Scientia Agricultura Sinica

Mapping of Quantitative Trait Loci for Heading Date and Plant Height in Two Populations of Indica Rice

ZHANG Zhen-Hua, GUO Liang, ZHU Yu-Jun, FAN Ye-Yang, ZHUANG Jie-Yun

Scientia Agricultura Sinica. 2011, 44(15): 3069-3077. doi:10.3864/j.issn.0578-1752.2011.15.001

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【Objective】 Analysis of quantitative trait loci (QTLs) for heading date, plant height and yield traits and identification of QTLs having pleiotropism on plant height and yield traits but insignificant effect on heading date would facilitate the rice improvement for high-yielding potential. 【Method】 Two recombinant inbred line populations of indica rice were constructed using restorer line Milyang 46 as the common male parent and maintainer lines Xieqingzao B and Zhenshan 97B as the female parents, respectively. The two populations were planted in a trial site in multiple years for the detection of QTLs for heading date and plant height. 【Result】 A total of 12 QTLs for heading date and 11 QTLs for plant height were detected, among which only two QTLs for heading date locating on the short arm of chromosome 6 and the pericentromeric region of chromosome 7 were found in both populations. In comparison with QTLs for yield traits previously detected in the two populations, six QTL regions showing pleiotropic effects were found. Included are one controlling heading date, plant height and yield traits, three controlling heading date and yield traits, and two controlling plant height and yield traits. 【Conclusion】 In related to the common parent Milyang 46, the maintainer lines Xieqingzao B for dwarf-abortive cytoplasmic male sterility and Zhenshan 97B for wild-abortive cytoplasmic male sterility differ greatly in the genetic control of heading date and plant height, especially in plant height. QTL qPH2 in the interval RM6-RM240 on the long arm of chromosome 2 is of great potential in practicing the “higher in dwarf” approach to increase rice yield, since it showed a consistent effect on plant height and yield traits with insignificant effect on heading date.

Dynamic QTL Mapping of Wheat Protein Content in Developing Grains

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

Scientia Agricultura Sinica. 2011, 44(15): 3078-3085. doi:10.3864/j.issn.0578-1752.2011.15.002

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【Objective】Grain protein content (GPC) is an important quality factor in bread wheat, which is determined by quantitative trait loci (QTLs). In this study, to attain more genetic information of GPC accumulation，the number and genetic effects of QTLs/genes related to GPC were detected at different seed filling stages. 【Method】 QTLs/genes related to GPC were studied at five seed filling stages in six different environments, using a doubled haploid (DH) population derived from a cross between two elite Chinese wheat cultivars Huapei 3×Yumai 57. QTL mapping for developmental behavior of GPC was studied by unconditional and conditional quantitative trait locus (QTL) analyses in a mixed linear model.【Result】A total of nine unconditional QTLs and ten conditional QTLs were detected at five seed filling periods. The unconditional QTL named QGpc3A persistently expressed at all seed filling stages may play a most important role for the accumulation of GPC, the rest unconditional QTLs and conditional QTLs were only detected at a few or a special periods. At the period of 12 days after anthesis the gene expression was active, and the QTLs detected could account for 42.62% of phenotypic variation totally. However, the QTLs detected could totally account for only 17.43% of phenotypic variation at 22 d after anthesis with lowest phenotypic value. 【Conclusion】The dynamic change of GPC show a high - low - high trend and QTLs controlling GPC were developmental stage speci?c.

Assessment of Substitution Lines and Identification of QTL Related to Fiber Yield and Quality Traits in BC4F2 and BC4F3 Populations from G.hirsutum ×G.barbadense

LAN Meng-Jiao, YANG Ze-Mao, SHI Yu-Zhen, GE Rui-Hua, LI Ai-Guo, ZHANG Bao-Cai, LI Jun-Wen, SHANG Hai-Hong, LIU Ai-Ying, WANG Tao, YUAN You-Lu

Scientia Agricultura Sinica. 2011, 44(15): 3086-3097 . doi:10.3864/j.issn.0578-1752.2011.15.003

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【Objective】 A total of 276 SSR makers were used to assess the substitution lines and identify the QTLs related to fiber yield and quality traits in BC4F2 and BC4F3 populations from Gossypium hirsutum ×Gossypium barbadense with Gossypium hirsutum as recurrent parent, which could lay a foundation for further screening SSSLs (single segment substitution lines) , finely mapping QTL of fiber yield and quality-related traits and realizing molecular assistant breeding. 【Method】 GGT32 was employed to analyze the genotype of BC4F2 and BC4F3 individuals. Possible QTL were identified by one-way analysis of variance on SAS PROC GLM. 【Result】 A total of 50 SSSLs were detected in BC4F2 and BC4F3 individual populations, in which 9 plants contained the single homozygosis chromosome segment of Gossypium barbadense. Twelve plants were screened out with excellent fiber quality trait and a few chromosome segments from Gossypium barbadense. Fifteen QTLs for fiber yield-related traits and 19 QTLs for fiber quality-related traits were mapped in twelve linkage groups, which accounted for 2.80%-14.13% of phenotypic variance. 【Conclusion】 This study indicated that 4 QTLs related to fiber length could be detected in the two populations and 1 QTL for fiber length was reported in previous research, pleiotropism or linked genes existed in some loci that controlled different traits, and not all of additive genes were from the parent with excellent traits.

Effects of Sowing Date on Yield and Quality of DirectSeeding Rice of Different Types and Varieties

YAO Yi, HUO Zhong-Yang, ZHANG Hong-Cheng, XIA Yan, NI Xiao-Cheng, DAI Qi-Gen, XU Ke, WEI Hai-Yan, XIAO Yue-Cheng, WANG Xian

Scientia Agricultura Sinica. 2011, 44(15): 3098-3107. doi:10.3864/j.issn.0578-1752.2011.15.004

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【Objective】 The objective of the study is to elucidate the effect of sowing date on grain yield and its quality of direct seeding rice. 【Method】 Taking six representative rice cultivars of three types as experimental materials, the yield formation and main rice quality traits of direct seeding rice growing under the conditions of different sowing date were analyzed.【Result】The results showed that along with the delayed sowing date, the yields of three types of rice cultivars were all significantly decreased, and the changing degree was different; the yield decreasing caused by the decline of spikelets per panicle and filled grain percentage, panicles and 1000-grain weight had a little change. The law of changing of main direct seeding rice quality traits between different types under the conditions of different sowing date showed some similarities and differences. Along with the delayed sowing date, the appearance quality and cooking and eating quality had the same effect, but the trend was different, appearance quality became better while cooking and eating quality became worse. Milling quality and nutrient quality had different effects, milling quality of the type of medium-maturing and late-maturing medium japonica rice became better but worse the nutrient quality, milling quality of the type of early-maturing late japonica rice became worse but better the nutrient quality. 【Conclusion】 The direct seeding rice should be sowed as early as the time for harvesting the previous crop allows to attain high yield and improve cooking and eating quality, but reduce the appearance quality. Effects of sowing date on milling quality and nutrient quality were different because of types and varieties.

Effects of Rotational Tillage on Flag Leaf Photosynthetic Characteristics and Yield after Anthesis of Winter Wheat in Arid Areas of Southern Ningxia

HOU Xian-Qing, JIA Zhi-Kuan, HAN Qing-Fang, WANG Wei, DING Rui-Xia, NIE Jun-Feng, LI Yong-Ping

Scientia Agricultura Sinica. 2011, 44(15): 3108-3117. doi:10.3864/j.issn.0578-1752.2011.15.005

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【Objective】 The objective of the experiment is to seek the effects of no-tillage (NT)/Subsoiling (ST) rotational tillage system every other year on photosynthetic capacity and yield after anthesis of winter wheat.【Method】Based on three tillage treatments of NT/ST/NT, ST/NT/ST and conventional tillage (CT) in arid areas of southern Ningxia, it was investigated by measuring soil water, flag leaf area, chlorophyll relative content, photosynthetic characteristics, chlorophyll fluorescence parameters and yield under different soil tillage treatments after anthesis of wheat in 2010. 【Result】 Compared with conventional tillage (CK), the soil water storage in 0-200 cm soil layers of NT/ST/NT and ST/NT/ST was significantly increased by an average of 15.24% and 23.45% at flowering stage and filling stage of wheat, their flag leaf area and chlorophyll relative content were increased by an average of 12.78%, 8.89% and 25.20%, 11.19% (P＜0.05), respectively; their flag leaf daily average photosynthesis rate (P(——)n) was increased by an average of 1.55 and 1.68 μmol•m-2•s-1 (P＜0.05), their flag leaf daily average transpiration rate (T(——)r) was increased by an average of 0.44 and 0.58 mmol•m-2•s-1 (P＜0.05), but, their flag leaf daily average water use efficiency (WUE(————)) showed no obvious difference. At filling stage, two rotational tillage treatments had a higher level of maximal photochemical efficiency of PSⅡ in the darkness (0.52), actual photochemical efficiency of PSⅡin the light (0.40) and photosynthetic electron transport rate (92.67). Compared with conventional tillage, the different rotational tillage system achieved wheat grain yield increment of 6.88% and 11.98% (P＜0.05).【Conclusion】The different rotational tillage systems all showed better soil water conservation ability and high photosynthetic capacity after anthesis of wheat, significantly increased wheat yield under condition of drought and water shortage. So, the different rotational tillage system is a more suitable soil tillage system in arid areas of southern Ningxia.

Effect of Cadmium on Photosystem Activities of Maize (Zea mays L.) Leaves

LI Geng, ZHANG Shan-Ping, LIU Peng, GAO Hui-Yuan, WANG Jing-Feng, LIU Chen-Xu, DONG Shu-Ting, ZHANG Ji-Wang

Scientia Agricultura Sinica. 2011, 44(15): 3118-3126. doi:10.3864/j.issn.0578-1752.2011.15.006

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【Objective】 The effect of Cd on photosystem activities of maize leaves, and the action site and ways of Cd to photosynthesis were studied in order to provide a theoretical foundation for understanding of the injury mechanisms of Cd to the photosynthetic apparatus.【Method】Two maize varieties were chosen as materials. One is the common variety mainly planted in northern China, Zhengdan 958 (ZD958). Another is a new variety with higher yielding potential, Denghai 661 (DH661). Maize seedling was continuously cultured with different Hoagland’s solution containing 0, 10, 30 and 60 μmol•L-1 of CdSO4 after 3-leaf-stage. After 40 days of treatment, the parameters of gas exchange, chlorophyll a fluorescence transient and light absorbance at 820 nm of maize leaves were measured for analying the injury reason of Cd to the photosynthetic apparatus. 【Result】 It was found that the declines of Pn were not mainly related to stomatic factors under Cd stress. After Cd treatment, both fluorescence at K-step (Wk) of PSⅡ electron donor side and the fluorescence at J-step (Vj) of acceptor side were increased. However, the decrease of activity of electron donor side was lager than the electron acceptor side, which caused the decline of Ψo. With the increase of Cd concentration, the activity of RUBP (Ribulose 1,5-Biphosphate) decreased significantly, the utilization rate of NADPH was also decreased. They caused the electron accumulation in PSⅠ increased and the 820 nm light absorption of PSⅠ (ΔIR/Io) decrease. And then, which caused the activity declined of PSⅡ was not in keeping with PSⅠ, thus depressed the coordination between PSⅡ and PSⅠ (Φ(PSI/PSⅡ)).【Conclusion】Under the Cd stress, the main reason of Pn declined was not caused by non-stomata limitation. Cd decreased the activity of PSⅡ electron donor side, which resulted in the performance of PSⅡ declined and reduced the amount of electron transport to PSⅠ. The most importantly, the increased amount of electron accumulation in PSⅠ fed back by the carboxylation system was more than the decreased amount of electron transport form PSⅡ that resulted in the PSⅠ performance declined sharply, which depressed the coordination of PSⅡ and PSⅠ, and caused Pn declining.

Antimicrobial Action of an Endophytic Fungus from Sophora flavescens and Structure Identification of Its Active Constituents

HE Lu, JI Ming-Shan, WANG Yong, XU Hong-Bo, QI Bu-Kun

Scientia Agricultura Sinica. 2011, 44(15): 3127-3133 . doi:10.3864/j.issn.0578-1752.2011.15.007

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【Objective】The objective of this study is to isolate the endophytic fungi from Sophora flavescens and define its antimicrobial action. Also the identification of the category and structure of the active substance fermented from Sophora flavescens was conducted. 【Method】Tissue isolation and selective culture medium method was used to isolate the endophytic fungi. Plate method was applied to measure the antimicrobial characters. Eight kinds of solvent system was used to define its polar. Column chromatography and freeze drying, TLC biological autoradiography and HPLC were employed to isolate and purify, trace, and measure the purity, respectively. The structure was analyzed using 1H-NMR, 13C-NMR and LC-MS. 【Result】 An endophytic fungus strain was isolated from Sophora flavescens seed, numbered BS001. It inhibited the growth of Phytophthora capsici, Colletotrichum coccodes, Sclerotinia sclerotiorum, and Alternaria mali. The inhibition to Colletotrichum coccodes was the biggest with the antibacterial diameter of 2.734 cm. After isolating and purifying the fermentation liquor of BS001, the structure of bacteriostatic active composition was 6,7-(2′E)dibutenyl-5,8-dihydroxy-(Z)-cyclooct-2-ene-1,4-dione, which identified by spectroscopy. 【Conclusion】An endophytic fungus, BS001, which has inhibition to some pathogens, was got from Sophora flavescens. Also a new antimicrobial substance 6,7-(2′E)dibutenyl-5,8-dihydroxy-(Z)-cyclooct-2-ene-1,4-dione was extracted from fermented broth of BS001. This antmicrobial substance can be used for development of biopharmaceuticals and biopesticides.

Cloning and Sequence Analysis of a Full-Length cDNA of β-1, 3-Glucanase Gene from Musa pisangawake

DI Guo-Hui, RUAN Xiao-Lei, WU Li-Ting, TAN Xiao-Yong, LI Hua-Ping

Scientia Agricultura Sinica. 2011, 44(15): 3134-3141. doi:10.3864/j.issn.0578-1752.2011.15.008

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【Objective】 The objective of this study is to clone β-1, 3-glucanase gene which is a group of important emzymes in plant defenses against fungal diseases, and to research their resistant functions and effective applications.【Method】A novel DNA fragment encoding β-1, 3-glucanase was isolated from Musa pisangawake (AA) by applying a PCR strategy in which a pair of degenerated primers deduced from the two blocks with specific conserved amino acid regions in multiple plants was used. Then the full length cDNA (glu) of the fragment was obtained by rapid amplification of cDNA ends (RACE).【Result】The full length of glu is 1 114 bp. Compared with other sequences of β-1, 3-glucanase in GenBank, the deduced amino acid sequences of glu shared 54% -71% in homology.【Conclusion】 A novel β-1, 3-glucanase gene was cloned from the Musa pisangawake (AA). The result reported in this paper has laid a the foundation for further studies of β-1, 3-glucanase gene functions and development of transgenic plants to control plant diseases.

Prediction and Analysis of Verticillium dahliae VdLs.17 Secretome

TIAN Li, CHEN Jie-Yin, CHEN Xiang-Yong, WANG Jia-Ni, DAI Xiao-Feng

Scientia Agricultura Sinica. 2011, 44(15): 3142-3153 . doi:10.3864/j.issn.0578-1752.2011.15.009

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【Objective】The aim of this study was to predict and analyze secretome of Verticillium dahliae, an important soil-born fungal pathogen that causes vascular wilt diseases. 【Method】The softwares SignalP, TargetP, TMHMM, Big-pi and PROSITE were used to predict the sectetome of V. dahliae. Protein databases were used for secretome annotation to find putative pectinase, cellulose and pathogen-host interaction proteins. The proteins with common fungal effector character were investigated by a set of computer algorithms. By using BLASTP, the secretome of V. dahliae and V. albo-atrum was compared to find V. dahliae specifically secreted proteins. 【Result】A total of 922 possible secreted proteins were identified in V. dahliae scretome. The most frequent amino acid in signal peptides is alanine, and the least is glutamic acid and aspartic acid. The amino acid in the position -3 and -1 are very conserved. The secretome is equipped with 158 carbohydrate-active enzymes which include 10 pectic hydrolases and 14 pectic lyases, 190 putative pathogen-host interaction proteins, 97 RxLx[EDQ] motif containing proteins and 52 small cysteine-rich secreted proteins, and 58 V. dahliae specifically secreted proteins compared with V. albo-atrum. 【Conclusion】This study has provided secretome prediction algorithms for V. dahliae. Length of signal peptides vary greatly. Composition of signal peptides are mainly aliphatic amino acid and sequences of signal peptides near C-terminal are conservative. V. dahliae secretome are rich in potential pectinase, pathogen–host interaction protein, RxLx[EDQ] motif containing protein and small cysteine-rich protein which may play roles in V. dahliae pathogenetic mechanism.

Analysis of Effect of Soil Quality After Orchard Established in Hilly Loess Plateau

XUE Sha, LIU Guo-Bin, ZHANG Chao, ZHANG Chang-Sheng

Scientia Agricultura Sinica. 2011, 44(15): 3154-3161. doi:10.3864/j.issn.0578-1752.2011.15.010

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【Objective】 The study of soil quality after orchard established on slope cropland is of importance to exploration of the soil quality evolution and its evaluation during the agricultural production in loess hilly region.【Method】 Orchard at different years in loess hilly region was chosen as the subject to reveal changes in soil quality through experimental analysis and statistical method, and the slope cropland was used as reference. 【Result】 The results showed that soil physical properties and anti-erodibility did not change markedly or had a little decrease at the early stage of the land use from slope farmland to orchard, then improved significantly with the following years, and kept stable from 20 to 30 years. Soil organic C, total N content increased slowly with the years and reached the peak from 20 to 30 years. Available N improved significantly after 2 years and then increased gradually. Total P and available K dropped at the early stage of restoration and then increased, reached the peak from 20 to 30 years. Available P had no significant change in the first 10 years. The value of pH increased at first but then dropped, which was opposite to CaCO3. The content of soil microbial biomass C increased significantly at the early 5 years and then kept stable, soil microbial biomass P did not vary at the early 10 years and then increased markedly. Basal respiration increased drastically 10 years later and substrate-induced respiration increased significantly 5 years later, then kept stable. Metabolic quotient dropped markedly compared to slope cropland at the early stage but gradually improved with the increase of years, 15 years later, tended to be stable. Urease, alkaline phosphatase, saccharase and cellulase activity increased slowly at early stage and kept stable in 20-30 years. Catalase activity took on wave-like increase, amylase and polyphenol oxidase generally tended to slow drop with years and then kept stable from 10 to 30 years. Soil quality index (SQI), an integral indicator reflecting soil phyico-chemical and biological properties, increased markedly in the early 20 years and kept stable in 20-30 years. Regression analysis showed that SQI took on linear increasing with years (r=0.946). 【Conclusion】 The slope farmland had a low content of soil quality (including physico-chemical, biological properties) due to improper tillage practices and heavy erosion. Compared with slope cropland, soil quality improved quickly at the early stage of orchard and tended to be stable with the following years. The ecosystem of orchard faced a strong ecological stress because of its high output based on a high input.

Distribution Characteristics of Microbial Biomass Carbon and Nitrogen in Soil Aggregates under Tea Plantation

LIU Min-Ying, ZHENG Zi-Cheng, LI Ting-Xuan

Scientia Agricultura Sinica. 2011, 44(15): 3162-3168. doi:10.3864/j.issn.0578-1752.2011.15.011

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【Objective】 The distribution of microbial biomass carbon and microbial biomass nitrogen in soil aggregates of tea planted area was studied on purpose to reflect the influence of tea plantation on soil aggregates and its nutrient cycling, which can provide evidence for coordinating regional land use and returning farmland to forest/tea project. 【Method】 On the basis of field investigation and laboratory analysis, abandoned land and eucalyptus plantation were selected as contrasts, distribution features of microbial biomass carbon and microbial biomass nitrogen in soil aggregates under tea plantation were studied. 【Result】 The content of soil aggregates organic carbon comparatively increased as soil aggregates particles diameter reduced. The maximum soil aggregates concentrated with organic carbon were found at ＜0.25 mm diameter under tea plantation, abandoned land and eucalyptus plantation. The content of soil aggregates microbial biomass carbon, microbial biomass nitrogen comparatively decreased when soil aggregates particle diameter reduced under tea plantation, abandoned land and eucalyptus plantation. The maximum microbial biomass carbon, microbial biomass nitrogen was found in the soil aggregates of 2-5 mm diameter under tea plantation. Except ＜0.25 mm aggregates, the contents of soil aggregate microbial biomass carbon, microbial biomass nitrogen under tea plantation were higher than abandoned land and eucalyptus plantation within the same aggregates particle diameter. Soil aggregates microbial quotient generally decreased along with soil aggregates particles reduced under tea plantation, abandoned land and eucalyptus plantation. The maximum soil aggregates microbial quotient was mainly in the 2-5 mm aggregates under tea plantation, its distribution pattern was similar to microbial biomass carbon and nitrogen. 【Conclusion】The content of microbial biomass in tea plantation soil is more plentiful than in the contrast,especially for large aggregates,which shows that conversion of farmland to tea plantation is a relatively ideal model of set-aside patterns in study area.

Genetic Analysis of Gynoecious in Cucumber(Cucumis sativus L.)

LI Jian-Mei, QIN Zhi-Wei, ZHOU Xiu-Yan, XIN Ming, WU Tao

Scientia Agricultura Sinica. 2011, 44(15): 3169-3176. doi:10.3864/j.issn.0578-1752.2011.15.012

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【Objective】 The objective of this experiment was to analyze the genetic effects of gynoecious in cucumber in order to provide a theoretical basis of gynoecious and new varieties breeding. 【Method】 Gynoecious (D0401, D0420), subandroecious (D06103, D0819) and monoecious (HL-3) were used as test materials using Griffing Ⅳ1/2P (P-1) to build a hybrid combination. One was selected (D0420×D06103) to construct 6 generation groups, such as P1, P2, F1, F2, B1, B2. According to the investigations and statistics of the proportion of female flowers of individual plants within 25 knots, quantitative traits additive-dominant (A-D) models were used to analyze the female genetic regularity of cucumber in different seasons (spring, autumn), such as P1, P2, F1, F2, B1, B2. 【Result】The gynoecious in cucumber genetic consistent with the A-D genetic model. The addition variance and dominant-environmental variance were 51.05% and 19.66%, respectively. The dominant variance and addition-environmental variance were 0, narrow heritability, broad heritability was 51.05%, environmental-narrow heritability was 0, and the environmental-broad heritability was 19.65%. 【Conclusion】The gynoecious in cucumber is mainly a quantitative trait controlled by multiple genes; the proportion of female flowers is controlled by additive effect and conventional crossing breeding can be selected during the earlier generations. The environment (season) has a main effect on proportion of female flowers in cucumber.

Effects of Phenolic Acids on Mitochondria and the Activity of Antioxidant Enzymes in Roots of Seedlings of Malus hupehensis Rehd.

ZHANG Zhao-Bo, MAO Zhi-Quan, ZHU Shu-Hua

Scientia Agricultura Sinica. 2011, 44(15): 3177-3184 . doi:10.3864/j.issn.0578-1752.2011.15.013

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【Objective】Experiments were conducted to study the effects of ρ-hydroxybenzoic acid, phloroglucinol, syringic acid, benzoic acid, caffeic acid and ferulic acid on the functions of mitochondria and the activities of antioxidant enzymes in roots of Malus hupehensis seedlings which were apples common stocks.【Method】 Malus hupehensis seedlings were planted in bowl and were teated with 50 mL of 0.8, 4.0, 20.0 mmol•L-1 of the six kinds of phenolic acids at 3-day intervals. Fresh weight, activities of superoxidase (SOD), catalase (CAT) and peroxidase (POD), the opening of mitochondrial permeability transition pores (MPTP), membrane fluidity and cytochrome c/a were determined. 【Result】 Treatments with 0.8, 4.0, 20.0 mmol•L-1 of six kinds of phenolic acids all inhibited the growth of Malus hupehensis seedlings, reduced the activities of SOD, POD, and CAT, increased the opening of MPTP, decreased membrane fluidity and Cyt c/a, and impaired mitochondria in a sense. Benzoic acid at 20.0 mmol•L-1 had more significant inhibitions than other treatments. 【Conclusion】 The six kinds of phenolic acids at 0.8, 4.0, 20.0 mmol•L-1 could inhibit the activities of antioxidant enzymes and mitochondrial functions, and the inhibition depended on dose of phenolic acids. At the same concentrations, benzoic acid has stronger inhibition than other five phenolic acids.

Effects of Exogenous Salicylic Acid on Ions Contents and Net Photosynthetic Rate in Chrysanthemum Under Salt Stress

GUO Chun-Xiao, WANG Wen-Li, ZHENG Cheng-Shu, SHI Lian-Hui, SHU Huai-Rui

Scientia Agricultura Sinica. 2011, 44(15): 3185-3192. doi:10.3864/j.issn.0578-1752.2011.15.014

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【Objective】The aim of this experiment was to analyze the alleviation mechanism of exogenous SA on chrysanthemum under salt stress. 【Method】The effects of salicylic acid (SA) on contents of Na+, K+, Ca2+，Mg2+ in roots, leaves of chrysanthemum and net photosynthetic rate (Pn) were studied with spraying exogenous SA on leaves.【Result】The results showed that salt stress increased the content of Na+, but reduced the contents of K+, Ca2+, Mg2+ in roots and leaves of chrysanthemum. Exogenous SA treatment increased K+, Ca2+, Mg2+ contents by 37.70%, 16.44% and 20.54%, respectively, decreased Na+ content by 27.13% in the roots; increased the contents of K+, Ca2+, Mg2+ and Pn by 56.52%, 53.23%, 87.53% and 40.74%, respectively, decreased Na+ contents by 53.41% in leaves; increased the contents of K+, Ca2+, Mg2+ by 67.97%, 79.40% and 89.32%, respectively, decreased Na+ contents by 76.06% in the chrysanthemum at the 10th day after stress treatments. SK, Na, SCa, Na and SMg, Na were also significantly increased by exogenous SA under salt stress. Correlation analysis showed that the Pn has significant negative correlation with Na+ and significant positive correlations with K+, Ca2+, Mg2+. 【Conclusion】These results indicated that exogenous SA may alleviate the damage of chrysanthemum seedlings from salt stress via ion selective absorption and transportation.

The Core-Collection Construction of Flower Lotus Based on AFLP Molecular Markers

YANG Mei, FU Jie, XIANG Qiao-Yan, LIU Yan-Ling

Scientia Agricultura Sinica. 2011, 44(15): 3193-3205. doi:10.3864/j.issn.0578-1752.2011.15.015

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【Objective】 The construction of the core collection is very important for preservation, study and application of flower lotus (Nelumbo Adans.) germplasm resources. 【Method】 Based on AFLP markers, candidate core collections were constructed by using proportional strategy and UPGMA clustering sampling method within subgroups after dividing 395 flower lotus germplasm into 4 subgroups. By comparison of the genetic diversity parameters of these candidate core collection, such as the number of polymorphic loci, percentage of polymorphic loci, observed number of alleles, effective number of alleles, Nei’s gene diversity and Shannon’s information index, the core collection of flower lotus were screened, and genetic diversity comparison and t-test between original collection and core collection were analyzed. 【Result】 The core collection of flower lotus with 88 cultivars including 60 Chinese flower lotus (Nelumbo nucifera), 3 American louts (Nelumbo lutea), 16 hybrids between N. nucifera and N. lutea, and 9 Japanese lotus cultivars. The core collection reserved 22.27% samples of original collection. The reserved rate of number of polymorphic loci and percentage of polymorphic loci were 99.27%, and the reserved rate of observed number of alleles, effective number of alleles, Nei’s gene diversity and Shannon’s information index were 100.00%, 101.72%, 110.00% and 106.67%, respectively. t-test’s results showed that no significant difference was found in genetic diversity indexes between the core collection and original collection. 【Conclusion】 The core collection excluded the redundant germplasm from the original collection and conserved maximum genetic diversity with minimum lotus germplasm resource. These results demonstrated that the core collection could stand for original collection excellently.

Analysis of Glucomanno-Oligosaccharides by Fluorophore-Assisted Carbohydrate Electrophoresis

YAO Xi-Mei, SHI Bo, LIU Cheng-Cheng, LIANG Ping, LI Jing-Mei

Scientia Agricultura Sinica. 2011, 44(15): 3206-3213. doi:10.3864/j.issn.0578-1752.2011.15.016

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【Objective】 The objective of this study was to explore the conditions of FACE detecting glucomanno- oligosaccharides. 【Method】 Konjac glucomanno-oligosaccharides was used as the substrate, the optimal parameters were determined by investigating the effects of 3 kinds of fluorescer, the concentration of resolving gel, the conditions of derivatization reaction and sample impurities on FACE. 【Result】 The optimal conditions of FACE used in detecting glucomanno-oligosaccharides were as follows: fluorescer ANTS, concentration of resolving gel 25%, content of ANTS 2 μL, derivation reaction temperature 37℃ and derivation reaction time 16 h.【Conclusion】These findings show that FACE has a high resolution to glucomanno- oligosaccharides, and 14 kinds of components were detected by FACE, which provides an effective method for qualitative and quantitative analysis of glucomanno-oligosaccharides.

Advances in Research on Postmortem Tenderization Mechanism of Endogenous Proteolytic Enzymes in Muscle

HUANG Ming, HUANG Feng, HUANG Ji-Chao, XU Bao-Cai, ZHOU Guang-Hong, XU Xing-Lian

Scientia Agricultura Sinica. 2011, 44(15): 3214-3222. doi:10.3864/j.issn.0578-1752.2011.15.017

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The mechanism of postmortem tenderization of muscle has been the focus of meat science for years. It is generally accepted that improvement of meat tenderness during postmortem aging mainly results from limited degradation of myofibrillar proteins by endogenous proteolytic enzymes. Calpains are widely considered to be a major contributor, but not only ones to postmortem improvement of meat tenderness, which is the result of multi-enzymatic interaction. The roles of lysosomal cathepsins, proteasomes and calpains during postmortem tenderization of meat were briefly reviewed in the paper. The characteristics of apoptosis, activating pathways and caspases were introduced, meanwhile the potential contribution of caspases to postmortem tenderization was also discussed. In the last, the possibilities of interactions between caspases and calpains and contribution of both proteases together to meat tenderization were also analyzed in the review.

Effects of Adding Pectinase Produced by Penicillium Oxalicum from Goose to Diet on the Digestive Physiology and Immunological Mechanisms for Improving the Performance of Broilers

WANG Bao-Wei, JIANG Xiao-Xia, ZHANG Ming-Ai, GE Wen-Hua, YUE Bin

Scientia Agricultura Sinica. 2011, 44(15): 3223-3234. doi:10.3864/j.issn.0578-1752.2011.15.018

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【Objective】 The objective of the experiment is to study the mechanism of pectinase produced by penicillium oxalicum Currie and Thom from goose, to determine the adding effects and the usage of pectinase in broiler diet, and to provide a theoretical basis and a technical support for the application of pectinase. 【Method】 Two hundred and forty healthy 1-day-old broiler chickens under same condition were randomly divided into 4 groups. Group 1 was the control group fed normal level of basal diet The other three groups’ diets were added with, respectively, pectinase at 0.249%, cellulose at 0.168%, compound enzyme at 0.15% prepared with pectinase and cellulose in proportion of 1﹕1. 【Result】 The growth of broilers in pectinase group was significant, pectinase improved the digestibility of P, CF, NDF, ADF, Met, Lys, Cys, Thr, and Ile significantly (P＜0.05), and the digestibility of CP was improved by 17.31% remarkably (P＜0.01). On the 28th day, compared with the control group, pectinase group’s amylase, lipase, trypsin activities of duodenum and pancreas (P＜0.05), lipase activities of jejunum were significantly increased, pepsin activity was also higher than that of control group (P＜0.01). On the 49th day pectinase group’s amylase, lipase, trypsin activities of pancreas, amylase, trypsin activities of duodenum, lipase activity of jejunum and pepsin activity were significantly higher than that of the control group (P＜0.05), jejunum’s villous height was increased significantly (P＜0.05) and the thickness of intestinal wall was decreased significantly (P＜0.05). Pectinase group’s uric acid level and urea nitrogen level significantly decreased during 0-28 days and 29-49 days, blood glucose level significantly increased on 49th day (P＜0.05), the numbers of Escherichia coli and Salmonella significantly decreased (P＜0.05), the numbers of Lactobacillus and Bacillus bifidus significantly increased (P＜0.05). Serum antibody titres to NDV of 49-day-old significantly increased (P＜0.05). 【Conclusion】 The pectinase produced by penicillium oxalicum Currie and Thom from goose could increase the production of broiler performance and nutrient utilization, and improve the gut morphology, digestive enzyme activities and micro-ecological environment.

Localization and Differential Expression of LHR and LHR mRNA in the Oviduct of Jining Gray Goat During Estrous Cycle

LI Guang-Jun, WANG Hui, WANG Shu-Ying, HOU Yan-Meng, HUANG Li-Bo

Scientia Agricultura Sinica. 2011, 44(15): 3235-3245. doi:10.3864/j.issn.0578-1752.2011.15.019

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【Objective】 The objective of the experiment is to study the localization of LH receptor (LHR) and the expression of LHR mRNA in the oviduct during estrous cycle in Jining Gray Goat. 【Method】 The target fragment of LHR mRNA was retrieved, cloned and analyzed by using the real time fluorescence quantitative PCR, then the differences in expression were detected. At the same time, the immunohistochemical staining of LHR was done. 【Result】 The localizations of LHR immunoreactive substance in all parts of the oviduct at different stages of estrous cycle were carried out respectively, and it was found that LHR is mainly distributed on the epithelium of tubal mucosa, lamina propria and blood vessel endothelium. LHR mRNA expression was different in the oviduct of Jining Gray Goat at the four estrous cycle phrases. LHR mRNA in the linfundibulum was the highest during estrus (P＜0.01). In the ampolla, was the lowest during metestrus (P＜0.05). In the isthmus, was the highest during proestrus (P＜0.01). 【Conclusion】 The localization of LHR and its mRNA expression in the oviduct of goat reveals that LH has a kind of administration on the function of ovidct.

Expression, Purification and Antibacterial Activity Analysis of Rana dybowskii Antimicrobial Peptide Dybowskin-1ST

GUO Na, XIAO Xiang-Hong, XU Yi-Gang, CHAI Long-Hui, ZHANG Jing-Yu

Scientia Agricultura Sinica. 2011, 44(15): 3246-3251. doi:10.3864/j.issn.0578-1752.2011.15.020

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【Objective】 The aim of this study is to express Rana dybowskii antimicrobial peptide dybowskin-1ST via Pichia pastoris system and to analyze its antibiotic activity.【Method】The total RNA of Rana dybowskii skin was extracted by TRIzol method. The antimicrobial peptide dybowskin-1ST DNA were obtained by RT-PCR and cloned into expression vector pPIC9K, and recombinant pPIC9K-1ST was obtained. Then, the pPIC9K-1ST was electrotransformed into Pichia pastoris GS115, and the recombinant strain expressing dybowskin-1ST was generated. The recombinant strain was induced by the methanol and the interest protein was expressed via SDS-PAGE analysis. The interest protein was purified with Ni-agarose and its antimicrobial activity was detected using Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella enteritidis, Pseudomonas aeruginosa and Klebsiella pneumoniae.【Result】The recombinant Pichia Pastoris system expressing Rana dybowskii antimicrobial peptide dybowskin-1ST was well-constructed and the peptide dybowskin-1ST showed significant antimicrobial activity on gram-positive bacteria and gram-negative bacteria.【Conclusion】Rana dybowskii antimicrobial peptide dybowskin-1ST has a broad range of antimicrobial activity and a number of potential applications.

Progress in Research of FLP/FRT Site-Specific Recombination System in Higher Eukaryotes

ZHAO Ai-Chun, LONG Ding-Pei, TAN Bing, XU Long-Xia, XIANG Zhong-Huai

Scientia Agricultura Sinica. 2011, 44(15): 3252-3263. doi:10.3864/j.issn.0578-1752.2011.15.021

Abstract ( 1661 )

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FLP/FRT site-specific recombination system derived from 2 μm plasmid of yeast has been widely used in Arabidopsis thaliana, Oryza sativa Linnaeus, Mus musculus, Drosophila melanogaster, Caenorhabditis elegans and other higher eukaryotic organisms, and gradually become one of the powerful tools of genetic manipulation in transgenic animals and plants research areas. This review introduced the recombination principles of FLP/FRT system and its application in higher eukaryotes, and systematically summarized the main achievements of the system in transgenic plants, mammals, insects and other higher eukaryotic model organisms. In addition, the main problem, application prospect and developmental trend of the FLP/FRT system were discussed in this review.

Study on the Coexistence of GnRH, OT, SYN and S-100 Protein in the Same Luteal Cell of the Mid Phases of Oestrous Cycle in Karakul Sheep

ZHENG Mao-Liang, LI Guang

Scientia Agricultura Sinica. 2011, 44(15): 3264-3271. doi:10.3864/j.issn.0578-1752.2011.15.022

Abstract ( 577 )

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【Objective】The experiment was conducted to study coexistence of gonadotropin-releasing hormone (GnRH), oxytocin (OT), synaptophysin (SYN) and S-100 protein of dispersed or diffuse neuroendocrine system (dispersed or diffuse neuroendocrine system，DNES) in the same luteal cell. 【Method】 The corpus luteum was stained using pairwise combinations of OT, GnRH, SYN, and S-100 monoclonal antibody through double labeling immunofluorescence staining, respectively, at the mid phases of oestrous cycle of the Karakul sheep. 【Result】 Five kinds of immunofluorescence staining double positive cells combinations (OT and GnRH, OT and S-100, OT and SYN, GnRH and S-100, and GnRH and SYN) were respectively presented in luteal cell. In addition, the correlation analysis on 10 combinations of 5 kinds of double positive cells showed that there were 9 highly correlated combinations (r＞0.7). 【Conclusion】 This result reveals that there are DNES cells widespread in the same luteal cells，and there are OT and GnRH widespread in DNES cells. So, DNES cells may be the secretion cells of OT, and GnRH, and there are dual properties of both DNES cells and endocrine cells in luteal cells.

Effect on E. tenella Invasion into Host Cells in Vitro by Inhibiting Calcium Signal Transduction

ZHENG Ming-Xue, GU Shao-Peng, WANG Li-Xia, HAN Ke-Guang, LI Bao-Jun

Scientia Agricultura Sinica. 2011, 44(15): 3272-3278. doi:10.3864/j.issn.0578-1752.2011.15.023

Abstract ( 766 )

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【Objective】 The objective of the study is to investigate the relationship between Ca2+ signal transduction and Eimeria tenella sporozoite invasion, and to reveal the mechanism of coccidiosis (E. tenella). 【Method】 An assay system in vitro was used to study the effects of E. tenella sporozoites invasion into MDCK cell by extracellular Ca2+ deficiency or Ca2+ inflow blocker (nifedipine) or calmodulin inhibitor (trifluoperazine). Viability of MDCK cells and lactate dehydrogenase in supernatant liquid was examined. 【Result】 The inhibition rate of E. tenella sporozoite invasion increased with the decrease of the Ca2+ concentration. At low Ca2+ concentration of 600 μmol•L-1, the rate of E. tenella sporozoite invasion (23.33%) was significantly (P＜0.01) lower than that of the control group. When extracellular environment was absent of Ca2+ absolutely, the inhibition rate rose to 53.18%. Either nifedipine or trifluoperazine could inhibit the E. tenella sporozoite invasion obviously (P＜0.01), the inhibition rate of 71.41% and 97.13% were achieved by 10 μmol•L-1 nifedipine and 50 μmol•L-1 trifluoperazine, respectively. The inhibition rate of 98.59% was reached by nifedipine and trifluoperazine together. The viability of MDCK cells with E. tenella sporozoite invasion exceeded 90%, and the data was not remarkably different compared with the control group (P＞0.05). The same is as the viability of lactate dehydrogenase in supernatant liquid. 【Conclusion】 Extracellular Ca2+ deficiency, calciumion inflow blocker (nifedipine) and calmodulin inhibitor (trifluoperazine) could inhibit the E. tenella sporozoite invasion of MDCK cells. The host cell activity was not affected by E. tenella sporozoite invasion.

Study on China’s Swine Price Discovery Forming Mechanism ——Empirical Analysis on the Relationship Between Interregional Price

CHEN Yong-Fu, MA Guo-Ying, WU Bei-Bei, QIAN Xiao-Ping

Scientia Agricultura Sinica. 2011, 44(15): 3279-3279 . doi:10.3864/j.issn.0578-1752.2011.15.024

Abstract ( 918 )

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【Objective】 The objective of this study is to provide the transmission mechanism of swine price. 【Method】The common factors and information shares in major swine producing areas were measured by the permanent-transitory model and information share model based on the swine prices in these areas under the rising price volatility in China’s swine market. 【Result】The empirical results show that the swine prices in the three provinces of Sichuan, Hunan and Henan have an significant effect on China’s swine price formation and the rising swine price in Sichuan and Hunan plays a negative role in common factors, while in Henan has a positive effect. The permanent factor’s proportion of swine price in different provinces continues to show an upward trend generally, and the transitory factor’s proportion of swine price in Sichuan and Henan provinces is relatively high, as well as the swine price has a relatively high information share in market. 【Conclusion】 More attention should be paid to the stability of swine prices in Sichuan and Hunan. The flucluation of swine price in Henan should be minimized so as to reduce the magnitude of change in common factors. The epidemic and the downstream demand (pork demand) and other factors, which caused the change of the transitory factor’s proportion of swine price in Sichuan and Henan should be concerned so as to minimize the negative effects of disruption as far as possible.

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