Scientia Agricultura Sinica

Cloning and Characterization of Transcription Factor TaWRKY35 in Wheat (Triticum aestivum)

LIU Zi-cheng, MIAO Li-li, WANG Jing-yi, YANG De-long, MAO Xin-guo, JING Rui-lian

Scientia Agricultura Sinica. 2016, 49(12): 2245-2254. doi:10.3864/j.issn.0578-1752.2016.12.001

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【Objective】Abiotic stresses are major limitations to wheat production worldwide. Transcription factors play crucial roles in abiotic stress signaling in plants. It is predicted that there are at least 200 WRKY genes in common wheat genomes, yet only a few of them have been functionally characterized. The aim of this study is to decipher the roles of WRKY transcription factors in abiotic stress signaling and facilitate the utilization of WRKY genes in the improvement of abiotic stress tolerance in wheat.【Method】A wheat WRKY gene designated TaWRKY35 was cloned via the wheat full-length cDNA libraries. Quantitative real-time PCR was performed to identify the dynamic expression of target gene in different tissues at various developmental stages, and to characterize the transcriptional patterns responding to ABA, PEG, NaCl and low temperature treatments in wheat. To probe the subcellular location of TaWRKY35, the construct encoding TaWRYK35::GFP fusion protein was transferred into wheat protoplast by PEG mediated method. To characterize the function of TaWRYK35, target gene driven by the 35S promoter was delivered into Arabidopsis by Agrobacterium mediated method. 【Result】The cDNA of TaWRYK35 contains an 1 134 bp open reading frame, encoding a 377- amino acid protein. TaWRYK35 possesses a typical WRKY domain in the N-terminal and a C2HC type zinc finger domain in the C-terminal, belonging to group III of WRKY family. In 32 hexaploid wheat materials of highly polymorphic, TaWRKY35 coding region sequence is very conservative. The dynamic expression of TaWRKY35 was identified in different tissues at various developmental stages, and the highest expression occurred in the root base of seedling, while the lowest expression was observed in seedling leaf. Furthermore, its transcript was inducible by ABA, PEG, NaCl and low temperature treatments, yet the expression patterns to difference stress varied significantly. Subcellular localization indicated that TaWRKY35 specifically located in the nucleus. Overexpression of TaWRKY35 resulted in enhanced tolerance to high salinity, supported by improved cell membrane stability and survival rate relative to wild type Arabidopsis. 【Conclusion】TaWRKY35 transcription factor contains a WRKY and a C2HC type zinc finger domain, belonging to subgroup III of the WRKY family. The expression of TaWRKY35 occurs in different tissues at various developmental stages, and TaWRKY35 is an abiotic stress responsive gene. Overexpression of TaWRKY35 confers remarkably enhanced tolerance to high salinity.

Software Development of -ID Analysis for Crop Molecular Identity Construction

HU Zhen-bang, GAO Yun-lai, QI Zhao-ming, JIANG Hong-wei, LIU Chun-yan, XIN Da-wei, HU Guo-hua, PAN Xiao-cheng, CHEN Qing-shan

Scientia Agricultura Sinica. 2016, 49(12): 2255-2266. doi:10.3864/j.issn.0578-1752.2016.12.002

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【Objective】Based on the SSR molecular marker data of crops resources, software ID analysis was developed using visual Basic6.0 for crop molecular identity construction, which could screen primer combinations rapidly and accurately for efficient cultivar identification. 【Method】The crop molecular ID theory was proposed by Mr. Qingshan Chen. SSR markers in crop varieties showed high polymorphism characteristics, and a set of markers was permutated and combined to quickly and effectively divide varieties with step amplification method. Finally, 40 paires of SSR data of 40 soybean varieties in Heilongjiang province were used for identity construction with the software. 【Result】 ID analysis software was developed with core algorithm by using Visual Basic 6.0 to design man-machine interactive interface. This software has integrated full-library construction, partial-library construction, molecular ID determination, and database merging function. The full-library construction was the core functions together which could quickly obtained the minimum SSR prime combination to distinguish all the varieties. Partial-library construction could be used for some target varieties identification. For an unknown materials, with the already existing molecular identity database and the SSR analysis data, molecular ID determination could be used to determine the variety names or similar varieties. Database merging could be used to integrate several experimental data into a data set. The following results were analyzed by full-library construction with case data. First, among the 40 pairs of SSR data of 40 soybean cultivars, a total of 13 primers were excluded because missing data were too much and did not meet the standards, they were Sat_111, Satt218, Satt231, Satt685, Satt514, Satt551, Satt077, Satt358, Satt424, Satt100, Satt838 ,Satt893, and Satt891. Second, 6 primers were excluded because they showed high similarity coefficient with other primers, and they were Satt253, Satt192, Satt417, Sat_229, Satt127, and Satt496. Third, 5 varieties showed 7 specific alleles among all 40 varieties. They were, allele 3 of Satt516 and allele 1 of Satt253 showed in Dongnong36, allele 1 of Sat_229 showed in Nenfeng 17, allele 3 of Satt192 showed in Dongnong42, allele 1 of Satt206 and allele 4 of Satt244 showed in Beifeng 19, and allele 1 of Satt363 showed in Heihe14. So these specific allelic genes could directly identify the varieties. Forth, only seven pairs of SSR primers could distinguish 40 soybean varieties completely. The primer combinations were Satt398, Satt380, Satt453, Satt288, Satt244, Sat_092, and Satt206. 【Conclusion】 In this research, the software analysis ID was developed to construct crop molecular identity. The software has a friendly interface and easy to be used, high efficiency and flexible. The construction of the crop molecular identity can be realized completely by using a single software, and thus achieving the purpose of variety identification. With the development of technology of the application of capillary electrophoresis and the molecular identity theory, the development of an automatic molecular identity analysis system and even a rapid resource identification system will become possible.

Analysis of Root Absorption Characteristics and Nitrogen Utilization of Wheat Genotypes with Different N Efficiency

XIONG Shu-ping, WU Ke-yuan, WANG Xiao-chun, ZHANG Jie, DU Pan, WU Yi-xin, MA Xin-ming

Scientia Agricultura Sinica. 2016, 49(12): 2267-2279. doi:10.3864/j.issn.0578-1752.2016.12.003

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【Objective】This study was aimed at clarifying the physiological mechanisms of nitrogen uptake and utilization of wheat, and providing a theoretical basis for breeding and high-efficiency cultivation of wheat genotypes with high nitrogen efficiency, by analyzing the difference in root absorption characteristics and shoot nitrogen utilization of wheat genotypes with different nitrogen efficiencies. 【Method】 In this study, Nitrogen-efficient cultivars Zhoumai 27, Zhengmai 366 and Nitrogen-inefficient cultivars Zhoumai 28, Kaimai 20 were selected from 16 wheat genotypes in yield conditions. The relationship of root and shoot, root dry weight, root physiological activity, and the difference in nitrogen transport and distribution capability of root and shoot were researched under different nitrogen levels.【Result】The result showed that there were no significant differences in root characters of two types of wheat genotypes before jointing stage. After jointing stage, root dry weight, root to shoot ratio, root total absorption area and root active absorption area of Zhoumai 27, Zhengmai 366, and 28 were significantly higher than Kaimai 20. Root vigor of N-efficient genotypes was significantly higher than that of N-inefficient genotypes. The nitrogen accumulation amount at mature and nitrogen absorption amount after anthesis of Zhoumai 27, Zhengmai 366, and 28 were significantly higher than Kaimai 20. The grain yield, plant nitrogen use efficiency, nitrogen physiological efficiency, and nitrogen distribution ratio in grain of N-efficient genotypes Zhoumai 27, Zhengmai 366 were significantly higher than N-inefficient genotypes Zhoumai 28 and Kai Mai 20. The translocation amount to grain of N-efficient genotypes Zhoumai 27, Zhengmai 366 were significantly higher than those of N-inefficient genotypes Zhoumai 28, Kaimai 20, which vegetative organs accumulated nitrogen before anthesis. Compared with conventional nitrogen application rate, reduced supply nitrogen, root dry weight, root total absorption area, root active absorption area, root vigor, nitrogen accumulation amount at mature, nitrogen accumulation translocation to grain, and grain yield of the four wheat genotypes were decreased, when nitrogen application rate was reduced. At the same time, root to shoot ratio, nitrogen uptake efficiency, plant nitrogen use efficiency, and nitrogen physiological efficiency were increased when nitrogen application rate was reduced, root dry weight of four wheat genotypes, except for Zhoumai 28. At the same time, root total absorption area, root active absorption area, root vigor, nitrogen accumulation amount at mature, nitrogen accumulation translocation to grain, and grain yield of the four wheat genotypes were increased. And the root to shoot ratio, nitrogen uptake efficiency, plant nitrogen use efficiency, and nitrogen physiological efficiency were decreased.【Conclusion】The higher root biomass, root activity, root total absorption area, and root active absorption area of N-efficient wheat genotypes contributed to its absorption of nitrogen; it was the basis of nitrogen efficiency. The higher Nitrogen translocation, nitrogen grain distribution capacity, and reasonable shoot ratio of N-efficient wheat genotypes promotes its utilization of nitrogen, and it was the key for nitrogen efficiency. Although the N-inefficient wheat cultivar Zhoumai 28 had the strongest nitrogen absorption capacity, it had low transport capacity of nitrogen fertility and excessive shoot ratio after anthesis, which limited rational nitrogen utilization of wheat plant, which was not conducive to the improvement of nitrogen efficiency. The lower nitrogen efficiency of Kaimai 20 was due to its lower nitrogen absorption capacity.

Root Morphological Characteristics of Cotton Genotypes with Different Phosphorus Efficiency Under Phosphorus Stress

LUO Jia, HOU Yin-ying, CHENG Jun-hui, WANG Ning-ning, CHEN Bo-lang

Scientia Agricultura Sinica. 2016, 49(12): 2280-2289. doi:10.3864/j.issn.0578-1752.2016.12.004

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【Objective】Efficient genotype cotton plays important roles in promoting low phosphorus (P) stress from the perspective of the root morphology. The purpose of this study is to provide a scientific basis for identifying the main factors that affecting phosphorus absorption of cotton and improving efficiency of nutrient utilization of genetic improvement through root plasticity. 【Method】A pot experiment was carried out to test the effects of P supply levels (low P stress 0, Suitable P 150 kg·hm-2) with two P genotype （efficient XH18, CCRI-42, XLZ19; inefficient XLZ13, XLZ17）cottons on root morphology and the relationships between root morphology and P uptake. 【Result】 Cotton biomass and P uptake were significantly reduced by low P stress. Efficient genotype cotton biomass and P uptake were 1.21-2.08 and 1.35-1.91 times as much as those of inefficient genotype cotton, respectively. Phosphorus significantly increased Olsen-P content in soil. Olsen-P of P efficient genotype cotton was significantly decreased more than those of two P inefficient cottons compared with phosphorus suitable conditions under low P stress. Olsen-P in the 0-25cm layer of P efficient genotype cotton was less than P inefficient cotton, compared with XLZ13 and XLZ17 decreased by 21.1% and 30.1% in the low P stress environment. The total root length, total root surface area, total root volume and average root diameter of cotton were significantly reduced under low P stress. The total root length, total root surface area, total root volume of P efficient genotype cotton were 1.54-1.97, 1.52-1.92 and 1.47-1.84 times as much as that of those of P inefficient cotton. P efficient genotype cotton’s specific root length, specific root surface area and specific root volume were significantly higher than those of two P inefficient cottons under low P stress, were 1.10-1.25, 1.07-1.22 and 1.01-1.16 times as much as that those of P inefficient genotype cottons. The average root diameter of P efficient genotype cotton was significantly less than P inefficient cotton, which was 34.2%-70.2% of P inefficient genotype cotton in the low P stress environment. Principal component analysis showed that the total root length, total root surface area, total root volume, root dry mass, middle root length and coarse root length could be used to distinguish two P genotype cottons. General linear model variance decomposition results showed that the total root length, total root surface area, total root volume, root dry mass, middle root length and coarse root length made great contributions to P uptake of cotton. 【Conclusion】P efficient genotype cotton adapted to low P stress through greatly increasing the proportion of fine roots, reducing overall fineness root, prompting an increase of specific root length, and increasing construction efficiency of root.

Construction and Application of a Traceability System for the Vehicle Safety of Harmless Handling of Pigs Die of Diseases

GAO Xu, XIE Ju-fang, FAN Lei, HU Yi-nong, JIA Wei-ya, LI Xue

Scientia Agricultura Sinica. 2016, 49(12): 2290-2300. doi:10.3864/j.issn.0578-1752.2016.12.005

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【Objective】 The purpose of the study was to trace vehicles for the transport and harmless handling of pigs die of diseases from the harmless treatment plant to the circulation of safety traceability between collection points. 【Method】The system takes the transport vehicle for the harmless treatment of dead pigs as the research object. The system frame is designed with a three layer structure: a data service layer, a data processing layer, and a data collection layer. The data collection layer is composed of Beidou/GPS dual mode receiver, a temperature sensor, a wireless radio frequency reader, an active electronic label, and a GPRS wireless transmission module. The study sought to collect the transport vehicle location information, compartment temperature information, and disinfection point vehicle active electronic tag information of the traceability safety system of the transport vehicle for harmless handling of dead pigs, and then transfer this data. The location information of the transport vehicle is mainly gathered by use of the collected data of the Beidou navigation and positioning system. The GPS navigation and positioning data from the WGS-84 coordinate system was converted to the BJ-45 coordinate system of the Beidou navigation and positioning system, and the GPS navigation and positioning system was used to modify the data when the Beidou navigation positioning system data appeared to show a larger deviation, to achieve the purpose of improving the acquisition accuracy of the transport vehicle location information. The data processing layer was implemented to extract, modify, and store the collected data. The data service layer mainly provided information services for the staff and monitoring department. This system was developed by using C# language in a Studio Visual 2010 integration environment and using SQL language to store and modify the data in the database server SQL 2008. 【Result】 The function of the system includes route safety management, disinfection safety management, temperature monitoring, control management, health and epidemic prevention management, and safety pre-warning management. An ant colony algorithm was used to carry out the simulation of transport vehicle path planning for the harmless handling of dead pigs away from large farms, living groups, and other areas in the driving region. The simulation result was realistic, and it provides a reference for transport vehicles for the harmless handling of dead pigs to plan a reasonable path in the designated transport area. The traceability safety system of the transport vehicle for the harmless handling of dead pigs provides management staffs of harmless treatment plants with monitoring of transport vehicles, intelligent allocation of transport vehicles, compartment temperature monitoring, and vehicle information query functions. The system carried out on-site testing in a harmless handling of dead pigs pilot plant in Lianshui County, Jiangsu Province. The result showed that the hardware module of the system runs stably, the network packet loss rate was 0.26%, and the vehicle identification error rate was 0.97%. Through the ArcGIS monitoring and management function module, continuous monitoring of the transport vehicles running routes was done for 2 hours. Testing of the temperature monitoring management function, and the disinfection safety management function showed that each module of the system was working properly, and met the requirements of the traceability safety system of the transport vehicle for the harmless handling of dead pigs. Meanwhile, it can provide services for a monitoring and management department of animal health and epidemic prevention through the monitoring client by means of telephone, network, and short messages. It can also send the monitoring data such as vehicle running route, compartment temperature, and other information to the supervision department in real time to realize the implementation of the comprehensive supervision and management of a transport vehicle for the harmless treatment of dead pigs and ensure animal health and safety. 【Conclusion】This study has provided an effective method off safety management for a transport vehicle for the harmless treatment of dead pigs, accomplished the comprehensive supervision and management of transport vehicle for the harmless treatment of dead pigs, and reasonably avoided the occurrence of animal health and safety incidents. As it provides a reference for other harmless treatment of dead pigs die of diseases integrated management systems, it has practical value.

Cloning and Expression Analysis of Apoptosis-Related Gene FpTatD in Fusarium pseudograminearum

CHEN Lin-lin, HOU Ying, DING Sheng-li, SHI Yan, LI Hong-lian

Scientia Agricultura Sinica. 2016, 49(12): 2301-2309. doi:10.3864/j.issn.0578-1752.2016.12.006

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【Objective】The objective of this study is to investigate the potential biological functions of apoptosis-related genes FpTatD in Fusarium pseudograminearum, for this purpose, FpTatD genes were identified and cloned in F. pseudograminearum, and the expression of these genes were examined at mycelia, conidia and infection stages.【Method】 The known TatD proteins were obtained from GenBank, and four TatD candidates in fusariums were identified by BlastP. FpTatD genes and open reading frames (ORF) were amplified from genome DNA and cDNA by PCR, and the transcription levels in mycelia, conidia and infection were examined by qRT-PCR. The expression levels of FpTatD genes in the affinity interaction and non affinity interaction were assayed by RNA-seq.【Result】Four TatD candidates were identified in Fusariums, which divided into two significant branches by the phylogenetic tree. TatD1 and TatD2 belong to an ancient family that is conversed in almost all eukaryotic, while TatD3 and TatD4 seem to come from a unique TatD family that only exists in plants and fungi. Four FpTatD candidates, which identified in F. pseudograminearum were cloned, and the full-length sequences of FpTatD1, FpTatD2, FpTatD3 and FpTatD4 were 993, 1 331, 1 227 and 1 176 bp. FpTatD2 contains an intact open reading frame with 94 and 55 bp non-coding sequences in 5′ terminal, while FpTatD1, FpTatD3 and FpTatD4 consist continuous open reading frames. The four clones encode 36.37, 43.13, 45.59 and 44.25 kD proteins. All FpTatD contained the conserved DNase domain and most conserved amino acid residues. qRT-PCR analysis revealed that FpTatD1 and FpTatD2 were highly expressed in F. pseudograminearum, and both were highly up-regulated at early stages of infection. Especially, the expression level of FpTatD1 escalated to 8.8 and 7.6 times in 36 h and 3 d. However, FpTatD3 and FpTatD4 were poorly expressed at all stages. Thus, FpTatD1 and FpTatD2 might play important roles in F. pseudograminearum. RNA-seq analysis was consistent with qRT-PCR results that FpTatD1 and FpTatD2 were highly expressed and up-regulated at early stages of infection. In addition, Compared to the affinity interaction between F. pseudograminearum and GM301, FpTatD genes showed higher expression levels in the non affinity interaction between F. pseudograminearum and ZM24.【Conclusion】Apoptosis-related gene FpTatD might play an important role in the interaction between F. pseudograminearum and its host.

Effects of Extreme High Temperature on Survival Rate, Reproduction, Trehalose and Sorbitol of Frankliniella occidentalis

JIANG Shan, LI Shuai, ZHANG Bin, LI Hong-gang, WAN Fang-hao, ZHENG Chang-ying

Scientia Agricultura Sinica. 2016, 49(12): 2310-2321. doi:10.3864/j.issn.0578-1752.2016.12.007

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【Objective】The western flower thrips (Frankliniella occidentalis) is an important invasive and quarantine pest in China which created massive economic losses in vegetables and flowers. The objective of this study is to explore the influence of extreme high temperature (EHT) on survival rate, fecundity and content of trehalose and sorbital of F. occidentalis, and to provide a basis for the control of F. occidentalis. 【Method】After 2nd nymphs and adults exposed to 45℃ for 2 h twice (24 h interval under variable temperatures 22℃ (4 h)-25℃ (8 h)-28℃ (4 h)-25℃ (8 h) between two heat treatments), the survival rate, female adult longevity, offspring population dynamics, sex ratio and content of trehalose and sorbital were detected and analyzed. 【Result】In comparison with the control, the survival rate of each stage was less than 50% (0 for 3rd and 4th nymph, 41.38% for female adults, 5% for 1st nymphs and 21.36% for 2nd nymphs), parental and offspring female adult longevity and fecundity significantly reduced (P＜0.05), the content of trehalose in 2nd nymphs and adults significantly declined (P＜0.05), and the content of sorbital in 2nd nymphs and female adults significantly increased after heat-shocked on parental 2nd nymphs and female adults (P＜0.05). Meanwhile, higher female adult longevity and fecundity, lower content of trehalose, higher content of sorbital in F. occidentalis sufferred from twice 45℃ for 2 h heat shock on parental adults was expressed more than those in F. occidentalis sufferred from heat-shock on parental 2nd nymphs. Interestingly, the influence of twice 45℃ for 2 h heat shock not only occurred on parental generation but subsequent generations (F₁and F₂ generations), and the trend of F₁ and F₂ generations was the same as the parental generation.【Conclusion】Twice 45℃ for 2 h exposes on parental F. occidentalis significantly affected the development, reproduction and physiology of present and offspring generations. The changes of trehalose and sorbital content had a correlation with the population dynamics of F. occidentalis.

Study on Present Situation and Characteristics of Plow Pan in the Northern Region of Huang Huai Hai Plain

ZHAI Zhen, LI Yu-yi, PANG Huan-cheng, WANG Jing, ZHANG Li, DONG Guo-hao, GUO Jian-jun, GUO Zhi-hui

Scientia Agricultura Sinica. 2016, 49(12): 2322-2332. doi:10.3864/j.issn.0578-1752.2016.12.008

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【Objective】The objective of this paper is to study the present situation and characteristics of plow pan in the northern region of Huang Huai Hai plain.【Method】 In this study, 108 investigation points distributed in Wuqiao County of Hebei province and Ling County of Shandong province were selected according to the soil texture distribution data and the Second China National Soil Survey. At each investigation point, the soil moisture content, soil bulk density and penetration resistance of different soil levels from top surface to 45 cm during the jointing stage of winter wheat were determined. 【Result】 The average thickness of topsoil in the northern region of Huang Huai Hai plain was 14.74 cm, and the plow pan distributed from 15 to 30 cm existed at about 76% of all investigated points. It was also found that the bulk density and penetration resistance among each soil layer have significant differences. The bulk density of plow pan was about 1.54 g·cm^-3which was the largest in soil profile. Also the penetration resistance of plow pan during the jointing stage of winter wheat varied from 1 371.00 kPa to 4 256.00 kPa, which was significantly higher than that of top soil and subsoil. During the whole growth period of winter wheat, the penetration resistance of plow pan was always larger than 2 000 kPa which restrained the growth of root, and primarily distributed in shallow soil, and this led to the increased water stress to winter wheat and nutrient uptake from deep soil in water deficient area. A very significant correlation between soil penetration resistance and bulk density, soil water content was observed. In addition, the penetration resistance of soil showed a increasing trend with the increased soil bulk density and the regression equation is y=3 854.09x+3 891.99 (y is penetration resistance, x is the bulk density, r=0.84). When the bulk density is less than 1.4 g·cm^-3, soil penetration resistance was lower than 2 000 kPa, which was not an obstacle for root growth. When the soil bulk density is 1.4 g·cm^-3 or more, the effect of soil penetration resistance on winter wheat growth mainly depended on the soil moisture, and the regression equation between them is y= -75.93x + 3 153.83 (y is penetration resistance, x is the water content of soil, r= 0.82). 【Conclusion】 In a whole, plow pan is widespread under the traditional rotary-based tillage mode in the northern Huang Huai Hai, which would be an obstacle for crop root growth and the utilization of soil nutrients. So moderately breaking the plough pan and building a reasonable soil structure is required.

Effects of Two Biochars on Red Soil Aggregate Stability and Microbial Community

HE Yu-ting, WANG Chang-quan, SHEN Jie, LI Bin, LI Bing, CHEN Lin, PAN Xing-bing

Scientia Agricultura Sinica. 2016, 49(12): 2333-2342. doi:10.3864/j.issn.0578-1752.2016.12.009

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【Objective】 The objective of this paper is to study the influences of biochar derived from tobacco stems and mulberry branches on red soil aggregate stability and microbial community structure abundance, provide high quality modified materials for cultivating structural stability of soil aggregate. 【Method】 Soil samples were incubated for four months by adding four different rates (1%, 2%, 4% and 6%) of tobacco stems biochar (Y₁, Y₂, Y₄, and Y₆) and mulberry branches biochar (S₁, S₂, S₄, and S₆), and non-biochar control (CK). Then, sieving methods were used to detect the distribution of aggregates (＞2, 1-2, 0.5-1, 0.25-0.5, ＜0.25 mm) and stability (estimated by the mean weight diameter: MWD, ＞0.25 mm soil aggregates: R_0.25 and percentage of aggregate disruption: PAD). The numbers of soil fungi, bacteria and actinomycetes were measured by microbial dilution plate counting cultivation method. 【Result】 Water-stable aggregates were changed significantly after biochar additions. Compared with CK, Y₄and S₄ treatments significantly increased the 0.5-1 mm fractions by 61.0% and 43.6%; Y₁ and S₂ treatments significantly increased the 0.25-0.5 mm fractions by 40.8% and 27.1%, while the ＜0.25 mm aggregates reduced by 9.2% and 8.4%. Compared to CK, both Y₂ and S₂ enhanced the values of MWD more than 10% and Y₁ and S₂ treatments increased R_0.25by 31.4% and 28.7%, respectively. Accordingly, the PDA in the Y₆ and S₆ treatments were decreased by 22.0% and 18.2%, respectively. Furthermore, biochar additions significantly increased the soil fungi, actinomycetes and bacteria community. The Y₄ and S₄ treatments resulted in a richest amount of microbial community. In the tobacco stems treatments, a significant correlation existed between the aggregate stability index (MWD, R_0.25) with the fungi (R²=0.89, P=0.030; R²=0.86, P=0.039) and actinomycetes (R²=0.87, P=0.035; R²=0.90, P=0.021). Moreover, PAD significantly reduced with the increased fungi (P＜0.01), and actinomycetes and bacteria (P＜0.05).【Conclusion】The biochar derived from tobacco stems and mulberry branches enhanced the formation of macro-aggregate (0.25-1 mm), soil aggregates stability and abundance of soil microbial communities. The effect of tobacco stems biochar was better than that of mulberry branches, and the appropriate application is 2%-4%.

Effects of Irrigation Quota on Distribution of Soil Water-Salt and Yield of Spring Maize with Drip Irrigation Under Mulch

WANG Zeng-li, DONG Ping-guo, FAN Xiao-kang, WANG Tian-ren

Scientia Agricultura Sinica. 2016, 49(12): 2343-2352. doi:10.3864/j.issn.0578-1752.2016.12.010

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【Objective】 The objective of the experiment was to investigate the effects of irrigation quota on distribution of soil water-salt, yield and water use efficiency of spring maize under the condition of drip irrigation under film. 【Method】 In the spring maize growth period, field experiment were conducted with different irrigation quota (4 800, 4 200 and 3 600 m³·hm^-2) in the middle reach of the Shiyanghe Basin in 2014-2015. Soil water and salt space-time distribution of 0-100 cm soil layer, the interannual change of soil salt content and spring maize yield and its components were measured.【Result】 The more irrigation quota was , the more soil water content was in 0-60 cm soil layer. Increasing irrigation quota from 420 m³·hm^-2 to 480 m³·hm^-2, soil water content could be higher than 24.52% of average in 0-60 cm soil layer. In key water requirement period of crop, it could increase the deep soil moisture with 480 m³·hm^-2 of irrigation quota. While it had a serious soil moisture deficit with 360 m³·hm^-2 of irrigation quota. In non-irrigation period, soil salt accumulated in 0-40 cm layer and desalinated in 80-100 cm layer in vertical direction. In the irrigation period, desalt depth increased with increasing irrigation quota in vertical direction. Soil salt desalinated in 0-20 cm soil layer and accumulated in 40-100 cm soil layer, while total soil salt of 0-100 soil layer remained unchanged. In horizontal direction, soil salt migrated in both sides of drip irrigation belt, in addition, the effect of soil moisture movement migrated parts of soil salt which was been kept in arable layer to wetting front edge between the middle of the two drip irrigation belt in horizontal direction. Soil salt content in drip irrigation belt interval was higher than in drippers interval, irrigation quota had no significant effect on ear length, ear rows and rows grains of spring maize, while had significant effect on ear diameter, bald tip length and hundred-grain weight.【Conclusion】Under the condition of drip irrigation, water consumption of spring maize was mainly influenced by irrigation water. Water deficit in a certain extent could elevate water use efficiency (WUE), while yield of spring maize was reduced by 4.45%-20.99%. Irrigation schedule which had irrigation norms of 420 m³·hm^-2 and 10 irrigation frequencies was recommended as the most optimal result of saving water, preventing soil salt from moving up and increasing yield of spring maize.

Construction of SSR Genetic Linkage Map and Comparison on Pears

WANG Lei, WANG Long, XUE Hua-bai, LI Xiu-gen, LI Jiang

Scientia Agricultura Sinica. 2016, 49(12): 2353-2367. doi:10.3864/j.issn.0578-1752.2016.12.011

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【Objective】The genetic linkage map was constructed based on the Simple Sequence Repeat primers of pears and apples that have been published and developed from the transcriptome of pears by our team, which will be the foundation for the important trait mapping of QTL and marker-assisted selection in the future.【Method】A F₁ mapping population was built on a cross between a western pear (Red Clapp Favorite) as the female parent, and an oriental pear (Mansoo) as the male parent. The SSR primers which matched the "CP" mode in the JoinMap 4.0 software in parents and four individuals by PCR application were selected and then tested in the F₁ group. The separation of data was analyzed by JoinMap 4.0 software, and the parent genetic map was constructed respectively. The genetic map of the parents was integrated based on anchor points which were homology marks in the parents’ map on each linkages.【Result】909 pairs of SSR primers from different sources (526 pairs from pears and 283 pairs from apples that have been published, and 100 pairs were developed from the pear transcriptome by our team) was preliminarily screened through a PCR technique. The results showed that amplified fragments and polymorphism of SSR primers from apples were lower, but that the SSR primers from pears and developed through the transcriptome of pears were higher. The parents’ genetic linkage map was constructed based on the 207 pairs of SSR primers which conformed to the drawing. A female genetic map was constructed by 141 markers which was distributed on 17 linkage groups and spanned 757.34 cM in genome with an average distance of 5.37 cM between markers; A male genetic map was constructed by 153 markers which was distributed on 19 linkage groups and spanned 1 149.43 cM in genome with an average distance of 7.51 cM between markers.【Conclusion】The integrated genetic linkage maps of parents were constructed by different sources of SSR molecular markers, which contained 186 SSR markers and finally covered the genome 1 125.33 cM.

Analysis of Codon Usage Pattern of Paeonia lactiflora Genes Regulating Flower Color and Its Influence Factors

WU Yan-qing, ZHAO Da-qiu, TAO Jun

Scientia Agricultura Sinica. 2016, 49(12): 2368-2378. doi:10.3864/j.issn.0578-1752.2016.12.012

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【Objective】 The quality of Paeonia lactiflora flower color affects its ornamental value and the commercial value of ornamental plants. This study aims to understand the codon usage pattern of genes regulating flower color and probe into the main factors affecting the formation of codon bias, which has important biological significance for mRNA translation, design of transgenes, the prediction of expression level and functions of new genes, and studies of molecular biology and evolution, etc. 【Method】In a previous study, 6,345 differential genes were screened out by transcriptome sequencing of a flower color chimaera cultivar “Jinhui” with a consistent genetic background red outer-petal and yellow inner-petal, followed by a further filtering analysis according to the principle of CDS sequence characteristics and greater than 300 bp. We finally obtained 2,234 genes as our research object. Mobyle software was used to calculate different parameters for the codon usage, such as GC content, average GC content of the first and second positions (GC12), GC content of the third position (GC3s), effective number of codon (ENC), codon adaptation index (CAI), and relative synonymous codon usage (RSCU). Further analysis of a neutrality plot (GC12 vs. GC3), an ENC-GC3s plot, and a Parity Rule 2 (PR2) plot were performed. Additionally, we probed into the influence of mutational pressure and translational selection by a multivariate statistical analysis. Finally, we took 5% CAI value as high-expression and low-expression sample groups, then calculated the RSCU value, and analyzed the significant difference to determine the optimal codons by a chi-square test.【Result】The results showed that the GC content at the third position of codons was 46.37%. The GC content of most genes was mainly distributed between 30% and 55%. Neutrality analysis showed that there was a significant positive correlation (R²=0.202, P＜0.01) between GC3s and GC12 value. The ENC-plot showed most of the genes on or close to the expected curve, but also some points with low-ENC values were below it. The(ENCexp-ENCobs)/ENCexp ratio of most genes ranged from -0.05 to 0.05. The Parity Rule 2-plot showed that the frequency of T nucleotide at the third position was higher than A, and C was higher than G, suggesting that the use frequencies of four nucleotide were not balanced. Correspondence analysis showed that the first axis showed a 38.09% variation, while the other three axes showed 18.42%, 15.09%, and 14.59%, respectively, suggesting that the first axis was the main index evaluating the codon usage bias of Paeonia lactiflora genes regulating flower color. Mutation pressure and selection analysis showed there were significant negative correlations (R²=0.736, P＜0.01. R²=0.286, P＜0.01) between the first axis and GC3s, CAI value, respectively. Using the delta RSCU and significant chi-square test methods, we defined 21 codons as the major preference codons in the Paeonia lactiflora genes regulating flower color, of 18 codons ending with G or C, only CGU and GGU ending with U.【Conclusion】In conclusion, most of optimal codons ended with G or C. Meanwhile, the codon usage pattern of Paeonia lactiflora genes regulating flower color is formed under the effect of mutational pressure (R²=0.736) and translational selection (R²=0.286), but a mutational bias was the major influence on codon usage. This study not only preliminarily reveals the codon usage pattern of Paeonia lactiflora genes, but also provides a certain theoretical basis for further carrying out genetic improvement of Paeonia lactiflora flower color by codon reconstruction and analyzing the molecular evolution.

The Preparation Process for Isolation of a Highly Active Antioxidant Peptide Derived from Wheat Germ Albumin

CHEN Si-yuan, LIU Yong-xiang, CAO Xiao-zhou, ZHANG Yi-jing, CHEN Hai-juan, SHEN Xin-chun

Scientia Agricultura Sinica. 2016, 49(12): 2379-2388. doi:10.3864/j.issn.0578-1752.2016.12.013

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【Objective】 The aim of this study is to develop an effective preparation process for obtaining highly active antioxidant peptide derived from wheat germ albumin using two-step dual-enzymatic hydrolysis. 【Method】 A two-step dual-enzymatic hydrolysis and separation were employed to screen the antioxidant peptide (AOP) from wheat germ albumin. The in vitro AOP’s antioxidant activity was evaluated by DPPH radical scavenging activity and Fe²⁺ chelating ability assays. The hydrolysate derived from tryptic hydrolysis of wheat germ albumin was fractionated by ultrafiltration according to molecular weight; then the fractions with high antioxidant capacity were further isolated by gel filtration chromatography (Sephadex G-75). After that, the fraction with higher antioxidant activity was further hydrolyzed by an alcalase, and the hydrolysate was subjected to purification by Sephadex G-15 gel filtration chromatography and reversed-phase HPLC (RP-HPLC), respectively. The highest antioxidant fraction was achieved and the structure was identified by ESI-TOF MS/MS. 【Result】 Hydrolysate derived from tryptic hydrolysis of wheat germ albumin was fractionated by ultrafiltration according to molecular weight. Three fractions, P_a, P_b and P_c were obtained from the fraction with high antioxidant capacity. The fraction P_b showed the strongest antioxidant activity evaluated by DPPH radical scavenging activity and Fe²⁺ chelating ability assays and its extraction yield was 23.1%. The fraction P_b which had the highest antioxidant activity was further hydrolyzed by an alcalase, and the hydrolysate was subjected to gel filtration chromatography (Sephadex G-15) and two more groups, P_d and P_e were obtained. The extraction yield of fraction P_d which had higher antioxidant capacity was 52.1%. After further purification by RP-HPLC, five fractions, P₁, P₂, P₃, P₄ and P₅ were separated from the fraction P_d, and the fraction P₃ exhibited the highest antioxidant activity with the in vitro EC₅₀ values of DPPH radical scavenging activity at 1 mg/ml and Fe²⁺ chelating ability at 0.6 mg/ml, respectively. The extraction yield of the fraction P₃ was 7.3% in RP-HPLC step. Finally, employing the electrospray ionization-time-of-flight mass spectrometer (ESI-TOF MS/MS), the structure of the fraction P₃ was identified. Its amino acids sequence, molecular weight and purity were AREGETVVPG, 1013.51Da and 85%, respectively. Consistently, the antioxidant capacity of the synthetic peptide with the same sequences ( purity ≥95%) showed the similar activity to the fraction P₃ (P>0.05). 【Conclusion】 The preparation process of a two-step dual-enzymatic hydrolysis in combination with ultrafiltration, gel filtration chromatography, and RP-HPLC was effective in separation, screen and purification of a highly active antioxidant peptide.

Research on the Expression and Localization of CART in Bovine Granulosa Cells at Different Developmental Stages

LI Peng-fei, BI Xi-lin, WANG Kai, JING Jiong-jie, Lü Li-hua

Scientia Agricultura Sinica. 2016, 49(12): 2389-2396. doi:10.3864/j.issn.0578-1752.2016.12.014

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【Objective】The objective of this paper is to investigate the effect of CART (cocaine-and amphetamine- regulated transcript) expression in granulosa cells on bovine follicular development at different stages.【Method】 Two RNA libraries from ODF1 (the largest onset of deviation follicle) and ODF2 (the second largest onset of deviation follicle) granulosa cells were prepared and sequenced using Illumina HiSeq 2000 platform. Cows were slaughtered, and both ovaries were collected, then the largest and the second largest follicles were screened through the morphology of corpus luteum in the first follicular wave. Follicular fluid was extracted for the concentration of E2 and P measurement using competitive ELISA method to determine DF (dominant follicles) and SF (subordinate follicles). Granulesa cells were isolated from DF and SF, and total RNA were extracted, respectively. Using RPLP0 as a reference gene, primers were designed. qRT-PCR was performed to detect the expression of CART in DF and SF, GraphPad Prism 5.0 was used for construction and significant analysis. Rabbit anti-CART antibody was used to detect the CART expression in cow follicles.【Result】Deep sequencing results showed that CART amounts in ODF1 were 38.2 times more than that in ODF2. Ovaries of 3 cows were screened out which were in the first follicular wave according to the observation of the morphology of corpus luteum, and the DF and SF were obtained. Real-time PCR results showed that CART expressed in DF was extremely significantly higher than that in SF, up to 2310-fold, which is consistent with the deep sequencing results. Immunohistochemistry results showed that CART expressed both in DF and SF, and the amounts in DF was significantly greater than that in SF, at the same time, CART was mainly expressed in granulosa cell layer.【Conclusion】During the process of follicular development in cattle, other inhibitors have suppressed most of the follicles, and prevented them to develop into DF; when the deviation appears, CART inhibited DF to further develop into ovulatory follicle, so it is considered that CART possibly by promoting granulosa cells apoptosis, and suppresses the E₂ secretion in granulosa cell layer, eventually leads to DF atresia.

Study of Location and Distribution of Classical Swine Fever Virus RNA in PK15 Cells by Visualization in Situ Hybridization Technology

ZHANG Yu-jie, ZHAO Yan, XU Lu, ZHANG Qian-yi, CHEN Kai, SUN Yong-fang, ZOU Xing-qi, ZHU Yuan-yuan, ZHAO Qi-zu, NING Yi-bao, WANG Qin

Scientia Agricultura Sinica. 2016, 49(12): 2397-2407. doi:10.3864/j.issn.0578-1752.2016.12.015

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【Objective】In order to make researches on the RNA location and distribution of Classical Swine Fever Virus （CSFV）in infected PK15 cells, a rapid, accurate and sensitive in situ hybridization (ISH) technology was established. 【Method】After comparison with the complete sequences of CSFV, BVDV and BDV to avoid the homology regions, a set of specific probes of CSFV RNA andβ-actin were designed and synthesized. A reference strain CSFV (HeBHH1/95) was used to optimize the ISH technology through comparison several parameters such as fluorescence intensity, repeatability, protease K concentration and formalin fixation time. After these parameters were determined, an ISH technology was established. Fluorescent antibody test (FAT) was used to compare the sensitivity with the ISH technology. All the sub genotypes of CSFV (sub genotype 1.1, 2.1,2.2 and 2.3) which are present in China and other normal pig infectious virus (BVDV, PPV, PRV，PCV-2) were used to detect specificity of this ISH technology. A high virulent strain of CSFV (SM) was inoculated in PK15 cells. Infected cells were sampled at 0.5 hours post inoculation (hpi)、1, 3, 6, 8, 10, 14, 18, 24, 36, 48, 72, and 96hpi. Then FAT was performed in parallel to detect the expression and location of CSFV E2 protein. 【Result】CSFV RNA were detected in infected PK15 cells by using the ISH technology. The optimal concentration of protease K was 1:1 000 and the optimal time of formalin fixation was 30 minutes. The minimum of detection is 10-8/200μL which is 3.5 orders of magnitude higher than FAT. The specific tests showed that the ISH technology could react with sub genotypes 1.1, 2.1,2.2 and 2.3 of CSFV in China and has no cross reaction with BVDV, PPV, PRV，PCV-2 viruses. CSFV RNA ISH test results showed that CSFV RNA were firstly detected in nucleus at 0.5 hpi and gathered in nucleus from 0.5hpi to 6hpi; at 10hpi, there were more CSFV RNA gathered in cytolymph than before and less CSFV RNA were detected than before in nucleus；at 24hpi, CSFV RNA mainly gathered in cytolymph around nucleus; at 36hpi, more and more CSFV RNA gathered in cytolymph and got maximum at 72hpi; at 96hpi, the amount of CSFV RNA declined. The results of FAT showed that little E2 protein was detected in cytolymph at 8hpi. From 10hpi to 24 hpi, only few cells were detected positive. After 36hpi, the expression of E2 protein increased gradually and got to the maximum at 72hpi. But after 96hpi, the amount of E2 protein declined. The trend of E2 protein expression was in accord with CSFV RNA. 【Conclusion】 A visualization in situ hybridization technology was firstly established to detect CSFV RNA. The location and distribution of CSFV RNA was studied by using this technology. The results proved that CSFV RNA enters cell early than 0.5hpi and CSFV RNA has ever existed in nucleus.

Cytological Researches on the Anther Development of a Thermo-Sensitive Genic Male Sterile Line TE5A in Brassica napus

LI Ke-qi, ZENG Xin-hua, YUAN Rong, YAN Xiao-hong, WU Gang

Scientia Agricultura Sinica. 2016, 49(12): 2408-2417. doi:10.3864/j.issn.0578-1752.2016.12.016

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【Objective】The aim of this paper is to detect the vital abortion periods and cytological characteristics of the anther development in a thermo-sensitive genic male sterile line TE5A in Brassica napus. The results will provide a theoretical basis for further study on the mechanism of male sterility.【Method】The sterile line TE5A was used as test material and was conducted with two different treatment temperatures (16℃ and 22℃). Then the fertility and characteristics of the floral organs of the plants were observed, respectively. The differences of anther development between the sterile and the normal plants of the sterile line TE5A were observed and compared by using toluidine blue, aniline blue and sudan black B to stain the semi-thin sectionings. Further observations and comparisons of cell ultrastructure of the anther development were carried out by transmission electron microscopy.【Result】The flowers of the plants were observed to be sterile, after seven days when the fertile rapeseed plants were moved to the light incubator of 22℃ from the one of 16℃. After the sterile rapeseed plants were moved to the light incubator of 16℃ from the one of 22℃, it was observed that the sterile flowers were opened first and the fertile flowers were opened after. Compared with the fertile flowers, the size and characteristic of petals of sterile plants in the sterile environment displayed no obvious difference. But the filaments of the sterile plants were significantly shorter than the fertile's and the sterile anthers withered without pollen grains in them. The pollen abortion of the sterile line TE5A in the sterile environment occurred at the pollen mother cell meiosis stage. The pollen mother cells could not pass the meiosis, with no dyads and tetrads formed in TE5A, insteading of some "pseudo microspores". The callose was normal deposited around the pollen mother cells, however, the callose hadn't been degraded until the mature pollen stage. The tapetum cells which could secret liposomes were observed to be normal in the fertile line. The pollen exines of the "pseudo microspores" were abnormal. The special bacula and tectum structure of the pollen exine couldn't formed, so the pollen exine couldn't combine with sporopollenins, no liposomes and other substances combined. With the development of the anthers, the "pseudo microspores" were decayed gradually and only empty shells remained at last.【Conclusion】Compared with the fertile flowers, the size and characteristic of petals of sterile plants in the sterile line TE5A displayed no obvious difference. But the filaments of the sterile plants were significantly shorter and the sterile anthers withered without pollen grains in them. The vital abortion of the sterile line TE5A occurred at the pollen mother cell meiosis stage. The pollen mother cells couldn’t pass the meiosis, and neither dyads nor tetrads were formed. The abnormal degradation of the callose also play an important role in the anther development. The cytological characteristics of the thermo-sensitive genic male sterile line TE5A in B. napus were different from the previous materials. So these indicated that TE5A is a novel thermo-sensitive genic male sterile line.

Cloning and Expression Analysis of Two DREB1/CBF Genes in Suaeda salsa L.

SUN Xiao-bo, SU Jia-le, JIA Xin-ping, LIANG Li-jian, XIAO Zheng, DENG Yan-ming

Scientia Agricultura Sinica. 2016, 49(12): 2418-2429. doi:10.3864/j.issn.0578-1752.2016.12.017

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【Objective】To provide basic information for understanding the resistant mechanism under abiotic stresses, two DREB1/CBF genes were cloned from Suaeda salsa L. At the same time, the sequence characteristics, subcellular localization and transcriptional activities of the two predicted DREB1/CBF proteins and their expression alteration in response to abiotic stress were analyzed. 【Method】The fragments of two DREB1/CBF genes were obtained by using the technique of homologous cloning. The full-length of cDNA sequences of the two DREB1/CBF genes were isolated by the method of rapid-amplification of cDNA ends (RACE), and they were named as SsCBF1 and SsCBF2, respectively. The structures and functions of the two proteins encoded by SsCBFswere predicted by the bioinformatics software. In order to detect the subcellular localization of the two proteins, the coding sequences of the two SsCBF genes were fused, respectively, downstream to the GFP sequence to obtain two expression vectors and they were separately transferred into onion epidermal cells by the biolistic method. The binding specificity of SsCBF1 and SsCBF2 to DRE/CRT cis-acting element and their transcriptional activities were investigated by using a yeast one-hybrid system. The expression of the two SsCBF genesin response to low temperature, NaCl，PEG and ABA were assessed by the real-time quantitative PCR.【Result】 SsCBF1 encoded a peptide of 225 amino acid residues with a predicted molecular mass of 25.4 kD and a pI of 4.84. SsCBF2 encoded a predicted protein of 260 amino acid residues with a predicted molecular mass of 28.6 kD and a calculated pI of 5.05. SsCBF1and SsCBF2both contained a typical AP2/ ERF domain andshared 53.5% and 45.4% identity at the levels of coding nucleotides and amino acids. However, the AP2/ ERF domains of the two SsCBF genes were highly similar and shared 76.2% and 87.3% identity at the levels of nucleotides and amino acids, respectively. SsCBF1 and SsCBF2 were classified into A-1 subgroup of the DREB subfamily and both localized to the nucleus. The two proteins were able to specifically bind to the DRE/CRT sequence and activate the expression of the down-stream HIS reporter gene in yeast. Low temperature, drought, high salt and ABA could induce the expression of SsCBF1. However, the expression of SsCBF2 could only be induced significantly by low temperature, not by drought, high salt and ABA. 【Conclusion】The SsCBF1 and SsCBF2 are two stress-responsive transcription factors of S. salsa. In S. salsa, SsCBF1 is involved in the stress responses of cold, high-salt and drought through ABA-dependent pathways and SsCBF2 is responsive to cold stress through ABA-independent pathway.

Effect of Cage Type on Slaughter Performance and Meat Quality Change During Postmortem Aging of Meat Rabbit

ZHOU Qin-fei, HE Zhi-fei, LI Hong-jun

Scientia Agricultura Sinica. 2016, 49(12): 2430-2436. doi:10.3864/j.issn.0578-1752.2016.12.018

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【Objective】 Aimed to provide reference for the selecting of meat rabbit breeding pens construction and meat consumption, the carcass traits and meat quality changing during postmortem aging of meat rabbit that reared in monolayer large cage and three layer cage were detected in the study. 【Method】 10 California rabbits (half male and half female) , reared in the same building, 70 days, health and the weight close to the average population, in each monolayer large cage and three layer cage were selected and slaughtered as the rabbit slaughtering methods, then the slaughter performance and the pH value, color, shear force, cooking loss and water loss of muscle during postmortem aging were determined. 【Result】 The influence of two kinds of cages on the slaughter weight was not significant. Compared to the monolayer large cage, rabbits reared in the three layer cage had a significant decrease in fore part and increase in hind part ratio of eviscerated carcass (P<0.05), a significant decrease in abdominal fat ratio (P<0.01), and a significant decrease in the eviscerated and semi-eviscerated (P<0.05). Liver weight and kidney weight were higher in the three layer cage reared rabbits. Femur weight, tibia weight and muscle ratio of hind leg were higher in monolayer large cage reared rabbits. The time of maximum muscle L * value during postmortem aging of the three layer cage reared rabbits was at the 2nd day, and the monolayer large cage reared rabbits was at the 3rd day. The muscle L* value of the three layer cage reared rabbits at the 6th day was significantly higher than the monolayer large cage (P<0.05), and very significantly higher at the 7th day (P<0.01), but it was lower at 3 days before. The muscle a* value during postmortem aging of monolayer large cage reared rabbits was significantly higher than the three layer cage at the 2nd day, 3rd day and 5th day (P<0.05), and it was very significantly higher at the 4th day (P<0.01). The muscle b* value during postmortem aging of the three layer cage reared rabbits was higher than the monolayer large cage, and it was very significantly higher at the 1^st day (P<0.01). The muscle pH value during postmortem aging of the three layer cage reared rabbits was higher than the monolayer large cage at the 3 days before, and lower after 3 days. The monolayer large cage reared rabbits had a significantly higher muscle shear force at the 1st and 3rd day during postmortem aging compared to the three layer cage (P<0.05), and very significant at the 2nd day (P<0.01), but lower after the 4th day. The three layer cage reared rabbits had a higher cooking loss compared to monolayer large cage reared rabbits during the postmortem aging. And the water loss index changing during the postmortem aging was in agreement with cooking loss. 【Conclusion】 Monolayer large cages can change the distribution of muscle in the rabbit carcass , and have a positive effect on meat quality compared to the three layer cage.

Analysis of Effect Factors on SCC and Milk Production Traits of Xinjiang Brown Cattle

TIAN Yue-zhen, FENG Wen, WANG Ya-chun, HUANG Xi-xia,YU Ying

Scientia Agricultura Sinica. 2016, 49(12): 2437-2448. doi:10.3864/j.issn.0578-1752.2016.12.019

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【Objective】The objective of this study was to analyze the non-genetic and co-variable effect factors of somatic cell counts (SCC) and milk production traits of Xinjiang brown cattle, and the results of the study will supply a scientific basis for guidance of improving production performance of Xinjiang brown cattle.【Method】SAS9.2 was used to analyze the effect of herd, parity and lactation month on SCC and milk production traits as well as their regression with each other in 262 Xinjiang brown cattle in two farms. And then, with Excel 2 013 software, 1 510 dairy herd improvement (DHI) records and the SCC deviation values between adjacent two months were used to draw the figures about the distribution of SCC and their regression with milk production traits of Xinjiang brown cattle in two farms, respectively. 【Result】 The herd and parity had an extremely significant effect on somatic cell counts, all of the milk production traits as well (P＜0.01). Lactation month had an extremely significant effect on milk yield and SCC (P＜0.01), had a significant effect on sugar percentages (P＜0.05) while had no effect on other milk components (P＞0.05). Milk yield, fat percentages, protein percentages and sugar percentages decreased following the increase of SCC, while the total dry matters content increased. The interaction effect among herd, parity and lactation month had no effect on SCC（P＞0.05）. The interaction of herd and parity had an extremely significant effect on milk yield and all of the milk components (P＜0.01). The interaction of herd and lactation month had an extremely significant effect on fat percentage and total dry matters content (P＜0.01), had a significant effect on sugar percentages (P＜0.05) while had no effect on other milk components (P＞0.05). The interaction of parity and lactation month had no effect on any trait. According to the figures of lactation curve which appeared regulations on variations of SCC deviation value and milk production traits. As the higher SCC deviation value, the lower mean of milk yield and sugar percentage. SCC difference ranking method can be used to evaluate the production of dairy cow effectively and determine the trend of mastitis. In comparison of the two farms, it was found that the second farm was better than the first farm as its higher milk production and milk components, lower SCC and the better management level. 【Conclusion】 In order to increase the quality and yield of milk, effective measures should be taken, such as improving milking management, hygiene and standard operation of milking facilities. In addition, SCC deviation value between two months could response the healthy condition of dairy cattle effectively, and it will avoid infecting mastitis by controlling the somatic cell counts in cows. Production management level and DHI monitoring production situation should be strengthened in the first farm to control the SCC and avoid recessive mastitis in Xinjiang brown cattle.

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