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2016 Vol. 15 No. 3 Previous Issue    Next Issue

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Crop Genetics · Breeding · Germplasm Resources
Physiology·Biochemistry·Cultivation·Tillage
Plant Protection
Animal Science · Veterinary Science
Soil & Fertilization﹒Irrigation﹒Plant Nutrition﹒ Agro-Ecology & Environment
Food Science
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Crop Genetics · Breeding · Germplasm Resources

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A genetic diversity assessment of starch quality traits in rice landraces from the Taihu basin, China AO Yan, XU Yong, CUI Xiao-fen, WANG An, TENG Fei, SHEN Li-qun, LIU Qiao-quan 2016, 15(3): 493-501.  DOI: 10.1016/S2095-3119(15)61050-4 Abstract ( )   PDF in ScienceDirect   There are nearly 1 000 rice landrace varieties in the Taihu basin, China. To assess the genetic diversity of the rice, 24 intragenic molecular markers (representing 17 starch synthesis-related genes) were investigated in 115 Taihu basin rice landraces and 87 improved cultivars simultaneously. The results show that the average genetic diversity and polymorphism information content values of the landraces were higher than those of improved cultivars. In total, 41 and 39 allele combinations (of the 17 genes) were derived from the landraces and improved cultivars, respectively; only two identical allele combinations were found between the two rice variety sources. Cluster analysis, based on the molecular markers, revealed that the rice varieties could be subdivided into five groups and, within these, the japonica improved rice and japonica landrace rice varieties were in two separate groups. According to the quality reference criteria to classify the rice into grades, some of the landraces were found to perform well, in terms of starch quality. For example, according to NY/T595-2002 criteria from the Ministry of Agriculture of China, 25 and 33 landraces reached grade 1, in terms of their apparent amylose content and gel consistency. The varieties that had outstanding quality could be used as breeding materials for rice quality breeding programs in the future. Our study is useful for future applications, such as genetic diversity studies, the protection of rice variety and improvment of rice quality in breeding programs.

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Characterization of Ppd-D1 alleles on the developmental traits and rhythmic expression of photoperiod genes in common wheat ZHAO Yong-ying, WANG Xiang, WEI Li, WANG Jing-xuan, YIN Jun 2016, 15(3): 502-511.  DOI: 10.1016/S2095-3119(15)61129-7 Abstract ( )   PDF in ScienceDirect   Photoperiodic response is an important characteristic that plays an important role in plant adaptability for various environments. Wheat cultivars grow widely and have high yield potential for the strong photoperiod adaptibility. To assess the photoperiodic response of different genotypes in wheat cultivars, the photoperiodic effects of the Ppd-D1 alleles and the expressions of the related TaGI, TaCO and TaFT genes in Liaochun 10 and Ningchun 36 were investigated under the short-day (6 h light, SD), moderate-day (12 h light, MD) and long-day (24 h light, LD) conditions. Amplicon length comparison indicated that the promoter of Ppd-D1 in Ningchun 36 is intact, while Liaochun 10 presented the partial sequence deletion of Ppd-D1 promoter. The durations of all developmental stages of the two cultivars were reduced by subjection to an extended photoperiod, except for the stamen and pistil differentiation stage in the Liaochun 10 cultivar. The expression levels of the Ppd-D1 alleles and the TaGI, TaCO and TaFT genes associated with the photoperiod pathway were examined over a 24-h period under SD and MD conditions. The relationships of different photoperiodic responses of the two cultivars and the expression of photoperiod pathway genes were analyzed accordingly. The photoperiod insensitive (PI) genotype plants flower early under SD; meanwhile, the abnormal expression of the Ppd-D1a allele is accompanied with an increase in TaFT1 expression and the TaCO expression variation. The results would facilitate molecular breeding in wheat.

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Development and identification of Verticillium wilt-resistant upland cotton accessions by pyramiding QTL related to resistance GUO Xiu-hua, CAI Cai-ping, YUAN Dong-dong, ZHANG Ren-shan, XI Jing-long, GUO Wang-zhen 2016, 15(3): 512-520.  DOI: 10.1016/S2095-3119(15)61083-8 Abstract ( )   PDF in ScienceDirect   Cotton Verticillium wilt is a serious soil-borne disease that leads to significant losses in fiber yield and quality worldwide. Currently, the most effective way to increase Verticillium wilt resistance is to develop new resistant cotton varieties. Lines 5026 and 60182 are two Verticillium wilt-resistant upland cotton accessions. We previously identified a total of 25 quantitative trait loci (QTLs) related to Verticillium wilt resistance from 5026 and 60182 by assembling segregating populations from hybridization with susceptible parents. In the current study, using 13 microsatellite markers flanking QTLs related to Verticillium wilt resistance, we developed 155 cotton inbred lines by pyramiding different QTLs related to Verticillium wilt resistance from a filial generation produced by crossing 5026 and 60182. By examining each allele’s effect and performing multiple comparison analysis, we detected four elite QTLs/alleles (q-5/NAU905-2, q-6/NAU2754-2, q-8/NAU3053-1 and q-13/NAU6598-1) significant for Verticillium wilt resistance, pyramiding these elite alleles increased the disease resistance of inbred lines. Furthermore, we selected 34 elite inbred lines, including five lines simultaneously performing elite fiber quality, high yield and resistance to V. dahliae, 14 lines with elite fiber quality and disease resistance, three lines with high yield and disease resistance, and 12 lines with resistance to V. dahliae. No correlation between Verticillium wilt resistance and fiber quality traits/yield and its components was detected in the 155 developed inbred lines. Our results provide candidate markers for disease resistance for use in marker-assisted breeding (MAS), as well as elite germplasms for improving important agronomic traits via modern cotton breeding.

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Hybridization between salt resistant and salt susceptible genotypes of mungbean (Vigna radiata L. Wilczek) and purity testing of the hybrids using SSRs markers Sehrawat Nirmala, Yadav Mukesh, Bhat Kangila Venkataraman, Sairam Raj Kumar, Jaiwal Pawan Kumar 2016, 15(3): 521-527.  DOI: 10.1016/S2095-3119(15)61161-3 Abstract ( )   PDF in ScienceDirect   Six cultivated and two wild genotypes of mungbean (Vigna radiata L. Wilczek) possessing variation for phenotypic and genotypic response for salt tolerance were hybridized. Hybridization results showed successful pod set and significant variations in cross-compatibility of investigated genotypes. Genotypes PLM 380 and PLM 562 showed promising combining ability with all genotypes. Results revealed significant crossing compatibility between V. radiata and V. sublobata. The cross ability ranged from 1.99 to 5.12% (average 3.08%). Molecular analysis confirmed the hybrids purity. All F1 seeds were bold, green/shiny green and germinated between 3 to 5 days. Hybrid plants were normal, fertile and healthier over their respective progenitors. Uniform flowering and maturity of the hybrids showed absence of any defect or alteration in plant habit and life cycle of the hybrids. The hybrid plants showed increase in yield characteristics as no. of pods, pod length, 100 seeds weight, and yield per plant. Number of pods containing F2 seeds ranged from 5–8 per cluster. The F2 seeds were collected and stored for further research. Present study suggests that salt resistant wild relatives or cultivars of mungbean can be explored by breeding as a source of useful traits/genes providing salt tolerance. This may help in development of best mungbean variety for saline prone areas. Micro-satellite markers linked to the trait/genes may assist breeding by early selection of the genotypes compared to the phenotypic screening.

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Isolation and characterization of an ERF-B3 gene associated with flower abnormalities in non-heading Chinese cabbage XU Yu-chao, HOU Xi-lin, XU Wei-wei, SHEN Lu-lu, Lü Shan-wu, ZHANG Shi-lin, HU Chun-mei 2016, 15(3): 528-536.  DOI: 10.1016/S2095-3119(15)61203-5 Abstract ( )   PDF in ScienceDirect   BrcERF-B3 gene, a member of ethylene-responsive factor family, was screened from a mutant plant in non-heading Chinese cabbage (Brassica rapa ssp. chinensis) by cDNA-AFLP technology. We got full length cDNA of two BrcERF-B3 genes by homology-based cloning from two materials and found that their nucleotide sequences were the same by sequencing. The BrcERF-B3 protein, belonging to the B3 subgroup of the ERF subfamily, shared a close relationship with B. rapa. RT-PCR result showed that BrcERF-B3 expressed only in mutant stamen rather than maintainer stamen. qRT-PCR results indicated that BrcERF-B3 expressed highly during reproductive growth development and in the early of mutant buds, suggesting BrcERF-B3 might be involved in the formation of abnormal flower in mutant. What’s more, the expression of BrcERF-B3 was more significant to ABA, MeJA and cold stresses in mutant than in maintainer and was down-regulated in NaCl treatment in two lines, implying BrcERF-B3 might be different roles in biotic and abiotic stresses.

Physiology·Biochemistry·Cultivation·Tillage

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Charactering protein fraction concentrations as influenced by nitrogen application in low-glutelin rice cultivars LI Gang-hua, CHEN Yi-lu, DING Yan-feng, GENG Chun-miao, LI Quan, LIU Zheng-hui, WANG Shao-hua, TANG She 2016, 15(3): 537-544.  DOI: 10.1016/S2095-3119(15)61182-0 Abstract ( )   PDF in ScienceDirect   To optimize both grain yield and quality of low-glutelin rice cultivars under N-fertilizer strategies, two-year field experiments involving three low-glutelin rice cultivars (W1240, W1721, W025) and an ordinary rice cultivar (H9405) with five N treatments were carried out to determine the effects of N application rate and genotype on protein fractions contents and Glutelin/Prolamin ratio (Glu/Pro). The difference of protein fraction concentrations affected by N application rate existed in genotypes. Ordinary rice cultivar had a larger increase in glutlein concentration affected by N application rate than low-glutelin rice cultivars did. Glutelin in H9405 had a increase of 30.6 and 41.0% under the N4 treatment (360 kg N ha–1) when compared with N0 treatment (no fertilizer N) in 2010 and 2011 respectively, while all the low-glutelin rice cultivars showed relatively smaller increases for two years. Variance analysis showed no significant effect of N application rate on glutelin in W1240 and W025 while the effects on albumin, globulin and prolamin were significant in low-glutelin rice. What’s more, N treatment had no significant influence on Glu/Pro ratios in low-glutelin rice cultivars while a significant increase in Glu/Pro ratio was observed in ordinary rice cultivar. So low-gultelin rice cultivars showed a different pattern from ordinary rice cultivars when influenced by N application rate.

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Photosynthetic characteristics and nitrogen distribution of largespike wheat in Northwest China WANG Li-fang, CHEN Juan, SHANGGUAN Zhou-ping 2016, 15(3): 545-552.  DOI: 10.1016/S2095-3119(15)61151-0 Abstract ( )   PDF in ScienceDirect   The leaf photosynthesis and nitrogen (N) translocation in three large-spike lines and control cultivar (Xi’nong 979) of winter wheat (Triticum aestivum L.) were studied in 2010–2011 and 2011–2012. The objectives of this study were to investigate the differences in the physiological characteristics of large-spike lines and control cultivar and identify the limiting factors that play a role in improving the yield of breeding materials. The average yield, grain number per spike, kernel weight per spike, and 1 000-kernel weight of the large-spike lines were 16.0, 26.8, 42.6, and 15.4%, respectively, significantly higher than those of control. The average photosynthetic rates (Pn) were not significant between the large-spike lines and control cultivar during the active growth period. The average PSII maximum energy conversion efficiency (Fv/Fm), PSII actual quantum efficiency (ФPSII), photochemical quenching coefficient (qP), PSII reaction center activity (Fv´/Fm´) and water-use efficiency (WUE) of the large-spike lines were 1.0, 5.1, 3.6, 0.8, and 43.4%, respectively, higher than those of the control during the active growth stages. The N distribution proportions in different tissues were ranked in the order of grains>culms+sheathes>rachis+ glumes>flag leaves>penultimate leaves>remain leaves. This study suggested that utilization of the large-spike wheat might be a promising approach to obtain higher grain yield in Northwest China.

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Winter cover crops alter methanotrophs community structure in a double-rice paddy soil LIU Jing-na, ZHU Bo, YI Li-xia, DAI Hong-cui, XU He-shui, ZHANG Kai, HU Yue-gao, ZENG Zhao-hai 2016, 15(3): 553-565.  DOI: 10.1016/S2095-3119(15)61206-0 Abstract ( )   PDF in ScienceDirect   Methanotrophs play a vital role in the mitigation of methane emission from soils. However, the influences of cover crops incorporation on paddy soil methanotrophic community structure have not been fully understood. In this study, the impacts of two winter cover crops (Chinese milk vetch (Astragalus sinicus L.) and ryegrass (Lolium multiflorum Lam.), representing leguminous and non-leguminous cover crops, respectively) on community structure and abundance of methanotrophs were evaluated by using PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and real-time PCR technology in a double-rice cropping system from South China. Four treatments were established in a completely randomized block design: 1) double-rice cropping without nitrogen fertilizer application, CK; 2) double-rice cropping with chemical nitrogen fertilizer application (200 kg ha–1 urea for entire double-rice season), CF; 3) Chinese milk vetch cropping followed by double-rice cultivation with Chinese milk vetch incorporation, MV; 4) ryegrass cropping followed by double-rice cultivation with ryegrass incorporation, RG. Results showed that cultivating Chinese milk vetch and ryegrass in fallow season decreased soil bulk density and increased rice yield in different extents by comparison with CK. Additionally, methanotrophic bacterial abundance and community structure changed significantly with rice growth. Methanotrophic bacterial pmoA gene copies in four treatments were higher during late-rice season (3.18×107 to 10.28×107 copies g–1 dry soil) by comparison with early-rice season (2.1×107 to 9.62×107 copies g–1 dry soil). Type I methanotrophs absolutely predominated during early-rice season. However, the advantage of type I methanotrophs kept narrowing during entire double-rice season and both types I and II methanotrophs dominated at later stage of late-rice.

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Effects of selenium and sulfur on antioxidants and physiological parameters of garlic plants during senescence CHENG Bo, LIAN Hai-feng, LIU Ying-ying, YU Xin-hui, SUN Ya-li, SUN Xiu-dong, SHI Qing-hua, LIU Shi-qi 2016, 15(3): 566-572.  DOI: 10.1016/S2095-3119(15)61201-1 Abstract ( )   PDF in ScienceDirect   A hydroponic study was conducted to determine the effects of selenium (Se: 0, 3, 6 μmol L−1) on senescence-related oxidative stress in garlic plants grown under two sulfur (S) levels. We evaluated the yields of plants harvested at 160 and 200 days after sowing. Plants grown under a low Se dose (0.3 μmol L−1) at low S level showed higher yields (12.0% increase in fresh weight yield, 13.7% increase in dry weight yield) than the controls, despite a decrease in chlorophyll concentration. Compared with control plants, the Se-treated plants showed lower levels of lipid peroxidation. The Se-treated plants also showed higher activities of glutathione peroxidase and catalase, but lower superoxide dismutase activities. Changes in Fv/Fm values and proline contents were affected more strongly by S than by Se. On the basis of our results, we can conclude that Se plays a key role in the antioxidant systems in garlic seedlings. It delays senescence by alleviating the peroxide stress, but it can be toxic at high levels. A high S level may increase tolerance to high Se concentrations through reducing Se accumulation in plants.

Plant Protection

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Species-specific PCR-based assays for identification and detection of Botryosphaeriaceae species causing stem blight on blueberry in China XU Cheng-nan, ZHANG Hong-jun, CHI Fu-mei, JI Zhi-rui, DONG Qing-long, CAO Ke-qiang, ZHOU Zong-shan 2016, 15(3): 573-579.  DOI: 10.1016/S2095-3119(15)61177-7 Abstract ( )   PDF in ScienceDirect   Botryosphaeriaceae species are important causal agents of blueberry stem blight worldwide. Blueberry stem blight has become an important disease, potentially affecting the quality and production of blueberries in China. It is difficult and time-consuming to identify at the species level using morphological methods. The aim of this study was to develop polymerase chain reaction (PCR) assays for the diagnosis and early detection of latent infections of blueberry stems by Botryosphaeria spp. Species-specific primers, based on the ribosomal DNA internal transcribed spacer region and β-tubulin gene, were designed and selected for use in PCR assays. Three primer pairs, Lt347-F/R for Lasiodiplodia theobromae, Np304-F/R for Neofusicoccum parvum and FaF/Bt2b for Botryosphaeria dothidea, successfully amplified specific PCR fragments of different sizes on pure cultures or from blueberry stems inoculated and naturally infected blueberry plants with three pathogens, respectively. These primers did not amplify any PCR fragments from other blueberry stem disease-associated pathogens, such as Phomopsis spp. and Pestalotiopsis spp. This PCR protocol could detect as low as 100 pg to 1 ng of purified fungal DNA. This PCR-based protocol could be used for the diagnosis and detection of these pathogens from pure cultures or from infected blueberry plants.

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Ligand-binding properties of three odorant-binding proteins of the diamondback moth Plutella xylostella ZHU Jiao, Paolo Pelosi, LIU Yang, LIN Ke-jian, YUAN Hai-bin, WANG Gui-rong 2016, 15(3): 580-590.  DOI: 10.1016/S2095-3119(15)61067-X Abstract ( )   PDF in ScienceDirect   Strategies for insect population control are currently targeting chemical communication at the molecular level. The diamondback moth Plutella xylostella represents one of the most serious pests in agriculture, however detailed information on the proteins mediating olfaction in this species is still poor. This species is endowed with a repertoire of a large number of olfactory receptors and odorant binding proteins (OBPs). As a contribution to map the specificities of these chemical sensors in the moth and eventually unravel the complexity of chemodetection, we have measured the affinities of three selected OBPs to a series of potential odorants. Three proteins are highly divergent in their amino acid sequences and show markedly different expression profiles. In fact, PxylOBP3 is exclusively expressed in the antennae of both sexes, PxylOBP9 is male specific and present only in antennae and reproductive organs, while PxylOBP19, an unusual OBP with nine cysteines, is ubiquitously present in all the organs examined. Such expression pattern suggests that the last two proteins may be involved in non-chemosensory functions. Despite such differences, the three OBPs exhibit similar binding spectra, together with high selectivity. Among the 26 natural compounds tested, only two proved to be good ligands, retinol and coniferyl aldehyde. This second compound is particularly interesting being part of the chemical pathway leading to regeneration of lignin, one of the defense strategies of the plant against insect attack, and might find applications as a repellent for P. xylostella and other pests.

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Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens ZHANG Jiao, XING Yan-ru, HOU Bo-feng, YUAN Zhu-ting, LI Yao, JIE Wen-cai, SUN Yang, LI Fei 2016, 15(3): 591-599.  DOI: 10.1016/S2095-3119(15)61180-7 Abstract ( )   PDF in ScienceDirect   Methylation of the N6 position of adenine, termed N6-methyladenine, protects DNA from restriction endonucleases via the host-specific restriction-modification system. N6-methyladenine was discovered and has been well studied in bacteria. N6-adenine-specific DNA methyltransferase (N6AMT) is the main enzyme catalyzing the methylation of the adenine base and knowledge of this enzyme was mainly derived from work in prokaryotic models. However, large-scale gene discovery at the genome level in many model organisms indicated that the N6AMT gene also exists in eukaryotes, such as humans, mice, fruit flies and plants. Here, we cloned a N6AMT gene from Nilaparvata lugens (Nlu-N6AMT) and amplified its fulllength transcript. Then, we carried out a systematic investigation of N6AMT in 33 publically available insect genomes, indicating that all studied insects had N6AMT. Genomic structure analysis showed that insect N6AMT has short introns compared with the mammalian homologs. Domain and phylogenetic analysis indicated that insect N6AMT had a conserved N6-adenineMlase domain that is specific to catalyze the adenine methylation. Nlu-N6AMT was highly expressed in the adult female. We knocked down Nlu-N6AMT by feeding dsRNA from the second instar nymph to adult female, inducing retard development of adult female. In all, we provide the first genome-wide analysis of N6AMT in insects and presented the experimental evidence that N6AMT might have important functions in reproductive development and ovary maturation.

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Improving a method for evaluating alfalfa cultivar resistance to thrips TU Xiong-bing, FAN Yao-li, JI Ming-shan, LIU Zhong-kuan, XIE Nan, LIU Zhen-yu, ZHANG Ze-hua 2016, 15(3): 600-607.  DOI: 10.1016/S2095-3119(15)61197-2 Abstract ( )   PDF in ScienceDirect   The thrips quantity ratio (TQR) model is an important tool for evaluating crop resistance to thrips based on the correlation between thrips quantities and cultivars. Unfortunately, it is inaccurate, and the results appear significantly inconsistent when analysing the same cultivars in the same field study. To improve this model, we first studied the resistance of 28 alfalfa cultivars to thrips in Cangzhou, Hebei Province, north China. The results showed that the most suitable evaluation period was from May to June, as the thrips population was stable during this period. Second, we found that the natural enemy population was significantly positively correlated with the thrips population density (R=0.7275, P<0.0001), which might influence resistance estimation. Hence, we introduced a parameter ‘α’, corresponding to the natural enemy quantity ratio, to eliminate the effect of the natural enemy using “αTQR”. Using the improved method, 28 cultivars were clustered into three classes: the resistant class, sensitive class, and median class. All numerical values were calculated for αTQR displayed as a Gaussian distribution. This information showed that all data should be divided into nine groups using a median value of 1±0.1 with an equal difference of 0.1. Based on the new standard cultivars, Gongnong 1, Alfaking, Cangzhou and Algonquin were classified as highly resistant cultivars; Zhongmu 3, Gongnong 2, Zhongmu 1 and Zhongmu 2 were classified in the resistant group; Queen was classified in the moderately resistant group; Derby, WL354HQ, KRIMA, Apex, 53HR, SARDI 5 and Farmers Treasure were classified in the median class; WL319HQ, WL343HQ and Sitel were classified as the low sensitive group; WL440HQ and SARDI 7 as the moderately sensitive group; WL168HQ and Sanditi as the sensitive group; and SARDI 10, WL363HQ, FD4, WL323 and SOCA as the highly sensitive group.

Animal Science · Veterinary Science

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Organic practices and gender are effective strategies to provide healthy pork loin Javier álvarez-Rodríguez, Daniel Villalba, Dolors Cubiló, Daniel Babot, Marc Tor 2016, 15(3): 608-617.  DOI: 10.1016/S2095-3119(15)61172-8 Abstract ( )   PDF in ScienceDirect   This study evaluated the influence of husbandry (organic feed and management but not free-ranging vs. conventional) and sex (barrows vs. gilts) on pork meat quality. A total of 60 Longissimus thoracis pork muscle samples from different 3-way crossbred genotypes were chosen from 3 conventional and 2 organic pig farms. Technological meat quality was measured at 24 h post-mortem and muscle fatty acid content and composition was analysed by gas chromatography. The loin from organic pigs at 24 h of retail display had lower pH, but it had no detrimental effects on drip loss. All the International Commission on Illumination colour attributes except meat lightness differed between husbandry systems. Moisture and crude protein content were lower whereas intramuscular fat content was greater in organic than in conventional pork. Total saturated fatty acid (SFA), monounsaturated fatty acid (MUFA), polyunsaturated fatty acid (PUFA) and PUFA n-6 contents did not differ between husbandry systems, but total PUFA n-3 (mainly C18:3 n-3) were greater in organic than in conventional pork. Sex did not affect ultimate pH or meat colour attributes but barrows showed lower moisture and greater intramuscular fat than gilts. Total SFA and MUFA content were similar but all the PUFA (both n-6 and n-3) were lower in barrows than in gilts. These results suggest that some bioactive compounds from dietary origin, i.e., linolenic acid (C18:3 n-3) content from dietary vegetable oils (soybean or olive olein), might be used to highlight the nutritive value of (not free-ranging) organic pork meat. In addition, gilts were leaner than barrows and showed a more favourable PUFA/SFA ratio.

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Methylation profile of bovine Oct4 gene coding region in relation to three germ layers ZHOU Xin-yu, LIU Liang-liang, JIA Wen-chao, PAN Chuan-ying 2016, 15(3): 618-628.  DOI: 10.1016/S2095-3119(15)61100-5 Abstract ( )   PDF in ScienceDirect   Previous studies have shown that octamer-binding transcription factor 4 (Oct4) plays a significant role in early embryonic development of mammalian animals, and different Oct4 expression levels induce multi-lineage differentiation which are regulated by DNA methylation. To explore the relationship between the methylation pattern of Oct4 gene exon 1 and embryonic development, in this work, five different tissues (heart, liver, lung, cerebrum and cerebellum) from three germ layers were chosen from low age (50–60 d) and advanced age (60–70 d) of fetal cattle and the differences between tissues or ages were analyzed, respectively. The result showed that the DNA methylation level of Oct4 gene exon 1 was significant different (P<0.01) between any two of three germ layers in low age (<60 d), but kept steady of advanced age (P>0.05) (>60 d), suggesting that 60-d post coital was an important boundary for embryonic development. In addition, in ectoderm (cerebrum and cerebellum), there was no significant methylation difference of Oct4 gene exon 1 between low age and advanced age (P>0.05), but the result of endoderm (liver and lung) and mesoderm (heart) were on the contrary (P<0.01), which indicated the development of ectoderm was earlier than endoderm and mesoderm. The methylation differences from the 3rd, 5th and 9th CpG-dinucleotide loci of Oct4 gene exon 1 were significantly different between each two of three germ layers (P<0.05), indicating that these three loci may have important influence on bovine embryonic development. This study showed that bovine germ layers differentiation was significantly related to the DNA methylation status of Oct4 gene exon 1. This work firstly identified the DNA methylation profile of bovine Oct4 gene exon 1 and its association with germ layers development in fetus and adult of cattle. Moreover, the work also provided epigenetic information for further studying bovine embryonic development and cellular reprogramming.

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Effects of graded fiber level and caecectomy on metabolizable energy value and amino acid digestibility in geese YANG Jing, ZHAI Shuang-shuang, WANG Yong-chang, WANG Shen-shen, YANG Zhi-peng, YANG Lin 2016, 15(3): 629-635.  DOI: 10.1016/S2095-3119(15)61071-1 Abstract ( )   PDF in ScienceDirect   This study was carried out to evaluate the effects of graded levels of fiber from rice hull and the caecectomy on metabolizable energy (ME) and amino acid digestibility (AAD) in 42 Wuzong geese (21 intact and 21 caecectomized). The experiment was a 3×2 factorial randomized design with 3 fiber levels and 2 treatments of geese (caecectomized and intact). Each of the 3 diets was tube-fed (60 g) to 7 caecectomized and 7 intact adult Wuzong geese that had been fasted for 24 h. Excreta were collected for the next 48 h. The same geese were also fasted for estimation of endogenous losses of amino acids (AA) and energy after short period recovery. AA content and metabolizable energy value were determined in the excreta. In general, significant effects were found for fiber levels on apparent digestibility of Thr, Leu, Pro, Glu,Val, Ala and total AA (P<0.05). Apparent metabolizable energy (AME) and true metabolizable energy (TME) were both significantly influenced by increased fiber levels (P<0.01). Compared with caecectomized geese, markedly increases of AME and TME were found in intact geese (P<0.01 for AME and P<0.05 for TME), but no difference in the AAD were found between intact and caecectomized geese (P>0.05), except for Lys and Try (P<0.05). These observations suggested that dietary fiber level played an important role in digestibility of AAD and ME. Intact geese should be chosen for the nutrient evaluation of feedstuffs in practice, resulting from the significant difference between caecectomized and intact geese.

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Cloning, bioinformatics and transcriptional analysis of caffeoylcoenzyme A 3-O-methyltransferase in switchgrass under abiotic stress LIU Si-jia, HUANG Yan-hua, HE Chang-jiu, FANG Cheng, ZHANG Yun-wei 2016, 15(3): 636-649.  DOI: 10.1016/S2095-3119(16)61363-1 Abstract ( )   PDF in ScienceDirect   Genes encoding enzymes involved in the lignin biosynthesis through phenylpropanoid pathway were not only associated with the lignin content, but also related to the abiotic stress resistances. As far as the production of liquid biofuels and cultivation within the marginal land are concerned, switchgrass could be the better candidate to determine the relationship between lower lignin content and physiological function under stress. Caffeoyl-coenzyme A 3-O-methyltransferase (CCoAOMT) is a key enzyme for the methylation reaction of lignin biosynthesis. For this purpose, we cloned a CCoAOMT gene from switchgrass and identified its expression patterns under abiotic stresses. The full-length CCoAOMT gene, designated PvCCoAOMT (GenBank accession no. KF041775), was 1 005-bp in length, has an opening reading frame of 777 nucleotides encoding a 258-amino acid protein. The deduced amino acid sequence of PvCCoAOMT shared a high degree of similarity (up to 98%) with CCoAOMTs from Panicum virgatum allele (BAO20881), Sorghum bicolor (XP002436550) and Zea mays (NP001131288). Using quantitative real-time PCR (qRT-PCR), the significant upregulation of PvCCoAOMT was observed in stem tissues at a later stage (24 h) after drought treatment, with the transcript level increasing 33-fold compared that of the controls. Moderate and insignificant inductions of PvCCoAOMT were also observed in both stems and leaves during the later stages after cold (48 h in stems, 12 h in leaves) and mechanical wounding (48 h in stems, 12 h in leaves) treatments, respectively. Our results showed the different expression patterns of PvCCoAOMT in drought, cold and mechanical wounding stresses. PvCCoAOMT can be highly induced by drought and cold stresses, which indicates that it may play a role in plant abiotic stress resistance, particularly in the regulation of drought and cold resistance.

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Effect of Fuzheng Jiedu granule on immunological function and level of immune-related cytokines in immune-suppressed mice YU Zhan-mei, HUANG Xian-hui, YAN Chao-qun, GAO Jin, LIANG Zi-sen 2016, 15(3): 650-657.  DOI: 10.1016/S2095-3119(14)60971-0 Abstract ( )   PDF in ScienceDirect   Fuzheng Jiedu granule exhibits a number of health benefits and it is thought that the mechanisms involved in these effects are due to the modulation of immunity. In this article, we studied the effect of Fuzheng Jiedu granule on immunological function and the expression of immune-related cytokines in immune-suppressed mice. 72 mice were randomly divided into six groups, with 12 in each group. The control groups included an untreated group, a negative control group (Cyclophosphamide) and a positive control group (Astragalus polysaccharide). There were three treated groups, which were given different doses of Fuzheng Jiedu granule: a low dose (100 mg kg–1), a medium dose (400 mg kg–1) and a high dose (600 mg kg–1). With the exception of the untreated control animals, each group received an intraperitoneal injection of Cyclophosphamide (100 mg kg–1) for 3 days to establish the immune-suppressed model. Mice were then treated for 19 consecutive days and, 24 h after the last treatment, blood was taken for the eyeballs and serum separation was performed. Analysis was made of the levels of related cytokines (IgA, IgG, IgM, IL-6, IFN-γ, C3, C4 and TNF-α), the transformation of lymphocytes and the immune organ indexes. The results showed that Fuzheng Jiedu granule can improve the levels of cytokines, the rate of proliferation of lymphocytes and the immune organ indexes of immune-suppressed mice.

Soil & Fertilization﹒Irrigation﹒Plant Nutrition﹒ Agro-Ecology & Environment

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Effect of chemical and organic fertilization on soil carbon and nitrogen accumulation in a newly cultivated farmland YANG Rong, SU Yong-zhong, WANG Tao, YANG Qin 2016, 15(3): 658-666.  DOI: 10.1016/S2095-3119(15)61107-8 Abstract ( )   PDF in ScienceDirect   Increased food demand from the rapidly growing human population has caused intensive land transition from desert to farmland in arid regions of northwest China. In this developing ecosystem, the optimized fertilization strategies are becoming an urgent need for sustainable crop productivity, efficient resources use, together with the delivery of ecosystems services including soil carbon (C) and nitrogen (N) accumulation. Through a 7-year field experiment with 9 fertilization treatments in a newly cultivated farmland, we tested whether different fertilizations had significant influences on soil C and N accumulation in this developing ecosystem, and also investigated possible mechanisms for this influence. The results showed that applying organic manure in cultivated farmland significantly increased the soil C and N accumulation rates; this influence was greater when it was combined with chemical fertilizer, accumulating 2.01 t C and 0.11 t N ha–1 yr–1 in the most successful fertilization treatment. These high rates of C and N accumulation were found associated with increased input of C and N, although the relationship between the N accumulation rate and N input was not significant. The improved soil physical properties was observed under only organic manure and integrated fertilization treatments, and the significant relationship between soil C or N and soil physical properties were also found in this study. The results suggest that in newly cultivated farmland, long term organic manure and integrated fertilization can yield significant benefits for soil C and N accumulation, and deliver additional influence on physical properties.

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Phosphorus losses via surface runoff in rice-wheat cropping systems as impacted by rainfall regimes and fertilizer applications LIU Jian, ZUO Qiang, ZHAI Li-mei, LUO Chun-yan, LIU Hong-bin, WANG Hong-yuan, LIU Shen, ZOU Guo-yuan, REN Tian-zhi 2016, 15(3): 667-677.  DOI: 10.1016/S2095-3119(15)61087-5 Abstract ( )   PDF in ScienceDirect   Phosphorus (P) losses from agricultural soils contribute to eutrophication of surface waters. This field plot study investigated effects of rainfall regimes and P applications on P loss by surface runoff from rice (Oryza sativa L.) and wheat (Triticum aestivum L.) cropping systems in Lake Taihu region, China. The study was conducted on two types of paddy soils (Hydromorphic at Anzhen site, Wuxi City, and Degleyed at Xinzhuang site, Changshu City, Jiangsu Province) with different P status, and it covered 3 years with low, high and normal rainfall regimes. Four rates of mineral P fertilizer, i.e., no P (control), 30 kg P ha–1 for rice and 20 kg P ha–1 for wheat (P30+20), 75 plus 40 (P75+40), and 150 plus 80 (P150+80), were applied as treatments. Runoff water from individual plots and runoff events was recorded and analyzed for total P and dissolved reactive P concentrations. Losses of total P and dissolved reactive P significantly increased with rainfall depth and P rates (P<0.0001). Annual total P losses ranged from 0.36–0.92 kg ha–1 in control to 1.13–4.67 kg ha–1 in P150+80 at Anzhen, and correspondingly from 0.36–0.48 kg ha–1 to 1.26–1.88 kg ha–1 at Xinzhuang, with 16–49% of total P as dissolved reactive P. In particular, large amounts of P were lost during heavy rainfall events that occurred shortly after P applications at Anzhen. On average of all P treatments, rice growing season constituted 37–86% of annual total P loss at Anzhen and 28–44% of that at Xinzhuang. In both crop seasons, P concentrations peaked in the first runoff events and decreased with time. During rice growing season, runoff P concentrations positively correlated (P<0.0001) with P concentrations in field ponding water that was intentionally enclosed by construction of field bund. The relative high P loss during wheat growing season at Xinzhuang was due to high soil P status. In conclusion, P should be applied at rates balancing crop removal (20–30 kg P ha–1 in this study) and at time excluding heavy rains. Moreover, irrigation and drainage water should be appropriately managed to reduce runoff P losses from rice-wheat cropping systems.

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Impact of temperature on yield and related traits in cotton genotypes Kalim Ullah, Niamatullah Khan, Zahid Usman, Rehmat Ullah, Fazal Yazdan Saleem, Syed Asif Imran Shah, Muhammad Salman 2016, 15(3): 678-683.  DOI: 10.1016/S2095-3119(15)61088-7 Abstract ( )   PDF in ScienceDirect   Cotton growth and development is influenced by various uncontrollable environmental conditions. Temperature variations in the field can be created by planting at different dates. The objective of the present study was to evaluate the effect of planting dates and thermal temperatures (growing degree days) on yield of 4 cotton genotypes, viz., CIM-598, CIM-599, CIM- 602 and Ali Akbar-703. Plants were subjected to 6 planting dates during 2013 and 2014 in a trial conducted in randomized complete block design with four replications. For boll number, boll weight and seed cotton yield, cotton genotypes exhibited significant differences, CIM-599 produced the highest seed cotton yield of 2 062 kg ha–1 on account of maximum boll number and boll weight. The highest seed cotton yield was recorded in planting dates from 15th April to 1st May whereas early and delayed planting reduced the yield due to less accumulation of heat units. Regression analysis revealed that increase of one unit (15 days) from early to optimum date (15th March to 15th April) increased yield by 93.58 kg ha–1. Delay in planting also decreased the seed cotton yield with the same ratio. Thus it is concluded that cotton must be sown from 15th April to 1st May to have good productivity in this kind of environment.

Food Science

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Expression and characterization of a codon-optimized butyrylcholinesterase for analysis of organophosphate insecticide residues TIAN Jing-jing, CHEN Xiang-ning, XIE Yuan-hong, LU Yong, XU Wen-tao, XU Li, DU Bin 2016, 15(3): 684-693.  DOI: 10.1016/S2095-3119(15)61139-X Abstract ( )   PDF in ScienceDirect   Organophosphate insecticide residues on vegetable, fruit, tea and even grains are primary cause of food poisoning. Organophosphate compounds can cause irreversible inhibition of the activity of acetylcholinesterase and butyrylcholinesterase (BChE, EC 3.1.1.8), which are both candidates for rapid detection of organophosphate pesticides. To develop an easy-tohandle method for detecting organophosphate pesticides using BChE, BChE from human was optimized according to the codon usage bias of Pichia pastoris and successfully expressed in P. pastoris GS115. The codon-optimized cDNA shared 37.3% of the codon identity with the native one. However, the amino acid sequence was identical to that of the native human butyrylcholinesterase gene (hBChE) as published. The ratio of guanine and cytosine in four kinds of bases ((G+C) ratio) was simultaneously increased from 40 to 47%. The recombinant hBChE expression reached a total protein concentration of 292 mg mL–1 with an activity of 14.7 U mL–1, which was purified 3.2×103-fold via nickel affinity chromatography with a yield of 68% and a specific activity of 8.1 U mg–1. Recombinant hBChE was optimally active at pH 7.4 and 50°C and exhibited high activity at a wide pH range (>60% activity at pH 4.0 to 8.0). Moreover, it had a good adaptability to high temperature (>60% activity at both 50 and 60°C up to 60 min) and good stability at 70°C. The enzyme can be activated by Li+, Co+, Zn2+ and ethylene diamine tetraacetic acid (EDTA), but inhibited by Mg2+, Mn2+, Fe2+, Ag+ and Ca2+. Na+ had little effect on its activity. The values of hBChE of the Michaelis constant (Km) and maximum reaction velocity (Vm) were 89.4 mmol L–1 and 1 721 mmol min–1 mg–1, respectively. The bimolecular rate constants (Ki) of the hBChE to four pesticides were similar with that of electric eel AChE (EeAChE) and higher than that of horse BChE (HoBChE). All values of the half maximal inhibitory concentration of a substance (IC50) for hBChE were lower than those for HoBChE, but most IC50 for hBChE were lower than those for EeAChE except dichlorvos. The applicability of the hBChE was further verified by successful detection of organophosphate insecticide residues in six kinds of vegetable samples. Thus, hBChE heterologously over-expressed by P. pastoris would provide a sufficient material for development of a rapid detection method of organophosphate on spot and produce the organophosphate detection kit.

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Separation and purification of deoxynivalenol (DON) mycotoxin from wheat culture using a simple two-step silica gel column chromatography ZHAO Xiu-mei, LI Rong-jia, ZHOU Chuang, ZHANG Jie, HE Cheng-hua, ZHENG Ya-ting, WU Wen-da 2016, 15(3): 694-701.  DOI: 10.1016/S2095-3119(15)61098-X Abstract ( )   PDF in ScienceDirect   Deoxynivalenol (DON) is a type B trichothecenes mycotoxin produced by several Fusarium species, often found in foodstuffs for humans and animals. DON is in great demand for the toxicological researches both in vivo and in vitro. In this work, wheat culture was inoculated with a Fusarium graminearum PH-1 strain for DON production. The solvent system for crude extraction was acetonitrile-water (84:16, v/v). A simple two-step silica gel column chromatography was employed to separate the DON mycotoxin from wheat culture, combined with preparative high performance liquid chromatography (preparative HPLC) to purify the compound. The solvent system for the second silica gel column chromatography was methylene chloride-methanol (17:1, v/v), which provided a good elution effect selected on thin layer chromatography (TLC). The target compound was identified by HPLC, and the chemical structure was confirmed by mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. A total of 433 mg of purified DON was obtained from 1 kg of wheat culture, with a purity of 99.01%. The study had provided an easy-operating and cost-effective method to isolate an expensive compound in a simple way.

Short Communication

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Assaying the potential of twenty-one legume plants in Medicago truncatula and M. sativa for candidate model plants for investigation the interactions with Heterodera glycines KONG Ling-an, WU Du-qing, HUANG Wen-kun, PENG Huan, HE Wen-ting, PENG De-liang 2016, 15(3): 702-704.  DOI: 10.1016/S2095-3119(15)61156-X Abstract ( )   PDF in ScienceDirect   Soybean cyst nematode Heterodera glycines is one of the most serious soil-borne pathogens in soybean production. However, the researches were limited in China due to lack of an effective pathosystem. In this study, we screened 21 legume Medicago plants in both Medicago truncatula and Medicago sativa to obtain candidate model plants for establishing a new pathosystem for legume-H. glycines interactions. The nematode infection of tested plants was assayed with Race 3 and 4 respectively, which were two dominant H. glycines inbred races in China soybean producing areas. The results showed that the model legume plant M. truncatula A17 failed to allow Race 3 of H. glycines to complete its life cycle, in contrast, it provided the Race 4 population to form several cyst nematodes, however, the female index (FI) value was approximately 1.6. Three M. sativa cultivars, including Xunlu, Aergangjin and Junren, provided either Race 3 or 4 of H. glycines to develop into mature cysts with their FI value below 5 as well. Our results demonstrated that legume plants in both M. truncatula and M. sativa were not likely to be a model plant for H. glycines because of an extreme high resistance.