-1752.2009.10.0035
    Abstract ( 1047 )   PDF (528KB) ( 765 )   Save
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    【Objective】 The fungal endophytes were isolated from three locoweeds Oxytropis sericopetala, Oxytropis glacialis and Astragalus strictus in Tibet. 【Method】 The swainsonine isolated from the endophytes was analyzed with thin-layer chromatography, gas chromatography and gas chromatography-mass spectrometry. The ITS and IGS regions of fungi were amplified by polymerase chain reactions. In addition to the sequences of ITS regions from endophytes, the phylogenetic tree based on the ITS sequences was constructed to determine the taxon of the endophytic fungi. The amplified products of IGS regions from endophytes were digested using restriction enzymes and the digested products were analyzed. 【Result】 Four swainsonine-producing fungal endophytes were isolated from three species of locoweeds. The morphology of fungi and homology of ITS sequences was closely related to that of Embellisia spp. Four isolates were named Embellisia oxytropis based on the phylogenetic tree. 【Conclusion】 Analysis of the ITS sequences and the digested IGS fragments indicated that four endophytes were closely related species. However, the intraspecific variation was present among the fungal endophytes from different host plants and the isolates may be different subspecies of Embellisia oxytropis. The research is important to explore the SW biosynthesis mechanism of endophytes and has provided a novel knowledge for the elimination of animal locoism and a reasonable utilization of locoweeds.

    VETERINARY SCIENCE
    Construction of Recombinant Lactococcus Lactis Expressing VP4 of Porcine Rotavirus and Analysis of Immunogenicity
    LI Gui-wei,QIAO Xin-yuan,LIU Bo-chen,MA Guang-peng,LIU Min,LIU Li-wei,LI Yi-jing
    Scientia Agricultura Sinica. 2009, 42(10):  3672-3678 .  doi:10.3864/j.issn.0578-1752.2009.10.0036
    Abstract ( 1016 )   PDF (462KB) ( 712 )   Save
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    【Objective】 The main protective antigen gene segment VP4 of porcine rotavirus was inserted into the surface display vector pNZ8112 of Lactococcus. 【Method】 It was electrotransformed into Lactococcus lactis NZ9000, and the surface display system pNZ8112/NZ9000 was construced. To prove the expression of the exogenous gene, the expression product was detected by SDS-PAGE, Western-blot and immunofluorescence.The reconbinant strain was orally administered to the 6-8 week Balb/c mice. 【Result】 The regional humoral immunity and systematic immunity were detected in the immunized mice.The results indicate that specific sIgA and IgG can be found in fecal pellet, ophthalmic wash, vaginal wash and in the serum after administration.The result of neutralization test indicate the titer of serum is 1:40. 【Conclusion】 All the results indicate that VP4 expressed in the recombinant strain can elicit regional immunity and systematic immunity.

    Effects of AvBD13 on Humoral Immunity of Chickens
    YANG Yu-rong,SHE Rui-ping,LIANG Hong-de
    Scientia Agricultura Sinica. 2009, 42(10):  3679-3684 .  doi:10.3864/j.issn.0578-1752.2009.10.0037
    Abstract ( 929 )   PDF (418KB) ( 638 )   Save
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    【Objective】 The aim of this study was to investigate the effect of AvBD13 on the humoral immune response of chicken. 【Method】 In the treatment group chickens received drinking water supplemented with AvBD13 (1 μg?ml-1) right after hatching. Samples of blood, bursa of Fabricus, spleen and intestine were taken at day 1, 4, 7, 10 and 17 of experiment. B lymphocyte proliferation in peripheral blood stimulated by mitogen,the content of immunoglobulin in serum, antibody-forming cells in immune organ and intestine were measured by MTT, ELISA or immunohistochemistry methods. 【Result】 AvBD13 supplementation enhanced the content of IgG and IgM in serum from day 4-10 and day 10-17, respectively (P<0.05), IgM-forming cells in bursa of Fabricus and spleen at the age of 4-7 days (P<0.05). In addition, AvBD13 could slightly enhance the number of IgM-forming cells and IgG-forming cells in intestine at day 4 to 10, but had no significant difference (P>0.05), AvBD13 only enhanced the cecal tonsils diffuse area IgA-forming cells at day 4 (P<0.05). 【Conclusion】 These results suggested that AvBD13 could greatly enhance systemic humoral immune responses and slightly enhance mucosal humoral immune responses, which maybe the reason of tissue specific effect of defensins.

    Cloning, Expression and Bioactivity Characterization of Duck Avian β-Defensin 2
    WANG Rui-qin,LIAO Wen-yan,MA De-ying,HAN Zong-xi,LIU Sheng-wang
    Scientia Agricultura Sinica. 2009, 42(10):  3685-3692 .  doi:10.3864/j.issn.0578-1752.2009.10.0038
    Abstract ( 991 )   PDF (436KB) ( 695 )   Save
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    【Objective】 The objective of the study was to clone avian β-defensin (AvBD) 2 gene from duck tissues, expression the recombinant AvBD2 protein in E.coli, and determine its antimicrobial activity. 【Method】 The mRNA of duck AvBD2 was cloned from pancreas of duck by RT-PCR. In addition, phylogenetic relationships between amino acid sequence of the duck AvBD2, AvBDs from other avian species, and some mammalian beta-defensin-2 were analyzed. The cDNA of duck AvBD2 was sub-cloned into pGEX-6p-1 vector to construct recombinant plasmid pGEX-duck AvBD2. The recombinant protein was expressed into E. coli. and purified. Antimicrobial activity and physical-chemical stability of the recombinant fusion protein was measured in vitro. 【Result】 The sequence analysis showed that the full length cDNA of duck AvBD2 consisted of 195 bp, encoding 64 amino acid residues. Homology analysis showed that duck AvBD2 shared high amino acid homology with AvBD2 from other avian species. A high expression level of the recombinant duck AvBD2 was noted in E. coli (molecular weight, 32 kD). The recombinant protein exhibited high antimicrobial activity against Bacillus subtilis, Staphylococcus aureus, and Pasteurella multocida, low antimicrobial activity against E. coli., and Salmonella. choleraesuis. The minimal inhibitory concentration (MIC) of the recombinant protein against the above bacteria ranged from 15.25-125 μg?ml-1. The MIC of the recombinant protein against S.choleraesuis was>400 μg?ml-1. In addition, the results revealed that the recombinant protein retained antimicrobial activity against S. aureus under different temperatures (-20℃ to 100℃) and pH values (3 to 12). 【Conclusion】The AvBD2 gene from ducks was successfully cloned, expressed in E. coli, and purified. The recombinant protein showed antimicrobial activity and retained stability under different temperatures and pH values.

    Transfection Technology and Its Application in the Investigation of Apicomplexan Protozoa
    YAN Wen-chao,WANG Tian-qi,SUO Xun,ZHAO Zhan-qin,HAN Li-fang
    Scientia Agricultura Sinica. 2009, 42(10):  3693-3699 .  doi:10.3864/j.issn.0578-1752.2009.10.0039
    Abstract ( 930 )   PDF (295KB) ( 1010 )   Save
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    Since transient transfection of Toxoplasma gondii was first reported in 1993, stable transfections of apicomplexa such as T. gondii, Plasmodium species and Sarcocystis neurona have been achieved later, moreover, both the in vitro transient and in vivo stable transfection systems of Eimeria tenella, which do not undergo re-cycling of any life cycle stages, has been developed successfully. With the rapid development of transfection technology in Apicomplexan parasites, genetic manipulation provides a powerful tool for the study of the biochemical and cell biology of Apicomplexa. This paper has reviewed the latest progress of transfection technology and its application in gene regulation, the function of gene, the nature of drug resistance, drug sensitivity test and immunology of Apicomplexan parasites in combination with the related researches in authors’ laboratory in past years.

    Effects of Conglycinin Peptides on Development of Bacterial Community in Faeces of Rats
    ZUO Wei-yong,CHEN Wei-hua,ZOU Si-xiang,ZHANG Yuan-shu
    Scientia Agricultura Sinica. 2009, 42(10):  3700-3705 .  doi:10.3864/j.issn.0578-1752.2009.10.0040
    Abstract ( 1012 )   PDF (423KB) ( 621 )   Save
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    【Objective】 The objective of this experiment is to investigate the effect of conglycinin peptides on bifidobacteria in vivo under the molecular level.【Method】 Twelve male SD rats were randomly assigned into 4 groups, and each group was administered for 21d intragastrically with physiological saline (control group), conglycinin (conglycinin group), the most active fraction which isolated from pepsin-treated conglycinin (P2-PTC group) and HCl-full hydrolysis of conglycinin (HCl-FHC group) respectively, 0.5 ml volume of administration with equal amount of nitrogen (0.5 mg?ml-1) for each rat. PCR and denaturing gradient gel electrophoresis (DGGE) were used to monitor the development of bacterial community in faeces of rats which were administered intragastrically with conglycinin peptides. The regions V6 to V8 of the 16S rDNA of faecal bacteria were amplified. DGGE profiles of the PCR amplicons were compared by similarity analysis. 【Result】 Rats administered with conglycinin peptides, DGGE similarity analysis showed that the bacterial community in the faeces developed fast at the first week of experiment and relatively slow later. It suggested that conglycinin peptides were capable of modifying the bacterial composition of the caecal content in rats. To investigate the effect of conglycinin peptides on bifidobacteria in vivo, 16S rDNA gene sequence specific for Bifidobacterium in fecal digesta were quantitatively detected by series dillution PCR. It was found that the number of bifidobacteria was increased in conglycinin peptides group compared with the control and HCl-FHC groups. 【Conclusion】 The results showed that conglycinin peptides can increase the growth of bifidobacteria in vivo.

    RESEARCH NOTES
    Differentially Expressed Genes Between Cytoplasmic Male Sterility Lines and Maintainer Lines of Gossypium harknessii
    MA Yong,WU Jian-yong,XING Chao-zhu,GUO Li-ping,GONG Yang-cang,CUI Ming-hui,Wang Hai-lin
    Scientia Agricultura Sinica. 2009, 42(10):  3706-3712 .  doi:10.3864/j.issn.0578-1752.2009.10.0041
    Abstract ( 958 )   PDF (475KB) ( 744 )   Save
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    【Objective】 By identifying the differentially expressed transcripts of anthers at the critical period of abortion between cytoplasmic male sterility and maintainer lines of Gossypium harknessii, this study is to explore the possible molecular mechanism of the cytoplasmic male sterility, and lay a basis on utilizing cytoplasmic male sterility lines. 【Method】 cDNA amplified fragment length polymorphism analysis was used to isolate the differentially expressed fragments, these fragments were verified by reverse Northern blot, and then were cloned, sequenced and analyzed by bioinformatic tools. 【Result】 Totally, 108 differentially expressed fragments were identified, of which, 67 were succeeded in cloning. Homology searches performed by BLAST at web servers of NCBI showed that there were 29 fragments could be found homology sequences in GenBank database. By analyzing the Gene Ontology of these 29 sequences, it could be concluded that these sequences mainly participated in signal transducer, transcription, energy metabolism, cell wall development. Among them, some differentially expressed genes related to RNA edit and male gametogenesis were identified between two materials, and these maybe the mainl factors of male sterility. 【Conclusion】 The results may help to understand the relevant gene regulatory networks of cytoplasmic male sterility of cotton, and provide a pilot platform for further investigating the mechanism of CMS.

    Physiological Effects of Seed Priming on Salt Resistance of Sorghum Seedlings
    MA Jin-hu,WANG Hong-fu,WANG Yu-guo,LI Xin-ji,WEI Xian-guo
    Scientia Agricultura Sinica. 2009, 42(10):  3713-3719 .  doi:10.3864/j.issn.0578-1752.2009.10.0042
    Abstract ( 1199 )   PDF (217KB) ( 740 )   Save
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    【Objective】 Physiological effects of seed priming on salt resistance of sorghum seedlings were studied in this paper. 【Method】 Four sorghum hybrids seeds (Tiansi No. 1, Tx623A×89-363, Hei 30A×05-244 and Jinza 17) were soaked in 100 mmol?L-1 NaCl and 20%PEG solutions. Different degrees of salt injury were simulated with 0, 50, 100 and 150 mmol?L-1 NaCl solution by methods of nutritional sand culture. After seeds priming, germination conditions, salt resistance and physiological effects on seedlings were studied. 【Result】 Priming treatment enhanced the tolerance index and tolerance ratio to some degree. At the same time, the germination potential, germination rate, germination index and activity index were improved obviously, especially for the NaCl priming. In the course of stress of 100 mmol?L-1 NaCl solution, in particular the late stress, NaCl priming reduced MDA contents, enhanced SOD activities, contents of proline and soluble proteins in sorghum seedlings. 【Conclusion】 Seed priming technology could enhance the salt tolerance of sorghum seedlings which solved the problem of low germination rate and saving seedling rate in saline soil. The salt tolerance of sorghum seedlings was improved by increasing the activities of protective enzymes and contents of osmotic adjustment substances under the condition of salt stress.

    Inhibitory Effects of Two Compounds Extracted from Daphniphyllum calycinum on Phenoloxidase of Spodotera exigua
    LIU Wei,XIAO Ting,DU Lei,XUE Chao-bin,YANG Fan,LUO Wan-chun
    Scientia Agricultura Sinica. 2009, 42(10):  3720-3725 .  doi:10.3864/j.issn.0578-1752.2009.10.0043
    Abstract ( 1075 )   PDF (243KB) ( 827 )   Save
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    【Objective】 The article studies phenoloxidase inhibitors against insect and finds the clues on new target for pest control. 【Method】 The effects of deoxycalyciphylline B and Methyl homosecodaphniphyllate extracted from Daphniphyllum calycinum. on phenoloxidase (PO) of Spodoptera exigua (Hübner) were investigated adopting the microtitrationassay. 【Result】 The results indicated that the two compounds showed different inhibitory effects on the PO and the inhibitor concentrations leading to 50% (IC50) activity lost were estimated to be 2.439 mmol?L-1 for deoxycalyciphylline B and 0.879 mmol?L-1 for methyl homosecodaphniphyllate, respectively. The two compounds were also found to be reversible competitive inhibitors and the Ki values were determined to be 2.051 mmol?L-1 and 1.269 mmol?L-1, respectively. 【Conclusion】 Two alkaloids have better inhibitory activities on the PO. The two compounds or their analogues can be used as the templates to guide the molecular design of enzyme inhibitors. These are important ways looking for new pest control agents.

    Test and Preliminary Application of the SIMETAW Model in Northwest of Liaoning
    SHI Jin-li,WANG Li-gang,QIU Jian-jun,REN Tian-zhi,GENG Xu,YANG Xiao-guang,SUN Zhan-xiang
    Scientia Agricultura Sinica. 2009, 42(10):  3726-3733 .  doi:10.3864/j.issn.0578-1752.2009.10.0044
    Abstract ( 1143 )   PDF (340KB) ( 848 )   Save
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    【Objective】 The aim of this paper is to test the validity of SIMETAW (simulation of evapotranspiration of applied water) model developed by the University of California in Northwest of Liaoning Province, China. 【Method】 The comparison with model simulated and field measured evapotranspiration of maize, soybean and millet in northwest of Liaoning province were conducted, as well the analysis of the historical evapotranspiration and water demand of local main crops by applying the tested SIMETAW model were completed. 【Result】 The results showed that the model have good performances in simulating dynamic of main crops’ evapotranspiration intensity in northwest of Liaoning, and the simulated total evapotranspiration was very close to the field measured data, the relative error between the comparison was less than 10%. Based on the model simulation, the simulated multi-years’ average evapotranspiration for maize, soybean and millet during the growth stage were 514.15, 449.64 and 389.12 mm respectively, and the water demand for these three crop were respectively 208.4, 220.93 and 116.17 mm. 【Conclusion】 SIMETAW model has well validity on simulating the evapotranspiration of maize, soybean and millet in northwest of Liaoning province. The model can be further applied to water management study and guide irrigation in this region.

    Effect of Interaction Between Rootstock and Scion on Chilling Tolerance of Grafted Eggplant Seedlings Under Low Temperature and Light Conditions
    ZHANG Xiao-yan,XU Kun
    Scientia Agricultura Sinica. 2009, 42(10):  3734-3740 .  doi:10.3864/j.issn.0578-1752.2009.10.0045
    Abstract ( 961 )   PDF (260KB) ( 852 )   Save
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    【Objective】 In order to provide a reference for selection of reasonable species for eggplant grafted during winter planting, the contribution of root and scion to cold resistance of grafted eggplant seedlings was studied. 【Method】 Two rootstock eggplant cultivars, Daidaro (T), susceptible to cold and Hiranasu (C) is more resistant, were used to be grafted on each other. Based on this strategy, the physiology parameters including electrolyte leakage, MDA content and main osmotic adjustment substances in grafted seedling leaves before and after low temperature stress were studied. 【Result】 After low temperature treatment electrolyte leakage and MDA content in T-leaves of TC/TC, TC/C, TC/T was significantly higher than C-leaf in the same combination, but were observably lower than that of T/T, meanwhile, the content of soluble sugar, soluble protein and proline in T-leaves of combination of grafted seedlings was notably higher than that of T/T. All the results above indicate that both root and shoot of Hiranasu can enhance cold resistance of Daidaro in grafting experiment. Although TC/C had more resistance than TC/TC and TC/T which is the most sensitive to cold, all three of them were in the shade compared to C/C. So it can be concluded that both root and shoot of Daidaro had a negative effect on cold resistance of scion Hiranasu. 【Conclusion】 TC/C was more cold resistance than TC/T, so root had more contribution than shoot to cold resistance of grafted seedlings.

    Cloning and Sequence Analysis of the CP Gene of Citrus tatter leaf virus

    SONG Zhen,LIU Ke-hong,YANG Fang-yun,TANG Ke-zhi,LI Zhong-an,ZHOU Chang-yong
    Scientia Agricultura Sinica. 2009, 42(10):  3741-3748 .  doi:10.3864/j.issn.0578-1752.2009.10.0046
    Abstract ( 1053 )   PDF (379KB) ( 1057 )   Save
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    【Objective】 This study was conducted to investigate the molecular variability of the coat protein (CP) gene of Citrus tatter leaf virus (CTLV) isolates collected from China. 【Method】 The CP gene of 18 CTLV isolates from different geographical origins and citrus varieties were amplified by reverse transcription-polymerase chain reaction (RT-PCR). The DNA products were cloned and sequenced, and sequence analysis was conducted by DNAMAN. 【Result】 The CP gene of 18 CTLV isolates were all 714 nucleotides in length, and the putative CP contained 237 amino acids (aa). The identities of nucleotide and deduced amino acid sequences of CP gene among 18 isolates ranged from 88.5% to 99.9% and 91.1% to 99.6%, respectively. Corresponding to the mutation G289→A or C, A409→C, and G414→T on nucleotide sequences of CP gene, amino acids at positions 97, 137, and 138 of CP were different between isolates expressing mild symptoms and isolates expressing severe symptoms in the indicators. Q97 or K97, Q137, H138 were discovered in most mild isolates, whereas E97, R137 or K137, Q138 were shared by most severe isolates. Phylogenetic trees based on the aa sequences of the coat proteins showed that 18 CTLV isolates were divided into two clusters, and most of the mild isolates (4/6) belonged to theⅠgroup and most severe isolates (10/12) belonged to theⅡgroup. 【Conclusion】 The CP gene of CTLV was relatively conserved. The differences between mild and severe strains are very subtle, with three nucleotide positions (289, 409 and 414) appearing to determine the pathogenicity of virus.

    Construction and Analysis of Subtractive cDNA Library of Secondary Follicle in Anagen in Cashmere Goat
    XI Hai-yan,ZHOU Huan-min,ZHENG Yan
    Scientia Agricultura Sinica. 2009, 42(10):  3749-3754 .  doi:10.3864/j.issn.0578-1752.2009.10.0047
    Abstract ( 954 )   PDF (300KB) ( 658 )   Save
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    【Objective】 This study was carried out to construct suppression subtractive hybridization(SSH) library of secondary follicle in anagen and to search some candidate genes involved in cashmere development in Inner Mongolia cashmere goat. 【Method】 cDNA subtracted library was performed with cDNA from secondary follicle in anagen as the “tester” and cDNA from secondary follicle in telogen as the “driver”. The gene fragments were sequenced and analyzed by bioinformatics. 【Result】 The cDNA library was constructed successfully. Twenty positive clones were amplified by using nested PCR primer 1 and 2R, the size of inserts was 250-1 000 bp. Three hundred and fourty-two genes were obtained by DNA sequencing from 750 positive clones picked randomly. Their average length is 596 bp. After nucleotide blast homological analysis, 298 matched to known genes, 38 matched only to other ESTs in dbEST, and the remaining six showed no match to any ESTs or known genes. A total of 298 unique known genes were used to analyze the gene expression patterns. These were categorized into seven categories on the basis of gene function. They were divided into cell division (13), cell signaling/communication (49), cell structure/motility (52), cell/organism defense (21), metabolism (46), gene/protein expression (37), and unclassified (80). 【Conclusion】 The subtractive cDNA library of secondary follicles in anagen and telogen were obtained by SSH, and the information generated in this study had established a basis for screening candidate genes involved in cashmere development.