Scientia Agricultura Sinica

Variations of LOX and PPO Activities and Carotenoid Content as Well as Their Influence on Whole Flour Color in Common Wheat

ZHENG Wen-Yin, WANG Fan, SI Hong-Qi, ZHANG Wen-Ming, YAO Da-Nian

Scientia Agricultura Sinica. 2013, 46(6): 1087-1094. doi:10.3864/j.issn.0578-1752.2013.06.001

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【Objective】The aim of this study is to provide a theoretical basis for improving flour color and related traits in breeding practice. LOX and PPO activities and carotenoids concerntration of fresh and stored whole flours were determined to study their variation, analyze the effects of these traits on the whole flour color by correlation coefficient, and evaluate the influence of storage time on flour color.【Method】The LOX and PPO activities and carotenoids concerntration were determined spectrophotometrically in 63 wheat varieties. The whole flour color was evaluated by Color difference meter.【Result】There were significant or very significant variations of LOX and PPO activities and carotenoids concerntration as well as other color index in both fresh and stored whole flours. During the storage, the LOX and PPO activities reduced quickly, but the carotenoids concentration remained steadily. For fresh whole flour, the whiteness and brightness had a significant negative correlation with PPO activity, while yellowness showed a significant positive correlation with carotenoids concerntration. LOX activity was only significantly negatively correlated with carotenoid content whereas no significant relationships with the whole flour whiteness, brightness and yellowness were found. After storage for 3 months, the whole flour whiteness and yellowness were only significantly negatively correlated with PPO activity.【Conclusion】To improve the flour color, the genetic control of PPO activity and carotenoids concerntration is essential. Storing whole flour is an impracticable method as it cannot result in the increase of whole flour whiteness and brightness, and the reduction of yellowness in wide range. The genetic breeding would be an important way to improve flour whiteness or yellowness with high nutritional value.

Effects of High Molecular Weight Glutenin Subunits on Wheat Quality by Aroona and Its Near-Isogenic Lines

JIN Hui, HE Zhong-Hu, LI Gen-Ying, MU Pei-Yuan, FAN Zhe-Ru, XIA Xian-Chun, ZHANG Yan

Scientia Agricultura Sinica. 2013, 46(6): 1095-1103. doi:10.3864/j.issn.0578-1752.2013.06.002

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【Objective】The effects of high molecular weight glutenin subunits (HMW-GS) on dough properties, quantity of gluten protein fractions, and bread-making quality were determined under the same genetic background of low molecular weight glutenin subunits (Glu-A3c, Glu-B3b, Glu-D3c). 【Method】Near-isogenic lines (NILs) developed from Australian cultivar Aroona planted in Urumqi and Shihezi in Xijiang during the 2010 cropping season were used to investigate Farinograph, Extensograph, quantity of gluten protein fractions, and bread-making quality. 【Result】The effect of HMW-GS on extensibility was not significant. For dough strength, different HMW-GS loci were ranked as Glu-D1＞Glu-B1＞Glu-A1 and the subunit pairs 7+9, 17+18, and 5+10 were correlated with superior dough strength. The line with 1, 7+9, 5+10 performed the best dough strength while those with 2*, 7+9, 2+12 and 1, 7+9, 2.2+12 were associated with superior extensibility. For the percent of SDS-unextractable glutenin polymeric protein (%UPP), different HMW-GS loci were ranked as Glu-D1＞Glu-B1＞Glu-A1 and the subunit pairs 7+9, 17+18, and 5+10 exhibited the highest %UPP. The line with 1, 7+9, 5+10 was associated with the highest %UPP. For bread-making quality, different HMW-GS loci were ranked as Glu-D1＞Glu-A1＞Glu-B1 and the subunits 1, 2*, 2+12, and 5+10 showed the higher total score. The line with 1, 7+9, 2+12 had the highest total score, 1, 7+9, 5+10 showed the second highest total score, and null, 7+9, 2+12 displayed the lowest total score.【Conclusion】The effects of HMW-GS on dough strength, quantity of gluten protein fractions, and bread-making quality were significant under the same genetic background of LMW-GS (Glu-A3c, Glu-B3b, Glu-D3c); Subunits 1, 2*, 7+9, 17+18, and 5+10 were correlated with superior wheat quality and the line with 1, 7+9, 5+10 is ideal for improving bread-making quality.

Down-Regulation of the Expression of Grain ppo Genes to Improve Wheat Dough Whiteness by RNA Interference

SUN Zhong-Xia, YANG Feng-Ping, ZHANG Ting, SUI Xiao-Yan, LIANG Rong-Qi, LIUQing , ZHANG Xiao-Dong, LI Bao-Yun

Scientia Agricultura Sinica. 2013, 46(6): 1104-1113. doi:10.3864/j.issn.0578-1752.2013.06.003

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【Objective】 To develop the transgenic wheat germplasm with low flour PPO activity and high whiteness by RNA interference (RNAi) to specially down-regulate the expression of grain ppo genes. 【Method】 The RNAi vector pBAC47P-ppoIR, aimed at silencing wheat grain ppo genes, was transformed into the callus from Zhongyou9507 immature embryo by biolistic bombardment, and the transgenic plants were obtained after resistance selection, differentiation and regeneration, and then screened by PCR, Southern blotting, half-quantitative RT-PCR, northern blotting and enzyme activity analysis. 【Result】 A total of 27 transgenic T0 plants were obtained. Twelve transgenic lines of T1 generation confirmed by PCR and genomic Southern blotting showed successful integration of RNAi construct into the wheat genome. The expression of grain ppo genes of 20 T2 plants was significantly lower than that of control. Enzyme activity analysis of T2 grains at filling stage showed that the PPO activity of 6 lines reduced obviously. Dough whiteness analysis of T4 grains showed that the whiteness of 5 lines increased clearly and 2 lines improved significantly.【Conclusion】The RNAi vector pBAC47P-ppoIR could down-reglulate the expression of grain ppo genes, reduce the PPO activity, and increase the dough whiteness significantly, so this study will provide wheat breeder with breeding material for whiteness and quality improvement.

Evolution of Agronomic and Quality Traits of Wheat Cultivars Released in Shandong Province Recently

SONG Jian-Min, DAI Shuang, LI Hao-Sheng, CHENG Dun-Gong, LIU Ai-Feng, CAO Xin-You, LIU Jian-Jun, ZHAO Zhen-Dong

Scientia Agricultura Sinica. 2013, 46(6): 1114-1126. doi:10.3864/j.issn.0578-1752.2013.06.004

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【Objective】In order to provide a theoretical basis for further improvement of yield potential and processing quality, agronomic and quality traits of 55 wheat cultivars released in Shandong province recently were investigated.【Method】 Eighteen agronomic and quality traits of wheat cultivars released in Shandong province from 1999 to 2010 were analyzed, and their evolution and correlation were also discussed. 【Result】The yields of wheat cultivars released recently in Shandong province increased annually by 61.65 kg•hm-2，though main yield components such as spike number per unit area and kernels per spike changed insignificantly. The growth duration decreased annually by 0.57 d, and plant height and maximum tillers also decreased annually, whereas fertile tiller increased. Compared to the cultivars with lower yield, the cultivars with higher yield had more spikes per unit area and fertile tiller, suggesting that it may be an effective measure to increase yield by improving fertile spikes. Shandong wheat cultivars released recently had relatively high protein and wet gluten content, low Zeleny sedimentation volume and stability time, which was not suitable to end use. The grain test weight, wet gluten content and dough water absorption increased annually, while protein content, dough development time and stability time decreased. The cultivars with good quality had lower kernel weight, which may be the key trait to improve yield. Quality traits had a negative effect on yield and agronomic traits in general, while they can be coordinated to some extent, which had been verified by breeding program. 【Conclusion】Wheat cultivars released recently in Shandong province increased steadily in yield and cultivars with 600 spikes per m2, 40 kernels per spike and 40 mg kernel weight may be more suitable to local weather and production situations. Shandong wheat cultivars released recently had poorer quality in total, and more effective measures should be taken for quality improvement greatly.

Study on the Liquor-Brewing Properties of Waxy Wheat

ZHAO Guo-Jun, XU Zhi-Bin, FENG Bo, WANG Xun, LAN Qiu-Xia, XIANG Chao, HUANG Tian-Fang, WANG Tao

Scientia Agricultura Sinica. 2013, 46(6): 1127-1135. doi:10.3864/j.issn.0578-1752.2013.06.005

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【Objective】The objectives of this study were to reveal the performance of waxy wheat in Chinese liquor brewing process, assess the effects of appearance on liquor quality and explore the ways and methods to apply waxy wheat to liquor-brewing.【Method】Waxy wheat, common wheat and sorghum were used, respectively. The dynamic changes of main chemical compositions and brewing conditions were investigated. The liquor output and liquor quality by using different materials were analyzed.【Result】Waxy wheat has better water absorption capacity, higher absorption rate and higher saccharification temperature than common wheat and sorghum. Under laboratory conditions, the liquor yield and the contents of total acid and total esters in liquor from waxy wheat were superior compared with common wheat and sorghum, while the content of fusel oil was similar. The opposite observation was in the conditions of pilot tests in a winery that fusel oil content and the liquor yield from waxy wheat were higher, while total acid and total esters contents had little differences. In addition, waxy wheat liquor contained minimum aldehydes content. The smell and taste of waxy wheat liquor were the greatest according to tasting notes of professional wine tasters.【Conclusion】It is possible to produce liquor from waxy wheat. By comparing to common wheat and sorghum, waxy wheat liquor has better liquor brewing properties, such as liquor output and better liquor quality. Waxy wheat has a potential for liquor-brewing.

Regional Prediction and Evaluation of Wheat Phenology Based on the Wheat Grow and CERES Models

吕Zun-Fu , LIU Xiao-Jun, TANG Liang, LIU Lei-Lei, CAO Wei-Xing, ZHU Yan

Scientia Agricultura Sinica. 2013, 46(6): 1136-1148. doi:10.3864/j.issn.0578-1752.2013.06.006

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【Objective】The objectives of this study are to investigate the key techniques of wheat phenological model for regional application, and to evaluate the validity of regional application techniques through analyzing the simulated phenology results of WheatGrow and CERES-Wheat at regional scale. 【Method】First, the daily meteorological data at different weather stations were interpolated using the method of Thin Plate Spline (TPS) , and the gridded daily meteorological surface data was generated. Then, the historical sowing dates of wheat at different stations were interpolated using TPS method, and the multi-year average values of interpolated results were generated as regional sowing date surface data. In addition, by integrating the Markov Chain Monte Carlo (MCMC) method with the above two wheat models, the region-specific cultivar parameters were estimated with the measured phenology data from 1998 to 2003, and were taken as the typical cultivar parameters of each province. Finally, the regional daily meteorological data, sowing surface data and region-specific cultivar parameters were input into the above two models, which took grid as basic unit to simulate regional wheat phenology. Meanwhile, the simulated results were evaluated with observed data and the uncertainty coming from cultivar parameters was quantified at site and regional scales. 【Result】The estimated values from the above two models agreed well with the observed values at regional scale. R2 between estimated and observed values of jointing date, heading date and maturity date were 0.85, 0.87 and 0.86 (WheatGrow), 0.87, 0.85 and 0.82 (CERES), respectively. RMSE between the estimated and observed values of jointing date, heading date and maturity date were 9.6, 7.2 and 6.3 d (WheatGrow), 9.4, 7.8 and 6.6 d (CERES), respectively. By analyzing the simulated results of the two models at regional scale, the WheatGrow model had higher prediction accuracy than CERES-Wheat, while the uncertainty came from crop parameters were also higher than CERES-Wheat model.【Conclusion】Through combining the TPS interpolation method with MCMC-based parameter estimation techniques, the mechanism-based model could be effectively extended to regional scales to accurately predict the regional wheat phenology, and quantify the uncertainty coming from the parameter estimation. These results will provide a technology support for estimating crop productivity at regional scale in the future.

Pathways to Synchronously Improving Crop Productivity and Field Water Use Efficiency in the North China Plain

MEI Xu-Rong, KANG Shao-Zhong, YU Qiang, HUANG Yuan-Fang, ZHONG Xiu-Li, GONG Dao-Zhi, HUO Zai-Lin, LIU 恩Ke

Scientia Agricultura Sinica. 2013, 46(6): 1149-1157. doi:10.3864/j.issn.0578-1752.2013.06.007

Abstract ( 929 )

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The North China Plain(NCP), one of the most important agricultural regions in China, is facing a major water-resource crisis. Improving cultivar water use efficiency(WUE), to reduce water use while maintaining high production level, is of great significance for national food and water security. In this review, strategies for crop yield enhancing and water saving in the NCP, and the theoretical and technological problems need to be currently resolved were proposed, referring to the advances in the relevant studies achieved all over the world. The main points include theories and the specific methods for controlling luxurious transpiration of crop population; theories and pathways for reducing field non-productive water consumption; the mechanism and pathways for sustainable utilization of unconventional water that may mitigate regional water crisis; the mechanism and pathways for gaining a balanced improvement of regional water productivity based on the balance between water resources supply and requirement. Finally, the main challenges in basic studies on efficient field water use in the NCP were proposed, and the fields in which important breakthrough may be achieved in the future were also prospected.

Silencing Rice Stripe Virus Gene in Laodelphax striatellus (Fallén) by dsRNA Feeding

CHEN Ai-Ling, LIN Ke-Jian, HE Kang, WANG Gui-Rong, LI Fei

Scientia Agricultura Sinica. 2013, 46(6): 1158-1165. doi:10.3864/j.issn.0578-1752.2013.06.008

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【Objective】 The objective of this study is to use RNA interference (RNAi) technique to explore the feasibility of inhibiting the reproduction of Rice stripe virus (RSV) and to probe the interactions among different RSV genes in vitro. 【Method】 The inhibition function of exogenous dsRNAs to the expression of RSV genes in Laodelphax striatellus (Fallén) were investigated through feeding synthesized dsRNA in vitro. According to the sequences of RSV, three types of dsRNAs (dsRdRp, dsNSvc4, dsCP) were designed and synthesized and then used to feed 2nd instar L. striatellus that infested with RSV. The dsRNA were synthesized by using the T7 RiboMax Express RNAi System. Then 1 µg•µL-1 dsRNA was added to the artificial diet for feeding. The expression level of each RSV gene in insects was examined by quantitative real-time PCR (qRT-PCR) at 96th hour after dsRNA treatment. 【Result】 The results of qRT-PCR indicated that the expression of two RSV genes, NSvc4 and CP, decreased by 93.2% and 94.9%, respectively. Although the decreasing amplitude of RdRp gene was significantly lower, it still decreased by up to 45.6%. Further studies showed that the expressions of other non-target RSV genes decreased when feeding specific dsRNA, which indicated that interaction might exist among different RSV genes. 【Conclusion】 This study offered an important basis for controlling RSV through dsRNA microbe engineering or transgenic rice technology.

Effects of Host-Feeding on Egg Maturation and Longevity of Synovigenic Parasitoid Diglyphus isaea

ZHANG Ying, DANG Guo-Rui, ZHANG Li-Sheng, WANG Meng-Qing, CHEN Hong-Yin

Scientia Agricultura Sinica. 2013, 46(6): 1166-1171. doi:10.3864/j.issn.0578-1752.2013.06.009

Abstract ( 643 )

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【Objective】The objective of this study is to investigate the effect of host-feeding on the egg load and longevity of parasitoid Diglyphus isaea, and to understand the reproductive biology and promote the reproduction potencies of this parasitoid. 【Method】 Female parasitoids were dissected to assess egg load, three factors affecting the egg loads of D. isaea females were examined. These factors are the age of adult parasitoid (i), the host stage utilized (ii) and the size of host-feeding meal (iii). Daily observations were provided the longevity of the host-feeding adult. 【Result】 This parasitoid emerged from their hosts without mature eggs, after given only a carbohydrate diet, they began mature eggs. At day 5, the egg load was 7.5, much higher compared to others, then gradually resorbed after 6 days. The eggs load and eggs laid were significantly higher in parasitoids exposed to 3rd instar larvae of Liriomyza sativae, which were 5.82 and 2.85, respectively. This parasitoid matured more eggs when they were fed on 3rd instar larvae of host on day 3, day 5 and day 7 after emergence. The longevity of the parasitoid feeding on 1st larvae, 3rd larvae and honey feeding was 28.3, 29.26 and 26.8 d, respectively, and no significant difference between them. 【Conclusion】 Host feeding would increase the egg maturation, not prolong the longevity of the parasitoid D. isaea.

Effects of No-Tillage on Supply Characteristics of Soil Inorganic Nitrogen and Rice Yield

WU Ji, GUO Xi-Sheng, ZHANG Xiang-Ming, WANG Yun-Qing

Scientia Agricultura Sinica. 2013, 46(6): 1172-1181. doi:10.3864/j.issn.0578-1752.2013.06.010

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【Objective】To study the effect of no-tillage on supply characteristics of soil inorganic nitrogen and rice yield in paddy-upland rotation system.【Method】A long-term (2007-2010) experiment was conducted the field trail was designed to have two treatments with four replicates: No-tillage; conventional tillage, and the rice cultivar Wandao 68 was used as material.【Result】The results showed that NH4+-N content in arable layer soil was higher at rice early growth stage and lower at the middle and late growth stages than conventional tillage. Compared to conventional tillage, the NO3--N content in 0-10 cm soil layer was increased significantly while NO3--N content in 10-20 cm soil layer of main growth stage was decreased with no-tillage. That could help reduce the leaching loss of NO3--N in soil. The inorganic nitrogen of no-tillage was lower than that of conventional tillage in whole arable layer during rice season. No-tillage enriched soil organic matter and total nitrogen in the surface soil layer. Both the soil organic matter and total N content of no-tillage were significantly higher in 0-10 cm layer and lower in 10-20 cm layer than that of conventional tillage. The soil bulk density in 0-20 cm layer was increased by 3.06%-6.25% compared with conventional tillage. The yield results indicated that no-tillage treatment decreased rice yield notably by 4.11-7.70%. 【Conclusion】In comparison to conventional tillage, no-tillage treatment decreased NH4＋-N content since elongating and resulted in reducing absorption of nitrogen in plants at elongating and heading and the ratio of productive tillers to total tillers significantly. This was the main reason for the reduction of rice yield with no-tillage.

Soil Nitrate N Accumulation Under Different N-Fertilizer Rates in Summer Maize and Its Residual Effects on Subsequent Winter Wheat

ZHANG Jing-Ting, WANG Zhi-Min, ZHOU Shun-Li

Scientia Agricultura Sinica. 2013, 46(6): 1182-1190. doi:10.3864/j.issn.0578-1752.2013.06.011

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【Objective】 The study was conducted to clarify the annual characteristics of soil NO3--N accumulation under different N-fertilizer (NF) rates applied in maize (Zea mays L.) and its residual effects on succeeding wheat (Triticum aestivum L.) in winter wheat-summer maize rotation system.【Method】Four NF rates (0, 120, 240 and 360 kg N•hm-2) were applied in maize and the cultivation practices of water-saving and NF-saving were adopted in wheat, the annual changes of NO3--N in a 2 m-depth soil profile in different NF treatments and its residual effects on wheat were observed.【Result】There was a significant positive correlation between NF rates and residual soil NO3--N amount at maize harvest stage, and the yield of following wheat also showed a significant positive relationship with the residual soil NO3--N amount, which could be fitted by a linear model. Compared with that at maize harvest stage, NO3--N content in each soil layer sharply decreased at wheat maturity stage under the NF rates of 157.5 and zero kg N•hm-2 applied in wheat. 240 kg N•hm-2 applied in maize and 157.5 kg N•hm-2 applied in wheat (N240+157.5) or the treatment N360+0 (360 kg N•hm-2 was applied in maize but no NF in wheat) could meet wheat demand for N, eventually maintained yield, plant N uptake amount and annual apparent N recovery efficiency at a higher level. However, the treatments in which higher NF applied in maize had a great NO3--N leaching loss risk in rainy season. 【Conclusion】 NF applied in summer maize has a strong residual effectiveness to subsequent winter wheat. Adopting the water-saving and NF-saving cultivation techniques in winter wheat not only shrinks sharply the NO3--N accumulation in the soil profile, but also sustains high-yielding and increases NF recovery efficiency. Therefor, the residual effect should be considered in NF optimization in crop production.

Effects of Different Temperature and Light Treatments on Photosynthetic System II in Vitis vinifera L. cv. Cabernet Sauvignon

SUN Yong-Jiang, FU Yan-Dong, DU Yuan-Peng, DI Heng

Scientia Agricultura Sinica. 2013, 46(6): 1191-1200. doi:10.3864/j.issn.0578-1752.2013.06.012

Abstract ( 809 )

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【Objective】The combined effects of different temperature and light treatments on the operation of photosynthetic apparatus were investigated to reveal the mechanism of inhibition of photosynthesis.【Method】One-year-old Cabernet Sauvignon grapevines were treated with different temperatures（28, 34, 37 and 40℃）combined with two kinds of light intensities (800, 1600 μmol•m-2•s-1). Gas exchange and chlorophyll a fluorescence parameters were simultaneously analyzed to investigate the effects of different treatments on the PSII function【Result】Despite the treated light intensities, as temperature increased, net photosynthetic rate (Pn), stomatal conductance (Gs), actual PSⅡ efficiency (ΦPSII), electron transport rate (ETR), performance index (PIABS), density of RCs (RC/CS) and the probability that a trapped exciton moves an electron into the electron transport chain beyond QA (ΨEo) decreased; PSⅡ excitation pressure (1-qP), non-photochemical quenching (NPQ) and approximated initial slope of the fluorescence transient (Mo) increased. Under the moderate light intensities, the decrease of Pn was due mainly to stomatal limitation during the temperature from 28℃ to 37℃, and the xanthophyll cycle played an increased role to protect photosystem II. Under strong light stress, 28℃ treatment decreased Pn to the level of the 40℃ treatment of moderate light intensities, meanwhile PIABS reached a lower level.【Conclusion】Under moderate light intensities, the PSⅡacceptor sides were not injured until the temperature reached 37℃; strong light stress, however, began to injure the PSⅡ acceptor sides from 28℃, when the temperature reached or exceeded 37℃, the density of RCs and the acceptor sides were damaged severely which showed PSⅡcould be more susceptible to strong light than moderate heat temperature.

Respiratory Changes During Dormancy of Grape Buds

CONG Shen, WANG Hai-Bo, WANG Xiao-Di, WANG Bao-Liang, ZHENG Xiao-Cui, SHI Xiang-Bin, LIU Wan-Chun, LIU Feng-Zhi

Scientia Agricultura Sinica. 2013, 46(6): 1201-1207. doi:10.3864/j.issn.0578-1752.2013.06.013

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【Objective】The relationship between respiration and dormancy release of grape bud was discussed for the purpose of laying a theoretical foundation for the artificial control technology of dormancy. 【Method】Respiration inhibitors and oxygen electrode were used to investigate the respiratory changes during dormancy in grape buds (high chilling requirement grape V. vinifera-V.labrusca cv. Summer Black Seedless and low chilling requirement grape V. vinifera cv. Jingmi). 【Result】Different chilling requirement grape cultivars had the similar respiration rate, pathway rate and change tendency. Respiration rate and pathway of low chilling requirement grape Jingmi changed notably about 20 days before high chilling requirement grape Summer Black Seedless, and rapider. The change range of Jingmi was lower than Summer Black Seedless. The variation in total respiratory rate showed single peak curves. EMP (embden-meyerhof-parnas pathway)-TCA (tricarboxylic acid cycle) increased during dormancy release. On level of electron transport, alternative pathway and residual pathway were activated during dormancy release.【Conclusion】Different chilling requirement grape cultivars had different sensitivities, low chilling requirement grape cultivars were more sensitive to low temperature. EMP-TCA was the key of dormancy release. Alternative pathway and residual pathway might be an important role in dormancy release.

Evaluation on Resistance of Grape Gray Mold Pathogen Botrytis cinerea to Pyrimethanil in China

ZHANG Wei, QIAO Guang-Xing, HUANG Jin-Bao, WANG Zhong-Yue, LI Xing-Hong

Scientia Agricultura Sinica. 2013, 46(6): 1208-1212. doi:10.3864/j.issn.0578-1752.2013.06.014

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【Objective】The resistance to pyrimethanil of Botrytis cinerea isolated from diseased grape berries in China was studied.【Method】A total of 104 single conidial isolates of B. cinerea were isolated from diseased grape berries which were collected from vineyards located in 14 different cities or suburban counties in China. The method of mycelial growth inhibition was used to determine the resistance of B. cinerea to anilinopyrimidine fungicides pyrimethanil.【Result】The findings in the current work demonstrated that the resistance frequency of B. cinerea to pyrimethanil was 22.22% to 62.5%. Most of the resistant isolates showed middle resistant or high resistant phenotype, and the resistant frequency of high resistant isolates reached up to 44.23%. Resistance to pyrimethanil of B. cinerea differs in different viticulture climate zones. 【Conclusion】Resistance to pyrimethanil of B. cinerea isolated from grape diseased samples in China is commonly present, and the resistant isolates showed cross resistance. Accordingly, the frequency of use of pyrimethanil should be limited in the prevention of grape gray mold, and fungicides such as dicarboximide and carbamates can be used alternatively. It was suggested that novel fungicides or biofungicides should be introduced for the control of grape gray mold.

Environmental Adaptability of Different Grape Cultivars in Greenhouse

WANG Hai-Bo, WANG Xiao-Di, SHI Xiang-Bin, WANG Bao-Liang, ZHENG Xiao-Cui, LIU Feng-Zhi

Scientia Agricultura Sinica. 2013, 46(6): 1213-1220. doi:10.3864/j.issn.0578-1752.2013.06.015

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【Objective】The objective of this study is to lay a foundation for screening of the grape cultivars which are suitable for the protected cultivation environment.【Method】Twenty-two 5-year-old protected grape cultivars were used as experimental materials to study the adaptability of different grape cultivars under the protected environment and to make clear the important environmental factors which affect high yield in successive years of the protected grapes.【Result】The results showed that the grape cultivars with high-yielding capacity in successive years in the protected cultivation environment conditions have a better ability of resistance to low-light, but not necessarily strong resistance to low concentrations of CO2 and high temperature. Low light is the major environmental factor which affects high yield in successive years of the protected grapes, but low concentration of CO2 and high temperatures are not the major environmental factors.【Conclusion】Hongqitezaomeigui, Zizhenxiang, Centenial Seedless, Wuhezaohong, Hongbiaowuhe, 87-1, Cardinal, Pearl of Csaba, Otilia Seedless, Xiangfei and Hongxiangfei have a higher environmental adaptability and high ability of sustainable productivity. Low light is an important environmental factor which affects high yield in successive years of the protected grapes.

Isolation of Dominant Actinomycetes from Cherry Orchard Soil and Their Functions in Bio-Control and Plant Promotion

LIU Ling-Zhi, QIN Si-Jun, LU De-Guo , WANG Bing-Ying

Scientia Agricultura Sinica. 2013, 46(6): 1221-1229. doi:10.3864/j.issn.0578-1752.2013.06.016

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【Objective】 The dominant actinomycetes in cherry orchard soil were isolated and collected at different growth phases to study their application potential in biocontrol and plant growth promotion. 【Method】 All isolates were assayed for antagonistic activity, fermentation activity in vitro experiments to analyze their antagonistic and growth promoting function. Strain F4-5 with good microorganism inhibiting effect and plant growth promoting functions was explored through 16S rDNA sequence analysis to ascertain its taxonomic status. 【Result】 There were 32 dominant actinomycetes were isolated from Cerasus avium (L.)Moench / C. sachalinensis (L.) Kom. Rhizosphere soil at different growth stages. The number of soil actinomycetes in August was the highest compared with that in May and November. The isolates from different growth stages also varied in their microorganism inhibiting and plant growth promoting capabilities. In general, isolates from August 2010 expressed obvious effect on antagonistic pathogen and plant growth promotion. Among all the 32 isolates, strain F4-5 showed a significant inhibitive effect on 4 tested bacteria including Staphylococcus aureus, Escherichia coli, Bacillus subtilis and B. cereus, as well as 3 tested fungi including Trichoderma viride, Rhizopus stolo and Aspergillus niger, exhibiting a broad-spectrum antibiosis. In the experiment of seeding growth of Malus baccata (L.) Brokh., strain F4-5 also displayed a positive effect on seed germination and root growth, increased by 201.1% and 43.5%, respectively, compared with the control, indicating both antibiosis and plant promoting effects. In addition, strain F4-5 was identified as Streptomyces flavogriseum, according to the morphological characteristics, cultural characteristics and 16S rDNA sequence analysis. 【Conclusion】The actinomycete number, distribution, antibiotic and seed germination promotion in Cerasus avium (L.) Moench / C. sachalinensis (L.) Kom. Rhizosphere soil varied greatly at different stages. The proper strain F4-5 (S. flavogriseum) screened from 32 actinomycete strains showed a good probiotic role and seed germination promotion.

Effect of Packaging on the Postharvest Quality and the Antioxidant Activity of Blueberry

CHEN Hang-Jun, WANG Cui-Hong, GAO Hai-Yan, MAO Jin-Lin, ZHOU Yong-Jun

Scientia Agricultura Sinica. 2013, 46(6): 1230-1236. doi:10.3864/j.issn.0578-1752.2013.06.017

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【Objective】This research was conducted to study the effect of packaging on the postharvest quality and the antioxidant activity of blueberry, and to explore the optimal packaging methods at low temperature. Further theoretical guide for the development and application of storage technique of blueberry.【Method】‘Canlan’ blueberry fruits were stored at (5±0.5)℃ using unpacked, polyethylene (PE) film packaging and perforated PE film packaging. Based on sensory evaluation and analyses of antioxidant enzyme activities and antioxidant contents, the effects of different packaging methods on the postharvest quality and the antioxidant capacity of blueberry were studied. 【Result】The results showed that, as compared to other two packagings, the PE film packaging was significantly better in preventing dehydration and atrophy, slowing down the changes of hardness and the total-soluble solids (TSS), and maintaining the antioxidant capacity of blueberry during storage. However, the PE film packaging showed no significant effect on titratable acid (TA) content. 【Conclusion】 Different packagings exhibited a great effect on the storage quality of blueberry fruit. By the end of the storage period (42 days), the fresh-keeping effect of the PE film packaging method was better than that of the perforated PE film packaging or non-PE-packed method.

Effect of FSH on the mRNA Expression of Cyclin D1 and Cyclin E1 in the Cultured Immature Boar Sertoli Cell

ZHANG Jiao-Jiao, WANG Yi, ZHANG Hui-Qiong, SUN Si, SUN Yan, WANG Xian-Zhong, ZHANG Jia-Hua

Scientia Agricultura Sinica. 2013, 46(6): 1237-1246. doi:10.3864/j.issn.0578-1752.2013.06.018

Abstract ( 634 )

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【Objective】 The objective of the study is to identify whether or how FSH regulates the mRNA expression of cyclin D1 and cyclin E1 in the cultured immature boar Sertoli cell. 【Method】 In the study, the cultured immature boar Sertoli cells were used as materials, and real-time PCR was used to detect the mRNA expression of cyclin D1 and cyclin E1. 【Result】 Different doses of FSH (0-100 ng•mL-1) increased the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), which were peaking at 50 ng•mL-1 (P＜0.05). In addition, FSH (50 ng•mL-1) increased the mRNA expression of cyclin D1 and cyclin E1 in a time-dependent manner (P＜0.05) and peaked at 30 min (P＜0.05). Different doses of Forskolin (0-20 μmol•L-1) increased the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), but lower compared with FSH alone. Rp-cAMP (0-40 μmol•L-1), H-89 (0-30 μmol•L-1) and Verapamil (0-100 μg•mL-1) inhibited FSH induced the mRNA expression of cyclin D1 and cyclin E1 in a dose-dependent manner (P＜0.05), and the combination of Rp-cAMP and Verapamil had more inhibition on FSH activity than individual effect (P＜0.05), but lower than addition of Rp-cAMP and Verapamil. Furthermore, different doses of U0126 (0-10 μmol•L-1) reduced FSH induced the mRNA expression of cyclin D1 and cyclin E1 (P＜0.05), while Rp-cAMP, H-89, Verapamil or U0126 alone had no effect on the mRNA expression of cyclin D1 and cyclin E1 as compared to the control (P＞0.05). 【Conclusion】 FSH regulates the mRNA expression of cyclin D1 and cyclin E1 in a dose- and time-dependent manner, while cAMP-PKA, Ca2+ and extracellular signal-regulated kinase (ERK1/2) is involved in the regulation of FSH on the mRNA expression of cyclin D1 and cyclin E1.

Tissue Expression Profile and Bioinformatics Analysis of Abundance miR-101 from the Hypothalamus of Chicken

LI Guo-Xi, WANG Le-Le, SUN Gui-Rong, KANG Xiang-Tao

Scientia Agricultura Sinica. 2013, 46(6): 1247-1255. doi:10.3864/j.issn.0578-1752.2013.06.019

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【Objective】 The objective of this study is to comprehensively understand the chicken hypothalamus abundance miR-101 expression and potential biological function which will lay a foundation for revealing the molecular mechanisms of hypothalamic regulation of the chicken energy balance.【Method】 Stem-loop quantitative RT-PCR was used to detect the miR-101 expression in 13 different tissues of Gu-shi chicken at four developmental stages. The algorithms PicTar was used to predict miR-101 putative target genes. Gene Ontology, pathway and stratified enrichment were analyzed respectively to target genes. 【Result】 MiR-101 expression exhibitted striking temporal and tissue specificities. The miR-101 expression levels in different tissues of embryonic stages were significantly higher than the expression levels in homologous tissues of hatched chicks, the expression levels in the brain tissues were also significantly higher than the other tissues examined. MiR-101 had abundance enrichment in hypothalamus at 14 d embryonic age and in small intestine and crura muscle at 17 d embryonic age. Predicted target genes of miR-101 were significantly enriched in biological regulation, metabolic processes, developmental processes and cellular processes. Bioinformatics analysis of the nervous system development displayed that the regulatory function of miR-101 was mainly involved in brain development, neuron differentiation, regulation of neurogenesis, glial cell differentiation, astrocyte differentiation, and so on. 【Conclusion】 MiR-101 was the high expression miRNA in brain tissue, and may play an important role in regulating biological processes related to brain development.

Preparation and Characterization of Ractopamine-Imprinted Material Using Surface-Molecular Imprinting Method and Its Adsorption Characteristics

YANG Ting, WU Yin-Liang, LI Cun, ZHAO Jian, CHEN Guo, LU Yan

Scientia Agricultura Sinica. 2013, 46(6): 1256-1262. doi:10.3864/j.issn.0578-1752.2013.06.020

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【Objective】 The objective of this experiment is to prepare a molecule-imprinted material on surface of silica gel for ractopamine. 【Method】 Polymethylacrylic acid (PMAA) was grafted onto the surface of silica gel particles via the coupling effect of γ-methacryloxypropyl trimethoxy silane, and the grafting particle PMAA/SiO2 was prepared. Then, the molecular imprinting towards the grafted PMAA was performed with ractopamine as template molecule and ethylene glycol diglycidylether (EGDE) as crosslinking agent by using the surface-molecular imprinting technique established, and the ractopamine molecule- imprinted material MIP-PMAA/SiO2 was prepared. The morphology of MIP-PMAA/SiO2 was observed by SEM，and the chemical structure of MIP-PMAA/SiO2 was characterized by FT-IR. Static method was adopted to study the binding properties and molecule recognition characters of MIP-PMAA/SiO2 for ractopamine.【Result】The MIP-PMAA/SiO2 on the adsorption ability of ractopamine was significantly better than clenbuterol and salbutamol. The adsorption capacity of the MIP-PMAA/SiO2 was 11.08 mg•g-1, the specificity binding capacity was 9.93 mg•g-1, and the imprinted factor of the polymer was 9.63. The adsorption reaction reached equilibrium after 10 min, and the desorption ration of ractopamine reached 92.3% in 5 min.【Conclusion】The experimental results show that MIP-PMAA/SiO2 have specific recognition selectivity, excellent binding affinity and elution property for ractopamine.

Upregulation and Expression of Bombyx mori bmo-miR-14 and Prediction of Its Target Genes

LIU Yue, YANG Lan-Cui, NIE Zuo-Ming, LU Xuan, LU Zheng-Bing , CHEN Jian, YU Wei, WU Xiang-Fu, ZHANG Yao-Zhou

Scientia Agricultura Sinica. 2013, 46(6): 1263-1271. doi:10.3864/j.issn.0578-1752.2013.06.021

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【Objective】The objective of this study is to offer a method for upregulating miRNA and predicting its targets in BmN cells, and to provide a methodological reference for studying the function of miRNAs in BmN cells. 【Method】An eukaryotic expression vector pSK-hr3-ie1-EGFP-pri-mir-14 for expressing Bombyx mori bmo-miR-14 was constructed and the miRNA gene was expressed in BmN cells. Additionally, the level of bmo-miR-14 in BmN cells was also up-regulated by transfecting chemically synthesized miRNA mimics. Firstly, the fragment containing pri-mir-14 was amplified by PCR and inserted into the downstream of EGFP of eukaryotic expression vector pSK-hr3-ie1-EGFP. The recombinant plasmid pSK-hr3-ie1-EGFP-pri-mir-14, control plasmid pSK-hr3-ie1-EGFP, bmo-miR-14 mimics and negative control mimics were transfected into BmN cells, respectively. The fluorescent light was observed and used to identify the efficiency of transfection. The miRNA levels in transfected BmN cells were identified by qRT-PCR. The RNAhybrid and miRanda were used to predict the targets of bmo-miR-14, respectively. 【Result】The miR-14 level was improved in BmN cells by transfecting with pSK-hr3-ie1-EGFP-pri-mir-14 and bmo-miR-14 mimics, and the level was improved as 2.1 and 984 times as the control, respectively. Further, the targeted genes of bmo-miR-14 were identified by bioinformatics tools and a total of 153 and 171 potential targeted genes were identified by RNAhybrid and miRanda software, respectively. In the predicted target genes, a total of 49 genes were predicted by RNAhybrid and miRanda together. The 49 targeted genes were assigned to GO terms of molecular function ontology, and binding and catalytic were overrepresented. In the biological process ontology, cellular process and metabolic process overrepresented among the 49 genes.【Conclusion】The approach to upregulate the level of bmo-miR-14 by transfecting with recombinant expression vector or miRNA mimics has laid a good foundation for the further studies on bmo-miR-14 in BmN cells.

Study on Induction of Porcine Putative Embryonic Germ Cells into Nerve Cells in vitro

CONG Yi-Mei, MA Jing, SUN Rui-Zhen, WANG Jian-Yu, XUE Bing-Hua, HU Kui, YIN Zhi, LIU Zhong-Hua

Scientia Agricultura Sinica. 2013, 46(6): 1272-1279. doi:10.3864/j.issn.0578-1752.2013.06.022

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【Objective】The objective of the study is to detect the potential of porcine putative embryonic germ cells differentiating into nerve cells. 【Method】Co-culture of Gonad-mesonephros (GM) region stromal cells and primordial germ cells was used to get porcine putative embryonic germ cells. Directional differentiation method was adopted for neural differentiation. 【Result】 Compared with MEF , GM region stromal cells as feeder could support the growth of porcine putative embryonic germ cells with no significant difference in the proliferation of embryonic germ cells. Porcine putative embryonic germ cells had strong AP activity, Q-PCR results indicated the expression of Oct4, Sox2 and Nanog genes. Proliferating ability of porcine putative embryonic germ cells showed a "S" type, namely the growth of latency, logarithmic phase and plateau phase. Embryoid body could be derived after suspending culture of porcine putative embryonic germ cells in vitro. After induction, porcine putative embryonic germ cells could differentiate into a variety of neural cell types with expression of neural stem cells, neurons and glial cells markers.【Conclusion】These results indicate that porcine putative embryonic germ cells can be derived from porcine early gonadal and have the ability of differentiating into a variety of neural lineage cells in vitro.

The Research and Control Situation of Ecology and Animal Poisoning of Locoweed in Western Natural Grassland of China

ZHOU Qi-Wu, ZHAO Bao-Yu, LU Hao, WANG Shan-Shan, ZHANG Liang, WEN Wei-Li, YANG Xiao-Wen

Scientia Agricultura Sinica. 2013, 46(6): 1280-1296. doi:10.3864/j.issn.0578-1752.2013.06.023

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Natural grassland is an important production base of grassland and animal husbandry, and it is also a very important ecological barrier in China, so it is a great strategic importance for China to maintain ecological and food security and protect survival environment of human beings and fulfill permanent utilization of grassland. For a long time, some natural and human factors lead to grassland desertification and contagious poisonous weeds in the rangeland, such as grassland drought, overgrazing, blind reclamation, shortages of fund and lagging management. Especially in the last few decades, the rapid spreading of poisonous-weeds have caused some ecological problems such as decrease of vegetation community and biodiversity, single vegetation and desertification, and it seriously affects the grassland ecological balance and the sustainable development of local animal husbandry. “Locoweeds” has been a primary poisonous weed in the western natural grassland, and it brings about a billion RMB in economical losses every year. The research history, species and ecological distribution and the situation of the toxicity disaster, the toxic ingredients and mechanism of locoweeds in grassland are summarized in this paper, and the problems of locoweed research in the future are given, it is of great significance for understanding the ecological function of locoweeds and comprehensive control of the locoweed disaster in western natural grassland of China.

Effects of Different LED Light Qualities on Cucumber Seedling Growth and Chlorophyll Fluorescence Parameters

CAO Gang, ZHANG Guo-Bin, YU Ji-Hua, MA Yan-Xia

Scientia Agricultura Sinica. 2013, 46(6): 1297-1304. doi:10.3864/j.issn.0578-1752.2013.06.024

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【Objective】 The differences of cucumber seedling growth, as well as variances of chlorophyll content and chlorophyll fluorescence parameters were studied. 【Method】Red light, blue light, red+blue (R+B) light generated by light-emitting-diode, white light as control, were applied to study the influences of different LED light qualities such as W, R, B, 7R/3B, 8R/2B, 9R/1B on cucumber seedling leaves with Yinpei 99 as materials. 【Result】 The differences of cucumber seedling showed that although treatments by single red light, blue light or white light had no significant effect on plant height and leaf length or width, obvious increase was observed about stem diameter, contents of chlorophyll and carotenoid in the blue light treatment. Moreover, the highest biomass, and NPQ were observed in red light treatment, with typical character of sun plants due to higher ratio of chlorophyll a/b. Furthermore, it was showed that a several combination of red+blue (R+B) treatment was beneficial to the growth of cucumber seeding, and seedling index, dry plant weight, Fv/Fm、ФPSII and qP were significantly higher than those of single red or blue light treatment. 【Conclusion】 It was concluded that 8R/2B treatment showed its superiority with significant increase of plant height, seedling index, dry plant weight as well as Fv/Fm, ФPSII and qP, so 8R/2B treatment enhanced the opening degree, the light energy conversion efficiency of photosystemⅡ, and protected the effect of photosystemⅡ.

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