Journal of Integrative Agriculture

EDITORAL-Viral pathogens and plant parasitic nematode

Jiban Kumar Kundu

2017, 16(03): 509-509. DOI: 10.1016/S2095-3119(16)61558-7

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This special focus is dedicated to three parts: i) One of the most ubiquitous viral pathogens of stone fruit tree, Plum pox virus (PPV); ii) a re-emerging pathogen, Wheat streak mosaic virus (WSMV) of cereal crops in Central Europe; and iii) a less studied plant parasitic, cyst-forming nematode in cereal crops Heterodera avenae. The pathogens described here are a serious concern for fruit growers and producers of cereals in Europe and elsewhere in the world. This special focus provides the most updated knowledge of the given topics.

Host preference of the major strains of Plum pox virus - opinions based on regional and world-wide sequence data

Nina Sihelská, Miroslav Glasa, Zdeno W ?ubr

2017, 16(03): 510-515. DOI: 10.1016/S2095-3119(16)61356-4

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Plum pox virus (PPV) causes sharka - the most serious viral disease of stone fruit trees. PPV is wide spread in Europe and Mediterranean Basin, its incidence has been further approved in Asia and both Americas. Nine PPV strains have been recognized until now (PPV-D, PPV-M, PPV-Rec, PPV-EA, PPV-C, PPV-T, PPV-W, PPV-CR, and PPV-An), forming molecularly distinct entities, however, only partially differentiable by their biological or epidemiological properties. The most strict virus-host linkages under natural conditions have been detected for strains naturally infecting cherries (PPV-C and PPV-CR). However, although less stringent but still clear host preference is observed also for three epidemiologically most important strains (PPV-D/plum/apricot, PPV-M/peach, and PPV-Rec/plum). So far no genetic marker has been mapped in the PPV genome, which responsibility for the host specificity/preference could be explicitly demonstrated. In this review, we focus on the host preference of three major PPV strains as evidenced by analysis of an extensive dataset of PPV isolates of Slovak and world-wide origin. Together, we discuss several performed relevant experiments and further possible research procedures aimed to better understand the genetic determinants and mechanisms of the host preference of this potyvirus.

Transgenic plum Prunus domestica L., clone C5 (cv. HoneySweet) for protection against sharka disease

Jaroslav Polák, Jiban K Kundu, Boris Kr?ka, Eva Beoni, Petr Komínek, Jitka Pívalová, Jana Jaro?ová

2017, 16(03): 516-522. DOI: 10.1016/S2095-3119(16)61491-0

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Sharka caused by Plum pox virus (PPV) is the most harmful disease of stone fruits in Europe and elsewhere in the world. There is no highly PPV resistant cultivar of plum. Biotech approach has led to the development of resistance through genetic engineering. In this study, we evaluated a transgenic plum Prunus domestica L., clone C5 (cv. HoneySweet), where the PPV resistance is based on RNA interference (RNAi). Resistance in C5 plums has been evaluated for PPV, Prune dwarf virus (PDV), and Apple chlorotic leaf spot virus (ACLSV) in a regulated field trial in the Czech Republic for 12 years (2002–2013). Even under high and permanent infection pressure introduced through graft inoculation of the viruses, PPV has been detected in C5 trees only in several leaves situated close to the point of inoculum grafting in the first nine years. Mild symptoms of PPV disappeared year by year. No PPV symptoms were observed in the last three years and results of ELISA detection were negative. Similar results were obtained, when RT-PCR was used for PPV detection. No natural infection of PPV by aphids was recorded in C5 plums, even if PPV infected plants were growing in the same experimental field in close vicinity. Co-infections of PPV with PDV and/or ACLSV had practically no influence on stability of resistance in C5 trees. Twelve years of field testing in the Czech Republic clearly demonstrated the high level of resistance of C5 either to PPV infection by graft inoculation or natural infection through aphid vectors.

Wheat streak mosaic virus: incidence in field crops, potential reservoir within grass species and uptake in winter wheat cultivars

Jana Chalupniková, Jiban Kumar Kundu, Khushwant Singh, Pavla Bartaková, Eva Beoni

2017, 16(03): 523-531. DOI: 10.1016/S2095-3119(16)61486-7

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Wheat streak mosaic virus (WSMV) has become a re-emerging pathogen in cereal crops in the Czech Republic. WSMV was first reported in the former Czechoslovakia in the early 1980s, and then no record of the virus was documented until 2009. The incidence of the virus was recorded in recent years in several winter wheat fields and many grass species. Here, we surveyed the incidence of WSMV in cereal crops. The results demonstrated the existence of the virus in winter wheat and volunteer wheat during each year of the monitoring period, which spanned from 2013–2016. Although the range of infected samples was low (6.4% of the total tested samples), a high incidence of well-distributed virus was recorded. In at least six fields, the virus reached severe and potentially epidemic levels. In accordance with our previous report detailing WSMV infection of native grasses, we tested several grass species commonly grown in the Czech Republic. We found that some grass species acted as experimental hosts and possible reservoirs of the virus; these included Anthoxanthum odoratum (sweet vernal grass), Arrhenatherum elatius (false oat-grass), Lolium multiflorum (Italian rye-grass), Bromus japonicus (Japanese chess), Echinochloa crus-galli (barnyard grass), Holcus lanatus (meadow soft grass) and Holcus mollis (creeping soft grass). Some of these grass species are also important weeds of cereals, which may be the potential source of WSMV infection in cereal crops. Several widely used winter wheat cultivars were tested in the field after artificial inoculation with WSMV to evaluate virus titre by RT-qPCR. Overall, the tested cultivars had a low virus titre, which is associated with mild disease symptoms and may provide a good level of crop resistance to WSMV.

Morphological and molecular characterizations of cereal cyst nematode Heterodera avenae Wollenweber, 1924 from the Czech Republic

Shesh Kumari

2017, 16(03): 532-539. DOI: 10.1016/S2095-3119(16)61485-5

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The cereal cyst nematode, Heterodera avenae Wollenweber, 1924, is a major pest of cereal crops throughout the world and causes serious yield losses, especially of wheat. Previous studies have shown that this species is widely distributed in the Czech Republic. In this study, seven populations of H. avenae were molecularly studied, and one population was morphologically described. Three regions (18S, 28S, and internal transcribed spacer 1) of ribosomal DNA were sequenced and the analysis of the 18S gene of six populations did not reveal any variation, whereas the internal transcribed spacer 1 and 28S sequences of six populations differed by only two nucleotides from a population in ?ilina. Precise and quick identification of cereal cyst nematodes is important for effective control measures and ribosomal sequence analyses of seven populations in this study will be useful in future phylogenetic studies of Heterodera spp. occurring in the Czech Republic.

Fine mapping of powdery mildew resistance gene PmTm4 in wheat using comparative genomics

XIE Jing-zhong, WANG Li-li, WANG Yong, ZHANG Huai-zhi, ZHOU Sheng-hui, WU Qiu-hong, CHEN Yong-xing, WANG Zhen-zhong, WANG Guo-xin, ZHANG De-yun, ZHANG Yan, HU Tie-zhu, LIU Zhi-yong

2017, 16(03): 540-550. DOI: 10.1016/S2095-3119(16)61377-1

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Powdery mildew, caused by Blumeria graminis f. sp. tritici, is one of the most severe wheat diseases. Mining powdery mildew resistance genes in wheat cultivars and their appliance in breeding program is a promising way to control this disease. Genetic analysis revealed that a single dominant resistance gene named PmTm4 originated from Chinese wheat line Tangmai 4 confers resistance to prevailing isolates of B. graminis f. sp. tritici isolate E09. Detailed comparative genomics analyses helped to develop closely linked markers to PmTm4 and a fine genetic map was constructed using large F₂ population, in which PmTm4 was located into a 0.66-cM genetic interval. The orthologous subgenome region of PmTm4 in Aegilops tauschii was identified, and two resistance gene analogs (RGA) were characterized from the corresponding sequence scaffolds of Ae. tauschii draft assembly. The closely linked markers and identified Ae. tauschii orthologs in the mapping interval provide an entry point for chromosome landing and map-based cloning of PmTm4.

Development of glyphosate-tolerant transgenic cotton plants harboring the G2-aroA gene

ZHANG Xiao-bing, TANG Qiao-ling, WANG Xu-jing, WANG Zhi-xing

2017, 16(03): 551-558. DOI: 10.1016/S2095-3119(16)61458-2

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Given that glyphosate weed control is an effective strategy to reduce costs and improve economic outcomes of agricultural production in China, the development of glyphosate-resistant cotton holds great promise. Using an Agrobacterium-mediated transformation method, a new G2-aroA gene that encodes 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) was transformed into cotton cultivar K312. The transgenic cotton plants were regenerated from a callus tissue culture via kanamycin selection. Ten regenerated cotton plants were obtained and allowed to flower normally to produce fruit. The results from polymerase chain reaction (PCR) and Southern and Western blot analyses indicated that the target gene was integrated into the cotton chromosome and was expressed effectively at the protein level. The glyphosate tolerance analysis showed that the transgenic cotton had a high resistance to glyphosate. Further, even cotton treated with 45.0 mmol L^–1 of glyphosate was able to slowly grow, bloom and seed. The transgenic cotton may be used for cotton breeding research of glyphosate-tolerant cotton.

The NAC-like transcription factor SiNAC110 in foxtail millet (Setaria italica L.) confers tolerance to drought and high salt stress through an ABA independent signaling pathway

XIE Li-na, CHEN Ming, MIN Dong-hong, FENG Lu, XU Zhao-shi, ZHOU Yong-bin, XU Dong-bei, LI Lian-cheng, MA You-zhi, ZHANG Xiao-hong

2017, 16(03): 559-571. DOI: 10.1016/S2095-3119(16)61429-6

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Foxtail millet (Setaria italica (L.) P. Beauv) is a naturally stress tolerant crop. Compared to other gramineous crops, it has relatively stronger drought and lower nutrition stress tolerance traits. To date, the scope of functional genomics research in foxtail millet (S. italic L.) has been quite limited. NAC (NAM, ATAF1/2 and CUC2)-like transcription factors are known to be involved in various biological processes, including abiotic stress responses. In our previous foxtail millet (S. italic L.) RNA seq analysis, we found that the expression of a NAC-like transcription factor, SiNAC110, could be induced by drought stress; additionally, other references have reported that SiNAC110 expression could be induced by abiotic stress. So, we here selected SiNAC110 for further characterization and functional analysis. First, the predicted SiNAC110 protein encoded indicated SiNAC110 has a conserved NAM (no apical meristem) domain between the 11–139 amino acid positions. Phylogenetic analysis then indicated that SiNAC110 belongs to subfamily III of the NAC gene family. Subcellular localization analysis revealed that the SiNAC110-GFP fusion protein was localized to the nucleus in Arabidopsis protoplasts. Gene expression profiling analysis indicated that expression of SiNAC110 was induced by dehydration, high salinity and other abiotic stresses. Gene functional analysis using SiNAC110 overexpressed Arabidopsis plants indicated that, under drought and high salt stress conditions, the seed germination rate, root length, root surface area, fresh weight, and dry weight of the SiNAC110 overexpressed lines were significantly higher than the wild type (WT), suggesting that the SiNAC110 overexpressed lines had enhanced tolerance to drought and high salt stresses. However, overexpression of SiNAC110 did not affect the sensitivity of SiNAC110 overexpressed lines to abscisic acid (ABA) treatment. Expression analysis of genes involved in proline synthesis, Na⁺/K⁺ transport, drought responses, and aqueous transport proteins were higher in the SiNAC110 overexpressed lines than in the WT, whereas expression of ABA-dependent pathway genes did not change. These results indicated that overexpression of SiNAC110 conferred tolerance to drought and high salt stresses, likely through influencing the regulation of proline biosynthesis, ion homeostasis and osmotic balance. Therefore SiNAC110 appears to function in the ABA-independent abiotic stress response pathway in plants.

Evaluation of parameters affecting Agrobacterium-mediated transient expression in citrus

LI Fang, DAI Su-ming, DENG Zi-niu, LI Da-zhi, LONG Gui-you, LI Na, LI Yi, Alexandra Gentile

2017, 16(03): 572-579. DOI: 10.1016/S2095-3119(16)61460-0

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Agrobacterium-mediated transient expression assays are a convenient alternative to stable expression because they are simple, easy to perform, and achieve gene expression rapidly. This study investigated the factors affecting transient gene expression efficiency in citrus by observing the cryo-sectioning of leaf samples under a laser confocal microscope. These factors included the composition of the infiltration buffer, the Agrobacterium cell density, the leaf development stage, the incubation temperature, and plant genotype. The highest transient expression level of yellow fluorescent protein (YFP) was detected in Mexican lime (Citrus aurantifolia) on the third day after the intermediate-aged leaves were infiltrated with the improved infiltration buffer 1 (15 mmol L^-1 2-(N-morpholino) ethanesulfonic acid, 10 mmol L^-1 MgCl₂, and 200 μmol L^-1 acetosyringone), which had an optical density of 0.8 and was incubated at 22°C. Additionally, this transient expression assay was applied to other citrus genotypes. Of note, trifoliate orange (Poncirus trifoliata) and kumquat (Fortunella obovate) had higher expression efficiency than other six genotypes of the Citrus genus. Our study provides research basis for the selection of optimization strategies in transient gene expression and improves the method for available genome investigation in citrus.

Expression and functional analysis of FaPHO1;H9 gene of strawberry (Fragaria×ananassa)

CAO Fei, LI He, WANG Shou-ming, LI Xiao-ming, DAI Hong-yan, ZHANG Zhi-hong

2017, 16(03): 580-590. DOI: 10.1016/S2095-3119(16)61433-8

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Although the phosphate 1 (PHO1) gene family has been implicated in inorganic phosphate transport and homeostasis, the underlying mechanism of this gene in the strawberry has not yet been revealed. In the present study, we analyzed the expression of the PHO1;H9 gene in the strawberry (Fragaria×ananassa), revealing the involvement of this gene in the regulation of phosphorus (P) content. The coding sequence (CDS) of the PHO1;H9 gene, was isolated from the cultivated strawberry ‘Sachinoka’ and named as FaPHO1;H9. The full-length CDS of this gene was 2 292 bp, encoding 763 amino acids, and the protein contained both SYG1/Pho81/XPR1 (SPX) and ERD1/ XPR1/SYG1 (EXS) domains, which were involved in phosphate (Pi) signaling. Real-time reverse transcription-polymerase chain reaction (RT-PCR) data suggested that the level of FaPHO1;H9 expression was consistent with the P content in different organs, except for the petiole. Particularly, its expression level was also correlated with P content in fruits of different developmental stages. The expression of FaPHO1;H9 was also consistent with P content in leaves under different concentrations of P fertilizer application. Furthermore, transgenic Arabidopsis lines were generated, and the P content in Arabidopsis plants over-expressing FaPHO1;H9 was significantly higher than that in wild-type plants. Therefore, we proposed that FaPHO1;H9 functions in P transport.

High-throughput sequencing of highbush blueberry transcriptome and analysis of basic helix-loop-helix transcription factors

SONG Yang, LIU Hong-di, ZHOU Qiang, ZHANG Hong-jun, ZHANG Zhi-dong, LI Ya-dong, WANG Hai-bo, LIU Feng-zhi

2017, 16(03): 591-604. DOI: 10.1016/S2095-3119(16)61461-2

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The highbush blueberry (Vaccinium corymbosum), Duke, was used to construct a de novo transcriptome sequence library and to perform data statistical analysis. Mega 4, CLC Sequence Viewer 6 software, and quantitative PCR were employed for bioinformatics and expression analyses of the basic helix-loop-helix (BHLH) transcription factors of the sequencing library. The results showed that 28.38 gigabytes of valid data were obtained from transcriptome sequencing and were assembled into 108 033 unigenes. Functional annotation showed that 32 244 unigenes were annotated into Clusters of Orthologous Groups (COG) and Gene Ontology (GO) databases, whereas the rest of the 75 789 unigenes had no matching information. By using COG and GO classification tools, sequences with annotation information were divided into 25 and 52 categories, respectively, which involved transport and metabolism, transcriptional regulation, and signal transduction. Analysis of the transcriptome library identified a total of 59 BHLH genes. Sequence analysis revealed that 55 genes of that contained a complete BHLH domain. Furthermore, phylogenetic analysis showed that BHLH genes of blueberry (Duke) could be divided into 13 sub-groups. PCR results showed that 45 genes were expressed at various developmental stages of buds, stems, leaves, flowers, and fruits, suggesting that the function of BHLH was associated with the development of different tissues and organs of blueberry, Duke. The present study would provided a foundation for further investigations on the classification and functions of the blueberry BHLH family.

Effect of seed priming with different concentrations of potassium nitrate on the pattern of seed imbibition and germination of rice (Oryza sativa L.)

Anisa Ruttanaruangboworn, Wanchai Chanprasert, Pitipong Tobunluepop, Damrongvudhi Onwimol

2017, 16(03): 605-613. DOI: 10.1016/S2095-3119(16)61441-7

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Low germination and vigor of rice seed associated with dry-seed broadcasting are common problems encountered by rice growers. The objectives of this study were to evaluate the role of potassium nitrate (KNO₃) on the pattern of seed imbibition and to determine the effect of seed priming with KNO₃ on the germination percentage, speed and uniformity of germination in rice seed. Experiment 1 compared the patterns of seed imbibition of six concentrations of KNO₃ (0, 0.25, 0.50, 1.00, 1.50, and 2.00%) in two rice cultivars - KDML105 and RD15. The results showed that soaking rice seed in KNO₃ at higher concentrations could delay the imbibition time. The higher concentrations of KNO₃ delayed the imbibition time of rice seed and took a longer time to reach the end of phases 1 and 2 compared to the lower concentrations. The patterns of seed imbibition using distilled water of both rice cultivars (KDML105 and RD15) were quite similar, but with different concentrations of KNO₃, the imbibition time taken to reach the end of phases 1 and 2 was slightly postponed in KDML105 suggesting that different rice cultivars may need different imbibition times for soaking seed in the priming process. Experiment 2 evaluated the effects of seed priming with 1.0 and 2.0% KNO₃at different imbibition times. It was found that priming with 1.0% KNO₃ showed better seed germination than priming with 2.0% KNO₃ and seed priming with 1.0% KNO₃at the imbibition time of early phase 2 (or 28 h for KDML105) improved seed germination and increased both the speed and uniformity of seed germination. The results of this study show promise for the use of priming with 1.0% KNO₃ soaked until early phase 2 of seed imbibition for improving the seed germination and vigor of rice in dry seed broadcasting.

Grain yield and water use of winter wheat as affected by water and sulfur supply in the North China Plain

XIE Ying-xin, ZHANG Hui, ZHU Yun-ji, ZHAO Li, YANG Jia-heng, CHA Fei-na, LIU Cao, WANG Chen-yang, GUO Tian-cai

2017, 16(03): 614-625. DOI: 10.1016/S2095-3119(16)61481-8

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Water shortage has threatened sustainable development of agriculture globally as well as in the North China Plain (NCP). Irrigation, as the most effective way to increase food production in dry land, may not be readily available in the situation of drought. One of the alternatives is to supply plants with enough nutrients so that they can be more sustainable to the water stress. The objective of this study was to explore effects of irrigation and sulphur (S) application on water consumption, dry matter accumulation (DMA), and grain yield of winter wheat in NCP. Three irrigation regimes including no irrigation (rainfed, I₀) during the whole growth period, once irrigation only at jointing stage (90 mm, I₁), and twice respective irrigation at jointing and anthesis stages (90 mm plus 90 mm, I₂), and two levels of S application including 0 (S₀) and 60 kg ha^–1(S₆₀) were designed in the field experiment in NCP. Results showed that increasing irrigation times significantly increased mean grain yield of wheat by 12.5–23.7% and nitrogen partial factor productivity (NPFP) by 21.2–45.0% in two wheat seasons, but markedly decreased crop water use efficiency (YWUE). Furthermore, S supply 60 kg ha^–1 significantly increased mean grain yield, YWUE, IWUE and NPFP by 5.6, 6.1, 23.2, and 5.6% (across two wheat seasons), respectively. However, we also found that role of soil moisture prior to S application was one of important greater factors on improving the absorption and utilization of storage water and nutrients of soil. Thus, water supply is still the most important factor to restrict the growth of wheat in the present case of NCP, supplying 60 kg ha^–1 S with once irrigation 90 mm at the jointing stage is a relatively appropriate recommended combination to improve grain yield and WUE of wheat when saving water resources is be considered in irrigated wheat farmlands of NCP.

Top-grain filling characteristics at an early stage of maize (Zea mays L.) with different nitrogen use efficiencies

SHEN Li-xia, HUANG Yan-kai, LI Ting

2017, 16(03): 626-639. DOI: 10.1016/S2095-3119(16)61457-0

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Maize genotypes vary significantly in their nitrogen use efficiencies (NUEs). Better understanding of early grain filling characteristics of maize is important, especially for maize with different NUEs. The objectives of this research were (i) to investigate the difference in apical kernel development of maize with different NUEs, (ii) to determine the reaction of apical kernel development to N application levels, and (iii) to evaluate the relationship between apical kernel development and grain yield (GY) for different genotypes of maize. Three maize hybrid varieties with different NUEs were cultivated in a field with different levels of N fertilizer arranged during two growing seasons. Kernel fresh weight (KFW), volume (KV) and dry weight (KDW) of apical kernel were evaluated at an early grain filling stage. Ear characteristics, GY and its components were determined at maturity stage. Apical kernel of the high N and high efficiency (HN-HE) type (under low N, the yield is lower, and under higher N, the yield is higher) developed better under high N (N210 and N240, pure N of 210 and 240 kg ha^–1) than at low N (N120 and N140, pure N of 120 and 140 kg ha^–1). The low N and high efficiency (LN-HE) type (under low N, the yield is higher, while under higher N, the yield is not significantly higher) developed better under low N than at high N. The double high efficiency (D-HE) type (for both low and high N, the yield is higher) performed well under both high and low N. Apical kernel reacted differently to the N supply. Apical kernel developed well at an early grain filling stage and resulted in a higher kernel number (KN), kernel weight (KW) and GY with better ear characteristics at maturity.

Effect of replacing antibiotics using multi-enzyme preparations on production performance and antioxidant activity in piglets

HAN Xin-yan, YAN Feng-ying, NIE Xin-zheng, XIA Wei, CHEN Sha, ZHANG Xiao-xu, QIAN Li-chun

2017, 16(03): 640-647. DOI: 10.1016/S2095-3119(16)61425-9

Abstract ( )

The study was conducted to investigate the effects of replacing antibiotics using multi-enzyme preparations on growth performance, coefficient of total tract apparent digestibility, digestive enzyme activity, and antioxidant property in piglets. A total of 160 piglets ((21.35±0.22) kg) were randomly assigned to five dietary treatments: 1) basal diet supplemented with antibiotics (AC), 2) antibiotic diet supplemented with 0.5 g kg^–1 multi-enzyme preparations (AC+0.5EP), 3) antibiotic diet supplemented with 1.5 g kg^–1 multi-enzyme preparations (AC+1.5EP), 4) basal diet supplemented with a half dosage of antibiotics and 1.5 g kg^–1 multi-enzyme preparations (AH+1.5EP), and 5) basal diet supplemented with 1.5 g kg^–1 multi-enzyme preparations (BC+1.5EP). The results showed that AC+1.5EP significantly improved the feed efficiency, apparent digestibility of ether extract (EE) and crude ash (CA), lipase activity in pancreas and duodenum content, maltase and lactase activity in jejunum and ileum mucosa, glutathione peroxidase (GSH-Px) concentration in serum and liver, and decreased malondialdehyde (MDA) concentration in serum and liver compared with piglets receiving AC (P<0.05). Piglets receiving BC+1.5EP showed no significant difference in growth performance (P>0.05) but had lower MDA concentration than piglets receiving AC (P<0.05). The apparent digestibility of EE and crude fiber (CF), duodenal lipase activity, jejunum mucosa maltase, and ileum mucosa lactase activity of piglets receiving AH+1.5EP or BC+1.5EP were significantly improved compared with piglets receiving AC (P<0.05). These results indicated an additive growth promotion effect between antibiotics and multi-enzyme preparations on piglets, and the multi-enzyme preparations may be used as substitutes for antibiotics for improving piglet production performance and health status.

Dietary arginine supplementation in multiparous sows during lactation improves the weight gain of suckling piglets

ZHU Cui, GUO Chang-yi, GAO Kai-guo, WANG Li, CHEN Zhuang, MA Xian-yong, JIANG Zong-yong

2017, 16(03): 648-655. DOI: 10.1016/S2095-3119(16)61426-0

Abstract ( )

This study investigated the effects of dietary arginine (Arg) supplementation, just during lactation, on sow and litter performance, plasma concentrations of metabolites and hormones, and milk yield and composition in multiparous sows. Thirty-one sows were randomly assigned to 3 dietary treatments supplemented with 0.0 (control, n=10), 0.5% (n=10), or 1.0% (n=11) L-Arg-HCl, respectively. Experimental diets were provided to the sows from d 3 to 21 of lactation. Plasma and milk samples were collected at d 14 and 21 of lactation. The average daily gain (ADG) of piglets from sows fed diets supplemented with 0.5 or 1.0% L-Arg-HCl at d 3 to 14 of lactation, were higher than that of controls (P<0.05). Maternal supplementation with 1.0% L-Arg-HCl also increased ADG of piglets between d 3 and 21 of lactation than that of the controls (P<0.05). There was no significant effect of supplementation on average daily feed intake (ADFI), body weight loss, and backfat thickness loss of lactating sows. Supplementation with 0.5 or 1.0% L-Arg-HCl had a trend towards increasing milk yields and milk fat contents (0.05<P<0.10); milk protein and lactose were unchanged. Supplementation with 1.0% L-Arg-HCl increased plasma concentrations of prolactin and insulin in sows at d 14 and 21 of lactation, and plasma concentrations of non-esterified fatty acid (NEFA), insulin-like growth factor-1 (IGF-1), and nitric oxide (NO) in sows at d 21 of lactation, when compared to the controls (P<0.05). Supplementation with 1.0% L-Arg-HCl increased IGF-1 and spermine in milk at d 14 of lactation, relative to the controls (P<0.05). Plasma Arg concentrations at d 14 and 21 of lactation, as well as plasma NO level and milk IGF-1 at d 21 of lactation, were increased, while plasma urea nitrogen (PUN) concentration at d 14 and 21 of lactation was decreased, by supplementation with 0.5 or 1.0% L-Arg-HCl when compared to the controls (P<0.05). Collectively, dietary supplementation of multiparous sows with Arg, just during lactation, is beneficial for enhancing litter weight gain but the complete mechanism remains to be determined and may involve in the maternal endocrine changes and milk polyamines contents.

The genetic diversity analysis in the donkey myostatin gene

LIU Dong-hua, HAN Hao-yuan, ZHANG Xin, SUN Ting, LAN Xian-yong, CHEN Hong, LEI Chu-zhao, DANG Rui-hua

2017, 16(03): 656-663. DOI: 10.1016/S2095-3119(16)61445-4

Abstract ( ) PDF in ScienceDirect

Myostatin (MSTN) gene negatively controls skeletal muscle development and growth, variations of which play an important role in the regulation of skeletal muscle growth in mammals. However, study on genetic polymorphism of MSTN gene in donkey is limited. In this study, we screened the single nucleotide polymorphsims (SNPs) of MSTN gene in 13 Chinese donkey breeds. Four novel SNPs (g.229T>C, g.872A>G, g.2014G>A, and g.2395C>G) were detected and genotyped by sequencing and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) methods. Six haplotypes (H1–H6) were analyzed, which indicated abundant haplotype diversities in Chinese donkeys. The haplotype H1 was the most dominant and ancient in all breeds. Xinjiang donkey displayed the highest haplotype diversity. The Neighbour-Joining (NJ) tree of MSTN gene among different species was constructed. The clustering result of nine species was consistent with the fact of species differentiation. Our results will provide a reliable theoretical basis for the preservation, exploration and utilization of Chinese donkey genetic resources.

Effects of molasses on the fermentation characteristics of mixed silage prepared with rice straw, local vegetable by-products and alfalfa in Southeast China

WANG Jian, CHEN Lei, YUAN Xian-jun, GUO Gang, LI Jun-feng, BAI Yun-feng, SHAO Tao

2017, 16(03): 664-671. DOI: 10.1016/S2095-3119(16)61473-9

Abstract ( ) PDF in ScienceDirect

This experiment was conducted to study the effect of molasses on the fermentation characteristics of mixed silage ensiled rice straw and vegetable by-products with alfalfa. Mixture (202 g kg^–1 dry matter (DM)) consisting of rice straw, broccoli residue and alfalfa at the ratio of 5:4:1 was ensiled with three experimental treatments: (1) no additives (control); (2) molasses at 2.5% (M1); (3) molasses at 5% (M2) on a fresh matter basis of mixture, respectively. All treatments were packed into laboratory-scale silos, and three silos per treatment were sampled on days 1, 3, 5, 14 and 30. The result showed that the pH value of all mixed silages decreased gradually with the time of ensiling except for the control silage, in which a significant increase (P<0.05) on day 30 occurred. The lactic acid content increased gradually with the time of ensiling and reached the highest value on day 14, and a marked decrease (P<0.05) was found in the control silage on day 30. The changes of acetic acid content showed similar pattern with lactic acid content. A trace amount of propionic and butyric acid contents were found in the three mixed silages during the fermentation period. Comparing to the control, M1 and M2 treatments improved the fermentation quality of mixed silages as indicated by higher (P<0.05) lactic acid contents and lower (P<0.05) pH and ammonia-N contents. The Flieg points also showed that M1 and M2 silages were well preserved, whereas the control silage had a bad quality. Overall, the findings of this study suggested that adding molasses could improve fermentation quality of mixed silage, and M1 was more suitable for practical application.

Acute and subchronic toxicity as well as evaluation of safety pharmacology of modified pulsatilla granules

JIA Rui-lin, SONG Xu, GUO Yu-fei, YIN Zhong-qiong, LIU Fei, XIONG Juan, LIU Qiu-yan, JIA Ren-yong, LI Li-xia, ZOU Yuan-feng, YIN Li-zi, HE Chang-liang, LIANG Xiao-xia, YUE Gui-zhou

2017, 16(03): 671-678. DOI: 10.1016/S2095-3119(16)61401-6

Abstract ( ) PDF in ScienceDirect

The present study investigated acute and subchronic toxicity and safety pharmacology of modified pulsatilla granules (MPG) to provide a basis for a comprehensive understanding of MPG toxicity. The results of acute toxicity testing showed that the median lethal dose of MPG was more than 5 000 mg kg^–1, suggesting that MPG was considered as practically non-toxic. The subchronic toxicity study for 30 days was conducted by daily oral administration at doses of 375, 750 and 1 500 mg kg^–1 in Sprague-Dawley rats. The results of subchronic toxicity study showed that the body weight and relative organ weight were not significantly changed by administration of MPG. The clinical chemistry study showed that MPG could induce kidney and liver damages. In histopathological, mild lesions in liver and kidney were also observed, suggesting that the liver and kidney might be potential target organs of MPG. In the safety pharmacology study, MPG did not exhibited any side effects to rats in cardiovascular system, respiratory system and central nervous system. These results suggested that MPG could be considered safe for veterinary use.

Effects of reclaimed water irrigation and nitrogen fertilization on the chemical properties and microbial community of soil

GUO Wei, Mathias N Andersen, QI Xue-bin, LI Ping, LI Zhong-yang, FAN Xiang-yang, ZHOU Yuan

2017, 16(03): 679-690. DOI: 10.1016/S2095-3119(16)61391-6

Abstract ( ) PDF in ScienceDirect

The ecological effect of reclaimed water irrigation and fertilizer application on the soil environment is receiving more attention. Soil microbial activity and nitrogen (N) levels are important indicators of the effect of reclaimed water irrigation on environment. This study evaluated soil physicochemical properties and microbial community structure in soils irrigated with reclaimed water and receiving varied amounts of N fertilizer. The results indicated that the reclaimed water irrigation increased soil electrical conductivity (EC) and soil water content (SWC). The N treatment has highly significant effect on the ACE, Chao, Shannon (H) and Coverage indices. Based on a 16S ribosomal RNA (16S rRNA) sequence analysis, the Proteobacteria, Gemmatimonadetes and Bacteroidetes were more abundant in soil irrigated with reclaimed water than in soil irrigated with clean water. Stronger clustering of microbial communities using either clean or reclaimed water for irrigation indicated that the type of irrigation water may have a greater influence on the structure of soil microbial community than N fertilizer treatment. Based on a canonical correspondence analysis (CCA) between the species of soil microbes and the chemical properties of the soil, which indicated that nitrate N (NO₃^–-N) and total phosphorus (TP) had significant impact on abundance of Verrucomicrobia and Gemmatimonadetes, meanwhile the pH and organic matter (OM) had impact on abundance of Firmicutes and Actinobacteria significantly. It was beneficial to the improvement of soil bacterial activity and fertility under 120 mg kg^–1 N with reclaimed water irrigation.

Antioxidant compounds and minerals in tomatoes by Trichoderma-enriched biofertilizer and their relationship with the soil environments

Md. Yeakub Khan, Md. Manjurul Haque, Abul Hossain Molla, Md. Mizanur Rahman, Mohammad Zahangeer Alam

2017, 16(03): 691-703. DOI: 10.1016/S2095-3119(16)61350-3

Abstract ( ) PDF in ScienceDirect

The experiment was conducted to evaluate the efficacy of Trichoderma-enriched biofertilizer (BioF/compost) on antioxidants and minerals in ripe tomatoes and soil health improvements in terms of nutrient availability and microbial populations. The study was comprised of six treatments: control (zero input); recommended doses of NPK (135.5, 45.6 and 22.9 kg ha^–1, respectively); 100% BioF/compost; 75% BioF/compost+25% N; 50% BioF/compost+50% N; and 25% BioF/compost+75% N. The recommended doses of P and K were used in the last three treatments. It was found that the application of 100% BioF/compost enhanced plant growth, leaf greenness, and produced 12.9% higher yield compared to the recommended doses of NPK and other treatments. Mineral contents (P, K, Ca, Mg, Cu, Fe, Mn and Zn) in tomato roots, shoots and fruits and antioxidant compounds, i.e., ascorbic acid, β-carotine, and lycopene were increased significantly in fruits fertilized with 100% BioF/compost. The high efficiency of Trichoderma compost might be the result of its potential of nutrient solubilization and harboring soil microorganisms. Collectively, BioF/compost increased soil fertility and favored growth of microbes in the rhizosphere which ultimately contributed to higher yield, antioxidant, and mineral concentrations in tomatoes. Thus, Trichoderma-enriched biofertilizer may reduce application of chemical fertilizers and therefore, can be considered as a noble practice in sustainable agriculture.

Effects on soil quality of biochar and straw amendment in conjunction with chemical fertilizers

HE Li-li, ZHONG Zhe-ke, YANG Hui-min

2017, 16(03): 704-712. DOI: 10.1016/S2095-3119(16)61420-X

Abstract ( ) PDF in ScienceDirect

The objective of this study was to evaluate the effects on chemical and microbiological properties of paddy soil of short-term biochar, straw, and chemical fertilizers compared with chemical fertilization alone. Five soil fertilization treatments were evaluated: regular chemical fertilizers (RF), straw+regular chemical fertilizers (SRF), straw biochar+regular chemical fertilizers (SCRF), bamboo biochar (BC)+regular chemical fertilizers (BCRF), and straw biochar+70% regular chemical fertilizers (SC+70%RF). Their effects were investigated after approximately 1.5 years. The soil pH and cation exchange capacity (CEC) were significantly higher in biochar-treated soils. The soil phosphorous (P) and potassium (K) contents increased with biochar application. The soil Colwell P content was significantly increased with the addition of straw biochar in the treatments of SCRF and SC+70%RF. The oxygen (O):carbon (C) ratio doubled in BC picked from the soil. This indicated that BC underwent a significant oxidation process in the soil. The denaturing gradient gel electrophoresis (DGGE) fingerprints of microbial communities differed among the treatments. Soils with added biochar had higher Shannon diversity and species richness indices than soils without biochars. The results suggest that biochar can improve soil fertility.

Corn cob biochar increases soil culturable bacterial abundance without enhancing their capacities in utilizing carbon sources in Biolog Eco-plates

JIANG Lin-lin, HAN Guang-ming, LAN Yu, LIU Sai-nan, GAO Ji-ping, YANG Xu, MENG Jun, CHEN Wen-fu

2017, 16(03): 713-724. DOI: 10.1016/S2095-3119(16)61338-2

Abstract ( ) PDF in ScienceDirect

Biochar has been shown to influence soil microbial communities in terms of their abundance and diversity. However, the relationship among microbial abundance, structure and C metabolic traits is not well studied under biochar application. Here it was hypothesized that the addition of biochar with intrinsic properties (i.e., porous structure) could affect the proliferation of culturable microbes and the genetic structure of soil bacterial communities. In the meantime, the presence of available organic carbon in biochar may influence the C utilization capacities of microbial community in Biolog Eco-plates. A pot experiment was conducted with differenct biochar application (BC) rates: control (0 t ha^–1), BC1 (20 t ha^–1) and BC2 (40 t ha^–1). Culturable microorganisms were enumerated via the plate counting method. Bacterial diversity was examined using denaturing gradient gel electrophoresis (DGGE). Microbial capacity in using C sources was assessed using Biolog Eco-plates. The addition of biochar stimulated the growth of actinomyces and bacteria, especially the ammonifying bacteria and azotobacteria, but had no significant effect on fungi proliferation. The phylogenetic distribution of the operational taxonomic units could be divided into the following groups with the biochar addition: Firmicutes, Acidobacteria, Gemmatimonadetes, Actinobacteria, Cyanobacteria and α-, β-, γ- and δ-Proteobacteria (average similarity >95%). Biochar application had a higher capacity utilization for L-asparagine, Tween 80, D-mannitol, L-serine, γ-hydroxybutyric acid, N-acetyl-D-glucosamine, glycogen, itaconic acid, glycyl-L-glutamic acid, α-ketobutyricacid and putrescine, whereas it had received decreased capacities in using the other 20 carbon sources in Biolog Eco-plates. Redundancy analysis (RDA) revealed that the physico-chemical properties, indices of bacterial diversity, and C metabolic traits were positively correlated with the appearance of novel sequences under BC2 treatment. Our study indicates that the addition of biochar can increase culturable microbial abundance and shift bacterial genetic structure without enhancing their capacities in utilizing C sources in Biolog Eco-plates, which could be associated with the porous structure and nutrients from biochar.

Cow manure and cow manure-derived biochar application as a soil amendment for reducing cadmium availability and accumulation by Brassica chinensis L. in acidic red soil

Yasmin Khan Kiran, Ali Barkat, CUI Xiao-qiang, FENG Ying, PAN Feng-shan, TANG Lin, YANG Xiao-e

2017, 16(03): 725-734. DOI: 10.1016/S2095-3119(16)61488-0

Abstract ( ) PDF in ScienceDirect

Organic amendment is a promising, in situ phytostabilization approach to alleviate the phytotoxic effects of heavy metal contaminated soils. The aim of this study was to evaluate the feasibility of cow manure (CM) and its derived biochar (CMB) as a soil amendment on cadmium (Cd) availability and accumulation in low and high Cd-accumulating cultivars of Brassica chinensis L. grown in an acidic red soil. CM and CMB were applied to Cd-contaminated acidic red soil at the rates of 0, 3.0 and 6.0% (w/w). Application of CMB was significantly more effective than that of CM, as it reduced the availability of Cd in soil by 34.3–69.9% and its bioaccumulation in the low Cd accumulator, Aijiaoheiye 333, by 51.2 and 67.4%, respectively. The addition of CMB significantly increased the extractability and accumulation of trace metals (Zn, Mn, Fe, and Cu) by plants and improved plant biomass production. CMB application, combined with utilizing low Cd accumulating cultivars represents a new, sustainable strategy to alleviate the toxic effects on Cd and improve food safety.

Salicylic acid alleviates postharvest chilling injury of sponge gourd (Luffa cylindrica)

HAN Cong, ZUO Jin-hua, WANG Qing, DONG Hai-zhou, GAO Li-pu

2017, 16(03): 735-741. DOI: 10.1016/S2095-3119(16)61390-4

Abstract ( ) PDF in ScienceDirect

Effect of salicylic acid (SA) on chilling injury (CI) of sponge gourd during storage (9 days, 9°C) plus shelf life (2 days, 20°C) was evaluated in this study. SA treatment at the concentration of 1.5 mmol L^–1 significantly reduced postharvest CI of sponge gourds. Besides, the application of SA could effectively decrease the electrolyte leakage, reduce the accumulation of malondialdehyde (MDA) and total phenolics, enhance the activities of antioxidant enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), and inhibit the activities of phenylalanine ammonia-lyase (PAL) and polyphenol oxidase (PPO). The beneficial effects of SA could be attributed to preserved membrane integrity, inhibited membrane peroxidation, enhanced antioxidant system and suppressed activities of browning related enzymes. In a sense, SA as a postharvest tool may be commercially used in alleviating CI of sponge gourd.

Effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli O₈ and its cell surface characteristics

CHEN Yu-jie, WANG Chun-jie, HOU Wen-qian, WANG Xiao-shuo, GALI Bing-ga, HUASAI Si-mu-ji-de, YANG Si-qin, WU A-qi-ma, ZHAO Yu-fei, WU Ying-ga, CHEN Ao-ri-ge-le

2017, 16(03): 742-748. DOI: 10.1016/S2095-3119(16)61516-2

Abstract ( ) PDF in ScienceDirect

The effects of antibacterial compounds produced by Saccharomyces cerevisiae in Koumiss on pathogenic Escherichia coli O₈ and its cell surface characteristics were investigated. S. cerevisiae isolated from Koumiss produced antibacterial compounds which were active against pathogenic E. coli O₈ as determined by the Oxford cup method. The aqueous phases from S. cerevisiae at pH=2.0 (S2) and pH=8.0 (S8) were extracted and tested, respectively. The organic acids of S2 and S8 were determined by high performance liquid chromatography (HPLC), and the concentrations of killer toxins were determined by enhanced bicinchoninic acid (BCA) Protein Assay Kit. The minimum inhibition concentration (MIC) and the minimum bactericidal concentration (MBC) of S2 and S8 on E. coli O₈ were determined by the broth microdilution method. The effects of S2 and S8 on the growth curve of E. coli O₈ were determined by turbidimetry, and the hydrophobicities of E. coli O₈ cell surface were determined using the microbial adhesion to solvents method, the permeation of E. coli O₈ cell membrane were determined by the o-nitrophenyl-β-D-galactoside (ONPG) method. Aqueous phases at pH 2.0 and 8.0 had larger inhibition zones and then S2 and S8 were obtained by freeze-drying. The main component in S2 was citric acid and it was propanoic acid in S8. Other organic acids and killer toxins were also present. Both the MICs of S2 and S8 on E. coli O₈ were 0.025 g mL^–1, the MBCs were 0.100 and 0.200 g mL^–1, respectively. The normal growth curve of E. coli O₈ was S-shaped, however, it changed after addition of S2 and S8. E. coli O₈ was the basic character, and had a relatively hydrophilic surface. The hydrophobicity of E. coli O₈ cell surface and the permeation of E. coli O₈ cell membrane were increased after adding S2 and S8. The present study showed that S2 and S8 inhibit the growth of pathogenic E. coli O₈ and influence its cell surface characteristics.

Complete genome sequence of Bacillus thuringiensis Bt185, a potential soil insect biocontrol agent

LI Yan-qiu, SHU Chang-long, SHAN Yue-ming, GENG Li-li, SONG Fu-ping, ZHANG Jie

2017, 16(03): 749-751. DOI: 10.1016/S2095-3119(16)61422-3

Abstract ( ) PDF in ScienceDirect

Bacillus thuringiensis Bt185 and its insecticidal spectrum-expanded engineering strains are considered as potential biocontrol agents to soil insect Holotrichia parallela, Holotrichia oblita or Anomala corpulenta. Here we reported the complete genome of strain Bt185, it harbors eight plasmids, and plasmid pBT1850294 carries three cry8 genes.

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