Scientia Agricultura Sinica

Wheat Photosynthetic and Temperature Production Efficiency Characters and Varieties Screening Based on Recombinant Inbred Lines Populations

DU Li-Feng, GAO Li-Feng, ZHOU Rong-Hua, SONG Yan-Xia, ZHAO Dong-Cheng, JIA Ji-Zeng, REN Zheng-Long

Scientia Agricultura Sinica. 2013, 46(8): 1523-1532. doi:10.3864/j.issn.0578-1752.2013.08.001

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【Objective】Study on the photosynthetic and temperature production efficiency (P&TPE) genetically and biologically will provide a theoretical basis for high-P&TPE wheat varieties breeding.【Method】The statistical methods of regression and clustering were applied to illustrate the relationship between P&TPE and main agronomic characters of RIL populations derived from a cross between Neixiang 188 and Yanzhan 1(NY). Association analysis was used to identify loci significantly related to P&TPE. Molecular marker-assisted selection was used in cultivating wheat new varieties of H-P&TPE. 【Result】Positive correlations were detected between P&TPE and spikes per plant, grain number per spike, average filling rate, the maximum filling rate of wheat, of which, spikes per plant had the highest effects on P&TPE. PPE and TPE were influenced greatly by the average and maximum filling rate, respectively. Eleven loci were identified to be significantly associated with P&TPE in at least two environments, of which, four loci (Xwmc167-2D, Xcwm23-2D, Xbarc218-3A and Xwmc326-3B) contributed more than 10% of the phenotypic variations. By molecular marker selection, five lines were screened from the 49 lines analyzed here which showed excellent performance in yield and P&TPE. 【Conclusion】There existed genetic loci conferring P&TPE in wheat genomes. Loci selection in populations with elite agronomic traits is an alternative approach to the low efficiency of MAS.

Genetic Analysis on Male Sterility of Thermo-Photo-Sensitive Male Sterile Line BNS in Wheat

ZHANG Bao-Lei, ZHANG Wei-Dong, GAO Qing-Rong, WANG Mao-Ting, LI Nan-Nan, ZHANG Yan-Yu, WANG Hui-Na, GAO Jian-Hua, ZHAO Lan-Fei, RU Zhen-Gang

Scientia Agricultura Sinica. 2013, 46(8): 1533-1542. doi:10.3864/j.issn.0578-1752.2013.08.002

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【Objective】 The objective of this research is to explore the genetic characteristics of new eco-genetic male sterile line BNS, to provide a theoretical guidance for breeding of wheat, and to lay a foundation for the further study on abortion mechanism of BNS. 【Method】 Reciprocal combinations between BNS and eight varieties ( lines ) were constructed to evaluate the cytoplasmic effects in BNS male sterility. Fertilities of F1 and F2 generations from cross of BNS/Shannong 055525 were analyzed in successive three years to obtain the optimal mode by applying the major gene plus polygene model of quantitive traits, and genetic parameters were also estimated. 【Result】 The fertility of BNS was controlled chiefly by nuclear genes, but cytoplasmic effects existed in some varieties (lines). The fertility of F2 populations showed a continuous distribution, and had obvious multi peak or skew phenomenon. The fittest genetic models was E_1, and that is the mode of two additive-dominant-epistasis major genes plus additive - dominant and additive-dominant gene. The major gene heritability was 72.5%-79.7%, and the major gene heritability was 4%-11.6%. The proportion of environmental variance was 8.8%-23.6% in phenotype variance. Male sterile gene expression in BNS was mainly regulated by the temperature, which leading to the changes of F2 seed setting rates and genetic parameters.【Conclusion】 The male fertility in BNS was controlled by two major genes plus polygene, and a certain amount of cytoplasmic effects was preliminarily found . The two major gene have more effects on the fertility, and their additive effect are much more than their dominant effects. The male sterile lines transformed or improved from BNS might be selected in early generations to enhance the efficiency of breeding.

Observation and Research on the Temporal and Spatial Distribution of Flowering and Flower Dropping of Soybean

ZHAO Shuang-Jin, TANG Xiao-Dong, ZHAO Xin, FENG Yan, ZHAO Cong-Cong, ZHANG Meng-Chen

Scientia Agricultura Sinica. 2013, 46(8): 1543-1554. doi:10.3864/j.issn.0578-1752.2013.08.003

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【Objective】 The whole progress of flowering and flower dropping of soybean were observed, and the characteristics and rules of different varieties were analyzed to provide an evidence for high yield breeding and cultivation. 【Method】 Under the conditions of field experiment, the flowering and flower dropping nodes of 72 individuals of six soybean varieties were observed daily (42 days). 【Result】 Flowering process showed a significant ladder of four stages: early florescence is short (about 3 days), less than 4 flowers per day, the flower amount at this stage is 5.73, 4.09% of the total, concentrated in the middle of plants. At full-bloom stage, the daily flowering 5-12 flowers, the duration is longer (12-18 days), the flowering amount of this stage is 108.75, 77.60% of total, the flowering section extends from the middle to the upper and lower ends, and middle-up. At slow florescence stage, the daily flowering 1-5 flowers, the duration is different among varieties, the flowering amount at this stage is 21.24, 15.16% of total, distributed in various nodes of the plant. At late florescence stage, the daily flowering less than 1 flower, until flowering end, the flowering amount is 4.42, scattered on the plants. Flower dropping generally occurs from early florescence 3 days later to late florescence or after late florescence, the flower dropping section throughout the flower section on the plants, the flower-dropping rate at the full flowering stage (36.97%) is less than at the initial flowering stage (51.76%), its slow flowering stage (65.54%), and its final flowering stage (77.96%). The peak of the abnormal abscission about 3-9 flowers daily is correlated with the rainy day and scant sunlight in that periods. In determinate podding habit varieties, the earliest flowering section is in the middle of the plants, and then extend to upper and lower sections, then extend to the bottom 3-7 days later and extend to the top 9-15 days later after flowering. In semi-determinate podding habit varieties, the earliest flowering section is in the middle and lower parts (7th-10th node), and then extend to upper and lower sections, then extend to the bottom 3-7 days and extend to the top 24-26 days later after initial flowering. 【Conclusion】 The flowering amount, flowering node, duration and abscission rate are all different in early florescence, full-bloom stage, slow florescence and late florescence of soybean.

Study on Genome Variations by Using SCoT Markers During Allopolyploidization of the Cultivated Peanut ×A. chacoensis

HE Liang-Qiong, XIONG Fa-Qian, ZHONG Rui-Chun, HAN Zhu-Qiang, LI Zhong, TANG Xiu-Mei, JIANG Jing, TANG Rong-Hua, HE Xin-Hua

Scientia Agricultura Sinica. 2013, 46(8): 1555-1563. doi:10.3864/j.issn.0578-1752.2013.08.004

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【Objective】Genome changes during allopolyploidization were studied to reveal preiliminarily the molecular mechanisms of Arachis polyploidization. 【Method】 A newly interspecific hybridization between tetraploid cultivated peanut Zhongkaihua No.4 and diploid wild one A. chacoensis to study the genomic varied time, types and frequency of the hybrid F1, early allopolyploidy generation (S0-S3) by start codon targeted polymorphism (SCoT) molecular marker technique. 【Result】 A total of 126 loci were produced including 117 polymorphic loci with a polymorphism rate of 92.86% by 18 SCoT primers, which showed that SCoT loci could reveal the relatively high genetic diversity among the Arachis collections. Compared with 109 parental fragments, 28, 30, 10,11 and 10 ones were lost and 9, 3, 10, 14 and 8 new fragments were produced in F1-S3 independently. Furthermore, in the lost parental bands, 16, 12, 7, 9 and 9 bands were from male. These results indicated that SCoT products began to alter as early as in F1 and the changes mainly included the loss, simultaneous appearance of parental bands and appearance of novel fragments, and the loss of male fragments was the major. 【Conclusion】The ATG translation start codon and the flanking region varied extremely in early period during Arachis allopolyploidization which speculated that their biological function might be related with polyploidy evolution and stability. As a new molecular marker technique, SCoT could provide a scientific basis in research on genomic variation during polyploidization in Arachis and other species because of its simplicity, effectiveness and practicality.

Research on Carbon Metabolism Characteristics of Transgenic Bt Rice

WANG Wei-Bo, CUI Hai-Rui, LU Mei-Zhen, SHU Qing-Yao, SHEN Sheng-Quan

Scientia Agricultura Sinica. 2013, 46(8): 1564-1570. doi:10.3864/j.issn.0578-1752.2013.08.005

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【Objective】The objective of this study was to evaluate the effect of transgene Cry1Ab on carbon metabolism characteristics in rice. 【Method】Transgenic Bt (Cry1Ab) rice KMD1 and its parent Xiushui 11, as well as three pairs of Bt and non-Bt rice near-isogenic lines which were derived from the progeny of crosses and continuous backcrosses between the KMD1 and Minghui 63, R3027 and 99Ya162, respectively, were field grown and measured for the content of chlorophyll and soluble sugar, the activity of RuBPCase, sucrose synthetase and sucrose phosphate synthase in leaves, the dry weight and organic carbon accumulation per plant at different developmental stages.【Result】At tillering stage, KMD 1 had significant lower contents of chlorophyll a, chlorophyll b and soluble sugar, lower activities of sucrose synthetase and sucrose phosphate synthase, and lower dry weight and organic carbon accumulation than those of Xiushui 11, but no significant differences between other 3 pairs of Bt and non-Bt rice lines were observed in all physiological indexes tested. At full heading stage, the activity of sucrose phosphate synthase of KMD1 was significantly higher than that of Xiushui 11, although its dry weight and organic carbon accumulation were significantly lower, while Bt R3027 showed a significant lower sucrose synthetase activity and Bt 99Ya162 showed significant higher soluble sugar content and sucrose phosphate synthase activity than the non-Bt type, respectively. At maturity stage, no significant differences were found in dry weight and organic carbon accumulation in four pairs of Bt and non-Bt rice lines. 【Conclusion】Significant change of most physiological indexes related to carbon metabolism in transgenic rice KMD 1 was resulted from somaclonal variations, while the effect of Cry1Ab gene on individual index in other 3 pairs of Bt and non-Bt rice near-isogenic lines was transitory and related to the genetic background of Bt gene incorporation.

Effects of Plant and Row Allocation on Population Light Environment and Lodging Resistance of Strip Sown Wheat in Drill

ZHENG Ting, CHEN Yi, FAN Gao-Qiong, LI Jin-Gang, LI Chao-Su, RONG Xiao-Jiao, LI Guo-Rui, YANG Wen-Yu, GUO Xiang

Scientia Agricultura Sinica. 2013, 46(8): 1571-1582. doi:10.3864/j.issn.0578-1752.2013.08.006

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【Objective】 The objective of this study was to screen an optimum plant and row space allocation pattern for mechanical strip sown wheat with high yield and lodging resistance in hilly regions of Sichuan.【Method】Two-factor split-plot design was applied in an experiment. The main-plot was two plant-type wheat cultivars, Chuannong 27 of compact-short and Mianmai 367 of intermediate type, and the sub-plot was seven plant and row allocation patterns with different numbers of rows and row spacing under 2 m wide strip type and 150×104 hm-2 planting density unchanged condition. The traditional double-three-zero strip planting pattern, planting five rows with row spacing of 20 cm+20 cm+20 cm+20 cm in a strip, as the CK. F3-1, F3-2 and F3-3 indicated planting three rows with row spacing of 30 cm+30 cm, 35 cm+35 cm and 40 cm+40 cm, respectively. F4-1, F4-2 and F4-3 indicated planting four rows with row spacing of 20 cm+20 cm+20 cm, 20 cm+30 cm+20 cm and 20 cm+40 cm+20 cm in a strip, respectively. The effects of reducing the number of planting rows and widening row spacing on light transmission rate, lodging resistance and yield were investigated, and the correlation of light transmission rate with lodging resistance was analyzed.【Result】Two wheat cultivars demonstrated consistently. Lodging resistance of edge row was significantly higher than that of inner row, and lodging resistance of inner row was positively correlated with light transmission rate. After reducing the number of planting rows from 5 to 4 or 3 and widening row spacing, light transmission rate was significantly improved, and lodging-resistance of edge row and inner row were enhanced, too, especial for the inner row. The related lodging resistance index of inner row indicated that plant height and height of gravity centre decreased, and the second basal culm internode became shorter and thicker, meanwhile, dry weight per stem, wall-thickness, mechanical strength, culm dry density, culm filling degree, the cellulose and lignin content increased, while lodging index decreased. Furthermore, the border advantage was cut down, and the expression of lodging resistance showed well in field test. Besides, spike number and yield increased. The edge row of CK had the best culm quality and the strongest lodging resistance, but the light transmission rate was the lowest, and culm quality of inner row showed serious deterioration, especial for the middle row. What’s more, CK demonstrated the strongest border advantage and the highest lodging odds in field test. In addition, the yield of CK was lower than that of F3-3 and F4-3, and was equivalent to that of F3-2 and F4-2.【Conclusion】Therefore, the allocation pattern, planting three rows with row spacing of 40 cm+40 cm or planting four rows with row spacing of 20 cm+40 cm+20 cm in a strip, is the optimum plant and row allocation pattern for mechanical sowing strip wheat with high yield and lodging resistance in hilly regions of Sichuan, and the allocation pattern, planting three rows with row spacing of 35 cm+35 cm or planting four rows with row spacing of 20 cm+30 cm+20 cm in a strip, is the second. On the whole, the above four patterns can replace the traditional double-three-zero strip planting pattern.

The Possible Effects of Global Warming on Cropping Systems in China Ⅷ—The Effects of Climate Change on Planting Boundaries of Different Winte-Spring Varieties of Winter Wheat in China

LI Ke-南, YANG Xiao-Guang, MU Chen-Ying, XU Hua-Jun, CHEN Fu

Scientia Agricultura Sinica. 2013, 46(8): 1583-1594. doi:10.3864/j.issn.0578-1752.2013.08.007

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【Objective】 Under the background of global climate change, the temperature in winter increased significantly from 1980s in China, how would this change affect the planting boundary of different winterness varieties of winter wheat in China? In order to answer this scientific question, the authors took the year 1981 as a time node and divided the interval 1951-2010 into two periods, then comparatively analyzed the change characteristics of north planting boundaries, south planting boundary and possible planting areas for strong winterness, winterness, weak winterness, and springness type winter wheat varieties in China due to increased temperature in winter during the latter period.【Method】 The northern and southern boundaries of different winter wheat types were defined according to the freezing damage index which restricted normal overwintering of winter wheat and the index which affects vernalization days of winter wheat. The changes of planting areas as well as coverage square meters of different winter wheat types were drawn with ArcGIS software. 【Result】 Compared with the period 1951-1980, in the period 1981-2010, the northern planting boundary of strong winterness winter wheat showed the most significant northward moving trend along Ningxia to Gansu and Hebei to Liaoning, with a spatial shift of 200 km and 100 km, respectively, and the southern planting boundary of strong winterness winter wheat showed a relatively more significant northward moving trend in the eastern region than the western region, with a spatial shift of 90 km along Jiangsu and Anhui zone, in addition, the possible planting area of strong winterness winter wheat in China increased by 36.24×104 km2 in total. The northern planting boundary of winterness winter wheat moved significantly along Shandong and Hebei, with a northward shift of 310 km, and the southern planting boundary of winterness winter wheat showed a significant westward moving trend along Bijie and Xishui stations in Guizhou, with a spatial shift of 95 km, also, the possible planting area of winterness winter wheat in China increased by 17.75×104 km2 in total. The northern planting boundary of weak winterness winter wheat showed a significant change trend around the common border of Anhui, Jiangsu, Henan and Shandong, with a northward shift of 120-370 km, yet the change trend in the western region was insignificant, the southern planting boundary of weak winterness winter wheat showed a slight northward moving trend, nevertheless, the possible planting area of weak winterness winter wheat in China increased by 15.70×104 km2 in total. The northern planting boundary of springness type winter wheat showed a significant moving trend along Jiangsu, Anhui and Henan, with a northward shift of 230 km, however, the change trend in the western region was insignificant, and the possible planting area of springness type winter wheat in China increased by 23.44×104 km2 in total. The north of North China was oriented by strong winterness winter wheat variety, South China was oriented by springness type winter wheat, there were relatively more possible planting winter wheat varieties in Henan, Shandong and Sichuan while winterness and weak winterness winter wheat were the oriented varieties.【Conclusion】 Due to the temperature in winter increasing obviously in China, compared to period 1951-1980, the planting boundaries of different winter wheat varieties in China moved northward significantly in period 1981-2010, and the northward moving trend of northern planting boundaries was more than that of southern planting boundaries, the possible planting area of winter wheat increased in all, especially, the strong winterness winter wheat showed the most significant moving trend in planting boundaries and possible planting area.

Characteristics of Adsorption and Desorption of Aluminum in Pectin as Influenced by Boron

LI Mei, YU Min-

Scientia Agricultura Sinica. 2013, 46(8): 1595-1602. doi:10.3864/j.issn.0578-1752.2013.08.008

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【Objective】The influence of boron on adsorption and desorption of aluminum in pectin were studied in order to probe the possible mechanism of boron in alleviating aluminum toxicity. 【Method】Isothermal adsorption method was conducted to study the influence of different concentrations of boron on the adsorption/desorption of aluminum in pectin under the conditions of pH 3.5 and 25℃.【Result】The total amount of aluminum adsorpted/desorpted in pectin significantly increased with the increase of pectin in equilibrium solution, but it significantly decreased in unit pectin. The amount of aluminum adsorpted/desorpted increased significantly with the increase of aluminum concentration as well. Boron pretreatment affected the characteristics of aluminum adsorption/desorption in pectin. Low concentration of boron less than 25 µmol•L-1 inhibited the adsorption/desorption of aluminum in pectin while the opposite was found at concentration higher than 25 µmol•L-1. When boron concentration was 100 µmol•L-1, the amount of aluminum adsorption in pectin was even more than that of the control without boron. But aluminum adsorption in pectin reduced when boron concentration was 200 µmol•L-1, suggesting that pectin aluminum adsorption reached saturation. The curves of aluminum adsorption could be well described by both Langmuir and Freundlich isotherm adsorption equations with determination correlation coefficient R2 over 0.97. The maximum adsorption capacity calculated by Langmuir decreased from 5.757 mg•g-1 to 0.160 mg•g-1 after adding boron, and the parameters of derived by Freundlich equation were 1.4702 and -0.1758, respectively, which indicated that the cross-linking of pectin RG-Ⅱ with boron affected strongly on aluminum adsorption in pectin. 【Conclusion】Boron could effectively inhibit aluminum adsorption in pectin at low concentration, while promote aluminum adsorption in pectin at high concentration.

StSte12 Expression Characteristics and Preliminary Screening of Its Regulatory Genes in Setosphaeria turcica

ZHANG Chang-Zhi, LI Po, GU Shou-Qin, GONG Xiao-Dong, YANG Yang, FAN Yu, TIAN Lan, ZHANG Xiao-Yu, HAN Jian-Min, DONG Jin-Gao

Scientia Agricultura Sinica. 2013, 46(8): 1603-1609. doi:10.3864/j.issn.0578-1752.2013.08.009

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【Objective】 The objective of this study is to locate StSte12 in genome, analyze its protein structure, confirmate its expression at different developmental stages, screen its regulatory genes which expressions were compared in Setosphaeria turcica.【Method】The location of StSte12 and its protein structures were analyzed using bioinformatics method. Its expression characteristics were detected by Western blot. The regulatory genes of StSte12 were screened according to ScSte12 of Saccharomyces cerevisiae. The binding motif of these genes was analyzed by bioinformatics. The expression level of these genes was compared between wild type strain and StSTE12 RNAi transformants using RT-PCR.【Result】The identity of StSTE12 was 105656, locating 1061747 to 1064127 of positive-strand of scaffold_13. StSte12 contained conserved domains (STE homeodomain and ZnF_C2H2 domain) and spatial structure of Ste12-like. In S. turcica appressorium period, the expression amount of StSTE12 was the largest. Some regulatory genes of StSte12 were screened, in which the binding motif combined with StSte12 was T\C GAAAC A\G. The expression level of StKAR5 in wild type strains was lager than that in StSTE12 RNAi transformants, the level of StBEM2, StBUD8, and StCHS7 were opposite to StKAR5, whereas that of StRAX2 had no obvious difference. 【Conclusion】 StSte12 has DNA binding domain and spatial structure belongs to Ste12-like Transcription factors. And it plays an important role in appressorium development of S. turcica. Some regulatory genes of StSte12 were screened, in which the binding motif was T\C GAAAC A\G, and which was regulated by StSte12.

Effects of Photoperiod and Temperature on the Post-Diapause Biology of Chrysoperla sinica (Tjeder) Adults in Different Overwintering Periods

CHEN Zhen-Zhen, LI Ming-Gui, GUO Ya-Nan, YIN Xiang-Chu, ZHANG Fan, XU Yong-Yu

Scientia Agricultura Sinica. 2013, 46(8): 1610-1618. doi:10.3864/j.issn.0578-1752.2013.08.010

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【Objective】The effects of photoperiod and temperature on the post-diapause development of Chrysoperla sinica adults and the relationship between diapause development and diapause intensity, diapause terminate, and post-diapuse development were studied. 【Method】 The pre-ovipositon, oviposition period, fecundity and longevity of overwintering adults of Ch. sinica were determined in November, January and March under three constant temperatures (18, 22, 28℃) and three different photoperiods (15D﹕9L, 13.5D﹕10.5L, 12L﹕12D). 【Result】Temperature had significant effects on pre-oviposition period (P＜0.001), oviposition period (P＜0.001) and longevity of female adults (P＜0.001) that shortened with the temperature increases. The fecundity had significant differences at different temperatures in November (P＜0.001) and the highest reproductivity was at 22℃ and the lowest at 18℃. However, no significant differences were found among temperatures in January (P=0.604) and March (P=0.745). Overwintering periods had distinct effects on the pre-oviposition period (P＜0.001), fecundity (P＞0.05) except oviposition periods (P＞0.05) and longevity of female adults (P＞0.05). The pre-oviposition periods and longevity of the adults became shorter with the overwintering time extending. The highest fecundity at 18℃ and 28℃ was in January, and the lowest in November, but at 22℃, the highest and lowest fecundity were in November and March, respectively. The photoperiods had no significant effects on pre-oviposition periods (P=0.240), oviposition periods (P=0.234) and female longevity (P=0.155) in the diapause termination process except fecundity (P＜0.001). The reduced light times could decrease significantly the fecundity of the adults in November (P=0.008) and March (P＜0.001), but no significant effects on the adults in January (P=0.735). Interactive analysis showed that temperatures and overwintering periods had significant effects on the pre-oviposition period (P=0.016) and fecundity (P＜0.001), but no significant effects on oviposition periods (P=0.321) and female longevity (P=0.583). Photoperiod and overwintering periods had no significant interactive effects on the pre-oviposition period (P=0.664), oviposition periods (P=0.074) and female longevity (P=0.052), but significant effects on fecundity (P=0.031).【Conclusion】Temperature has significant effects on the post-diapause biology of Ch. sinica adults. The overvintering periods have significant effects on pre-oviposition period and fecundity, this indicates that the green lacewing has different diapause intensities in different months and would influence the fecundity because of the energy consumption during the process of diapause termination. The photoperiod has no significant effects on the pre-oviposition period, oviposition periods and female longevity, but significant effects on fecundity, the results reflect that light quality can affect egg production. Temperature and overvintering periods have more significant effects on the post-diapause development of Ch. sinica than the photoperiod.

Effects of Different Land Use Types on Soil Physical Properties and Their Distributions in Farming-Pastoral Ecotone of Northern Shaanxi

BAI Yi-Ru, WANG You-Qi, ZHAN Xiu-Li

Scientia Agricultura Sinica. 2013, 46(8): 1619-1627. doi:10.3864/j.issn.0578-1752.2013.08.011

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【Objective】The land use types can lead to the differences of land cover conditions which influence the changes of soil properties and cause different environmental effects. 【Method】 Field sampling and lab analysis were combined to identify the soil physical properties and their profile distributional characteristics for millet, almond, alfalfa and S. bungeana in farming-pastoral ecotone of northern Shaanxi in 2010. 【Result】 The results showed that the differences in soil bulk density, capillary porosity and water holding capacity were significant in different land use types, while there were no significant differences in soil particle composition and saturated hydraulic conductivity. The soil fractal dimension values under different land use types decreased in the following order: alfalfa (2.490) ＞almond (2.485) ＞S. bungeana (2.459) ＞millet (2.445), which showed that soil particles were coarsened seriously on the land of millet. The field water capacity decreased in the following order: alfalfa (18.02%) ＞almond (17.49%) ＞S. bungeana (17.19%) ＞millet (14.27%), which indicated that water permeability, aeration and water holding capacity of soil on woodland and grassland were compatible. The distribution characteristics of soil physical properties were obviously different in profile. The soil physical properties changed greatly and showed volatility in the profile of millet land. The bulk density increased, while the soil capillary porosity and saturated hydraulic conductivity decreased gradually on the land of alfalfa, almond and S. bungeana as the soil depth increased. 【Conclusion】The research demonstrated that the farming cultivation could worsen the soil porosity and destroy the soil structure. Consequently, the conversion of cropland to forest and grassland project should be strongly carried out, and almond, alfalfa and Stipa bungeana suitable for the local ecological environment should be planted to improve the soil quality of the region.

Effects of Long-Term Different Fertilizations on the Fate of Residual Fertilizer N in a Wheat-Maize Rotation System

ZHAO Wei, LIANG Bin, YANG Xue-Yun, ZHOU Jian-Bin

Scientia Agricultura Sinica. 2013, 46(8): 1628-1634. doi:10.3864/j.issn.0578-1752.2013.08.012

Abstract ( 731 )

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【Objective】The fate of residual fertilizer N was studied in soils with different fertilities. 【Method】The aim of this experiment was to study the fates of residual fertilizer N after three consecutive growing seasons in the soils with a history of long-term different fertilizations (including no fertilizer (No-F soil), inorganic NPK fertilizer (NPK soil), and cattle manure plus inorganic NPK (MNPK soil)) using the 15N microplot method. 【Result】The total urea-15N use efficiency in the NPK and MNPK soils (57% and 65%, respectively) during the three growing seasons was significantly (P＜0.05) higher than that in the No-F soil (28%). The uptake rate of the residual urea-15N by the crops growing in the subsequent two seasons were 17% for the No-F soil, 15% for the NPK soil, and 8% for the MNPK soil. And 7% of the residual urea-15N was taken up by the third crops in the No-F soil, 2% in the NPK soil and 3% in the MNPK soil. The total urea-15N recovery rate (urea-15N uptake by the three crops and urea-15N residual in the 0-100 cm soil layers) were 50% in the No-F soil, 77% in the NPK soil and 84% in the MNPK soil. And the total loss rate of urea-15N were 50% in the No-F soil, 23% in the NPK soil and 16% in the MNPK soil after the three growing seasons. The residual rate of the urea-15N in the 0-100 cm soil layers was about 20% in all soils after the three growing season, mainly in the 0-20 cm soil layers.【Conclusion】 The conclusion is that the residual fertilizer N has high availability for the subsequent crops, and the long-term combined application of cattle manure and inorganic NPK can increase fertilizer N use efficiency and decrease its loss.

Simulation and Analysis of Nitrogen Loss, Water and Nitrogen Use Efficiencies of Greenhouse Cucumber Under Different Water and Fertilizer Managements

SUN Yuan, HU Ke-Lin, QIU Jian-Jun, JIANG Li-Hua, XU Yu

Scientia Agricultura Sinica. 2013, 46(8): 1635-1645. doi:10.3864/j.issn.0578-1752.2013.08.013

Abstract ( 711 )

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【Objective】 The study was to quantify the nitrogen (N) loss, and water and nitrogen use efficiencies (WUE, NUE) of greenhouse vegetable under different water and fertilizer managements using the EU-Rotate_N model, and to provide some basic suggestions on greenhouse vegetable cultivation. 【Method】 A field experiment was conducted in a cucumber greenhouse located in Shouguang city, Shandong Province, from 2010 to 2011. Four different water and fertilizer practices were designed, including no fertilizer + furrow irrigation (CK), conventional fertilization +furrow irrigation (farmer’s practice, FP), optimal fertilization + furrow irrigation (OPT), and conventional fertilization + drip irrigation (RI). The EU-Rotate_N model was used to simulate the yield, nitrate leaching, and gaseous N loss, during spring-summer (SS) and autumn-winter( AW) seasons. WUEs and NUEs of all treatments were also calculated. 【Result】 The results showed that irrigation water use efficiencies under drip irrigation (RI) increased by over twice compared with furrow irrigation treatments (CK, FP and OPT) with saving 60% of irrigation input in both growing seasons. For all fertilization treatments, the amount of gaseous N loss accounted for 16%-19% and 6%-11% of total N input in SS and AW seasons, respectively. The amounts of nitrate leaching accounted for 14%-57% and 20%-55% of total N application in SS and AW seasons, respectively. Compared with FP treatment, the amount of nitrate leaching in OPT and RI treatments reduced by 19%-31% and 63%-76%, respectively. Meanwhile, the NUE in OPT treatment increased by 3% and 7%, and the NUE in RI treatment increased by 41% and 44% in SS and AW seasons, respectively. 【Conclusion】 Nitrate leaching is a major component of N loss in greenhouse vegetable production in Shouguang County. Both optimal fertilization and drip irrigation practices could reduce the nitrate leaching and increase the NUE.

Effects of NaCl Stress on Seed Germination and DNA Methylation Status Detected by MSAP Analysis in Cucumber

HUANG Yun-Yu, ZHANG Hai-Jun, XING Yan-Xia, QI Yan, SUN Qian-Qian, ZHOU Chun-Lei, ZHAO Bing, GUO Yang-Dong

Scientia Agricultura Sinica. 2013, 46(8): 1646-1656. doi:10.3864/j.issn.0578-1752.2013.08.014

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【Objective】The objectives of this study were to analyze the cytosine methylation dynamics during cucumber seed germination and to assess the effect of NaCl treatment on seed germination and DNA methylation status. Moreover, the role of DNA methylation on seed germination and salt stress adaptation was studied at the same time.【Method】Methylation sensitive amplified polymorphism (MSAP) method was used to analyze the level and variation patterns of DNA methylation at different times (0, 1, 2, 4, 6, 8 d) during cucumber seed germination in different NaCl treatments (0, 150, 200 mmol•L-1).【Result】MSAP results showed that the methylation level in cucumber dry seeds was 15.25%, mainly in full methylation pattern. The level of global DNA methylation decreased during the whole germination process, with a slight increase at the early stages (1-2 d). NaCl treatment aggravated these changes, and made the final level of DNA methylation lower. Hypermethylation and hypomethylation happened simultaneously, and predominated at different germination stages. Among different change types, CG/CHG (H=A, T, C) simultaneous changes were the dominating types. DNA methylation changes were detected in both coding region and non-coding region on the cucumber genome.【Conclusion】DNA methylation changes during seed germination is a complex process. Methylation and demethylation happened at the same time in a temporal-spatial specific way. NaCl treatment reduced the cucumber genome stability and inhibited the seed germination.

Evaluation System Established for Fresh Apple Juice Quality

NIE Ji-Yun, WU Yong-Long, LI Hai-Fei, WANG Kun, XU Guo-Feng, YAN Zhen, WU Xi

Scientia Agricultura Sinica. 2013, 46(8): 1657-1667. doi:10.3864/j.issn.0578-1752.2013.08.015

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【Objective】 The aim of this study was to establish an scientific evaluation system for fresh apple juice quality. 【Method】 Sixteen indices of fresh apple juice quality were determined. Evaluation indices of fresh apple juice quality were screened by factor analysis and graded by probability distribution, and their weight was decided by analytic hierarchy process. Discrimination functions were established for fresh apple juice quality by K-means cluster and discriminant analysis. 【Result】 Indices of fresh apple juice quality (flavor not included) dispersed with coefficient of variation from 2.0% to 112.2%. From 16 indices, 7 indices were chosen as evaluation indices for fresh apple juice quality, including flavor, a value, b value, transmittance at 625 nm, turbidity, total soluble solid, and total soluble solid/titratable acidity. Both the grading standard and the scoring standard of above 7 indices were established. Five discrimination functions were established for fresh apple juice quality, which had satisfactory recognition accuracy up to 98.89% (modeling samples) and 100% (test samples). 【Conclusion】 Fresh apple juice quality can be evaluated by 7 indices, including flavor, a value, b value, transmittance at 625 nm, turbidity, total soluble solid, and total soluble solid/titratable acidity. The grading standard and the scoring standard of these 7 indices set up a scientific foundation for evaluation of fresh apple juice quality. The established discrimination functions are very effective in discriminating fresh apple juice quality.

Fluoride Accumulation and Its Subcellular Distribution in Three Tea Plants

CAI Hui-Mei, PENG Chuan-Yi, LI Cheng-Lin, GAO Zhu, HOU Ru-Yan, WAN Xiao-Chun

Scientia Agricultura Sinica. 2013, 46(8): 1668-1675. doi:10.3864/j.issn.0578-1752.2013.08.016

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【Objective】The objective of this study was to give some experimental basis for studying the absorption, accumulation of F and the mechanism of detoxification.【Method】Solution culture and differential centrifugation were conducted to study the characteristics of F accumulation and its subcellular distribution of Pingyangtezao, Wuniuzao and Fuzao No.2, with comparison of the differences between varieties.【Result】F concentration in roots, stems and leaves of three kinds of tea plants increased with F treatment, under 0-10 mg•L-1 F treatments, the ground part of fluoride content was greater than the root(Translation correlation ＞1) and 56% of F was distributed in above-ground part. Except for the control, most of the fluoride was distributed in cell wall (76.84%-91.58%) and soluble fraction (53.24%-80.35%) of leaf and root. F treatment generally increased the percentage of cell wall and soluble fraction, but decreased that of cell organelle. In F treatment, the change tendency of F concentration of subcellular fractions of leaf of Pingyangtezao and Wuniuzao ranked in the following order: cell wall＞ soluble fraction＞ cell organelle, while soluble fraction＞cell wall＞ cell organelle for Fuzao No.2 . In low F (0-5 mg•L-1 ) treatment , the change tendency of F concentration of subcellular fractions of root of three kinds of tea plants ranked in the following order: cell organelle≥cell wall＞soluble fraction, while cell wall＞soluble fraction≥cell organelle in high F (10-50 mg•L-1 ) treatment.【Conclusion】Translation correlation , percentage of F for above-ground part and F subcellular distribution in root and leaf suggested tea plant was of strong upward transportation capacity. The subcellular distribution of F in leaves and roots were related with tea species and solution F treatment, and so did the correlation between the concentration of T-F and subcellular fractions in leaf, with some differences, but no in root. Cell wall and soluble fraction were the main storage sites in high F treatment, suggesting cell wall binding and vacuolar compartmentalization may be main reasons for tea plant to detoxify fluoride toxicity.

Changes of the Metabolism of Starch and Sucrose Content in Chestnuts at Different Degrees of Calcification

GU Cai-Qin, LAI Ya-Ping, ZHU Dong-Xue, LIU Peng, ZENG Qing-Zhu

Scientia Agricultura Sinica. 2013, 46(8): 1676-1681. doi:10.3864/j.issn.0578-1752.2013.08.017

Abstract ( 503 )

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【Objective】The metabolism of starch and changes in sucrose in chestnuts in relation to calcification were studied in order to investigate the mechanism of calcification occurrence.【Method】The degrees of calcification in chestnuts stored at room temperature and RH 60%-70% for some days was divided into five grads 0%, 30%, 60%, 90%, and 100% and the moisture content, activities of amylase, content of starch and sucrose were determined. 【Result】With the increase of calcification, the contents of moisture and starch decreased obviously, the relationship between calcification in chestnut and the contents of moisture and starch was a negative correlation (r＝-0.946 and r＝-0.950). The activities of α-amylase and β-amylase increased firstly and then decreased, but the content of sucrose increased gradually.【Conclusion】The occurrence of calcification in chestnut is related to changes of crystalline, thermal and textural characteristics of starch which resulted by degradation of starch led by increase of amylase activities induced by loss of moisture , and no correlation with the content of sucrose.

Simultaneous Detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae Using Multiplex PCR

WEI Shuang, XIAN Yu-Yin, ZHAO Hui, WU Xi-Yang

Scientia Agricultura Sinica. 2013, 46(8): 1682-1686. doi:10.3864/j.issn.0578-1752.2013.08.018

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【Objective】The objective of this study is to develop a multiplex PCR assay that can simultaneously detect Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae in the presence of an internal amplification control (IAC).【Method】Species-specific PCR primers were designed based on gyrB gene for Vibrio alginolyticus, collagenase gene for Vibrio parahaemolyticus, vvhA gene for Vibrio vulnificus and ompW gene for Vibrio cholerae, 16S rRNA gene of bacteria as IAC primers was used to indicate false-negative results. Multiple PCR method was developed after optimization reaction condition. 【Result】The multiple PCR method was proved to be rapid, high-throughput, sensitive and specific and the existence of IAC could successfully eliminate false-negative results. The multiplex PCR was validated with 69 suspicious Vibrio strains and the results were consistent with the physiological and biochemical experiments.【Conclusion】The multiple PCR method is specific, stable and reliable for the detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae.

Screening and Analysis of Dipeptidyl Peptidase Ⅳ from Microbial Metagenomic Library in the Rumen of Dairy Cows

ZHAO Jing-Wen, WANG Jia-Qi, ZHAO Sheng-Guo, SUN Peng, BU Deng-Pan, HU Xiao-Li, LU Yu-Fei, WANG Dan-Dan, JIN Di

Scientia Agricultura Sinica. 2013, 46(8): 1687-1693. doi:10.3864/j.issn.0578-1752.2013.08.019

Abstract ( 633 )

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【Objective】The purpose of this study was to reveal the characteristics of dipeptidyl peptidase Ⅳ (DPP-Ⅳ) sequences and enzymatic activity during the process of protein degradation in the rumen of dairy cows.【Method】The dpp-Ⅳ degenerate primers were used to screen rumen microbial Fosmid library including 17664 clones. The plasmids were extracted from the positive clones and digested by Hind Ⅲ. The dpp-Ⅳ sequences were obtained by PCR, cloning and sequencing. The Fosmid end sequences and DPP-Ⅳ sequences of the positive clones were analyzed by BLASTX and BLASTP, respectively. The peptidase activity from the positive clones was measured using Gly-Pro-pNA as a substrate.【Result】Ten positive clones named DP1-DP10 containing dpp-Ⅳ fragment were obtained. 78% of the Fosmid end sequences could match with the known genes (similarity 44%-94%). DPP-Ⅳ sequences contained N-conservative region (DWVYEEE) and C-catalytic domain (GWSYGG). DPP-Ⅳ sequences matched to Cyclobacterium marinum (43%), Capnocytophaga sp. (63%), Prevotella ruminicola 23 (66%) and Solitalea canadensis (50%). The activity of DP7 peptidase is the highest (6.88 U•mg-1).【Conclusion】Ten positive clones obtained from rumen microbial Fosmid library had different sequence characteristics and peptidase activity. This study implies that the substitution of some DPP-Ⅳ AA may affect the enzyme activity.

Advances in Study on Bio-Energy Utilization of Stem Cell Wall Components in Alfalfa (Medicago sativa L.)

WANG Xiao-Juan, ZHANG Shu-Zhen, LIN Shuang-Shuang, DENG Zhi-Gang, JIN Liang

Scientia Agricultura Sinica. 2013, 46(8): 1694-1705. doi:10.3864/j.issn.0578-1752.2013.08.020

Abstract ( 545 )

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Alfalfa (Medicago sativa L.) has been recognized as one of the potential biofuel crops with high biomass and wild adaptability. Most importantly, the stems of alfalfa would be processed to produce ethanol and the leaves could be served separately as a livestock feed. In this paper, the recent advances of the studies and utilization of alfalfa as energy plant were reviewed, which including germplasm evaluations of biofuel traits, development of stem cell wall, biosynthetic pathways and genetic control of cell wall components (cellulose, hemi-cellulose and lignin), and the effects of cultivation management system to ethanol production. The development of stem cell wall and genetic regulation of lignin in alfalfa were discussed. Furthermore, the perspective utilization of alfalfa as biofuel crop was also proposed.

Molecular Cloning and Expression Profiles Analysis of Chemosensory Protein Genes Family in the Chinese Honeybee (Apis cerana cerana)

NI Cui-Xia, ZHANG Lin-Ya, LI Hong-Liang, SHANG Han-Wu

Scientia Agricultura Sinica. 2013, 46(8): 1706-1715. doi:10.3864/j.issn.0578-1752.2013.08.021

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【Objective】 The objective of this study is to clone the whole genes family of chemosensory protein in the Chinese honeybee (Apis cerana cerana) (Ac-CSPs) and analyze their expression of mRNA in different tissues and different development time-points. This may provide a fundamental evidence for the future study of the physiological function of Ac-CSPs. 【Method】 Six full-length of CSP genes of A. cerana cerana was cloned by using reverse transcription-polymerase chain reaction (RT-PCR). The expression patterns at development phases and in different sensory organs of the six Ac-CSPs were analyzed by real-time PCR. 【Result】 Four full-length of Ac-CSP genes (Ac-CSP2, Ac-CSP4, Ac-CSP5 and Ac-CSP6) were cloned in which Ac-CSP2 was 354 bp in full length that encoded a polypeptide containing 117 amino acids with prediction of MW (molecular weight) of 13.01 kD and pI of 8.86, respectively. Ac-CSP4 was 387 bp in full length that encoded a polypeptide containing 128 amino acids with prediction of MW of 14.61 kD and pI of 4.53, respectively. Ac-CSP5 was 315 bp in full length that encoded a polypeptide containing 104 amino acids with prediction of MW of 12.46 kD and pI of 9.60, respectively. Ac-CSP6 was 378 bp in full length that encoded a polypeptide containing 125 amino acids with prediction of MW of 14.63 kD and pI of 8.71, respectively, and all of the genes with four conserved cysteine residues. The six deduced Ac-CSPs sequences showed high sequence similarity (95%-99%) as compared with CSPs (Am-CSPs) sequences of A. mellifera. The genes of Ac-CSP1, 2, 3, 4 were not expressed in eggs, larvae and pupae, with the Ac-CSP1, 2, 4 expressed at highest level in antennae and Ac-CSP3 was strongly expressed in wings. The expression of Ac-CSP5 was not significantly different at all developmental stages, while Ac-CSP6 only highly expressed in the pupae. 【Conclusion】 The alignment is very similar of Ac-CSPs and Am-CSPs and the expression patterns are similar but there is a part of difference in this research, which provides useful information for the study of function of CSPs in the future.

Recent Advances in Research on Bovine Oocyte Maturation Technology in vitro

JIA Zhen-Wei, TIAN Jian-Hui, AN Lei, ZHANG Jia-Xin

Scientia Agricultura Sinica. 2013, 46(8): 1716-1724. doi:10.3864/j.issn.0578-1752.2013.08.022

Abstract ( 520 )

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Bovine oocyte in vitro maturation (IVM) is an important reproductive technology, however, as these oocytes were maturated in vitro that is deficient in growing process and some crucial event in vivo during antral follicular development prior to the LH surge, which interfere cytoplasmic maturation, thus resulting in reduced developmental capacity compared to in vivo maturated counterparts. Oocyte maturation is a complex process that involves events of nuclear, cytoplasmic and molecular maturation. Bovine oocyte maturation during in vitro culture is affected by many factors including source of oocyte, bidirectional communication between the oocyte and its surrounding cumulus cells and maturation environment. Up to date, to improve the developmental competence of bovine oocytes in vitro, some new technologies, for example, attenuating spontaneous oocyte maturation, improving oocyte maturation using exogenous oocyte-secreted growth factors and simulated physiological oocyte maturation have been developed for enhancing bovine oocytes maturation in vitro by simulating in vivo oocytes growing environment. This paper reviewed the maturation processes of bovine oocytes, the factors affecting oocyte maturation in vitro and the latest approach to improve the maturation rate of bovine oocytes in vitro.

Cloning and Bioinformatics Analysis of Full-Length cDNA Sequence of YAP1 Gene in Sheep

SUN Wei, LI Da, SU Rui, MA Yue-Hui, GUAN Wei-Jun, ZHANG You-Fa, CHEN Ling, WU Wen-Zhong, ZHOU Hong

Scientia Agricultura Sinica. 2013, 46(8): 1725-1735. doi:10.3864/j.issn.0578-1752.2013.08.023

Abstract ( 547 )

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【Objective】 There was no report on YAP1 gene and its protein in the past days, this study was carried out in order to clone the YAP1 cDNA in sheep and provide a basic foundation for the study of YAP1 genetic characters , as well as analyze its tissue different level expression patterns. 【Method】 Hu sheep was regarded as the research object, and the full-length cDNA sequence of the YAP1 gene was acquired from the longissimus dorsi muscle by RT-PCR and RACE, and the sequence was analyzed in depth by bioinformatics method. 【Result】 The full-length cDNA sequence of the YAP1 gene was successfully obtained from Hu sheep longissimus dorsi muscle. Bioinformatics technology was successfully used to analyze the YAP1 homology among different species, transmembrane region of YAP1 sequence, subcellular localization, hydrophilic, potential signal peptide cleavage sites, glycosylation sites, phosphorylationlocus, product function prediction analyisis, YAP1 secondary structure and teriary structure.【Conclusion】The full-length of YAP1 gene was 1 712bp, including 1 212bp coding sequence encoding 403 amino acids. The homology analysis found that the nucleotide sequence and amino acid sequence of sheep YAP1 gene share the highest 78.77% and 92.51% identity with the Cricetulus YAP1 while there was no large difference between Human and Chimpanzee. The amino acid sequence analysis revealed that the YAP1 gene of sheep encoded water-soluble protein and its relative molecular weight was 44079.0 Da, isoelectric point was 4.91, was a kind of hydrophilic non-transmembrane protein. Subcellular localization of YAP1 was in the mucleus, and it did not belong to the secreted protein. The YAP1 protein contained 33 phosphorylation sites, 7 glycosylation sites and 2 WW domain. The secondary structure of YAP1 was mainly composed of random coil. The tertiary structure of domain area of YAP1 protein showed a forniciform helix structure. Forecast YAP1 had a key role in regulatory functions process. The above studies have laid an information foundation for further study of the YAP1 gene function in the sheep muscle development process.

二斑叶螨乙酰辅酶A羧化酶基因全长cDNA克隆及其生物信息学分析

LU Juan-Juan,WANG Jin-Jun, GAO Xin-Ju, SHEN Hui-Min

Scientia Agricultura Sinica. 2013, 46(8): 1736-1744. doi:10.3864/j.issn.0578-1752.2013.08.024

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【Objective】The objective of this study is to clone full-length cDNA sequence of the Acetyl-CoA carboxylase (ACCase) gene from Tetranychus urticae and analyze its expression in different strains. 【Method】 The full length cDNA of ACCase gene from T. urticae was cloned by using RT-PCR, and the bioinformatics software was employed to analyze the characteristics of the coded protein. Expression profiles of ACCase gene between susceptible strain (S) and spirodiclofen resistant strain (Sp-R) of T. urticae were performed by the quantitative real-time PCR. 【Result】The full-length cDNA of ACCase gene (GenBank accessin number JX424763) contained 7 068 bp encoding 2 235 amino acids. The putative protein of ACCase gene showed predicted molecular weight of 266.35 kD with a theoretical pI of 6.38. The ACCase contained three function domains, including the biotin carboxylase (BC), the biotin carboxyl carrier protein (BCCP) and the carboxyltransferase (CT). The phylogenetic tree of nine ACCase amino acid sequences from different species were constructed by MEGA5.0．The results showed a close relationship with Pediculus humanus corporis. Quantitative real-time PCR showed that the transcripts of ACCase gene were 2-fold higher in spirodiclofen (Sp-R) resistant strain compared with susceptible strain.【Conclusion】The ACCase gene was cloned from T. urticae by using the RT-PCR. The higher expression of ACCase gene was probably related with the development of resistance to spirodiclofen, which may provide a basis for the management of pesticide resistance of T. urticae.

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