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Light intensity regulates the sexual behaviors of oriental fruit fly Bactrocera dorsalis under laboratory conditions
REN Cong, ZHANG Jie, YUAN Jin-xi, WU Yun-qi-qi-ge, YAN Shan-chun, LIU Wei, WANG Gui-rong
2023, 22 (9): 2772-2782.   DOI: 10.1016/j.jia.2023.04.025
Abstract193)      PDF in ScienceDirect      

The oriental fruit fly, Bactrocera dorsalis (Hendel), is a devastating pest of citrus fruits.  After successful mating, adult females insert their eggs into the ripened fruit, resulting in moldy and rotten fruit and causing great economic losses for the citrus industry.  In the field, flies initiate copulatory behaviors as twilight approaches, and decreasing light intensity in this period is the normal stimulus for copulation.  In this study, ten light intensities ranging from 0–30 000 lux were set to identify the typical intensity that strongly regulates the copulation behavior of Bdorsalis.  Three light intensities found to regulate the copulation behavior were then selected to verify their effects on adult male wing fanning and female chemotaxis towards 2,3,5-trimethylpyrazine (TMP).  At last, strong light and complete darkness were artificially combined in the lab to verify whether they could prevent copulation to inform behavioral manipulation of oriental flies in the future.  The results indicated that adult flies generally initiated copulatory behaviors at low light intensity (<1 000 lux).  
Stronger light significantly prevented copulation in proportion to intensity, with nearly no copulation events initiated when light intensity was above 20 000 lux.  Both male wing fanning and female chemotaxis towards TMP were attenuated as light intensity became stronger.  However, at 10 000 lux, males still fanned their wings to a certain extent while TMP completely lost its attractiveness to females.  In the darkness, adults did not initiate any sexual behaviors, e.g., copulation, wing fanning, or chemotaxis to TMP.  One hour of strong light (10 000 lux) combined with continuous darkness completely prevented mating.  These results show that light condition is an essential factor for copulatory behaviors in the oriental fruit fly.  Researchers could thus manipulate light conditions artificially or disrupt the molecular target in flies’ light transduction pathway to develop environmentally-friendly techniques to control this pest.

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Development and characterization of wheat–Aegilops kotschyi 1Uk(1A) substitution line with positive dough quality parameters

JIANG Yun, WANG De-li, HAO Ming, ZHANG Jie, LIU Deng-cai
2023, 22 (4): 999-1008.   DOI: 10.1016/j.jia.2022.08.020
Abstract239)      PDF in ScienceDirect      

Exploring novel high molecular weight glutenin subunits (HMW-GSs) from wild related species is a strategy to improve wheat processing quality.  The objective of the present investigation was to identify the chromosomes of the wheat-alien introgression line N124, derived from the hybridization between Triticum aestivum with Aegilops kotschyi, and characterize the effects on quality-related traits.  Fluorescence in situ hybridization karyotypes showed that N124 is a disomic 1Uk(1A) substitution line.  Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and reversed-phase high-performance liquid chromatography verified N124 expressed two HMW-GSs of the Ae. kotschyi parent.  PacBio RNA sequencing and phylogenetic analysis confirmed that the two HMW-GSs were Ukx and Uky.  Compared to the wheat parent, the substitution line had no obvious agronomic defects except fewer grains per spike but improved several major quality parameters.  It can be served as a donor or bridge material for wheat quality improvement.

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Mitochondrial dynamics caused by QoIs and SDHIs fungicides depended on FgDnm1 in Fusarium graminearum
KANG Jin-bo, ZHANG Jie, LIU Yin-kai, SONG Ji-chang, OU Jian-lin, TAO Xian, ZHOU Ming-guo, DUAN Ya-bing
2023, 22 (2): 481-494.   DOI: 10.1016/j.jia.2022.08.118
Abstract436)      PDF in ScienceDirect      

Fusarium head blight (FHB) caused by Fusarium graminearum is a devastating fungal disease on small grain cereal crops, because it reduces yield and quality and causes the mycotoxin contamination to the grain.  Dynamins and dynamin-related proteins (DRPs) are large GTPase superfamily members, which are typically involved in the budding and division of vesicles in eukaryotic cells, but their roles in Fusarium spp. remain unexplored.  Here, we found that FgDnm1, a DRP and homolog to Dnm1 in Saccharomyces cerevisiae, contributes to the normal fungal growth, sexual reproduction and sensitivity to fungicides.  In addition, we found FgDnm1 co-localizes with mitochondria and is involved in toxisome formation and deoxynivalenol (DON) production.  Several quinone outside inhibitors (QoIs) and succinate dehydrogenase inhibitors (SDHIs) cause fragmentated morphology of mitochondria.  Importantly, the deletion of FgDnm1 displays filamentous mitochondria and blocks the mitochondrial fragmentation induced by QoIs and SDHIs.  Taken together, our studies uncover the effect of mitochondrial dynamics in fungal normal growth and how such events link to fungicides sensitivity and toxisome formation.  Thus, we concluded that altered mitochondrial morphology induced by QoIs and SDHIs depends on FgDnm1.

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BnaSD.C3 is a novel major quantitative trait locus affecting semi-dwarf architecture in Brassica napus L.
WANG Xiao-dong, CAI Ying, PANG Cheng-ke, ZHAO Xiao-zhen, SHI Rui, LIU Hong-fang, CHEN Feng, ZHANG Wei, FU San-xiong, HU Mao-long, HUA Wei, ZHENG Ming, ZHANG Jie-fu
2023, 22 (10): 2981-2992.   DOI: 10.1016/j.jia.2023.02.017
Abstract217)      PDF in ScienceDirect      

Plant height is a key plant architectural trait that affects the seed yield, harvest index and lodging resistance in Brassica napus L., although the genetic mechanisms affecting plant height remain unclear.  Here, a semi-dwarf mutant, df34, was obtained by ethyl methanesulphonate-induced mutagenesis.  Genetic analysis showed that the semi-dwarf phenotype is controlled by one semi-dominant gene, which was located on chromosome C03 using a bulked segregant analysis coupled with whole-genome sequencing, and this gene was named BnaSD.C3.  Then BnaSD.C3 was fine-mapped to a 297.35-kb segment of the “Darmor-bzh” genome, but there was no potential candidate gene for the semi-dwarf trait underlying this interval.  Furthermore, the interval was aligned to the Zhongshuang 11 reference genome.  Finally, combining structural variation analysis, transcriptome sequencing, phytohormone analyses and gene annotation information, BnaC03G0466900ZS and BnaC03G0478900ZS were determined to be the most likely candidate genes affecting the plant height of df34.  This study provides a novel major locus for breeding and new insights into the genetic architecture of plant height in Bnapus

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The transcription factor FgNsf1 regulates fungal development, virulence and stress responses in Fusarium graminearum
SHI Dong-ya, REN Wei-chao, WANG Jin, ZHANG Jie, Jane Ifunanya MBADIANYA, MAO Xue-wei, CHEN Chang-jun
2021, 20 (8): 2156-2169.   DOI: 10.1016/S2095-3119(20)63339-1
Abstract154)      PDF in ScienceDirect      
Nutrient and stress factor 1 (Nsf1), a transcription factor containing the classical Cys2-His2 (C2H2) zinc finger motif, is expressed under non-fermentable carbon conditions and in response to salt stress in Saccharomyces cerevisiae.  However, the role of Nsf1 in filamentous fungi is not well understood.  In this study, the orthologue of Nsf1 was investigated in Fusarium graminearum (named FgNsf1), a causal agent of Fusarium head blight (FHB).  The functions of FgNsf1 were evaluated by constructing a FgNSF1 deletion mutant, designated as ΔFgNsf1, and its functional complementation mutant ΔFgNsf1-C.  Gene deletion experiments showed that the mycelial growth rate, asexual and sexual reproduction of ΔFgNsf1 were significantly reduced, but the pigment production of ΔFgNsf1 was remarkably increased compared with the PH-1 and ΔFgNsf1-C.  In addition, the tolerance of ΔFgNsf1 to osmotic pressures, cell wall-damaging agents and oxidative stress increased significantly.  Sensitivity tests to different fungicides revealed that ΔFgNsf1 exhibited increased sensitivity to carbendazim (MBC) and tebuconazole, and enhanced tolerance to fludioxonil and iprodione than PH-1 and ΔFgNsf1-C.  The virulence of ΔFgNsf1 to wheat coleoptiles and flowering wheat heads were dramatically decreased, which was consistent with the decrease in the yield of deoxynivalenol (DON).  All of these defects were restored by target gene complementation.  These results indicated that FgNsf1 plays a crucial role in vegetative growth, asexual and sexual reproduction, stress responses, fungicide sensitivity, and full virulence in F. graminearum.
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Genetic parameter estimation and genome-wide association study (GWAS) of red blood cell count at three stages in a Duroc×Erhualian pig population
NAN Jiu-hong, YIN Li-lin, TANG Zhen-shuang, CHEN Jian-hai, ZHANG Jie, WANG Hai-yan, DU Xiao-yong, LIU Xiang-dong
2020, 19 (3): 793-799.   DOI: 10.1016/S2095-3119(19)62773-5
Abstract135)      PDF in ScienceDirect      
Red blood cells play an essential role in the immune system.  Moreover, red blood cell count (RBC) is an important clinical indicator of various diseases, including anemia, type 2 diabetes and the metabolic syndrome.  Thus, it is necessary to reveal the genetic mechanism of RBC for animal disease resistance breeding.  However, quite a few studies had focused on porcine RBC, especially at different stages.  Thus, studies on porcine RBC at different stages are needed for disease resistant breeding.  In this study, the porcine RBC of 20-, 33-, and 80-day old were measured, and genetic parameter estimation and genome-wide association study (GWAS) were both performed.  As a result, the heritability was about 0.6 at the early stages, much higher than that at 80 days.  Nine novel genome wide significant single nucleotide polymorphisms (SNPs), located at Sus scrofa chromosome (SSC)3, 4, 8, 9, 10 and 15, respectively, were identified.  Further, TGFβ2, TMCC2 and PPP1R15B genes were identified as important candidate genes of porcine red blood cell count.  So different SNPs and candidate genes were found significantly associated with porcine RBC at different stages, suggesting that different genes might play key roles on porcine RBC at different stages.  Overall, new evidences were offered in this study for the genetic bases of animal RBC, and that the SNPs and candidate genes would be useful for disease resistant breeding of pig.
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One-time fertilization at first flowering improves lint yield and dry matter partitioning in late planted short-season cotton
LUO Hong-hai, WANG Qiang, ZHANG Jie-kun, WANG Lei-shan, LI Ya-bing, YANG Guo-zheng
2020, 19 (2): 509-517.   DOI: 10.1016/S2095-3119(19)62623-7
Abstract132)      PDF in ScienceDirect      
Cotton producers have substantially reduced their inputs (labor, nutrients, and management) mainly by adopting a short-season cropping management that is characterized by late sowing, high density, and reduced fertilization with one-time application at the first bloom stage without lint yield reduction.  However, it has been hypothesized that one-time fertilization at an earlier growth stage could be a more effective and economic management practice.  A two-year field experiment was conducted by applying five fertilizer one-time fertilization at 0 (FT1), 5 (FT2), 10 (FT3), 15 (FT4), and 20 (FT5) days after the first flower appeared in the field and one three-split fertilizer application taken as the conventional control (FT6), making six treatments altogether.  Cotton growth period, biomass accumulation, yield, and its formation were quantified.  The results showed that the one-time fertilization did not affect the cotton growth progress as compared to FT6, however, the total crop cycles for FT3–FT5 were 3 days shorter.  FT1 produced the highest cotton lint yield (1 396 kg ha–1), which was similar to the FT6 but higher than the other treatments, and could be attributed to more bolls per unit area and higher lint percentage. Cotton yield was positively correlated with cotton plant biomass accumulated.  FT1 had both the highest average (VT) (193.7 kg ha–1 d–1) and the highest maximum (VM) (220.9 kg ha–1 d–1) rates during the fast biomass accumulation period.  These results suggest that one-time fertilizer application at the first flower stage might be an adjustment that is more effective than at first bloom, and allowed for easier decision making for application date due to non counting of plants with flowers is needed.
 
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Alanine-substituted mutant on Gly373 and Asn375 of Cry1Ai-h-loop 2 causes reduction in both toxicity and binding against Helicoverpa armigera
LIU Yu-xiao, ZHOU Zi-shan, LIANG Ge-mei, SONG Fu-ping, ZHANG Jie
2019, 18 (5): 1064-1071.   DOI: 10.1016/S2095-3119(18)61933-1
Abstract165)      PDF in ScienceDirect      
Cry1Ai-h-loop 2 is a mutant of Cry1Ai constructed by exchanging loop 2 from Cry1Ah protein and shows insecticidal activity against Helicoverpa armigera.  The toxicity of Cry1Ai-h-loop 2, in contrast to the very low toxicity of Cry1Ai, is closely associated with the eleven residues in the loop 2 region.  To characterize the key sites of loop 2 in Cry1Ai-h-loop 2, alanine-substituted mutants were generated.  The toxicity of these mutants against H. armigera indicated that dual-mutant on Gly373 and Asn375 caused a significant decrease in toxic activity.  ELISA binding and competition binding assays demonstrated that the reduction of toxicity in the mutant of interest was correlated with decreased binding affinity.
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Transcriptional profile of gene clusters involved in the methylerythritol phosphate pathway in Bacillus subtilis 916
XIAO Ya-jing, GAO Tan-tan, PENG Qi, ZHANG Jie, SUN Dong-mei, SONG Fu-ping
2019, 18 (3): 644-655.   DOI: 10.1016/S2095-3119(18)62001-5
Abstract206)      PDF (680KB)(162)      
The methylerythritol phosphate pathway is responsible for the biosynthesis of terpenoids, the largest class of secondary metabolites.  Although the structures and functions of the proteins involved in this pathway have been well studied in Bacillus subtilis, only a few studies have reported the transcriptional profile of the genes involved.  Therefore, we analyzed methylerythritol phosphate pathway genes in the genome of B. subtilis 916, which has been developed as a biological control agent against some rice diseases in China.  Our results showed that methylerythritol phosphate pathway genes were distributed throughout the genome of this strain.  These genes were transcribed during both the exponential and stationary phases.  We further confirmed the transcription units of dxs, dxr, ispD, ispF, ipK, ispG, ispH, idi, and ispA in B. subtilis 916 through reverse transcription-PCR analyses; the results showed that these nine genes were located in seven different operons.  The transcript start sites of the seven different operons were determined by 5´-rapid amplification of cDNA ends-PCR.  Thus, our study provides a molecular basis at the transcriptional level for investigating homoterpene synthesis in the methylerythritol phosphate pathway of B. subtilis 916.
 
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Inheritance and molecular characterization of resistance to AHAS-inhibiting herbicides in rapeseed
HU Mao-long, PU Hui-ming, GAO Jian-qin, LONG Wei-hua, CHEN Feng, ZHOU Xiao-ying, ZHANG Wei, PENG Qi, CHEN Song, ZHANG Jie-fu
2017, 16 (11): 2421-2433.   DOI: 10.1016/S2095-3119(17)61659-9
Abstract791)      PDF in ScienceDirect      
Rapeseed is a very important oil crop in China; however, its production is challenging due to the absence of effective weed management strategies.  This is predominantly because of a shortage of herbicide resistance genes.  Acetohydroxyacid synthase (AHAS) herbicides inhibit AHAS, a key enzyme involved in branched-chain amino acid synthesis that is required for plant growth.  A rapeseed line designated M342 with AHAS herbicide resistance was developed through seed mutagenesis and was studied to assess the level and mode of inheritance of the resistance and to identify the molecular basis of resistance.  M342 possessed a high level of cross-resistance to sulfonylureas (SUs) and imidazolinones (IMIs).  This resistance was due to AHAS insensitivity to these herbicides and was inherited as a dominant trait conferred by a single nuclear-encoded gene.  Molecular analysis revealed the presence of a Trp574Leu mutation in M342, and an allele-specific cleaved amplified polymorphic sequence (AS-CAPS) marker was developed and cosegregated with herbicide resistance in the F2, BC1, and BC2 populations.  This mutation altered the transcript levels of BnAHAS1 and BnAHAS3 in M342 compared with those in the wild type, but it did not affect the agronomic or quality traits.  The simple genetic inheritance of this mutation and the availability of the cleaved amplified polymorphic sequence (CAPS) marker and herbicide resistance gene should facilitate the development of herbicide-resistant rapeseed cultivars for effective weed control in China.  
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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
Abstract953)      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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Identification of similar transcriptional regulatory mechanisms in multiple cry genes in Bacillus thuringiensis HD12
SONG Zhi-ru, PENG Qi, SHU Chang-long, ZHANG Jie, SUN Dong-mei, SONG Fu-ping
2017, 16 (01): 135-143.   DOI: 10.1016/S2095-3119(16)61398-9
Abstract690)           
Bacillus thuringiensis subspecies morrisoni strain HD12, whose genome harbors multiple insecticidal protein-encoding genes, includes eight cry genes, as indicated by genome sequencing.  This strain produces crystals that are toxic to lepidopteran species.  These crystal inclusions were purified by sucrose gradients and sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by liquid chromatography-mass spectrometry, and found to contain five proteins (Cry1Da, Cry1Ae, Cry1Bb, Cry1Fb, and Cry1Ja).  The transcriptional activities of the cry1Da, cry1Ae, cry1Bb, cry1Fb, and cry1Ja promoters indicated that transcription of cry1Da is controlled by SigE; however, the other four cry genes were found to be controlled by both SigE and SigK.  The activities of the cry1Ja and cry1Fb promoters were the strongest among the five genes studied.  These promoters were therefore used to direct the expression of the Cry1Ac- and Cry2Ab-encoding genes concurrently in a single strain.  Our findings indicate that promoters with the same transcriptional profile may be used to direct the expression of different cry genes in one strain.  Our results are expected to be valuable for the construction of strains with efficient expression of multiple cry genes in order to overcome current limitations associated with the application of B. thuringiensis-based insecticides.
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Separation and purification of deoxynivalenol (DON) mycotoxin from wheat culture using a simple two-step silica gel column chromatography
ZHAO Xiu-mei, LI Rong-jia, ZHOU Chuang, ZHANG Jie, HE Cheng-hua, ZHENG Ya-ting, WU Wen-da
2016, 15 (3): 694-701.   DOI: 10.1016/S2095-3119(15)61098-X
Abstract1871)      PDF in ScienceDirect      
Deoxynivalenol (DON) is a type B trichothecenes mycotoxin produced by several Fusarium species, often found in foodstuffs for humans and animals. DON is in great demand for the toxicological researches both in vivo and in vitro. In this work, wheat culture was inoculated with a Fusarium graminearum PH-1 strain for DON production. The solvent system for crude extraction was acetonitrile-water (84:16, v/v). A simple two-step silica gel column chromatography was employed to separate the DON mycotoxin from wheat culture, combined with preparative high performance liquid chromatography (preparative HPLC) to purify the compound. The solvent system for the second silica gel column chromatography was methylene chloride-methanol (17:1, v/v), which provided a good elution effect selected on thin layer chromatography (TLC). The target compound was identified by HPLC, and the chemical structure was confirmed by mass spectrometry (MS) and 1H and 13C nuclear magnetic resonance (NMR) spectroscopy. A total of 433 mg of purified DON was obtained from 1 kg of wheat culture, with a purity of 99.01%. The study had provided an easy-operating and cost-effective method to isolate an expensive compound in a simple way.
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Comparison and optimization of the method for Cry1Ac protoxin preparation in HD73 strain
ZHOU Zi-shan, YANG Su-juan, SHU Chang-long, SONG Fu-ping, ZHOU Xue-ping, ZHANG Jie
2015, 14 (8): 1598-1603.   DOI: 10.1016/S2095-3119(14)60950-3
Abstract2095)      PDF in ScienceDirect      
Bacillus thuringiensis is one of the most widely used bioinsecticides, and cry gene is the major insecticidal gene. Because Cry1Ac protein shows strong toxicity against many lepidopteran species, it has been applied widely in spraying products and transgenic Bt-crops. The preparation of Cry protoxin is the first step in the very important processes of understanding the insecticidal mechanism, resistance screening, and biosafety assessments. The media for crystal production and the method for Cry protoxin preparation were varied, however, it was not clear which was better for preparing a larger amount of Cry protoxin. In this paper, three media for crystal production and the method for Cry1Ac protoxin preparation from HD73 strain were compared to find an efficacious way to prepare a large number of Cry1Ac protoxin. The results showed that the 1/2 LB (Luria-Bertani) medium was the ideal medium for crystal production, because the total yield of Cry1Ac protoxin in 300 mL 1/2 LB medium was (112.38±5.64) mg, the highest one among three media; the repeated crystal solubilization method was better for the preparation of the Cry protoxin comparing with the continuous crystal solubilization method. It will be a reference for other Cry protoxin preparation, especially for larger number.
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Developing transgenic maize (Zea mays L.) with insect resistance and glyphosate tolerance by fusion gene transformation
SUN He, LANG Zhi-hong, LU Wei, ZHANG Jie, HE Kang-lai , ZHU Li, LIN Min, HUANG Da-fang
2015, 14 (2): 305-313.   DOI: 10.1016/S2095-3119(14)60855-8
Abstract1803)      PDF in ScienceDirect      
Using linker peptide LP4/2A for multiple gene transformation is considered to be an effective method to stack or pyramid several traits in plants. Bacillus thuringiensis (Bt) cry gene and epsps (5-enolpyruvylshikimate-3-phosphate synthase) gene are two important genes for culturing pest-resistant and glyphosate-tolerant crops. We used linker peptide LP4/2A to connect the Bt cry1Ah gene with the 2mG2-epsps gene and combined the wide-used manA gene as a selective marker to construct one coordinated expression vector called p2EPUHLAGN. The expression vector was transferred into maize by Agrobacterium tumefaciens-mediated transformation, and 60 plants were obtained, 40% of which were positive transformants. Molecular detection demonstrated that the two genes in the fusion vector were expressed simultaneously and spliced correctly in translation processing; meanwhile bioassay detection proved the transgenic maize had preferable pest resistance and glyphosate tolerance. Therefore, linker peptide LP4/2A provided a simple and reliable strategy for producing gene stacking in maize and the result showed that the fusion gene transformation system of LP4/2A was feasible in monocot plants.
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The Minimal Active Fragment of the Cry1Ai Toxin is Located Between 36I and 605I
ZHOU Zi-shan, LIN Hui-yan, LI Ying, SHU Chang-long, SONG Fu-ping , ZHANG Jie
2014, 13 (5): 1036-1042.   DOI: 10.1016/S2095-3119(13)60532-8
Abstract1894)      PDF in ScienceDirect      
The novel cry1Ai gene that cloned from Bacillus thuringiensis strain SC6H8 encoded a protein exhibiting strong toxicity against Plutella xylostella and Chilo suppressalis in our previous study. Using the available information for the active fragments of other Cry toxins, eight truncated fragments were constructed to identify the minimal active fragment of Cry1Ai. All truncated fragments were expressed in Escherichia coli strain BL21 (DE3), and the insecticidal activity against 2nd- instar P. xylostella larvae was assessed using full-length Cry1Ai as a positive control. The results indicate that the minimal active fragment of the Cry1Ai toxin against P. xylostella is located between amino acid residues 36I and 605I, which is smaller than the regions previously reported for Cry1A. The first two amino acids (34T and 35P) on helix α-1 and whole helix α-2 of domain I and sheet β-32 of domain III are necessary for Cry1Ai toxin to keep its toxicity against P. xylostella.
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Acquisition of Insect-Resistant Transgenic Maize Harboring a Truncated cry1Ah Gene via Agrobacterium-Mediated Transformation
LI Xiu-ying, LANG Zhi-hong, ZHANG Jie, HE Kang-lai, ZHU Li , HUANG Da-fang
2014, 13 (5): 937-944.   DOI: 10.1016/S2095-3119(13)60531-6
Abstract2075)      PDF in ScienceDirect      
A novel insecticidal gene cry1Ah was cloned from Bacillus thuringiensis isolate BT8 previously for plant genetic engineering improvement. Truncated active Cry1Ah toxin has a toxicity level similar to that of the full-length Cry1Ah toxin. In this study, plant expression vector pMhGM harboring truncated cry1Ah gene was transformed into maize (Zea mays L.) immature embryos by Agrobacterium tumefaciens-mediated transformation at which maize alcohol dehydrogenase matrix attachment regions (madMARs) were incorporated on both sides of the gene expression cassette to improve gene expression. A total of 23 PCR positive events were obtained with a transformation efficiency of 5% around. Bioassay results showed that events 1-4 and 1-5 exhibited enhanced resistance to the Asian corn borer (Ostrinia furnacalis). These two events were further confirmed by molecular analysis. Southern blot suggested that a single copy of the cry1Ah gene was successfully integrated into the maize genome. Western blot and ELISA showed that the foreign gene cry1Ah was expressed stably at high level in maize and could be inherited stably over generations. The results of a bioassay of T1-T4 transgenic maize plants indicated that the transgenic plants were highly toxic to the Asian corn borer and their resistance could be inherited stably from generation to generation. Thus, events 1-4 and 1-5 are good candidates for the breeding of insect-resistant maize.
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The Influence of Bt-Transgenic Maize Pollen on the Bacterial Diversity in the Midgut of Chinese Honeybees, Apis cerana cerana
JIANG Wei-yu, GENG Li-li, DAI Ping-li, LANG Zhi-hong, SHU Chang-long, LIN Yi, ZHOU Ting, SONG Fu-ping , ZHANG Jie
2013, 12 (3): 474-482.   DOI: 10.1016/S2095-3119(13)60248-8
Abstract1377)      PDF in ScienceDirect      
Using culture-independent technique polymerase chain reaction-denaturing gradient gel electrophoresis (PCR-DGGE) and conventional culture techniques, ecological risk of transgenic maize pollen on gut bacteria of the Chinese honeybee, Apis cerana cerana, was assessed. Honeybees were fed with Bt-transgenic maize pollen, non-transgenic near isoline pollen, linear cry1Ah gene (800 ng mL-1) and supercoiled plasmid DNA (800 ng mL-1) under laboratory conditions. The DGGE profile showed that the number of DGGE bands varied from 10.7 to 14.7 per sample, and the Shannon’s index ranged from 0.85 to 1.00. The similarity calculated by PAST was mostly above 92%, indicating no obvious changes among treatments or within replicates. 14 bacterial strains affiliated with Alphaproteobacteria, Gammaproteobacteria, Firmicutes, and Actinobacteria were isolated and characterized on media under aerobic and anaerobic conditions. These results demonstrated that transgenic cry1Ah maize pollen did not induce significant changes of the honeybee gut bacterial community composition under laboratory conditions.
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Application of Mulching Materials of Rainfall Harvesting System for Improving Soil Water and Corn Growth in Northwest of China
HAN Juan, JIA Zhi-kuan, HAN Qing-fang , ZHANG Jie
2013, 12 (10): 1712-1721.   DOI: 10.1016/S2095-3119(13)60342-1
Abstract1912)      PDF in ScienceDirect      
The ridge and furrow rainfall harvesting (RFRH) system is used for dryland crop production in northwest of China. To determine the effects of RFRH using different mulching materials on corn growth and water use efficiency (WUE), a field experiment was conducted during 2008-2010 at the Heyang Dryland Experimental Station, China. Four treatments were used in the study. Furrows received uncovered mulching in all RFRH treatments whereas ridges were mulched with plastic film (PF), biodegradable film (BF) or liquid film (LF). A conventional flat field without mulching was used as the control (CK). The results indicated that the average soil water storage at depths of 0-200 cm were 8.2 and 7.3%, respectively higher with PF and BF than with CK. However, LF improved soil water storage during the early growth stage of the crop. Compared with CK, the corn yields with PF and BF were increased by 20.4 and 19.4%, respectively, and WUE with each treatment increased by 23.3 and 21.7%, respectively. There were no significant differences in corn yield or WUE with the PF and BF treatments. The net income was the highest with PF, followed by BF, and the 3-yr average net incomes with these treatments were increased by 2 559 and 2 430 CNY ha-1, respectively, compared with CK. BF and PF had similar effects in enhancing the soil water content, crop yield and net income. Therefore, it can be concluded that biodegradable film may be a sustainable ecological alternative to plastic film for use in the RFRH system in northwest of China.
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The stress regulator FgWhi2 and phosphatase FgPsr1 play crucial roles in the regulation of secondary metabolite biosynthesis and the response to fungicides in Fusarium graminearum
Zhang Jie, Gao Han, Ren Fuhao, Zhou Zehua, Wu Huan, Zhao Huahua, Zhang Lu, Zhou Mingguo, Duan Yabing
DOI: 10.1016/j.jia.2024.01.003 Online: 19 December 2023
Abstract78)      PDF in ScienceDirect      

In yeast, the stress-responsive protein Whi2 interacts with phosphatase Psr1 to form a complex that regulates cell growth, reproduction, infection, and the stress response. However, the roles of Whi2 and Psr1 in Fusarium graminearum remain unclear. In this study, we identified homologous genes of WHI2 and PSR1 in F. graminearum and evaluated their functions by constructing deletion mutants. By comparing the responses of the mutants to different stressors, we found that FgWHI2 and FgPSR1 were involved in responding to osmotic, cell wall and cell membrane stresses, while also affecting the sexual and asexual reproduction in F. graminearum. Our studies demonstrated that FgWHI2 and FgPSR1 regulate the biosynthesis of ergosterol and the transcriptional level of FgCYP51C, which is a CYP51 paralogues unique to Fusarium species. This study also found that the deoxynivalenol (DON) production of FgWHI2 and FgPSR1 deletion mutants was reduced by ≥ 90% and DON production was positively correlated with the transcriptional levels of FgWHI2 and FgPSR1. In addition, we observed that FgWHI2 and FgPSR1 were involved in regulating the sensitivity of F. graminearum to chlorothalonil, fluazinam, azoxystrobin, phenamacril, and oligomycin. This study revealed the existence of cross-resistance between chlorothalonil and fluazinam. chlorothalonil and fluazinam inhibited DON biosynthesis by suppressing the expression of FgWHI2. Interestingly, the subcellular localization of FgWhi2 and FgPsr1 was significantly altered after treatment with chlorothalonil and fluazinam, with increased co-localization. Collectively, these findings indicate that FgWHI2 and FgPSR1 play crucial roles in stress response mechanisms, reproductive processes, secondary metabolite synthesis, and fungicide sensitivity in F. graminearum.

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