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Hole fertilization in the root zone facilitates maize yield and nitrogen utilization by mitigating potential N loss and improving mineral N accumulation
SHI Wen-xuan, ZHANG Qian, LI Lan-tao, TAN Jin-fang, XIE Ruo-han, WANG Yi-lun
2023, 22 (4): 1184-1198.   DOI: 10.1016/j.jia.2022.09.018
Abstract248)      PDF in ScienceDirect      

Reducing environmental impacts and improving N utilization are critical to ensuring food security in China.  Although root-zone fertilization has been considered an effective strategy to improve nitrogen use efficiency (NUE), the effect of controlled-release urea (CRU) applied in conjunction with normal urea in this mode is unclear.  Therefore, a 3-year field experiment was conducted using a no-N-added as a control and two fertilization modes (FF, furrow fertilization by manual trenching, i.e., farmer fertilizer practice; HF: root-zone hole fertilization by point broadcast manually) at 210 kg N ha–1 (controlled-release:normal fertilizer=5:5), along with a 1-year in-situ microplot experiment.  Maize yield, NUE and N loss were investigated under different fertilization modes.  The results showed that compared with FF, HF improved the average yield and N recovery efficiency by 8.5 and 22.3% over three years, respectively.  HF had a greater potential for application than FF treatment, which led to increases in dry matter accumulation, total N uptake, SPAD value and LAI.  In addition, HF remarkably enhanced the accumulation of 15N derived from fertilizer by 17.2% compared with FF, which in turn reduced the potential loss of 15N by 43.8%.  HF increased the accumulation of N in the tillage layer of soils at harvest for potential use in the subsequent season relative to FF.  Hence, HF could match the N requirement of summer maize, sustain yield, improve NUE and reduce environmental N loss simultaneously.  Overall, root-zone hole fertilization with blended CRU and normal urea can represent an effective and promising practice to achieve environmental integrity and food security on the North China Plain, which deserves further application and investigation.

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Isolation and functional analysis of SrMYB1, a direct transcriptional repressor of SrUGT76G1 in Stevia rebaudiana

ZHANG Ting, ZHANG Yong-xia, SUN Yu-ming, XU Xiao-yang, WANG Yin-jie, CHONG Xin-ran, YANG Yong-heng and YUAN Hai-yan
2023, 22 (4): 1058-1067.   DOI: 10.1016/j.jia.2023.03.001
Abstract212)      PDF in ScienceDirect      

SrUGT76G1, the most well-studied diterpene glycosyltransferase in Stevia rebaudiana, is key to the biosynthesis of economically important steviol glycosides (SGs).  However, the molecular regulatory mechanism of SrUGT76G1 has rarely been explored.  In this study, we identified a MYB transcription factor, SrMYB1, using a yeast one-hybrid screening assay.  SrMYB1 belongs to the typical R2R3-type MYB protein and is specifically localized in the nucleus with strong transactivation activity.  The transcript of SrMYB1 is predominantly accumulated in flowers, but is also present at a lower level in leaves.  Yeast one-hybrid and electrophoretic mobility shift assays verified that SrMYB1 binds directly to the MYB binding sites in the F4-3 fragment (+50–(–141)) of the SrUGT76G1 promoter.  Furthermore, we found that SrMYB1 could significantly repress the expression of SrUGT76G1 in both epidermal cells of tobacco leaves and stevia callus.  Taken together, our results demonstrate that SrMYB1 is an essential upstream regulator of SrUGT76G1 and provide novel insight into the regulatory network for the SGs metabolic pathway in S. rebaudiana.

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Functional analysis of MdSUT2.1, a plasma membrane sucrose transporter from apple
ZHANG Bo, FAN Wen-min, ZHU Zhen-zhen, WANG Ying, ZHAO Zheng-yang
2023, 22 (3): 762-775.   DOI: 10.1016/j.jia.2022.09.012
Abstract267)      PDF in ScienceDirect      

Sugar content is a determinant of apple (Malus×domestica Borkh.) sweetness.  However, the molecular mechanism underlying sucrose accumulation in apple fruit remains elusive.  Herein, this study reported the role of the sucrose transporter MdSUT2.1 in the regulation of sucrose accumulation in apples.  The MdSUT2.1 gene encoded a protein with 612 amino acid residues that could be localized at the plasma membrane when expressed in tobacco leaf protoplasts.  MdSUT2.1 was highly expressed in fruit and was positively correlated with sucrose accumulation during apple fruit development.  Moreover, complementary growth assays in a yeast mutant validated the sucrose transport activity of MdSUT2.1.  MdSUT2.1 overexpression in apples and tomatoes resulted in significant increases in sucrose, fructose, and glucose contents compared to the wild type (WT).  Further analysis revealed that the expression levels of sugar metabolism- and transport-related genes SUSYs, NINVs, FRKs, HXKs, and TSTs increased in apples and tomatoes with MdSUT2.1 overexpression compared to WT.  Finally, unlike the tonoplast sugar transporters MdTST1 and MdTST2, the promoter of MdSUT2.1 was not induced by exogenous sugars.  These findings provide valuable insights into the molecular mechanism underlying sugar accumulation in apples.

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Characterization of a blaCTX-M-3, blaKPC-2 and blaTEM-1B co-producing IncN plasmid in Escherichia coli of chicken origin

WANG Wen-jing, WANG Yi-fu, JIN Ya-jie, SONG Wu-qiang, LIN Jia-meng, ZHANG Yan, TONG Xin-ru, TU Jian, LI Rui-chao, LI Tao
2023, 22 (1): 320-324.   DOI: 10.1016/j.jia.2022.08.075
Abstract338)      PDF in ScienceDirect      

An extensively drug-resistant (XDR) Escherichia coli strain 258E was isolated from an anal swab sample of a chicken farm of Anhui province in China. Genomic analyses indicated that the strain 258E harbors an incompatibility group N (IncN) plasmid pEC258-3, which co-produces blaCTX-M-3, blaKPC-2, blaTEM-1B, qnrS1, aac(6')-Ib-cr, dfrA14, arr-3, and aac(6')-Ib3. Multiple genome arrangement analyses indicated that pEC258-3 is highly homologous with pCRKP-1-KPC discovered in Klebsiella pneumoniae from a patient. Furthermore, conjugation experiments proved that plasmid pEC258-3 can be transferred horizontally and may pose a significant potential threat in animals, community and hospital settings.

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Allele mining of wheat ABA receptor at TaPYL4 suggests neo-functionalization among the wheat homoeologs
WU Bang-bang, SHI Meng-meng, Mohammad POURKHEIRANDISH, ZHAO Qi, WANG Ying, YANG Chen-kang, QIAO Ling, ZHAO Jia-jia, YAN Su-xian, ZHENG Xing-wei, ZHENG Jun
2022, 21 (8): 2183-2196.   DOI: 10.1016/S2095-3119(21)63699-7
Abstract213)      PDF in ScienceDirect      
ABA receptors (PYR/PYL/RCAR) play a central role in the water loss control of plants.  A previous report indicated that TaPYL4 is a critical gene in wheat that improves grain production under drought conditions and increases water use efficiency.  In this study, we analyzed the sequence polymorphisms and genetic effects of TaPYL4s.  Based on isolated TaPYL4 genes from chromosomes 2A, 2B and 2D, three haplotypes were detected in the promoter region of TaPYL4-2A, and two haplotypes were present in TaPYL4-2B and TaPYL4-2D, respectively.  Marker/trait association analysis indicated that TaPYL4-2A was significantly associated with plant height in 262 Chinese wheat core collection accessions, as well as the drought tolerance coefficient (DTC) for plant height in 239 wheat varieties from Shanxi Province in multiple environments.  However, the frequencies of favored drought-tolerant haplotype TaPYL4-2A-Hap2 were considerably low, accounting for only 10%, and lines with this certain Hap could be reserved in the breeding program.  TaPYL4-2B was significantly associated with grain number, and the favored haplotype TaPYL4-2B-Hap1 was the dominant allele of above 90% in the collection.  For TaPYL4-2D, there were no significant differences in these traits between the two haplotypes in either of the two panels.  These results indicate that variation might lead to functional differentiation among the homoeologs and the haplotypes had undergone artificial selection during breeding.  Two molecular markers developed to distinguish these haplotypes could be used for breeding in water-limited regions.

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Kiwifruit (Actinidia chinensis ‘Hongyang’) cytosolic ascorbate peroxidases (AcAPX1 and AcAPX2) enhance salinity tolerance in Arabidopsis thaliana
GUO Xiu-hong, HE Yan, ZHANG Yu, WANG Yi, HUANG Sheng-xiong, LIU Yong-sheng, LI Wei
2022, 21 (4): 1058-1070.   DOI: 10.1016/S2095-3119(21)63652-3
Abstract144)      PDF in ScienceDirect      
Ascorbate peroxidase (APX) plays a key role in scavenging reactive oxygen species (ROS) in higher plants.  However, there is very little information available on the APXs in kiwifruit (Actinidia), which is an economically and nutritionally important horticultural crop with exceptionally high ascorbic acid (AsA) accumulation.  This study aims to identify and characterize two cytosolic APX genes (AcAPX1 and AcAPX2) derived from A. chinensis ‘Hongyang’.  The constitutive expression pattern was determined for both AcAPX1 and AcAPX2, and showed relatively higher expression abundances of AcAPX1 in leaf and AcAPX2 in root.  Transcript levels of AcAPX1 and AcAPX2 were increased in kiwifruit roots treated with NaCl.  Subcellular localization assays using GFP-fusion proteins in Arabidopsis protoplasts showed that both AcAPX1 and AcAPX2 are targeted to the cytosol.  Recombinant AcAPX1 or AcAPX2 proteins were successfully expressed in the prokaryotic expression system and their individual ascorbate peroxidase activities were determined.  Finally, constitutive over-expression of AcAPX1 or AcAPX2 could dramatically increase total AsA, glutathione level and salinity tolerance under NaCl stress in Arabidopsis thaliana.  Our findings revealed that cytosolic AcAPX1/2 may play an important protective role in the responses to unfavorable environmental stimuli in kiwifruit.
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Incidence and prevalence levels of three aphid-transmitted viruses in crucifer crops in China
ZHANG Xiao-yan, PENG Yan-mei, XIANG Hai-ying, WANG Ying, LI Da-wei, YU Jia-lin, HAN Cheng-gui
2022, 21 (3): 774-780.   DOI: 10.1016/S2095-3119(21)63618-3
Abstract235)      PDF in ScienceDirect      
Poleroviruses, which are distributed worldwide, infect many crops of economic importance and cause severe plant diseases.  Brassica yellows virus (BrYV), which has three genotypes, A, B, and C, is a newly identified polerovirus infecting crucifer crops in China, but its distribution is still unclear.  Here, we report the distribution and prevalence levels of the three BrYV genotypes in crucifer crops in China.  A total of 570 crucifer leaf samples randomly collected from 22 provinces, four ethnic minority autonomous regions, and three municipalities in China were tested for BrYV.  RT-PCR detection showed that 97 of the field samples were positive for BrYV, and the average incidence of BrYV was 17.0%.  The virus was detected in 22 provinces, with high incidences in north, northwest, and northeast China.  The multiplex RT-PCR amplification of the three BrYV genotypes revealed that both single and mixed infections occurred.  Among the BrYV infections, 38.1% were mix-infected by more than two viral genotypes, and 8.2% samples were mix-infected by three viral genotypes.  Our findings indicated a widespread prevalence of BrYV in China, and BrYV mixed infections with Turnip mosaic virus and Cucumber mosaic virus in crucifer crops are common.  This study is the first large-scale survey of BrYV in crucifer crops in China.  The information generated in this investigation will contribute to the national prevention and control of viral diseases.
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Chinese herbal formulae defend against Mycoplasma gallisepticum infection
WANG Ying-jie, LIANG Ya-xi, HU Fu-li, SUN Ying-fei, ZOU Meng-yun, LUO Rong-long, PENG Xiu-li
2022, 21 (10): 3026-3036.   DOI: 10.1016/j.jia.2022.07.038
Abstract321)      PDF in ScienceDirect      

Mycoplasma gallisepticum HS strain (MG-HS) is a pathogen that causes chronic respiratory disease (CRD) in chicken, which is characterized by host respiratory inflammatory damage, brings huge economic losses to the poultry industry.  Recently, emerging Chinese herbal medicines (CHM) have been used to treat CRD.  This study was aimed to investigate the preventive and therapeutic effects and their potential mechanisms of Chinese herbal medicinal formulae (CHMF), which consisted of 10 kinds of Chinese herbal medicine including Scutellaria, Houttuynia cordate and licorice, on MG-induced CRD in chickens.  With respect to the preventive effect, the results showed that CHMF could effectively recover the MG-induced decrease on body weight and feed conversion ratio.  Histopathological analysis showed that both prevention and treatment of CHMF could significantly alleviate the severe respiratory inflammation induced by MG infection.  Moreover, compared with the MG infection group, both the prevention groups and the treatment groups of CHMF could effectively reduce the expression of MG adhesion protein (pMGA1.2) to inhibit the proliferation of MG, and thus effectively inhibit the expression of MG-induced inflammatory factors interleukin-1β (IL-1β) and tumour necrosis factor-α (TNF-α).  In summary, these findings confirm that CHMF can protect chickens from various tissue damage caused by MG infection and has no adverse effects on the performance of chickens in the short term.  And its efficacy against MG is equal to or better than that of tiamulin.

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New geographic distribution and molecular diversity of Citrus chlorotic dwarf-associated virus in China
YANG Zhen, ZHANG Lan, ZHAO Jin-fa, ZHANG Xing-kai, WANG Ying, LI Tai-sheng, ZHANG Wei, ZHOU Yan
2022, 21 (1): 293-298.   DOI: 10.1016/S2095-3119(20)63601-2
Abstract125)      PDF in ScienceDirect      
In 2009, an emerging citrus viral disease caused by Citrus chlorotic dwarf-associated virus (CCDaV) was discovered in Yunnan Province of China.  However, the occurrence and spread of CCDaV in other citrus-growing provinces in China is unknown to date.  To better understand the distribution and molecular diversity of CCDaV in China, a total of 1 772 citrus samples were collected from 11 major citrus-growing provinces and were tested for CCDaV by PCR.  Among these, 134 citrus samples from Guangxi, Yunnan and Guangdong were tested positive for CCDaV, demonstrating that the occurrence and spread of CCDaV are increasing in China.  The complete genome sequences of 17 CCDaV isolates from different provinces and hosts were sequenced.  Comparisons of the whole-genome sequences of the 17 CCDaV isolates as well as the 15 isolates available in GenBank revealed that the sequence identity was about 99–100%, showing that the CCDaV isolates were highly conserved.  Phylogenetic studies showed that the 32 CCDaV isolates belonged to four different groups based on geographical origins and host species, and that CCDaV isolates from China and Turkey were clustered into different groups.  The results provide important information for clarifying the distribution and genetic diversity of CCDaV in China.
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Effects of shading stress during the reproductive stages on photosynthetic physiology and yield characteristics of peanut (Arachis hypogaea Linn.)
WANG Yi-bo, HUANG Rui-dong, ZHOU Yu-fei
2021, 20 (5): 1250-1265.   DOI: 10.1016/S2095-3119(20)63442-6
Abstract160)      PDF in ScienceDirect      
In intercropping systems, high-positioned crops often exert shading stress on low-positioned crops, thus affecting the agronomic characteristics of the low-positioned crops.  This study determined the mechanisms of photosynthetic, physiological and yield variations among peanut cultivars under shading stress.  Four peanut cultivars, S60, C4, P12, and YS151, were grown in the field and subjected to shading stress for 77 days during reproductive stages.  S60 and P12 showed lower yield and reduced biomass accumulation than C4 and YS151 under shading stress.  Shading stress induced higher antioxidant enzyme activities in S60 and P12, relative to C4 and YS151.  Under shading stress, S60 and P12 showed a higher light-trapping capability than C4 and YS151, which was associated with changes in chlorophyll (Chl) a and b contents, and Chl a/b ratio.  The net photosynthetic rate, stomatal conductance and transpiration rates of C4 and YS151 were decreased, but the intercellular CO2 concentration increased under shading stress.  The results demonstrated that non-stomatal limiting factors decreased the photosynthetic capacity of peanut under shading stress.  The maximum photochemical efficiency of PSII (Fv/Fm) and non-photochemical quenching (NPQ) were higher in S60 and P12 than in C4 and YS151 under shading stress.  These results suggest that S60 and P12 could absorb more light energy from weak light environments for photosynthesis than C4 and YS151 and dissipate the excess energy in the form of heat to improve their light protection ability.  This study explains the inter-variety differences in shading stress tolerance in peanut and provides physiological parameters for guiding the selection of shade-tolerant cultivars.
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Divergent responses of tiller and grain yield to fertilization and fallow precipitation: Insights from a 28-year long-term experiment in a semiarid winter wheat system
WANG Rui, WANG Ying, HU Ya-xian, DANG Ting-hui, GUO Sheng-li
2021, 20 (11): 3003-3011.   DOI: 10.1016/S2095-3119(20)63296-8
Abstract156)      PDF in ScienceDirect      
Tillering is an important phenological stage, which is strongly related to the yield in spike components and final grain yield during winter wheat growth.  Precipitation during the fallow season (fallow precipitation) influences tillering in winter wheat on the semi-arid Chinese Loess Plateau.  However, little work has been done regarding tiller number changes under various types of fertilization and amounts of fallow precipitation on a long-term scale.  Effects of fallow precipitation and fertilization on tiller were investigated in a winter wheat (Triticum aestivum L.) system in a 28-year field study (1990 to 2017) in a semiarid agro-ecosystem.  Tiller number, spike number and grain yield were measured in four fertilization conditions: control without fertilizer (CK); mineral nitrogen fertilizer alone (N); mineral phosphorus fertilizer alone (P); mineral nitrogen and phosphorus fertilizer together (NP).  Based on the long-term annual fallow precipitation, dry years (<mean annual fallow precipitation) and wet years (>mean annual fallow precipitation) were distinguished.  Phosphorus fertilization alone significantly increased the mean annual tiller number (23%), and the increase in tiller number was higher in wet years (29%) than in the dry years (17%).  However, nitrogen fertilization alone had little effect on mean tiller number, while nitrogen and phosphorus together significantly increased mean annual tiller number (30%), mean tiller number in wet years (45%) and mean tiller number in dry years (17%).  Tiller number was significantly and positively correlated with fallow precipitation in dry years for all fertilizer treatments, whereas it was weakly and either positively or negatively correlated with fallow precipitation in wet years depending on the treatment.  This study found positive correlations between tiller number and fallow precipitation in the CK and NP treatments, and it found negative correlations between tiller number and fallow precipitation in the treatments with nitrogen fertilization alone or phosphorous fertilization alone in wet years.  Understanding the impacts of fallow precipitation and fertilization on tiller development shed light on ways to improve crop production in rain-fed agricultural regions.
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A case-based method of selecting covariates for digital soil mapping
LIANG Peng, QIN Cheng-zhi, ZHU A-xing, HOU Zhi-wei, FAN Nai-qing, WANG Yi-jie
2020, 19 (8): 2127-2136.   DOI: 10.1016/S2095-3119(19)62857-1
Abstract116)      PDF in ScienceDirect      
Selecting a proper set of covariates is one of the most important factors that influence the accuracy of digital soil mapping (DSM).  The statistical or machine learning methods for selecting DSM covariates are not available for those situations with limited samples.  To solve the problem, this paper proposed a case-based method which could formalize the covariate selection knowledge contained in practical DSM applications.  The proposed method trained Random Forest (RF) classifiers with DSM cases extracted from the practical DSM applications and then used the trained classifiers to determine whether each one potential covariate should be used in a new DSM application.  In this study, we took topographic covariates as examples of covariates and extracted 191 DSM cases from 56 peer-reviewed journal articles to evaluate the performance of the proposed case-based method by Leave-One-Out cross validation.  Compared with a novices’ commonly-used way of selecting DSM covariates, the proposed case-based method improved more than 30% accuracy according to three quantitative evaluation indices (i.e., recall, precision, and F1-score).  The proposed method could be also applied to selecting the proper set of covariates for other similar geographical modeling domains, such as landslide susceptibility mapping, and species distribution modeling.
 
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Alphaherpesvirus-vectored vaccines against animal diseases: Current progress
HU Yang, WANG Ming-shu, CHENG An-chun, JIA Ren-yong, YANG Qiao, WU Ying, LIU Ma-feng, ZHAO Xin-xin, ZHU De-kang, CHEN Shun, ZHANG Sha-qiu, WANG Yin, GAO Qun, OU Xu-min, MAO Sai, WEN Xing-jian, XU Zhi-wen, CHEN Zheng-li, ZHU Ling, LUO Qi-hui, TIAN Bin, PAN Lei-chang, Mujeeb Ur REHMAN, LIU Yun-ya, YU Yan-ling, ZHANG Ling, CHEN Xiao-yue
2020, 19 (8): 1928-1940.   DOI: 10.1016/S2095-3119(20)63175-6
Abstract242)      PDF in ScienceDirect      
Recombinant virus-vectored vaccines are novel agents that can effectively activate specific and nonspecific immunity, are multivalent and multieffective, and have high safety ratings.  Animal alphaherpesviruses have a large genome, contain multiple nonessential regions that do not affect viral replication and are capable of accepting the insertion of an exogenous gene and expressing the antigen protein.  Furthermore, animal alphaherpesviruses have a wide host spectrum, can replicate in the host and continuously stimulate the animal to produce immunity to the corresponding pathogen, thus making them ideal carriers for recombinant virus-vectored vaccines.  With the development of gene-editing technology, recombinant viruses capable of expressing foreign genes can be constructed by various methods.  Currently, studies on recombinant virus-vectored vaccines constructed based on animal alphaherpesviruses have involved poultry, pigs, cattle, sheep, and companion animals.  Studies have shown that the construction of recombinant animal alphaherpesviruses enables the acquisition of immunity to multiple diseases.  This article mainly summarizes the current progress on animal alphaherpesvirus-vectored vaccines, aiming to provide reference for the development of new animal alphaherpesvirus-vectored vaccines.
 
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Differentially expressed miRNAs in anthers may contribute to the fertility of a novel Brassica napus genic male sterile line CN12A
Dong Yun, Wang Yi, Jin Feng-wei, Xing Li-juan, Fang Yan, Zhang Zheng-ying, ZOU Jun-jie, Wang Lei, Xu Miao-yun
2020, 19 (7): 1731-1742.   DOI: 10.1016/S2095-3119(19)62780-2
Abstract95)      PDF in ScienceDirect      
In Brassica napus L. (rapeseed), complete genic male sterility (GMS) plays an important role in the utilization of heterosis.  Although microRNAs (miRNAs) play essential regulatory roles during bud development, knowledge of how GMS is regulated by miRNAs in rapeseed is rather limited.  In this study, we obtained a novel recessive GMS system, CN12AB.  The sterile line CN12A has defects in tapetal differentiation and degradation.  Illumina sequencing was employed to examine the expression of miRNAs in the buds of CN12A and the fertile line CN12B.  We identified 85 known miRNAs and 120 novel miRNAs that were expressed during rapeseed anther development.  When comparing the expression levels of miRNAs between CN12A and CN12B, 19 and 18 known miRNAs were found to be differentially expressed in 0.5–1.0 mm buds and in 2.5–3.0 mm buds, respectively.  Among these, the expression levels of 14 miRNAs were higher and the levels of 23 miRNAs were lower in CN12A compared with CN12B.  The predicted target genes of these differentially expressed miRNAs encode protein kinases, F-box domain-containing proteins, MADS-box family proteins, SBP-box gene family members, HD-ZIP proteins, floral homeotic protein APETALA 2 (AP2), and nuclear factor Y, subunit A.  These targets have previously been reported to be involved in pollen development and male sterility, suggesting that miRNAs might act as regulators of GMS in rapeseed anthers.  Furthermore, RT-qPCR data suggest that one of the differentially expressed miRNAs, bna-miR159, plays a role in tapetal differentiation by regulating the expression of transcription factor BnMYB101 and participates in tapetal degradation and influences callose degradation by manipulating the expression of BnA6.  These findings contribute to our understanding of the roles of miRNAs during anther development and the occurrence of GMS in rapeseed.
 
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Genome-wide identification and expression analysis of StPP2C gene family in response to multiple stresses in potato (Solanum tuberosum L.)
WANG Yi-fan, LIAO Yu-qiu, WANG Ya-peng, YANG Jiang-wei, ZHANG Ning, SI Huai-jun
2020, 19 (6): 1609-1624.   DOI: 10.1016/S2095-3119(20)63181-1
Abstract153)      PDF in ScienceDirect      
The plant protein phosphatase 2Cs (PP2Cs) play an essential role in response to stress and abscisic acid (ABA) signaling pathway.  However, to date, no systemic characterization of the PP2Cs has yet been conducted in potato (Solanum tuberosum L.).  In the study, a comprehensive research was performed on genome-wide identification and expression analysis of StPP2C genes in potato.  A total of 78 potato StPP2C genes were identified based on specific structure of PP2C domain, which were distributed across 11 out of 12 potato chromosomes and divided into 12 (A–L) phylogenetic branches.  The result from gene duplication analysis showed that 14 StPP2Cs were involved in gene tandem duplication and 8 genes formed fragment duplication events, which indicated that both tandem and fragment duplication contributed to the expansion of the gene family in evolution.  Exon–intron structural analysis showed that they had a wide range of exon numbers.  Analysis of protein conservative motif demonstrated that StPP2Cs contained more similar motif structures in the same phylogenetic branches.  The cis-elements in StPP2C gene promoter regions were mainly responded to light, phytohormone and abiotic stress.  Most of them exhibited tissue-specific expression patterns, and some members could differentially express under abiotic stress.  The evidence suggested that StPP2C genes may contribute to different functions in several physiological stress and environmental stress conditions.  This study could provide new insights to further investigate StPP2C functional characteristics responding to various stresses in potato.
 
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Feeding effects of dsNPF interference in Ostrinia furnacalis
CUI Hong-ying, WANG Yuan, PENG Xin, WANG Yi-tong, ZHAO Zhang-wu
2020, 19 (6): 1475-1481.   DOI: 10.1016/S2095-3119(19)62788-7
Abstract96)      PDF in ScienceDirect      
The corn borer is a world-wide agricultural pest.  In this study, a high-efficiency RNAi method was explored to knock down the neuropeptide F (NPF) to determine if NPF regulates larval feeding in the Asian corn borer, Ostrinia furnacalis.  Results showed that the expression peaks of npf mRNA in both midgut and fat body are at 28 h of early 5th instar larvae.  When NPF was knocked down either by feeding dsNPF-containing artificial diet or direct dsNPF injection into the 5th instar larvae, npf expression was effectively inhibited in the midgut.  Larval feeding, body weight and development time were significantly impacted.  In contrast, the expression of the npf receptors npfr1 and npfr2 in fat body of 5th instar larvae was significantly increased by dsNPF injection.  These results indicate that NPF can be effectively knocked down in O. furnacalis, and dsNPF by injection is an effective and fast way to silence npf expression.  This study provides a critical basis for further exploration in mechanism of feeding regulation in O. furnacalis.
 
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OsHemA gene, encoding glutamyl-tRNA reductase (GluTR) is essential for chlorophyll biosynthesis in rice (Oryza sativa)
ZENG Zhao-qiong, LIN Tian-zi, ZHAO Jie-yu, ZHENG Tian-hui, XU Le-feng, WANG Yi-hua, LIU Ling-long, JIANG Ling, CHEN Sai-hua, WAN Jian-min
2020, 19 (3): 612-623.   DOI: 10.1016/S2095-3119(19)62710-3
Abstract142)      PDF in ScienceDirect      
Chlorophyll (Chl) biosynthesis is essential for photosynthesis and plant growth.  Glutamyl-tRNA reductase (GluTR) catalyzes glutamyl-tRNA into glutamate-1-semialdehyde (GSA) and initiates the chlorophyll biosynthesis.  Even though the main role of GluTR has been established, the effects caused by natural variations in its corresponding gene remain largely unknown.  Here, we characterized a spontaneous mutant in paddy field with Chl biosynthesis deficiency, designated as cbd1.  With intact thylakoid lamellar structure, the cbd1 plant showed light green leaves and reduced Chl and carotenoids (Cars) content significantly compared to the wild type.  By map-based gene cloning, the mutation was restricted within a 57-kb region on chromosome 10, in which an mPingA miniature inverted-repeat transposable element (MITE) inserted in the promoter region of OsHemA gene.  Both leaf color and the pigment contents in cbd1 were recovered in a complementation test, confirming OsHemA was responsible for the mutant phenotype.  OsHemA was uniquely predicted to encode GluTR and its expression level was dramatically repressed in cbd1.  Transient transformation in protoplasts demonstrated that GluTR localized in chloroplasts and a signal peptide exists in its N-terminus.  A majority of Chl biosynthesis genes, except for POR and CHLG, were down-regulated synchronously by the repression of OsHemA, suggesting that an attenuation occurred in the Chl biosynthesis pathway.  Interestingly, we found major agronomic traits involved in rice yield were statistically unaffected, except for the number of full grains per panicle was increased in cbd1.  Collectively, OsHemA plays an essential role in Chl biosynthesis in rice and its weak allele can adjust leaf color and Chls content without compromise to rice yield.
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The expression of Lin28B was co-regulated by H3K4me2 and Wnt5a/β-catenin/TCF7L2
ZHANG Ya-ni, HU Cai, WANG Ying-jie, ZUO Qi-sheng, LI Bi-chun
2020, 19 (12): 3054-3064.   DOI: 10.1016/S2095-3119(20)63441-4
Abstract121)      PDF in ScienceDirect      
Lin28A and Lin28B are homologous RNA-binding proteins that participate in the development of primordial germ cells.  The mechanisms underlying expression and regulation of Lin28A have been well documented, but such information for Lin28B is limited.  In this study, a fragment of the Lin28B promoter was cloned, the pEGFP-pLin28B vector was constructed.  DF-1 chicken fibroblasts were transfected and the expression of green fluorescent protein (GFP) was measured.  Furtherly, Lin28B promoter of different lengths fragments was cloned using the chromosome-walking method and the fragments were ligated into the PGL3-Basic vector, and transfected into DF-1 cells.  Results of dual-luciferase reporter assay showed that the core of the Lin28B promoter was included in the sequence from –1 431 to –1 034 bp.  The binding sites of the transcription factor TCF7L2 was showed within this sequence by bioinformatics analysis.  The promoter activity of Lin28B was downregulated (P<0.05) when the TCF7L2 binding site was mutated.  Further experiments suggested that Lin28B promoter activity responded to the activation or inhibition of Wnt signaling.  Results of chromatin immunoprecipitation and quantitative PCR showed that β-catenin-TCF7L2 may be enriched in the Lin28B promoter core area.  In vivo and in vitro activation or inhibition of Wnt signaling significantly up- or down-regulated (P<0.05) Lin28B expression.  H3K4me2 enriched in the promoter of Lin28B, which affected the regulation of Wnt signaling to Lin28B.  In conclusion, our results showed that H3K4me2 and Wnt5a/β-catenin/TCF7L2 were the positive regulators of Lin28B expression.  Findings of this study may lay a theoretical foundation for illuminating the mechanism underlying Lin28B expression.
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Crop diversity and pest management in sustainable agriculture
HE Han-ming, LIU Li-na, Shahzad Munir, Nawaz Haider Bashir, WANG Yi, YANG Jing, LI Cheng-yun
2019, 18 (9): 1945-1952.   DOI: 10.1016/S2095-3119(19)62689-4
Abstract195)      PDF in ScienceDirect      
Large-scale crop monocultures facilitate the proliferation and increasing prevalence of diseases and pest insects.  Many studies highlight the impacts of plant diversification upon pathogens, and the population dynamics of insects and beneficial organism in agricultural ecosystems.  These studies provide evidence that habitat manipulation techniques such as intercropping, relay, and rotation can significantly improve disease and pest management.  This review introduces the concept of crop diversity, considers recent insights and mechanisms underlying crop diversity, and discusses its potential for improving sustainable agricultural practices.  Recently, the phytobiomes resulting from increased crop diversity are increasingly recognized for their contribution to disease and pest control.  Further, understanding the interactions between pathogens or pests with their host phytobiome may lead to novel options for the prevention of pests.  Recent advances in the agricultural systems include: (i) a better understanding of the mechanisms of interactions between crop species and genotypes; (ii) ecological progress including a better understanding of the context-dependency of those interactions; and (iii) the role of microtopographic variation in agricultural systems for priming basal resistance to multiple pests and pathogens by intercropped crops.  We also highlight recent progress in China and the potential options for habitat management and design that enhance the ecological role of biodiversity in agroecosystems.
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Phosphorylation of sarcoplasmic and myofibrillar proteins in three ovine muscles during postmortem ageing
WANG Ying, LI Xin, LI Zheng, DU Man-ting, ZHU Jie, ZHANG She-qi, ZHANG De-quan
2019, 18 (7): 1643-1651.   DOI: 10.1016/S2095-3119(19)62653-5
Abstract308)      PDF in ScienceDirect      
This study aimed to examine changes in phosphorylation of sarcoplasmic and myofibrillar proteins from longissimus lumborum, semitendinosus, and psoas major muscles during postmortem ageing for 5 d.  These sarcoplasmic and myofibrillar proteins were separated using sodium dodecyl sulfate-polyacrylamide gel electrophoresis and stained with phosphorous and protein specific stains.  Myofibril fragmentation index, pH, the content of lactic acid and the relative activity of μ-calpain in three ovine muscles were measured.  These results showed that the relative phosphorylation level of sarcoplasmic and myofibrillar proteins of psoas major muscle were lower compared with longissimus lumborum and semitendinosus muscles (P<0.05).  The pH of psoas major muscle was the lowest at 0.5 h postmortem, and the highest after 12 h postmortem (P<0.05).  In addition, the relative activity of μ-calpain was higher within 5 d postmortem and myofibril fragmentation index was higher after 1 d postmortem in psoas major muscle than those of longissimus lumborum and semitendinosus muscles (P<0.05).  The sarcoplasmic protein phosphorylation may regulate the rate of pH decline to influence the μ-calpain activity and then proteolysis of proteins consequently.  This study gives a new perspective of the mechanism of postmortem meat tenderization.
 
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The autophagy gene ATG8 affects morphogenesis and oxidative stress tolerance in Sporisorium scitamineum
ZHANG Bin, CUI Guo-bing, CHANG Chang-qing, WANG Yi-xu, ZHANG Hao-yang, CHEN Bao-shan, DENG Yi-zhen, JIANG Zi-de
2019, 18 (5): 1024-1034.   DOI: 10.1016/S2095-3119(18)62109-4
Abstract232)      PDF in ScienceDirect      
The basidiomycetous fungus Sporisorium scitamineum causes sugarcane smut that leads to severe economic losses in the major sugarcane growing areas in China, India and Brazil, etc.  Autophagy is a conserved pathway in eukaryotes for bulk degradation and cellular recycling, and was shown to be important for fungal cell growth, development, and pathogenicity.  However, physiological function of autophagy has not been studied in S. scitamineum.  In this study, we identified a conserved Atg8 protein, named as SsAtg8 and characterized its function. Our results showed that autophagy was blocked in the ssatg8Δ mutant, in nitrogen starvation.  The ssatg8Δ mutant formed pseudohypha frequently and was hypersensitive to oxidative stress.  However, mating or filamenation was unaffected in the ssatg8Δ mutant in vitro.  Overall we demonstrate that autophagy is dispensable for S. scitamineum mating/filamentation, while critical for oxidative stress tolerance and proper morphology in sporidial stage.   
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Invasion biology of the cotton mealybug, Phenacoccus solenopsis Tinsley: Current knowledge and future directions
TONG Hao-jie, AO Yan, LI Zi-hao, WANG Ying, JIANG Ming-xing
2019, 18 (4): 758-770.   DOI: 10.1016/S2095-3119(18)61972-0
Abstract307)      PDF (825KB)(667)      
The cotton mealybug Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae), which is native to North America, emerged as a major invasive insect pest of multiple crops in Asia at the beginning of the 21st century.  Considering the economic significance of this insect and its rapid worldwide spread, we examined the possible factors driving its invasions.  In this paper, we summarize the life history traits of P. solenopsis conceivably related to population development in invaded regions; these traits include its use of diverse host plants, reproductive capacity and mode, adaptation to temperature, response to food shortage, and insecticidal resistance.  Then, focusing on the multiple trophic interactions that may promote or hinder invasion, we review the mutualistic relationship of this mealybug with ants and predation and parasitism by natural enemies.  Finally, we suggest topics for future research and provide our perspective on the biological invasions of this mealybug.  We speculate that specific biological factors associated with this mealybug, particularly its wide host range, high reproductive potential, evolved changes in life history traits, and mutualism with ants have played important roles in its invasions, allowing this pest to become established and rapidly increase its population upon its introduction into new regions.
 
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A dCAPS marker developed from a stress associated protein gene TaSAP7-B governing grain size and plant height in wheat
WANG Yi-xue, XU Qiao-fang, CHANG Xiao-ping, HAO Chen-yang, LI Run-zhi, JING Rui-lian
2018, 17 (2): 276-284.   DOI: 10.1016/S2095-3119(17)61685-X
Abstract755)      PDF in ScienceDirect      
Stress associated proteins (SAPs) are the A20/AN1 zinc-finger proteins which confer to abiotic stresses in plants.  In this study, TaSAP7-B, including two AN1 domains, was isolated from B genome of wheat (Triticum aestivum L.).  Sequencing analysis on TaSAP7-B illustrated one InDel (insertion-deletion) and one SNP (single nucleotide polymorphism) in the promoter region while no diversity was observed in the coding region.  On the basis of SNP in the promoter region (–260 bp), a dCAPS (derived cleaved amplified polymorphic sequences) marker SNP-260 was developed for TaSAP7-B.  Using a natural population consisting of 262 wheat accessions, significant associations were detected between the marker SNP-260 and agronomic traits, such as plant height (PH), peduncle length (PL), length of penultimate internode (LPI), number of spike per plant (NSP), and 1 000-grain weight (TGW).  Two genotypes were identified using marker SNP-260 in the natural population.  Among them, the genotypes possessing C allele exhibited a higher TGW and shorter PH than the T genotypes.  Hence, base C was considered as the superior allele.  The dCAPS marker of TaSAP7-B can be instrumental for marker-assisted selection for high grain size and short plant height.  
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Maize ABP2 enhances tolerance to drought and salt stress in transgenic Arabidopsis
ZONG Na, LI Xing-juan, WANG Lei, WANG Ying, WEN Hong-tao, LI Ling, ZHANG Xia, FAN Yun-liu, ZHAO Jun
2018, 17 (11): 2379-2393.   DOI: 10.1016/S2095-3119(18)61947-1
Abstract375)      PDF (35768KB)(362)      
Abiotic stresses, especially drought and salt, severely affect maize production, which is one of the most important cereal crops in the world.  Breeding stress-tolerant maize through biotechnology is urgently needed to maintain maize production.  Therefore, it is important to identify new genes that can enhance both drought and salt stress tolerance for molecular breeding. In this study, we identified a maize ABA (abscisic acid)-responsive element (ABRE) binding protein from a 17-day post-pollination (dpp) maize embryo cDNA library by yeast one-hybrid screen using the ABRE2 sequence of the maize Cat1 gene as bait.  This protein, designated, ABRE binding protein 2 (ABP2), belongs to the bZIP transcription factor family.  Endogenous expression of ABP2 in maize can be detected in different tissues at various development stages, and can be induced by drought, salt, reactive oxygen species (ROS)-generating agents, and ABA treatment.  Constitutive expression of ABP2 in transgenic Arabidopsis plants enhanced tolerance to drought and salt stress, and increased sensitivity to ABA.  In exploring the mechanism by which ABP2 can stimulate abiotic stress tolerance, we found that ROS levels were reduced and expression of stress-responsive and carbon metabolism-related genes was enhanced by constitutive ABP2 expression in transgenic plants.  In short, we identified a maize bZIP transcription factor which can enhance both drought and salt tolerance of plants.
 
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Beneficial effects of silicon on photosynthesis of tomato seedlings under water stress
ZHANG Yi, SHI Yu, GONG Hai-jun, ZHAO Hai-liang, LI Huan-li, HU Yan-hong, WANG Yi-chao
2018, 17 (10): 2151-2159.   DOI: 10.1016/S2095-3119(18)62038-6
Abstract694)      PDF (930KB)(1005)      
Silicon can improve drought tolerance of plants, but the mechanism still remains unclear.  Previous studies have mainly concentrated on silicon-accumulating plants, whereas less work has been conducted in silicon-excluding plants, such as tomato (Solanum lycopersicum L.).  In this study, we investigated the effects of exogenous silicon (2.5 mmol L–1) on the chlorophyll fluorescence and expression of photosynthesis-related genes in tomato seedlings (Zhongza 9) under water stress induced by 10% (w/v) polyethylene glycol (PEG-6000).  The results showed that under water stress, the growth of shoot and root was inhibited, and the chlorophyll and carotenoid concentrations were decreased, while silicon addition improved the plant growth and increased the concentrations of chlorophyll and carotenoid.  Under water sterss, chlorophyll fluorescence parameters such as PSII maximum photochemical efficiency (Fv/Fm), effective quantum efficiency, actual photochemical quantum efficiency (ФPSII), photosynthetic electron transport rate (ETR), and photochemical quenching coefficient (qP) were decreased; while these changes were reversed in the presence of added silicon.  The expressions of some photosynthesis-related genes including PetE, PetF, PsbP, PsbQ, PsbW, and Psb28 were down-regulated under water stress, and exogenous Si could partially up-regulate their expressions.  These results suggest that silicon plays a role in the alleviation of water stress by modulating some photosynthesis-related genes and regulating the photochemical process, and thus promoting photosynthesis.
 
 
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Identification and characterization of Pichia membranifaciens Hmp-1 isolated from spoilage blackberry wine
WANG Ying, ZHAO Yan-cun, FAN Lin-lin, XIA Xiu-dong, LI Ya-hui, ZHOU Jian-zhong
2018, 17 (09): 2126-2136.   DOI: 10.1016/S2095-3119(18)62027-1
Abstract416)      PDF in ScienceDirect      
The pellicle-forming yeast could cause the quality deterioration of wine.  In this study, a pellicle-forming strain Hmp-1 was isolated from the spoilage blackberry wine, and identified as Pichia membranifaciens based on the morphology and partial nucleotide sequence of 26S rDNA.  The effects of fermentation conditions (ethanol, sulfur dioxide, sugar, and temperature) on the growth of P. membranifaciens strain Hmp-1 and Saccharomyces cerevisiae strain FM-S-115 (a strain used for the blackberry wine fermentation) were investigated, respectively.  The results indicated that Hmp-1 had lower resistance to these factors compared to FM-S-115, and the growth of Hmp-1 was completely inhibited by 10% (v/v) or 50 mg L–1 SO2 during the fermentation of blackberry wine.  These results suggested that Hmp-1 could effectively be controlled by increasing ethanol or SO2 concentration during the fermentation and storage of blackberry wine.  Furthermore, the analysis based on gas chromatography-mass spectrometry (GC-MS) showed that Hmp-1 remarkably decreased kinds of volatile compounds in blackberry wine, especially aldehydes and esters.  In addition, some poisonous compounds were detected in the blackberry wine fermented by FM-S-115 and Hmp-1.  These results suggested that Hmp-1 was a major cause leading to the quality deterioration of blackberry wine.
 
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Insecticide resistance of the field populations of oriental armyworm, Mythimna separata (Walker) in Shaanxi and Shanxi provinces of China
ZHAO Yu-yu, SU Li, LI Shuai, LI Yi-ping, XU Xiang-li, CHENG Wei-ning, WANG Yi, WU Jun-xiang
2018, 17 (07): 1556-1562.   DOI: 10.1016/S2095-3119(17)61787-8
Abstract518)      PDF in ScienceDirect      
Resistance of five field populations of Mythimna separata (Walker) collected from Shaanxi and Shanxi provinces of China to six different insecticides was evaluated by leaf dip method in the laboratory.  The results showed that all populations were relatively sensitive to emamectin benzoate with a resistance ratio (RR) of 0.583–1.583 folds.  All populations showed susceptible or low level resistance to chlorantraniliprole and beta-cypermethrin.  Compared with a relatively susceptible strain of M. separata, the resistance level of the whole populations ranged from susceptible to moderate to chlorpyrifos and lambda-cyhalothrin, moderate to high to phoxim (RR=19.367–70.100) except for population from Sanyuan County (RR=2.567).  Pair-wise correlation analysis among different insecticides indicated that chlorpyrifos has a significantly positive and significant correlation with emamectin benzoate.  Chlorantraniliprole didn’t have significant correlation with emamectin benzoate, chlorpyrifos and phoxim.  Therefore, emamectin benzoate, chlorantraniliprole and beta-cypermethrin are recommended to control oriental armyworm.  Meanwhile, to postpone the occurrence and development of insecticide resistance in Shaanxi and Shanxi provinces, alternative and rotational application of insecticides between chlorantraniliprole and emamectin benzoate or chlorpyrifos is necessary.
 
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A wheat gene TaSAP17-D encoding an AN1/AN1 zinc finger protein improves salt stress tolerance in transgenic Arabidopsis
XU Qiao-fang, MAO Xin-guo, WANG Yi-xue, WANG Jing-yi, XI Ya-jun, JING Rui-lian
2018, 17 (03): 507-516.   DOI: 10.1016/S2095-3119(17)61681-2
Abstract687)      PDF in ScienceDirect      
The stress-associated protein (SAP) multigene family is conserved in both animals and plants.  Its function in some animals and plants are known, but it is yet to be deciphered in wheat (Triticum aestivum L.).  We identified the wheat gene TaSAP17-D, a member of the SAP gene family with an AN1/AN1 conserved domain.  Subcellular localization indicated that TaSAP17-D localized to the nucleus, cytoplasm, and cell membrane.  Expression pattern analyses revealed that TaSAP17-D was highly expressed in seedlings and was involved in NaCl response, polyethylene glycol (PEG), cold, and exogenous abscisic acid (ABA).  Constitutive expression of TaSAP17-D in transgenic Arabidopsis resulted in enhanced tolerance to salt stress, confirmed by improved multiple physiological indices and significantly upregulated marker genes related to salt stress response.  Our results suggest that TaSAP17-D is a candidate gene that can be used to protect crop plants from salt stress.  
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The effect of dehydrogenase enzyme activity in glycolysis on the colour stability of mutton during postmortem storage
XIN Jian-zeng, LI Zheng, LI Xin, LI Meng, WANG Ying, YANG Fu-min, ZHANG De-quan
2017, 16 (11): 2646-2654.   DOI: 10.1016/S2095-3119(16)61622-2
Abstract623)      PDF in ScienceDirect      
This study investigated the influence of activities of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and lactate dehydrogenase-B (LDH-B) on the colour stability of mutton.  From 60 sheep (Bayannur mutton sheep), 15 longissimus dorsi (LD) muscles were selected on the basis of colour stability (R630/580 and a* value) during the storage and classified into three groups (5 for each group) as high colour stability (HCS), intermediate colour stability (ICS) and low colour stability (LCS).  The activities of GAPDH and LDH-B, muscle colour attributes, nicotinamide adenine dinucleuotide (NADH) concentration and lactate concentration were measured.  The samples in HCS had higher activities of GAPDH and LDH-B than the samples in the LCS, and the samples in the HCS group also possessed higher NADH and lower lactate concentration.  The higher activity of dehydrogenase enzyme may result in higher NADH concentrations and colour stability in muscle tissue.  The results suggest that the activity of GAPDH and LDH-B may also play a role in maintaining colour stability.
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Responses of N2O reductase gene (nosZ)-denitrifier communities to long-term fertilization follow a depth pattern in calcareous purplish paddy soil
WANG Ying-yan, LU Sheng-e, XIANG Quan-ju, YU Xiu-mei, ZHAO Ke, ZHANG Xiao-ping, TU Shihua, GU Yun-fu
2017, 16 (11): 2597-2611.   DOI: 10.1016/S2095-3119(17)61707-6
Abstract1024)      PDF in ScienceDirect      
    The effect of long-term fertilization on soil denitrifying communities was analysed by measuring the abundance and diversity of the nitrous oxide (N2O) reductase gene, nosZ.  Soil samples were collected from plots of a long-term fertilization experiment established in 1982 in Suining City, China.  The fertilizer treatments were no fertilizer (CK), three chemical fertilizer (CF) treatments (N, NP, NPK), manure (M) alone, and manure with chemical fertilizers (NM, NPM, NPKM).  The abundance and diversity of the denitrifying bacteria were assessed by real-time quantitative PCR, terminal restriction fragment length polymorphism (T-RFLP), and cloning and sequencing of nosZ genes.  The diversity and abundance of nosZ-denitrifiers was higher in soil amended with manure and chemical fertilizers (CFM) than in soil amended with CF alone, and the highest in topsoil (0–20 cm).  The nosZ-denitrifier community composition was more complex in CFM soil than in CF soil.  Specific species were detected only in the CFM soil.  The abundance of nosZ-denitrifier in the NPKM treatment was approximately two times higher than that in the CK, N, and NPK treatments.  Most of the cloned nosZ sequences were closely related to nosZ sequences from Bradyrhizobiaceae and Rhodospirillaceae in Alphaproteobacteria.  Of the measured abiotic factors, soil organic matter correlated significantly with the abundance (P<0.01); available phosphorus correlated significantly with the topsoil community composition (P<0.01), whereas soil organic matter correlated significantly with the subsoil (20–90 cm) community composition (P<0.01). This study demonstrated that long-term CFM fertilization affected both the abundance and composition of the nosZ-denitrifier community. 
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