Scientia Agricultura Sinica

Conditional and Unconditional QTL Mapping of Grain Starch Accumulation in Wheat

TIAN Bin, LIU Bin, ZHU Zhan-Ling, XIE Quan-Gang, TIAN Ji-Chun

Scientia Agricultura Sinica. 2011, 44(22): 4551-4559. doi:10.3864/j.issn.0578-1752.2011.22.001

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【Objective】Conditional QTL is a method of studying the dynamic expression of quantitative traits genes, and it was applied to better reveal dynamic expression of gene in wheat starch.【Method】QTLs for developmental behavior of wheat grain starch content (GSC) were mapped using a double haploid (DH) population with 168 progeny lines, derived from a cross between two elite Chinese wheat cultivars Huapei 3×Yumai 57. GSC were evaluated at five seed filling stages (12d, 17d, 22d, 27d, and 32d after ?owering) in six different environments. This study could attain more information to understand the dynamic effects of corresponding QTLs at different developmental stages as well as the genetic information of GSC accumulation.【Result】The results showed that seven unconditional QTLs and four conditional QTLs were detected in different seed filling periods totally, but no one conditional QTL was detected in all periods. Seven unconditional QTLs were detected on chromosomes 2A, 3A, 3B, 4A and 5D, the unconditional QTL named QGsc4A persistently expressed at all seed filling stages, while unconditional QTLs of different seed filling periods could explain 13.57%, 16.57%, 21.96%, 22.53%, 22.90% of phenotypic variance respectively. Among the four conditional QTLs, QGsc4A was detected 12d, 17d, and 32d after ?owering, which accounted for 21.80% of phenotypic variation and played the most important role for the accumulation of GSC. The rest unconditional QTLs and conditional QTLs were only detected in a special or few periods, and there was no conditional QTL detected up to 27 days after ?owering. 【Conclusion】QTL analyses were performed using a mixed linear model approach, which suggest that the QTLs express depending on different developmental stages. The present data provide new insights into understanding the genetic mechanism and regulation network underlying the development of GSC in wheat, which is beneficial to genetic manipulations for yield.

QTL Analysis of Two-Seed Pod Storage Capacity in Soybean in Different Years

YANG Zhen, SHEN Yan-Ru, HAN Dong-Wei, LIU Chun-Yan, JIANG Hong-Wei, CHEN Qing-Shan, HU Guo-Hua

Scientia Agricultura Sinica. 2011, 44(22): 4560-4569. doi:10.3864/j.issn.0578-1752.2011.22.002

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【Objective】The objective of this study is to locate consensus QTLs of two-seed pod length and width for breeding new varieties with high two-seed pod storage capacity and increase yield of soybean. 【Method】In order to find out the steady and repeatable QTLs of these traits, a F2:14-F2:18 RIL population containing 147 lines derived from a cross between Charleston as female and Dongnong594 as male parent were used in this experiment. A genetic linkage map was constructed with 164 SSR primers screened in two parents and amplified in 147 lines population. Two-seed pod length and width showing polymorphic in parents were investigated and QTLs were analyzed in one site in the early two years and two sites in the late three years. 【Result】 Nineteen QTLs for the two-seed pod length and width were analyzed with software Windows QTL Cartographer V2.0 by CIM and MIM under different environments, pod length were mapped in the linkage groups A1, B2, C2, D1a, D1b, N and G, 17 QTLs for two-seed pod width were mapped in the linkage groups A1, C2, D1a, D1b, N and H. In these QTLs,7 QTLs for two-seed pod length with linkage primers including Satt200—qTSPL-a1-1—Satt042, Sat_214—qTSPL-d1a-1—Sat_112, Satt198—qTSPL-d1a-3—Satt502, Satt370— qTSPL-d1a-6—Satt402, Sat_092—qTSPL-c2-4—Satt289, Satt277—qTSPL-c2-5—Sct_188 and Satt168—qTSPL-b2-1—Sat_083 could be detected together by CIM and MIM, 1 QTL for two-seed pod width including Satt528—qTSPW-d1a-2—Satt182 could be detected together by two methods. Eight two-seed pod length QTLs including Satt200—qTSPL-a1-1—Satt042, Sat_119— qTSPL-a1-2—Sat_105, Sat_214—qTSPL-d1a-1—Sat_112, Satt220—qTSPL-d1a-4—Sat_162, Satt370—qTSPL-d1a-6—Satt402, Satt168—qTSPL-b2-1—Sat_083, Sat_092—qTSPL-c2-4—Satt289 and Satt277—qTSPL-c2-5—Sct_188 could be detected in more than two years, 4 two-seed pod width QTLs including Satt076—qTSPW-c2-1—Satt072, Satt335—qTSPW-c2-2—Sat_120, Satt200— qTSPW-a1-1—Satt042 and Satt182—qTSPW-d1a-3—Satt584 could be detected in more than two years.【Conclusion】Mapping two-seed pod length and width QTL provide valuable information for soybean molecular marker assistant breeding selection.

Differences of Nitrogen Uptake and Utilization of Conventional Rice Varieties with Different Growth Duration

DONG Gui-Chun, WANG Yi, YU Xiao-Feng, ZHOU Juan, PENG Bin, LI Jin-Qian, TIAN Hao, ZHANG Yan, YUAN Qiu-Mei, WANG Yu-Long

Scientia Agricultura Sinica. 2011, 44(22): 4570-4582. doi:10.3864/j.issn.0578-1752.2011.22.003

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【Objective】 The objective of the experiments is to investigate the differences in nitrogen uptake and utilization and analyze the ways to improve nitrogen uptake and utilization of the conventional rice varieties with different growth duration (GD). 【Method】 A total of 88 and 122 conventional indica rice varieties were used in 2001 and 2002 and 94 conventional japonica rice varieties were used in 2008 and 2009 in solution culture experiments. The GD, dry matter weight of each plant organ (including root), nitrogen content and grain yield and yield components were measured. Based on GD (from sowing to heading stage) by the MinSSw (dynamic clustering method-minimum sum of squares within groups) method, these rice varieties were clustered into 6 types in the alignment from low to high (A, B, C, D, E and F) for determining the differences in nitrogen uptake and utilization and analyzing their causes. 【Result】The rice varieties with longer GD had larger amount of nitrogen absorption (ANU) (indica rice varieties) or comparatively larger ANU (japonica rice varieties) at heading stage and maturing stage, however it is not predominant of ANU at grain filling stage. The rice varieties with longer GD showed a lower nitrogen concentration (NC) (japonica rice varieties) or no difference in the NC among varieties (indica rice varieties), also had less panicle numbers per unit area (indica rice varieties) or less difference in the panicle numbers among varieties (japonica rice varieties). However they had longer growing days, higher dry matter weight, higher ANU per single panicle, stronger nitrogen uptake intensity per single panicle (indica rice varieties) or comparatively higher nitrogen uptake intensity per panicle (japonica rice varieties). The dry matter weight, the ANU per single panicle, nitrogen uptake intensity per single panicle had more impact on the amount of nitrogen uptake than nitrogen concentration, panicle numbers per unit area, growing days in both indica and japonica rice varieties. The rice varieties with longer GD had low nitrogen use efficiency for grain yield (NUEg) (indica rice varieties) or over medium NUEg (japonica rice varieties) and comparatively higher nitrogen use efficiency for dry weight (japonica rice varieties). And the rice varieties with longer GD showed a higher nitrogen distribution percentage in leaf and stem and sheath at heading and maturity stages, and lower or comparatively lower nitrogen ratio in panicle (japonica rice varities at ripening stage). 【Conclusion】 The rice varieties with longer GD had stronger nitrogen uptake ability (indica rice varieties) or comparatively stronger nitrogen uptake ability (japonica rice varieties) at heading and maturing stages, and had lower NUEg (indica rice varieties) or over medium NUEg (japonica rice varieties). Although, the rice varieties with longer GD had low nitrogen concentration or panicle number, they had longer growing days, larger dry matter weight, higher and comparatively higher amount of nitrogen uptake per single panicle and nitrogen uptake intensity per single panicle. Improvement of dry matter weight, nitrogen uptake ability per single panicle and nitrogen uptake intensity per single panicle were helpful to improve the amount of nitrogen uptake. The promotion of nitrogen translocation from vegetative organs to panicle, and the decrease of the nitrogen percentage remained in sheath and stem and leaves were both beneficial to improving nitrogen use efficiency in rice varieties (indica rice varieties or japonica rice varieties) with longer GD. For japonica rice varieties, the low nitrogen concentration of rice plant at maturity was also one of the important factors on improving nitrogen use efficiency.

Variation of Cropland Phenology in Mid-eastern Inner Mongolia

TAO Jian, ZHANG Ge-Li, WANG Jun-Bang, DONG Jin-Wei

Scientia Agricultura Sinica. 2011, 44(22): 4583-4592. doi:10.3864/j.issn.0578-1752.2011.22.004

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【Objective】 Using RS technique, this paper monitored phonological trend of cropland in mid-eastern Inner Mongolia, and analyzed the influences of temperature and precipitation on phenology thereby potential influences on net primary productivity (NPP). 【Method】 Variation of cropland phenology and productivity was analyzed by trend analysis. The influences of climate change on NPP were explored by spatial correlation and significance analysis. 【Result】 Cropland phenology of most cropland lengthened obviously. Regions with advanced starting and delayed ending occupied the maximal percentage, followed by delayed starting and advanced ending, phenology variation was sensitive to temperature and precipitation spatially, growing season was shortened by precipitation significantly thereby resulting in decrease and large fluctuation of NPP. Inter annual NPP of cropland significantly influenced by temperature increased steadily and apparently. 【Conclusion】 Cropland phenology could be addressed by remotely sensing technique. Response of cropland phenology to temperature and precipitation evidently, and NPP was affected by climate change according to that.

Development of a SSR Molecular Marker for Puccinia graminis f. sp. tritici

WANG Xi, LIU Tai-Guo, XIANG Wen-Sheng, CHEN Wan-Quan

Scientia Agricultura Sinica. 2011, 44(22): 4593-4599. doi:10.3864/j.issn.0578-1752.2011.22.005

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【Objective】The objective of this study is to build a simple, quick and accurate molecular technique to detect Puccinia graminis f. sp. tritici and forecast this destructive disease. 【Method】A microsatellite-enriched genomic library of Pgt was constructed by using the method of FIASCO. Based on the sequence of Pgt microsatellite, pairs of specific SSR primers were developed and screened. 【Result】The primer Pgtfssr1 (f/r) generated a polymorphic pattern displaying a 395 bp DNA fragment specific for Pgt, whereas no DNA fragment was obtained in other 24 non-target wheat fungal pathogens. The existence of specific DNA fragment was detected in the infected wheat tissues at 30 h post-inoculation, and the sensitivity of this molecular marker was Pgt DNA template of 1 ng•μL-1.【Conclusion】A specific SSR marker for the detection of wheat stem rust has been successfully developed, which could be used for the early diagnosis and forecast of wheat stem rust.

Effects of Tobacco Plants Infested by Bemisia tabaci (Gennadius) B-biotype on the Development and Reproduction of Spodoptera litura (Fabricius) and Related Mechanisms

WANG Hong-Tao, XUE Ming, LI Qing-Liang, ZHANG Xiao, ZHOU Fang-Yuan

Scientia Agricultura Sinica. 2011, 44(22): 4600-4609. doi:10.3864/j.issn.0578-1752.2011.22.006

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【Objective】 The objective of this study is to define defense response of tobacco plants to Spodoptera litura induced by Bemisia tabaci B biotype and their spatial effect, and to explore its role in interspecific competition between B. tabaci and S. litura and its possible mechanism. 【Method】 The effects of leaves in different positions of tobacco preinfested by B. tabaci B-biotype on growth, development and fecundity of S. litura were investigated by feeding method and the change of related defense enzymes was analyzed using biochemical method in laboratory. 【Result】 The results showed that tobacco plants preinfested by B. tabaci B-biotype feeding had a negative effect on growth, development and fecundity of S. litura, and different leaf positions of treated plants reacted differently, the sequence was the whitefly-infested leaves＞the middle leaves＞the white-vein leaves. There were significant differences between treatment and its control. Trypsin protease inhibitor (TPI) content, β-1,3-glucanase activity and chitinase activity in the whitefly-infested leaves, the middle leaves and the white-vein leaves of the treated tobacco plants all increased significantly. The unexpanded leaves showed no significant effect on both the growth, development and fecundity of S. litura and the change of related defense enzymes. 【Conclusion】 Tobacco preinfested by B. tabaci B-biotype had a negative effect on growth, development and fecundity of S. litura. Furthermore, this effect was systemic and could be transported through the plant. Infestation by B. tabaci B-biotype in tobacco plants led to the higher induction of TPI, β-1, 3-glucanase and chitinase, and they may have a correlation with the negative effect on S. litura. The defense responses of tobacco plants induced by B. tabaci B-biotype feeding were considered to play an important role in the host defense against S. litura.

Response of Abundance and Composition of the Bacterial Community to Long-term Fertilization in Paddy Soils

YUAN Hong-Chao, QIN Hong-Ling, LIU Shou-Long, TONG Cheng-Li, WEI Wen-Xue, WU Jin-Shui

Scientia Agricultura Sinica. 2011, 44(22): 4610-4617. doi:10.3864/j.issn.0578-1752.2011.22.007

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【Objective】 The impacts of long-term fertilization on soil microbial community structure and abundance were studied to clarify their relationship and provide a theoretical basis for the soil fertility evaluation and management of rice cultivation. 【Method】 The soil samples were collected from long-term fertilization experimental plots receiving no fertilization, chemical fertilization, chemical fertilizer + crop residue incorporation. The terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR analysis were combined to characterize the bacterial community structure and abundance.【Result】Proteobacteria (150 bp; relative abundance 33%—37%) and actinomycetes (67 bp; relative abundance 20%—25%) dominated the bacterial community in paddy soils based on T-RFLP analysis. There was a pronounced difference in the bacterial community composition and diversity responding to the long-term fertilization regimes. CCA results indicated that soil organic carbon (SOC) and pH had the greatest influence on the bacterial community composition. The diversity analysis of Shannon and Evenness Index showed that compared with the unfertilized control, all fertilization treatments significantly increased the bacterial diversity and the harvested residue incorporation also shared the highest. The abundance of bacterial 16S rDNA ranging from 4.34×1010—10.94×1010 copies•g-1soil was determined by real-time PCR, and the amount of 16S rDNA copy number in the fertilization incorporations increased about 50% to 100% of which in unfertilized treatments 【Conclusion】Long-term chemical fertilizer and crop residue incorporation improved soil quality and fertility, and consequently resulted in the shift of the bacterial microecology and promoted the bacterial diversity and abundance.

Characteristics of N Absorption, Utilization and Optimum N-fertilizer Rate Based on a Long-Term Fertilization Experiment of Oats

XIAO Xiang-Fen, ZHOU Chuan-Jiao, ZHOU Shun-Li, HU Yue-Gao, REN Chang-Zhong, GUO Lai-Chun, WANG Chun-Long

Scientia Agricultura Sinica. 2011, 44(22): 4618-4626. doi:10.3864/j.issn.0578-1752.2011.22.008

Abstract ( 850 )

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【Objective】The objective of this paper was to clarify the characteristics of N absorption and utilization, and determine the optimum N-fertilizer (NF) rate in oats in Baicheng, Jilin province. 【Method】Husked oat cultivar Baiyan 7 and naked oat cultivar Baiyan 2 were used and five NF rates (0, 30, 60, 90 and 120 kgN•hm-2) were designed, a long-term fertilization experiment in which the NF rate in each plot was kept the same as its original designed amount throughout the research period was conducted in a randomized block design.【Result】N amount (NA) accumulated in different growing periods or in the whole life, NA accumulated in panicle, NA transferred from vegetative organ to grain and NA absorbed after anthesis increased with the increase of NF rate, and there was a significant difference among the NF treatments. The percentage of NA accumulated in seedlings was higher but lower in late grain filling stage in no or lower NF treatment. The rate of N transferred from vegetative organ ascended firstly and then descended with the increase of NF rate. Apparent NF recovery efficiency and agronomic efficiency of applied NF declined with the increase of NF rate. The maximum N harvest index was observed in lower or medium NF treatment, and the minimum in no or the highest NF treatment. There was a significant interannual difference in N physiological use efficiency, and a significant interaction between year and cultivar was also found. N fertilization increased oats grain yield significantly (P＜0.05), but there was no significant difference between the treatments of 90 kgN•hm-2 and 120 kgN•hm-2. The NF rate at which the highest grain yield was harvested increased year by year in the continuous in-situ study. 【Conclusion】The characteristics of N absorption and utilization of oats was significantly affected by N fertilization, and there was a difference between the two cultivars. Baiyan 7 was more sensitive than Baiyan 2 in response to NF rate. A high-yielding and high N use efficiency production of oats could be achieved at the N rate of 90 kg•hm-2 in Baicheng, Jilin province.

Effect of Elevated Atmospheric CO2 and N Fertilization on Soil Nematode Community in a Rice-Wheat Rotation System

BAO Xue-Lian, LI Qi, LIANG Wen-Ju, ZHU Jian-Guo

Scientia Agricultura Sinica. 2011, 44(22): 4627-4635. doi:10.3864/j.issn.0578-1752.2011.22.009

Abstract ( 662 )

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【Objective】 The effect of elevated CO2 and N fertilization on the below-ground ecosystem was studied. 【Method】 A study was conducted in the rice-wheat rotational FACE platform in China to quantify the effect of elevated CO2 and N fertilization on soil nematode community.【Result】Thirty-five genera and 15 functional guilds were observed in this system, and Acrobeloides, Eucephalobus, Filenchus and Hirschmanniella were dominant genera. Filenchus and Eucephalobus were sensitive to the treatment of elevated CO2 and N fertilization, respectively. Under the FACE condition, the abundance of total nematodes, functional guilds of Fu2 and Om4 increased significantly. The interactive treatment of elevated CO2 and N fertilization influenced the abundance of Fu2. 【Conclusion】Elevated CO2 and N fertilization resulted in the changes of soil nematode community structure and decomposition pathways. Under the high N conditions (HN), elevated CO2 decreased the channel index (CI), and bacteria dominated decomposition pathway was found in the FACE (HN) treatments in the wheat season. Different N fertilization managements influenced the values of structural index (SI), and the higher SI was observed under the low N condition (LN), which suggested a stable ecologically successional status with less disturbance.

Effects of Watermelon Cultivars with Different Resistances to Fusarium oxysporum f. sp. niveum and Grafting on Rhizosphere Soil Microorganism Population and Community Structure

WU Feng-Zhi, AN Mei-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4636-4644. doi:10.3864/j.issn.0578-1752.2011.22.010

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【Objective】 The purpose of this study is to reveal the soil microbiology mechanism of grafting on increase of plant disease resistance, and the differences of rhizosphere soil microorganism population and community structure of grafted and ungrafted watermelon cultivars with different resistances to Fusarium oxysporum f.sp. niveum were evaluated. 【Method】 Changes of rhizosphere soil microorganism population and community of own-rooted watermelon cultivars (Tianniu, resistant cultivar; and Tianshi, susceptible cultivar), rootstock (Boqiang No.1, pumpkin) and their grafted plants were investigated by plate culture method and PCR-DGGE method at different watermelon growth stages. 【Result】 Plate culture results showed that the rhizosphere culturable bacteria population of resistant watermelon cultivar was significantly larger than that of the susceptible cultivar, while the fungi population was significantly smaller than that of the susceptible cultivar of the watermelon at middle-late growing stage. The rhizosphere culturable bacteria and actinomyces populations of the grafted susceptible cultivar were larger than the ungrafted one, but the Fusarium population in rhizosphere soil of grafted watermelon was smaller than that of the ungrafted. PCR-DGGE results showed that rhizosphere soil bacteria and fungi community structures of all treatments changed with watermelon growth stages, and were different between different treatments. Further sequencing analysis of specific bands from DGGE profiles indicated that different groups of bacteria and fungi occurred in the rhizosphere of different watermelon cultivars and rootstock Pupkin cultivar. 【Conclusion】 The resistant watermelon cultivar had significant higher quantities of rhizosphere soil bacteria and actinomyces compared with the susceptible one, while the resistant watermelon cultivar had significant lower quantities of rhizosphere soil fungi and Fusarium compared with the susceptible one. The resistance of watermelon cultivar was closely related to the size of rhizosphere microorganism population. The grafting treatment significantly improved rhizosphere soil bacteria and actinomyces quantities and reduced the quantities of rhizosphere soil Fusarium of susceptible cultivar. The grafting treatment also changed soil microbial community structures. It was concluded that plant genotype has a significant impact on soil microbial community structure, and the differences in the rhizosphere microbial community may contribute to the differences in resistance to F. oxysporum f. sp. Niveum and cultivars.

Factors Influencing the Occurrence and Development of Rhizoid and Protocorm-Like Body in Rosa spp.

ZHANG Jian-Fu, BI Ling, CHEN Xiao-Li, GUO Yan-Chao, ZHAO Liang-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4645-4652. doi:10.3864/j.issn.0578-1752.2011.22.011

Abstract ( 919 )

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【Objective】 Leaf explants of multiple species of Rosa spp. were used for investigation of the factors influencing the occurrence and development of rhizoid and protocorm-like body in this paper，which can lay a foundation for revealing the occurrence mechanism of PLB and molecular breeding. 【Method】 The growth of rhizoid and protocorm-like body were observed and statistics were made by changing the medium components, different growth regulators and their concentrations, culture conditions and the integrity of the rhizoid on the Murashige and Skoog (MS) medium.【Result】The results showed that the occurrence and size of the rhizoid was different depending on the type of auxin, but not cytokinin. Rhizoid could be induced from leaf explants of Rosa spp. on the Murashige and Skoog (MS) medium containing any type of the three auxins. On the other hand, rhizoid could be induced from the leaf explants only in dark, however, this process was significantly suppressed under light condition. On the contrary, lighting promoted the matured rhizoid turning green and developed into protocorm-like body(PLB), but this process did not happen under dark condition and light-dark condition without TDZ. Cutting part of the matured rhizoid from the top, the rhizoid could also develop into PLBs, whereas, the rhizoid cut from the callus surface, couldn’t grow into PLBs, but rather turn brown and died, whether they were placed horizontally or vertically. This indicated that the tip of the rhizoid was not essential for the PLBs formation, but the bottom of rhizoid was vitally important for TDZ absorption, loss of the callus on the rhizoid bottom affected the absorption and transportation of TDZ, and further, the development of the protocorm-like body was suppressed. 【Conclusion】 This method is of universality for the rhizoid induced by auxin. Lighting is a inhibitor for the occurrence of the rhizoid but a promoting factor for the information of the PLB, lighting and TDZ has a synergistic action on the process of rhizoid developing into protocorm-like body. The tip of rhizoid is not the essential for the PLBs formation, but the bottom of rhizoid is very important for TDZ absorption.

The Origin Discrimination of Tartary Buckwheat Based on the Mineral Elements

ZHANG Qiang, LI Yan-Qin

Scientia Agricultura Sinica. 2011, 44(22): 4653-4659. doi:10.3864/j.issn.0578-1752.2011.22.012

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【Objective】The characteristics of mineral elements in tartary buckwheat from different provinces were analyzed in order to make choice of the effective index in tartary buckwheat origin discrimination, and to explore the method of tartary buckwheat origin traceability and discrimination.【Method】Seven mineral element contents (Cu, Zn, Fe, Mn, Ca, P and Se) were analyzed in thirty-nine samples from five provinces of China（i.e., Shanxi, Gansu，Qinghai，Sichuan and Yunnan）. On the basis of stepwise selection，the K Nearest Neighbor (KNN) analysis of nonparametric discriminant was applied to data analysis.【Result】The results showed that the mineral element contents were different in tartary buckwheat varieties from different provinces. In conclusion, element contents of Cu and P were the highest in the Tartary buckwheat varieties from Yunnan while Se was the highest in those from Shanxi. However, the contents of Zn，Fe and Ca were the highest in those from Qinghai, while Cu, Zn, Fe, Ca and P were the least in those from Sichuan, and Mn was the least in those from Gansu. Then, mineral element character indexes (Zn, Mn, Ca, P and Se) affecting the classification greatly were selected with discrimination analysis. A percentage of 97.4% of correct classification was achieved by resubstitution and cross-validated.【Conclusion】The determination of mineral element content in combination with modern statistical techniques should be a useful and convenient tool for the origin discrimination and standardization of tartary buckwheat.

Phenolic Profiles and Antioxidant Activity of Momordica charantia L. of Different Varieties

HUANG Long, DENG Yuan-Yuan, ZHANG Ming-Wei, ZHANG Yan, WEI Zhen-Cheng, ZHANG Rui-Fen, TANG Xiao-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4660-4668. doi:10.3864/j.issn.0578-1752.2011.22.013

Abstract ( 1023 )

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【Objective】 The content of free and bound phenolics and the antioxidant activity of the bitter melon flesh of different varieties were compared. 【Method】 The content of free and bound phenolics and the free flavonoids and the composition of the phenolics of free fraction of the bitter melon flesh of fourteen different varieties were determined. Also, the FRAP, DPPH and ABTS+• antioxidant index were adopted to evaluate the antioxidant capacities of free phenolics.【Result】The free, bound and total phenolic content of 14 bitter melon cultivars ranged from 157.58 to 382.92, 6.46 to 54, and 175.27 to 413.79 mg of gallic acid equivalents/100 g dry weight (DW), respectively. The coefficients of variation (CVs) of free, bound and total phenolic content in bitter melon varieties was 23.50%, 61.04%, and 21.58 %, respectively. The percentage contribution of free phenolics to the total was 91.34%, and that of bound phenolics to the total was only 8.66%. The free flavonoid content of bitter melon samples ranged from 8.97 to 18.22 mg CE/100 g DW, The CV of free flavonoid content in bitter melon varieties was 22.80%. Vanillic acid, epicatechin and rutin were detected in bitter melon samples and ranged from 1.83 to 9.29, 35.17 to 114.52, and 0.91 to 4.53 mg/100 g DW, respectively. The CVs of vanillic acid, epicatechin and rutin content in bitter melon varieties were 43.85%, 26.97%, and 33.09%, respectively. The result of FRAP of 14 bitter varieties varied from 272.16 to 713.32 mg trolox equivalents/100 g DW and the CV was 27.67%. The IC50 values of DPPH of 14 bitter samples ranged from 11.43 to 34.14 mg GAE/100 g DW and the CV was 35.10%. The IC50 values of ABTS+• of 14 bitter samples varied from 21.57 to 119.71 mg GAE/100 g DW and the CV was 63.75%. There was a significantly positive correlation between free phenolic content and FRAP values of 14 bitter samples (P＜0.01). While there existed a significantly negative correlation between free phenilic content and the IC50 values of DPPH and ABTS+• of 14 bitter samples, respectively (P＜0.01).【Conclusion】These results indicate that there are significant differences in phenolic profilest and antioxidant activity of bitter melon among different cultivars and phenolics are the mainly active compound of antioxidant activity. Interestingly, the phenolics of bitter melon flesh are mainly present in free form.

An Indirect and Accurate ELISA for Detection of Alicyclobacillus acidoterrestris in Apple Juice Concentrate

LI Jian-Ke, WANG Feng, XIA Kai

Scientia Agricultura Sinica. 2011, 44(22): 4669-4677. doi:10.3864/j.issn.0578-1752.2011.22.014

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【Objective】 An establishment of a rapid and accurate indirect ELISA for detection of Alicyclobacillus acidoterrestris in apple juice concentrate(AJC) was studied. 【Method】 Polyclonal antibody was generated by immunizing rabbits with A. acidoterrestris separated from AJC and indirect enzyme-linked immunosorbent assay was performed to analyze the antigenicity of A. acidoterrestris and its spores. 【Result】 The new indirect ELISA was used successfully in detecting A. acidoterrestris for the first time, and the detection limit was 5×104 CFU/mL. In the detection of AJC, the samples were filter sterilized and enriched, followed by the indirect ELISA assay. This method could be performed in less than 24 h, and the minimum detection limit has reached the international standard for detecting A. acidoterrestris, which AJC is less than 1 CFU/10 mL. The result gained by indirect ELISA was in accord with that by KFL, a conventional culture assay. 【Conclusion】 The developed indirect ELISA has good specificity, sensitivity and reproducibility for the detection of A. acidoterrestris in juice concentrate, which is accurate, convenient, and quick.

Effect of Nitrogen Levels of Diets on Ruminal Fermentation Characters and Nitrogen Fractions in Yak

MI Jian-Dui, GUO Xu-Sheng, ZHOU Jian-Wei, XIN Guo-Sheng, ZHANG Ying, ZHU Yu-Huan, LONG Rui-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4678-4686. doi:10.3864/j.issn.0578-1752.2011.22.015

Abstract ( 638 )

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【Objective】 The objective of this study is to evaluate the effect of different N levels of diets on the ruminal fermentation and nitrogen fractions in yak. 【Method】 Four castrated yaks at (148.5±9.2) kg and 3-year-old of age were used in a partially replicated 4×4 Latin square with 4 different N levels of diets treatments (A(35.92 g•d-1), B(49.44 g•d-1), C(61.28 g•d-1), D (72.48 g•d-1)) and 4 20-day periods. pH, FAA-N (free amino acid nitrogen), NH3-N (NH3 nitrogen), PAA-N (peptide amino acid nitrogen), soluble protein N (nitrogen) and VFA (volatile fatty acid) were studied in different times before (07：30) and after feeding (10：00, 12：00, 14：00, 16：00) at last day. 【Result】 Ruminal pH was not affected by diets (P＞0.05). PAA-N, soluble protein N, FAA-N, total VFA and NH3-N were significantly differed among yaks with different diets (P＜0.05). NH3-N was also affected by diet×time interaction (P＜0.05). Acetate/propionate was not affected by different diets (P＞0.05). 【Conclusion】 Under the rations of the present study, when N of diet greater than 1.97%, most nitrogen fractions in yak do not response to the nitrogen level of the experimental diets. At 2.90% nitrogen of diet, acetate/propionate is the minimum and total VFA is the maximum.

Effect of Dietary Phosphorus Level on Performance and Phosphorus Excretion of Lactating Dairy Cows

ZHAO Heng-Ju, GAO Yan-Xia, LI Qiu-Feng, CAO Yu-Feng, LI Jian-Guo

Scientia Agricultura Sinica. 2011, 44(22): 4687-4693. doi:10.3864/j.issn.0578-1752.2011.22.016

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【Objective】 The aims of the present study were to investigate the effect of dietary phosphorus level on the performance and phosphorus excretion to environment in lactating cows. 【Method】 Thirty lactating Chinese Holstein cows were randomly allocated respectively into 3 treatments with 10 duplicates each. The phosphorus contents in the dietary of each treatment were 0.32%, 0.44% or 0.56% (dry matter basis), respectively. The trial lasted 75 days with 15-d adaptation period and 60-d experimental period. Samples were collected in the last 5 days. 【Result】Dietary phosphorus level of dairy cows showed no effects on the dry matter intake, milk yield and milk composition (P＞0.05). The contents of calcium, phosphorus, alkaline phosphatase and hydroxyproline in the serum and the saliva phosphorus content were all in the normal range. Daily phosphorus excretion to the environment through manure and urine of group fed diet with 0.32% P was decreased by 20.40 g and 45.84 g compared with the group fed diet with 0.44% P and 0.56% P, respectively. Fecal water-soluble phosphorus emissions from dairy cow fed diet with 0.32% P was respectively decreased by 15.24 g (P＜0.01) and 45.61 g (P＜0.01) compared with that fed diet with 0.44% P and 0.56% P. The daily feeding cost was respectively decreased by 0.60 yuan/cow and 0.40 yuan/cow. 【Conclusion】Therefore, dietary phosphorus level was 0.32% (dry matter basis) not only had no negative effect on the performance, but also significantlly decreased the fecal phosphorus excretion and improved economic benefit and ecological benefit of feeding. The recommended dietary allowance of phosphorus is 0.32%-0.44%.

Detection of Quantitative Trait Loci for Live Weight at Weaning in an Erhualian×Shaziling Population Using Whole-Genome High-Density SNP Markers

HE Yu-Yong, LI Wan-Bo, ZHANG Zhi-Yan, YANG Ming, 欧Yang-Jing , YANG Jie, REN Jun, XIAO Shi-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4694-4699. doi:10.3864/j.issn.0578-1752.2011.22.017

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【Objective】An Erhualian×Shaziling F2 resource population was constructed to map quantitative trait loci (QTL) for liveweight of piglets at the weaning age of 45 days and identify positional candidate genes related to weaned liveweight, providing a start pointing to detect the causal genes in the future.【Method】The F2 resource population was constructed and genomic DNA of all weaned piglets was genotyped with Illumina porcine 60 k DNA chip. Whole-genome linkage analysis was conducted to detect QTL for liveweight of piglets weaned at day 45. Positional candidate genes were characterized from Ensemble (EMBL-EBI) and NCBI (National Center for Biotechnology Information) database.【Result】A total of 3 QTLs were detected for the measured phenotype including one 5% genome-wide QTL on sus scrofa chromosome 2 (SSC2) and one 1% genome-wide QTL on sus scrofa chromosome 5 (SSC5) and sus scrofa chromosome 14 (SSC14), respectively. Five candidate genes were identified, including CYP2R1, COPB1 and PDE3B on SSC2 and NOP2, GDF3 on SSC5. 【Conclusion】 The QTL for liveweight of piglets weaned at day 45 were mapped on SSC2, SSC5 and SSC14, and 5 positional candidate genes were identified.

Molecular Cloning, Sequence Analysis and Tissue Expression of Follistatin Gene in Dazu Black Goat

LUO Yan-Mei, ZHANG Chao, ZHAO Zhong-Quan, FAN Jing-Sheng, ZHANG Jia-Hua

Scientia Agricultura Sinica. 2011, 44(22): 4700-4705. doi:10.3864/j.issn.0578-1752.2011.22.018

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【Objective】 The objective of this experiment is to clone the sequence of Dazu black goat follistatin gene and explore the expression levels of follistatin gene in different tissues.【Method】The cDNA of follistatin gene was cloned from total RNA of ovary tissue in Dazu Black Goat by RT-PCR. The expression analysis of follistatin gene was predicted with some related software tools,and the expression of follistin gene in different tissues was analyzed with real-time PCR.【Result】Sequence analysis indicated that the cloned follistatin gene cDNA was 1 217 bp in length (HM138665) containing an entire open reading frame (ORF), and encoding 344 amino acids. According to the Blast result in GenBank database, the cDNA nucleotides share 98% homology with Bos taurus. Real-time PCR analysis showed that the follistatin gene was expressed in spleen, lung, kidney, liver, heart, ovary and pituitary. The expression levels of follistatin gene were higher in kidney and lower in pituitary. The expression level of follistatin gene in kidney and heart were significantly higher than in pituitary (P＜0.05). No significant difference was found between the other tissues (P＞0.05).【Conclusion】The sequence of follistatin gene cDNA was obtained. And the follistatin mRNA expression was the hingest in kidney.

Effects of Deep-Litter Systems on Growth Performance, Biochemical Parameters and Swine House Environment in Weaned Piglets in Summer

FENG You, ZHANG Xiang-Bin, CHEN Xue-Ling, HUANG Zhong-Qiang, XIAO Jin-Song, WANG Xiu-Qi

Scientia Agricultura Sinica. 2011, 44(22): 4706-4713. doi:10.3864/j.issn.0578-1752.2011.22.019

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【Objective】The objective of this study was to investigate the effects of deep-litter systems on growth performance, biochemical parameters and swine house in weaned piglets in summer. 【Method】 A total of 300 piglets weaned at (28±3) days of age were randomly allocated into 2 treatments: traditional concrete-based house (TCBH) and deep-litter house (DLH). There were five replicates for each treatment and thirty piglets in each replicate. The experiment was lasted for 42 days. Body weights and feed intakes were measured every 14 days through the study. Temperature and relative humidity were measured every day. Blood samples were collected from four piglets from each replicate for biochemical parameters assay on the 15th, 29th and 43rd days. Air samples were collected for determination of the ammonia concentration in house from 29th to 35th days and from 36th to 42nd days. 【Result】 Average daily gain (ADG) of piglets in DLH group was higher (P＜0.05) compared with the TCBH group. Diarrhea ratio of piglets in DLH group was lower than those of TCBH group (P＜0.05) from 1st to 14th, 1st to 28th and 1st to 42nd days. There was no significant difference in average daily feed intake (ADFI) between the TCBH group and DLH group. Ammonia concentration in DLH group was lower (P＜0.05) compared with the TCBH group at 7:00 clock during the experiment days. There was no significant difference in air temperature and relative humidity between the TCBH group and DLH group (P＞0.05) .On the 14th and 28th days, the activity of alanine transarninase (ALT) in serum of DLH group was lower than that of TCBH group (P＜0.05), there was no significant difference in glutamic-oxal(o)acetic transaminase (AST) and alkaline phosphatase (ALP) activities in serum between the TCBH group and DLH group (P＞0.05). The content of glucose (GLU) and triglycerides (TG) in serum of DLH group was lower than that of TCBH group (P＜0.05) on 15th, 29th, and 43rd days.【Conclusion】The results demonstrated that deep-litter systems could decrease the activity ALT and decrease the content of TG and GLU in serum in summer. Deep-litter systems, which had no negative effect on growth performance of piglets and swine house environment, was suitable for weaned piglets production under the conditions of this experiment.

Optimal Protein Levels Required and Their Effects on Larval Antioxidation of Apis mellifera ligustica Spinola

LI Cheng-Cheng, YANG Wei-Ren, XU Bao-Hua, FENG Qian-Qian

Scientia Agricultura Sinica. 2011, 44(22): 4714-4720. doi:10.3864/j.issn.0578-1752.2011.22.020

Abstract ( 640 )

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【Objective】 The objective of this study is to research the effect of dietary crude protein levels on growth and larval antioxidation of Apis mellifera ligustica Spinola. 【Method】 Thirty cages of honey bees obtained from a local apiary were used in the study. Honey bees were randomly allocated into 6 groups (with 5 replications each and 1 cage in each replication). One of the groups was fed pure rape pollen only (control, group A) and others were fed the diets with crude protein levels at 15% (group B), 20% (group C), 25% (group D), 30% (group E) and 35% (group F), respectively.【Result】During the trial period, there was no significant effect of dietary crude protein on the population of bee colony (P＞0.05). The hatchability and percentage of pupation of group F were significantly higher than those of group B (P＜0.05). The larval protein of group E was significantly higher than that of groups A, B and C (P＜0.05). With the dietary protein level increasing, there was no significant effect on T-SOD activities of larva (P＞0.05), while the content of MDA was decreased (P＜0.05). 【Conclusion】These results indicated that the population of colony did not differ significantly with the crude protein content in this experiment, and 30%-35% of dietary crude protein level was optimal to maintain the highest growth performance and antioxidant activities.

Resistance to Antibiotics and Distribution of Tetracycline Resistance Determinants in Haemophilus parasuis from Pigs in South China

XU Cheng-Gang, GUO Li-Li, ZHANG Jian-Min, ZHANG Bin, LI Ang, CHEN Ji-Cheng, LI Jing-Yi, LIAO Ming

Scientia Agricultura Sinica. 2011, 44(22): 4721-4727. doi:10.3864/j.issn.0578-1752.2011.22.021

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【Objective】 Haemophilus parasuis is one of the most frequent pathogenic bacteria which has caused a serious of problems in pig industry all over the world. Therefore, it is important to evaluate the resistance to antibiotics and the distribution of tetracycline resistance determinants in H. parasuis from pigs in order to prevent and control this diseases. 【Method】 All H. parasuis were isolated from clinically diseased pigs and antimicrobial susceptibility to 21 antibiotics was performed using the broth microdilution method, and PCR was used to detect the presence of 9 tetracycline resistance genes. Then genetic relationship of the isolates which had tetracycline resistance was analyzed by Pulsed-?eld gel electrophoresis (PFGE). 【Result】 Most of the H. parasuis isolates showed resistance to many antimicrobial agents commonly used in veterinary clinic such as sulfamethoxazole, ?uoroquinolones, tetracycline, and gentamicin. Especially, more than 80% of the isolates were resistant to trimethoprim and sulfamethoxazole. Cephalosporins showed a good activity to H. parasuis isolates. Overall, the antimicrobial resistance of H. parasuis isolated from South China between 2008 and 2010 has been rising. All isolates showed multiple-drug resistance, resistance to 7 and 8 antibiotics was the main trend, the percentage were 19.13% and 17.39%. Tetracycline resistance gene tetB was widespread in H. parasuis. Analysis of PFGE typing implied that the isolates represented a wide variety of chromosomal backgrounds. 【Conclusion】 The study showed high resistance to antibiotics in H. parasuis isolated from pigs in South China, and tetB gene was widespread. The efflux of the drugs may be the major resistance mechanism of H. parasuis to tetracycline. This study helps to grasp the trend of drug resistance, and reveals mechanism of bacteria resistance to tetracycline.

Study on the Biocontrol Effects of Microbial-Fermentation Bed on the Pig Pathogen Escherichia coli in the Piggery

ZHENG Xue-Fang, LIU Bo, 蓝Jiang-Lin , SU Ming-Xing, LU Shu-Xian, ZHU Chang-Xiong

Scientia Agricultura Sinica. 2011, 44(22): 4728-4739. doi:10.3864/j.issn.0578-1752.2011.22.022

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【Objective】 The present paper dealt with the distribution of Escherichia coli and it’s virulent genes in the stroma cushion of piggery to explain the biocontrol effects of microbial-fermentation bed on the pig pathogen E. coli.【Method】The bacterium, E. coli isolated from 4 layers in the stroma cushion of piggery in different pig raising periods was analyzed by the method of eosin methylene blue (EMB) medium. The PCR detection was used to conform the species of E. coli by amplifying UdiA gene DNA fragments. The types of virulent genes were detected by the Multiplex–PCR. The simulation models were constructed to analyze the E. coli distribution in the microbial- fermentation bed. 【Result】 The results showed that 419 isolates were detected to be E. coli among 433 samples taken, of which, 59 isolates carried the special virulent genes. A total of 8 types of virulent genes were found, including estB, estA, elt, faeG, fedA, aidA–I, Stx2e and sepA. In the samples,the amount of E. coli had the highest positive rate (22.47%) in the one-month-used microbial-fermentation bed compared to that the lowest positive rate (4.23%) in the nine-month-used microbial-fermentation bed. The population dynamics of E. coli in each layer decreased as the time increased. For the population distribution in different layers, the pathogen E. coli was occupied least in the layer 2, less in the layer 1 or 3, and largest in the layer 4. The virulent genes distribution of E.coli was the similar trend to the population distribution in the piggery. From the dymamic models of E. coli population, it could be seen that the model was y=169.67x-1.0137 in the layer 1, y=0.1006x3-2.3733x2+16.094x-22.454 in the layer 2, y=313.11x-2.1885 in the layer 3, as well as y=0.3159 x3+6.0913x2- 35.634x+ 79.513 in the layer 4, from which, it displayed that the cushion stroma of piggery could inhibit the growth of E. coli by evidence of E. coli in the nine-month-used stroma cushion being much more less than that in the one-month-used stroma cushion with the decreasing rate ranged from 67.45% to 96.53%. It indicated that the microbial-fermentation bed could play an important role in biogicontrol of E.coli in the piggery.【Conclusion】The microbial-fermentation bed could restrain the growth of E.coli, especially for the bacteria carried with the virulent genes. The longer the microbial-fermentation bed was used the stronger the biocontrol effect was on the population of E.coli.

Screening of the Interaction Proteins of TaDREB6 Transcription Factor in Wheat

SUN Hai-Tao, XU Zhao-Shi, HOU Jian-Hua, YU Zhuo, ZHAO Yue, LI Lian-Cheng, CHEN Ming, MA You-Zhi

Scientia Agricultura Sinica. 2011, 44(22): 4740-4747. doi:10.3864/j.issn.0578-1752.2011.22.023

Abstract ( 822 )

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【Objective】TaDREB6 was used as a bait to screen the interaction proteins from wheat cDNA library by yeast-two hybrid system to further explore the DREB-mediated stress resistance mechanism in wheat.【Method】The TaDREB6 gene was amplified using wheat cDNA as the template. The wheat cDNA library and bait vector pGBKT7-TaDREB6 were constructed, respectively. The mixture of bait vectors pGBKT7-TaDREB6, pGADT7 and wheat cDNA library was introduced into yeast competent cells AH109 and cultured on SD/-Ade/-His/-Leu/-Trp plates for incubation at 30℃ for 3-5 days. Then the clones whose diameter was greater than 2 mm were selected for incubation on the plates containing X-α-gal to screen blue clones.【Result】A total of 102 candidate positive clones were sequenced and analyzed through homology analysis using the BLAST in NCBI. Homology analysis showed that those candidate proteins were related to energy metabolism, stress and defense, transport, transcription and translation, signal transduction, growth and development.【Conclusion】Function prediction of the candidate proteins suggested that TaDREB6 was possibly involved in several stress signal transduction pathways and play a important role in regulation stress resistance.

Research on the Relationship Between the Accuracy of SPAD Value and Sample Number in Growing Period of Cotton

CHEN Bao-Yan, MA Xing-Wang, YANG Tao, YAO Qing-Qing, XUE Xiang-Rong, WANG Bin, NIU Xin-Xiang

Scientia Agricultura Sinica. 2011, 44(22): 4748-4755. doi:10.3864/j.issn.0578-1752.2011.22.024

Abstract ( 720 )

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【Objective】 Research of the effects of observed samples of high yield drip irrigation cotton on N nutrition diagnosed by SPAD, to provide a theoretical basis for guiding fertilization of cotton with SPAD method. 【Method】 Adopting the method of drip irrigation under mulching in field, when N fertilizer rate was at 390 kg•hm-2, 5 programs for N fertilizer application were designed in order to supervise the 60 samples’ SPAD values of 5 N fertilizer application randomly during the whole growing period of cotton. The changing relationship between sample number and standard deviation under N fertilizer application rate of high-yielding cotton was analyzed after the yield was measured, and the influence of sample number on the accuracy of SPAD value in the growing period of cotton was defined. 【Result】 Differences were observed between the coefficient of variation of SPAD value and standard deviation; the power function relations of sample number of SPAD value (y) and standard deviation (x) at the stages of bud, early blooming, fill blooming, blooming and boll-forming, full boll-forming and early wadding are as follows: y=5632.2x-4.893（R2=0.7968），y=26456x-6.0437（R2=0.8732），y=50184x-6.5227（R2=0.9782），y=174890x-7.2601（R2=0.9655），y=27410x-5.743（R2=0.9182），y=16237x-5.3756（R2=0.942）, and y=ax-b(a,b parameters) model could be used to predict the variation of SPAD value as well as sample number in every growing period. Linear dependence of SPAD value and N content of leaves is 60＞30＞5. 【Conclusion】 The sample number influences the accuracy of SPAD average value, the larger the sample number the more accurate the SPAD average value, and the pertinence between SPAD average value and N content of the leaves will be better. Sample number of SPAD value and standard deviation is a power functional relationship.

Construction of a Hair-Follicle-Cell-Specific Expression Vector of Goat VEGF164 Gene and Its Transfection into Caprine Fetal Fibroblasts Cells Stably

BAO Wen-Lei, LI Bin, HOU Xin, LIU Jun-E, GUO Xu-Dong, WANG Zhi-Gang, LIU Dong-Jun

Scientia Agricultura Sinica. 2011, 44(22): 4756-4762. doi:10.3864/j.issn.0578-1752.2011.22.025

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【Objective】 The present study aims at constructing a eukaryotic expression vector pCDsRed2-KV of goat VEGF164 (vascular endothelial cell growth factor) gene and then to transfer it into Inner Mongolia Cashmere Goat (Capra hircus) fetal fibroblast (GFb) cells to obtain a transgenic cell clones, which stably expresses red fluorescence and expresses VEGF164 in hair follicle cells specifically. The transgenic cell clones can be used for nuclear transplantation. 【Method】 pCDsRed2-KV (6.3 kb), a hair-follicle-specific expression vector of VEGF164, was constructed by connecting VEGF164 gene to downstream of KAP6-1 promoter, and then inserting the KAP6-1 promoter-VEGF164 gene fragment into the basic vector pCDsRed2, which contains a DsRed expression unit. The Inner Mongolia Cashmere Goat fetal fibroblast (GFb) cells were transfected with the expression vector by lipofectamineTM2000. Transgenic cell clones were obtained after screening by G418. The recombinant of exogenous DNA was identified by polymerase chain reaction. 【Result】 The sequencing result showed that the VEGF164 gene was connected properly to the downstream of pKAP6-1, then the CMV promoter and the DsRed2 gene in sequence. Exogenous DNA in the cell clones was examined by PCR and the promoter KAP6-1 as well as VEGF164 gene has been integrated into GFb cells genome stably. 【Conclusion】 A hair-follicle-cell-specific expression vector of VEGF164 gene was constructed successfully and transfered into GFb cells. These data provide a way to obtain the transgenic goat by nuclear transfer in the future.

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